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1,000 | 15,632,662 | 1,612 | The invention relates to a lubricating member for a razor cartridge comprising a metathesized unsaturated polyol ester for improved lubrication. | 1. A lubricating member for use on a hair removal device, said lubricating member comprising a lubricating material comprising a) from 1% to 99% by weight of methathesized unsaturated polyol ester methathesized unsaturated polyol ester, having one or more of the following properties:
(i) a weight average molecular weight of from about 5,000 Daltons to about 50,000 Daltons; (ii) an oligomer index from greater than 0 to 1; and (iii) an iodine value of from about 30 to about 200. 2. A device according to claim 1, wherein said metathesized unsaturated polyol ester has a weight average molecular weight of from about 5,000 Daltons to about 50,000 Daltons. 3. A device according to claim 1 wherein said metathesized unsaturated polyol ester has an iodine value of from about 30 to about 200. 4. A lubricating member for use on a hair removal device, said lubricating member comprising a lubricating material comprising a) from 1% to 99% by weight of composition comprising:
a) a metathesized unsaturated polyol ester, said metathesized unsaturated polyol ester having a weight average molecular weight of from about 2,000 Daltons to about 50,000 Daltons; and one or more of the following properties:
(i) a free hydrocarbon content, based on total weight of metathesized unsaturated polyol ester of from about 0% to about 5%;
(ii) an oligomer index from greater than 0 to 1; and
(iii) an iodine value of from about 8 to about 200. 5. A lubricating member according to claim 4, wherein said metathesized unsaturated polyol ester has an iodine value of from about 10 to about 200. 6. A lubricating member according to claim 4, wherein said metathesized unsaturated polyol ester has an oligomer index from about 0.001 to 1. 7. A lubricating member according to claim 1, wherein said metathesized unsaturated polyol ester has a free hydrocarbon content, based on total weight of metathesized unsaturated polyol ester, of from about 0% to about 5%. 8. A lubricating member according to claim 1, said composition comprising, based on total composition weight, from about 0.1% to about 50% of said metathesized unsaturated polyol ester. 9. A lubricating member according to claim 1, wherein the metathesized unsaturated polyol ester is metathesized at least once. 10. A lubricating member according to claim 1, wherein said metathesized unsaturated polyol ester is derived from a natural polyol ester and/or a synthetic polyol ester, preferably said natural polyol ester is selected from the group consisting of a vegetable oil, an animal fat, an algae oil and mixtures thereof; and said synthetic polyol ester is derived from a material selected from the group consisting of ethylene glycol, propylene glycol, glycerol, polyglycerol, polyethylene glycol, polypropylene glycol, poly(tetramethylene ether) glycol, pentaerythritol, dipentaerythritol, tripentaerythritol, trimethylolpropane, neopentyl glycol, a sugar, preferably, sucrose, and mixtures thereof. 11. A lubricating member according to claim 1, wherein said metathesized unsaturated polyol ester is selected from the group consisting of metathesized Abyssinian oil, metathesized Almond Oil, metathesized Apricot Oil, metathesized Apricot Kernel oil, metathesized Argan oil, metathesized Avocado Oil, metathesized Babassu Oil, metathesized Baobab Oil, metathesized Black Cumin Oil, metathesized Black Currant Oil, metathesized Borage Oil, metathesized Camelina oil, metathesized Carinata oil, metathesized Canola oil, metathesized Castor oil, metathesized Cherry Kernel Oil, metathesized Coconut oil, metathesized Corn oil, metathesized Cottonseed oil, metathesized Echium Oil, metathesized Evening Primrose Oil, metathesized Flax Seed Oil, metathesized Grape Seed Oil, metathesized Grapefruit Seed Oil, metathesized Hazelnut Oil, metathesized Hemp Seed Oil, metathesized Jatropha oil, metathesized Jojoba Oil, metathesized Kukui Nut Oil, metathesized Linseed Oil, metathesized Macadamia Nut Oil, metathesized Meadowfoam Seed Oil, metathesized Moringa Oil, metathesized Neem Oil, metathesized Olive Oil, metathesized Palm Oil, metathesized Palm Kernel Oil, metathesized Peach Kernel Oil, metathesized Peanut Oil, metathesized Pecan Oil, metathesized Pennycress oil, metathesized Perilla Seed Oil, metathesized Pistachio Oil, metathesized Pomegranate Seed Oil, metathesized Pongamia oil, metathesized Pumpkin Seed Oil, metathesized Raspberry Oil, metathesized Red Palm Olein, metathesized Rice Bran Oil, metathesized Rosehip Oil, metathesized Safflower Oil, metathesized Seabuckthorn Fruit Oil, metathesized Sesame Seed Oil, metathesized Shea Olein, metathesized Sunflower Oil, metathesized Soybean Oil, metathesized Tonka Bean Oil, metathesized Tung Oil, metathesized Walnut Oil, metathesized Wheat Germ Oil, metathesized High Oleoyl Soybean Oil, metathesized High Oleoyl Sunflower Oil, metathesized High Oleoyl Safflower Oil, metathesized High Erucic Acid Rapeseed Oil, and mixtures thereof. 12. A lubricating member according to claim 1, further comprising a water soluble polymer. 13. The lubricating member according to claim 12, wherein said water soluble polymer is selected from polyethylene oxide, polyvinyl pyrrolidone, polyacrylamide, polyhydroxymethacrylate, polyvinyl imidazoline, polyethylene glycol, polyvinyl alcohol, polyhydroxyethymethacrylate, guars, cellulose, modified cellulose and mixtures thereof. 14. The lubricating member according to claim 12, wherein said water soluble polymer is polyethylene oxide having an average molecular weight of at least 300000, preferably from 300,000 to 8 million, more preferably from 1 million to 5 million, most preferably from 2 to 3 million. 15. The lubricating member according to claim 12, wherein said water soluble polymer further comprises from 0.01% to 50%, preferably from 2% to 40%, by weight of the lubricating material of a copolymer of polyethylene oxide and polypropylene oxide. 16. The lubricating member according to claim 12, wherein said polyethylene oxide polymer is present at a level of from 15% to 70%, preferably from 20% to 60%, more preferably from 25% to 50% by weight of the lubricating material. 17. The lubricating member according to claim 1, wherein said lubricating member further comprises from 1% to 50% by weight of a water insoluble material, preferably selected from polyethylene, polypropylene, polystyrene, high impact polystyrene, butadiene styrene copolymer, polyacetal, acrylonitrile-butadiene styrene copolymer, ethylene vinyl acetate copolymer and mixtures thereof. 18. A hair removal cartridge having a front end and an opposing rear end, the hair removal cartridge comprising:
a. at least one hair removal member positioned between said front end and said rear end; and b. at least one lubricating member according to claim 1. 19. A hair removal device comprising:
a. a hair removal cartridge according to claim 18, and b. a handle permanently or removably attached to said hair removal cartridge. | The invention relates to a lubricating member for a razor cartridge comprising a metathesized unsaturated polyol ester for improved lubrication.1. A lubricating member for use on a hair removal device, said lubricating member comprising a lubricating material comprising a) from 1% to 99% by weight of methathesized unsaturated polyol ester methathesized unsaturated polyol ester, having one or more of the following properties:
(i) a weight average molecular weight of from about 5,000 Daltons to about 50,000 Daltons; (ii) an oligomer index from greater than 0 to 1; and (iii) an iodine value of from about 30 to about 200. 2. A device according to claim 1, wherein said metathesized unsaturated polyol ester has a weight average molecular weight of from about 5,000 Daltons to about 50,000 Daltons. 3. A device according to claim 1 wherein said metathesized unsaturated polyol ester has an iodine value of from about 30 to about 200. 4. A lubricating member for use on a hair removal device, said lubricating member comprising a lubricating material comprising a) from 1% to 99% by weight of composition comprising:
a) a metathesized unsaturated polyol ester, said metathesized unsaturated polyol ester having a weight average molecular weight of from about 2,000 Daltons to about 50,000 Daltons; and one or more of the following properties:
(i) a free hydrocarbon content, based on total weight of metathesized unsaturated polyol ester of from about 0% to about 5%;
(ii) an oligomer index from greater than 0 to 1; and
(iii) an iodine value of from about 8 to about 200. 5. A lubricating member according to claim 4, wherein said metathesized unsaturated polyol ester has an iodine value of from about 10 to about 200. 6. A lubricating member according to claim 4, wherein said metathesized unsaturated polyol ester has an oligomer index from about 0.001 to 1. 7. A lubricating member according to claim 1, wherein said metathesized unsaturated polyol ester has a free hydrocarbon content, based on total weight of metathesized unsaturated polyol ester, of from about 0% to about 5%. 8. A lubricating member according to claim 1, said composition comprising, based on total composition weight, from about 0.1% to about 50% of said metathesized unsaturated polyol ester. 9. A lubricating member according to claim 1, wherein the metathesized unsaturated polyol ester is metathesized at least once. 10. A lubricating member according to claim 1, wherein said metathesized unsaturated polyol ester is derived from a natural polyol ester and/or a synthetic polyol ester, preferably said natural polyol ester is selected from the group consisting of a vegetable oil, an animal fat, an algae oil and mixtures thereof; and said synthetic polyol ester is derived from a material selected from the group consisting of ethylene glycol, propylene glycol, glycerol, polyglycerol, polyethylene glycol, polypropylene glycol, poly(tetramethylene ether) glycol, pentaerythritol, dipentaerythritol, tripentaerythritol, trimethylolpropane, neopentyl glycol, a sugar, preferably, sucrose, and mixtures thereof. 11. A lubricating member according to claim 1, wherein said metathesized unsaturated polyol ester is selected from the group consisting of metathesized Abyssinian oil, metathesized Almond Oil, metathesized Apricot Oil, metathesized Apricot Kernel oil, metathesized Argan oil, metathesized Avocado Oil, metathesized Babassu Oil, metathesized Baobab Oil, metathesized Black Cumin Oil, metathesized Black Currant Oil, metathesized Borage Oil, metathesized Camelina oil, metathesized Carinata oil, metathesized Canola oil, metathesized Castor oil, metathesized Cherry Kernel Oil, metathesized Coconut oil, metathesized Corn oil, metathesized Cottonseed oil, metathesized Echium Oil, metathesized Evening Primrose Oil, metathesized Flax Seed Oil, metathesized Grape Seed Oil, metathesized Grapefruit Seed Oil, metathesized Hazelnut Oil, metathesized Hemp Seed Oil, metathesized Jatropha oil, metathesized Jojoba Oil, metathesized Kukui Nut Oil, metathesized Linseed Oil, metathesized Macadamia Nut Oil, metathesized Meadowfoam Seed Oil, metathesized Moringa Oil, metathesized Neem Oil, metathesized Olive Oil, metathesized Palm Oil, metathesized Palm Kernel Oil, metathesized Peach Kernel Oil, metathesized Peanut Oil, metathesized Pecan Oil, metathesized Pennycress oil, metathesized Perilla Seed Oil, metathesized Pistachio Oil, metathesized Pomegranate Seed Oil, metathesized Pongamia oil, metathesized Pumpkin Seed Oil, metathesized Raspberry Oil, metathesized Red Palm Olein, metathesized Rice Bran Oil, metathesized Rosehip Oil, metathesized Safflower Oil, metathesized Seabuckthorn Fruit Oil, metathesized Sesame Seed Oil, metathesized Shea Olein, metathesized Sunflower Oil, metathesized Soybean Oil, metathesized Tonka Bean Oil, metathesized Tung Oil, metathesized Walnut Oil, metathesized Wheat Germ Oil, metathesized High Oleoyl Soybean Oil, metathesized High Oleoyl Sunflower Oil, metathesized High Oleoyl Safflower Oil, metathesized High Erucic Acid Rapeseed Oil, and mixtures thereof. 12. A lubricating member according to claim 1, further comprising a water soluble polymer. 13. The lubricating member according to claim 12, wherein said water soluble polymer is selected from polyethylene oxide, polyvinyl pyrrolidone, polyacrylamide, polyhydroxymethacrylate, polyvinyl imidazoline, polyethylene glycol, polyvinyl alcohol, polyhydroxyethymethacrylate, guars, cellulose, modified cellulose and mixtures thereof. 14. The lubricating member according to claim 12, wherein said water soluble polymer is polyethylene oxide having an average molecular weight of at least 300000, preferably from 300,000 to 8 million, more preferably from 1 million to 5 million, most preferably from 2 to 3 million. 15. The lubricating member according to claim 12, wherein said water soluble polymer further comprises from 0.01% to 50%, preferably from 2% to 40%, by weight of the lubricating material of a copolymer of polyethylene oxide and polypropylene oxide. 16. The lubricating member according to claim 12, wherein said polyethylene oxide polymer is present at a level of from 15% to 70%, preferably from 20% to 60%, more preferably from 25% to 50% by weight of the lubricating material. 17. The lubricating member according to claim 1, wherein said lubricating member further comprises from 1% to 50% by weight of a water insoluble material, preferably selected from polyethylene, polypropylene, polystyrene, high impact polystyrene, butadiene styrene copolymer, polyacetal, acrylonitrile-butadiene styrene copolymer, ethylene vinyl acetate copolymer and mixtures thereof. 18. A hair removal cartridge having a front end and an opposing rear end, the hair removal cartridge comprising:
a. at least one hair removal member positioned between said front end and said rear end; and b. at least one lubricating member according to claim 1. 19. A hair removal device comprising:
a. a hair removal cartridge according to claim 18, and b. a handle permanently or removably attached to said hair removal cartridge. | 1,600 |
1,001 | 15,198,234 | 1,612 | A consumer product including a fabric and home composition providing for multiple blooms of fragrance, the multiple blooms being provided for by different populations of microcapsules and method related thereto. | 1. A consumer product comprising a composition, the composition comprising:
an adjunct material; a first population of microcapsules, the first population having a first median volume weighted particle size and comprising microcapsules comprising a partitioning modifier and a first perfume oil at a first weight ratio; and a second population of microcapsules, the second population having a second median volume weighted particle size and comprising microcapsules comprising the partitioning modifier and a second perfume oil at a second weight ratio; wherein the first weight ratio and the second weight ratio are different, and/or the first median volume weighted particle size and the second median volume weighted particle size are different; wherein the composition is a fabric and home care composition. 2. The consumer product of claim 1, wherein the first weight ratio is a weight ratio of from 2:3 to 3:2 of the partitioning modifier to the first perfume oil; and wherein the second weight ratio is a weight ratio of greater than 0 to less than 2:3 of the partitioning modifier to the second perfume oil. 3. The consumer product of claim 1, wherein the weight ratio of the first population of microcapsules to the second population of microcapsules is greater than 0 to less than 1:1. 4. The consumer product of claim 1, wherein the weight ratio of the first population of microcapsules to the second population of microcapsules exceeds 1:1. 5. The consumer product of claim 1, wherein the median volume-weighted particle size is from 2 microns to 80 microns. 6. The consumer product of claim 1, wherein the first median volume weighted particle size is different from the second median volume weighted particle size. 7. The consumer product of claim 1, wherein the first perfume oil and the second perfume oil are the same. 8. The consumer product of claim 1, wherein the adjunct material comprises a non-encapsulated perfume oil. 9. The consumer product of claim 8, wherein the non-encapsulated perfume oil is different from the first and second perfume oil. 10. The consumer product of claim 1, wherein the partitioning modifier is selected from the group consisting of isopropyl myristate, mono-, di-, and tri-esters of C4-C24 fatty acids, castor oil, mineral oil, soybean oil, hexadecanoic acid, methyl ester isododecane, isoparaffin oil, polydimethylsiloxane, brominated vegetable oil, and mixtures thereof. 11. The consumer product of claim 1, wherein the microcapsules further comprise a shell material selected from the group consisting of polyacrylates, polyethylenes, polyamides, polystyrenes, polyisoprenes, polycarbonates, polyesters, polyureas, polyurethanes, polyolefins, polysaccharides, epoxy resins, vinyl polymers, urea cross-linked with formaldehyde or gluteraldehyde, melamine cross-linked with formaldehyde; gelatin-polyphosphate coacervates optionally cross-linked with gluteraldehyde; gelatin-gum Arabic coacervates; cross-linked silicone fluids; polyamine reacted with polyisocyanates; acrylate monomers polymerized via free radical polymerization, silk, wool, gelatine, cellulose, proteins, and mixtures thereof. 12. The consumer product of claim 1, wherein the microcapsules further comprise a shell material comprising a reaction product of a first substance in the presence of a second substance comprising an emulsifier, the first substance comprising a reaction product of i) an oil soluble or dispersible amine with ii) a multifunctional acrylate or methacrylate monomer or oligomer, an oil soluble acid and an initiator, the emulsifier comprising a water soluble or water dispersible acrylic acid alkyl acid copolymer, an alkali or alkali salt, and optionally a water phase initiator. 13. The consumer product of claim 1, wherein the adjunct material comprises at least one surfactant, the at least one surfactant comprising from about 0.01% to about 30% by weight of the composition. 14. The consumer product of claim 1, wherein the adjunct material comprises a fabric softening active. 15. The consumer product of claim 1, wherein the adjunct material comprises water. 16. The consumer product of claim 1, wherein the adjunct material comprises a builder. 17. The consumer product of claim 1, wherein the adjunct material comprises a material selected from the group consisting of dispersants, enzymes, dye transfer inhibiting agents, chelants, brighteners, stabilizers, silicones, fabric hueing agent, structurant, anti-agglomeration agent, and mixtures thereof. 18. The consumer product of claim 1, wherein adjunct material comprises a deposition aid. 19. The consumer product of claim 1, wherein the consumer product is in the form of a single unit dose or a multi-compartment unit dose. 20. The consumer product of claim 19, wherein the adjunct material comprises polyvinyl alcohol. 21. The consumer product of claim 1, wherein the consumer product is selected from the group consisting of all-purpose washing agents; heavy-duty washing agents; liquid fine-fabric detergents; hand dishwashing agents; light duty dishwashing agents; machine dishwashing agents, liquid cleaning and disinfecting agents; fabric conditioning products; cleaning auxiliaries; substrate-laden products; sprays; and mists. 22. The consumer product of claim 1, wherein the first perfume oil and the second perfume oil comprise at least one different material. 23. The consumer product of claim 1, wherein the first population and the second population comprise different shell materials. | A consumer product including a fabric and home composition providing for multiple blooms of fragrance, the multiple blooms being provided for by different populations of microcapsules and method related thereto.1. A consumer product comprising a composition, the composition comprising:
an adjunct material; a first population of microcapsules, the first population having a first median volume weighted particle size and comprising microcapsules comprising a partitioning modifier and a first perfume oil at a first weight ratio; and a second population of microcapsules, the second population having a second median volume weighted particle size and comprising microcapsules comprising the partitioning modifier and a second perfume oil at a second weight ratio; wherein the first weight ratio and the second weight ratio are different, and/or the first median volume weighted particle size and the second median volume weighted particle size are different; wherein the composition is a fabric and home care composition. 2. The consumer product of claim 1, wherein the first weight ratio is a weight ratio of from 2:3 to 3:2 of the partitioning modifier to the first perfume oil; and wherein the second weight ratio is a weight ratio of greater than 0 to less than 2:3 of the partitioning modifier to the second perfume oil. 3. The consumer product of claim 1, wherein the weight ratio of the first population of microcapsules to the second population of microcapsules is greater than 0 to less than 1:1. 4. The consumer product of claim 1, wherein the weight ratio of the first population of microcapsules to the second population of microcapsules exceeds 1:1. 5. The consumer product of claim 1, wherein the median volume-weighted particle size is from 2 microns to 80 microns. 6. The consumer product of claim 1, wherein the first median volume weighted particle size is different from the second median volume weighted particle size. 7. The consumer product of claim 1, wherein the first perfume oil and the second perfume oil are the same. 8. The consumer product of claim 1, wherein the adjunct material comprises a non-encapsulated perfume oil. 9. The consumer product of claim 8, wherein the non-encapsulated perfume oil is different from the first and second perfume oil. 10. The consumer product of claim 1, wherein the partitioning modifier is selected from the group consisting of isopropyl myristate, mono-, di-, and tri-esters of C4-C24 fatty acids, castor oil, mineral oil, soybean oil, hexadecanoic acid, methyl ester isododecane, isoparaffin oil, polydimethylsiloxane, brominated vegetable oil, and mixtures thereof. 11. The consumer product of claim 1, wherein the microcapsules further comprise a shell material selected from the group consisting of polyacrylates, polyethylenes, polyamides, polystyrenes, polyisoprenes, polycarbonates, polyesters, polyureas, polyurethanes, polyolefins, polysaccharides, epoxy resins, vinyl polymers, urea cross-linked with formaldehyde or gluteraldehyde, melamine cross-linked with formaldehyde; gelatin-polyphosphate coacervates optionally cross-linked with gluteraldehyde; gelatin-gum Arabic coacervates; cross-linked silicone fluids; polyamine reacted with polyisocyanates; acrylate monomers polymerized via free radical polymerization, silk, wool, gelatine, cellulose, proteins, and mixtures thereof. 12. The consumer product of claim 1, wherein the microcapsules further comprise a shell material comprising a reaction product of a first substance in the presence of a second substance comprising an emulsifier, the first substance comprising a reaction product of i) an oil soluble or dispersible amine with ii) a multifunctional acrylate or methacrylate monomer or oligomer, an oil soluble acid and an initiator, the emulsifier comprising a water soluble or water dispersible acrylic acid alkyl acid copolymer, an alkali or alkali salt, and optionally a water phase initiator. 13. The consumer product of claim 1, wherein the adjunct material comprises at least one surfactant, the at least one surfactant comprising from about 0.01% to about 30% by weight of the composition. 14. The consumer product of claim 1, wherein the adjunct material comprises a fabric softening active. 15. The consumer product of claim 1, wherein the adjunct material comprises water. 16. The consumer product of claim 1, wherein the adjunct material comprises a builder. 17. The consumer product of claim 1, wherein the adjunct material comprises a material selected from the group consisting of dispersants, enzymes, dye transfer inhibiting agents, chelants, brighteners, stabilizers, silicones, fabric hueing agent, structurant, anti-agglomeration agent, and mixtures thereof. 18. The consumer product of claim 1, wherein adjunct material comprises a deposition aid. 19. The consumer product of claim 1, wherein the consumer product is in the form of a single unit dose or a multi-compartment unit dose. 20. The consumer product of claim 19, wherein the adjunct material comprises polyvinyl alcohol. 21. The consumer product of claim 1, wherein the consumer product is selected from the group consisting of all-purpose washing agents; heavy-duty washing agents; liquid fine-fabric detergents; hand dishwashing agents; light duty dishwashing agents; machine dishwashing agents, liquid cleaning and disinfecting agents; fabric conditioning products; cleaning auxiliaries; substrate-laden products; sprays; and mists. 22. The consumer product of claim 1, wherein the first perfume oil and the second perfume oil comprise at least one different material. 23. The consumer product of claim 1, wherein the first population and the second population comprise different shell materials. | 1,600 |
1,002 | 15,100,990 | 1,612 | A composition comprising a zinc ion source for use in inhibiting the adherence to or invasion of oral soft tissue, such as gingival epithelial cells, by a periodontal pathogen is provided. | 1. A composition comprising a zinc ion source for use in inhibiting the adherence to or invasion of oral soft tissue epithelial cells by a periodontal pathogen. 2. The composition of claim 1, wherein the oral soft tissue is gingival epithelial cells. 3. The composition of claim 1, wherein the periodontal pathogen causes, transmits or exacerbates a periodontal disease selected from gingivitis and periodontitis. 4. The composition of claim 1, wherein the periodontal pathogen is one or more of Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguis, Streptococcus oralis, Porphyromonos gingivalis, Porphyromonas congingivalis, Actinobacillus actinomycetemcomitans, Tannerella forsythia, Fusobacterium nucleatum and Treponema denticola. 5. The composition of claim 1 wherein the periodontal pathogen is Porphyromonas gingivalis and/or Actinobacillus actinomycetemcomitans. 6. The composition according to claim 1 wherein the zinc ion source comprises one or more zinc salts selected from zinc chloride, zinc, acetate, zinc gluconate, zinc, sulphate, zinc fluoride, zinc citrate, zinc lactate, zinc, oxide, zinc monoglycerolate, zinc tartrate, zinc pyrophosphate, zinc phosphate, zinc maleate, zinc malate, zinc carbonate, zinc ascorbate, zinc lysine hydrochloride and zinc chloride hydroxide monohydrate (TBZC). 7. The composition of claim 1 wherein the zinc ion source comprises one or more zinc salts selected from zinc citrate and zinc oxide. 8. The composition claim 1 wherein the composition comprises between 1 and 20,000 ppm zinc. 9. The composition of claim 1 wherein the composition delivers between 1 and 50 μM zinc to a surface in the oral cavity. 10. The composition of claim 1 wherein the composition delivers between 10 and 50 μM zinc to a surface in the oral cavity. 11. The composition of claim 1 for use in a mammalian subject. 12. The composition of claim 1 for use in a human subject. 13. The composition of claim 1 wherein the composition is an oral care composition. 14. The composition of claim 1 wherein the composition is an oral care composition selected from dentifrices, toothpastes, tooth powders, mouth rinses, lozenges, gums, gels, paints, and films. 15. A method for inhibiting the adherence to or invasion of oral soft tissue epithelial cells by a periodontal pathogen, the method comprising applying a composition comprising a zinc ion source to the oral soft tissue epithelial cells. 16. The method of claim 15, wherein the oral soft tissue is gingival epithelial cells. 17. The method of claim 15 wherein the periodontal pathogen causes transmits or exacerbates a periodontal disease selected from gingivitis and periodontitis. 18. The method of claim 15 wherein the periodontal pathogen is one or more of Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguis, Streptococcus oralis, Porphyromonos gingivalis, Porphyromonas congingivalis, Actinobacillus actinomycetemcomitans, Tannerella forsythia, Fusobacterium nucleatum and Treponema denticola. 19. The method of claim 15 wherein the periodontal pathogen is Porphyromonas gingivalis and/or Actinobacillus actinomycetemcomitans. 20. The method of claim 15 wherein the zinc ion source comprises one or more zinc salts selected from zinc chloride, zinc acetate, zinc gluconate, zinc, sulphate, zinc fluoride, zinc citrate, zinc lactate, zinc oxide, zinc monoglycerolate, zinc tartrate, zinc pyrophosphate. zinc phosphate, zinc maleate, zinc malate, zinc carbonate, zinc ascorbate, zinc lysine hydrochloride and zinc chloride hydroxide monohydrate (TBZC). 21. The method of claim 15 wherein the zinc ion source comprises one or more zinc salts selected from zinc citrate and zinc oxide. 22. The method of claim 15 wherein the composition comprises between 1 and 20,000 ppm zinc. 23. The method of claim 15 wherein the composition delivers between 1 and 50 μM zinc to a surface in the and cavity. 24. The method of claim 15 wherein the composition delivers between 10 and 50 μM zinc to a surface in the oral cavity. 25. The method of claim 15 wherein the composition is applied to gingival epithelial tissue of a mammal. 26. The method of claim 15 wherein the composition is applied to gingival epithelial tissue of a human. 27. The method of claim 15 wherein the composition is applied at least once per day. 28-42. (canceled) | A composition comprising a zinc ion source for use in inhibiting the adherence to or invasion of oral soft tissue, such as gingival epithelial cells, by a periodontal pathogen is provided.1. A composition comprising a zinc ion source for use in inhibiting the adherence to or invasion of oral soft tissue epithelial cells by a periodontal pathogen. 2. The composition of claim 1, wherein the oral soft tissue is gingival epithelial cells. 3. The composition of claim 1, wherein the periodontal pathogen causes, transmits or exacerbates a periodontal disease selected from gingivitis and periodontitis. 4. The composition of claim 1, wherein the periodontal pathogen is one or more of Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguis, Streptococcus oralis, Porphyromonos gingivalis, Porphyromonas congingivalis, Actinobacillus actinomycetemcomitans, Tannerella forsythia, Fusobacterium nucleatum and Treponema denticola. 5. The composition of claim 1 wherein the periodontal pathogen is Porphyromonas gingivalis and/or Actinobacillus actinomycetemcomitans. 6. The composition according to claim 1 wherein the zinc ion source comprises one or more zinc salts selected from zinc chloride, zinc, acetate, zinc gluconate, zinc, sulphate, zinc fluoride, zinc citrate, zinc lactate, zinc, oxide, zinc monoglycerolate, zinc tartrate, zinc pyrophosphate, zinc phosphate, zinc maleate, zinc malate, zinc carbonate, zinc ascorbate, zinc lysine hydrochloride and zinc chloride hydroxide monohydrate (TBZC). 7. The composition of claim 1 wherein the zinc ion source comprises one or more zinc salts selected from zinc citrate and zinc oxide. 8. The composition claim 1 wherein the composition comprises between 1 and 20,000 ppm zinc. 9. The composition of claim 1 wherein the composition delivers between 1 and 50 μM zinc to a surface in the oral cavity. 10. The composition of claim 1 wherein the composition delivers between 10 and 50 μM zinc to a surface in the oral cavity. 11. The composition of claim 1 for use in a mammalian subject. 12. The composition of claim 1 for use in a human subject. 13. The composition of claim 1 wherein the composition is an oral care composition. 14. The composition of claim 1 wherein the composition is an oral care composition selected from dentifrices, toothpastes, tooth powders, mouth rinses, lozenges, gums, gels, paints, and films. 15. A method for inhibiting the adherence to or invasion of oral soft tissue epithelial cells by a periodontal pathogen, the method comprising applying a composition comprising a zinc ion source to the oral soft tissue epithelial cells. 16. The method of claim 15, wherein the oral soft tissue is gingival epithelial cells. 17. The method of claim 15 wherein the periodontal pathogen causes transmits or exacerbates a periodontal disease selected from gingivitis and periodontitis. 18. The method of claim 15 wherein the periodontal pathogen is one or more of Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguis, Streptococcus oralis, Porphyromonos gingivalis, Porphyromonas congingivalis, Actinobacillus actinomycetemcomitans, Tannerella forsythia, Fusobacterium nucleatum and Treponema denticola. 19. The method of claim 15 wherein the periodontal pathogen is Porphyromonas gingivalis and/or Actinobacillus actinomycetemcomitans. 20. The method of claim 15 wherein the zinc ion source comprises one or more zinc salts selected from zinc chloride, zinc acetate, zinc gluconate, zinc, sulphate, zinc fluoride, zinc citrate, zinc lactate, zinc oxide, zinc monoglycerolate, zinc tartrate, zinc pyrophosphate. zinc phosphate, zinc maleate, zinc malate, zinc carbonate, zinc ascorbate, zinc lysine hydrochloride and zinc chloride hydroxide monohydrate (TBZC). 21. The method of claim 15 wherein the zinc ion source comprises one or more zinc salts selected from zinc citrate and zinc oxide. 22. The method of claim 15 wherein the composition comprises between 1 and 20,000 ppm zinc. 23. The method of claim 15 wherein the composition delivers between 1 and 50 μM zinc to a surface in the and cavity. 24. The method of claim 15 wherein the composition delivers between 10 and 50 μM zinc to a surface in the oral cavity. 25. The method of claim 15 wherein the composition is applied to gingival epithelial tissue of a mammal. 26. The method of claim 15 wherein the composition is applied to gingival epithelial tissue of a human. 27. The method of claim 15 wherein the composition is applied at least once per day. 28-42. (canceled) | 1,600 |
1,003 | 13,814,009 | 1,653 | Methods, devices, and systems are provided for guiding tumor movement, particularly in vivo for treatment of patients. The method may include implanting into a tissue site where tumor cells are present a device having one or more surface structures or substrates, such as aligned nanofibers, which provide physical guidance cues for directing the migration of the tumor cells from the first tissue location to a selected second location, for tumor cell extraction or death. The devices and systems may include a cytotoxic agent for contacting tumor cells migrated via the substrate. All or a portion of the at least one substrate may include one or more biochemical cues, such as a coating of laminin or another protein, which may be provided in a concentration gradient to facilitate uni-directional tumor cell migration. | 1. An implantable system for promoting tumor cell migration for cell removal or death, the system comprising:
at least one substrate having a surface configured to provide cues for directing tumor cell migration along the substrate surface, the at least one substrate comprising a plurality of aligned nanofibers; and at least one cytotoxic agent for contacting tumor cells migrated via the at least one substrate. 2. The system of claim 1, wherein the plurality of aligned nanofibers form a tubular construct. 3. The system of claim 1, wherein the plurality of aligned nanofibers are disposed in a tubular construct. 4. The system of claim 3, wherein the tubular construct comprises an annular tube formed from a polycaprolactone, a polyurethane, or a combination thereof. 5. The system of claim 1, wherein the at least one cytotoxic agent is tethered or conjugated to at least a portion of the plurality of aligned nanofibers. 6. The system of claim 1, wherein the at least one cytotoxic agent is tethered or conjugated to a polymeric sink material. 7. The system of claim 6, wherein the polymeric sink material comprises a hydrogel. 8. The system of claim 1, wherein the cytotoxic agent comprises cyclopamine, honokiol, furegrelate, doxorubicin, or a combination thereof. 9. The system of claim 1, wherein all or a portion of the at least one substrate comprises one or more biochemical cues to facilitate uni-directional tumor cell migration. 10. The system of claim 1, wherein the plurality of nanofibers comprises a coating selected from the group consisting of extracellular matrix proteins, growth factors, cytokines, peptides, and combinations thereof. 11. The system of claim 10, wherein the coating is disposed uniformly along the length of the plurality of aligned nanofibers. 12. The system of claim 10, wherein the coating is disposed in a concentration gradient along the length of the plurality of aligned nanofibers from a first end of the plurality of aligned nanofibers to a second end of the plurality of aligned nanofibers, the second end being distal to the first end. 13. The system of claim 12, wherein the concentration gradient is effective to promote uni-directional migration of tumor cells. 14. The system of claim 12, wherein the concentration gradient is effective to promote bi-directional migration of non-tumor cells. 15. The system of claim 1, wherein the nanofibers comprise a synthetic polymer. 16. The system of any one of claims 9 to 15, wherein the nanofiber films comprise a coating of myelin or a basement membrane protein. 17. The system of claim 16, wherein the nanofiber films comprise a laminin coating. 18. The system of claim 1, wherein the nanofibers have a diameter in the range of about 400 nm to about 800 nm. 19. The system of claim 1, wherein the plurality of nanofibers are in the form of a nanofiber film having a thickness in the range of 5 microns to 20 microns. 20. The system of any one of claims 1 to 19, wherein the plurality of aligned nanofibers are synthetic polymeric fibers having a diameter from 600 nm to 800 nm, which are coated with an extracellular matrix protein having a concentration gradient along the length of the fibers. 21. The system of claim 20, wherein the cytotoxic agent comprises collagen hydrogel covalently coupled to an apoptotic triggering agent. 22. The system of claim 21, wherein the hydrogel is at least partially encased in a cell-impermeable pouch. 23. An implantable device for promoting tumor cell migration for cell removal or death, the device comprising:
at least one film having a surface configured to provide cues for directing tumor cell migration along the substrate surface, wherein the surface comprises a coating material gradient to effect uni-directional or bi-direction growth of cells across the surface. 24. The device of claim 23, further comprising a cytotoxic agent positioned for contacting tumor cells migrated via the surface. 30. A method for guiding tumor movement in vivo, comprising:
implanting into a tissue site where tumor cells are present a device having one or more surface structures which provide physical guidance cues for directing the migration of the tumor cells from the first tissue location to a selected second location. 31. The method of claim 30, wherein the surface structure comprises nanofibers or grooves. 32. The method of claim 30 or 31, further comprising applying another guidance means at the tissue implantation site, wherein the guidance means comprises an electric field, one or more biochemical cue, or cell seeding, and works in combination with the physical guidance cues of the surface structure to direct migration of the tumor cells to the selected second location. 33. The method of claim 30 or 31, wherein the surface structure comprises a plurality of aligned nanofibers having a diameter from 400 nm to 800 nm, which are coated with an extracellular matrix protein. 34. The method of any one of claims 30 to 33, wherein the tumor cells comprise Medulloblastoma cells or malignant glioma cells. 35. A method for treating a patient comprising:
implanting at a tissue site in the patient a device comprising a substrate having a surface configured to provide cues for directing tumor cell migration along the substrate surface, the substrate comprising a plurality of aligned nanofibers; and subsequently killing or removing tumor cells that have migrated along the substrate surface. 36. The method of claim 35, wherein the tissue site comprises a tumor or a void of a resected tumor. 37. The method of claim 35, further comprising contacting tumor cells that have migrated along at least a portion of the substrate with a cytotoxic agent. 38. The method of claim 37, wherein the cytotoxic agent is part of the implanted device. 39. The method of claim any one of claims 35 to 38, wherein tumor cells are relocated from a first region to a second region from a primary tumor via the implantable device before the tumor cells are killed or removed. | Methods, devices, and systems are provided for guiding tumor movement, particularly in vivo for treatment of patients. The method may include implanting into a tissue site where tumor cells are present a device having one or more surface structures or substrates, such as aligned nanofibers, which provide physical guidance cues for directing the migration of the tumor cells from the first tissue location to a selected second location, for tumor cell extraction or death. The devices and systems may include a cytotoxic agent for contacting tumor cells migrated via the substrate. All or a portion of the at least one substrate may include one or more biochemical cues, such as a coating of laminin or another protein, which may be provided in a concentration gradient to facilitate uni-directional tumor cell migration.1. An implantable system for promoting tumor cell migration for cell removal or death, the system comprising:
at least one substrate having a surface configured to provide cues for directing tumor cell migration along the substrate surface, the at least one substrate comprising a plurality of aligned nanofibers; and at least one cytotoxic agent for contacting tumor cells migrated via the at least one substrate. 2. The system of claim 1, wherein the plurality of aligned nanofibers form a tubular construct. 3. The system of claim 1, wherein the plurality of aligned nanofibers are disposed in a tubular construct. 4. The system of claim 3, wherein the tubular construct comprises an annular tube formed from a polycaprolactone, a polyurethane, or a combination thereof. 5. The system of claim 1, wherein the at least one cytotoxic agent is tethered or conjugated to at least a portion of the plurality of aligned nanofibers. 6. The system of claim 1, wherein the at least one cytotoxic agent is tethered or conjugated to a polymeric sink material. 7. The system of claim 6, wherein the polymeric sink material comprises a hydrogel. 8. The system of claim 1, wherein the cytotoxic agent comprises cyclopamine, honokiol, furegrelate, doxorubicin, or a combination thereof. 9. The system of claim 1, wherein all or a portion of the at least one substrate comprises one or more biochemical cues to facilitate uni-directional tumor cell migration. 10. The system of claim 1, wherein the plurality of nanofibers comprises a coating selected from the group consisting of extracellular matrix proteins, growth factors, cytokines, peptides, and combinations thereof. 11. The system of claim 10, wherein the coating is disposed uniformly along the length of the plurality of aligned nanofibers. 12. The system of claim 10, wherein the coating is disposed in a concentration gradient along the length of the plurality of aligned nanofibers from a first end of the plurality of aligned nanofibers to a second end of the plurality of aligned nanofibers, the second end being distal to the first end. 13. The system of claim 12, wherein the concentration gradient is effective to promote uni-directional migration of tumor cells. 14. The system of claim 12, wherein the concentration gradient is effective to promote bi-directional migration of non-tumor cells. 15. The system of claim 1, wherein the nanofibers comprise a synthetic polymer. 16. The system of any one of claims 9 to 15, wherein the nanofiber films comprise a coating of myelin or a basement membrane protein. 17. The system of claim 16, wherein the nanofiber films comprise a laminin coating. 18. The system of claim 1, wherein the nanofibers have a diameter in the range of about 400 nm to about 800 nm. 19. The system of claim 1, wherein the plurality of nanofibers are in the form of a nanofiber film having a thickness in the range of 5 microns to 20 microns. 20. The system of any one of claims 1 to 19, wherein the plurality of aligned nanofibers are synthetic polymeric fibers having a diameter from 600 nm to 800 nm, which are coated with an extracellular matrix protein having a concentration gradient along the length of the fibers. 21. The system of claim 20, wherein the cytotoxic agent comprises collagen hydrogel covalently coupled to an apoptotic triggering agent. 22. The system of claim 21, wherein the hydrogel is at least partially encased in a cell-impermeable pouch. 23. An implantable device for promoting tumor cell migration for cell removal or death, the device comprising:
at least one film having a surface configured to provide cues for directing tumor cell migration along the substrate surface, wherein the surface comprises a coating material gradient to effect uni-directional or bi-direction growth of cells across the surface. 24. The device of claim 23, further comprising a cytotoxic agent positioned for contacting tumor cells migrated via the surface. 30. A method for guiding tumor movement in vivo, comprising:
implanting into a tissue site where tumor cells are present a device having one or more surface structures which provide physical guidance cues for directing the migration of the tumor cells from the first tissue location to a selected second location. 31. The method of claim 30, wherein the surface structure comprises nanofibers or grooves. 32. The method of claim 30 or 31, further comprising applying another guidance means at the tissue implantation site, wherein the guidance means comprises an electric field, one or more biochemical cue, or cell seeding, and works in combination with the physical guidance cues of the surface structure to direct migration of the tumor cells to the selected second location. 33. The method of claim 30 or 31, wherein the surface structure comprises a plurality of aligned nanofibers having a diameter from 400 nm to 800 nm, which are coated with an extracellular matrix protein. 34. The method of any one of claims 30 to 33, wherein the tumor cells comprise Medulloblastoma cells or malignant glioma cells. 35. A method for treating a patient comprising:
implanting at a tissue site in the patient a device comprising a substrate having a surface configured to provide cues for directing tumor cell migration along the substrate surface, the substrate comprising a plurality of aligned nanofibers; and subsequently killing or removing tumor cells that have migrated along the substrate surface. 36. The method of claim 35, wherein the tissue site comprises a tumor or a void of a resected tumor. 37. The method of claim 35, further comprising contacting tumor cells that have migrated along at least a portion of the substrate with a cytotoxic agent. 38. The method of claim 37, wherein the cytotoxic agent is part of the implanted device. 39. The method of claim any one of claims 35 to 38, wherein tumor cells are relocated from a first region to a second region from a primary tumor via the implantable device before the tumor cells are killed or removed. | 1,600 |
1,004 | 14,786,980 | 1,623 | Phosphorylated hydrogels obtained by co-cross-linking hyaluronic acid with dextran, a process for preparing same, and a use of the hydrogel for the encapsulation and extended release of active principles as well as cells for use in regenerative human and veterinary medicine. | 1-12. (canceled) 13. A co-cross-linked phosphorylated polysaccharide hydrogel, comprising:
dextran; and hyaluronic acid and/or one of its salts, optionally functionalized, wherein said dextran and said hyaluronic acid and/or its salt, optionally functionalized, are bound together by phosphodiester and/or polyphosphodiester covalent bonds. 14. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 13, wherein the hyaluronic acid or its salt is a hyaluronic acid and/or its salt functionalized by sodium trimetaphosphate. 15. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 14, wherein the phosphate concentration of the hyaluronic acid and/or its salt functionalized by sodium trimetaphosphate is between 0.1 and 2 mEq/g. 16. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 13, wherein the phosphate concentration of said hydrogel is between 0.1 and 2 mEq/g. 17. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 13, wherein the co-cross-linked phosphorylated polysaccharide hydrogel is used to prepare a drug to be implanted alone. 18. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is associated with cells for cell therapy. 19. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is in wet form for topical application. 20. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is to deliver active molecules and active principles. 21. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 20, wherein the active molecules and active principles are chosen from:
fibroblast growth factors (FGFs); and/or platelet derived growth factors (PDGFs); and/or transformation growth factors (TGFs); and/or vascular endothelium growth factors (vEGF); and/or osteoinductive growth factors (BMPs). 22. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is to be used in regenerative medicine. 23. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is for use in human and/or veterinary regenerative medicine. 24. A process for preparing a co-cross-linked phosphorylated polysaccharide hydrogel, based on dextran and hyaluronic acid and/or one of its salts, optionally functionalized, the process comprising:
a) providing a dextran solution; b) activating at least one hydroxyl group of the dextran by adding an alkaline hydroxide solution to said dextran solution to obtain an activated dextran solution, wherein the concentration of alkaline hydroxide added is between 0.1 M and 0.5 M; c) adding sodium trimetaphosphate to said activated dextran solution; and d) adding hyaluronic acid and/or one of its salts, optionally functionalized, to a solution obtained in step c), wherein said steps a) through d) are performed at a temperature of between 18 and 25° C. 25. The process of claim 24, wherein steps c) and d) are performed concomitantly. 26. The process of claim 25, wherein the sodium trimetaphosphate and hyaluronic acid and/or one of its salts, optionally functionalized, is added concomitantly to the solution obtained in step b). 27. The process of claim 24, wherein the hyaluronic acid and/or one of its salts, optionally functionalized, added in step d) is a hyaluronic acid functionalized by sodium trimetaphosphate. 28. The process of claim 24, wherein said hyaluronic acid and/or one of its salts, optionally functionalized, is added in the form of a powder. 29. The process of claim 24, wherein the alkaline hydroxide solution comprises sodium hydroxide. 30. Use of the hydrogel claim 13 for osteoconductivity and/or osteoinductivity. | Phosphorylated hydrogels obtained by co-cross-linking hyaluronic acid with dextran, a process for preparing same, and a use of the hydrogel for the encapsulation and extended release of active principles as well as cells for use in regenerative human and veterinary medicine.1-12. (canceled) 13. A co-cross-linked phosphorylated polysaccharide hydrogel, comprising:
dextran; and hyaluronic acid and/or one of its salts, optionally functionalized, wherein said dextran and said hyaluronic acid and/or its salt, optionally functionalized, are bound together by phosphodiester and/or polyphosphodiester covalent bonds. 14. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 13, wherein the hyaluronic acid or its salt is a hyaluronic acid and/or its salt functionalized by sodium trimetaphosphate. 15. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 14, wherein the phosphate concentration of the hyaluronic acid and/or its salt functionalized by sodium trimetaphosphate is between 0.1 and 2 mEq/g. 16. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 13, wherein the phosphate concentration of said hydrogel is between 0.1 and 2 mEq/g. 17. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 13, wherein the co-cross-linked phosphorylated polysaccharide hydrogel is used to prepare a drug to be implanted alone. 18. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is associated with cells for cell therapy. 19. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is in wet form for topical application. 20. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is to deliver active molecules and active principles. 21. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 20, wherein the active molecules and active principles are chosen from:
fibroblast growth factors (FGFs); and/or platelet derived growth factors (PDGFs); and/or transformation growth factors (TGFs); and/or vascular endothelium growth factors (vEGF); and/or osteoinductive growth factors (BMPs). 22. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is to be used in regenerative medicine. 23. The co-cross-linked phosphorylated polysaccharide hydrogel of claim 17, wherein the drug is for use in human and/or veterinary regenerative medicine. 24. A process for preparing a co-cross-linked phosphorylated polysaccharide hydrogel, based on dextran and hyaluronic acid and/or one of its salts, optionally functionalized, the process comprising:
a) providing a dextran solution; b) activating at least one hydroxyl group of the dextran by adding an alkaline hydroxide solution to said dextran solution to obtain an activated dextran solution, wherein the concentration of alkaline hydroxide added is between 0.1 M and 0.5 M; c) adding sodium trimetaphosphate to said activated dextran solution; and d) adding hyaluronic acid and/or one of its salts, optionally functionalized, to a solution obtained in step c), wherein said steps a) through d) are performed at a temperature of between 18 and 25° C. 25. The process of claim 24, wherein steps c) and d) are performed concomitantly. 26. The process of claim 25, wherein the sodium trimetaphosphate and hyaluronic acid and/or one of its salts, optionally functionalized, is added concomitantly to the solution obtained in step b). 27. The process of claim 24, wherein the hyaluronic acid and/or one of its salts, optionally functionalized, added in step d) is a hyaluronic acid functionalized by sodium trimetaphosphate. 28. The process of claim 24, wherein said hyaluronic acid and/or one of its salts, optionally functionalized, is added in the form of a powder. 29. The process of claim 24, wherein the alkaline hydroxide solution comprises sodium hydroxide. 30. Use of the hydrogel claim 13 for osteoconductivity and/or osteoinductivity. | 1,600 |
1,005 | 14,770,862 | 1,628 | Compositions comprising 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof; a triglyceride; and a surfactant, as well as methods for their use. The present disclosure further relates to self-emulsifying drug delivery systems, such as SEDDS, SMEDDS, or SNEDDS comprising 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid. | 1-138. (canceled) 139. A composition comprising
from 5% to 60% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof; from 15% to 60% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 10% to 60% by weight of the total composition of a nonionic surfactant. 140. The composition according to claim 139, wherein the composition is a preconcentrate forming a nanoemulsion or microemulsion in contact with aqueous media. 141. The composition according to claim 139, further comprising a co-solvent. 142. The composition according to claim 139, wherein the 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof is present as its (R)-enantiomer, (S)-enantiomer, or a mixture of its (R)- and (S)-enantiomers. 143. The composition according to claim 139, comprising 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid
its (R)-enantiomer
its (S)-enantiomer
or
a mixture of its (R)- and (S)-enantiomer. 144. The composition according to claim 139, wherein the salt is formed from 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoate and an ion chosen from Li+, Na+, K+, NH4 +, Mg2+, Ca2+, and quaternized ions of meglumine, tris(hydroxymethyl)aminomethane, diethylamine, arginine, ethylenediamine, piperazine, and protonated chitosan salts. 145. The composition according to claim 139, wherein 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid is present as an ester derivative chosen from a C1-C6-alkyl or phenyl ester. 146. The composition according to claim 139, wherein the medium-chain triglyceride oil comprises C8-C10 triglycerides. 147. The composition according to claim 146, wherein the medium-chain triglyceride is derived from capric acid and caprylic acid. 148. The composition according to claim 146, wherein the medium-chain triglyceride oil comprises between 50% and 80% caprylic acid and between 20% to 50% capric acid or the medium-chain triglyceride oil comprises between about 50% to 60% caprylic acid and about 40% to 50% capric acid. 149. The composition according to claim 139, wherein the surfactant is chosen from polysorbates, sorbitan esters, ((poly(ethylene oxide))-(poly(propylene oxide)) (PEO-PPO)-block copolymers, polyoxyethylene (POE) castor oil, polyoxyethylene (POE) hydrogenated castor oil, polyoxyethylene (POE)-60-hydrogenated castor oil, polyoxyethylene (POE)-stearate, phospholipids, and mixtures thereof; from polyethylene (POE)-20-sorbitan monooleate, polyoxyethylene (20) sorbitan monooleate with a hydrophile-lipophile balance (HLB) value of 15.0 (TWEEN 80, CRILLET 4), polyoxyethylene (POE)-20-sorbitan monolaurate, polyoxyethylene sorbitan monostearate with an HLB value of 14.9 (TWEEN 60), polyoxyethylene (20) sorbitan monopalmitate with an HLB value of 15.6 (TWEEN 40), pharmaceutical grade polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20, CRILLET 1), Poloxamer 188 (PLURONIC F68, LUTROL F68), polyoxyethylene (POE) alkyl ethers, polyoxyethylene (POE)-10-oleyl ether, and polyethylene glycol oleyl ether with an HLB value of 12.4 (BRIJ 96 V), and mixtures thereof; from 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15), polyoxyethylated castor oil with a hydrophile-lipophile balance (HLB) value of 2-14 (CREMOPHOR EL), hydrogenated polyoxyethylated castor oil with an HLB value of 14-16 (CREMOPHOR RH40), polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), polyoxyethylene (20) sorbitan monopalmitate with an HLB value of 15.6 (TWEEN 40), polyoxyethylene sorbitan monostearate with an HLB value of 14.9 (TWEEN 60), polyoxyethylene (20) sorbitan monooleate with an HLB value of 15.0 (TWEEN 80), sorbitan monooleate with an HLB value of 4.3 (SPAN 80), polyoxyethylene (10) oleyl ether with an HLB value of 12 (BRIJ 97), and mixtures thereof. 150. The composition according to claim 141, comprising from 0% to 20% or from 0% to 10% by weight of the total composition of co-solvent and wherein the co-solvent is chosen from C1 to C6 alcohols, ethanol, benzyl alcohol, alkane diols and triols, glycol ethers, propylene glycol (PG), glycerol, tetraglycol, polyethylene glycols (PEG), PEG 400, pyrrolidine derivatives, 2-pyrrolidone, triacetin, and mixtures thereof. 151. The composition according to claim 139, comprising:
2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, an MCT oil, and a surfactant chosen from polysorbates, and a co-solvent PEG 400; 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, an MCT oil, polyoxyethylene (20) sorbitan monooleate with an HLB value of 15.0 (TWEEN 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15), and PEG 400; 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, an MCT oil, polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), polyoxyethylene (20) sorbitan monooleate with an HLB value of 15.0 (TWEEN 80), and 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15); 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, a MCT oil, polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15), and PEG 400; 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, a MCT oil, and polyoxyethylene (20) sorbitan monooleate with an HLB value of 15.0 (TWEEN 80); 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, a MCT oil, and polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20); or 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), and a medium-chain triglyceride oil comprising between about 50% to 60% caprylic acid and about 40% to 50% capric acid (MIGLYOL 812 N). 152. The composition according to claim 139, comprising:
from 5% to 46% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid; from 20% to 49% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 30% to 60% by weight of the total composition of a nonionic surfactant; from 15% to 48% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid; from 20% to 40% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 30% to 50% by weight of the total composition of a nonionic surfactant; from 40% to 48% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid; from 20% to 30% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 30% to 40% by weight of the total composition of a nonionic surfactant; or from 10% to 25% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid; from 35% to 49% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 35% to 50% by weight of the total composition of a nonionic surfactant. 153. The composition according to claim 139, comprising 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), and a medium-chain triglyceride oil comprises between about 50% to 60% caprylic acid and about 40% to 50% capric acid (MIGLYOL 812 N) in a weight ratio of about 2.5-3.5:3.5-4.5:2.5-3.5 or in a weight ratio of about 3:4:3. 154. The composition according to claim 139, further comprising an antioxidant. 155. The composition according to claim 139, comprising about 46 wt % of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, about 23 wt % of a medium-chain triglyceride oil comprising between about 50% to 60% caprylic acid and about 40% to 50% capric acid (MIGLYOL 812 N), about 31 wt % of surfactant chosen from polysorbate 20, polysorbate 80, polyoxyethylated castor oil with an HLB value of 2-14 (cremophor EL), or 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15), and about 150-300 ppm butylated hydroxy anisole. 156. The composition according to claim 139, comprising about 11%-40% by weight of the total composition of magnesium 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoat, and about 60-89% by weight of the total composition of a mixture comprising a polysorbate and a medium-chain triglyceride, wherein the weight ratio of the polysorbate and the medium-chain triglyceride is about 2:3 to 3:2. 157. The composition according to claim 156, further comprising from about 0%-15% by weight of the total composition of a co-solvent, wherein the co-solvent replaces from 0%-15% by weight of the mixture comprising a polysorbate and a medium-chain triglyceride. 158. The composition according to claim 139, wherein the particle size distribution of the emulsion is monodisperse. 159. The composition according to claim 139, wherein the composition is formulated for oral use. 160. The composition according to claim 139, wherein the composition is in the form of a capsule, liquid loaded tablet, or bead. 161. A self-nanoemulsifying drug delivery system (SNEDDS), self-microemulsifying drug delivery system (SMEDDS), or self-emulsifying drug delivery system (SEDDS) comprising a pharmaceutical composition comprising:
from 5% to 60% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof; from 15% to 60% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 10% to 60% by weight of the total composition of a nonionic surfactant. 162. The self-nanoemulsifying drug delivery system (SNEDDS), self-microemulsifying drug delivery system (SMEDDS), or self-emulsifying drug delivery system (SEDDS) according to claim 161, further comprising a co-solvent. 163. The self-nanoemulsifying drug delivery system (SNEDDS), self-microemulsifying drug delivery system (SMEDDS), or self-emulsifying drug delivery system (SEDDS) according to claim 161, comprising
2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, magnesium 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoate, calcium 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoate, or sodium 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoate. 164. A method for the prevention and/or treatment of a disease or condition chosen from:
a dyslipidemic condition; elevated triglyceride levels, non-HDL cholesterol levels, LDL cholesterol levels, and/or VLDL cholesterol levels; fatty liver disease; an inflammatory disease or condition; atherosclerosis; peripheral insulin resistance, a diabetic condition, and/or type 2 diabetes; in a subject in need thereof, comprising administering to the subject a pharmaceutically active amount of the composition according to claim 139. 165. The method according to claim 164, wherein the dyslipidemic condition is chosen from hypertriglyceridemia (HTG), dyslipidemia, and mixed dyslipidemia. 166. A method for the reduction of plasma insulin, blood glucose and/or serum triglycerides comprising administering to a human a pharmaceutically active amount of the composition according to claim 139. 167. A method for raising HDL cholesterol comprising administering to a human a pharmaceutically active amount of the composition according to claim 139. 168. The method according to claim 164, wherein the composition is administered once, twice, three, or four times per day. 169. The method according to claim 164, wherein the total dose of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, administered per day as a total dose ranges from about 50 mg to about 800 mg. 170. A method for the prevention and/or treatment of a disease or condition chosen from:
a dyslipidemic condition;
elevated triglyceride levels, non-HDL cholesterol levels, LDL cholesterol levels, and/or VLDL cholesterol levels;
fatty liver disease;
an inflammatory disease or condition;
atherosclerosis;
peripheral insulin resistance, a diabetic condition, and/or type 2 diabetes;
in a subject in need thereof, comprising administering to the subject a pharmaceutically active amount of a self-nanoemulsifying drug delivery system (SNEDDS), self-microemulsifying drug delivery system (SMEDDS), or self-emulsifying drug delivery system (SEDDS) according to claim 161. 171. A method for enhancing the hydrolysis, solubility, bioavailability, and/or absorption of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, comprising:
administering to a human in need thereof a composition comprising from 5% to 60% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, from 15% to 60% by weight of the total composition of a medium-chain triglyceride (MCT) oil, and from 10% to 60% by weight of the total composition of a nonionic surfactant, wherein the hydrolysis, solubility, bioavailability, and/or absorption of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof are enhanced when compared to 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof without a medium-chain triglyceride (MCT) oil or nonionic surfactant in the same amounts. | Compositions comprising 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof; a triglyceride; and a surfactant, as well as methods for their use. The present disclosure further relates to self-emulsifying drug delivery systems, such as SEDDS, SMEDDS, or SNEDDS comprising 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid.1-138. (canceled) 139. A composition comprising
from 5% to 60% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof; from 15% to 60% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 10% to 60% by weight of the total composition of a nonionic surfactant. 140. The composition according to claim 139, wherein the composition is a preconcentrate forming a nanoemulsion or microemulsion in contact with aqueous media. 141. The composition according to claim 139, further comprising a co-solvent. 142. The composition according to claim 139, wherein the 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof is present as its (R)-enantiomer, (S)-enantiomer, or a mixture of its (R)- and (S)-enantiomers. 143. The composition according to claim 139, comprising 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid
its (R)-enantiomer
its (S)-enantiomer
or
a mixture of its (R)- and (S)-enantiomer. 144. The composition according to claim 139, wherein the salt is formed from 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoate and an ion chosen from Li+, Na+, K+, NH4 +, Mg2+, Ca2+, and quaternized ions of meglumine, tris(hydroxymethyl)aminomethane, diethylamine, arginine, ethylenediamine, piperazine, and protonated chitosan salts. 145. The composition according to claim 139, wherein 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid is present as an ester derivative chosen from a C1-C6-alkyl or phenyl ester. 146. The composition according to claim 139, wherein the medium-chain triglyceride oil comprises C8-C10 triglycerides. 147. The composition according to claim 146, wherein the medium-chain triglyceride is derived from capric acid and caprylic acid. 148. The composition according to claim 146, wherein the medium-chain triglyceride oil comprises between 50% and 80% caprylic acid and between 20% to 50% capric acid or the medium-chain triglyceride oil comprises between about 50% to 60% caprylic acid and about 40% to 50% capric acid. 149. The composition according to claim 139, wherein the surfactant is chosen from polysorbates, sorbitan esters, ((poly(ethylene oxide))-(poly(propylene oxide)) (PEO-PPO)-block copolymers, polyoxyethylene (POE) castor oil, polyoxyethylene (POE) hydrogenated castor oil, polyoxyethylene (POE)-60-hydrogenated castor oil, polyoxyethylene (POE)-stearate, phospholipids, and mixtures thereof; from polyethylene (POE)-20-sorbitan monooleate, polyoxyethylene (20) sorbitan monooleate with a hydrophile-lipophile balance (HLB) value of 15.0 (TWEEN 80, CRILLET 4), polyoxyethylene (POE)-20-sorbitan monolaurate, polyoxyethylene sorbitan monostearate with an HLB value of 14.9 (TWEEN 60), polyoxyethylene (20) sorbitan monopalmitate with an HLB value of 15.6 (TWEEN 40), pharmaceutical grade polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20, CRILLET 1), Poloxamer 188 (PLURONIC F68, LUTROL F68), polyoxyethylene (POE) alkyl ethers, polyoxyethylene (POE)-10-oleyl ether, and polyethylene glycol oleyl ether with an HLB value of 12.4 (BRIJ 96 V), and mixtures thereof; from 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15), polyoxyethylated castor oil with a hydrophile-lipophile balance (HLB) value of 2-14 (CREMOPHOR EL), hydrogenated polyoxyethylated castor oil with an HLB value of 14-16 (CREMOPHOR RH40), polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), polyoxyethylene (20) sorbitan monopalmitate with an HLB value of 15.6 (TWEEN 40), polyoxyethylene sorbitan monostearate with an HLB value of 14.9 (TWEEN 60), polyoxyethylene (20) sorbitan monooleate with an HLB value of 15.0 (TWEEN 80), sorbitan monooleate with an HLB value of 4.3 (SPAN 80), polyoxyethylene (10) oleyl ether with an HLB value of 12 (BRIJ 97), and mixtures thereof. 150. The composition according to claim 141, comprising from 0% to 20% or from 0% to 10% by weight of the total composition of co-solvent and wherein the co-solvent is chosen from C1 to C6 alcohols, ethanol, benzyl alcohol, alkane diols and triols, glycol ethers, propylene glycol (PG), glycerol, tetraglycol, polyethylene glycols (PEG), PEG 400, pyrrolidine derivatives, 2-pyrrolidone, triacetin, and mixtures thereof. 151. The composition according to claim 139, comprising:
2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, an MCT oil, and a surfactant chosen from polysorbates, and a co-solvent PEG 400; 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, an MCT oil, polyoxyethylene (20) sorbitan monooleate with an HLB value of 15.0 (TWEEN 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15), and PEG 400; 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, an MCT oil, polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), polyoxyethylene (20) sorbitan monooleate with an HLB value of 15.0 (TWEEN 80), and 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15); 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, a MCT oil, polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15), and PEG 400; 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, a MCT oil, and polyoxyethylene (20) sorbitan monooleate with an HLB value of 15.0 (TWEEN 80); 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, a MCT oil, and polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20); or 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), and a medium-chain triglyceride oil comprising between about 50% to 60% caprylic acid and about 40% to 50% capric acid (MIGLYOL 812 N). 152. The composition according to claim 139, comprising:
from 5% to 46% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid; from 20% to 49% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 30% to 60% by weight of the total composition of a nonionic surfactant; from 15% to 48% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid; from 20% to 40% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 30% to 50% by weight of the total composition of a nonionic surfactant; from 40% to 48% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid; from 20% to 30% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 30% to 40% by weight of the total composition of a nonionic surfactant; or from 10% to 25% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid; from 35% to 49% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 35% to 50% by weight of the total composition of a nonionic surfactant. 153. The composition according to claim 139, comprising 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, polyoxyethylene (20) sorbitan monolaurate with an HLB value of 16.0 (TWEEN 20), and a medium-chain triglyceride oil comprises between about 50% to 60% caprylic acid and about 40% to 50% capric acid (MIGLYOL 812 N) in a weight ratio of about 2.5-3.5:3.5-4.5:2.5-3.5 or in a weight ratio of about 3:4:3. 154. The composition according to claim 139, further comprising an antioxidant. 155. The composition according to claim 139, comprising about 46 wt % of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, about 23 wt % of a medium-chain triglyceride oil comprising between about 50% to 60% caprylic acid and about 40% to 50% capric acid (MIGLYOL 812 N), about 31 wt % of surfactant chosen from polysorbate 20, polysorbate 80, polyoxyethylated castor oil with an HLB value of 2-14 (cremophor EL), or 2-hydroxyethyl 12-hydroxyoctadecanoate (SOLUTOL HS15), and about 150-300 ppm butylated hydroxy anisole. 156. The composition according to claim 139, comprising about 11%-40% by weight of the total composition of magnesium 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoat, and about 60-89% by weight of the total composition of a mixture comprising a polysorbate and a medium-chain triglyceride, wherein the weight ratio of the polysorbate and the medium-chain triglyceride is about 2:3 to 3:2. 157. The composition according to claim 156, further comprising from about 0%-15% by weight of the total composition of a co-solvent, wherein the co-solvent replaces from 0%-15% by weight of the mixture comprising a polysorbate and a medium-chain triglyceride. 158. The composition according to claim 139, wherein the particle size distribution of the emulsion is monodisperse. 159. The composition according to claim 139, wherein the composition is formulated for oral use. 160. The composition according to claim 139, wherein the composition is in the form of a capsule, liquid loaded tablet, or bead. 161. A self-nanoemulsifying drug delivery system (SNEDDS), self-microemulsifying drug delivery system (SMEDDS), or self-emulsifying drug delivery system (SEDDS) comprising a pharmaceutical composition comprising:
from 5% to 60% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof; from 15% to 60% by weight of the total composition of a medium-chain triglyceride (MCT) oil; and from 10% to 60% by weight of the total composition of a nonionic surfactant. 162. The self-nanoemulsifying drug delivery system (SNEDDS), self-microemulsifying drug delivery system (SMEDDS), or self-emulsifying drug delivery system (SEDDS) according to claim 161, further comprising a co-solvent. 163. The self-nanoemulsifying drug delivery system (SNEDDS), self-microemulsifying drug delivery system (SMEDDS), or self-emulsifying drug delivery system (SEDDS) according to claim 161, comprising
2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, magnesium 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoate, calcium 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoate, or sodium 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoate. 164. A method for the prevention and/or treatment of a disease or condition chosen from:
a dyslipidemic condition; elevated triglyceride levels, non-HDL cholesterol levels, LDL cholesterol levels, and/or VLDL cholesterol levels; fatty liver disease; an inflammatory disease or condition; atherosclerosis; peripheral insulin resistance, a diabetic condition, and/or type 2 diabetes; in a subject in need thereof, comprising administering to the subject a pharmaceutically active amount of the composition according to claim 139. 165. The method according to claim 164, wherein the dyslipidemic condition is chosen from hypertriglyceridemia (HTG), dyslipidemia, and mixed dyslipidemia. 166. A method for the reduction of plasma insulin, blood glucose and/or serum triglycerides comprising administering to a human a pharmaceutically active amount of the composition according to claim 139. 167. A method for raising HDL cholesterol comprising administering to a human a pharmaceutically active amount of the composition according to claim 139. 168. The method according to claim 164, wherein the composition is administered once, twice, three, or four times per day. 169. The method according to claim 164, wherein the total dose of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, administered per day as a total dose ranges from about 50 mg to about 800 mg. 170. A method for the prevention and/or treatment of a disease or condition chosen from:
a dyslipidemic condition;
elevated triglyceride levels, non-HDL cholesterol levels, LDL cholesterol levels, and/or VLDL cholesterol levels;
fatty liver disease;
an inflammatory disease or condition;
atherosclerosis;
peripheral insulin resistance, a diabetic condition, and/or type 2 diabetes;
in a subject in need thereof, comprising administering to the subject a pharmaceutically active amount of a self-nanoemulsifying drug delivery system (SNEDDS), self-microemulsifying drug delivery system (SMEDDS), or self-emulsifying drug delivery system (SEDDS) according to claim 161. 171. A method for enhancing the hydrolysis, solubility, bioavailability, and/or absorption of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, comprising:
administering to a human in need thereof a composition comprising from 5% to 60% by weight of the total composition of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof, from 15% to 60% by weight of the total composition of a medium-chain triglyceride (MCT) oil, and from 10% to 60% by weight of the total composition of a nonionic surfactant, wherein the hydrolysis, solubility, bioavailability, and/or absorption of 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof are enhanced when compared to 2-((5Z,8Z,11Z,14Z,17Z)-icosa-5,8,11,14,17-pentaenyloxy)butanoic acid, an ester derivative, or a pharmaceutically acceptable salt thereof without a medium-chain triglyceride (MCT) oil or nonionic surfactant in the same amounts. | 1,600 |
1,006 | 16,027,997 | 1,617 | A synergistic agricultural formula including at least one diacyl or diaryl urea, such as a N,N′-diformylurea, and at least one plant growth regulator. This synergistic agricultural formula gives those skilled in the art the ability to regulate important phenotypical parameters that lead to a variety of important agronomic and horticulture traits which improve crop yield parameters leading beyond that of its individual components. | 1. An agricultural formula comprising 30 to 0.1% of at least one diacyl or diaryl urea and 0.001 to 99.9 wt % of at least one plant growth regulator. 2. The agricultural formula of claim 1, wherein said at least one diacyl or diaryl urea is only at least one diacyl urea. 3. The agricultural formula of claim 1, wherein said at least one diacyl or diaryl urea is only diformyl urea. 4. The agricultural formula of claim 1, wherein said formula only includes plant growth regulators selected from the group consisting of ethylene, auxins, cytokinins, gibberellins, abscisic acid, brassinosteroids, jasmonates, salicylic acids, peptides, polyamines, nitric oxide, strigolactones, precursors, derivatives and mixtures thereof. 5. The agricultural formula of claim 4, wherein said formula only includes plant growth regulators selected from the group consisting of auxins, cytokinins, and gibberellins. 6. The agricultural formula of claim 5, wherein said formula only includes plant growth regulators selected from the group consisting of cytokinins and gibberellins. 7. The agricultural formula of claims 5, wherein said formula only includes plant growth regulators selected from the group consisting of cytokinins and auxins. 8. The agricultural formula of claim 1, wherein said at least one diacyl or diaryl urea and said at least one plant growth regulator are the only agriculturally active ingredients. 9. The agricultural formula of claim 1 comprising 20 to 0.1 wt. % of at least one diacyl or diaryl urea and 0.001 to 5 wt. % of at least one plant growth regulator. 10. The agricultural formula of claim 1 comprising 20 to 0.1 wt. % of at least one diacyl or diaryl urea and 0.001 to 1 wt. % of at least one plant growth regulator. 11. The agricultural formula of claim 1 comprising 20 to 0.1 wt. % of at least one diacyl or diaryl urea and 0.001 to 0.1 wt. % of at least one plant growth regulator. 12. The agricultural formula of claim 1 comprising 15 to 10 wt. % of at least one diacyl or diaryl urea and 0.001 to 1 wt. % of at least one plant growth regulator. 13. The agricultural formula of claim 1, wherein said formula only includes plant growth regulators selected from the group consisting of indole-3-butyric acid, indole-3-acetic acid, gibberellic acid, and kinetin. 14. The agricultural formula of claim 13, wherein said formula includes 0.2-0.0005 wt. % kinetin. 15. The agricultural formula of claim 13, wherein said formula includes 0.1-0.0003 wt. % gibberellic acid. 16. The agricultural formula of claim 13, wherein said formula includes 0.1-0.0003 wt. % indole-3-butyric acid or indole-3-acetic acid. 17. The agricultural formula of claim 13, wherein said formula includes 0.2-0.0005 wt. % kinetin, 0.1-0.0003 wt. % gibberellic acid, and 0.1-0.0003 wt. % indole-3-butyric acid or indole-3-acetic acid. 18. An agricultural formula consisting essentially of,
at least one diacyl or diaryl urea, at least one plant growth regulator, water, and less than 5 wt. % oil and surfactants, wherein said at least one diacyl or diaryl urea and said at least one plant growth regulator are the only agriculturally active ingredients. 19. The agricultural formula of claim 18, wherein said at least one diacyl or diaryl urea is only diformylurea. 20. The agricultural formula of claim 19, wherein said formula only includes plant growth regulators selected from the group consisting of indole-3-butyric acid, indole-3-acetic acid, gibberellic acid, and kinetin. | A synergistic agricultural formula including at least one diacyl or diaryl urea, such as a N,N′-diformylurea, and at least one plant growth regulator. This synergistic agricultural formula gives those skilled in the art the ability to regulate important phenotypical parameters that lead to a variety of important agronomic and horticulture traits which improve crop yield parameters leading beyond that of its individual components.1. An agricultural formula comprising 30 to 0.1% of at least one diacyl or diaryl urea and 0.001 to 99.9 wt % of at least one plant growth regulator. 2. The agricultural formula of claim 1, wherein said at least one diacyl or diaryl urea is only at least one diacyl urea. 3. The agricultural formula of claim 1, wherein said at least one diacyl or diaryl urea is only diformyl urea. 4. The agricultural formula of claim 1, wherein said formula only includes plant growth regulators selected from the group consisting of ethylene, auxins, cytokinins, gibberellins, abscisic acid, brassinosteroids, jasmonates, salicylic acids, peptides, polyamines, nitric oxide, strigolactones, precursors, derivatives and mixtures thereof. 5. The agricultural formula of claim 4, wherein said formula only includes plant growth regulators selected from the group consisting of auxins, cytokinins, and gibberellins. 6. The agricultural formula of claim 5, wherein said formula only includes plant growth regulators selected from the group consisting of cytokinins and gibberellins. 7. The agricultural formula of claims 5, wherein said formula only includes plant growth regulators selected from the group consisting of cytokinins and auxins. 8. The agricultural formula of claim 1, wherein said at least one diacyl or diaryl urea and said at least one plant growth regulator are the only agriculturally active ingredients. 9. The agricultural formula of claim 1 comprising 20 to 0.1 wt. % of at least one diacyl or diaryl urea and 0.001 to 5 wt. % of at least one plant growth regulator. 10. The agricultural formula of claim 1 comprising 20 to 0.1 wt. % of at least one diacyl or diaryl urea and 0.001 to 1 wt. % of at least one plant growth regulator. 11. The agricultural formula of claim 1 comprising 20 to 0.1 wt. % of at least one diacyl or diaryl urea and 0.001 to 0.1 wt. % of at least one plant growth regulator. 12. The agricultural formula of claim 1 comprising 15 to 10 wt. % of at least one diacyl or diaryl urea and 0.001 to 1 wt. % of at least one plant growth regulator. 13. The agricultural formula of claim 1, wherein said formula only includes plant growth regulators selected from the group consisting of indole-3-butyric acid, indole-3-acetic acid, gibberellic acid, and kinetin. 14. The agricultural formula of claim 13, wherein said formula includes 0.2-0.0005 wt. % kinetin. 15. The agricultural formula of claim 13, wherein said formula includes 0.1-0.0003 wt. % gibberellic acid. 16. The agricultural formula of claim 13, wherein said formula includes 0.1-0.0003 wt. % indole-3-butyric acid or indole-3-acetic acid. 17. The agricultural formula of claim 13, wherein said formula includes 0.2-0.0005 wt. % kinetin, 0.1-0.0003 wt. % gibberellic acid, and 0.1-0.0003 wt. % indole-3-butyric acid or indole-3-acetic acid. 18. An agricultural formula consisting essentially of,
at least one diacyl or diaryl urea, at least one plant growth regulator, water, and less than 5 wt. % oil and surfactants, wherein said at least one diacyl or diaryl urea and said at least one plant growth regulator are the only agriculturally active ingredients. 19. The agricultural formula of claim 18, wherein said at least one diacyl or diaryl urea is only diformylurea. 20. The agricultural formula of claim 19, wherein said formula only includes plant growth regulators selected from the group consisting of indole-3-butyric acid, indole-3-acetic acid, gibberellic acid, and kinetin. | 1,600 |
1,007 | 16,535,704 | 1,628 | Provided herein are methods for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is also being administered an organic anion transporter-1 (OAT1) substrate. The method comprises administering to the subject a therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, and monitoring the subject for signs and symptoms of toxicity and clinical response associated with the OAT1 substrate. | 1. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is being administered an organic anion transporter-1 (OAT1) substrate to treat a disease or disorder, the method comprising:
discontinuing administration of the OAT1 substrate, prior to administering said dichlorphenamide, or a pharmaceutically acceptable salt thereof, and initiating administration to the subject of a therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered to the subject to treat a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants. 2. (canceled) 3. The method of claim 1, further comprising informing the subject or a medical care worker that co-administration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, and the OAT1 substrate may result in increased exposure of the OAT1 substrate. 4. The method of claim 1, further comprising informing the subject or a medical care worker that co-administration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, and the OAT1 substrate may result in increased risk of one or more exposure-related adverse reactions associated with the OAT1 substrate. 5.-22. (canceled) 23. The method of claim 1, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is between 25 mg and 200 mg per day. 24. The method of claim 1, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg twice daily. 25.-30. (canceled) 31. The method of claim 1, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg once daily. 32. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, the method comprising:
administering to the subject between 25 mg and 200 mg per day of dichlorphenamide, or a pharmaceutically acceptable salt thereof, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof is administered in the absence of an OAT1 substrate, and wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered to the subject to treat a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants. 33. The method of claim 32, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg twice daily. 34. The method of claim 32, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg once daily. | Provided herein are methods for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is also being administered an organic anion transporter-1 (OAT1) substrate. The method comprises administering to the subject a therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, and monitoring the subject for signs and symptoms of toxicity and clinical response associated with the OAT1 substrate.1. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is being administered an organic anion transporter-1 (OAT1) substrate to treat a disease or disorder, the method comprising:
discontinuing administration of the OAT1 substrate, prior to administering said dichlorphenamide, or a pharmaceutically acceptable salt thereof, and initiating administration to the subject of a therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered to the subject to treat a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants. 2. (canceled) 3. The method of claim 1, further comprising informing the subject or a medical care worker that co-administration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, and the OAT1 substrate may result in increased exposure of the OAT1 substrate. 4. The method of claim 1, further comprising informing the subject or a medical care worker that co-administration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, and the OAT1 substrate may result in increased risk of one or more exposure-related adverse reactions associated with the OAT1 substrate. 5.-22. (canceled) 23. The method of claim 1, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is between 25 mg and 200 mg per day. 24. The method of claim 1, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg twice daily. 25.-30. (canceled) 31. The method of claim 1, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg once daily. 32. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, the method comprising:
administering to the subject between 25 mg and 200 mg per day of dichlorphenamide, or a pharmaceutically acceptable salt thereof, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof is administered in the absence of an OAT1 substrate, and wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered to the subject to treat a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants. 33. The method of claim 32, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg twice daily. 34. The method of claim 32, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg once daily. | 1,600 |
1,008 | 16,253,515 | 1,628 | Provided herein is a method of administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is also being administered an organic anion transporter-1 (OAT1) inhibitor and/or an organic anion transporter-3 (OAT3) inhibitor. The method comprises administering to the subject a therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, monitoring the subject for signs and/or symptoms of toxicity associated with the dichlorphenamide, or a pharmaceutically acceptable salt thereof, and adjusting the therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, when the subject is experiencing a sign and/or symptom of toxicity associated with the dichlorphenamide, or a pharmaceutically acceptable salt thereof. | 1.-29. (canceled) 30. A method of treating a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants in a subject in need thereof,
wherein the subject is exhibiting one or more signs of dichlorphenamide toxicity as a result of increase in exposure of dichlorphenamide caused by the subject being co-administered:
(i) an organic anion transporter-1 (OAT1) substrate dichlorphenamide, or a pharmaceutically acceptable salt thereof, at an initial dosage of up to 200 mg daily to treat a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants, and
(ii) an organic anion transporter-1 (OAT1) inhibitor to treat a disease other than primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants, wherein the OAT1 inhibitor is chosen from furosemide, diclofenac, naproxen, bumetanide, captopril, candesartan, losartan, chlorothiazide, cimetidine, ranitidine, telmisartan, olmesartan, simvastatin, fluvastatin, cefaclor, methotrexate, cefadroxil, cefoperazone, ceftizoxime, piperacillin, tazobactam, sulbactam, zidovudine, adefovir, cidofovir, ketorolac, diflunisal, and any combination thereof,
wherein the one or more signs of dichlorphenamide toxicity is chosen from paresthesia, cognitive disorder, dysgeusia, confusional state, hypersensitivity reactions, anaphylaxis reactions, hypokalemia, metabolic acidosis, falls, amnesia, cardiac failure, condition aggravated, convulsion, fetal death, hallucination, nephrolithiasis, pancytopenia, psychotic disorder, renal tubular necrosis, stupor, syncope, and tremor,
the method comprising:
administering a reduced dose of dichlorphenamide, or a pharmaceutically acceptable salt thereof, once daily to the subject, wherein the reduced dose is less than the initial dose; and
continuing administration of the OAT1 inhibitor. 31. A method of treating a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants in a subject in need thereof,
wherein the subject is also in need of treatment with an organic anion transporter-1 (OAT1) inhibitor to treat a disease other than primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants, and wherein the OAT1 inhibitor is chosen from furosemide, diclofenac, naproxen, bumetanide, captopril, candesartan, losartan, chlorothiazide, cimetidine, ranitidine, telmisartan, olmesartan, simvastatin, fluvastatin, cefaclor, methotrexate, cefadroxil, cefoperazone, ceftizoxime, piperacillin, tazobactam, sulbactam, zidovudine, adefovir, cidofovir, ketorolac, diflunisal, and any combination thereof,
the method comprising:
administering the OAT1 inhibitor to the subject; and
subsequently administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to the subject at a reduced dose to compensate for the expected increase in exposure resulting from co-administration of the OAT1 inhibitor and the OAT1 substrate dichlorphenamide, or a pharmaceutically acceptable salt thereof,
wherein the reduced dose is relative to what the subject would be administered if the subject was not being administered the OAT1 inhibitor. | Provided herein is a method of administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is also being administered an organic anion transporter-1 (OAT1) inhibitor and/or an organic anion transporter-3 (OAT3) inhibitor. The method comprises administering to the subject a therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, monitoring the subject for signs and/or symptoms of toxicity associated with the dichlorphenamide, or a pharmaceutically acceptable salt thereof, and adjusting the therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, when the subject is experiencing a sign and/or symptom of toxicity associated with the dichlorphenamide, or a pharmaceutically acceptable salt thereof.1.-29. (canceled) 30. A method of treating a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants in a subject in need thereof,
wherein the subject is exhibiting one or more signs of dichlorphenamide toxicity as a result of increase in exposure of dichlorphenamide caused by the subject being co-administered:
(i) an organic anion transporter-1 (OAT1) substrate dichlorphenamide, or a pharmaceutically acceptable salt thereof, at an initial dosage of up to 200 mg daily to treat a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants, and
(ii) an organic anion transporter-1 (OAT1) inhibitor to treat a disease other than primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants, wherein the OAT1 inhibitor is chosen from furosemide, diclofenac, naproxen, bumetanide, captopril, candesartan, losartan, chlorothiazide, cimetidine, ranitidine, telmisartan, olmesartan, simvastatin, fluvastatin, cefaclor, methotrexate, cefadroxil, cefoperazone, ceftizoxime, piperacillin, tazobactam, sulbactam, zidovudine, adefovir, cidofovir, ketorolac, diflunisal, and any combination thereof,
wherein the one or more signs of dichlorphenamide toxicity is chosen from paresthesia, cognitive disorder, dysgeusia, confusional state, hypersensitivity reactions, anaphylaxis reactions, hypokalemia, metabolic acidosis, falls, amnesia, cardiac failure, condition aggravated, convulsion, fetal death, hallucination, nephrolithiasis, pancytopenia, psychotic disorder, renal tubular necrosis, stupor, syncope, and tremor,
the method comprising:
administering a reduced dose of dichlorphenamide, or a pharmaceutically acceptable salt thereof, once daily to the subject, wherein the reduced dose is less than the initial dose; and
continuing administration of the OAT1 inhibitor. 31. A method of treating a disease chosen from primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants in a subject in need thereof,
wherein the subject is also in need of treatment with an organic anion transporter-1 (OAT1) inhibitor to treat a disease other than primary hyperkalemic periodic paralysis, primary hypokalemic periodic paralysis, and related variants, and wherein the OAT1 inhibitor is chosen from furosemide, diclofenac, naproxen, bumetanide, captopril, candesartan, losartan, chlorothiazide, cimetidine, ranitidine, telmisartan, olmesartan, simvastatin, fluvastatin, cefaclor, methotrexate, cefadroxil, cefoperazone, ceftizoxime, piperacillin, tazobactam, sulbactam, zidovudine, adefovir, cidofovir, ketorolac, diflunisal, and any combination thereof,
the method comprising:
administering the OAT1 inhibitor to the subject; and
subsequently administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to the subject at a reduced dose to compensate for the expected increase in exposure resulting from co-administration of the OAT1 inhibitor and the OAT1 substrate dichlorphenamide, or a pharmaceutically acceptable salt thereof,
wherein the reduced dose is relative to what the subject would be administered if the subject was not being administered the OAT1 inhibitor. | 1,600 |
1,009 | 15,954,291 | 1,632 | Methods are disclosed for reprogramming a somatic cell, including an adherent cell and a cell in suspension, into an induced pluripotent stem comprising expressing exogenous Sox-2, exogenous Klf-4, exogenous Oct3/4 from DNA that has not integrated into the genome of the somatic cell, suppressing p53 activity within the somatic cell, and exposing the somatic cell to reprogramming-assistance factors comprising an exogenous Alk-5 inhibitor, an exogenous histone deacetylase inhibitor, and an exogenous activator of glycolysis. Compositions and kits for use in such methods are also disclosed as are cells made by such a method. | 1. A method for reprogramming a somatic cell into an induced pluripotent stem (iPS) cell in vitro comprising:
expressing exogenous sex determining region Y-box 2 (Sox-2), Kruppel-like factor 4 (Klf-4), and octamer-binding transcription factor 3/4 (Oct3/4) in the somatic cell from DNA that has not integrated into genomic DNA of the somatic cell; inhibiting p53 activity in the somatic cell; and culturing the somatic cell in a reprogramming medium comprising an exogenous activating receptor-like kinase 5 (Alk-5) inhibitor, an exogenous histone deacetylase inhibitor, and an exogenous activator of glycolysis to obtain an iPS cell. 2. The method according to claim 1, wherein the method further comprises expressing exogenous Epstein-Bar nuclear antigen-1 (EBNA-1) in the somatic cell from DNA that has not integrated into the genomic DNA of the somatic cell, and wherein the DNA that has not integrated into the genomic DNA of the somatic cell comprises at least one plasmid with an Epstein-Barr virus origin of replication (oriP). 3. The method according to claim 1, wherein one or more of lung myelocytomatosis oncogene (L-Myc), c-myelocytomatosis oncogene (c-Myc), Lin28, simian virus 40 large T antigen, and Nanog are not exogenously expressed. 4. The method according to claim 1, further comprising maintaining the cell in a dedifferentiation maintenance medium comprising basic fibroblast growth factor and transforming growth factor beta after being cultured in the reprogramming medium. 5. The method according to claim 1, wherein the culturing does not require feeder cells. 6. The method according to claim 3, wherein the reprogramming efficiency exceeds 0.0006%. 7. The method according to claim 1, wherein inhibiting p53 activity in the somatic cell inhibits p53-induced cell cycle arrest or p53-induced apoptosis. 8. The method according to claim 1, wherein inhibiting p53 activity in the somatic cell comprises suppressing p53 expression in the somatic cell. 9. The method according to claim 8, wherein suppressing p53 expression comprises expressing antisense p53 RNA in the somatic cell from DNA that has not integrated into the genomic DNA of the somatic cell. 10. The method according to claim 9, wherein the Alk-5 inhibitor comprises 3-(6-Methyl-2-pyridinyl)-N-phenyl-4-(4-quinolinyl)-1H-pyrazole-1-carbothioamide (A83-01), the histone deacetylase inhibitor comprises sodium butyrate or valproic acid, and the activator of glycolysis comprises a phosphoinositide-dependent protein kinase-1 inhibitor selected from 5-(4-Chloro-phenyl)-3-phenyl-pent-2-enoic acid (PS48); α,α,-Dimethyl-4-[2-methyl-8-[2-(3-pyridinyl)ethynyl]-1H-imidazo[4,5-c]quinolin-1-yl]-benzeneacetonitrile (BAG956); N-[3-[[5-Iodo-4-[[3-[(2-thienylcarbonyl)amino]propyl]amino]-2-pyrimidinyl]amino]phenyl]-1-pyrrolidinecarboxamide (BX795); (3S,6R)-1-[6-(3-Amino-1H-indazol-6-yl)-2-(methylamino)-4-pyrimidinyl]-N-cyclohexyl-6-methyl-3-piperidinecarboxamide (GSK 2334470); 2-Amino-N-[4-[5-(2-phenanthrenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]phenyl]acetamide (OSU03012); and 4-Dodecyl-N-1,3,4-thiadiazol-2-yl-benzenesulfonamide (PHT427). 11. The method according to claim 1, wherein the somatic cell is an isolated cord blood or peripheral blood mononuclear cell, and wherein the method further comprises pre-culturing the isolated cord blood or peripheral blood mononuclear cell in hematopoietic stem cell expansion media. 12. The method according to claim 2, which yields an integration-free, virus-free, exogenous oncogene-free iPS cell. 13. The method according to claim 12, wherein the iPS cell is a human iPS cell. 14. The method according to claim 12, wherein the iPS cell is differentiated into an endodermal, mesodermal, or ectodermal cell. 15. A method for reprogramming a somatic cell into an iPS cell in vitro comprising:
expressing exogenous Sox-2, Klf-4, and Oct-3/4 in the somatic cell from episomal DNA; suppressing p53 expression in the somatic cell; and culturing the somatic cell in a reprogramming medium comprising at least three different exogenous reprogramming-assistance factors, the exogenous reprogramming-assistance factors being at least an Alk-5 inhibitor, a histone deacetylase inhibitor, and an activator of glycolysis, to obtain an iPS cell free of exogenous oncogenes and exogenous viral elements. 16. (canceled) 17. The method according to claim 15, further comprising expressing EBNA-1 in the somatic cell and wherein Sox-2, Klf-4, Oct-3/4, and EBNA-1 are expressed from at least one plasmid with an Epstein-Barr virus origin of replication (oriP). 18. The method according to claim 15, wherein suppressing p53 expression in the somatic cell comprises expressing antisense p53 RNA in the somatic cell. 19. (canceled) 20. (canceled) 21. The method according to claim 1, the somatic cell being free of exogenous oncogenes and retroviruses. 22. The method according to claim 3, wherein the reprogramming efficiency exceeds 0.001%. 23. The method according to claim 1, wherein colonies of iPS cells are obtained and 100% of the colonies express stage-specific embryonic antigen-4 (SSEA-4). 24. The method according to claim 1, wherein colonies of iPS cells are obtained, the colonies expressing SSEA-4, Nanog, Oct3/4, and TRA160. 25. The method according to claim 1, the somatic cell being an adherent cell. | Methods are disclosed for reprogramming a somatic cell, including an adherent cell and a cell in suspension, into an induced pluripotent stem comprising expressing exogenous Sox-2, exogenous Klf-4, exogenous Oct3/4 from DNA that has not integrated into the genome of the somatic cell, suppressing p53 activity within the somatic cell, and exposing the somatic cell to reprogramming-assistance factors comprising an exogenous Alk-5 inhibitor, an exogenous histone deacetylase inhibitor, and an exogenous activator of glycolysis. Compositions and kits for use in such methods are also disclosed as are cells made by such a method.1. A method for reprogramming a somatic cell into an induced pluripotent stem (iPS) cell in vitro comprising:
expressing exogenous sex determining region Y-box 2 (Sox-2), Kruppel-like factor 4 (Klf-4), and octamer-binding transcription factor 3/4 (Oct3/4) in the somatic cell from DNA that has not integrated into genomic DNA of the somatic cell; inhibiting p53 activity in the somatic cell; and culturing the somatic cell in a reprogramming medium comprising an exogenous activating receptor-like kinase 5 (Alk-5) inhibitor, an exogenous histone deacetylase inhibitor, and an exogenous activator of glycolysis to obtain an iPS cell. 2. The method according to claim 1, wherein the method further comprises expressing exogenous Epstein-Bar nuclear antigen-1 (EBNA-1) in the somatic cell from DNA that has not integrated into the genomic DNA of the somatic cell, and wherein the DNA that has not integrated into the genomic DNA of the somatic cell comprises at least one plasmid with an Epstein-Barr virus origin of replication (oriP). 3. The method according to claim 1, wherein one or more of lung myelocytomatosis oncogene (L-Myc), c-myelocytomatosis oncogene (c-Myc), Lin28, simian virus 40 large T antigen, and Nanog are not exogenously expressed. 4. The method according to claim 1, further comprising maintaining the cell in a dedifferentiation maintenance medium comprising basic fibroblast growth factor and transforming growth factor beta after being cultured in the reprogramming medium. 5. The method according to claim 1, wherein the culturing does not require feeder cells. 6. The method according to claim 3, wherein the reprogramming efficiency exceeds 0.0006%. 7. The method according to claim 1, wherein inhibiting p53 activity in the somatic cell inhibits p53-induced cell cycle arrest or p53-induced apoptosis. 8. The method according to claim 1, wherein inhibiting p53 activity in the somatic cell comprises suppressing p53 expression in the somatic cell. 9. The method according to claim 8, wherein suppressing p53 expression comprises expressing antisense p53 RNA in the somatic cell from DNA that has not integrated into the genomic DNA of the somatic cell. 10. The method according to claim 9, wherein the Alk-5 inhibitor comprises 3-(6-Methyl-2-pyridinyl)-N-phenyl-4-(4-quinolinyl)-1H-pyrazole-1-carbothioamide (A83-01), the histone deacetylase inhibitor comprises sodium butyrate or valproic acid, and the activator of glycolysis comprises a phosphoinositide-dependent protein kinase-1 inhibitor selected from 5-(4-Chloro-phenyl)-3-phenyl-pent-2-enoic acid (PS48); α,α,-Dimethyl-4-[2-methyl-8-[2-(3-pyridinyl)ethynyl]-1H-imidazo[4,5-c]quinolin-1-yl]-benzeneacetonitrile (BAG956); N-[3-[[5-Iodo-4-[[3-[(2-thienylcarbonyl)amino]propyl]amino]-2-pyrimidinyl]amino]phenyl]-1-pyrrolidinecarboxamide (BX795); (3S,6R)-1-[6-(3-Amino-1H-indazol-6-yl)-2-(methylamino)-4-pyrimidinyl]-N-cyclohexyl-6-methyl-3-piperidinecarboxamide (GSK 2334470); 2-Amino-N-[4-[5-(2-phenanthrenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]phenyl]acetamide (OSU03012); and 4-Dodecyl-N-1,3,4-thiadiazol-2-yl-benzenesulfonamide (PHT427). 11. The method according to claim 1, wherein the somatic cell is an isolated cord blood or peripheral blood mononuclear cell, and wherein the method further comprises pre-culturing the isolated cord blood or peripheral blood mononuclear cell in hematopoietic stem cell expansion media. 12. The method according to claim 2, which yields an integration-free, virus-free, exogenous oncogene-free iPS cell. 13. The method according to claim 12, wherein the iPS cell is a human iPS cell. 14. The method according to claim 12, wherein the iPS cell is differentiated into an endodermal, mesodermal, or ectodermal cell. 15. A method for reprogramming a somatic cell into an iPS cell in vitro comprising:
expressing exogenous Sox-2, Klf-4, and Oct-3/4 in the somatic cell from episomal DNA; suppressing p53 expression in the somatic cell; and culturing the somatic cell in a reprogramming medium comprising at least three different exogenous reprogramming-assistance factors, the exogenous reprogramming-assistance factors being at least an Alk-5 inhibitor, a histone deacetylase inhibitor, and an activator of glycolysis, to obtain an iPS cell free of exogenous oncogenes and exogenous viral elements. 16. (canceled) 17. The method according to claim 15, further comprising expressing EBNA-1 in the somatic cell and wherein Sox-2, Klf-4, Oct-3/4, and EBNA-1 are expressed from at least one plasmid with an Epstein-Barr virus origin of replication (oriP). 18. The method according to claim 15, wherein suppressing p53 expression in the somatic cell comprises expressing antisense p53 RNA in the somatic cell. 19. (canceled) 20. (canceled) 21. The method according to claim 1, the somatic cell being free of exogenous oncogenes and retroviruses. 22. The method according to claim 3, wherein the reprogramming efficiency exceeds 0.001%. 23. The method according to claim 1, wherein colonies of iPS cells are obtained and 100% of the colonies express stage-specific embryonic antigen-4 (SSEA-4). 24. The method according to claim 1, wherein colonies of iPS cells are obtained, the colonies expressing SSEA-4, Nanog, Oct3/4, and TRA160. 25. The method according to claim 1, the somatic cell being an adherent cell. | 1,600 |
1,010 | 14,206,369 | 1,619 | Described are transdermal drug delivery compositions comprising amphetamine, methods of making them and therapeutic methods using them. The compositions are provided in a flexible, finite form (e.g. “patch”-type systems) and comprise a polymer matrix that includes amphetamine and an acrylic block copolymer. | 1. A composition for the transdermal delivery of amphetamine in the form of a flexible finite system for topical application, comprising a polymer matrix comprising amphetamine or a pharmaceutically acceptable salt or prodrug thereof, wherein the polymer matrix comprises an acrylic block copolymer. 2. The composition of claim 1, wherein the acrylic block copolymer is comprised of one or more monomers selected from the group consisting of methyl acrylate, methyl methacrylate, butyl acrylate, butyl methacrylate, hexyl acrylate, hexyl methacrylate, 2-ethylbutyl acrylate, 2-ethylbutyl methacrylate, isooctyl acrylate, isooctyl methacrylate, 2-ethylhexyl acrylate, 2-ethylhexyl methacrylate, decyl acrylate, decyl methacrylate, dodecyl acrylate, dodecyl methacrylate, tridecyl acrylate, tridecyl methacrylate, and octyl acrylamide. 2. The composition of claim 1, wherein the acrylic block copolymer comprises one or more of poly(butyl acrylate), poly(methyl methacrylate), methyl methacrylate/butyl acrylate (MMA/BA) blocks, and methyl methacrylate/butyl acrylate/alpha methyl styrene/polypropylene glycol (MMA/BA/AMS/PPG) blocks. 4. The composition of claim 1, wherein the polymer matrix further comprises a random acrylic polymer. 5. The composition of claim 4, wherein the random acrylic polymer is made from one or more monomers selected from the group consisting of butyl acrylate, methyl acrylate, acrylic acid, ethyl hexyl acrylate, and hydroxy ethyl acrylate. 6. The composition of claim 5, wherein the random acrylic polymer is made from one or more monomers selected from the group consisting of ethyl hexyl acrylate, methyl acrylate, butyl acrylate, and octyl acrylamide. 7. The composition of claim 1, wherein the amphetamine is amphetamine free base. 8. The composition of claim 1, wherein the polymer matrix comprises from about 10% to about 20% by weight amphetamine free base. 9. The composition of claim 8, wherein the polymer matrix comprises from about 20% to about 80% by weight acrylic block copolymer. 10. The composition of claim 8, wherein the polymer matrix comprises from about 25% to about 65% by weight acrylic block copolymer. 11. The composition of claim 8, wherein the polymer matrix comprises from about 10% to about 55% by weight random acrylic polymer. 12. The composition of claim 8, wherein the polymer matrix comprises from about 20% to about 30% by weight random acrylic polymer. 13. The composition of claim 1, wherein the composition is capable of delivering amphetamine over a period of time of from about 6 to about 12 hours. 14. The composition of claim 1, wherein the composition delivers amphetamine over a period of time of up to about 10 hours. 15. The composition of claim 1, further comprising a backing layer. 16. The composition according to claim 15, further comprising a release liner. 17. A method for the transdermal delivery of amphetamine, comprising topically applying a composition as claimed in claim 1 to the skin or mucosa of a subject in need thereof. 18. A method of manufacturing a composition for the transdermal delivery of amphetamine in the form of a flexible finite system for topical application, comprising forming a polymer matrix blend comprising an acrylic block copolymer and amphetamine or pharmaceutically acceptable salt or prodrug thereof in a solvent, applying the polymer matrix blend to a support layer, and removing any remaining solvent. | Described are transdermal drug delivery compositions comprising amphetamine, methods of making them and therapeutic methods using them. The compositions are provided in a flexible, finite form (e.g. “patch”-type systems) and comprise a polymer matrix that includes amphetamine and an acrylic block copolymer.1. A composition for the transdermal delivery of amphetamine in the form of a flexible finite system for topical application, comprising a polymer matrix comprising amphetamine or a pharmaceutically acceptable salt or prodrug thereof, wherein the polymer matrix comprises an acrylic block copolymer. 2. The composition of claim 1, wherein the acrylic block copolymer is comprised of one or more monomers selected from the group consisting of methyl acrylate, methyl methacrylate, butyl acrylate, butyl methacrylate, hexyl acrylate, hexyl methacrylate, 2-ethylbutyl acrylate, 2-ethylbutyl methacrylate, isooctyl acrylate, isooctyl methacrylate, 2-ethylhexyl acrylate, 2-ethylhexyl methacrylate, decyl acrylate, decyl methacrylate, dodecyl acrylate, dodecyl methacrylate, tridecyl acrylate, tridecyl methacrylate, and octyl acrylamide. 2. The composition of claim 1, wherein the acrylic block copolymer comprises one or more of poly(butyl acrylate), poly(methyl methacrylate), methyl methacrylate/butyl acrylate (MMA/BA) blocks, and methyl methacrylate/butyl acrylate/alpha methyl styrene/polypropylene glycol (MMA/BA/AMS/PPG) blocks. 4. The composition of claim 1, wherein the polymer matrix further comprises a random acrylic polymer. 5. The composition of claim 4, wherein the random acrylic polymer is made from one or more monomers selected from the group consisting of butyl acrylate, methyl acrylate, acrylic acid, ethyl hexyl acrylate, and hydroxy ethyl acrylate. 6. The composition of claim 5, wherein the random acrylic polymer is made from one or more monomers selected from the group consisting of ethyl hexyl acrylate, methyl acrylate, butyl acrylate, and octyl acrylamide. 7. The composition of claim 1, wherein the amphetamine is amphetamine free base. 8. The composition of claim 1, wherein the polymer matrix comprises from about 10% to about 20% by weight amphetamine free base. 9. The composition of claim 8, wherein the polymer matrix comprises from about 20% to about 80% by weight acrylic block copolymer. 10. The composition of claim 8, wherein the polymer matrix comprises from about 25% to about 65% by weight acrylic block copolymer. 11. The composition of claim 8, wherein the polymer matrix comprises from about 10% to about 55% by weight random acrylic polymer. 12. The composition of claim 8, wherein the polymer matrix comprises from about 20% to about 30% by weight random acrylic polymer. 13. The composition of claim 1, wherein the composition is capable of delivering amphetamine over a period of time of from about 6 to about 12 hours. 14. The composition of claim 1, wherein the composition delivers amphetamine over a period of time of up to about 10 hours. 15. The composition of claim 1, further comprising a backing layer. 16. The composition according to claim 15, further comprising a release liner. 17. A method for the transdermal delivery of amphetamine, comprising topically applying a composition as claimed in claim 1 to the skin or mucosa of a subject in need thereof. 18. A method of manufacturing a composition for the transdermal delivery of amphetamine in the form of a flexible finite system for topical application, comprising forming a polymer matrix blend comprising an acrylic block copolymer and amphetamine or pharmaceutically acceptable salt or prodrug thereof in a solvent, applying the polymer matrix blend to a support layer, and removing any remaining solvent. | 1,600 |
1,011 | 15,427,952 | 1,618 | The present invention is directed to benzazepin-1-ol-derived compounds for use in the diagnosis of NMDA receptor-associated diseases or disorders by positron emission tomography (PET). The invention also relates to a method for the diagnosis of NMDA-receptor-associated diseases or disorders by administering to a patient in need of such diagnosis a radioactively labelled compound of the invention in an amount effective for PET imaging of NMDA receptors, recording at least one PET scan, and diagnosing an NMDA-receptor-associated disease or disorder from an abnormal NMDA receptor expression pattern on the PET scan. NMDA-receptor-associated diseases or disorders that can be diagnosed with the radioactively labelled benzazepin-1-ol-derived compounds include but are not limited to neurodegenerative diseases or disorders, Alzheimer's disease, depressive disorders, Parkinson's disease, traumatic brain injury, stroke, migraine, alcohol withdrawal and chronic and neuropathic pain. | 1. A method for the diagnosis of NMDA-receptor-associated diseases or disorders comprising the following steps:
(a) administering to a patient in need of such diagnosis a radioactively labelled compound in an amount effective for PET imaging of NMDA receptors, (b) recording at least one PET scan, and (c) diagnosing the patient as having an NMDA-receptor-associated disease or disorder from an abnormal NMDA receptor expression pattern on the PET scan, wherein the radioactively labelled compound has formula (I)
wherein
at least one atom of formula (I) is a radiolabeled atom, selected from the group of 11C-, 18F-, 13N-, or 15O-atom, suitable for positron emission tomography detection (PET);
one of R1, R2, R3 and R4 is independently selected from the group consisting of hydrogen, fluorine, —(C1-C4)alkyl, fluorinated —(C1-C4)alkyl, —CH2F,—CD2F, FCH2CH2—, FCH2CH2CH2—, —O(C1-C4)alkyl, fluorinated —O(C1-C4)alkyl, —OCH2F,—OCD2F, FCH2CH2O—, and FCH2CH2CH2O—,
and the other of R1 to R4 are hydrogen or fluorine;
R5 is selected from the group consisting of
hydrogen, —(C1-C6)alkyl, fluorinated —(C1-C6)alkyl, —CH2F,—CD2F, FCH2CH2—, and FCH2CH2CH2—;
Y is selected from the group consisting of
(C1-C6)alkyl, preferably C5-alkyl or C4-alkyl,
(C1-C6)alkoxyalkyl, (C1-C6)polyethyleneglycoyl, (CH2)2—O—(CH2)2—R6, —(CH2)3—O—R6, —(CH2)4—O—R6,
(C1-C6)heteroalkyl, —(CH2)3—X—R6 and —(CH2)4—X—R6, wherein X is sulfur or SO2,
—(CH2)3—CO—R6, —(CH2)2—CO—N(CH3)—CH2—R6, and —CO—(CH2)3—R6, wherein R8 is one or more hydrogen or fluorine;
R6 is selected from the group consisting of
substituted or non-substituted (C5-C6)aryl, substituted or non-substituted (C5-C6)heteroaryl, substituted or non-substituted phenyl or pyridyl, and
wherein Z is selected from the group consisting of hydrogen, fluorine and nitrile, wherein R7 is selected from the group consisting of hydrogen, fluorine, —(C1-C6)alkyl, fluorinated —(C1-C6)alkyl, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—;
and pharmaceutically acceptable salts or solvates thereof. 2. The method of diagnosis according to claim 1, wherein the at least one radiolabeled atom is a 11C-atom. 3. The method of diagnosis according to claim 1, wherein the NMDA-receptor-associated disease or disorder is selected from the group consisting of neurodegenerative diseases or disorders, Alzheimer's disease, depressive disorders, Parkinson's disease, traumatic brain injury, stroke, migraine, alcohol withdrawal and chronic and neuropathic pain. 4. The method of diagnosis according to claim 1, wherein at least one of R1, R3 or R5 comprise a 11C-atom. 5. The method of diagnosis according to claim 1, wherein one of R1, R3 is —O[11C]H3 or R5 is —[1C]H3. 6. The method of diagnosis according to claim 1, wherein one of R1, R2, R3 and R4 are independently selected from the group consisting of —CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2CH2—, —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O—, and the other of R1 to R4 are hydrogen or fluorine. 7. The method of diagnosis according to claim 1, wherein R1 is selected from the group consisting of —CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2CH2—, —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O and R2, R3 and R4 are hydrogen or fluorine 8. The method of diagnosis according to claim 1, wherein R3 is selected from the group consisting of —CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2CH2—, —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O and R1, R2 and R4 are hydrogen or fluorine. 9. The method of diagnosis according to claim 1, wherein R5 is selected from the group consisting of hydrogen, —CH2F,—CD2F, FCH2CH2—, and FCH2CH2CH2—. 10. The method of diagnosis according to claim 1, wherein Y is selected from the group consisting of —(CH2)i—R6 and —(CH2)e—O—(CH2)e—R6, wherein i is an integer from 2 to 6, and e is an integer from 1 to 3. 11. The method of diagnosis according to claim 1, wherein R6 is selected from the group consisting of phenyl,
wherein Z is hydrogen or nitrile and wherein R7 is selected from the group consisting of fluorine, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—. 12. The method of diagnosis according to claim 1, wherein the compound is R-configured at carbon 1. 13. The method of diagnosis according to claim 1, wherein the compound is S-configured at carbon 1. 14. The method of diagnosis according to claim 1, wherein
R1 is selected from the group consisting of —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O and R2, R3 and R4 are hydrogen, R5 is hydrogen, Y is —(CH2)4—R6, and R6 is substituted or un-substituted phenyl. 15. The method of diagnosis according to claim 1, wherein
R3 is selected from the group consisting of —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O and R1, R2 and R4 are hydrogen or fluorine, R5 is selected from the group consisting of hydrogen, —CH2F,—CD2F, FCH2CH2—, and FCH2CH2CH2—, Y is —(CH2)4—R6 or —(CH2)2—O—(CH2)2—R6, and R6 is selected from the group consisting of phenyl,
wherein Z is hydrogen or nitrile and wherein R7 is selected from the group consisting of fluorine, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—. 16. The method of diagnosis according to claim 1, wherein
R3 is selected from the group consisting of fluorine, —CH3, —CH2F,—CD2F, FCH2CH2— and FCH2CH2CH2— and R1, R2 and R4 are hydrogen, R5 is hydrogen, Y is —(CH2)4—R6, and R6 is substituted or un-substituted phenyl. 17. The method of diagnosis according to claim 1, wherein the compound is selected from the group consisting of
wherein
R9 is selected from the group consisting of CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—;
R10 is selected from the group consisting of hydrogen, CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—;
R11 is selected from the group consisting of fluorine, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—; and
Z is selected from the group consisting of hydrogen and nitrile. | The present invention is directed to benzazepin-1-ol-derived compounds for use in the diagnosis of NMDA receptor-associated diseases or disorders by positron emission tomography (PET). The invention also relates to a method for the diagnosis of NMDA-receptor-associated diseases or disorders by administering to a patient in need of such diagnosis a radioactively labelled compound of the invention in an amount effective for PET imaging of NMDA receptors, recording at least one PET scan, and diagnosing an NMDA-receptor-associated disease or disorder from an abnormal NMDA receptor expression pattern on the PET scan. NMDA-receptor-associated diseases or disorders that can be diagnosed with the radioactively labelled benzazepin-1-ol-derived compounds include but are not limited to neurodegenerative diseases or disorders, Alzheimer's disease, depressive disorders, Parkinson's disease, traumatic brain injury, stroke, migraine, alcohol withdrawal and chronic and neuropathic pain.1. A method for the diagnosis of NMDA-receptor-associated diseases or disorders comprising the following steps:
(a) administering to a patient in need of such diagnosis a radioactively labelled compound in an amount effective for PET imaging of NMDA receptors, (b) recording at least one PET scan, and (c) diagnosing the patient as having an NMDA-receptor-associated disease or disorder from an abnormal NMDA receptor expression pattern on the PET scan, wherein the radioactively labelled compound has formula (I)
wherein
at least one atom of formula (I) is a radiolabeled atom, selected from the group of 11C-, 18F-, 13N-, or 15O-atom, suitable for positron emission tomography detection (PET);
one of R1, R2, R3 and R4 is independently selected from the group consisting of hydrogen, fluorine, —(C1-C4)alkyl, fluorinated —(C1-C4)alkyl, —CH2F,—CD2F, FCH2CH2—, FCH2CH2CH2—, —O(C1-C4)alkyl, fluorinated —O(C1-C4)alkyl, —OCH2F,—OCD2F, FCH2CH2O—, and FCH2CH2CH2O—,
and the other of R1 to R4 are hydrogen or fluorine;
R5 is selected from the group consisting of
hydrogen, —(C1-C6)alkyl, fluorinated —(C1-C6)alkyl, —CH2F,—CD2F, FCH2CH2—, and FCH2CH2CH2—;
Y is selected from the group consisting of
(C1-C6)alkyl, preferably C5-alkyl or C4-alkyl,
(C1-C6)alkoxyalkyl, (C1-C6)polyethyleneglycoyl, (CH2)2—O—(CH2)2—R6, —(CH2)3—O—R6, —(CH2)4—O—R6,
(C1-C6)heteroalkyl, —(CH2)3—X—R6 and —(CH2)4—X—R6, wherein X is sulfur or SO2,
—(CH2)3—CO—R6, —(CH2)2—CO—N(CH3)—CH2—R6, and —CO—(CH2)3—R6, wherein R8 is one or more hydrogen or fluorine;
R6 is selected from the group consisting of
substituted or non-substituted (C5-C6)aryl, substituted or non-substituted (C5-C6)heteroaryl, substituted or non-substituted phenyl or pyridyl, and
wherein Z is selected from the group consisting of hydrogen, fluorine and nitrile, wherein R7 is selected from the group consisting of hydrogen, fluorine, —(C1-C6)alkyl, fluorinated —(C1-C6)alkyl, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—;
and pharmaceutically acceptable salts or solvates thereof. 2. The method of diagnosis according to claim 1, wherein the at least one radiolabeled atom is a 11C-atom. 3. The method of diagnosis according to claim 1, wherein the NMDA-receptor-associated disease or disorder is selected from the group consisting of neurodegenerative diseases or disorders, Alzheimer's disease, depressive disorders, Parkinson's disease, traumatic brain injury, stroke, migraine, alcohol withdrawal and chronic and neuropathic pain. 4. The method of diagnosis according to claim 1, wherein at least one of R1, R3 or R5 comprise a 11C-atom. 5. The method of diagnosis according to claim 1, wherein one of R1, R3 is —O[11C]H3 or R5 is —[1C]H3. 6. The method of diagnosis according to claim 1, wherein one of R1, R2, R3 and R4 are independently selected from the group consisting of —CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2CH2—, —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O—, and the other of R1 to R4 are hydrogen or fluorine. 7. The method of diagnosis according to claim 1, wherein R1 is selected from the group consisting of —CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2CH2—, —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O and R2, R3 and R4 are hydrogen or fluorine 8. The method of diagnosis according to claim 1, wherein R3 is selected from the group consisting of —CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2CH2—, —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O and R1, R2 and R4 are hydrogen or fluorine. 9. The method of diagnosis according to claim 1, wherein R5 is selected from the group consisting of hydrogen, —CH2F,—CD2F, FCH2CH2—, and FCH2CH2CH2—. 10. The method of diagnosis according to claim 1, wherein Y is selected from the group consisting of —(CH2)i—R6 and —(CH2)e—O—(CH2)e—R6, wherein i is an integer from 2 to 6, and e is an integer from 1 to 3. 11. The method of diagnosis according to claim 1, wherein R6 is selected from the group consisting of phenyl,
wherein Z is hydrogen or nitrile and wherein R7 is selected from the group consisting of fluorine, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—. 12. The method of diagnosis according to claim 1, wherein the compound is R-configured at carbon 1. 13. The method of diagnosis according to claim 1, wherein the compound is S-configured at carbon 1. 14. The method of diagnosis according to claim 1, wherein
R1 is selected from the group consisting of —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O and R2, R3 and R4 are hydrogen, R5 is hydrogen, Y is —(CH2)4—R6, and R6 is substituted or un-substituted phenyl. 15. The method of diagnosis according to claim 1, wherein
R3 is selected from the group consisting of —OCH3, —OCH2F,—OCD2F, FCH2CH2O— and FCH2CH2CH2O and R1, R2 and R4 are hydrogen or fluorine, R5 is selected from the group consisting of hydrogen, —CH2F,—CD2F, FCH2CH2—, and FCH2CH2CH2—, Y is —(CH2)4—R6 or —(CH2)2—O—(CH2)2—R6, and R6 is selected from the group consisting of phenyl,
wherein Z is hydrogen or nitrile and wherein R7 is selected from the group consisting of fluorine, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—. 16. The method of diagnosis according to claim 1, wherein
R3 is selected from the group consisting of fluorine, —CH3, —CH2F,—CD2F, FCH2CH2— and FCH2CH2CH2— and R1, R2 and R4 are hydrogen, R5 is hydrogen, Y is —(CH2)4—R6, and R6 is substituted or un-substituted phenyl. 17. The method of diagnosis according to claim 1, wherein the compound is selected from the group consisting of
wherein
R9 is selected from the group consisting of CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—;
R10 is selected from the group consisting of hydrogen, CH3, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—;
R11 is selected from the group consisting of fluorine, —CH2F,—CD2F, FCH2CH2—, FCH2CH2—, and FCH2CH2CH2—; and
Z is selected from the group consisting of hydrogen and nitrile. | 1,600 |
1,012 | 15,954,784 | 1,612 | Described herein, are compositions comprising an effective amount of a basic amino acid in free or salt form, and a water soluble strontium salt; and methods of making and using the same. | 1-11. (canceled) 12. A method to:
a. reduce or inhibit formation of dental caries, b. reduce, repair or inhibit early enamel lesions, c. reduce or inhibit demineralization and promote remineralization of the teeth, d. reduce hypersensitivity of the teeth, e. reduce or inhibit gingivitis, f. promote healing of sores or cuts in the mouth, g. reduce levels of acid producing bacteria, h. increase relative levels of arginolytic bacteria, i. inhibit microbial biofilm formation in the oral cavity, j. raise and/or maintain plaque pH at levels of at least pH 5.5 following sugar challenge, k. reduce plaque accumulation, l. treat dry mouth, m. whiten teeth, n. reduce erosion of the teeth, o. immunize the teeth against cariogenic bacteria and their effects, and/or p. clean the teeth and oral cavity; comprising applying an effective amount of an oral composition to a surface of the oral cavity of a subject in need thereof, the oral care composition comprising a. an effective amount of a basic amino acid, in free or salt form; and b. an effective amount of a water soluble strontium salt. 13. A method for promoting systemic health comprising applying an effective amount of an oral composition to a surface of the oral cavity of a subject in need thereof, the oral care composition comprising
a. an effective amount of a basic amino acid, in free or salt form; and b. an effective amount of a water soluble strontium salt. 14. (canceled) 15. A method according to claim 12, wherein the basic amino acid is arginine, or a salt thereof. 16. A method according to claim 12, wherein the water soluble strontium salt is selected from strontium acetate; strontium chloride; strontium nitrate; strontium lactate; strontium bromide; and a combination of two or more thereof. 17. A method according to claim 12, further comprising a fluoride ion source. 18. A method according to claim 17, wherein the fluoride ion source is selected from sodium monofluorophosphate; stannous fluoride; sodium fluoride; potassium fluoride; sodium fluorosilicate; ammonium fluorosilicate; amine fluoride; ammonium fluoride; and a combination of two or more thereof. 19. A method according to claim 12, wherein the composition is in the form of a dentifrice. 20. A method according to claim 12, wherein the composition is in the form of a mouth rinse. 21. A method according to claim 12, further comprising a potassium ion source. 22. A method according to claim 21, wherein said potassium ion source is selected from potassium nitrate and potassium chloride. 23. A method according to claim 12, further comprising a strontium-glass complex. 24. A method according to claim 23, wherein the strontium-glass complex comprises strontium sodium phosphosilicate. | Described herein, are compositions comprising an effective amount of a basic amino acid in free or salt form, and a water soluble strontium salt; and methods of making and using the same.1-11. (canceled) 12. A method to:
a. reduce or inhibit formation of dental caries, b. reduce, repair or inhibit early enamel lesions, c. reduce or inhibit demineralization and promote remineralization of the teeth, d. reduce hypersensitivity of the teeth, e. reduce or inhibit gingivitis, f. promote healing of sores or cuts in the mouth, g. reduce levels of acid producing bacteria, h. increase relative levels of arginolytic bacteria, i. inhibit microbial biofilm formation in the oral cavity, j. raise and/or maintain plaque pH at levels of at least pH 5.5 following sugar challenge, k. reduce plaque accumulation, l. treat dry mouth, m. whiten teeth, n. reduce erosion of the teeth, o. immunize the teeth against cariogenic bacteria and their effects, and/or p. clean the teeth and oral cavity; comprising applying an effective amount of an oral composition to a surface of the oral cavity of a subject in need thereof, the oral care composition comprising a. an effective amount of a basic amino acid, in free or salt form; and b. an effective amount of a water soluble strontium salt. 13. A method for promoting systemic health comprising applying an effective amount of an oral composition to a surface of the oral cavity of a subject in need thereof, the oral care composition comprising
a. an effective amount of a basic amino acid, in free or salt form; and b. an effective amount of a water soluble strontium salt. 14. (canceled) 15. A method according to claim 12, wherein the basic amino acid is arginine, or a salt thereof. 16. A method according to claim 12, wherein the water soluble strontium salt is selected from strontium acetate; strontium chloride; strontium nitrate; strontium lactate; strontium bromide; and a combination of two or more thereof. 17. A method according to claim 12, further comprising a fluoride ion source. 18. A method according to claim 17, wherein the fluoride ion source is selected from sodium monofluorophosphate; stannous fluoride; sodium fluoride; potassium fluoride; sodium fluorosilicate; ammonium fluorosilicate; amine fluoride; ammonium fluoride; and a combination of two or more thereof. 19. A method according to claim 12, wherein the composition is in the form of a dentifrice. 20. A method according to claim 12, wherein the composition is in the form of a mouth rinse. 21. A method according to claim 12, further comprising a potassium ion source. 22. A method according to claim 21, wherein said potassium ion source is selected from potassium nitrate and potassium chloride. 23. A method according to claim 12, further comprising a strontium-glass complex. 24. A method according to claim 23, wherein the strontium-glass complex comprises strontium sodium phosphosilicate. | 1,600 |
1,013 | 14,282,702 | 1,645 | This invention relates to a method of treating a dog for canine diseases comprising administering to the dog therapeutically effective amounts of a vaccine, wherein the vaccine comprises viral antigens, a bacterin, or both, and wherein the vaccine is administered subcutaneously or orally according to the schedules provided herein. | 1. A method of treating a dog for canine diseases comprising administering to the dog therapeutically effective amounts of vaccine, wherein the vaccine comprises viral antigens, a bacterin, or both, and wherein the vaccine is administered subcutaneously or orally in a first dose, orally in a second dose, orally in an optional third dose, and orally in one or more annual doses, and wherein the viral antigens comprise one or more of 1) canine distemper (CD) virus, 2) canine adenovirus type 2 (CAV-2), 3) canine parainfluenza (CPI) virus, 4) canine parvovirus (CPV), 5) and canine coronavirus (CCV), and wherein the bacterin comprises one or more bacteria selected from Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, L. pomona, L. bratislava, and Bordetella bronchiseptica; and any combination of viral antigens and bacteria thereof. 2. A method of treating a dog for canine diseases comprising administering to the dog therapeutically effective amounts of vaccine, wherein the vaccine comprises viral antigens, a bacterin, or both, and wherein the vaccine is administered subcutaneously in a first and in a second dose, and orally in a third dose, and orally in one or more annual doses, and wherein the viral antigens comprise one or more of 1) CD virus, 2) CAV-2, 3) CPI virus, 4) CPV, 5) and CCV, and the bacterin comprises one or more bacteria selected from Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, L. pomona, L. bratislava, and Bordetella bronchiseptica; and any combination of viral antigens and bacteria thereof. 3. A method according to claim 1 or 2, wherein the viral antigens are CD virus, CAV-2, CPI virus, and CPV. 4. A method according to claim 1 or 2, wherein the viral antigens are CD virus, CAV-2, CPI virus, and CPV, and the bacteria in the bacterin are Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, and L. pomona. 5. A method according to claim 1 or 2, wherein the viral antigens are CD virus, CAV-2, CPI virus, CPV, and CCV, and the bacteria in the bacterin are Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, and L. pomona. 6. A method according to claim 1 or 2, wherein the viral antigens are CD virus, CAV-2, CPI virus, and CPV, and the bacterium in the bacterin is Bordetella bronchiseptica. 7. A method according to claim 1 or 2, wherein the canine diseases comprise one or more of 1) CD caused by CD virus; 2) infectious canine hepatitis caused by CAV-1; 3) respiratory disease caused by CAV-2 or respiratory CCV; 4) CPI caused by CPI virus; 5) enteritis caused by CCV or CPV; 6) leptospirosis caused by Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, L. pomona, or L. Bratislava; and 7) infectious tracheobronchitis (“kennel cough”) caused by Bordetella bronchiseptica. 8. A method according to claim 7, wherein the diseases comprise 1) CD caused by CD virus; 2) infectious canine hepatitis caused by CAV-1; 3) respiratory disease caused by CAV-2; 4) CPI caused by CPI virus; 5) and canine parvoviral enteritis caused by CPV. 9. A method according to claim 1 or 2, wherein the viral antigens are present in the following ranges of amounts: for CD virus, about 102 TCID50 to about 108 TCID50, inclusive; for CAV-2, about 102 TCID50 to about 108 TCID50, inclusive; for CPV, about 103 TCID50 to about 1010 TCID50, inclusive; for CPI virus, about 103 TCID50 to about 1010 TCID50, inclusive; and for CCV, at least about 100 relative units (RU) per dose. 10. A method according to claim 1 or 2, wherein the viral antigens are present in the following ranges of amounts: for CD virus, about 103 TCID50 to about 106 TCID50, inclusive; for CAV-2, about 103 TCID50 to about 106 TCID50, inclusive; for CPV, about 106 TCID50 to about 109 TCID50, inclusive; for CPI virus, about 105 TCID50 to about 109 TCID50, inclusive; and for CCV, about 1,000 RU to about 4,500 RU per dose 11. A method according to claim 1 or 2, wherein the viral antigens are present in the following ranges of amounts: for CD virus, about 104 TCID50 to about 105 TCID50, inclusive; for CAV-2, about 104 TCID50 to about 105 TCID50, inclusive; for CPV, about 107 TCID50 to about 108 TCID50, inclusive; and for CPI virus, about 106 TCID50 to about 108 TCID50, inclusive. 12. A method according to claim 1 or 2, wherein each Leptospira is present in a range of amounts from about 100 nephelometric units (NU) to about 3,500 NU per vaccine dose, and wherein the Bordetella bronchiseptica is present in a range from about 3×106 to about 3×1011 cells inclusive. 13. A method according to claim 1 or 2, wherein each Leptospira is present in a range of amounts from about 200 NU to about 2,000 NU per dose, and wherein the Bordetella bronchiseptica is present in a range from about from about 3×107 to about 3×1010 cells inclusive. 14. A method according to claim 1 or 2, wherein the Bordetella bronchiseptica is present in a range from about 3×108 to about 3×109 cells inclusive. 15. A method according to claim 1 or 2, wherein the second dose is administered from 7 to 35 days, inclusive, after the first dose. 16. A method according to claim 15, wherein the second dose is administered about 3 weeks after the first dose. 17. A method according to claim 1 or 2, wherein the third dose is administered from 7 to 35 days, inclusive, after the second dose. 18. A method according to claim 17, wherein the third dose is administered about 3 weeks after the second dose. 19. A method according to claim 1 or 2, wherein the annual dose is administered about one year after the first dose. 20. A method according to claim 19, wherein annual doses administered after said annual dose are administered repeatedly about one year after the immediately prior annual dose. | This invention relates to a method of treating a dog for canine diseases comprising administering to the dog therapeutically effective amounts of a vaccine, wherein the vaccine comprises viral antigens, a bacterin, or both, and wherein the vaccine is administered subcutaneously or orally according to the schedules provided herein.1. A method of treating a dog for canine diseases comprising administering to the dog therapeutically effective amounts of vaccine, wherein the vaccine comprises viral antigens, a bacterin, or both, and wherein the vaccine is administered subcutaneously or orally in a first dose, orally in a second dose, orally in an optional third dose, and orally in one or more annual doses, and wherein the viral antigens comprise one or more of 1) canine distemper (CD) virus, 2) canine adenovirus type 2 (CAV-2), 3) canine parainfluenza (CPI) virus, 4) canine parvovirus (CPV), 5) and canine coronavirus (CCV), and wherein the bacterin comprises one or more bacteria selected from Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, L. pomona, L. bratislava, and Bordetella bronchiseptica; and any combination of viral antigens and bacteria thereof. 2. A method of treating a dog for canine diseases comprising administering to the dog therapeutically effective amounts of vaccine, wherein the vaccine comprises viral antigens, a bacterin, or both, and wherein the vaccine is administered subcutaneously in a first and in a second dose, and orally in a third dose, and orally in one or more annual doses, and wherein the viral antigens comprise one or more of 1) CD virus, 2) CAV-2, 3) CPI virus, 4) CPV, 5) and CCV, and the bacterin comprises one or more bacteria selected from Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, L. pomona, L. bratislava, and Bordetella bronchiseptica; and any combination of viral antigens and bacteria thereof. 3. A method according to claim 1 or 2, wherein the viral antigens are CD virus, CAV-2, CPI virus, and CPV. 4. A method according to claim 1 or 2, wherein the viral antigens are CD virus, CAV-2, CPI virus, and CPV, and the bacteria in the bacterin are Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, and L. pomona. 5. A method according to claim 1 or 2, wherein the viral antigens are CD virus, CAV-2, CPI virus, CPV, and CCV, and the bacteria in the bacterin are Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, and L. pomona. 6. A method according to claim 1 or 2, wherein the viral antigens are CD virus, CAV-2, CPI virus, and CPV, and the bacterium in the bacterin is Bordetella bronchiseptica. 7. A method according to claim 1 or 2, wherein the canine diseases comprise one or more of 1) CD caused by CD virus; 2) infectious canine hepatitis caused by CAV-1; 3) respiratory disease caused by CAV-2 or respiratory CCV; 4) CPI caused by CPI virus; 5) enteritis caused by CCV or CPV; 6) leptospirosis caused by Leptospira canicola, L. grippotyphosa, L. icterohaemorrhagiae, L. pomona, or L. Bratislava; and 7) infectious tracheobronchitis (“kennel cough”) caused by Bordetella bronchiseptica. 8. A method according to claim 7, wherein the diseases comprise 1) CD caused by CD virus; 2) infectious canine hepatitis caused by CAV-1; 3) respiratory disease caused by CAV-2; 4) CPI caused by CPI virus; 5) and canine parvoviral enteritis caused by CPV. 9. A method according to claim 1 or 2, wherein the viral antigens are present in the following ranges of amounts: for CD virus, about 102 TCID50 to about 108 TCID50, inclusive; for CAV-2, about 102 TCID50 to about 108 TCID50, inclusive; for CPV, about 103 TCID50 to about 1010 TCID50, inclusive; for CPI virus, about 103 TCID50 to about 1010 TCID50, inclusive; and for CCV, at least about 100 relative units (RU) per dose. 10. A method according to claim 1 or 2, wherein the viral antigens are present in the following ranges of amounts: for CD virus, about 103 TCID50 to about 106 TCID50, inclusive; for CAV-2, about 103 TCID50 to about 106 TCID50, inclusive; for CPV, about 106 TCID50 to about 109 TCID50, inclusive; for CPI virus, about 105 TCID50 to about 109 TCID50, inclusive; and for CCV, about 1,000 RU to about 4,500 RU per dose 11. A method according to claim 1 or 2, wherein the viral antigens are present in the following ranges of amounts: for CD virus, about 104 TCID50 to about 105 TCID50, inclusive; for CAV-2, about 104 TCID50 to about 105 TCID50, inclusive; for CPV, about 107 TCID50 to about 108 TCID50, inclusive; and for CPI virus, about 106 TCID50 to about 108 TCID50, inclusive. 12. A method according to claim 1 or 2, wherein each Leptospira is present in a range of amounts from about 100 nephelometric units (NU) to about 3,500 NU per vaccine dose, and wherein the Bordetella bronchiseptica is present in a range from about 3×106 to about 3×1011 cells inclusive. 13. A method according to claim 1 or 2, wherein each Leptospira is present in a range of amounts from about 200 NU to about 2,000 NU per dose, and wherein the Bordetella bronchiseptica is present in a range from about from about 3×107 to about 3×1010 cells inclusive. 14. A method according to claim 1 or 2, wherein the Bordetella bronchiseptica is present in a range from about 3×108 to about 3×109 cells inclusive. 15. A method according to claim 1 or 2, wherein the second dose is administered from 7 to 35 days, inclusive, after the first dose. 16. A method according to claim 15, wherein the second dose is administered about 3 weeks after the first dose. 17. A method according to claim 1 or 2, wherein the third dose is administered from 7 to 35 days, inclusive, after the second dose. 18. A method according to claim 17, wherein the third dose is administered about 3 weeks after the second dose. 19. A method according to claim 1 or 2, wherein the annual dose is administered about one year after the first dose. 20. A method according to claim 19, wherein annual doses administered after said annual dose are administered repeatedly about one year after the immediately prior annual dose. | 1,600 |
1,014 | 15,100,160 | 1,627 | Disclosed is sublingual administration of riluzole. In particular, a method for treating a neuropsychiatric disorder or symptom by administering a sublingual formulation of riluzole is provided. In addition, a method of relieving or reducing oral pain using the sublingual formulation of riluzole is disclosed. | 1. A sublingual formulation in the form of a lyophilized pharmaceutical composition that provides sublingual adsorption of riluzole comprising a pharmaceutically effective amount of riluzole or a pharmaceutically acceptable salt, solvate, anomer, enantiomer, hydrate or prodrug thereof, said formulation further comprising a tablet binder, a sugar alcohol selected from the group consisting of mannitol and sorbitol as a bulking agent, and docusate sodium as a solubilizing agent. 2. The sublingual formulation of claim 1 further comprising a flavoring agent. 3. The sublingual formulation of claim 1 further comprising a sweetener. 4. The sublingual formulation of claim 1 wherein the formulation riluzole comprises a riluzole prodrug. 5. The sublingual formulation of claim 3 said sweetener comprises sucralose. 6-30. (canceled) 31. A method of preparing a sublingual formulation of riluzole for treating a neuropsychiatric disorder or symptom in a human subject, said method comprising providing the composition of claim 1 in a pharmaceutically acceptable form. 32. The method of claim 31, wherein the neuropsychiatric disorder is anxiety disorders, mood disorders, neurodegenerative disorders, pain disorders, ALS, cognitive disorders, Huntington's disease, Parkinson's disease, supranuclear palsy, frontotemporal dementia, frontotemporal lobar degeneration, delirium, Alzheimer's disease, mild cognitive impairment, mild cognitive impairment due to Alzheimer's disease, ataxia, hereditary ataxia, depression, mania, attention deficit disorders, drug addiction, dementia, agitation, apathy, anxiety, psychoses, post-traumatic stress disorders, irritability, and disinhibition, learning disorders, memory loss, mental retardation, Rett Syndrome, tinnitus, personality disorders, bipolar disorders, obsessive-compulsive disorders, eating disorders, conduct disorders in DSM-5 and or combinations thereof. 33. The method of claim 31, wherein the neuropsychiatric symptom is anxiety, depression, stress, fatigue, feelings of panic, fear, uneasiness, problems in sleeping, cold or sweaty hands and/or feet, mood liability, mania, impaired concentration or attention, cognitive problems, obsessions, compulsions, repetitive behaviors, aggression, social phobias or impairments, stage fright, shortness of breath, heart palpitations, an inability to be still and calm, dry mouth, numbness or tingling in the hands or feet, nausea, muscle tension, dizziness apathy, elation, disinhibition, irritability, wandering, irritable bowel, belly pain, belly discomfort, diarrhea, change in bowel habits, abdominal bloating, abdominal, gas, abdominal bloating, constipation or combinations thereof. 34. The method of claim 31, wherein an effective dose of the sublingual formulation of riluzole is about 1 to 95% of that of the orally administered riluzole. 35. The method of claim 31, wherein the sublingual formulation of riluzole produces a rapid therapeutic onset of action within minutes. 36. The method of claim 31, wherein the sublingual formulation of riluzole is associated with minimal or no oral numbness, or said oral numbness dissipates in less than 30 minutes. 37. The method of claim 31, wherein the sublingual formulation delivers an exposure (AUC) similar to a higher orally administered dose of riluzole. 38. The method of claim 31, wherein the liver function abnormalities side effects from the sublingual formulation riluzole are attenuated relative to those which are associated with orally administered riluzole. 39. The method of claim 31, wherein the sublingual formulation of riluzole is dosed at or below about 50 mg/day, at or below about 35 mg/day, at or below about 17.5 mg/day, or at or below about 10 mg/day. 40. A method of preparing a sublingual formulation of riluzole for relieving or reducing oral pain in a human subject, said method comprising providing the composition of claim 1 in a pharmaceutically acceptable form for administration into the upper surface of the tongue, lips, buccal area, back of throat, entire oral cavity or combinations thereof. 41. The method of claim 40, wherein the sublingual formulation of riluzole is used to treat, oral pain caused by infection, inflammation, burn, cut, toothache, sore gums, canker sores, braces, minor dental procedures, denture irritation, oral surgery, neurologic disorders, disorders of the mucosa, oral ulcers, chemotherapy agents or combinations thereof. 42. The sublingual formulation of claim 1, wherein the sublingual formulation has a greater Cmax, an earlier or lesser Tmax, or a greater AUC per milligram of the riluzole than the same dose of orally administered riluzole. | Disclosed is sublingual administration of riluzole. In particular, a method for treating a neuropsychiatric disorder or symptom by administering a sublingual formulation of riluzole is provided. In addition, a method of relieving or reducing oral pain using the sublingual formulation of riluzole is disclosed.1. A sublingual formulation in the form of a lyophilized pharmaceutical composition that provides sublingual adsorption of riluzole comprising a pharmaceutically effective amount of riluzole or a pharmaceutically acceptable salt, solvate, anomer, enantiomer, hydrate or prodrug thereof, said formulation further comprising a tablet binder, a sugar alcohol selected from the group consisting of mannitol and sorbitol as a bulking agent, and docusate sodium as a solubilizing agent. 2. The sublingual formulation of claim 1 further comprising a flavoring agent. 3. The sublingual formulation of claim 1 further comprising a sweetener. 4. The sublingual formulation of claim 1 wherein the formulation riluzole comprises a riluzole prodrug. 5. The sublingual formulation of claim 3 said sweetener comprises sucralose. 6-30. (canceled) 31. A method of preparing a sublingual formulation of riluzole for treating a neuropsychiatric disorder or symptom in a human subject, said method comprising providing the composition of claim 1 in a pharmaceutically acceptable form. 32. The method of claim 31, wherein the neuropsychiatric disorder is anxiety disorders, mood disorders, neurodegenerative disorders, pain disorders, ALS, cognitive disorders, Huntington's disease, Parkinson's disease, supranuclear palsy, frontotemporal dementia, frontotemporal lobar degeneration, delirium, Alzheimer's disease, mild cognitive impairment, mild cognitive impairment due to Alzheimer's disease, ataxia, hereditary ataxia, depression, mania, attention deficit disorders, drug addiction, dementia, agitation, apathy, anxiety, psychoses, post-traumatic stress disorders, irritability, and disinhibition, learning disorders, memory loss, mental retardation, Rett Syndrome, tinnitus, personality disorders, bipolar disorders, obsessive-compulsive disorders, eating disorders, conduct disorders in DSM-5 and or combinations thereof. 33. The method of claim 31, wherein the neuropsychiatric symptom is anxiety, depression, stress, fatigue, feelings of panic, fear, uneasiness, problems in sleeping, cold or sweaty hands and/or feet, mood liability, mania, impaired concentration or attention, cognitive problems, obsessions, compulsions, repetitive behaviors, aggression, social phobias or impairments, stage fright, shortness of breath, heart palpitations, an inability to be still and calm, dry mouth, numbness or tingling in the hands or feet, nausea, muscle tension, dizziness apathy, elation, disinhibition, irritability, wandering, irritable bowel, belly pain, belly discomfort, diarrhea, change in bowel habits, abdominal bloating, abdominal, gas, abdominal bloating, constipation or combinations thereof. 34. The method of claim 31, wherein an effective dose of the sublingual formulation of riluzole is about 1 to 95% of that of the orally administered riluzole. 35. The method of claim 31, wherein the sublingual formulation of riluzole produces a rapid therapeutic onset of action within minutes. 36. The method of claim 31, wherein the sublingual formulation of riluzole is associated with minimal or no oral numbness, or said oral numbness dissipates in less than 30 minutes. 37. The method of claim 31, wherein the sublingual formulation delivers an exposure (AUC) similar to a higher orally administered dose of riluzole. 38. The method of claim 31, wherein the liver function abnormalities side effects from the sublingual formulation riluzole are attenuated relative to those which are associated with orally administered riluzole. 39. The method of claim 31, wherein the sublingual formulation of riluzole is dosed at or below about 50 mg/day, at or below about 35 mg/day, at or below about 17.5 mg/day, or at or below about 10 mg/day. 40. A method of preparing a sublingual formulation of riluzole for relieving or reducing oral pain in a human subject, said method comprising providing the composition of claim 1 in a pharmaceutically acceptable form for administration into the upper surface of the tongue, lips, buccal area, back of throat, entire oral cavity or combinations thereof. 41. The method of claim 40, wherein the sublingual formulation of riluzole is used to treat, oral pain caused by infection, inflammation, burn, cut, toothache, sore gums, canker sores, braces, minor dental procedures, denture irritation, oral surgery, neurologic disorders, disorders of the mucosa, oral ulcers, chemotherapy agents or combinations thereof. 42. The sublingual formulation of claim 1, wherein the sublingual formulation has a greater Cmax, an earlier or lesser Tmax, or a greater AUC per milligram of the riluzole than the same dose of orally administered riluzole. | 1,600 |
1,015 | 14,903,425 | 1,631 | A method of estimating transcript abundances includes: (a) obtaining transcript fragment sequencing data from a potential mixture of transcripts of a genetic locus of interest; (b) assigning this data to genetic coordinates of the locus of interest to obtain a data set of fragment genetic coordinate coverage and a coverage envelope curve; (c) setting a number of transcripts of the mixture; (d) pre-setting a probability distribution function of modelled genetic coverage for each transcript i composed of the product of a weight factor α i and the sum of at least 2 probability subfunctions j independently weighted by a weight factor β i,j ; (e) adding the probability distribution functions for each transcript to obtain a sum function; (f) fitting the sum function to the coverage envelope curve to optimize α i and β i,j to increase the fit; and (g) repeating steps (e) and (f) until a pre-set convergence criterion has been fulfilled. | 1. Method of estimating transcript abundances comprising the steps of:
a) obtaining transcript fragment sequencing data from a potential mixture of transcripts of a genetic locus of interest, b) assigning said fragment sequencing data to genetic coordinates of said locus of interest thereby obtaining a data set of fragment genetic coordinate coverage, said coverage for each genetic coordinate combined forming a coverage envelope curve, c) setting a number of transcripts of said mixture, d) pre-setting a probability distribution function of modelled genetic coverage for each transcript i, with i denoting the numerical identifier for a transcript, wherein said probability distribution function is defined by a weight factor al of said transcript i multiplied with the sum of at least 2 probability subfunctions j, with j denoting the numerical identifier for a probability subfunction, each probability subfunction j being independently weighted by a weight factor βi,j, e) adding the probability distribution functions of each transcript to obtain a sum function, f) fitting the sum function to the coverage envelope curve thereby optimizing the values for αi and βi,j to increase the fit, g) repeating steps e) and f) until a pre-set convergence criterion has been fulfilled, thereby obtaining the estimated transcript abundance for each transcript of the mixture given by the weight factor αi as optimized after the convergence criterion has been fulfilled. 2. The method of claim 1, wherein transcript fragment sequencing data comprises at least 5 transcript fragment sequences. 3. The method of claim 1, wherein the genetic locus of interest comprises one or more isoforms of one or more gene or genetic element, comprises at least two splice variants of one gene or genetic element. 4. The method of claim 1, wherein the step of setting a number of transcripts comprises obtaining pre-annotated sequence data from the genetic locus of interest and setting the number of transcripts to at least a number of different isoforms, including splice variants counting as different isoforms, expected from the genetic locus of interest. 5. The method of claim 1, wherein the probability subfunction j is constituted of positive values for each genetic coordinate. 6. The method of claim 1, wherein the probability subfunction j is selected from the group consisting of an aperiodic function, a Gaussian function, a square shape function, or a triangle shape function. 7. The method of claim 1, wherein the genetic coordinate corresponds to nucleotide positions in a genome, optionally transformed to omit genetic regions not of interest, wherein the genetic regions not of interest do not contain coverage by said transcript fragment sequencing data. 8. The method of claim 1, further comprising including a step b2) comprising removing genetic coordinate positions with splice junctions from said coverage envelope curve. 9. The method of claim 1, wherein said fragment genetic coordinate coverage contains the count of at least one nucleotide for each fragment sequence assigned to a genetic coordinate, wherein the at least one nucleotide comprises the fragment start site or the entire fragment sequence. 10. The method of claim 1, wherein the probability subfunctions for a transcript comprise maxima each at a different genetic coordinate. 11. The method of claim 1, wherein in step d) the probability subfunctions for a transcript are positioned or shifted in the genetic coordinate to cover the entire length of a transcript with a positive value. 12. The method of claim 1, comprising determining sequence reads of at least one transcript, wherein said reads comprise the sequence of fragments of said transcript, to provide said transcript fragment sequencing data. 13. The method of claim 1, wherein the transcript fragment sequences of said transcript fragment sequencing data have a length of 5 to 800 nucleotides. 14. The method of claim 1, wherein a full width at half maximum value for each probability subfunction for a transcript i is about identical. 15. A computer readable memory device comprising a computer program product for performing a method of claim 1 on a computer. 16. The method of claim 1, wherein the genetic locus of interest comprises at least two splice variants of one gene or genetic element. 17. The method of claim 1, wherein the probability subfunction j is a density function. 18. The method of claim 1, wherein the transcript fragment sequences of said transcript fragment sequencing data have a length of 12 to 70 nucleotides. 19. The method of claim 1, wherein the transcript fragment sequences of said transcript fragment sequencing data have a length of 9 to 150 nucleotides. 20. The method of claim 1, wherein the transcript fragment sequences of said transcript fragment sequencing data have a length of 7 to 400 nucleotides. | A method of estimating transcript abundances includes: (a) obtaining transcript fragment sequencing data from a potential mixture of transcripts of a genetic locus of interest; (b) assigning this data to genetic coordinates of the locus of interest to obtain a data set of fragment genetic coordinate coverage and a coverage envelope curve; (c) setting a number of transcripts of the mixture; (d) pre-setting a probability distribution function of modelled genetic coverage for each transcript i composed of the product of a weight factor α i and the sum of at least 2 probability subfunctions j independently weighted by a weight factor β i,j ; (e) adding the probability distribution functions for each transcript to obtain a sum function; (f) fitting the sum function to the coverage envelope curve to optimize α i and β i,j to increase the fit; and (g) repeating steps (e) and (f) until a pre-set convergence criterion has been fulfilled.1. Method of estimating transcript abundances comprising the steps of:
a) obtaining transcript fragment sequencing data from a potential mixture of transcripts of a genetic locus of interest, b) assigning said fragment sequencing data to genetic coordinates of said locus of interest thereby obtaining a data set of fragment genetic coordinate coverage, said coverage for each genetic coordinate combined forming a coverage envelope curve, c) setting a number of transcripts of said mixture, d) pre-setting a probability distribution function of modelled genetic coverage for each transcript i, with i denoting the numerical identifier for a transcript, wherein said probability distribution function is defined by a weight factor al of said transcript i multiplied with the sum of at least 2 probability subfunctions j, with j denoting the numerical identifier for a probability subfunction, each probability subfunction j being independently weighted by a weight factor βi,j, e) adding the probability distribution functions of each transcript to obtain a sum function, f) fitting the sum function to the coverage envelope curve thereby optimizing the values for αi and βi,j to increase the fit, g) repeating steps e) and f) until a pre-set convergence criterion has been fulfilled, thereby obtaining the estimated transcript abundance for each transcript of the mixture given by the weight factor αi as optimized after the convergence criterion has been fulfilled. 2. The method of claim 1, wherein transcript fragment sequencing data comprises at least 5 transcript fragment sequences. 3. The method of claim 1, wherein the genetic locus of interest comprises one or more isoforms of one or more gene or genetic element, comprises at least two splice variants of one gene or genetic element. 4. The method of claim 1, wherein the step of setting a number of transcripts comprises obtaining pre-annotated sequence data from the genetic locus of interest and setting the number of transcripts to at least a number of different isoforms, including splice variants counting as different isoforms, expected from the genetic locus of interest. 5. The method of claim 1, wherein the probability subfunction j is constituted of positive values for each genetic coordinate. 6. The method of claim 1, wherein the probability subfunction j is selected from the group consisting of an aperiodic function, a Gaussian function, a square shape function, or a triangle shape function. 7. The method of claim 1, wherein the genetic coordinate corresponds to nucleotide positions in a genome, optionally transformed to omit genetic regions not of interest, wherein the genetic regions not of interest do not contain coverage by said transcript fragment sequencing data. 8. The method of claim 1, further comprising including a step b2) comprising removing genetic coordinate positions with splice junctions from said coverage envelope curve. 9. The method of claim 1, wherein said fragment genetic coordinate coverage contains the count of at least one nucleotide for each fragment sequence assigned to a genetic coordinate, wherein the at least one nucleotide comprises the fragment start site or the entire fragment sequence. 10. The method of claim 1, wherein the probability subfunctions for a transcript comprise maxima each at a different genetic coordinate. 11. The method of claim 1, wherein in step d) the probability subfunctions for a transcript are positioned or shifted in the genetic coordinate to cover the entire length of a transcript with a positive value. 12. The method of claim 1, comprising determining sequence reads of at least one transcript, wherein said reads comprise the sequence of fragments of said transcript, to provide said transcript fragment sequencing data. 13. The method of claim 1, wherein the transcript fragment sequences of said transcript fragment sequencing data have a length of 5 to 800 nucleotides. 14. The method of claim 1, wherein a full width at half maximum value for each probability subfunction for a transcript i is about identical. 15. A computer readable memory device comprising a computer program product for performing a method of claim 1 on a computer. 16. The method of claim 1, wherein the genetic locus of interest comprises at least two splice variants of one gene or genetic element. 17. The method of claim 1, wherein the probability subfunction j is a density function. 18. The method of claim 1, wherein the transcript fragment sequences of said transcript fragment sequencing data have a length of 12 to 70 nucleotides. 19. The method of claim 1, wherein the transcript fragment sequences of said transcript fragment sequencing data have a length of 9 to 150 nucleotides. 20. The method of claim 1, wherein the transcript fragment sequences of said transcript fragment sequencing data have a length of 7 to 400 nucleotides. | 1,600 |
1,016 | 15,643,727 | 1,617 | Stable liquid formulations containing picosulfate and magnesium citrate are provided. The formulations are useful to treat constipation or for the clearance of the bowel prior to X-ray examination, endoscopy or surgery. | 1. A liquid formulation comprising:
picosulfate; magnesium citrate, and an antioxidant. 2. The liquid formulation of claim 1, wherein the picosulfate is sodium picosulfate. 3. The liquid formulation of claim 1, wherein the picosulfate is present in the formulation at a concentration in the range from about 0.10 mM to about 0.15 mM. 4. The liquid formulation claim 3, wherein the picosulfate is present in the formulation at a concentration in the range from about 0.12 mM to about 0.14 mM. 5. The liquid formulation of claim 1, wherein the antioxidant is chosen from ascorbic acid or a salt or ester thereof, butylated hydroxyanisole, butylated hydroxytoluene, histidine, D-mannose, propionic acid, potassium metabisulfite, propyl gallate, sodium metabisulfite, sodium sulfite, sodium thiosulfate, sulfur dioxide, and α-tocopherol. 6. The liquid formulation of claim 1, wherein the antioxidant is chosen from ascorbic acid, sodium ascorbate, sodium metabisulfite, potassium metabisulfite, sodium sulfite, sodium thiosulfate, and sulfur dioxide. 7. The liquid formulation of claim 1, wherein the antioxidant is sodium metabisulfite. 8. The liquid formulation of claim 1, wherein the antioxidant is chosen from butylated hydroxyanisole, butylated hydroxytoluene, propyl gallate, and α-tocopherol. 9. The liquid formulation of claim 1, wherein the antioxidant is present in the formulation at a concentration in the range from about 0.002 M to about 0.1 M. 10. The liquid formulation of claim 9, wherein the antioxidant is present in the formulation at a concentration in the range from about 0.005 M to about 0.02 M. 11. The liquid formulation of claim 1, wherein the magnesium citrate comprises magnesium and citrate in a molar ratio in the range from about 1:1 to about 1.5:1. 12. The liquid formulation of claim 11, wherein the magnesium citrate comprises magnesium and citrate in a molar ratio of about 1.4:1. 13. The liquid formulation of claim 1, wherein the magnesium citrate is formed from magnesium oxide and citric acid. 14. The liquid formulation of claim 1, wherein the magnesium citrate is present in the formulation at a concentration of magnesium in the range from about 0.2 M to about 0.8 M. 15. The liquid formulation of claim 14, wherein the magnesium citrate is present in the formulation at a concentration of magnesium in the range from about 0.5 M to about 0.6 M. 16. The liquid formulation of claim 1, further comprising a carboxylic acid. 17. The liquid formulation of claim 16, wherein the carboxylic acid is chosen from acetic acid, arginine, ascorbic acid, asparagine, aspartic acid, cysteine, fumaric acid, formic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, glycolic acid, histidine, lactic acid, lysine, maleic acid, malic acid, malonic acid, methionine, oxalic acid, propionic acid, serine, succinic acid, tartaric acid, threonine, and tyrosine. 18. The liquid formulation of claim 17, wherein the carboxylic acid is malic acid, malonic acid or gluconic acid. 19. The liquid formulation of claim 16, comprising the carboxylic acid at a concentration in the range from about 0.01 M to about 5 M. 20. The liquid formulation of claim 19, comprising the carboxylic acid at a concentration in the range from about 0.1 M to about 1 M. 21. The liquid formulation of claim 1, further comprising a water soluble anionic polymer. 22. The liquid formulation of claim 21, wherein the water soluble anionic polymer is chosen from alginic acid, carboxymethylcellulose, carrageenans, polyacrylic acid and copolymers thereof, and xanthan gum. 23. The liquid formulation of claim 21, comprising the water soluble anionic polymer at a concentration in the range from about 0.5 g/L to about 25 g/L. 24. The liquid formulation of claim 1, further comprising a preservative agent chosen from methyl paraben, propyl paraben, sodium benzoate and potassium benzoate. 25. The liquid formulation of claim 1, further comprising a chelating agent. 26. The liquid formulation of claim 1, wherein the pH of the formulation is in the range from about 4.5 to about 5.2. 27. The liquid formulation of claim 26, wherein the pH of the formulation is in the range from about 4.7 to about 4.9. 28. The liquid formulation of claim 27, wherein the pH of the formulation is about 4.9. 29. The liquid formulation of claim 1, wherein, when stored for 1 year at a temperature of about 25° C. and about 60% relative humidity, the formulation forms less than about 1.5% of Picosulfate Benzyl Alcohol and less than about 0.2% of Compound RRT 1.35 as measured by HPLC analysis area percentage compared to sodium picosulfate. 30. The liquid formulation of claim 1, wherein, when stored for 1 year at a temperature of about 25° C. and about 60% relative humidity, the formulation forms less than about 1.5% of Picosulfate Related Compound A, less than about 1.5% of Picosulfate Benzyl Alcohol, less than about 0.2% of Compound RRT 1.35, and less than about 0.2% of Compound RRT 1.94 as measured by HPLC analysis area percentage compared to sodium picosulfate. 31. The liquid formulation of claim 1, wherein, when stored for 1 year at a temperature of about 25° C. and about 60% relative humidity, the formulation forms less than about 1% of Picosulfate Related Compound A, less than about 1.3% of Picosulfate Benzyl Alcohol, less than about 0.2% of Compound RRT 1.35, and less than about 0.2% of Compound RRT 1.94 as measured by HPLC analysis area percentage compared to sodium picosulfate. | Stable liquid formulations containing picosulfate and magnesium citrate are provided. The formulations are useful to treat constipation or for the clearance of the bowel prior to X-ray examination, endoscopy or surgery.1. A liquid formulation comprising:
picosulfate; magnesium citrate, and an antioxidant. 2. The liquid formulation of claim 1, wherein the picosulfate is sodium picosulfate. 3. The liquid formulation of claim 1, wherein the picosulfate is present in the formulation at a concentration in the range from about 0.10 mM to about 0.15 mM. 4. The liquid formulation claim 3, wherein the picosulfate is present in the formulation at a concentration in the range from about 0.12 mM to about 0.14 mM. 5. The liquid formulation of claim 1, wherein the antioxidant is chosen from ascorbic acid or a salt or ester thereof, butylated hydroxyanisole, butylated hydroxytoluene, histidine, D-mannose, propionic acid, potassium metabisulfite, propyl gallate, sodium metabisulfite, sodium sulfite, sodium thiosulfate, sulfur dioxide, and α-tocopherol. 6. The liquid formulation of claim 1, wherein the antioxidant is chosen from ascorbic acid, sodium ascorbate, sodium metabisulfite, potassium metabisulfite, sodium sulfite, sodium thiosulfate, and sulfur dioxide. 7. The liquid formulation of claim 1, wherein the antioxidant is sodium metabisulfite. 8. The liquid formulation of claim 1, wherein the antioxidant is chosen from butylated hydroxyanisole, butylated hydroxytoluene, propyl gallate, and α-tocopherol. 9. The liquid formulation of claim 1, wherein the antioxidant is present in the formulation at a concentration in the range from about 0.002 M to about 0.1 M. 10. The liquid formulation of claim 9, wherein the antioxidant is present in the formulation at a concentration in the range from about 0.005 M to about 0.02 M. 11. The liquid formulation of claim 1, wherein the magnesium citrate comprises magnesium and citrate in a molar ratio in the range from about 1:1 to about 1.5:1. 12. The liquid formulation of claim 11, wherein the magnesium citrate comprises magnesium and citrate in a molar ratio of about 1.4:1. 13. The liquid formulation of claim 1, wherein the magnesium citrate is formed from magnesium oxide and citric acid. 14. The liquid formulation of claim 1, wherein the magnesium citrate is present in the formulation at a concentration of magnesium in the range from about 0.2 M to about 0.8 M. 15. The liquid formulation of claim 14, wherein the magnesium citrate is present in the formulation at a concentration of magnesium in the range from about 0.5 M to about 0.6 M. 16. The liquid formulation of claim 1, further comprising a carboxylic acid. 17. The liquid formulation of claim 16, wherein the carboxylic acid is chosen from acetic acid, arginine, ascorbic acid, asparagine, aspartic acid, cysteine, fumaric acid, formic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, glycolic acid, histidine, lactic acid, lysine, maleic acid, malic acid, malonic acid, methionine, oxalic acid, propionic acid, serine, succinic acid, tartaric acid, threonine, and tyrosine. 18. The liquid formulation of claim 17, wherein the carboxylic acid is malic acid, malonic acid or gluconic acid. 19. The liquid formulation of claim 16, comprising the carboxylic acid at a concentration in the range from about 0.01 M to about 5 M. 20. The liquid formulation of claim 19, comprising the carboxylic acid at a concentration in the range from about 0.1 M to about 1 M. 21. The liquid formulation of claim 1, further comprising a water soluble anionic polymer. 22. The liquid formulation of claim 21, wherein the water soluble anionic polymer is chosen from alginic acid, carboxymethylcellulose, carrageenans, polyacrylic acid and copolymers thereof, and xanthan gum. 23. The liquid formulation of claim 21, comprising the water soluble anionic polymer at a concentration in the range from about 0.5 g/L to about 25 g/L. 24. The liquid formulation of claim 1, further comprising a preservative agent chosen from methyl paraben, propyl paraben, sodium benzoate and potassium benzoate. 25. The liquid formulation of claim 1, further comprising a chelating agent. 26. The liquid formulation of claim 1, wherein the pH of the formulation is in the range from about 4.5 to about 5.2. 27. The liquid formulation of claim 26, wherein the pH of the formulation is in the range from about 4.7 to about 4.9. 28. The liquid formulation of claim 27, wherein the pH of the formulation is about 4.9. 29. The liquid formulation of claim 1, wherein, when stored for 1 year at a temperature of about 25° C. and about 60% relative humidity, the formulation forms less than about 1.5% of Picosulfate Benzyl Alcohol and less than about 0.2% of Compound RRT 1.35 as measured by HPLC analysis area percentage compared to sodium picosulfate. 30. The liquid formulation of claim 1, wherein, when stored for 1 year at a temperature of about 25° C. and about 60% relative humidity, the formulation forms less than about 1.5% of Picosulfate Related Compound A, less than about 1.5% of Picosulfate Benzyl Alcohol, less than about 0.2% of Compound RRT 1.35, and less than about 0.2% of Compound RRT 1.94 as measured by HPLC analysis area percentage compared to sodium picosulfate. 31. The liquid formulation of claim 1, wherein, when stored for 1 year at a temperature of about 25° C. and about 60% relative humidity, the formulation forms less than about 1% of Picosulfate Related Compound A, less than about 1.3% of Picosulfate Benzyl Alcohol, less than about 0.2% of Compound RRT 1.35, and less than about 0.2% of Compound RRT 1.94 as measured by HPLC analysis area percentage compared to sodium picosulfate. | 1,600 |
1,017 | 15,096,318 | 1,613 | A multi-part kit system comprising (i) a solid part A which comprises 10 to 80 wt. % of peroxy compound selected from the group consisting of KHSO 5 , K 2 S 2 O 8 , Na 2 S 2 O 8 , magnesium monoperoxyphthalate hexahydrate, sodium percarbonate and sodium perborate, 0.1 to 10 wt. % of LiCl, NaCl and/or KCl and 1 to 20 wt. % of H 2 N(CH 2 ) n SO 3 H with n=0, 1, 2 or 3, and (ii) a liquid part B in the form of an aqueous solution which comprises 0 to 20 wt. % of nonionic surfactant, 3.6 to 20 wt. % of amphoteric surfactant and 0.5 to 20 wt. % of at least one compound comprising substituted ammonium selected from the group consisting of dihydrocarbyl dimethylammonium chlorides or bromides, didecyl methyl-poly(oxyethyl) ammonium propionate, chlorhexidine gluconate, cetylpyridinium chloride or bromide, and polyhexamethylene biguanide hydrochloride, wherein at least one of the two hydrocarbyl residues comprises 8 to 18 carbon atoms. | 1. A multi-part kit system comprising:
(i) a solid part A which comprises 10 to 80 wt. % of at least one peroxy compound selected from the group consisting of KHSO5, K2S2O8, Na2S2O8, magnesium monoperoxyphthalate hexahydrate, sodium percarbonate, and sodium perborate; 0.1 to 10 wt. % of at least one MCI compound wherein M is selected from the group consisting of lithium, sodium and potassium; and 1 to 20 wt. % of at least one H2N(CH2)nSO3H compound with n=0, 1, 2 or 3, wherein the wt. % of part A ingredients is based on the total weight of solid part A; and, (ii) a liquid part B in the form of an aqueous solution which comprises 0 to 20 wt. % of nonionic surfactant; 3.6 to 20 wt. % of amphoteric surfactant; and 0.5 to 20 wt. % of at least one compound comprising substituted ammonium selected from the group consisting of dihydrocarbyl dimethylammonium halides, didecyl methyl-poly(oxyethyl) ammonium propionate, chlorhexidine gluconate, cetylpyridinium halide and polyhexamethylene biguanide hydrochloride, wherein at least one of the two hydrocarbyl residues comprises 8 to 18 carbon atoms, wherein halide means chloride or bromide, and wherein the wt. % of part B ingredients is based on the total weight of liquid part B. 2. The multi-part kit system of claim 1 consisting of said solid part A and said liquid part B. 3. The multi-part kit system of claim 1 or 2, wherein the solid part A is a flowable powder or takes the form of pellets or tablets. 4. The multi-part kit system of claim 1, wherein the solid part A comprises KHSO5. 5. The multi-part kit system of claim 4, wherein the solid part A comprises no other peroxy compound than KHSO5. 6. The multi-part kit system of claim 5, wherein the KHSO5 is present in the form of the triple salt 2KHSO5.KHSO4.K2SO4 and the proportion of said triple salt is 20 to 98.9 wt. %, based on the total weight of solid part A. 7. The multi-part kit system of claim 1, wherein solid part A comprises H2N(CH2)2SO3H. 8. The multi-part kit system of claim 1, wherein the solid part A comprises
36 to 87 wt. % of the triple salt 2KHSO5.KHSO4.K2SO4; 0.2 to 2.6 wt. % of NaCl; 3 to 20 wt. % of H2N(CH2)2SO3H; and 5 to 50 wt. % of one or more further additives selected from the group consisting of solid water-soluble inorganic fillers, solid pH-modifiers, dyes and peroxide decomposition stabilizers; wherein the sum of the wt. % totals 100 wt. %. 9. The multi-part kit system of claim 1, wherein the liquid part B comprises 25 to 95.9 wt. % of water. 10. The multi-part kit system of claim 1, wherein the liquid part B comprises one or more dihydrocarbyl dimethylammonium chlorides selected from the group consisting of didecyldimethylammonium chloride and benzalkonium chlorides of the formula C6H5CH2(CH3)2(CnH2n+1)N+Cl− with n=8, 10, 12, 14, 16 or 18. 11. The multi-part kit system of claim 10, wherein the liquid part B comprises none of the following compounds: didecyl methyl-poly(oxyethyl) ammonium propionate, chlorhexidine gluconate, cetylpyridinium chloride, cetylpyridinium bromide and polyhexamethylene biguanide hydrochloride. 12. The multi-part kit system of claim 1, wherein the liquid part B comprises
57 to 84 wt. % of water; 1 to 14 wt. % of nonionic surfactant; 6 to 20 wt. % of amphoteric surfactant; 2 to 18 wt. % of at least one compound selected from the group consisting of didecyldimethylammonium chloride and benzalkonium chlorides of the formula C6H5CH2(CH3)2(CnH2n+1)N+Cl− with n=8, 10, 12, 14, 16 or 18; and 1 to 6 wt. % of one or more further additives selected from the group consisting of MCI compounds; hard water sequestrants; corrosion inhibitors; water-soluble solvents and peroxide decomposition stabilizers; wherein the sum of the wt. % totals 100 wt. %. 13. The multi-part kit system of claim 1, wherein the solid part A comprises:
36 to 87 wt. % of the triple salt 2KHSO5.KHSO4.K2SO4; 0.2 to 2.6 wt. % of NaCl; 3 to 20 wt. % of H2N(CH2)2SO3H; and 5 to 50 wt. % of one or more further additives selected from the group consisting of solid water-soluble inorganic fillers, solid pH-modifiers, dyes and peroxide decomposition stabilizers; and wherein the liquid part B comprises: 57 to 84 wt. % of water; 1 to 14 wt. % of nonionic surfactant; 6 to 20 wt. % of amphoteric surfactant; 2 to 18 wt. % of at least one compound selected from the group consisting of didecyldimethylammonium chloride and benzalkonium chlorides of the formula C6H5CH2(CH3)2(CnH2n+1)N+Cl− with n=8, 10, 12, 14, 16 or 18; and 1 to 6 wt. % of one or more further additives selected from the group consisting of MCI compounds, hard water sequestrants, corrosion inhibitors, water-soluble solvents and peroxide decomposition stabilizers; wherein the sum of the wt. % of all ingredients in part A totals 100 wt. % and the sum of the wt. % of all ingredients in part B totals 100 wt. %. 14. The multi-part kit system of claim 1, wherein the parts A and B are such that upon mixing with each other (and water) the weight ratio between the at least one peroxy compound and the at least one compound comprising substituted ammonium is 4:1 to 23:1. 15. A process for the preparation of a RTU disinfectant comprising mixing all parts of the multi-part kit system of claim 1 and water in a ratio which ensures (i) a weight ratio between the at least one peroxy compound and the at least one compound comprising substituted ammonium in the range of 4:1 to 23:1; and (ii) a total content of 0.07 to 1.5 wt. % of the at least one peroxy compound plus the at least one compound comprising substituted ammonium based on the total weight of the RTU disinfectant. | A multi-part kit system comprising (i) a solid part A which comprises 10 to 80 wt. % of peroxy compound selected from the group consisting of KHSO 5 , K 2 S 2 O 8 , Na 2 S 2 O 8 , magnesium monoperoxyphthalate hexahydrate, sodium percarbonate and sodium perborate, 0.1 to 10 wt. % of LiCl, NaCl and/or KCl and 1 to 20 wt. % of H 2 N(CH 2 ) n SO 3 H with n=0, 1, 2 or 3, and (ii) a liquid part B in the form of an aqueous solution which comprises 0 to 20 wt. % of nonionic surfactant, 3.6 to 20 wt. % of amphoteric surfactant and 0.5 to 20 wt. % of at least one compound comprising substituted ammonium selected from the group consisting of dihydrocarbyl dimethylammonium chlorides or bromides, didecyl methyl-poly(oxyethyl) ammonium propionate, chlorhexidine gluconate, cetylpyridinium chloride or bromide, and polyhexamethylene biguanide hydrochloride, wherein at least one of the two hydrocarbyl residues comprises 8 to 18 carbon atoms.1. A multi-part kit system comprising:
(i) a solid part A which comprises 10 to 80 wt. % of at least one peroxy compound selected from the group consisting of KHSO5, K2S2O8, Na2S2O8, magnesium monoperoxyphthalate hexahydrate, sodium percarbonate, and sodium perborate; 0.1 to 10 wt. % of at least one MCI compound wherein M is selected from the group consisting of lithium, sodium and potassium; and 1 to 20 wt. % of at least one H2N(CH2)nSO3H compound with n=0, 1, 2 or 3, wherein the wt. % of part A ingredients is based on the total weight of solid part A; and, (ii) a liquid part B in the form of an aqueous solution which comprises 0 to 20 wt. % of nonionic surfactant; 3.6 to 20 wt. % of amphoteric surfactant; and 0.5 to 20 wt. % of at least one compound comprising substituted ammonium selected from the group consisting of dihydrocarbyl dimethylammonium halides, didecyl methyl-poly(oxyethyl) ammonium propionate, chlorhexidine gluconate, cetylpyridinium halide and polyhexamethylene biguanide hydrochloride, wherein at least one of the two hydrocarbyl residues comprises 8 to 18 carbon atoms, wherein halide means chloride or bromide, and wherein the wt. % of part B ingredients is based on the total weight of liquid part B. 2. The multi-part kit system of claim 1 consisting of said solid part A and said liquid part B. 3. The multi-part kit system of claim 1 or 2, wherein the solid part A is a flowable powder or takes the form of pellets or tablets. 4. The multi-part kit system of claim 1, wherein the solid part A comprises KHSO5. 5. The multi-part kit system of claim 4, wherein the solid part A comprises no other peroxy compound than KHSO5. 6. The multi-part kit system of claim 5, wherein the KHSO5 is present in the form of the triple salt 2KHSO5.KHSO4.K2SO4 and the proportion of said triple salt is 20 to 98.9 wt. %, based on the total weight of solid part A. 7. The multi-part kit system of claim 1, wherein solid part A comprises H2N(CH2)2SO3H. 8. The multi-part kit system of claim 1, wherein the solid part A comprises
36 to 87 wt. % of the triple salt 2KHSO5.KHSO4.K2SO4; 0.2 to 2.6 wt. % of NaCl; 3 to 20 wt. % of H2N(CH2)2SO3H; and 5 to 50 wt. % of one or more further additives selected from the group consisting of solid water-soluble inorganic fillers, solid pH-modifiers, dyes and peroxide decomposition stabilizers; wherein the sum of the wt. % totals 100 wt. %. 9. The multi-part kit system of claim 1, wherein the liquid part B comprises 25 to 95.9 wt. % of water. 10. The multi-part kit system of claim 1, wherein the liquid part B comprises one or more dihydrocarbyl dimethylammonium chlorides selected from the group consisting of didecyldimethylammonium chloride and benzalkonium chlorides of the formula C6H5CH2(CH3)2(CnH2n+1)N+Cl− with n=8, 10, 12, 14, 16 or 18. 11. The multi-part kit system of claim 10, wherein the liquid part B comprises none of the following compounds: didecyl methyl-poly(oxyethyl) ammonium propionate, chlorhexidine gluconate, cetylpyridinium chloride, cetylpyridinium bromide and polyhexamethylene biguanide hydrochloride. 12. The multi-part kit system of claim 1, wherein the liquid part B comprises
57 to 84 wt. % of water; 1 to 14 wt. % of nonionic surfactant; 6 to 20 wt. % of amphoteric surfactant; 2 to 18 wt. % of at least one compound selected from the group consisting of didecyldimethylammonium chloride and benzalkonium chlorides of the formula C6H5CH2(CH3)2(CnH2n+1)N+Cl− with n=8, 10, 12, 14, 16 or 18; and 1 to 6 wt. % of one or more further additives selected from the group consisting of MCI compounds; hard water sequestrants; corrosion inhibitors; water-soluble solvents and peroxide decomposition stabilizers; wherein the sum of the wt. % totals 100 wt. %. 13. The multi-part kit system of claim 1, wherein the solid part A comprises:
36 to 87 wt. % of the triple salt 2KHSO5.KHSO4.K2SO4; 0.2 to 2.6 wt. % of NaCl; 3 to 20 wt. % of H2N(CH2)2SO3H; and 5 to 50 wt. % of one or more further additives selected from the group consisting of solid water-soluble inorganic fillers, solid pH-modifiers, dyes and peroxide decomposition stabilizers; and wherein the liquid part B comprises: 57 to 84 wt. % of water; 1 to 14 wt. % of nonionic surfactant; 6 to 20 wt. % of amphoteric surfactant; 2 to 18 wt. % of at least one compound selected from the group consisting of didecyldimethylammonium chloride and benzalkonium chlorides of the formula C6H5CH2(CH3)2(CnH2n+1)N+Cl− with n=8, 10, 12, 14, 16 or 18; and 1 to 6 wt. % of one or more further additives selected from the group consisting of MCI compounds, hard water sequestrants, corrosion inhibitors, water-soluble solvents and peroxide decomposition stabilizers; wherein the sum of the wt. % of all ingredients in part A totals 100 wt. % and the sum of the wt. % of all ingredients in part B totals 100 wt. %. 14. The multi-part kit system of claim 1, wherein the parts A and B are such that upon mixing with each other (and water) the weight ratio between the at least one peroxy compound and the at least one compound comprising substituted ammonium is 4:1 to 23:1. 15. A process for the preparation of a RTU disinfectant comprising mixing all parts of the multi-part kit system of claim 1 and water in a ratio which ensures (i) a weight ratio between the at least one peroxy compound and the at least one compound comprising substituted ammonium in the range of 4:1 to 23:1; and (ii) a total content of 0.07 to 1.5 wt. % of the at least one peroxy compound plus the at least one compound comprising substituted ammonium based on the total weight of the RTU disinfectant. | 1,600 |
1,018 | 14,421,072 | 1,612 | Novel hydroxyalkyl methyl cellulose acetate succinates which a) have from 4.0 to less than 10.0 weight percent of succinoyl groups, b) have a weight average molecular weight M w of from 80,000 Dalton to 350,000 Dalton, and c) exhibit a turbidity of up to 41 NTU as a 1.5 weight percent solution in acetone; and novel hydroxyalkyl methyl cellulose acetate succinates which a) have from 10.0 to 20.0 weight percent of succinoyl groups, b) have a weight average molecular weight M w of from 80,000 Dalton to 350,000 Dalton, and c) exhibit a turbidity of up to 37 NTU as a 1.5 weight percent solution in acetone are useful for preparing solid dispersion of active ingredients in these hydroxyalkyl methyl cellulose acetate succinates. | 1. A hydroxyalkyl methyl cellulose acetate succinate
a) having from 4.0 to less than 10.0 weight percent of succinoyl groups, b) having a weight average molecular weight Mw of from 80,000 Dalton to 350,000 Dalton, measured by SEC-MALLS using as mobile phase a mixture of 40 parts by volume of acetonitrile and 60 parts by volume of aqueous buffer containing 50 mM NaH2PO4 and 0.1 M NaNO3, c) exhibiting a turbidity of up to 41 NTU as a 1.5 weight percent solution in acetone, measured according to USEPA method 180.1, and d) having a viscosity of up to 4.0 mPa·s, measured as a 2.0 wt % solution of the esterified cellulose ether in 0.43 wt.-% aqueous NaOH at 20° C. according to an Ubbelohde measurement according to DIN 51562-1:1999-01. 2. The hydroxyalkyl methyl cellulose acetate succinate of claim 1 exhibiting a turbidity of up to 37 NTU as a 1.5 weight percent solution in acetone. 3. The hydroxyalkyl methyl cellulose acetate succinate of claim 1 having e) from 10.0 to 14.0 weight percent of acetyl groups. 4. A hydroxyalkyl methyl cellulose acetate succinate
a) having from 10.0 to 20.0 weight percent of succinoyl groups, b) having a weight average molecular weight Mw of from 80,000 Dalton to 350,000 Dalton, measured by SEC-MALLS using as mobile phase a mixture of 40 parts by volume of acetonitrile and 60 parts by volume of aqueous buffer containing 50 mM NaH2PO4 and 0.1 M NaNO3, c) exhibiting a turbidity of up to 37 NTU as a 1.5 weight percent solution in acetone, measured according to USEPA method 180.1, and d) having a viscosity of up to 4.0 mPa·s, measured as a 2.0 wt % solution of the esterified cellulose ether in 0.43 wt.-% aqueous NaOH at 20° C. according to an Ubbelohde measurement according to DIN 51562-1:1999-01. 5. The hydroxyalkyl methyl cellulose acetate succinate of claim 4 having e) from 5.0 to 11.0 weight percent of acetyl groups. 6. The hydroxyalkyl methyl cellulose acetate succinate of claim 1 exhibiting a turbidity of from 10 to 30 NTU. 7. A composition comprising a liquid diluent and at least one hydroxyalkyl methyl cellulose acetate succinate of claim 1. 8. (canceled) 9. A solid dispersion comprising at least one active ingredient in at least one hydroxyalkyl methyl cellulose acetate succinate of claim 1. 10. The solid dispersion of claim 9 in the form of strands, pellets, granules, pills, tablets, caplets, microparticles, fillings of capsules or injection molded capsules or in the form of a powder, film, paste, cream, suspension or slurry. 11. A process for producing the solid dispersion of claim 9 comprising the steps of blending a) at least one hydroxyalkyl methyl cellulose acetate succinate of claim 1, b) one or more active ingredients and c) one or more optional additives, and subjecting the blend to extrusion. 12. A process for producing the solid dispersion of claim 9 comprising the steps of blending a) at least one hydroxyalkyl methyl cellulose acetate succinate of claim 1, b) one or more active ingredients, c) one or more optional additives, and d) a liquid diluent to prepare a liquid composition, and removing said liquid diluent. 13. The process of claim 12 wherein the liquid composition is subjected to spray-drying. 14. A dosage form being coated with the hydroxyalkyl methyl cellulose acetate succinate of claim 1. 15. A capsule shell comprising the hydroxyalkyl methyl cellulose acetate succinate of claim 1. 16. A composition comprising a liquid diluent and at least one hydroxyalkyl methyl cellulose acetate succinate of claim 4. 17. A solid dispersion comprising at least one active ingredient in at least one hydroxyalkyl methyl cellulose acetate succinate of claim 4. 18. A process for producing the solid dispersion of claim 17 comprising the steps of blending a) at least one hydroxyalkyl methyl cellulose acetate succinate of claim 4, b) one or more active ingredients and c) one or more optional additives, and subjecting the blend to extrusion. 19. process for producing the solid dispersion of claim 17 comprising the steps of blending a) at least one hydroxyalkyl methyl cellulose acetate succinate of claim 4, b) one or more active ingredients, c) one or more optional additives, and d) a liquid diluent to prepare a liquid composition, and removing said liquid diluent. 20. A dosage form being coated with the hydroxyalkyl methyl cellulose acetate succinate of claim 4. 21. A capsule shell comprising the hydroxyalkyl methyl cellulose acetate succinate of claim 4. | Novel hydroxyalkyl methyl cellulose acetate succinates which a) have from 4.0 to less than 10.0 weight percent of succinoyl groups, b) have a weight average molecular weight M w of from 80,000 Dalton to 350,000 Dalton, and c) exhibit a turbidity of up to 41 NTU as a 1.5 weight percent solution in acetone; and novel hydroxyalkyl methyl cellulose acetate succinates which a) have from 10.0 to 20.0 weight percent of succinoyl groups, b) have a weight average molecular weight M w of from 80,000 Dalton to 350,000 Dalton, and c) exhibit a turbidity of up to 37 NTU as a 1.5 weight percent solution in acetone are useful for preparing solid dispersion of active ingredients in these hydroxyalkyl methyl cellulose acetate succinates.1. A hydroxyalkyl methyl cellulose acetate succinate
a) having from 4.0 to less than 10.0 weight percent of succinoyl groups, b) having a weight average molecular weight Mw of from 80,000 Dalton to 350,000 Dalton, measured by SEC-MALLS using as mobile phase a mixture of 40 parts by volume of acetonitrile and 60 parts by volume of aqueous buffer containing 50 mM NaH2PO4 and 0.1 M NaNO3, c) exhibiting a turbidity of up to 41 NTU as a 1.5 weight percent solution in acetone, measured according to USEPA method 180.1, and d) having a viscosity of up to 4.0 mPa·s, measured as a 2.0 wt % solution of the esterified cellulose ether in 0.43 wt.-% aqueous NaOH at 20° C. according to an Ubbelohde measurement according to DIN 51562-1:1999-01. 2. The hydroxyalkyl methyl cellulose acetate succinate of claim 1 exhibiting a turbidity of up to 37 NTU as a 1.5 weight percent solution in acetone. 3. The hydroxyalkyl methyl cellulose acetate succinate of claim 1 having e) from 10.0 to 14.0 weight percent of acetyl groups. 4. A hydroxyalkyl methyl cellulose acetate succinate
a) having from 10.0 to 20.0 weight percent of succinoyl groups, b) having a weight average molecular weight Mw of from 80,000 Dalton to 350,000 Dalton, measured by SEC-MALLS using as mobile phase a mixture of 40 parts by volume of acetonitrile and 60 parts by volume of aqueous buffer containing 50 mM NaH2PO4 and 0.1 M NaNO3, c) exhibiting a turbidity of up to 37 NTU as a 1.5 weight percent solution in acetone, measured according to USEPA method 180.1, and d) having a viscosity of up to 4.0 mPa·s, measured as a 2.0 wt % solution of the esterified cellulose ether in 0.43 wt.-% aqueous NaOH at 20° C. according to an Ubbelohde measurement according to DIN 51562-1:1999-01. 5. The hydroxyalkyl methyl cellulose acetate succinate of claim 4 having e) from 5.0 to 11.0 weight percent of acetyl groups. 6. The hydroxyalkyl methyl cellulose acetate succinate of claim 1 exhibiting a turbidity of from 10 to 30 NTU. 7. A composition comprising a liquid diluent and at least one hydroxyalkyl methyl cellulose acetate succinate of claim 1. 8. (canceled) 9. A solid dispersion comprising at least one active ingredient in at least one hydroxyalkyl methyl cellulose acetate succinate of claim 1. 10. The solid dispersion of claim 9 in the form of strands, pellets, granules, pills, tablets, caplets, microparticles, fillings of capsules or injection molded capsules or in the form of a powder, film, paste, cream, suspension or slurry. 11. A process for producing the solid dispersion of claim 9 comprising the steps of blending a) at least one hydroxyalkyl methyl cellulose acetate succinate of claim 1, b) one or more active ingredients and c) one or more optional additives, and subjecting the blend to extrusion. 12. A process for producing the solid dispersion of claim 9 comprising the steps of blending a) at least one hydroxyalkyl methyl cellulose acetate succinate of claim 1, b) one or more active ingredients, c) one or more optional additives, and d) a liquid diluent to prepare a liquid composition, and removing said liquid diluent. 13. The process of claim 12 wherein the liquid composition is subjected to spray-drying. 14. A dosage form being coated with the hydroxyalkyl methyl cellulose acetate succinate of claim 1. 15. A capsule shell comprising the hydroxyalkyl methyl cellulose acetate succinate of claim 1. 16. A composition comprising a liquid diluent and at least one hydroxyalkyl methyl cellulose acetate succinate of claim 4. 17. A solid dispersion comprising at least one active ingredient in at least one hydroxyalkyl methyl cellulose acetate succinate of claim 4. 18. A process for producing the solid dispersion of claim 17 comprising the steps of blending a) at least one hydroxyalkyl methyl cellulose acetate succinate of claim 4, b) one or more active ingredients and c) one or more optional additives, and subjecting the blend to extrusion. 19. process for producing the solid dispersion of claim 17 comprising the steps of blending a) at least one hydroxyalkyl methyl cellulose acetate succinate of claim 4, b) one or more active ingredients, c) one or more optional additives, and d) a liquid diluent to prepare a liquid composition, and removing said liquid diluent. 20. A dosage form being coated with the hydroxyalkyl methyl cellulose acetate succinate of claim 4. 21. A capsule shell comprising the hydroxyalkyl methyl cellulose acetate succinate of claim 4. | 1,600 |
1,019 | 15,541,852 | 1,615 | This invention relates to the use of the combination of 25-hydroxyvitamin D3 (“25-OH D3”) and antioxidants/anti-inflammatories (ascorbic acid vitamin E and canthaxanthin) to make a premix or feed which can ameliorate various problems observed in poultry which have been subject to overfeeding. Feeds containing the 25-OH D3 and antioxidants/anti-inflammatories and premixes are also provided. | 1. A basal poultry feed additive composition comprising a combination of 25-Hydroxyvitamin D (25-OH D), Vitamin C, Vitamin E and canthaxathin. 2. A combination according to claim 1 further comprising at least one bio-active ingredient selected from the group consisting of Vitamin D, Vitamin B2, Vitamin B6, Niacin, Pantothenic Acid, Folic Acid, Biotin, Zinc, Copper, Manganese, and Selenium. 3. A combination according to claim 1 wherein the amount of Vitamin E or Vitamin C to Canthaxanthin may range from 40:1 to 1:1; preferably from 20:1 to 1:1; and more preferably from 10:1 to 1:1 4. A feed or premix comprising a combination according to claim 1. 5. A feed, or premix according to claim 1 which is suitable for poultry. 6. A combination, feed, or premix according to claim 1 wherein the 25-hydroxy vitamin D is 25-hydroxy vitamin D3 (25-OH D3). 7. A feed or according to claim 4 wherein:
the amount of 25-OH D3 is from 15-200μg/kg
the amount of Vitamin E is from 40-400 mg/kg
the amount of ascorbic acid is from 40-400 mg/kg; and
the amount of canthaxanthin is from 1-15 mg/kg 8. A feed according to claim 4 wherein:
the amount of 25-OH D3 is from 80-150 μg/kg;
the amount of Vitamin E is from 30-300 mg/kg;
the amount of ascorbic acid is from 80-300 mg/kg; and
the amount of canthaxanthin is from 1-15 mg/kg. 9. A feed according to claim 4 comprising:
25-OH D3: 35-150μg/kg;
Vitamin E: 80-300 mg/kg;
Canthaxanthin: 3-12 mg/kg; and
Ascorbic acid: 100-300 mg/kg. 10. A feed according to claim 4 comprising:
25-OH D3: 35 μg/kg
Vitamin E: 80 mg/kg
Canthaxanthin: 3 mg/kg; and
Ascorbic acid: 100 mg/kg. 11. A feed according to claim 4 comprising:
25-OH D3: 69 μg/kg
Vitamin E: 150 mg/kg
Canthaxanthin: 6 mg/kg; and
Ascorbic acid: 150 mg/kg. 12. A feed according to claim 4 comprising:
25-OH D3: 150μg/kg
Vitamin E: 300 mg/kg
Canthaxanthin: 12 mg/kg, and
Ascorbic acid: 300 mg/kg. 13. A premix which, when mixed with 1 kg of feed provides the feed of claim 4. | This invention relates to the use of the combination of 25-hydroxyvitamin D3 (“25-OH D3”) and antioxidants/anti-inflammatories (ascorbic acid vitamin E and canthaxanthin) to make a premix or feed which can ameliorate various problems observed in poultry which have been subject to overfeeding. Feeds containing the 25-OH D3 and antioxidants/anti-inflammatories and premixes are also provided.1. A basal poultry feed additive composition comprising a combination of 25-Hydroxyvitamin D (25-OH D), Vitamin C, Vitamin E and canthaxathin. 2. A combination according to claim 1 further comprising at least one bio-active ingredient selected from the group consisting of Vitamin D, Vitamin B2, Vitamin B6, Niacin, Pantothenic Acid, Folic Acid, Biotin, Zinc, Copper, Manganese, and Selenium. 3. A combination according to claim 1 wherein the amount of Vitamin E or Vitamin C to Canthaxanthin may range from 40:1 to 1:1; preferably from 20:1 to 1:1; and more preferably from 10:1 to 1:1 4. A feed or premix comprising a combination according to claim 1. 5. A feed, or premix according to claim 1 which is suitable for poultry. 6. A combination, feed, or premix according to claim 1 wherein the 25-hydroxy vitamin D is 25-hydroxy vitamin D3 (25-OH D3). 7. A feed or according to claim 4 wherein:
the amount of 25-OH D3 is from 15-200μg/kg
the amount of Vitamin E is from 40-400 mg/kg
the amount of ascorbic acid is from 40-400 mg/kg; and
the amount of canthaxanthin is from 1-15 mg/kg 8. A feed according to claim 4 wherein:
the amount of 25-OH D3 is from 80-150 μg/kg;
the amount of Vitamin E is from 30-300 mg/kg;
the amount of ascorbic acid is from 80-300 mg/kg; and
the amount of canthaxanthin is from 1-15 mg/kg. 9. A feed according to claim 4 comprising:
25-OH D3: 35-150μg/kg;
Vitamin E: 80-300 mg/kg;
Canthaxanthin: 3-12 mg/kg; and
Ascorbic acid: 100-300 mg/kg. 10. A feed according to claim 4 comprising:
25-OH D3: 35 μg/kg
Vitamin E: 80 mg/kg
Canthaxanthin: 3 mg/kg; and
Ascorbic acid: 100 mg/kg. 11. A feed according to claim 4 comprising:
25-OH D3: 69 μg/kg
Vitamin E: 150 mg/kg
Canthaxanthin: 6 mg/kg; and
Ascorbic acid: 150 mg/kg. 12. A feed according to claim 4 comprising:
25-OH D3: 150μg/kg
Vitamin E: 300 mg/kg
Canthaxanthin: 12 mg/kg, and
Ascorbic acid: 300 mg/kg. 13. A premix which, when mixed with 1 kg of feed provides the feed of claim 4. | 1,600 |
1,020 | 15,545,005 | 1,615 | The invention provides methods and compositions for the control and treatment of pests, insects, fungi, and nematodes in seeds, nuts, cereals, grains, plant growth media, plants, plant parts, and reproductive plant parts. One embodiment of the invention provides a composition comprising a seed, nut, cereal, or grain coated with a formulation. The formulation can comprise: (1) curcumin and a polar aprotic solvent; or (2) finely ground curcumin and a polar protic solvent. The formulation can further comprise carboxymethyl cellulose, one or more polymers or combinations thereof. The polar aprotic solvent can be dimethyl sulfoxide (DMSO) and the polar protic solvent can be water. | 1. (canceled) 2. The method of claim 8, wherein the curcumin solution further comprises carboxymethyl cellulose or a polymer. 3. The method of claim 8, wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO) and the polar protic solvent is water. 4. (canceled) 5. (canceled) 6. The method of claim 8, wherein the curcumin is present in the solution at about 0.1 mg/L−1 to about 400 mg/L−1. 7. (canceled) 8. A method of coating a cereal, seed, nut, or grain comprising:
(i) (a) combining curcumin with a polar aprotic solvent to form a curcumin solution; or
(b) combining finely ground curcumin with a polar protic solvent to form a curcumin solution; and
(ii) coating the cereal, seed, nut, or grain with the curcumin solution. 9. The method of claim 8, wherein the cereal, seed, nut or grain is further coated with one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof. 10. (canceled) 11. (canceled) 12. (canceled) 13. A method of cultivating a seed comprising:
(a) coating the seed with a formulation comprising (i) curcumin and a polar aprotic solvent; or (ii) finely ground curcumin and a polar protic solvent, and optionally, one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof; (b) contacting the seed with a medium suitable for germination; and (c) adding water. 14. (canceled) 15. A method for treating an infection or potential infection of a plant, part of a plant, reproductive plant material, or seed by fungi, insects or nematodes comprising contacting the plant, part of a plant, reproductive plant material, or seed with a formulation comprising (a) curcumin and a polar aprotic solvent; or (b) finely ground curcumin and a polar protic solvent, and optionally, one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof. 16. The method of claim 13, wherein the formulation further comprises carboxymethyl cellulose. 17. The method of claim 13, wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO) and the polar protic solvent is water. 18. The method of claim 9, wherein the one or more fungicides, one or more nematicides, one or more pesticides, or combinations thereof are not curcumin, and wherein the one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof are encapsulated in one or more liposomes. 19. The method of claim 8 further comprising coating the cereal, seed, nut, or grain with one or more liposomes containing one or more fungicides, nematicides or pesticides, one or more polymers, or combinations thereof. 20. The method of claim 13, wherein the curcumin is present in the formulation at about 0.1 mg/L−1 to about 400 mg/L−1. 21. The method of claim 13, wherein the seed is further coated with one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof. 22. The method of claim 21, wherein the one or more fungicides, one or more nematicides, one or more pesticides, or combinations thereof are not curcumin and wherein the with one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof are encapsulated in one or more liposomes. 23. The method of claim 13, further comprising coating the seed with one or more liposomes containing one or more fungicides, nematicides or pesticides, one or more polymers, or combinations thereof. 24. The method of claim 15, wherein the formulation further comprises carboxymethyl cellulose or a polymer. 25. The method of claim 15, wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO) and the polar protic solvent is water. 26. The method of claim 15, wherein the curcumin is present in the formulation at about 0.1 mg/L−1 to about 400 mg/L−1. 27. The method of claim 15, wherein the one or more fungicides, one or more nematicides, one or more pesticides, or combinations thereof are not curcumin, and wherein one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof are encapsulated in one or more liposomes. 28. The method of claim 15 further comprising contacting the plant, part of a plant, reproductive plant material, or seed with one or more liposomes containing one or more fungicides, nematicides or pesticides, one or more polymers, or combinations thereof. | The invention provides methods and compositions for the control and treatment of pests, insects, fungi, and nematodes in seeds, nuts, cereals, grains, plant growth media, plants, plant parts, and reproductive plant parts. One embodiment of the invention provides a composition comprising a seed, nut, cereal, or grain coated with a formulation. The formulation can comprise: (1) curcumin and a polar aprotic solvent; or (2) finely ground curcumin and a polar protic solvent. The formulation can further comprise carboxymethyl cellulose, one or more polymers or combinations thereof. The polar aprotic solvent can be dimethyl sulfoxide (DMSO) and the polar protic solvent can be water.1. (canceled) 2. The method of claim 8, wherein the curcumin solution further comprises carboxymethyl cellulose or a polymer. 3. The method of claim 8, wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO) and the polar protic solvent is water. 4. (canceled) 5. (canceled) 6. The method of claim 8, wherein the curcumin is present in the solution at about 0.1 mg/L−1 to about 400 mg/L−1. 7. (canceled) 8. A method of coating a cereal, seed, nut, or grain comprising:
(i) (a) combining curcumin with a polar aprotic solvent to form a curcumin solution; or
(b) combining finely ground curcumin with a polar protic solvent to form a curcumin solution; and
(ii) coating the cereal, seed, nut, or grain with the curcumin solution. 9. The method of claim 8, wherein the cereal, seed, nut or grain is further coated with one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof. 10. (canceled) 11. (canceled) 12. (canceled) 13. A method of cultivating a seed comprising:
(a) coating the seed with a formulation comprising (i) curcumin and a polar aprotic solvent; or (ii) finely ground curcumin and a polar protic solvent, and optionally, one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof; (b) contacting the seed with a medium suitable for germination; and (c) adding water. 14. (canceled) 15. A method for treating an infection or potential infection of a plant, part of a plant, reproductive plant material, or seed by fungi, insects or nematodes comprising contacting the plant, part of a plant, reproductive plant material, or seed with a formulation comprising (a) curcumin and a polar aprotic solvent; or (b) finely ground curcumin and a polar protic solvent, and optionally, one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof. 16. The method of claim 13, wherein the formulation further comprises carboxymethyl cellulose. 17. The method of claim 13, wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO) and the polar protic solvent is water. 18. The method of claim 9, wherein the one or more fungicides, one or more nematicides, one or more pesticides, or combinations thereof are not curcumin, and wherein the one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof are encapsulated in one or more liposomes. 19. The method of claim 8 further comprising coating the cereal, seed, nut, or grain with one or more liposomes containing one or more fungicides, nematicides or pesticides, one or more polymers, or combinations thereof. 20. The method of claim 13, wherein the curcumin is present in the formulation at about 0.1 mg/L−1 to about 400 mg/L−1. 21. The method of claim 13, wherein the seed is further coated with one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof. 22. The method of claim 21, wherein the one or more fungicides, one or more nematicides, one or more pesticides, or combinations thereof are not curcumin and wherein the with one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof are encapsulated in one or more liposomes. 23. The method of claim 13, further comprising coating the seed with one or more liposomes containing one or more fungicides, nematicides or pesticides, one or more polymers, or combinations thereof. 24. The method of claim 15, wherein the formulation further comprises carboxymethyl cellulose or a polymer. 25. The method of claim 15, wherein the polar aprotic solvent is dimethyl sulfoxide (DMSO) and the polar protic solvent is water. 26. The method of claim 15, wherein the curcumin is present in the formulation at about 0.1 mg/L−1 to about 400 mg/L−1. 27. The method of claim 15, wherein the one or more fungicides, one or more nematicides, one or more pesticides, or combinations thereof are not curcumin, and wherein one or more fungicides, one or more nematicides, one or more pesticides, one or more polymers, or combinations thereof are encapsulated in one or more liposomes. 28. The method of claim 15 further comprising contacting the plant, part of a plant, reproductive plant material, or seed with one or more liposomes containing one or more fungicides, nematicides or pesticides, one or more polymers, or combinations thereof. | 1,600 |
1,021 | 15,079,735 | 1,653 | Provided is a method of culturing one or more microorganisms. The method includes culturing one or more microorganisms in a medium comprising crude glycerol at a first concentration level, feeding to the media an additional amount of crude glycerol, once the first concentration of glycerol is reduced to a first threshold level, at a concentration sufficient to achieve the first concentration level, monitoring the crude glycerol concentration until the first concentration level of the crude glycerol is reduced to the first threshold level. The steps may be repeated until a desired microorganism cell density is achieved. | 1. A method of culturing one or more microorganisms, comprising:
(a) culturing one or more microorganisms in a medium comprising crude glycerol at a first concentration level; (b) feeding to the media an additional amount of crude glycerol, once the first concentration of glycerol is reduced to a first threshold level, at a concentration sufficient to achieve the first concentration level; (c) monitoring the crude glycerol concentration until the first concentration level of the crude glycerol is reduced to the first threshold level; and (d) repeating steps (b) (c) and (d) until a desired microorganism cell density is achieved. 2. The method of claim 1, wherein the one or more microorganisms is of the genus Thraustochytrium or the genus Schizochytrium. 3. The method of claim 1, wherein the one or more microorganisms is ONC-T18. 4. The method of claim 1, wherein the microorganisms are capable of producing one or more fatty acids. 5. The method of claim 4, wherein the fatty acids are polyunsaturated fatty acids. 6. The method of claim 5, wherein the polyunsaturated fatty acids are selected from the group consisting of alpha linolenic acid, arachidonic acid, docosahexanenoic acid, docosapentaenoic acid, eicosapentaenoic acid, gamma-linolenic acid, linoleic acid, linolenic acid, and combinations thereof. 7. The method of claim 4, wherein the method further comprises isolating the fatty acids. 8. The method of claim 1, wherein the first concentration level is between 1 and 60 g/L. 9. The method of claim 1, wherein the first threshold level is between 0 and 5 g/L. 10. The method of claim 1, wherein the monitoring comprises measuring dissolved oxygen levels. 11. The method of claim 1, wherein the monitoring comprises obtaining a sample of the media and determining the glycerol concentration in the sample. 12. The method of claim 11, wherein the monitoring comprises analyzing the sample using calorimetric assay, chemical reaction based calorimetric assay, fluorescence assay, HPLC assay, enzymatic assay, or combinations thereof. 13. The method of claim 1, wherein the cell density is between 50 g/L and 250 g/L. 14. The method of claim 1, wherein the microorganism cell density contains 50% to 80% by weight of total fatty acids. 15. The method of claim 1, wherein the total fatty acids comprise 10 to 45% DHA. 16. The method of claim 1, wherein the microorganism cell density contains 5 to 36% DHA by total cell weight. 17. The method of claim 1, wherein the medium comprises one or more additional carbon sources. 18. The method of claim 1, wherein the crude glycerol is not sterilized. 19. The method of claim 1, wherein the crude glycerol comprises methanol, water, ash, non-glycerin organic matter, sodium sulphate, methyl tallowate, or a combination thereof. 20. The method of claim 19, wherein the ash comprises calcium, iron, magnesium, potassium, sodium, zinc or a combination thereof. 21. The method of claim 1, wherein the crude glycerol is a biodiesel byproduct. 22. A method for producing one or more fatty acids, comprising:
(a) providing a microorganism capable of producing fatty acids; (b) providing a media comprising crude glycerol; and (c) culturing the microorganism in the media under sufficient conditions to produce the one or more fatty acids to provide a final concentration of the one or more unsaturated fatty acids that is at least 50% by weight of the microorganisms. 23. The method of claim 22, wherein the microorganism is of the genus Thraustochytrium or the genus Schizochytrium. 24. The method of claim 22, wherein the microorganism is ONC-T18. 25. The method of claim 22, wherein the fatty acids comprise polyunsaturated fatty acids. 26. The method of claim 25, wherein the polyunsaturated fatty acids are selected from the group consisting of alpha linolenic acid, arachidonic acid, docosahexanenoic acid, docosapentaenoic acid, eicosapentaenoic acid, gamma-linolenic acid, linoleic acid, linolenic acid, and combinations thereof. 27. The method of claim 22, further comprising isolating the fatty acids. 28. The method of claim 22, wherein the media comprises one or more additional carbon sources. 29. The method of claim 22, wherein the crude glycerol is not sterilized. 30. The method of claim 22, wherein the crude glycerol comprises methanol, water, ash, non-glycerin organic matter, sodium sulphate, methyl tallowate, or a combination thereof. 31. The method of claim 30, wherein the ash comprises calcium, iron, magnesium, potassium, sodium, zinc or a combination thereof. 32. The method of claim 22, wherein the crude glycerol is a biodiesel byproduct. | Provided is a method of culturing one or more microorganisms. The method includes culturing one or more microorganisms in a medium comprising crude glycerol at a first concentration level, feeding to the media an additional amount of crude glycerol, once the first concentration of glycerol is reduced to a first threshold level, at a concentration sufficient to achieve the first concentration level, monitoring the crude glycerol concentration until the first concentration level of the crude glycerol is reduced to the first threshold level. The steps may be repeated until a desired microorganism cell density is achieved.1. A method of culturing one or more microorganisms, comprising:
(a) culturing one or more microorganisms in a medium comprising crude glycerol at a first concentration level; (b) feeding to the media an additional amount of crude glycerol, once the first concentration of glycerol is reduced to a first threshold level, at a concentration sufficient to achieve the first concentration level; (c) monitoring the crude glycerol concentration until the first concentration level of the crude glycerol is reduced to the first threshold level; and (d) repeating steps (b) (c) and (d) until a desired microorganism cell density is achieved. 2. The method of claim 1, wherein the one or more microorganisms is of the genus Thraustochytrium or the genus Schizochytrium. 3. The method of claim 1, wherein the one or more microorganisms is ONC-T18. 4. The method of claim 1, wherein the microorganisms are capable of producing one or more fatty acids. 5. The method of claim 4, wherein the fatty acids are polyunsaturated fatty acids. 6. The method of claim 5, wherein the polyunsaturated fatty acids are selected from the group consisting of alpha linolenic acid, arachidonic acid, docosahexanenoic acid, docosapentaenoic acid, eicosapentaenoic acid, gamma-linolenic acid, linoleic acid, linolenic acid, and combinations thereof. 7. The method of claim 4, wherein the method further comprises isolating the fatty acids. 8. The method of claim 1, wherein the first concentration level is between 1 and 60 g/L. 9. The method of claim 1, wherein the first threshold level is between 0 and 5 g/L. 10. The method of claim 1, wherein the monitoring comprises measuring dissolved oxygen levels. 11. The method of claim 1, wherein the monitoring comprises obtaining a sample of the media and determining the glycerol concentration in the sample. 12. The method of claim 11, wherein the monitoring comprises analyzing the sample using calorimetric assay, chemical reaction based calorimetric assay, fluorescence assay, HPLC assay, enzymatic assay, or combinations thereof. 13. The method of claim 1, wherein the cell density is between 50 g/L and 250 g/L. 14. The method of claim 1, wherein the microorganism cell density contains 50% to 80% by weight of total fatty acids. 15. The method of claim 1, wherein the total fatty acids comprise 10 to 45% DHA. 16. The method of claim 1, wherein the microorganism cell density contains 5 to 36% DHA by total cell weight. 17. The method of claim 1, wherein the medium comprises one or more additional carbon sources. 18. The method of claim 1, wherein the crude glycerol is not sterilized. 19. The method of claim 1, wherein the crude glycerol comprises methanol, water, ash, non-glycerin organic matter, sodium sulphate, methyl tallowate, or a combination thereof. 20. The method of claim 19, wherein the ash comprises calcium, iron, magnesium, potassium, sodium, zinc or a combination thereof. 21. The method of claim 1, wherein the crude glycerol is a biodiesel byproduct. 22. A method for producing one or more fatty acids, comprising:
(a) providing a microorganism capable of producing fatty acids; (b) providing a media comprising crude glycerol; and (c) culturing the microorganism in the media under sufficient conditions to produce the one or more fatty acids to provide a final concentration of the one or more unsaturated fatty acids that is at least 50% by weight of the microorganisms. 23. The method of claim 22, wherein the microorganism is of the genus Thraustochytrium or the genus Schizochytrium. 24. The method of claim 22, wherein the microorganism is ONC-T18. 25. The method of claim 22, wherein the fatty acids comprise polyunsaturated fatty acids. 26. The method of claim 25, wherein the polyunsaturated fatty acids are selected from the group consisting of alpha linolenic acid, arachidonic acid, docosahexanenoic acid, docosapentaenoic acid, eicosapentaenoic acid, gamma-linolenic acid, linoleic acid, linolenic acid, and combinations thereof. 27. The method of claim 22, further comprising isolating the fatty acids. 28. The method of claim 22, wherein the media comprises one or more additional carbon sources. 29. The method of claim 22, wherein the crude glycerol is not sterilized. 30. The method of claim 22, wherein the crude glycerol comprises methanol, water, ash, non-glycerin organic matter, sodium sulphate, methyl tallowate, or a combination thereof. 31. The method of claim 30, wherein the ash comprises calcium, iron, magnesium, potassium, sodium, zinc or a combination thereof. 32. The method of claim 22, wherein the crude glycerol is a biodiesel byproduct. | 1,600 |
1,022 | 15,521,970 | 1,633 | Disclosed are methods for coating microcarriers with a marrow stromal cell derived extracellular matrix, and maintaining and expanding mammalian mesenchymal stem cells on the marrow stromal cell derived extracellular matrix coated microcarriers in culture. | 1. A method of maintaining and expanding mammalian mesenchymal stem cells in culture in an undifferentiated state, the method comprising:
a. producing a 3D extracellular matrix coating on the surface of microcarriers comprising:
i. adding the microcarriers to a culture medium;
ii. adding mammalian bone marrow stromal cells to the culture medium;
iii. culturing the bone marrow stromal cells to produce the 3D extracellular matrix coating on the surface of the microcarriers;
iv. decellularizing the extracellular matrix coated microcarriers of the bone marrow stromal cells; and
b. culturing the mammalian mesenchymal stem cells in the presence of the extracellular matrix coated microcarriers; wherein the extracellular matrix coating restrains differentiation of the mammalian mesenchymal stem cells. 2. The method of claim 1, wherein the extracellular matrix coating comprises collagen alpha-1 (XII), collagen alpha-3 (VI), EMILIN-1, serpin H1, thrombospondin-1, tenascin precursor (TN) (Human), transforming growth factor-beta-induced protein, and vimentin. 3. The method of claim 2, wherein the extracellular matrix coating further comprises type I collagen, type III collagen, fibronectin, decorin, biglycan, perlecan, and laminin. 4. The method of claim 3, wherein the extracellular matrix coating further comprises at least one of syndecan-1, collagen type V, or collagen type VI. 5-6. (canceled) 7. The method of claim 1, wherein the bone marrow stromal cells are isolated bone marrow mesenchymal stem cells. 8. The method of claim 1, wherein the mammalian mesenchymal stem cells are obtained from bone marrow or umbilical cord blood. 9. (canceled) 10. The method of the method of claim 1, wherein the microcarriers are spherical in shape. 11. The method of the method of claim 1, wherein the microcarriers have a positive charge. 12. (canceled) 13. The method of claim 11, wherein the microcarriers comprise are comprised of a cross-linked dextran matrix. 14. The method of the method of claim 1, wherein the microcarriers are cylindrical in shape. 15. The method of claim 14, wherein the microcarriers are hollow fibers. 16. (canceled) 17. The method of the method of claim 1, wherein the method further comprises culturing the bone marrow stromal cells under normoxic conditions. 18. The method of the method of claim 1, wherein the method further comprises culturing the mammalian mesenchymal stem cells under normoxic conditions. 19-22. (canceled) 23. A method of maintaining and expanding mammalian mesenchymal stem cells in culture in an undifferentiated state, the method comprising:
a. obtaining bone marrow stromal cell derived 3D extracellular matrix coated microcarriers; and b. culturing the mammalian mesenchymal stem cells in the presence of the extracellular matrix coated microcarriers, wherein the extracellular matrix coating restrains differentiation of the mammalian mesenchymal stem cells. 24. The method of claim 23, wherein the extracellular matrix coating comprises collagen alpha-1(XII), collagen alpha-3(VI), EMILIN-1, serpin H1, thrombospondin-1, tenascin precursor (TN) (Human), transforming growth factor-beta-induced protein, and vimentin. 25. The method of claim 24, wherein the extracellular matrix coating further comprises type I collagen, type III collagen, fibronectin, decorin, biglycan, perlecan, and laminin. 26. The method of claim 25, wherein the extracellular matrix coating further comprises at least one of syndecan-1, collagen type V, or collagen type VI. 27-44. (canceled) 45. A plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers. 46. The plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers of claim 45, further comprising mammalian mesenchymal stem cells attached to the plurality of extracellular matrix coated microcarriers. 47. (canceled) 48. The plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers of claim 45, wherein the extracellular matrix coated microcarriers are free or are substantially free of bone marrow stromal cells. 49. The plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers of claim 45, wherein the extracellular matrix coated microcarriers are comprised in a composition. 50. The plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers of claim 49, wherein the composition is a cell culture media. | Disclosed are methods for coating microcarriers with a marrow stromal cell derived extracellular matrix, and maintaining and expanding mammalian mesenchymal stem cells on the marrow stromal cell derived extracellular matrix coated microcarriers in culture.1. A method of maintaining and expanding mammalian mesenchymal stem cells in culture in an undifferentiated state, the method comprising:
a. producing a 3D extracellular matrix coating on the surface of microcarriers comprising:
i. adding the microcarriers to a culture medium;
ii. adding mammalian bone marrow stromal cells to the culture medium;
iii. culturing the bone marrow stromal cells to produce the 3D extracellular matrix coating on the surface of the microcarriers;
iv. decellularizing the extracellular matrix coated microcarriers of the bone marrow stromal cells; and
b. culturing the mammalian mesenchymal stem cells in the presence of the extracellular matrix coated microcarriers; wherein the extracellular matrix coating restrains differentiation of the mammalian mesenchymal stem cells. 2. The method of claim 1, wherein the extracellular matrix coating comprises collagen alpha-1 (XII), collagen alpha-3 (VI), EMILIN-1, serpin H1, thrombospondin-1, tenascin precursor (TN) (Human), transforming growth factor-beta-induced protein, and vimentin. 3. The method of claim 2, wherein the extracellular matrix coating further comprises type I collagen, type III collagen, fibronectin, decorin, biglycan, perlecan, and laminin. 4. The method of claim 3, wherein the extracellular matrix coating further comprises at least one of syndecan-1, collagen type V, or collagen type VI. 5-6. (canceled) 7. The method of claim 1, wherein the bone marrow stromal cells are isolated bone marrow mesenchymal stem cells. 8. The method of claim 1, wherein the mammalian mesenchymal stem cells are obtained from bone marrow or umbilical cord blood. 9. (canceled) 10. The method of the method of claim 1, wherein the microcarriers are spherical in shape. 11. The method of the method of claim 1, wherein the microcarriers have a positive charge. 12. (canceled) 13. The method of claim 11, wherein the microcarriers comprise are comprised of a cross-linked dextran matrix. 14. The method of the method of claim 1, wherein the microcarriers are cylindrical in shape. 15. The method of claim 14, wherein the microcarriers are hollow fibers. 16. (canceled) 17. The method of the method of claim 1, wherein the method further comprises culturing the bone marrow stromal cells under normoxic conditions. 18. The method of the method of claim 1, wherein the method further comprises culturing the mammalian mesenchymal stem cells under normoxic conditions. 19-22. (canceled) 23. A method of maintaining and expanding mammalian mesenchymal stem cells in culture in an undifferentiated state, the method comprising:
a. obtaining bone marrow stromal cell derived 3D extracellular matrix coated microcarriers; and b. culturing the mammalian mesenchymal stem cells in the presence of the extracellular matrix coated microcarriers, wherein the extracellular matrix coating restrains differentiation of the mammalian mesenchymal stem cells. 24. The method of claim 23, wherein the extracellular matrix coating comprises collagen alpha-1(XII), collagen alpha-3(VI), EMILIN-1, serpin H1, thrombospondin-1, tenascin precursor (TN) (Human), transforming growth factor-beta-induced protein, and vimentin. 25. The method of claim 24, wherein the extracellular matrix coating further comprises type I collagen, type III collagen, fibronectin, decorin, biglycan, perlecan, and laminin. 26. The method of claim 25, wherein the extracellular matrix coating further comprises at least one of syndecan-1, collagen type V, or collagen type VI. 27-44. (canceled) 45. A plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers. 46. The plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers of claim 45, further comprising mammalian mesenchymal stem cells attached to the plurality of extracellular matrix coated microcarriers. 47. (canceled) 48. The plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers of claim 45, wherein the extracellular matrix coated microcarriers are free or are substantially free of bone marrow stromal cells. 49. The plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers of claim 45, wherein the extracellular matrix coated microcarriers are comprised in a composition. 50. The plurality of bone marrow stromal cell derived 3D extracellular matrix coated microcarriers of claim 49, wherein the composition is a cell culture media. | 1,600 |
1,023 | 14,770,096 | 1,618 | Compositions comprising self-assembled hydrogel particles formed of short peptides which comprise one or more aromatic amino acid residue(s) in an inverted emulsion are disclosed. Such hydrogel particles which encapsulate an active agent and uses thereof in therapeutic and diagnostic applications are also disclosed. | 1. A composition comprising a plurality of physically discrete hydrogel particles, each hydrogel particle comprising a three-dimensional network made of a plurality of self-assembled peptides and an aqueous medium, wherein each peptide in the plurality of peptides comprises 2-6 amino acid residues, at least one of the amino acids being an aromatic amino acid. 2. (canceled) 3. The composition of claim 1, wherein an average diameter of each of the hydrogel particles ranges from 10 nm to 1000 nm, or from 10 nm to 500 nm. 4. The composition of claim 1, wherein the plurality of peptides comprises a plurality of dipeptides. 5. (canceled) 6. The composition of claim 4, wherein each peptide in the plurality of dipeptides is an aromatic-homodipeptide. 7. (canceled) 8. The composition of claim 6, wherein the plurality of aromatic dipeptides comprises a plurality of diphenylalanine peptides. 9. The composition of claim 1, wherein at least one of the peptides comprises a RGD sequence. 10. The composition of claim 1, wherein each peptide in the plurality of peptides comprises an RGD sequence. 11. The composition of claim 1, wherein at least one of the peptides comprises an end-capped moiety. 12. The composition of claim 11, wherein each of the peptides comprises an end-capped moiety. 13. The composition of claim 12, wherein the end-capping moiety is an aromatic end-capping moiety. 14-15. (canceled) 16. The composition of claim 1, wherein the plurality of peptides comprises a plurality of diphenylalanine peptides having an end-capping moiety substituting the N-terminus thereof. 17. The composition of claim 16, wherein each of the diphenylalanine peptide is an Fmoc-diphenylalanine (Fmoc-FF) peptide. 18. The composition of claim 1, wherein the plurality of peptides comprises a plurality of peptides having an FRGD sequence and an end-capping moiety substituting the N-terminus thereof. 19-21. (canceled) 22. The composition of claim 1, further comprising an emulsion stabilizer being in association with the particles. 23. The composition of claim 22, wherein the emulsion stabilizer is a Vitamin E derivative. 24-25. (canceled) 26. The composition of claim 1, wherein at least a portion of the hydrogel particles have a moiety or an agent incorporated therein and/or associated therewith. 27. (canceled) 28. The composition of claim 1, being is a form of a dry powder. 29. The composition of claim 28, further comprising a cryoprotectant. 30. The composition of claim 1, further comprising a pharmaceutically acceptable carrier. 31. (canceled) 32. A process of preparing the composition of claim 1, the process comprising:
adding an aqueous solution comprising the plurality of peptides to an organic solution, to thereby form an inverted (water-in-oil) emulsion; and subjecting the emulsion to high speed homogenization. 33. (canceled) 34. The process of claim 32, wherein the homogenization is performed at a speed rate of at least 10,000 rpm. 35. The process of claim 32, wherein the inverted emulsion further comprises an emulsion stabilizing agent. 36. The process of claim 35, wherein the emulsion stabilizing agent is a vitamin E derivative. 37. (canceled) 38. The process of claim 32, further comprising subjecting the hydrogel particles to lyophilization. 39. The process of claim 38, wherein the lyophilization is effected in the presence of a cryoprotecting agent. 40. The process of claim 32, wherein the hydrogel particles further comprise a bioactive agent incorporated therein, the process further comprising, prior to adding the aqueous solution of peptides to the organic solution, adding the bioactive agent to the aqueous solution of the peptides. 41. A method of delivering a bioactive agent to a bodily organ or tissue, the method comprising administering to the subject the composition of claim 26. 42. The method of claim 41, wherein the delivering is effected via systemic administration. 43. The method of claim 41, wherein the bioactive agent is a therapeutically active agent, the method being for treating a medical condition treatable by the bioactive agent. 44. The method of claim 41, wherein the bioactive agent is a diagnostic agent, the method being for monitoring a medical condition for which the diagnostic agent is indicative. | Compositions comprising self-assembled hydrogel particles formed of short peptides which comprise one or more aromatic amino acid residue(s) in an inverted emulsion are disclosed. Such hydrogel particles which encapsulate an active agent and uses thereof in therapeutic and diagnostic applications are also disclosed.1. A composition comprising a plurality of physically discrete hydrogel particles, each hydrogel particle comprising a three-dimensional network made of a plurality of self-assembled peptides and an aqueous medium, wherein each peptide in the plurality of peptides comprises 2-6 amino acid residues, at least one of the amino acids being an aromatic amino acid. 2. (canceled) 3. The composition of claim 1, wherein an average diameter of each of the hydrogel particles ranges from 10 nm to 1000 nm, or from 10 nm to 500 nm. 4. The composition of claim 1, wherein the plurality of peptides comprises a plurality of dipeptides. 5. (canceled) 6. The composition of claim 4, wherein each peptide in the plurality of dipeptides is an aromatic-homodipeptide. 7. (canceled) 8. The composition of claim 6, wherein the plurality of aromatic dipeptides comprises a plurality of diphenylalanine peptides. 9. The composition of claim 1, wherein at least one of the peptides comprises a RGD sequence. 10. The composition of claim 1, wherein each peptide in the plurality of peptides comprises an RGD sequence. 11. The composition of claim 1, wherein at least one of the peptides comprises an end-capped moiety. 12. The composition of claim 11, wherein each of the peptides comprises an end-capped moiety. 13. The composition of claim 12, wherein the end-capping moiety is an aromatic end-capping moiety. 14-15. (canceled) 16. The composition of claim 1, wherein the plurality of peptides comprises a plurality of diphenylalanine peptides having an end-capping moiety substituting the N-terminus thereof. 17. The composition of claim 16, wherein each of the diphenylalanine peptide is an Fmoc-diphenylalanine (Fmoc-FF) peptide. 18. The composition of claim 1, wherein the plurality of peptides comprises a plurality of peptides having an FRGD sequence and an end-capping moiety substituting the N-terminus thereof. 19-21. (canceled) 22. The composition of claim 1, further comprising an emulsion stabilizer being in association with the particles. 23. The composition of claim 22, wherein the emulsion stabilizer is a Vitamin E derivative. 24-25. (canceled) 26. The composition of claim 1, wherein at least a portion of the hydrogel particles have a moiety or an agent incorporated therein and/or associated therewith. 27. (canceled) 28. The composition of claim 1, being is a form of a dry powder. 29. The composition of claim 28, further comprising a cryoprotectant. 30. The composition of claim 1, further comprising a pharmaceutically acceptable carrier. 31. (canceled) 32. A process of preparing the composition of claim 1, the process comprising:
adding an aqueous solution comprising the plurality of peptides to an organic solution, to thereby form an inverted (water-in-oil) emulsion; and subjecting the emulsion to high speed homogenization. 33. (canceled) 34. The process of claim 32, wherein the homogenization is performed at a speed rate of at least 10,000 rpm. 35. The process of claim 32, wherein the inverted emulsion further comprises an emulsion stabilizing agent. 36. The process of claim 35, wherein the emulsion stabilizing agent is a vitamin E derivative. 37. (canceled) 38. The process of claim 32, further comprising subjecting the hydrogel particles to lyophilization. 39. The process of claim 38, wherein the lyophilization is effected in the presence of a cryoprotecting agent. 40. The process of claim 32, wherein the hydrogel particles further comprise a bioactive agent incorporated therein, the process further comprising, prior to adding the aqueous solution of peptides to the organic solution, adding the bioactive agent to the aqueous solution of the peptides. 41. A method of delivering a bioactive agent to a bodily organ or tissue, the method comprising administering to the subject the composition of claim 26. 42. The method of claim 41, wherein the delivering is effected via systemic administration. 43. The method of claim 41, wherein the bioactive agent is a therapeutically active agent, the method being for treating a medical condition treatable by the bioactive agent. 44. The method of claim 41, wherein the bioactive agent is a diagnostic agent, the method being for monitoring a medical condition for which the diagnostic agent is indicative. | 1,600 |
1,024 | 16,201,410 | 1,628 | Provided herein are methods for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is also being administered an organic anion transporter-1 (OAT1) substrate. The method comprises administering to the subject a therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, and monitoring the subject for signs and symptoms of toxicity and clinical response associated with the OAT1 substrate. | 1. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need of dichlorphenamide, or a pharmaceutically acceptable salt thereof to treat a first disease or disorder, wherein the subject is being administered an organic anion transporter-1 (OAT1) substrate for the treatment of a second disease or disorder, the method comprising:
discontinuing administration of the OAT1 substrate, and administering to the subject an initial dose of dichlorphenamide, or a pharmaceutically acceptable salt thereof, of 50 mg twice daily. 2. (canceled) 3. (canceled) 4. (canceled) 5.-23. (canceled) 24. (canceled) 25. The method of claim 1, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises the up-titration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, at weekly intervals until a modified dose is administered. 26. The method of claim 25, wherein the modified dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 200 mg. 27. The method of claim 1, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises administering a first dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, for a period of about one week;
further increasing the dose by an amount equal to an incremental value; and determining whether the subject tolerates the further increased dose; wherein the cycle is repeated so long as the subject tolerates the further increased dose, wherein the incremental value at each cycle repetition is the same or different; and wherein if the subject does not tolerate the further increased dose, the modified dose for the subject is equal to the difference between the further increased dose and the incremental value for the last cycle repetition. 28. (canceled) 29. (canceled) 30. (canceled) 31. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, the method comprising:
administering to the subject between 25 mg and 200 mg per day of dichlorphenamide, or a pharmaceutically acceptable salt thereof, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof is administered in the absence of an OAT1 substrate. 32. (canceled) 33. (canceled) 34. The method of claim 31, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg twice daily. 35. The method of claim 31, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises the up-titration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, at weekly intervals until a modified dose is administered. 36. The method of claim 35, wherein the modified dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 200 mg. 37. The method of claim 31, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises administering a first dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, for a period of about one week;
further increasing the dose by an amount equal to an incremental value; and determining whether the subject tolerates the further increased dose; wherein the cycle is repeated so long as the subject tolerates the further increased dose, wherein the incremental value at each cycle repetition is the same or different; and wherein if the subject does not tolerate the further increased dose, the modified dose for the subject is equal to the difference between the further increased dose and the incremental value for the last cycle repetition. 38. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need of dichlorphenamide, or a pharmaceutically acceptable salt thereof to treat a first disease or disorder, wherein the subject is being administered an organic anion transporter-1 (OAT1) substrate for the treatment of a second disease or disorder, the method comprising:
discontinuing administration of the OAT1 substrate, and administering to the subject an initial dose of dichlorphenamide, or a pharmaceutically acceptable salt thereof, of 50 mg once daily. 39. (canceled) 40. (canceled) 41. The method of claim 38, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises the up-titration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, at weekly intervals until a modified dose is administered. 42. The method of claim 41, wherein the modified dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 200 mg. 43. The method of claim 38, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises administering a first dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, for a period of about one week;
further increasing the dose by an amount equal to an incremental value; and determining whether the subject tolerates the further increased dose; wherein the cycle is repeated so long as the subject tolerates the further increased dose, wherein the incremental value at each cycle repetition is the same or different; and wherein if the subject does not tolerate the further increased dose, the modified dose for the subject is equal to the difference between the further increased dose and the incremental value for the last cycle repetition. | Provided herein are methods for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is also being administered an organic anion transporter-1 (OAT1) substrate. The method comprises administering to the subject a therapeutically effective amount of dichlorphenamide, or a pharmaceutically acceptable salt thereof, and monitoring the subject for signs and symptoms of toxicity and clinical response associated with the OAT1 substrate.1. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need of dichlorphenamide, or a pharmaceutically acceptable salt thereof to treat a first disease or disorder, wherein the subject is being administered an organic anion transporter-1 (OAT1) substrate for the treatment of a second disease or disorder, the method comprising:
discontinuing administration of the OAT1 substrate, and administering to the subject an initial dose of dichlorphenamide, or a pharmaceutically acceptable salt thereof, of 50 mg twice daily. 2. (canceled) 3. (canceled) 4. (canceled) 5.-23. (canceled) 24. (canceled) 25. The method of claim 1, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises the up-titration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, at weekly intervals until a modified dose is administered. 26. The method of claim 25, wherein the modified dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 200 mg. 27. The method of claim 1, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises administering a first dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, for a period of about one week;
further increasing the dose by an amount equal to an incremental value; and determining whether the subject tolerates the further increased dose; wherein the cycle is repeated so long as the subject tolerates the further increased dose, wherein the incremental value at each cycle repetition is the same or different; and wherein if the subject does not tolerate the further increased dose, the modified dose for the subject is equal to the difference between the further increased dose and the incremental value for the last cycle repetition. 28. (canceled) 29. (canceled) 30. (canceled) 31. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, the method comprising:
administering to the subject between 25 mg and 200 mg per day of dichlorphenamide, or a pharmaceutically acceptable salt thereof, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof is administered in the absence of an OAT1 substrate. 32. (canceled) 33. (canceled) 34. The method of claim 31, wherein the therapeutically effective amount of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 50 mg twice daily. 35. The method of claim 31, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises the up-titration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, at weekly intervals until a modified dose is administered. 36. The method of claim 35, wherein the modified dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 200 mg. 37. The method of claim 31, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises administering a first dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, for a period of about one week;
further increasing the dose by an amount equal to an incremental value; and determining whether the subject tolerates the further increased dose; wherein the cycle is repeated so long as the subject tolerates the further increased dose, wherein the incremental value at each cycle repetition is the same or different; and wherein if the subject does not tolerate the further increased dose, the modified dose for the subject is equal to the difference between the further increased dose and the incremental value for the last cycle repetition. 38. A method for administering dichlorphenamide, or a pharmaceutically acceptable salt thereof, to a subject in need of dichlorphenamide, or a pharmaceutically acceptable salt thereof to treat a first disease or disorder, wherein the subject is being administered an organic anion transporter-1 (OAT1) substrate for the treatment of a second disease or disorder, the method comprising:
discontinuing administration of the OAT1 substrate, and administering to the subject an initial dose of dichlorphenamide, or a pharmaceutically acceptable salt thereof, of 50 mg once daily. 39. (canceled) 40. (canceled) 41. The method of claim 38, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises the up-titration of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, at weekly intervals until a modified dose is administered. 42. The method of claim 41, wherein the modified dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is 200 mg. 43. The method of claim 38, wherein the dichlorphenamide, or a pharmaceutically acceptable salt thereof, is administered via a titration scheme that comprises administering a first dose of the dichlorphenamide, or a pharmaceutically acceptable salt thereof, for a period of about one week;
further increasing the dose by an amount equal to an incremental value; and determining whether the subject tolerates the further increased dose; wherein the cycle is repeated so long as the subject tolerates the further increased dose, wherein the incremental value at each cycle repetition is the same or different; and wherein if the subject does not tolerate the further increased dose, the modified dose for the subject is equal to the difference between the further increased dose and the incremental value for the last cycle repetition. | 1,600 |
1,025 | 15,294,488 | 1,633 | Non-human animals, tissues, cells, and genetic material are provided that comprise a modification of an endogenous non-human heavy chain immunoglobulin sequence and that comprise an ADAM6 activity functional in a rodent (e.g., a mouse), wherein the non-human animals rearrange human immunoglobulin light chain gene segments in the context of heavy chain constant regions and express immunoglobulin-like molecules comprising human immunoglobulin light chain variable domains fused to heavy chain constant domains that are cognate with human immunoglobulin light chain variable domains fused to light chain constant domains. | 1-35. (canceled) 36. A method for making a human antigen binding protein, comprising:
culturing a host cell comprising a first nucleotide sequence including a first human light chain variable region sequence operably linked to a human heavy chain constant region sequence and a second nucleotide sequence including a second human light chain variable region sequence operably linked to a human light chain constant region sequence, so that a human antigen binding protein is expressed from the first and second nucleotide sequences, wherein the first and second human light chain variable region sequences were identified from a cell of a non-human animal that includes in its genome:
(i) an insertion of one or more human VL gene segments and one or more human JL gene segments upstream of a non-human light chain constant region sequence, wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to the non-human light chain constant region sequence,
(ii) an insertion of one or more human VL gene segments and one or more human JL gene segments upstream of a non-human heavy chain constant region sequence, wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to the non-human heavy chain constant region sequence, and
(iii) an inserted nucleic acid sequence that encodes a rodent ADAM6 protein, wherein the rodent ADAM6 protein is expressed from the inserted nucleic acid sequence. 37. The method of claim 36, wherein the method further comprises:
obtaining the first human light chain variable region sequence from a nucleotide sequence that encodes a first human light chain variable domain fused to a non-human heavy chain constant region found in a cell of the non-human animal, and the second human light chain variable region sequence from a nucleotide sequence that encodes a second human light chain variable domain fused to a non-human light chain constant region found in a cell of the non-human animal. 38. The method of claim 37, wherein the method further comprises:
cloning the first human light chain variable region sequence in frame with a human heavy chain constant region and the second human light chain variable region sequence in frame with a human light chain constant region. 39. The method of claim 36, wherein the method further comprises:
(a) exposing the non-human animal to an antigen of interest, (b) isolating one or more B lymphocytes of the non-human animal, wherein the one or more B lymphocytes express an antibody that includes the first human light chain variable domain encoded by the first human light chain variable region sequence and the second human light chain variable domain encoded by the second human light chain variable region sequence. 40. The method of claim 36, wherein the non-human light chain constant region sequence of (i) is a rodent light chain constant region sequence and/or the non-human heavy chain constant region sequence of (ii) is a rodent heavy chain constant region sequence. 41. The method of claim 36, wherein the non-human light chain constant region sequence of (i) is a mouse Cκ and the non-human heavy chain constant region sequence of (ii) is a mouse heavy chain constant region sequence. 42. The method of claim 36, wherein the one or more human VL gene segments and the one or more human JL gene segments upstream of the non-human light chain constant region sequence of (i) are one or more human Vκ gene segments and one or more human Jκ gene segments. 43. The method of claim 36, wherein the one or more human VL gene segments and the one or more human JL gene segments upstream of the non-human heavy chain constant region sequence of (ii) are one or more human Vκ gene segments and one or more human Jκ gene segments. 44. The method of claim 36, wherein
(a) the inserted nucleic acid that encodes the non-human ADAM6 protein is present between one of the one or more human VL gene segments and one of the one or more human JL gene segments of (ii); (b) the non-human animal includes two or more human VL gene segments and the inserted nucleic acid that encodes the non-human ADAM6 protein is present between two of the two or more human VL gene segments; or (c) the non-human animal includes two or more human JL gene segments and the inserted nucleic acid that encodes the non-human ADAM6 protein is present between two of the two or more human JL gene segments. 45. The method of claim 36, wherein the non-human animal comprises one or more endogenous VL gene segments and/or one or more endogenous JL gene segments that are incapable of rearranging to form an immunoglobulin light chain variable region sequence in the non-human animal. 46. The method of claim 36, wherein the host cell is selected from a B cell, a hybridoma, a quadroma, a CHO cell, a COS cell, a 293 cell, a Hela cell, and a human retinal cell expressing a viral nucleic acid sequence. 47. The method of claim 46, wherein the human heavy chain constant region of (a) comprises a human IgG isotype. 48. The method of claim 47, wherein the human IgG isotype is selected from an IgG1, IgG2, and IgG4. 49. The method of claim 47, wherein the human IgG4 is a modified IgG4. 50. The method of claim 49, wherein the modified IgG4 comprises a substitution in the hinge region. 51. The method of claim 36, wherein the cell of the non-human animal is a lymphocyte or splenocyte. 52. A method of making a nucleic acid that encodes an immunoglobulin light chain variable domain comprising:
(a) immunizing a non-human animal with an antigen of interest, wherein the genome of the non-human animal comprises:
(i) an insertion of one or more human VL gene segments and one or more human JL gene segments upstream of a non-human light chain constant region sequence, wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to the non-human light chain constant region sequence,
(ii) an insertion of one or more human VL gene segments and one or more human JL gene segments upstream of a non-human heavy chain constant region sequence, wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to the non-human heavy chain constant region sequence, and
(iii) an inserted nucleic acid sequence that encodes a rodent ADAM6 protein, wherein the rodent ADAM6 protein is expressed from the inserted nucleic acid sequence,
(b) isolating one or more B lymphocytes of the non-human animal, wherein the one or more B lymphocytes express an antibody that binds the antigen of interest, and (c) identifying a nucleic acid sequence that encodes a light chain variable domain of the antibody of (b) that binds that antigen of interest, wherein the antibody of (b) comprises:
(A) two light chains, wherein each light chain comprises a human light chain variable domain and a non-human light chain constant domain, and
(B) two heavy chains, wherein each heavy chain comprises a human light chain variable domain and a non-human heavy chain constant domain. 53. The method of claim 52, wherein the non-human light chain constant region sequence of (i) is a rodent light chain constant region sequence and/or the non-human heavy chain constant region sequence of (ii) is a rodent heavy chain constant region sequence. 54. The method of claim 52, wherein the non-human light chain constant region sequence of (i) is a mouse Cκ and the non-human heavy chain constant region sequence of (ii) is a mouse heavy chain constant region. 55. The method of claim 52, wherein the one or more human VL gene segments and the one or more human JL gene segments upstream of the non-human immunoglobulin light chain constant region sequence of (i) are one or more human Vκ gene segments and one or more human Jκ gene segments. 56. The method of claim 52, wherein the one or more human VL gene segments and the one or more human JL gene segments upstream of the non-human immunoglobulin heavy chain constant region sequence of (ii) are one or more human Vκ gene segments and one or more human Jκ gene segments. 57. The method of claim 52, wherein
(a) the inserted nucleic acid that encodes the rodent ADAM6 protein is present between one of the one or more human VL gene segments and one of the one or more human JL gene segments of (ii); (b) the non-human animal includes two or more human VL gene segments and the inserted nucleic acid that encodes the rodent ADAM6 protein is present between two of the two or more human VL gene segments; or (c) the non-human animal includes two or more human JL gene segments and the inserted nucleic acid that encodes the rodent ADAM6 protein is present between two of the two or more human JL gene segments. 58. The method of claim 52, wherein the non-human animal comprises one or more endogenous VL gene segments and/or one or more endogenous JL gene segments that are incapable of rearranging to form an immunoglobulin light chain variable region sequence in the non-human animal. | Non-human animals, tissues, cells, and genetic material are provided that comprise a modification of an endogenous non-human heavy chain immunoglobulin sequence and that comprise an ADAM6 activity functional in a rodent (e.g., a mouse), wherein the non-human animals rearrange human immunoglobulin light chain gene segments in the context of heavy chain constant regions and express immunoglobulin-like molecules comprising human immunoglobulin light chain variable domains fused to heavy chain constant domains that are cognate with human immunoglobulin light chain variable domains fused to light chain constant domains.1-35. (canceled) 36. A method for making a human antigen binding protein, comprising:
culturing a host cell comprising a first nucleotide sequence including a first human light chain variable region sequence operably linked to a human heavy chain constant region sequence and a second nucleotide sequence including a second human light chain variable region sequence operably linked to a human light chain constant region sequence, so that a human antigen binding protein is expressed from the first and second nucleotide sequences, wherein the first and second human light chain variable region sequences were identified from a cell of a non-human animal that includes in its genome:
(i) an insertion of one or more human VL gene segments and one or more human JL gene segments upstream of a non-human light chain constant region sequence, wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to the non-human light chain constant region sequence,
(ii) an insertion of one or more human VL gene segments and one or more human JL gene segments upstream of a non-human heavy chain constant region sequence, wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to the non-human heavy chain constant region sequence, and
(iii) an inserted nucleic acid sequence that encodes a rodent ADAM6 protein, wherein the rodent ADAM6 protein is expressed from the inserted nucleic acid sequence. 37. The method of claim 36, wherein the method further comprises:
obtaining the first human light chain variable region sequence from a nucleotide sequence that encodes a first human light chain variable domain fused to a non-human heavy chain constant region found in a cell of the non-human animal, and the second human light chain variable region sequence from a nucleotide sequence that encodes a second human light chain variable domain fused to a non-human light chain constant region found in a cell of the non-human animal. 38. The method of claim 37, wherein the method further comprises:
cloning the first human light chain variable region sequence in frame with a human heavy chain constant region and the second human light chain variable region sequence in frame with a human light chain constant region. 39. The method of claim 36, wherein the method further comprises:
(a) exposing the non-human animal to an antigen of interest, (b) isolating one or more B lymphocytes of the non-human animal, wherein the one or more B lymphocytes express an antibody that includes the first human light chain variable domain encoded by the first human light chain variable region sequence and the second human light chain variable domain encoded by the second human light chain variable region sequence. 40. The method of claim 36, wherein the non-human light chain constant region sequence of (i) is a rodent light chain constant region sequence and/or the non-human heavy chain constant region sequence of (ii) is a rodent heavy chain constant region sequence. 41. The method of claim 36, wherein the non-human light chain constant region sequence of (i) is a mouse Cκ and the non-human heavy chain constant region sequence of (ii) is a mouse heavy chain constant region sequence. 42. The method of claim 36, wherein the one or more human VL gene segments and the one or more human JL gene segments upstream of the non-human light chain constant region sequence of (i) are one or more human Vκ gene segments and one or more human Jκ gene segments. 43. The method of claim 36, wherein the one or more human VL gene segments and the one or more human JL gene segments upstream of the non-human heavy chain constant region sequence of (ii) are one or more human Vκ gene segments and one or more human Jκ gene segments. 44. The method of claim 36, wherein
(a) the inserted nucleic acid that encodes the non-human ADAM6 protein is present between one of the one or more human VL gene segments and one of the one or more human JL gene segments of (ii); (b) the non-human animal includes two or more human VL gene segments and the inserted nucleic acid that encodes the non-human ADAM6 protein is present between two of the two or more human VL gene segments; or (c) the non-human animal includes two or more human JL gene segments and the inserted nucleic acid that encodes the non-human ADAM6 protein is present between two of the two or more human JL gene segments. 45. The method of claim 36, wherein the non-human animal comprises one or more endogenous VL gene segments and/or one or more endogenous JL gene segments that are incapable of rearranging to form an immunoglobulin light chain variable region sequence in the non-human animal. 46. The method of claim 36, wherein the host cell is selected from a B cell, a hybridoma, a quadroma, a CHO cell, a COS cell, a 293 cell, a Hela cell, and a human retinal cell expressing a viral nucleic acid sequence. 47. The method of claim 46, wherein the human heavy chain constant region of (a) comprises a human IgG isotype. 48. The method of claim 47, wherein the human IgG isotype is selected from an IgG1, IgG2, and IgG4. 49. The method of claim 47, wherein the human IgG4 is a modified IgG4. 50. The method of claim 49, wherein the modified IgG4 comprises a substitution in the hinge region. 51. The method of claim 36, wherein the cell of the non-human animal is a lymphocyte or splenocyte. 52. A method of making a nucleic acid that encodes an immunoglobulin light chain variable domain comprising:
(a) immunizing a non-human animal with an antigen of interest, wherein the genome of the non-human animal comprises:
(i) an insertion of one or more human VL gene segments and one or more human JL gene segments upstream of a non-human light chain constant region sequence, wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to the non-human light chain constant region sequence,
(ii) an insertion of one or more human VL gene segments and one or more human JL gene segments upstream of a non-human heavy chain constant region sequence, wherein the one or more human VL gene segments and one or more human JL gene segments are operably linked to the non-human heavy chain constant region sequence, and
(iii) an inserted nucleic acid sequence that encodes a rodent ADAM6 protein, wherein the rodent ADAM6 protein is expressed from the inserted nucleic acid sequence,
(b) isolating one or more B lymphocytes of the non-human animal, wherein the one or more B lymphocytes express an antibody that binds the antigen of interest, and (c) identifying a nucleic acid sequence that encodes a light chain variable domain of the antibody of (b) that binds that antigen of interest, wherein the antibody of (b) comprises:
(A) two light chains, wherein each light chain comprises a human light chain variable domain and a non-human light chain constant domain, and
(B) two heavy chains, wherein each heavy chain comprises a human light chain variable domain and a non-human heavy chain constant domain. 53. The method of claim 52, wherein the non-human light chain constant region sequence of (i) is a rodent light chain constant region sequence and/or the non-human heavy chain constant region sequence of (ii) is a rodent heavy chain constant region sequence. 54. The method of claim 52, wherein the non-human light chain constant region sequence of (i) is a mouse Cκ and the non-human heavy chain constant region sequence of (ii) is a mouse heavy chain constant region. 55. The method of claim 52, wherein the one or more human VL gene segments and the one or more human JL gene segments upstream of the non-human immunoglobulin light chain constant region sequence of (i) are one or more human Vκ gene segments and one or more human Jκ gene segments. 56. The method of claim 52, wherein the one or more human VL gene segments and the one or more human JL gene segments upstream of the non-human immunoglobulin heavy chain constant region sequence of (ii) are one or more human Vκ gene segments and one or more human Jκ gene segments. 57. The method of claim 52, wherein
(a) the inserted nucleic acid that encodes the rodent ADAM6 protein is present between one of the one or more human VL gene segments and one of the one or more human JL gene segments of (ii); (b) the non-human animal includes two or more human VL gene segments and the inserted nucleic acid that encodes the rodent ADAM6 protein is present between two of the two or more human VL gene segments; or (c) the non-human animal includes two or more human JL gene segments and the inserted nucleic acid that encodes the rodent ADAM6 protein is present between two of the two or more human JL gene segments. 58. The method of claim 52, wherein the non-human animal comprises one or more endogenous VL gene segments and/or one or more endogenous JL gene segments that are incapable of rearranging to form an immunoglobulin light chain variable region sequence in the non-human animal. | 1,600 |
1,026 | 15,645,436 | 1,616 | The invention relates to a solid pharmaceutical preparation comprising levothyroxine sodium, gelatine, citric acid and a filler. The solid pharmaceutical preparation has an improved stability. | 1. Solid pharmaceutical preparation comprising levothyroxine sodium, gelatine, citric acid and a filler. 2. Solid pharmaceutical preparation according to claim 1, characterized in that it comprises liothyronine sodium. 3. Solid pharmaceutical preparation according to claim 1, characterized in that the filler is a sugar alcohol such as sorbitol or mannitol dulcitol, xylitol or ribitol, preferably sorbitol or mannitol, particular preferably mannitol, a sugar such as glucose, fructose, mannose, lactose, saccharose or maltose, preferably lactose, saccharose or maltose, particular preferably lactose, a starch such as potato starch, rice starch, maize starch or pregelatinized starch, preferably maize starch or pregelatinized starch, particular preferably maize starch, a cellulose such as powdered cellulose or microcrystalline cellulose, preferably microcrystalline cellulose, or a mixture thereof. 4. Solid pharmaceutical preparation according to claim 3, characterised in that the filler is mannitol and/or maize starch. 5. Solid pharmaceutical preparation according to claim 1, characterized in that it further comprises an antioxidant selected from the group consisting of tocopherol, propyl gallate, tertiary butylhydroquinone, butylated hydroxyanisole and butylated hydroxytoluene, preferably hydroxyanisole or butylated hydroxytoluene, particular preferably butylated hydroxytoluene. 6. Solid pharmaceutical preparation according to claim 1, characterised in that it is in granule, pellet, capsule or tablet form. 7. Solid pharmaceutical preparation according to claim 6, characterised in that it is a tablet. 8. Solid pharmaceutical preparation according to claim 1, characterised in that at least one disintegrating agent is present. 9. Solid pharmaceutical preparation according to claim 8, characterized in that the disintegrating agent is sodium starch glycolate, or carboxymethylcellulose sodium or a mixture thereof. 10. Solid pharmaceutical preparation according to claim 9, characterized in that the disintegrating agent present is carboxymethylcellulose sodium. 11. Solid pharmaceutical preparation according to claim 1, characterised in that it comprises 1 to 10% by weight of gelatine, 0.1 to 3% by weight citric acid, 50 to 80% by weight of mannitol or lactose, 10 to 30% by weight maize starch. 12. Solid pharmaceutical preparation according to claim 11, characterised in that it comprises 0,05 to 0,5% by weight butylated hydroxytoluene. 13. Process for the production of a solid pharmaceutical preparation according to claim 7, characterized in that
(a) levothyroxine sodium and optionally liothyronine sodium is/are suspended in an aqueous gelatine solution, (b) the suspension obtained by step (a) is sprayed onto the filler in a fluidized bed granulation and dried to form granules, (c) the granules obtained by step (b) is collected and optionally, (d) a disintegrant and optionally a lubricant is/are mixed with the granules obtained by step (c), and (e) the mixture obtained by step (d) is compressed to give tablets. 14. Process for the production of a solid pharmaceutical preparation according to 13. , characterized in that citric acid and, if present, the antioxidant is dissolved in the aqueous gelatine solution used in step (a) or is admixed with the granules in step (d). 15. Process for the preparation of a solid pharmaceutical preparation according to claim 13, characterised in that the granules or the tablets are provided with a coating. 16. A process for the stabilization of levothyroxine sodium, comprising adding to said levothyroxine sodium a stabilizing-effective amount of gelatine, citric acid and a filler. 17. The process according to claim 16, wherein the filler is dulcitol, xylitol, ribitol, sorbitol, mannitol, glucose, fructose, mannose, lactose, saccharose, maltose, potato starch, rice starch, maize starch, pregelatinized starch, powdered cellulose, microcrystalline cellulose or a mixture thereof. | The invention relates to a solid pharmaceutical preparation comprising levothyroxine sodium, gelatine, citric acid and a filler. The solid pharmaceutical preparation has an improved stability.1. Solid pharmaceutical preparation comprising levothyroxine sodium, gelatine, citric acid and a filler. 2. Solid pharmaceutical preparation according to claim 1, characterized in that it comprises liothyronine sodium. 3. Solid pharmaceutical preparation according to claim 1, characterized in that the filler is a sugar alcohol such as sorbitol or mannitol dulcitol, xylitol or ribitol, preferably sorbitol or mannitol, particular preferably mannitol, a sugar such as glucose, fructose, mannose, lactose, saccharose or maltose, preferably lactose, saccharose or maltose, particular preferably lactose, a starch such as potato starch, rice starch, maize starch or pregelatinized starch, preferably maize starch or pregelatinized starch, particular preferably maize starch, a cellulose such as powdered cellulose or microcrystalline cellulose, preferably microcrystalline cellulose, or a mixture thereof. 4. Solid pharmaceutical preparation according to claim 3, characterised in that the filler is mannitol and/or maize starch. 5. Solid pharmaceutical preparation according to claim 1, characterized in that it further comprises an antioxidant selected from the group consisting of tocopherol, propyl gallate, tertiary butylhydroquinone, butylated hydroxyanisole and butylated hydroxytoluene, preferably hydroxyanisole or butylated hydroxytoluene, particular preferably butylated hydroxytoluene. 6. Solid pharmaceutical preparation according to claim 1, characterised in that it is in granule, pellet, capsule or tablet form. 7. Solid pharmaceutical preparation according to claim 6, characterised in that it is a tablet. 8. Solid pharmaceutical preparation according to claim 1, characterised in that at least one disintegrating agent is present. 9. Solid pharmaceutical preparation according to claim 8, characterized in that the disintegrating agent is sodium starch glycolate, or carboxymethylcellulose sodium or a mixture thereof. 10. Solid pharmaceutical preparation according to claim 9, characterized in that the disintegrating agent present is carboxymethylcellulose sodium. 11. Solid pharmaceutical preparation according to claim 1, characterised in that it comprises 1 to 10% by weight of gelatine, 0.1 to 3% by weight citric acid, 50 to 80% by weight of mannitol or lactose, 10 to 30% by weight maize starch. 12. Solid pharmaceutical preparation according to claim 11, characterised in that it comprises 0,05 to 0,5% by weight butylated hydroxytoluene. 13. Process for the production of a solid pharmaceutical preparation according to claim 7, characterized in that
(a) levothyroxine sodium and optionally liothyronine sodium is/are suspended in an aqueous gelatine solution, (b) the suspension obtained by step (a) is sprayed onto the filler in a fluidized bed granulation and dried to form granules, (c) the granules obtained by step (b) is collected and optionally, (d) a disintegrant and optionally a lubricant is/are mixed with the granules obtained by step (c), and (e) the mixture obtained by step (d) is compressed to give tablets. 14. Process for the production of a solid pharmaceutical preparation according to 13. , characterized in that citric acid and, if present, the antioxidant is dissolved in the aqueous gelatine solution used in step (a) or is admixed with the granules in step (d). 15. Process for the preparation of a solid pharmaceutical preparation according to claim 13, characterised in that the granules or the tablets are provided with a coating. 16. A process for the stabilization of levothyroxine sodium, comprising adding to said levothyroxine sodium a stabilizing-effective amount of gelatine, citric acid and a filler. 17. The process according to claim 16, wherein the filler is dulcitol, xylitol, ribitol, sorbitol, mannitol, glucose, fructose, mannose, lactose, saccharose, maltose, potato starch, rice starch, maize starch, pregelatinized starch, powdered cellulose, microcrystalline cellulose or a mixture thereof. | 1,600 |
1,027 | 15,054,803 | 1,658 | A beverage comprising protein, plant sterol ester and/or plant stanol ester, a fruit and/or vegetable preparation, and a stabiliser. | 1. A beverage comprising:
protein in an amount of 0.5-5% by weight of the beverage; plant sterol ester and/or plant stanol ester in an amount of 0.5-7% by weight of the beverage; stabilizer in an amount of 0.1-1% by weight of the beverage; and a vegetable preparation, wherein the beverage has an effective concentration of 30-1000% by weight natural vegetable derived from the vegetable preparation, based on the weight of the beverage. 2. The beverage of claim 1, wherein the amount of plant sterol ester and/or plant stanol ester is from 1.0 to 4.0% by weight of the beverage. 3. The beverage of claim 1, wherein the vegetable preparation comprises at least one of carrot, tomato, maize, celery, cucumber, beets, parsley, cabbage, lettuce, spinach, wheat grass, pea, pumpkin, or sauerkraut. 4. The beverage of claim 1, wherein the protein is derived from dairy milk, soy milk, rice milk, or a mixture thereof. 5. The beverage of claim 1, wherein the amount of protein is from 1.0 to 3.0% by weight of the beverage. 6. The beverage of claim 1, wherein the amount of protein is from 0.75 to 2.5% by weight of the beverage. 7. The beverage of claim 1, wherein the beverage further comprises an emulsifier. 8. The beverage of claim 7, wherein the emulsifier is selected from the group consisting of acetic acid esters, lactic acid esters, succinic acid esters, citric acid esters, diacetyl tartaric acid esters and diacetyl lactic acid esters of mono- and diglycerides, sorbitan esters, polysorbates, stearoyl lactylates, and sugar esters. 9. The beverage of claim 7, wherein the emulsifier is present in an amount of 0.05-0.5% by weight of the beverage. 10. The beverage of claim 1, wherein the amount of vegetable preparation is from 11.5% to 79.35% by weight of the beverage. 11. The beverage of claim 1, wherein the effective concentration of natural vegetable derived from the vegetable preparation is from 40 to 750% by weight of the beverage. 12. The beverage of claim 1, wherein the effective concentration of natural vegetable derived from the vegetable preparation is at least 101% by weight of the beverage. 13. The beverage of claim 1, wherein the amount of stabilizer is from 0.2 to 0.8% by weight of the beverage. 14. The beverage of claim 1, wherein at least one stabilizer is selected from the group consisting of pectin, modified starch, xanthan gum, and guar gum. 15. The beverage of claim 1, wherein the stabilizer comprises pectin. 16. The beverage of claim 1, wherein the stabilizer comprises high methoxy pectin. 17. The beverage of claim 1, wherein the protein comprises non-fermented protein. 18. The beverage of claim 1, wherein the beverage is free of a carbohydrate sweetening agent. 19. The beverage of claim 1, wherein the beverage further comprises a non-carbohydrate sweetener. 20. The beverage of claim 1, wherein the ingredients are homogenously dispersed throughout the beverage. 21. The beverage of claim 1, wherein at least 90% of particles present in the beverage have a particle size less than 100 μm. 22. A beverage comprising:
protein in an amount of 0.5-5% by weight of the beverage; plant sterol ester and/or plant stanol ester in an amount of 0.5-7% by weight of the beverage; stabilizer in an amount of 0.1-1% by weight of the beverage, wherein the stabilizer comprises high methoxy pectin; and a fruit and/or vegetable preparation, wherein the beverage has an effective concentration of 30-1000% by weight natural fruit and/or vegetable derived from the fruit and/or vegetable preparation, based on the weight of the beverage. 23. A beverage comprising:
dairy milk, soy milk, rice milk, or a mixture thereof in an amount of 20-80% by weight of the beverage; plant sterol ester and/or plant stanol ester in an amount of 0.5-7% by weight of the beverage; stabilizer in an amount of 0.1-1% by weight of the beverage, wherein the stabilizer is high methoxy pectin; and a fruit and/or vegetable preparation in an amount of 23.5 to 74.25% by weight of the beverage. | A beverage comprising protein, plant sterol ester and/or plant stanol ester, a fruit and/or vegetable preparation, and a stabiliser.1. A beverage comprising:
protein in an amount of 0.5-5% by weight of the beverage; plant sterol ester and/or plant stanol ester in an amount of 0.5-7% by weight of the beverage; stabilizer in an amount of 0.1-1% by weight of the beverage; and a vegetable preparation, wherein the beverage has an effective concentration of 30-1000% by weight natural vegetable derived from the vegetable preparation, based on the weight of the beverage. 2. The beverage of claim 1, wherein the amount of plant sterol ester and/or plant stanol ester is from 1.0 to 4.0% by weight of the beverage. 3. The beverage of claim 1, wherein the vegetable preparation comprises at least one of carrot, tomato, maize, celery, cucumber, beets, parsley, cabbage, lettuce, spinach, wheat grass, pea, pumpkin, or sauerkraut. 4. The beverage of claim 1, wherein the protein is derived from dairy milk, soy milk, rice milk, or a mixture thereof. 5. The beverage of claim 1, wherein the amount of protein is from 1.0 to 3.0% by weight of the beverage. 6. The beverage of claim 1, wherein the amount of protein is from 0.75 to 2.5% by weight of the beverage. 7. The beverage of claim 1, wherein the beverage further comprises an emulsifier. 8. The beverage of claim 7, wherein the emulsifier is selected from the group consisting of acetic acid esters, lactic acid esters, succinic acid esters, citric acid esters, diacetyl tartaric acid esters and diacetyl lactic acid esters of mono- and diglycerides, sorbitan esters, polysorbates, stearoyl lactylates, and sugar esters. 9. The beverage of claim 7, wherein the emulsifier is present in an amount of 0.05-0.5% by weight of the beverage. 10. The beverage of claim 1, wherein the amount of vegetable preparation is from 11.5% to 79.35% by weight of the beverage. 11. The beverage of claim 1, wherein the effective concentration of natural vegetable derived from the vegetable preparation is from 40 to 750% by weight of the beverage. 12. The beverage of claim 1, wherein the effective concentration of natural vegetable derived from the vegetable preparation is at least 101% by weight of the beverage. 13. The beverage of claim 1, wherein the amount of stabilizer is from 0.2 to 0.8% by weight of the beverage. 14. The beverage of claim 1, wherein at least one stabilizer is selected from the group consisting of pectin, modified starch, xanthan gum, and guar gum. 15. The beverage of claim 1, wherein the stabilizer comprises pectin. 16. The beverage of claim 1, wherein the stabilizer comprises high methoxy pectin. 17. The beverage of claim 1, wherein the protein comprises non-fermented protein. 18. The beverage of claim 1, wherein the beverage is free of a carbohydrate sweetening agent. 19. The beverage of claim 1, wherein the beverage further comprises a non-carbohydrate sweetener. 20. The beverage of claim 1, wherein the ingredients are homogenously dispersed throughout the beverage. 21. The beverage of claim 1, wherein at least 90% of particles present in the beverage have a particle size less than 100 μm. 22. A beverage comprising:
protein in an amount of 0.5-5% by weight of the beverage; plant sterol ester and/or plant stanol ester in an amount of 0.5-7% by weight of the beverage; stabilizer in an amount of 0.1-1% by weight of the beverage, wherein the stabilizer comprises high methoxy pectin; and a fruit and/or vegetable preparation, wherein the beverage has an effective concentration of 30-1000% by weight natural fruit and/or vegetable derived from the fruit and/or vegetable preparation, based on the weight of the beverage. 23. A beverage comprising:
dairy milk, soy milk, rice milk, or a mixture thereof in an amount of 20-80% by weight of the beverage; plant sterol ester and/or plant stanol ester in an amount of 0.5-7% by weight of the beverage; stabilizer in an amount of 0.1-1% by weight of the beverage, wherein the stabilizer is high methoxy pectin; and a fruit and/or vegetable preparation in an amount of 23.5 to 74.25% by weight of the beverage. | 1,600 |
1,028 | 14,962,805 | 1,648 | The present invention provides a particle comprising a polypeptide and at least one antigen, and a composition comprising thereof. | 1. A particle comprising a virus structural polypeptide and at least one antigen, wherein said virus structural polypeptide comprises at least one first attachment site and said at least one antigen comprises at least one second attachment site, and wherein said virus structural polypeptide and said antigen are linked through said at least one first and said at least one second attachment site, and wherein said particle is virus like particle. 2. The particle according to claim 1, wherein said virus like particle is derived from alphavirus or Flavivirus. 3. The particle according to claim 2, wherein said alphavirus is Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV). 4. The particle according to claim 1, wherein said polypeptide is virus structural polypeptide comprising the capsid and/or the envelope proteins E3, E2, 6K and E1. 5. The particle according to claim 4, wherein said at least one antigen is inserted in to E2 of the envelope protein. 6. The particle according to claim 1, wherein said antigen is at least one selected from the group consisting of self antigens and cancer antigens. 7. The particle according to claim 1, wherein said antigen is a polypeptide derived from TNF-α, CD20 or CTLA4. 8. The particle according claim 1, wherein said polypeptide and said at least one antigen is
i) a polypeptide derived from Chikungunya virus (CHIKV) and a polypeptide of TNF-α; ii) a polypeptide derived from Chikungunya virus (CHIKV) and a polypeptide of CD20; iii) a polypeptide derived from Venezuelan equine encephalitis virus (VEEV) and a polypeptide of TNF-α; or iv) a polypeptide derived from Venezuelan equine encephalitis virus (VEEV) and a polypeptide of CD20 v) a polypeptide derived from Venezuelan equine encephalitis virus (VEEV) and a polypeptide of CTLA4. 9. The particle according to claim 1, wherein said at least one antigen and said polypeptide are expressed as a fusion protein. 10. The particle according to claim 9, wherein said at least one antigen are fused with said polypeptide, wherein one or two linkers intervenes between N-terminal residue of said antigen and said polypeptide and/or between C-terminal residue of said antigen and said polypeptide. 11. The particle according to claim 10, wherein said at least one antigen is inserted between residues 519 and 520 of SEQ ID Nos.1 or 2, between residues 530 and 531 of SEQ ID Nos.1 or 2, between residues 531 and 532 of SEQ ID Nos.1 or 2 or between residues 532 and 533 of SEQ ID Nos.1 or 2. 12. The particle according to claim 9, wherein said fusion protein is a protein consisting of an amino acid sequence represented by SEQ ID Nos. 4, 5, 6, 7 or 8. 13. The particle according to claim 9, wherein said fusion protein is derived from a protein consisting of an amino acid sequence which has a sequence identity of 90% or more with an amino acid sequence represented by SEQ ID Nos. 4, 5, 6, 7 or 8. 14. A composition comprising the particle according to claim 1. 15. A vaccine composition comprising the particle according to claim 1. 16. A method of producing an antibody, comprising administering the particle according to claim 1 to a mammal. 17. The method according to claim 16, wherein said antibody is a monoclonal antibody. 18. A method of immunomodulation, comprising administering an immunologically effective amount of the composition of claim 14 to a mammal. 19. A method of treating an autoimmune disease, comprising administering an immunologically effective amount of the composition of claim 14 to a mammal. 20. A method of treating cancer, comprising administering an effective amount of the composition of claim 14 to a mammal. | The present invention provides a particle comprising a polypeptide and at least one antigen, and a composition comprising thereof.1. A particle comprising a virus structural polypeptide and at least one antigen, wherein said virus structural polypeptide comprises at least one first attachment site and said at least one antigen comprises at least one second attachment site, and wherein said virus structural polypeptide and said antigen are linked through said at least one first and said at least one second attachment site, and wherein said particle is virus like particle. 2. The particle according to claim 1, wherein said virus like particle is derived from alphavirus or Flavivirus. 3. The particle according to claim 2, wherein said alphavirus is Chikungunya virus (CHIKV) or Venezuelan equine encephalitis virus (VEEV). 4. The particle according to claim 1, wherein said polypeptide is virus structural polypeptide comprising the capsid and/or the envelope proteins E3, E2, 6K and E1. 5. The particle according to claim 4, wherein said at least one antigen is inserted in to E2 of the envelope protein. 6. The particle according to claim 1, wherein said antigen is at least one selected from the group consisting of self antigens and cancer antigens. 7. The particle according to claim 1, wherein said antigen is a polypeptide derived from TNF-α, CD20 or CTLA4. 8. The particle according claim 1, wherein said polypeptide and said at least one antigen is
i) a polypeptide derived from Chikungunya virus (CHIKV) and a polypeptide of TNF-α; ii) a polypeptide derived from Chikungunya virus (CHIKV) and a polypeptide of CD20; iii) a polypeptide derived from Venezuelan equine encephalitis virus (VEEV) and a polypeptide of TNF-α; or iv) a polypeptide derived from Venezuelan equine encephalitis virus (VEEV) and a polypeptide of CD20 v) a polypeptide derived from Venezuelan equine encephalitis virus (VEEV) and a polypeptide of CTLA4. 9. The particle according to claim 1, wherein said at least one antigen and said polypeptide are expressed as a fusion protein. 10. The particle according to claim 9, wherein said at least one antigen are fused with said polypeptide, wherein one or two linkers intervenes between N-terminal residue of said antigen and said polypeptide and/or between C-terminal residue of said antigen and said polypeptide. 11. The particle according to claim 10, wherein said at least one antigen is inserted between residues 519 and 520 of SEQ ID Nos.1 or 2, between residues 530 and 531 of SEQ ID Nos.1 or 2, between residues 531 and 532 of SEQ ID Nos.1 or 2 or between residues 532 and 533 of SEQ ID Nos.1 or 2. 12. The particle according to claim 9, wherein said fusion protein is a protein consisting of an amino acid sequence represented by SEQ ID Nos. 4, 5, 6, 7 or 8. 13. The particle according to claim 9, wherein said fusion protein is derived from a protein consisting of an amino acid sequence which has a sequence identity of 90% or more with an amino acid sequence represented by SEQ ID Nos. 4, 5, 6, 7 or 8. 14. A composition comprising the particle according to claim 1. 15. A vaccine composition comprising the particle according to claim 1. 16. A method of producing an antibody, comprising administering the particle according to claim 1 to a mammal. 17. The method according to claim 16, wherein said antibody is a monoclonal antibody. 18. A method of immunomodulation, comprising administering an immunologically effective amount of the composition of claim 14 to a mammal. 19. A method of treating an autoimmune disease, comprising administering an immunologically effective amount of the composition of claim 14 to a mammal. 20. A method of treating cancer, comprising administering an effective amount of the composition of claim 14 to a mammal. | 1,600 |
1,029 | 14,641,009 | 1,619 | Drug delivery devices and methods are provided for administering gemcitabine to a patient in need of treatment of bladder cancer by intravesically administering gemcitabine into the bladder of the patient to achieve a sustained concentration of the gemcitabine in urine in the bladder sufficient to produce a therapeutically effective concentration of the gemcitabine in the tissues of the bladder. In embodiments, the local administration into the patient's bladder is at a mean average amount of from 1 mg/day to about 300 mg/day of the gemcitabine (FBE). | 1. A method of treatment of bladder cancer in a patient, comprising:
locally administering gemcitabine into the urinary bladder of the patient to achieve a sustained concentration of the gemcitabine in urine in the bladder sufficient to produce a therapeutic concentration of the gemcitabine in the bladder tissues, wherein the locally administering into the patient's bladder is at a mean average amount of from 1 mg/day to about 300 mg/day of the gemcitabine (FBE). 2. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of from 1 mg/day to 200 mg/day of the gemcitabine (FBE). 3. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of from 5 mg/day to 100 mg/day of the gemcitabine (FBE). 4. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of from 10 mg/day to 50 mg/day of the gemcitabine (FBE). 5. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of from 15 mg/day to 25 mg/day of the gemcitabine (FBE). 6. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of about 20 mg/day of the gemcitabine (FBE). 7. The method of claim 1, wherein the locally administering into the patient's bladder is continuous over a period from 1 day to 30 days. 8. The method of claim 1, wherein the locally administering into the patient's bladder is intermittent over a period from 1 day to 30 days. 9. The method of claim 1, wherein the locally administering into the patient's bladder is intermittent or continuous over a period from 3 days to 21 days. 10. The method of claim 1, wherein the locally administering into the patient's bladder is continuous over a period from 7 day to 21 days. 11. The method of claim 1, wherein the gemcitabine is delivered into the bladder from an intravesical drug delivery device which continuously releases the gemcitabine into the urine in the bladder over a sustained period. 12. The method of claim 11, wherein the sustained period is 1 day to 21 days. 13. The method of claim 11, wherein the intravesical drug delivery device comprises a housing which contains and controllably releases the gemcitabine and is elastically deformable between a retention shape configured to retain the device in a patient's bladder and a deployment shape for passage of the device through the patient's urethra. 14. The method of claim 13, wherein the gemcitabine contained in the housing is in a non-liquid form. 15. The method of claim 14, wherein the non-liquid form is selected from the group consisting of tablets, granules, semisolids, capsules, and combinations thereof. 16. The method of claim 1, wherein the gemcitabine is delivered into the bladder from a coating substance applied to the bladder, which coating substance releases the gemcitabine into the urine in the bladder over a sustained period. 17. The method of claim 16, wherein the coating substance comprises a mucoadhesive formulation. 18. The method of claim 17, wherein the sustained period is from 1 day to 14 days. 19. The method of claim 17, wherein the sustained period is from 1 day to 7 days. 20. The method of claim 1, wherein the locally administering comprises pumping a liquid form of the gemcitabine into the bladder through a urethral or suprapubic catheter which is deployed into the bladder. 21. The method of claim 20, wherein the locally administering into the patient's bladder is continuous or intermittent over a period from 1 day to 7 days. 22. The method of claim 1, further comprising administering at least a second therapeutic agent to the patient. 23. The method of claim 22, wherein the second therapeutic agent is administered intravesically. 24. The method of claim 1, further comprising administering urea or another solubility altering agent into the bladder in an amount effective to enhance or otherwise alter solubilization of the gemcitabine. 25. The method of claim 24, wherein the urea or other solubility altering agent is released from an intravesical device releasing the gemcitabine. 26. A drug delivery device comprising:
a housing configured for intravesical insertion; and a dosage form comprising gemcitabine, wherein the housing holds the dosage form and is configured to release the gemcitabine into the bladder in an amount therapeutically effective for the treatment of the bladder, wherein the device is configured to release gemcitabine into the bladder at a mean average amount of from 1 mg/day to about 300 mg/day of the gemcitabine. 27. The device of claim 26, wherein the housing releases the gemcitabine by diffusion through a drug permeable polymeric wall. 28. The device of claim 26, wherein the housing releases the gemcitabine without a predefined release aperture. 29. The device of claim 26, wherein the housing comprises a release orifice in communication with a drug reservoir in which the gemcitabine is contained along with (i) a viscosity enhancing agent, (ii) an osmotic agent, or (iii) a combination of a viscosity enhancing agent and an osmotic agent. 30. The device of claim 29, wherein the gemcitabine is provided in a first region comprising one or more tablets and the osmotic agent and/or viscosity enhancing agent is/are provided in a second region comprising one or more tablets, wherein the first and second regions are discrete spaces within the drug reservoir. | Drug delivery devices and methods are provided for administering gemcitabine to a patient in need of treatment of bladder cancer by intravesically administering gemcitabine into the bladder of the patient to achieve a sustained concentration of the gemcitabine in urine in the bladder sufficient to produce a therapeutically effective concentration of the gemcitabine in the tissues of the bladder. In embodiments, the local administration into the patient's bladder is at a mean average amount of from 1 mg/day to about 300 mg/day of the gemcitabine (FBE).1. A method of treatment of bladder cancer in a patient, comprising:
locally administering gemcitabine into the urinary bladder of the patient to achieve a sustained concentration of the gemcitabine in urine in the bladder sufficient to produce a therapeutic concentration of the gemcitabine in the bladder tissues, wherein the locally administering into the patient's bladder is at a mean average amount of from 1 mg/day to about 300 mg/day of the gemcitabine (FBE). 2. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of from 1 mg/day to 200 mg/day of the gemcitabine (FBE). 3. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of from 5 mg/day to 100 mg/day of the gemcitabine (FBE). 4. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of from 10 mg/day to 50 mg/day of the gemcitabine (FBE). 5. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of from 15 mg/day to 25 mg/day of the gemcitabine (FBE). 6. The method of claim 1, wherein the locally administering into the patient's bladder is at a mean average amount of about 20 mg/day of the gemcitabine (FBE). 7. The method of claim 1, wherein the locally administering into the patient's bladder is continuous over a period from 1 day to 30 days. 8. The method of claim 1, wherein the locally administering into the patient's bladder is intermittent over a period from 1 day to 30 days. 9. The method of claim 1, wherein the locally administering into the patient's bladder is intermittent or continuous over a period from 3 days to 21 days. 10. The method of claim 1, wherein the locally administering into the patient's bladder is continuous over a period from 7 day to 21 days. 11. The method of claim 1, wherein the gemcitabine is delivered into the bladder from an intravesical drug delivery device which continuously releases the gemcitabine into the urine in the bladder over a sustained period. 12. The method of claim 11, wherein the sustained period is 1 day to 21 days. 13. The method of claim 11, wherein the intravesical drug delivery device comprises a housing which contains and controllably releases the gemcitabine and is elastically deformable between a retention shape configured to retain the device in a patient's bladder and a deployment shape for passage of the device through the patient's urethra. 14. The method of claim 13, wherein the gemcitabine contained in the housing is in a non-liquid form. 15. The method of claim 14, wherein the non-liquid form is selected from the group consisting of tablets, granules, semisolids, capsules, and combinations thereof. 16. The method of claim 1, wherein the gemcitabine is delivered into the bladder from a coating substance applied to the bladder, which coating substance releases the gemcitabine into the urine in the bladder over a sustained period. 17. The method of claim 16, wherein the coating substance comprises a mucoadhesive formulation. 18. The method of claim 17, wherein the sustained period is from 1 day to 14 days. 19. The method of claim 17, wherein the sustained period is from 1 day to 7 days. 20. The method of claim 1, wherein the locally administering comprises pumping a liquid form of the gemcitabine into the bladder through a urethral or suprapubic catheter which is deployed into the bladder. 21. The method of claim 20, wherein the locally administering into the patient's bladder is continuous or intermittent over a period from 1 day to 7 days. 22. The method of claim 1, further comprising administering at least a second therapeutic agent to the patient. 23. The method of claim 22, wherein the second therapeutic agent is administered intravesically. 24. The method of claim 1, further comprising administering urea or another solubility altering agent into the bladder in an amount effective to enhance or otherwise alter solubilization of the gemcitabine. 25. The method of claim 24, wherein the urea or other solubility altering agent is released from an intravesical device releasing the gemcitabine. 26. A drug delivery device comprising:
a housing configured for intravesical insertion; and a dosage form comprising gemcitabine, wherein the housing holds the dosage form and is configured to release the gemcitabine into the bladder in an amount therapeutically effective for the treatment of the bladder, wherein the device is configured to release gemcitabine into the bladder at a mean average amount of from 1 mg/day to about 300 mg/day of the gemcitabine. 27. The device of claim 26, wherein the housing releases the gemcitabine by diffusion through a drug permeable polymeric wall. 28. The device of claim 26, wherein the housing releases the gemcitabine without a predefined release aperture. 29. The device of claim 26, wherein the housing comprises a release orifice in communication with a drug reservoir in which the gemcitabine is contained along with (i) a viscosity enhancing agent, (ii) an osmotic agent, or (iii) a combination of a viscosity enhancing agent and an osmotic agent. 30. The device of claim 29, wherein the gemcitabine is provided in a first region comprising one or more tablets and the osmotic agent and/or viscosity enhancing agent is/are provided in a second region comprising one or more tablets, wherein the first and second regions are discrete spaces within the drug reservoir. | 1,600 |
1,030 | 15,746,424 | 1,699 | The abstract particularly describes the nature of this invention and the manner in which it is to be performed:—
1. Specific process and apparatus developed
The inventor has prepared a novel process to speedily and accurately detect pathogens present in a fluid sample, identify the pathogens and it's count and determine the level of their susceptibility levels to multiple antimicrobial agents and portability of the apparatus for speedy determination of antibiotic susceptibility levels of pre-separated pathogens from a biological sample cultured in a multi-well strip, in any field location
2. Disadvantage and shortcomings in earlier method and benefit of this invention
It provides result in a ready to use format in four hours time from the start of the assay as against the waiting period of 48 to 72 hours for a sample cultured and tested in a lab using the conventional clinical microbiology. | 1-10. (canceled) 11. A process for rapidly detecting bacteria in a urine sample along with corresponding susceptibility levels to multiple antibiotics, the process comprising of the following steps:
a) collecting the urine sample in a sterile container; b) preparing a bacterial growth media aliquot, wherein the bacterial growth media contains chromogenic indicators of bacterial growth; c) aspirating the urine sample through a filter into a syringe; d) separating the filter from the syringe, whereby the filter has the bacteria from the urine sample on it; e) filling a syringe with the bacterial growth media aliquot; f) attaching the filter with bacteria to the syringe with the bacterial growth media aliquot; g) pushing the bacterial growth media through the filter into a container, whereby the bacteria are suspended in the bacterial growth media collected in the container; h) dispensing the suspended bacteria into a plurality of wells in a strip, at least two wells in the plurality of wells having two different antibiotics, and at least another reference well having no antibiotics; i) incubating the wells for not less than 3 hours at a temperature between approximately 30° C. to 40° C.; and e) detecting, following said incubation, bacterial growth by measuring intensity of a color and turbidity in each of the wells relative to that in the reference well and a specific provided reference. 12. The process of claim 11 wherein the specific provided reference provides specific chromogenic endpoints to identify one or more selected from the group consisting of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterococcus sp. 13. The process according to claim 12, wherein comparison of bacterial growth to the specific chromogenic endpoints allows estimation of a bacterial load and/or a number of bacterial species present in the urine sample. 14. The process of claim 11, wherein the process is implemented using a portable apparatus having dedicated software to provide results on identification and antibiotic susceptibility of pathogens in the urine sample to a person able to handle prepackaged consumables, wherein the prepackaged consumables include a growth media in a vial, sterile water, a syringe, multi-wells, and a filter. 15. The prepackaged consumables of claim 14 further including a reference identification strip having the specific chromogenic endpoints. 16. The process of claim 11, wherein the antibiotics screened are selected from the group consisting of amoxicillin, gentamicin, amikacin, cefepime, ofloxacin, ciprofloxacin, ceftriaxone, piperacillin, cefatoxamine, kanamycin, cefuroxime, tobramycin, levofloxacin, and ampicillin. 17. The process of claim 11 that is implemented at a medical facility or a home of a patient. 18. A kit for detection and separation of microbiological pathogens present in a urine or ascites sample comprising:
at least one container that is used for collecting the urine or ascites sample; at least one vial containing culture growth media; one or more multi-well strips wherein at least one well is pre-loaded with an antibiotic and at least one well is a reference well, wherein the reference well does not contain any antibiotic; at least one vial containing sterile water; and
a needle,
a syringe;
a cap; and
at least one filter for filtering out bacteria from the clinical or biological sample. 19. The kit of claim 18 further comprising a specific provided reference having specific chromogenic endpoints to identify and quantify bacteria. 20. An apparatus having a pre-programmed microprocessor for rapid determination of antibiotic sensitivity profile of bacteria in a urine sample, the pre-programmed microprocessor including code for:
a) identification of bacteria cultured inside a multi-well strip, which includes wells preloaded with a antibiotics and at least one reference well that lacks antibiotics; b) determination of the bacterial load; and c) determining the antibiotic susceptibility level of the cultured bacteria. 21. The apparatus according to claim 20 wherein the microprocessor is pre-programmed to calculate the susceptibility level of the pre-separated pathogens grown in the corresponding well on the basis of a data synchronization, which includes acquiring data from a well of the multi-well strip; and comparing it to the reference well and provided expected values for particular bacterial loads and types. | The abstract particularly describes the nature of this invention and the manner in which it is to be performed:—
1. Specific process and apparatus developed
The inventor has prepared a novel process to speedily and accurately detect pathogens present in a fluid sample, identify the pathogens and it's count and determine the level of their susceptibility levels to multiple antimicrobial agents and portability of the apparatus for speedy determination of antibiotic susceptibility levels of pre-separated pathogens from a biological sample cultured in a multi-well strip, in any field location
2. Disadvantage and shortcomings in earlier method and benefit of this invention
It provides result in a ready to use format in four hours time from the start of the assay as against the waiting period of 48 to 72 hours for a sample cultured and tested in a lab using the conventional clinical microbiology.1-10. (canceled) 11. A process for rapidly detecting bacteria in a urine sample along with corresponding susceptibility levels to multiple antibiotics, the process comprising of the following steps:
a) collecting the urine sample in a sterile container; b) preparing a bacterial growth media aliquot, wherein the bacterial growth media contains chromogenic indicators of bacterial growth; c) aspirating the urine sample through a filter into a syringe; d) separating the filter from the syringe, whereby the filter has the bacteria from the urine sample on it; e) filling a syringe with the bacterial growth media aliquot; f) attaching the filter with bacteria to the syringe with the bacterial growth media aliquot; g) pushing the bacterial growth media through the filter into a container, whereby the bacteria are suspended in the bacterial growth media collected in the container; h) dispensing the suspended bacteria into a plurality of wells in a strip, at least two wells in the plurality of wells having two different antibiotics, and at least another reference well having no antibiotics; i) incubating the wells for not less than 3 hours at a temperature between approximately 30° C. to 40° C.; and e) detecting, following said incubation, bacterial growth by measuring intensity of a color and turbidity in each of the wells relative to that in the reference well and a specific provided reference. 12. The process of claim 11 wherein the specific provided reference provides specific chromogenic endpoints to identify one or more selected from the group consisting of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterococcus sp. 13. The process according to claim 12, wherein comparison of bacterial growth to the specific chromogenic endpoints allows estimation of a bacterial load and/or a number of bacterial species present in the urine sample. 14. The process of claim 11, wherein the process is implemented using a portable apparatus having dedicated software to provide results on identification and antibiotic susceptibility of pathogens in the urine sample to a person able to handle prepackaged consumables, wherein the prepackaged consumables include a growth media in a vial, sterile water, a syringe, multi-wells, and a filter. 15. The prepackaged consumables of claim 14 further including a reference identification strip having the specific chromogenic endpoints. 16. The process of claim 11, wherein the antibiotics screened are selected from the group consisting of amoxicillin, gentamicin, amikacin, cefepime, ofloxacin, ciprofloxacin, ceftriaxone, piperacillin, cefatoxamine, kanamycin, cefuroxime, tobramycin, levofloxacin, and ampicillin. 17. The process of claim 11 that is implemented at a medical facility or a home of a patient. 18. A kit for detection and separation of microbiological pathogens present in a urine or ascites sample comprising:
at least one container that is used for collecting the urine or ascites sample; at least one vial containing culture growth media; one or more multi-well strips wherein at least one well is pre-loaded with an antibiotic and at least one well is a reference well, wherein the reference well does not contain any antibiotic; at least one vial containing sterile water; and
a needle,
a syringe;
a cap; and
at least one filter for filtering out bacteria from the clinical or biological sample. 19. The kit of claim 18 further comprising a specific provided reference having specific chromogenic endpoints to identify and quantify bacteria. 20. An apparatus having a pre-programmed microprocessor for rapid determination of antibiotic sensitivity profile of bacteria in a urine sample, the pre-programmed microprocessor including code for:
a) identification of bacteria cultured inside a multi-well strip, which includes wells preloaded with a antibiotics and at least one reference well that lacks antibiotics; b) determination of the bacterial load; and c) determining the antibiotic susceptibility level of the cultured bacteria. 21. The apparatus according to claim 20 wherein the microprocessor is pre-programmed to calculate the susceptibility level of the pre-separated pathogens grown in the corresponding well on the basis of a data synchronization, which includes acquiring data from a well of the multi-well strip; and comparing it to the reference well and provided expected values for particular bacterial loads and types. | 1,600 |
1,031 | 12,844,440 | 1,639 | We describe apparatuses, method, reagents, and kits for conducting assays as well as process for their preparation. They are particularly well suited for conducting automated sampling, sample preparation, and analysis in a multi-well plate assay format. For example, they may be used for automated analysis of liquid samples in a clinical point of care setting. | 1. A kit for conducting luminescence assays in multi-well plates, the kit comprising:
(a) a multi-well assay test plate comprising a plurality of assay wells for said assay; (b) an auxiliary plate comprising a plurality of auxiliary wells, said auxiliary well comprising dry assay reagents for use in said assay with said assay test plate. 2. A kit according to claim 1 wherein a well of said test plate comprises a plurality of distinct assay domains, at least two of said domains comprising reagents for measuring different analytes. 3. A kit according to any one of the preceding claims wherein said auxiliary plate comprises an identifier comprising assay information used to identify an element selected from the group consisting of (i) said auxiliary plate, (ii) one or more auxiliary wells within said auxiliary plate, (iii) a reagent and/or sample that has been or will be used with said auxiliary plate, (iv) said test plate, (v) one or more wells within said test plate, (vi) a reagent and/or sample that has been or will be used with said test plate, and (vii) combinations thereof. 4. A kit according to claim 3 wherein said identifier comprises test plate information identifying a test plate for use with said auxiliary plate. 5. A kit according to claim 4 wherein said test plate information comprises test plate lot information. 6. A kit according to claim 4 wherein said test plate information comprises a test plate identification number. 7. A kit according to claim 1 wherein said assay test plate comprises an identifier comprising assay information used to identify an element selected from the group consisting of (i) said auxiliary plate, (ii) one or more auxiliary wells within said auxiliary plate, (iii) a reagent and/or sample that has been or will be used with said auxiliary plate, (iv) said test plate, (v) one or more wells within said test plate, (vi) a reagent and/or sample that has been or will be used with said test plate, and (vii) combinations thereof. 8. A kit according to claim 7 wherein said identifier comprises auxiliary plate information identifying an auxiliary plate for use with said test plate. 9. A kit according to claim 8 wherein said auxiliary plate information comprises auxiliary plate lot information. 10. A kit according to claim 8 wherein said auxiliary plate information comprises an auxiliary plate identification number. 11. A kit according to claim 1 wherein said plurality of auxiliary wells is a multiple of the number of assay wells in said assay test plate. 12. A kit according to claim 1 wherein said auxiliary plate comprises twice as many auxiliary wells as assay wells in said assay test plate. 13. A kit according to claim 1 wherein said auxiliary plate comprises four times as many auxiliary wells as assay wells in said assay test plate. 14. A kit according to claim 1 wherein said assay test plate further comprises a plurality of elements selected from the group consisting of a plate top, plate bottom, working electrodes, counter electrodes, reference electrodes, dielectric materials, electrical connections, dried or liquid assay reagents, and combinations thereof. 15. A kit according to claim 1 wherein said auxiliary plate further comprises a set of auxiliary wells, said set comprising adjacent auxiliary wells, wherein said set of auxiliary wells comprises reagents for an assay in a well of said assay test plate. 16. A kit according to claim 15 wherein said set comprises four adjacent auxiliary wells. 17. A kit according to claim 16 wherein said four adjacent auxiliary wells are arranged in a square. 18. A kit according to claim 16 wherein said four adjacent auxiliary wells are arranged in a row. 19. A kit according to claim 15 wherein a auxiliary well of said set of auxiliary wells is a dilution well. 20. A kit according to claim 15 wherein a auxiliary well of said set of auxiliary wells comprises pre-treated beads. 21. A kit according to claim 20 wherein said pre-treated beads are magnetic. 22. A kit according to claim 21 wherein said pre-treated beads comprise a coating selected from the group consisting of streptavidin, biotin, and avidin. 23. A kit according to claim 22 wherein one or more reagents in said set of auxiliary wells comprise a binding partner of said coating. 24. A kit according to claim 15 wherein at least one auxiliary well of said set comprises desiccant and the auxiliary plate comprises a seal. 25. A kit according to claim 24 wherein said at least one auxiliary well of said set is connected to an additional auxiliary well of said set via an air passage. 26. A kit according to claim 24 wherein said at least one auxiliary well of said set is connected to all auxiliary wells of said set via said air passage. 27. A kit according to claim 3 wherein said information is consumable information selected from the group consisting of lot identification information; lot specific analysis parameters, manufacturing process information, raw materials information, expiration date; calibration data; threshold information; the location of individual assay reagents and/or samples within one or more wells and/or auxiliary wells of said auxiliary plate and/or said test plate; Material Safety Data Sheet (MSDS) information, and combinations thereof. 28. A kit according to claim 3 wherein said information is sample information selected from the group consisting of the intended location of samples within said one or more wells of the test plate; assay results obtained on said test plate for said sample; identity of samples that have been and/or will be assayed in said test plate; and combinations thereof. 29. A kit according to claim 3 wherein said information is chain of custody information. 30. A kit according to claim 29 wherein said chain of custody information includes information regarding the control, transfer and/or analysis of a sample. 31. A kit according to claim 29 wherein said chain of custody information includes information regarding the control, transfer and/or analysis of a reagent. 32. A kit according to claim 29 wherein said information is chain of custody information regarding the control, transfer and/or manufacture of said auxiliary plate and/or test plate. 33. A kit according to claim 29 wherein said chain of custody information is selected from the group consisting of user identification; time and date stamp for said assay; location of an assay system using said auxiliary plate and test plate during said assay; calibration and QC status of said assay system during said assay, QC status of said auxiliary plate; QC status of said test plate, custody and/or location information for said auxiliary plate and/or test plate before and after the conduct of said assay; assay results for said sample; and combinations thereof. 34. A kit according to claim 29 wherein said information is chain of custody information selected from the group consisting of time, date, manufacturing personnel or processing parameters for one or more steps during the manufacture of said auxiliary plate and/or test plate; custody, location and or storage conditions for said auxiliary plate and/or test plate following manufacture and/or betweens steps during the manufacture of said auxiliary plate and/or test plate; and combinations thereof. 35. A kit according to claim 3 wherein said information is auxiliary plate and/or test plate information selected from the group consisting of plate type and structure; location and identity of assay reagents included with said auxiliary plate; location and identify of assay reagents included with said test plate; and combinations thereof. 36. A kit according to claim 3 wherein said assay is a multi-step assay and said information is assay process information that relates to a step or steps of said multi-step assay. 37. A kit according to claim 3 wherein said information is consumable security information selected from the group consisting of information concerning test plate and/or auxiliary plate authentication; information concerning defects in said test plate, auxiliary plate and/or a test site thereof; and combinations thereof. 38. A kit according to claim 1 wherein said test plate comprises capture antibodies for an agent selected from the group consisting of influenza-type A, influenza-type B, RSV, parainfluenza, and adenovirus; and said auxiliary plate comprises detection antibodies for said agent. 39. A kit according to claim 38 wherein said auxiliary plate further comprises desiccant. 40. A kit according to claim 1 wherein said test plate comprises capture antibodies for an agent selected from the group consisting of influenza-type A, influenza-type B, RSV, parainfluenza, adenovirus, influenza-type A (H1), influenza-type A (H2), influenza-type A (H3), influenza-type A (H5), influenza-type A (H7), influenza-type A (H9); and said auxiliary plate comprises one or more reagents selected from the group consisting of HA acidification buffer, HA neutralization buffer, a detection antibody for said agent, NP, and desiccant. 41. A kit according to claim 1 wherein said test plate comprises capture antibodies to a serum biomarker and said auxiliary plate comprises detection antibodies for said biomarker. 42. A kit according to claim 41 wherein said auxiliary plate further comprises desiccant. 43. An auxiliary plate comprising a plurality of assay auxiliary well, said auxiliary well comprising dry assay reagents for use in an assay with a corresponding assay test plate. 44. An auxiliary plate according to claim 43 wherein said auxiliary plate comprises an identifier comprising assay information used to identify an element selected from the group consisting of (i) said auxiliary plate, (ii) one or more auxiliary wells within said auxiliary plate, (iii) a reagent and/or sample that has been or will be used with said auxiliary plate, (iv) said test plate, (v) one or more wells within said test plate, (vi) a reagent and/or sample that has been or will be used with said test plate, and (vii) combinations thereof. 45. An auxiliary plate according to claim 44 wherein said identifier comprises test plate information identifying a test plate for use with said auxiliary plate. 46. An auxiliary plate according to claim 45 wherein said test plate information comprises test plate lot information. 47. An auxiliary plate according to claim 45 wherein said test plate information comprises a test plate identification number. 48. An auxiliary plate according to claim 43 wherein said plurality of auxiliary wells is a multiple of the number of wells in said assay test plate. 49. An auxiliary plate according to claim 48 wherein said auxiliary plate comprises twice as many auxiliary wells as wells in said assay test plate. 50. An auxiliary plate according to claim 48 wherein said auxiliary plate comprises four times as many auxiliary wells as wells in said assay test plate. 51. An auxiliary plate according to claim 43 wherein said auxiliary plate further comprises a set of auxiliary wells, said set comprising adjacent auxiliary wells, wherein said set of auxiliary wells comprises reagents for an assay in a well of said assay test plate. 52. An auxiliary plate according to claim 51 wherein said set comprises four adjacent auxiliary wells. 53. An auxiliary plate according to claim 52 wherein said four adjacent auxiliary wells are arranged in a square. 54. An auxiliary plate according to claim 52 wherein said four adjacent auxiliary wells are arranged in a row. 55. An auxiliary plate according to claim 51 wherein a auxiliary well of said set of auxiliary wells is a dilution well. 56. An auxiliary plate according to claim 51 wherein a auxiliary well of said set of auxiliary wells comprises pre-treated beads. 57. An auxiliary plate according to claim 56 wherein said pre-treated beads are magnetic. 58. An auxiliary plate according to claim 56 wherein said pre-treated beads comprise a coating selected from the group consisting of streptavidin, biotin, and avidin. 59. An auxiliary plate according to claim 58 wherein one or more reagents in said set of auxiliary wells comprise a binding partner of said coating. 60. An auxiliary plate according to claim 51 wherein at least one auxiliary well of said set comprises desiccant and said auxiliary plate comprises a seal. 61. An auxiliary plate according to claim 60 wherein said at least one auxiliary well of said set is connected to an additional auxiliary well of said set via an air passage. 62. An auxiliary plate according to claim 60 wherein said at least one auxiliary well of said set is connected to all auxiliary wells of said set via said air passage. 63. An auxiliary plate according to claim 43 wherein said information is consumable information selected from the group consisting of lot identification information; lot specific analysis parameters, manufacturing process information, raw materials information, expiration date; calibration data; threshold information; the location of individual assay reagents and/or samples within one or more wells and/or auxiliary wells of said auxiliary plate and/or said test plate; Material Safety Data Sheet (MSDS) information, and combinations thereof. 64. An auxiliary plate according to claim 43 wherein said information is sample information selected from the group consisting of the intended location of samples within said one or more auxiliary wells of the auxiliary plate; assay results obtained on said test plate for said sample; identity of samples that have been and/or will be assayed in said test plate; and combinations thereof. 65. An auxiliary plate according to claim 43 wherein said information is chain of custody information. 66. An auxiliary plate according to claim 65 wherein said chain of custody information includes information regarding the control, transfer and/or analysis of a reagent. 67. An auxiliary plate according to claim 65 wherein said information is chain of custody information regarding the control, transfer and/or manufacture of said auxiliary plate and/or test plate. 68. An auxiliary plate according to claim 65 wherein said chain of custody information is selected from the group consisting of user identification; time and date stamp for said assay; location of an assay system using said auxiliary plate and test plate during said assay; calibration and QC status of said assay system during said assay, QC status of said auxiliary plate; QC status of said test plate, custody and/or location information for said auxiliary plate and/or test plate before and after the conduct of said assay; assay results for said sample; and combinations thereof. 69. An auxiliary plate according to claim 65 wherein said information is chain of custody information selected from the group consisting of time, date, manufacturing personnel or processing parameters for one or more steps during the manufacture of said auxiliary plate and/or test plate; custody, location and or storage conditions for said auxiliary plate and/or test plate following manufacture and/or betweens steps during the manufacture of said auxiliary plate and/or test plate; and combinations thereof. 70. An auxiliary plate according to claim 43 wherein said information is auxiliary plate and/or test plate information selected from the group consisting of plate type and structure;
location and identity of assay reagents included with said auxiliary plate; location and identify of assay reagents included with said test plate; and combinations thereof. 71. An auxiliary plate according to claim 43 wherein said assay is a multi-step assay and said information is assay process information that relates to a step or step(s) of said multi-step assay. 72. An auxiliary plate according to claim 43 wherein said information is consumable security information selected from the group consisting of information concerning test plate and/or auxiliary plate authentication; information concerning defects in said test plate, auxiliary plate and/or a test well thereof; and combinations thereof. 73. An auxiliary plate according to claim 43 wherein said auxiliary plate comprises detection antibodies for an agent selected from the group consisting of influenza-type A, influenza-type B, RSV, Parainfluenza, and adenovirus. 74. An auxiliary plate according to claim 73 wherein said auxiliary plate further comprises desiccant. 75. An auxiliary plate according to claim 43 wherein said auxiliary plate comprises one or more reagents selected from the group consisting of HA acidification buffer, HA neutralization buffer, NP, desiccant, and a detection antibody for an agent selected from the group consisting of influenza-type A, influenza-type B, RSV, parainfluenza, adenovirus, influenza-type A (H1), influenza-type A (H2), influenza-type A (H3), influenza-type A (H5), influenza-type A (H7), influenza-type A (H9). 76. An auxiliary plate according to claim 43 wherein said auxiliary plate comprises detection antibodies to a serum biomarker. 77. An auxiliary plate according to claim 76 wherein said auxiliary plate further comprises desiccant. 78. An apparatus for conducting a measurement in a multi-well assay test plate, said apparatus comprising
(a) a subassembly capable of supporting and translating said test plate to one or more components of said apparatus; and (b) an auxiliary plate subassembly, wherein said auxiliary plate comprises a plurality of auxiliary wells comprising dry assay reagents for use in an assay with said test plate. 79. An apparatus according to claim 78 further comprising a pipettor subassembly that delivers sample and/or reagent to and from a well of said test plate and/or an auxiliary well of said auxiliary plate. 80. An apparatus according to claim 79 wherein said pipettor subassembly comprises a component selected from the group consisting of a pump, a plate piercing probe, a pipetting probe and an ultrasonic sensor. 81. An apparatus according to claim 78 wherein said subassembly capable of supporting and translating said test plate comprises a plate introduction aperture and a plate translation stage. 82. An apparatus according to claim 81 further comprising a plate stacker adjacent to said plate introduction aperture. 83. An apparatus according to claim 82 further comprising a plate elevator comprising a plate lifting platform that can be raised and lowered onto said plate translation stage. 84. An apparatus of claim 81 further comprising (i) an input plate introduction aperture comprising an input plate stacker and (ii) an output plate introduction aperture comprising an output plate stacker. 85. An apparatus of any one of claims 81 or 84 wherein said plate stacker can accommodate more than one test plate. 86. An apparatus of claim 81 wherein said subassembly is a light-tight enclosure and said plate introduction aperture comprises a sliding light-tight door. 87. An apparatus of claim 81 wherein said subassembly further comprises a component selected from the group consisting of a thermoelectric heater/cooler, a desiccant chamber, and an identifier controller. 88. An apparatus of claim 86 further comprising an imaging system mounted to an imaging aperture in the light-tight enclosure. 89. An apparatus of claim 81 wherein said plate translation stage is configured to position a well of a test plate in proximity to one or more components of said subassembly selected from the group consisting of said plate elevator, a well-wash subassembly, and an imaging system. 90. An apparatus of claim 89 wherein said well-wash subassembly comprises a seal removal tool, a well-wash head, a wash station, and fluidic connectors to a liquid reagent subassembly. 91. An apparatus of claim 90 wherein said well-wash head comprises a pipetting probe and a pipetting translation stage for translating said pipetting probe in a vertical direction. 92. An apparatus of claim 91 wherein said pipetting probe comprises a dispensing tube and a plurality of aspiration tubes. 93. An apparatus of claim 81 wherein said auxiliary plate subassembly comprises an auxiliary plate introduction aperture and a plate support. 94. An apparatus of claim 93 wherein said auxiliary plate subassembly further comprises a housing comprising two or more compartments, each compartment comprising said auxiliary plate introduction aperture and said plate support. 95. An apparatus of claim 94 wherein said compartments comprise a component selected from the group consisting of an identifier controller, a thermoelectric heater/cooler, and a desiccant chamber. 96. A method for conducting a measurement in a multi-well assay test plate, said method comprising the steps of:
(a) dispensing sample and/or reagent into an auxiliary well of an auxiliary plate, said auxiliary plate comprising a plurality of auxiliary wells, said plurality of auxiliary wells comprising dry assay reagents for use in an assay with said test plate; and (b) transferring sample and/or reagent from said auxiliary well to a well of a said assay test plate. 97. A method according to claim 96 wherein said dispensing step (a) comprises pre-treating said sample and/or reagent in said auxiliary well. 98. A method according to claim 97 wherein said transferring step (b) comprises dispensing pre-treated sample and/or reagent from said auxiliary well to said well of said test plate. 99. A method according to claim 96 wherein said assay test plate is supported on a plate translation stage and said method comprises translating said test plate via said plate translation stage to one or more components of said apparatus. 100. A method according to claim 99 wherein said method further comprises placing said assay test plate through a plate introduction aperture onto a plate stacker adjacent said plate translation stage. 101. A method according to claim 100 wherein said method further comprises lowering said assay test plate from said plate stacker to said plate translation stage. 102. A method according to claim 101 wherein said method further comprises (i) placing said assay test plate through an input plate introduction aperture comprising an input plate stacker, (ii) lowering said assay test plate from said input plate stacker to said plate translation stage, (iii) translating said assay test plate to one or more components of said apparatus to conduct said measurement, (iv) raising said assay test plate from said plate translation stage to an output plate stacker, and (v) removing said assay test plate from an output plate introduction aperture. 103. A method according to claim 96 wherein said method further comprises repeating steps (a) and (b) in an additional auxiliary well of said auxiliary plate and an additional test well of said test plate. 104. A method for conducting a measurement in a multi-well assay test plate, said method comprising the steps of:
(a) dispensing sample and/or reagent into an auxiliary well of a set of auxiliary wells in an auxiliary plate, said auxiliary plate comprising a plurality of auxiliary wells, said plurality of auxiliary wells comprising dry assay reagents for use in an assay with said test plate; (b) pre-treating said sample and/or reagent in one or more auxiliary wells of said set; (c) dispensing pre-treated sample and/or reagent from said one or more auxiliary wells of said set to a well of a said assay test plate; (d) repeating steps (a)-(c) with an additional sample and reagents in an additional set of auxiliary wells in said auxiliary plate, and an additional well of said test plate; (e) ejecting a used auxiliary plate; (f) repeating steps (a)-(c) with an additional auxiliary plate; and (g) ejecting a used assay test plate. 105. A well-wash subassembly comprising a multi-tube array comprising a central dispensing tube element surrounded by a plurality of aspiration tube elements. 106. A well-wash subassembly according to claim 105 wherein said multi-tube array comprises at least two dispensing tube elements at the center of the array. 107. A well-wash subassembly according to claim 106 wherein said dispensing tube elements comprise an independent fluid channel for buffers and/or diluents used during an assay. 108. A well-wash subassembly according to claim 105 wherein said aspiration tube elements surround the dispensing tube elements and said aspiration tube elements are positioned to align with the outer portions of a well bottom of a multi-well test plate. 109. A well-wash subassembly according to claim 105 wherein said aspiration tube elements are independently connected to dedicated fluidic lines. 110. A well-wash subassembly according to claim 105 wherein said multi-tube array comprises at least four aspiration tube elements. 111. An apparatus according to claim 80 wherein said pipettor subassembly comprises a reflective sensor comprising an infra-red LED and a phototransistor, wherein said LED and phototransistor are positioned in said subassembly to detect the presence or absence of a pipetting tip in said subassembly. | We describe apparatuses, method, reagents, and kits for conducting assays as well as process for their preparation. They are particularly well suited for conducting automated sampling, sample preparation, and analysis in a multi-well plate assay format. For example, they may be used for automated analysis of liquid samples in a clinical point of care setting.1. A kit for conducting luminescence assays in multi-well plates, the kit comprising:
(a) a multi-well assay test plate comprising a plurality of assay wells for said assay; (b) an auxiliary plate comprising a plurality of auxiliary wells, said auxiliary well comprising dry assay reagents for use in said assay with said assay test plate. 2. A kit according to claim 1 wherein a well of said test plate comprises a plurality of distinct assay domains, at least two of said domains comprising reagents for measuring different analytes. 3. A kit according to any one of the preceding claims wherein said auxiliary plate comprises an identifier comprising assay information used to identify an element selected from the group consisting of (i) said auxiliary plate, (ii) one or more auxiliary wells within said auxiliary plate, (iii) a reagent and/or sample that has been or will be used with said auxiliary plate, (iv) said test plate, (v) one or more wells within said test plate, (vi) a reagent and/or sample that has been or will be used with said test plate, and (vii) combinations thereof. 4. A kit according to claim 3 wherein said identifier comprises test plate information identifying a test plate for use with said auxiliary plate. 5. A kit according to claim 4 wherein said test plate information comprises test plate lot information. 6. A kit according to claim 4 wherein said test plate information comprises a test plate identification number. 7. A kit according to claim 1 wherein said assay test plate comprises an identifier comprising assay information used to identify an element selected from the group consisting of (i) said auxiliary plate, (ii) one or more auxiliary wells within said auxiliary plate, (iii) a reagent and/or sample that has been or will be used with said auxiliary plate, (iv) said test plate, (v) one or more wells within said test plate, (vi) a reagent and/or sample that has been or will be used with said test plate, and (vii) combinations thereof. 8. A kit according to claim 7 wherein said identifier comprises auxiliary plate information identifying an auxiliary plate for use with said test plate. 9. A kit according to claim 8 wherein said auxiliary plate information comprises auxiliary plate lot information. 10. A kit according to claim 8 wherein said auxiliary plate information comprises an auxiliary plate identification number. 11. A kit according to claim 1 wherein said plurality of auxiliary wells is a multiple of the number of assay wells in said assay test plate. 12. A kit according to claim 1 wherein said auxiliary plate comprises twice as many auxiliary wells as assay wells in said assay test plate. 13. A kit according to claim 1 wherein said auxiliary plate comprises four times as many auxiliary wells as assay wells in said assay test plate. 14. A kit according to claim 1 wherein said assay test plate further comprises a plurality of elements selected from the group consisting of a plate top, plate bottom, working electrodes, counter electrodes, reference electrodes, dielectric materials, electrical connections, dried or liquid assay reagents, and combinations thereof. 15. A kit according to claim 1 wherein said auxiliary plate further comprises a set of auxiliary wells, said set comprising adjacent auxiliary wells, wherein said set of auxiliary wells comprises reagents for an assay in a well of said assay test plate. 16. A kit according to claim 15 wherein said set comprises four adjacent auxiliary wells. 17. A kit according to claim 16 wherein said four adjacent auxiliary wells are arranged in a square. 18. A kit according to claim 16 wherein said four adjacent auxiliary wells are arranged in a row. 19. A kit according to claim 15 wherein a auxiliary well of said set of auxiliary wells is a dilution well. 20. A kit according to claim 15 wherein a auxiliary well of said set of auxiliary wells comprises pre-treated beads. 21. A kit according to claim 20 wherein said pre-treated beads are magnetic. 22. A kit according to claim 21 wherein said pre-treated beads comprise a coating selected from the group consisting of streptavidin, biotin, and avidin. 23. A kit according to claim 22 wherein one or more reagents in said set of auxiliary wells comprise a binding partner of said coating. 24. A kit according to claim 15 wherein at least one auxiliary well of said set comprises desiccant and the auxiliary plate comprises a seal. 25. A kit according to claim 24 wherein said at least one auxiliary well of said set is connected to an additional auxiliary well of said set via an air passage. 26. A kit according to claim 24 wherein said at least one auxiliary well of said set is connected to all auxiliary wells of said set via said air passage. 27. A kit according to claim 3 wherein said information is consumable information selected from the group consisting of lot identification information; lot specific analysis parameters, manufacturing process information, raw materials information, expiration date; calibration data; threshold information; the location of individual assay reagents and/or samples within one or more wells and/or auxiliary wells of said auxiliary plate and/or said test plate; Material Safety Data Sheet (MSDS) information, and combinations thereof. 28. A kit according to claim 3 wherein said information is sample information selected from the group consisting of the intended location of samples within said one or more wells of the test plate; assay results obtained on said test plate for said sample; identity of samples that have been and/or will be assayed in said test plate; and combinations thereof. 29. A kit according to claim 3 wherein said information is chain of custody information. 30. A kit according to claim 29 wherein said chain of custody information includes information regarding the control, transfer and/or analysis of a sample. 31. A kit according to claim 29 wherein said chain of custody information includes information regarding the control, transfer and/or analysis of a reagent. 32. A kit according to claim 29 wherein said information is chain of custody information regarding the control, transfer and/or manufacture of said auxiliary plate and/or test plate. 33. A kit according to claim 29 wherein said chain of custody information is selected from the group consisting of user identification; time and date stamp for said assay; location of an assay system using said auxiliary plate and test plate during said assay; calibration and QC status of said assay system during said assay, QC status of said auxiliary plate; QC status of said test plate, custody and/or location information for said auxiliary plate and/or test plate before and after the conduct of said assay; assay results for said sample; and combinations thereof. 34. A kit according to claim 29 wherein said information is chain of custody information selected from the group consisting of time, date, manufacturing personnel or processing parameters for one or more steps during the manufacture of said auxiliary plate and/or test plate; custody, location and or storage conditions for said auxiliary plate and/or test plate following manufacture and/or betweens steps during the manufacture of said auxiliary plate and/or test plate; and combinations thereof. 35. A kit according to claim 3 wherein said information is auxiliary plate and/or test plate information selected from the group consisting of plate type and structure; location and identity of assay reagents included with said auxiliary plate; location and identify of assay reagents included with said test plate; and combinations thereof. 36. A kit according to claim 3 wherein said assay is a multi-step assay and said information is assay process information that relates to a step or steps of said multi-step assay. 37. A kit according to claim 3 wherein said information is consumable security information selected from the group consisting of information concerning test plate and/or auxiliary plate authentication; information concerning defects in said test plate, auxiliary plate and/or a test site thereof; and combinations thereof. 38. A kit according to claim 1 wherein said test plate comprises capture antibodies for an agent selected from the group consisting of influenza-type A, influenza-type B, RSV, parainfluenza, and adenovirus; and said auxiliary plate comprises detection antibodies for said agent. 39. A kit according to claim 38 wherein said auxiliary plate further comprises desiccant. 40. A kit according to claim 1 wherein said test plate comprises capture antibodies for an agent selected from the group consisting of influenza-type A, influenza-type B, RSV, parainfluenza, adenovirus, influenza-type A (H1), influenza-type A (H2), influenza-type A (H3), influenza-type A (H5), influenza-type A (H7), influenza-type A (H9); and said auxiliary plate comprises one or more reagents selected from the group consisting of HA acidification buffer, HA neutralization buffer, a detection antibody for said agent, NP, and desiccant. 41. A kit according to claim 1 wherein said test plate comprises capture antibodies to a serum biomarker and said auxiliary plate comprises detection antibodies for said biomarker. 42. A kit according to claim 41 wherein said auxiliary plate further comprises desiccant. 43. An auxiliary plate comprising a plurality of assay auxiliary well, said auxiliary well comprising dry assay reagents for use in an assay with a corresponding assay test plate. 44. An auxiliary plate according to claim 43 wherein said auxiliary plate comprises an identifier comprising assay information used to identify an element selected from the group consisting of (i) said auxiliary plate, (ii) one or more auxiliary wells within said auxiliary plate, (iii) a reagent and/or sample that has been or will be used with said auxiliary plate, (iv) said test plate, (v) one or more wells within said test plate, (vi) a reagent and/or sample that has been or will be used with said test plate, and (vii) combinations thereof. 45. An auxiliary plate according to claim 44 wherein said identifier comprises test plate information identifying a test plate for use with said auxiliary plate. 46. An auxiliary plate according to claim 45 wherein said test plate information comprises test plate lot information. 47. An auxiliary plate according to claim 45 wherein said test plate information comprises a test plate identification number. 48. An auxiliary plate according to claim 43 wherein said plurality of auxiliary wells is a multiple of the number of wells in said assay test plate. 49. An auxiliary plate according to claim 48 wherein said auxiliary plate comprises twice as many auxiliary wells as wells in said assay test plate. 50. An auxiliary plate according to claim 48 wherein said auxiliary plate comprises four times as many auxiliary wells as wells in said assay test plate. 51. An auxiliary plate according to claim 43 wherein said auxiliary plate further comprises a set of auxiliary wells, said set comprising adjacent auxiliary wells, wherein said set of auxiliary wells comprises reagents for an assay in a well of said assay test plate. 52. An auxiliary plate according to claim 51 wherein said set comprises four adjacent auxiliary wells. 53. An auxiliary plate according to claim 52 wherein said four adjacent auxiliary wells are arranged in a square. 54. An auxiliary plate according to claim 52 wherein said four adjacent auxiliary wells are arranged in a row. 55. An auxiliary plate according to claim 51 wherein a auxiliary well of said set of auxiliary wells is a dilution well. 56. An auxiliary plate according to claim 51 wherein a auxiliary well of said set of auxiliary wells comprises pre-treated beads. 57. An auxiliary plate according to claim 56 wherein said pre-treated beads are magnetic. 58. An auxiliary plate according to claim 56 wherein said pre-treated beads comprise a coating selected from the group consisting of streptavidin, biotin, and avidin. 59. An auxiliary plate according to claim 58 wherein one or more reagents in said set of auxiliary wells comprise a binding partner of said coating. 60. An auxiliary plate according to claim 51 wherein at least one auxiliary well of said set comprises desiccant and said auxiliary plate comprises a seal. 61. An auxiliary plate according to claim 60 wherein said at least one auxiliary well of said set is connected to an additional auxiliary well of said set via an air passage. 62. An auxiliary plate according to claim 60 wherein said at least one auxiliary well of said set is connected to all auxiliary wells of said set via said air passage. 63. An auxiliary plate according to claim 43 wherein said information is consumable information selected from the group consisting of lot identification information; lot specific analysis parameters, manufacturing process information, raw materials information, expiration date; calibration data; threshold information; the location of individual assay reagents and/or samples within one or more wells and/or auxiliary wells of said auxiliary plate and/or said test plate; Material Safety Data Sheet (MSDS) information, and combinations thereof. 64. An auxiliary plate according to claim 43 wherein said information is sample information selected from the group consisting of the intended location of samples within said one or more auxiliary wells of the auxiliary plate; assay results obtained on said test plate for said sample; identity of samples that have been and/or will be assayed in said test plate; and combinations thereof. 65. An auxiliary plate according to claim 43 wherein said information is chain of custody information. 66. An auxiliary plate according to claim 65 wherein said chain of custody information includes information regarding the control, transfer and/or analysis of a reagent. 67. An auxiliary plate according to claim 65 wherein said information is chain of custody information regarding the control, transfer and/or manufacture of said auxiliary plate and/or test plate. 68. An auxiliary plate according to claim 65 wherein said chain of custody information is selected from the group consisting of user identification; time and date stamp for said assay; location of an assay system using said auxiliary plate and test plate during said assay; calibration and QC status of said assay system during said assay, QC status of said auxiliary plate; QC status of said test plate, custody and/or location information for said auxiliary plate and/or test plate before and after the conduct of said assay; assay results for said sample; and combinations thereof. 69. An auxiliary plate according to claim 65 wherein said information is chain of custody information selected from the group consisting of time, date, manufacturing personnel or processing parameters for one or more steps during the manufacture of said auxiliary plate and/or test plate; custody, location and or storage conditions for said auxiliary plate and/or test plate following manufacture and/or betweens steps during the manufacture of said auxiliary plate and/or test plate; and combinations thereof. 70. An auxiliary plate according to claim 43 wherein said information is auxiliary plate and/or test plate information selected from the group consisting of plate type and structure;
location and identity of assay reagents included with said auxiliary plate; location and identify of assay reagents included with said test plate; and combinations thereof. 71. An auxiliary plate according to claim 43 wherein said assay is a multi-step assay and said information is assay process information that relates to a step or step(s) of said multi-step assay. 72. An auxiliary plate according to claim 43 wherein said information is consumable security information selected from the group consisting of information concerning test plate and/or auxiliary plate authentication; information concerning defects in said test plate, auxiliary plate and/or a test well thereof; and combinations thereof. 73. An auxiliary plate according to claim 43 wherein said auxiliary plate comprises detection antibodies for an agent selected from the group consisting of influenza-type A, influenza-type B, RSV, Parainfluenza, and adenovirus. 74. An auxiliary plate according to claim 73 wherein said auxiliary plate further comprises desiccant. 75. An auxiliary plate according to claim 43 wherein said auxiliary plate comprises one or more reagents selected from the group consisting of HA acidification buffer, HA neutralization buffer, NP, desiccant, and a detection antibody for an agent selected from the group consisting of influenza-type A, influenza-type B, RSV, parainfluenza, adenovirus, influenza-type A (H1), influenza-type A (H2), influenza-type A (H3), influenza-type A (H5), influenza-type A (H7), influenza-type A (H9). 76. An auxiliary plate according to claim 43 wherein said auxiliary plate comprises detection antibodies to a serum biomarker. 77. An auxiliary plate according to claim 76 wherein said auxiliary plate further comprises desiccant. 78. An apparatus for conducting a measurement in a multi-well assay test plate, said apparatus comprising
(a) a subassembly capable of supporting and translating said test plate to one or more components of said apparatus; and (b) an auxiliary plate subassembly, wherein said auxiliary plate comprises a plurality of auxiliary wells comprising dry assay reagents for use in an assay with said test plate. 79. An apparatus according to claim 78 further comprising a pipettor subassembly that delivers sample and/or reagent to and from a well of said test plate and/or an auxiliary well of said auxiliary plate. 80. An apparatus according to claim 79 wherein said pipettor subassembly comprises a component selected from the group consisting of a pump, a plate piercing probe, a pipetting probe and an ultrasonic sensor. 81. An apparatus according to claim 78 wherein said subassembly capable of supporting and translating said test plate comprises a plate introduction aperture and a plate translation stage. 82. An apparatus according to claim 81 further comprising a plate stacker adjacent to said plate introduction aperture. 83. An apparatus according to claim 82 further comprising a plate elevator comprising a plate lifting platform that can be raised and lowered onto said plate translation stage. 84. An apparatus of claim 81 further comprising (i) an input plate introduction aperture comprising an input plate stacker and (ii) an output plate introduction aperture comprising an output plate stacker. 85. An apparatus of any one of claims 81 or 84 wherein said plate stacker can accommodate more than one test plate. 86. An apparatus of claim 81 wherein said subassembly is a light-tight enclosure and said plate introduction aperture comprises a sliding light-tight door. 87. An apparatus of claim 81 wherein said subassembly further comprises a component selected from the group consisting of a thermoelectric heater/cooler, a desiccant chamber, and an identifier controller. 88. An apparatus of claim 86 further comprising an imaging system mounted to an imaging aperture in the light-tight enclosure. 89. An apparatus of claim 81 wherein said plate translation stage is configured to position a well of a test plate in proximity to one or more components of said subassembly selected from the group consisting of said plate elevator, a well-wash subassembly, and an imaging system. 90. An apparatus of claim 89 wherein said well-wash subassembly comprises a seal removal tool, a well-wash head, a wash station, and fluidic connectors to a liquid reagent subassembly. 91. An apparatus of claim 90 wherein said well-wash head comprises a pipetting probe and a pipetting translation stage for translating said pipetting probe in a vertical direction. 92. An apparatus of claim 91 wherein said pipetting probe comprises a dispensing tube and a plurality of aspiration tubes. 93. An apparatus of claim 81 wherein said auxiliary plate subassembly comprises an auxiliary plate introduction aperture and a plate support. 94. An apparatus of claim 93 wherein said auxiliary plate subassembly further comprises a housing comprising two or more compartments, each compartment comprising said auxiliary plate introduction aperture and said plate support. 95. An apparatus of claim 94 wherein said compartments comprise a component selected from the group consisting of an identifier controller, a thermoelectric heater/cooler, and a desiccant chamber. 96. A method for conducting a measurement in a multi-well assay test plate, said method comprising the steps of:
(a) dispensing sample and/or reagent into an auxiliary well of an auxiliary plate, said auxiliary plate comprising a plurality of auxiliary wells, said plurality of auxiliary wells comprising dry assay reagents for use in an assay with said test plate; and (b) transferring sample and/or reagent from said auxiliary well to a well of a said assay test plate. 97. A method according to claim 96 wherein said dispensing step (a) comprises pre-treating said sample and/or reagent in said auxiliary well. 98. A method according to claim 97 wherein said transferring step (b) comprises dispensing pre-treated sample and/or reagent from said auxiliary well to said well of said test plate. 99. A method according to claim 96 wherein said assay test plate is supported on a plate translation stage and said method comprises translating said test plate via said plate translation stage to one or more components of said apparatus. 100. A method according to claim 99 wherein said method further comprises placing said assay test plate through a plate introduction aperture onto a plate stacker adjacent said plate translation stage. 101. A method according to claim 100 wherein said method further comprises lowering said assay test plate from said plate stacker to said plate translation stage. 102. A method according to claim 101 wherein said method further comprises (i) placing said assay test plate through an input plate introduction aperture comprising an input plate stacker, (ii) lowering said assay test plate from said input plate stacker to said plate translation stage, (iii) translating said assay test plate to one or more components of said apparatus to conduct said measurement, (iv) raising said assay test plate from said plate translation stage to an output plate stacker, and (v) removing said assay test plate from an output plate introduction aperture. 103. A method according to claim 96 wherein said method further comprises repeating steps (a) and (b) in an additional auxiliary well of said auxiliary plate and an additional test well of said test plate. 104. A method for conducting a measurement in a multi-well assay test plate, said method comprising the steps of:
(a) dispensing sample and/or reagent into an auxiliary well of a set of auxiliary wells in an auxiliary plate, said auxiliary plate comprising a plurality of auxiliary wells, said plurality of auxiliary wells comprising dry assay reagents for use in an assay with said test plate; (b) pre-treating said sample and/or reagent in one or more auxiliary wells of said set; (c) dispensing pre-treated sample and/or reagent from said one or more auxiliary wells of said set to a well of a said assay test plate; (d) repeating steps (a)-(c) with an additional sample and reagents in an additional set of auxiliary wells in said auxiliary plate, and an additional well of said test plate; (e) ejecting a used auxiliary plate; (f) repeating steps (a)-(c) with an additional auxiliary plate; and (g) ejecting a used assay test plate. 105. A well-wash subassembly comprising a multi-tube array comprising a central dispensing tube element surrounded by a plurality of aspiration tube elements. 106. A well-wash subassembly according to claim 105 wherein said multi-tube array comprises at least two dispensing tube elements at the center of the array. 107. A well-wash subassembly according to claim 106 wherein said dispensing tube elements comprise an independent fluid channel for buffers and/or diluents used during an assay. 108. A well-wash subassembly according to claim 105 wherein said aspiration tube elements surround the dispensing tube elements and said aspiration tube elements are positioned to align with the outer portions of a well bottom of a multi-well test plate. 109. A well-wash subassembly according to claim 105 wherein said aspiration tube elements are independently connected to dedicated fluidic lines. 110. A well-wash subassembly according to claim 105 wherein said multi-tube array comprises at least four aspiration tube elements. 111. An apparatus according to claim 80 wherein said pipettor subassembly comprises a reflective sensor comprising an infra-red LED and a phototransistor, wherein said LED and phototransistor are positioned in said subassembly to detect the presence or absence of a pipetting tip in said subassembly. | 1,600 |
1,032 | 16,062,154 | 1,612 | The disclosure describes compositions for dental varnishes, methods of making the compositions, and methods of using the compositions, such as in the treatment and prevention of hypersensitivity in teeth. | 1. A dental varnish composition comprising:
a fluoride source; a non-aqueous solvent; ethyl cellulose in an amount effective to achieve suspension of the fluoride source; and one or more resins selected from mastic, colophonium, or a combination thereof; wherein a high shear viscosity of the dental varnish composition is at least 1,500 centipoise (cps). 2. The dental varnish composition of claim 1, wherein the ethyl cellulose has a viscosity from 5 cps to 60 cps. 3. The dental varnish composition of claim 1, wherein the dental varnish composition has a high shear viscosity from 1,500 cps to 6,500 cps. 4. The dental varnish composition of claim 1, wherein the one or more resins include colophonium present in an amount from 28% to 38% and mastic present in an amount from 22% to 32%. 5. The dental varnish composition of claim 1, wherein the ethyl cellulose is present in an amount from 1% to 10%. 6. The dental varnish composition of claim 1, wherein the fluoride source is sodium fluoride and the fluoride source is present in an amount from 2% to 8%. 7. The dental varnish composition of claim 1, wherein the dental varnish composition has a ratio of yield stress as the shear rate is ramping up to yield stress when a shear rate is ramping down from 2.5 and 5.0. 8. The dental varnish composition of claim 1, wherein:
the fluoride source is present in an amount from 4% to 6%; the non-aqueous solvent is ethanol present in an amount from 24% to 30%; ethyl cellulose is present in an amount from 1% to 10%; mastic is present in an amount from 26% to 30%; and colophonium is present in an amount from 29% to 35%. 9. The dental varnish of claim 8, wherein the ethyl cellulose has a viscosity from 7 cps to 12 cps and is present in an amount of 3% to 8%. 10. The dental varnish of claim 8, wherein the ethyl cellulose has a viscosity from 17 cps to 25 cps and is present in an amount of 1% to 5%. 11. The dental varnish of claim 8, wherein the ethyl cellulose has a viscosity from 45 cps to 55 cps and is present in an amount of 1% to 5%. 12. The dental varnish composition of claim 1, further comprising beeswax present in an amount of 1% to 6%, a sweetener component present in an amount of 0.4% to 0.9%; and a flavoring component present in an amount of 0.4% to 0.9%. 13. The dental varnish composition of claim 1, wherein the dental varnish composition is free of shellac. 14. A dental varnish composition comprising:
a non-aqueous solvent; a fluoride source; ethyl cellulose in an amount effective to achieve suspension of the fluoride source, the ethyl cellulose having a viscosity no greater than 45 centipoise (cps); and one or more resins selected from mastic, colophonium, or a combination thereof. 15. The dental varnish composition of claim 14, wherein a high shear viscosity of the dental varnish composition is from 3,500 cps to 5,800 cps. 16. The dental varnish composition of claim 14, wherein a total amount of the one or more resins is from 42% to 70%. 17. The dental varnish composition of claim 14, wherein the ethyl cellulose has a viscosity from 8 cps to 11 cps and is present in an amount from 5% to 7% or the ethyl cellulose has a viscosity from 18 cps to 24 cps and is present in an amount from 2% to 4%. 18. A process comprising:
combining a first portion of an amount of an ethanol solution and a first portion of an amount of colophonium to form an ethanol solution and colophonium mixture; combining the ethanol solution and colophonium mixture with a fluoride source to form a first dental varnish component; combining a second portion of the amount of the ethanol solution, a second portion of the amount of the colophonium, a mastic, and an ethyl cellulose to form a second dental varnish component; and combining the first dental varnish component, the second dental varnish component, and a remainder of the amount of the ethanol solution to produce a composition of a dental varnish. 19. The process of claim 18, wherein:
the ethanol solution and colophonium mixture is mixed with the fluoride source for a period of time of 20 minutes to 50 minutes; the second portion of the amount of the ethanol solution, the second portion of the amount of the colophonium, the mastic, and the ethyl cellulose are mixed at a temperature from 100° C. to 125° C. for a duration from 1 hour to 6 hours; and the second dental varnish component is cooled to a temperature from 50° C. to 80° C. before being mixed with the first dental varnish component for a duration of 10 minutes to 40 minutes. 20. The process of claim 18, wherein:
the first portion of the amount of the ethanol solution is from 50% to 75% by weight of a total weight of the ethanol solution and colophonium mixture and the first portion of the amount of the colophonium is from 25% to 45% by weight of the total weight of the ethanol solution and colophonium mixture; an amount of the ethanol solution and colophonium mixture is from 50% to 75% by weight of a total weight of the first dental varnish component and an amount of the fluoride source is from 25% to 45% by weight of a total weight of the first dental varnish component; and the second portion of the amount of the ethanol solution is from 22% to 28% by weight of a total weight of the second dental varnish component, the second portion of the amount of the colophonium is from 20% to 45% by weight of the total weight of the second dental varnish component, an amount of the mastic is from 20% to 45% by weight of the total weight of the second dental varnish component, and an amount of the ethyl cellulose is from 1% to 10% of the second dental varnish component. | The disclosure describes compositions for dental varnishes, methods of making the compositions, and methods of using the compositions, such as in the treatment and prevention of hypersensitivity in teeth.1. A dental varnish composition comprising:
a fluoride source; a non-aqueous solvent; ethyl cellulose in an amount effective to achieve suspension of the fluoride source; and one or more resins selected from mastic, colophonium, or a combination thereof; wherein a high shear viscosity of the dental varnish composition is at least 1,500 centipoise (cps). 2. The dental varnish composition of claim 1, wherein the ethyl cellulose has a viscosity from 5 cps to 60 cps. 3. The dental varnish composition of claim 1, wherein the dental varnish composition has a high shear viscosity from 1,500 cps to 6,500 cps. 4. The dental varnish composition of claim 1, wherein the one or more resins include colophonium present in an amount from 28% to 38% and mastic present in an amount from 22% to 32%. 5. The dental varnish composition of claim 1, wherein the ethyl cellulose is present in an amount from 1% to 10%. 6. The dental varnish composition of claim 1, wherein the fluoride source is sodium fluoride and the fluoride source is present in an amount from 2% to 8%. 7. The dental varnish composition of claim 1, wherein the dental varnish composition has a ratio of yield stress as the shear rate is ramping up to yield stress when a shear rate is ramping down from 2.5 and 5.0. 8. The dental varnish composition of claim 1, wherein:
the fluoride source is present in an amount from 4% to 6%; the non-aqueous solvent is ethanol present in an amount from 24% to 30%; ethyl cellulose is present in an amount from 1% to 10%; mastic is present in an amount from 26% to 30%; and colophonium is present in an amount from 29% to 35%. 9. The dental varnish of claim 8, wherein the ethyl cellulose has a viscosity from 7 cps to 12 cps and is present in an amount of 3% to 8%. 10. The dental varnish of claim 8, wherein the ethyl cellulose has a viscosity from 17 cps to 25 cps and is present in an amount of 1% to 5%. 11. The dental varnish of claim 8, wherein the ethyl cellulose has a viscosity from 45 cps to 55 cps and is present in an amount of 1% to 5%. 12. The dental varnish composition of claim 1, further comprising beeswax present in an amount of 1% to 6%, a sweetener component present in an amount of 0.4% to 0.9%; and a flavoring component present in an amount of 0.4% to 0.9%. 13. The dental varnish composition of claim 1, wherein the dental varnish composition is free of shellac. 14. A dental varnish composition comprising:
a non-aqueous solvent; a fluoride source; ethyl cellulose in an amount effective to achieve suspension of the fluoride source, the ethyl cellulose having a viscosity no greater than 45 centipoise (cps); and one or more resins selected from mastic, colophonium, or a combination thereof. 15. The dental varnish composition of claim 14, wherein a high shear viscosity of the dental varnish composition is from 3,500 cps to 5,800 cps. 16. The dental varnish composition of claim 14, wherein a total amount of the one or more resins is from 42% to 70%. 17. The dental varnish composition of claim 14, wherein the ethyl cellulose has a viscosity from 8 cps to 11 cps and is present in an amount from 5% to 7% or the ethyl cellulose has a viscosity from 18 cps to 24 cps and is present in an amount from 2% to 4%. 18. A process comprising:
combining a first portion of an amount of an ethanol solution and a first portion of an amount of colophonium to form an ethanol solution and colophonium mixture; combining the ethanol solution and colophonium mixture with a fluoride source to form a first dental varnish component; combining a second portion of the amount of the ethanol solution, a second portion of the amount of the colophonium, a mastic, and an ethyl cellulose to form a second dental varnish component; and combining the first dental varnish component, the second dental varnish component, and a remainder of the amount of the ethanol solution to produce a composition of a dental varnish. 19. The process of claim 18, wherein:
the ethanol solution and colophonium mixture is mixed with the fluoride source for a period of time of 20 minutes to 50 minutes; the second portion of the amount of the ethanol solution, the second portion of the amount of the colophonium, the mastic, and the ethyl cellulose are mixed at a temperature from 100° C. to 125° C. for a duration from 1 hour to 6 hours; and the second dental varnish component is cooled to a temperature from 50° C. to 80° C. before being mixed with the first dental varnish component for a duration of 10 minutes to 40 minutes. 20. The process of claim 18, wherein:
the first portion of the amount of the ethanol solution is from 50% to 75% by weight of a total weight of the ethanol solution and colophonium mixture and the first portion of the amount of the colophonium is from 25% to 45% by weight of the total weight of the ethanol solution and colophonium mixture; an amount of the ethanol solution and colophonium mixture is from 50% to 75% by weight of a total weight of the first dental varnish component and an amount of the fluoride source is from 25% to 45% by weight of a total weight of the first dental varnish component; and the second portion of the amount of the ethanol solution is from 22% to 28% by weight of a total weight of the second dental varnish component, the second portion of the amount of the colophonium is from 20% to 45% by weight of the total weight of the second dental varnish component, an amount of the mastic is from 20% to 45% by weight of the total weight of the second dental varnish component, and an amount of the ethyl cellulose is from 1% to 10% of the second dental varnish component. | 1,600 |
1,033 | 15,061,488 | 1,619 | Pharmaceutical compositions for intraocular injection are described, the compositions consisting essentially of a therapeutically effective quantity of an anti-bacterial agent (such as moxifloxacin), a therapeutically effective quantity of an anti-inflammatory agent (such as prednisolone), at least one pharmaceutically acceptable excipient and a pharmaceutically acceptable carrier. Methods for fabricating the compositions and using them for intraocular injections are also described. | 1. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising delivering to the subject a composition comprising:
(a) a therapeutic component consisting essentially of:
(a1) a therapeutically effective quantity of an anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof; and
(a2) a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof;
(b) at least one pharmaceutically acceptable solubilizing and suspending agent selected from the group consisting of non-ionic polyoxyethylene-polyoxypropylene block copolymers; and (c) optionally, a pharmaceutically acceptable carrier therefor,
wherein the method of delivery is selected from the group consisting of intravitreal injection, intraocular intracameral injection, intra-lesional injection, intra-articular injection, subconjunctival injection, sub-tenon injection, delivery via eye drops, delivery via spray and intra-canalicular delivery, to treat the ophthalmological disease, condition or pathology thereby. 2. The method of claim 1, wherein the mammalian subject is selected from the group consisting of a human, a cat, a dog, another pet, a wild animal and a farm animal. 3. The method of claim 1, wherein the method of delivery is delivery via eye drops. 4. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising:
(a) performing a keratomileusis surgery on the subject; and (b) administering to the subject a composition comprising:
a. a therapeutic component consisting essentially of:
a1. a therapeutically effective quantity of an anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof; and
a2. a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof;
b. at least one pharmaceutically acceptable solubilizing and suspending agent selected from the group consisting of non-ionic polyoxyethylene-polyoxypropylene block copolymers; and
c. optionally, a pharmaceutically acceptable carrier therefor. 5. The method of claim 4, wherein the keratomileusis surgery is selected from the group consisting of laser-assisted in situ surgery (LASIK), photorefractive keratectomy (PRK), laser-assisted sub-epithelial keratectomy (LASEK), corneal ring segments, corneal cross linking, refractive corneal inlays and corneal lenticular surgery. 6. The method of claim 5, wherein the keratomileusis surgery is LASIK surgery. 7. The method of claim 4, wherein the composition is administered via drops after the surgery. | Pharmaceutical compositions for intraocular injection are described, the compositions consisting essentially of a therapeutically effective quantity of an anti-bacterial agent (such as moxifloxacin), a therapeutically effective quantity of an anti-inflammatory agent (such as prednisolone), at least one pharmaceutically acceptable excipient and a pharmaceutically acceptable carrier. Methods for fabricating the compositions and using them for intraocular injections are also described.1. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising delivering to the subject a composition comprising:
(a) a therapeutic component consisting essentially of:
(a1) a therapeutically effective quantity of an anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof; and
(a2) a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof;
(b) at least one pharmaceutically acceptable solubilizing and suspending agent selected from the group consisting of non-ionic polyoxyethylene-polyoxypropylene block copolymers; and (c) optionally, a pharmaceutically acceptable carrier therefor,
wherein the method of delivery is selected from the group consisting of intravitreal injection, intraocular intracameral injection, intra-lesional injection, intra-articular injection, subconjunctival injection, sub-tenon injection, delivery via eye drops, delivery via spray and intra-canalicular delivery, to treat the ophthalmological disease, condition or pathology thereby. 2. The method of claim 1, wherein the mammalian subject is selected from the group consisting of a human, a cat, a dog, another pet, a wild animal and a farm animal. 3. The method of claim 1, wherein the method of delivery is delivery via eye drops. 4. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising:
(a) performing a keratomileusis surgery on the subject; and (b) administering to the subject a composition comprising:
a. a therapeutic component consisting essentially of:
a1. a therapeutically effective quantity of an anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof; and
a2. a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof;
b. at least one pharmaceutically acceptable solubilizing and suspending agent selected from the group consisting of non-ionic polyoxyethylene-polyoxypropylene block copolymers; and
c. optionally, a pharmaceutically acceptable carrier therefor. 5. The method of claim 4, wherein the keratomileusis surgery is selected from the group consisting of laser-assisted in situ surgery (LASIK), photorefractive keratectomy (PRK), laser-assisted sub-epithelial keratectomy (LASEK), corneal ring segments, corneal cross linking, refractive corneal inlays and corneal lenticular surgery. 6. The method of claim 5, wherein the keratomileusis surgery is LASIK surgery. 7. The method of claim 4, wherein the composition is administered via drops after the surgery. | 1,600 |
1,034 | 15,057,711 | 1,619 | Pharmaceutical ophthalmic compositions are described, the compositions consisting essentially of a therapeutically effective quantity of an anti-bacterial agent (such as moxifloxacin), a therapeutically effective quantity of an anti-inflammatory agent (such as prednisolone), at least one pharmaceutically acceptable excipient and a pharmaceutically acceptable carrier. Methods for fabricating the compositions and using them are also described. | 1. A pharmaceutical composition formulated as a suspension that consists of:
(a) a dispersed phase consisting of solid particles consisting of a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof, and pharmaceutically acceptable salts, hydrates and solvates thereof; and (b) a dispersion medium consisting of:
(b1) a therapeutically effective quantity of at least one anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof;
(b2) a therapeutically effective quantity of at least one pharmaceutically acceptable solubilizing and suspending agent selected from the group consisting of non-ionic polyoxyethlene-polyoxypropylene block copolymers and polysorbates;
(b3) optionally, a therapeutically effective quantity of at least one glycopeptide antibiotic selected from the group consisting of vancomycin, teicoplanin, telavancin, decaplanin, ramoplanin, gentamicin, tobramycin, amikacin, cefuroxime, polymyxin B sulfate, and trimethoprim;
(b4) optionally, a therapeutically effective quantity of at least one non-steroid anti-inflammatory drug selected from the group consisting of bromfenac, ketorolac, etodolac, sulindac, diclofenac, aceclofenac, nepafenac, tolmetin, indomethacin, nabumetone, ketoprofen, dexketoprofen, ibuprofen, flurbiprofen, dexibuprofen, fenoprofen, loxoprofen, oxaprozin, naproxen, aspirin, salicylic acid, diflunisal, salsalate, mefenamic acid, meclofenamic acid, flufenamic acid, tolfenamic acid, meloxicam, piroxicam, ternoxicam, droxicam, lornoxicam, isoxicam, celecoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, firocoxib, nimesulide, clonixin, licofelone, and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof; and
(b5) a pharmaceutically acceptable carrier,
wherein the dispersed phase is dispersed within the dispersion medium. 2. The pharmaceutical composition of claim 1, wherein the corticosteroid is selected from the group consisting of prednisone, prednisolone, methylprednisone and derivatives or analogs thereof. 3. The pharmaceutical composition of claim 1, wherein the anti-bacterial agent is a fluorinated quinolone. 4. The pharmaceutical composition of claim 3, wherein the anti-bacterial agent has the chemical structure A: 5. The pharmaceutical composition of claim 3, wherein the fluorinated quinolone is selected from the group consisting of moxifloxacin and gatifloxacin. 6. The pharmaceutical composition of claim 5, wherein the fluorinated quinolone is moxifloxacin. 7. The pharmaceutical composition of claim 1, wherein the non-ionic polyoxyethlene-polyoxypropylene block copolymer is poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol). 8. The pharmaceutical composition of claim 7, wherein:
(a) the anti-bacterial agent is moxifloxacin at a concentration of about 1.0 mg/mL; (b) the corticosteroid is prednisone at a concentration of about 15.0 mg/mL; and (c) the non-ionic polyoxyethlene-polyoxypropylene block copolymer is at a concentration of about 5.0 mass %. 9. The pharmaceutical composition of claim 1, wherein about 99% of all the solid particles in the dispersed phase have the diameter of 5 μM or less. 10. The pharmaceutical composition of claim 9, wherein more than 80% of the solid particles have sizes within the range between about 1 μM and about 4 μM. 11. The pharmaceutical composition of claim 1, wherein the glycopeptide antibiotic, if present, is vancomycin. 12. The pharmaceutical composition of claim 1, wherein the non-steroid anti-inflammatory drug, if present, is bromfenac. 13. A method for preparing a pharmaceutical composition comprising combining components (a) and (b) of claim 1, to obtain the pharmaceutical composition thereby. 14. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising delivery to the subject the composition of claim 1, wherein the method of delivery is selected from the group consisting of intravitreal injection, intraocular intracameral injection, intra-lesional injection, intra-articular injection, subconjunctival injection, sub-tenon injection, delivery via eye drops, delivery via spray and intra-canalicular delivery, to treat the ophthalmological disease, condition or pathology thereby. 15. The method of claim 14, wherein the mammalian subject is selected from the group consisting of a human, a cat, a dog, another pet, a wild animal and a farm animal. 16. The method of claim 14, wherein the method of delivery is delivery via eye drops. 17. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising:
(a) performing a keratomileusis surgery on the subject; and (b) administering to the subject the composition of claim 1. 18. The method of claim 17, wherein the keratomileusis surgery is selected from the group consisting of laser-assisted in situ surgery (LASIK), photorefractive keratectomy (PRK), laser-assisted sub-epithelial keratectomy (LASEK), corneal ring segments, corneal cross linking, refractive corneal inlays and corneal lenticular surgery. 19. The method of claim 18, wherein the keratomileusis surgery is LASIK surgery. 20. The method of claim 17, wherein the composition is administered via drops after the surgery. | Pharmaceutical ophthalmic compositions are described, the compositions consisting essentially of a therapeutically effective quantity of an anti-bacterial agent (such as moxifloxacin), a therapeutically effective quantity of an anti-inflammatory agent (such as prednisolone), at least one pharmaceutically acceptable excipient and a pharmaceutically acceptable carrier. Methods for fabricating the compositions and using them are also described.1. A pharmaceutical composition formulated as a suspension that consists of:
(a) a dispersed phase consisting of solid particles consisting of a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof, and pharmaceutically acceptable salts, hydrates and solvates thereof; and (b) a dispersion medium consisting of:
(b1) a therapeutically effective quantity of at least one anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof;
(b2) a therapeutically effective quantity of at least one pharmaceutically acceptable solubilizing and suspending agent selected from the group consisting of non-ionic polyoxyethlene-polyoxypropylene block copolymers and polysorbates;
(b3) optionally, a therapeutically effective quantity of at least one glycopeptide antibiotic selected from the group consisting of vancomycin, teicoplanin, telavancin, decaplanin, ramoplanin, gentamicin, tobramycin, amikacin, cefuroxime, polymyxin B sulfate, and trimethoprim;
(b4) optionally, a therapeutically effective quantity of at least one non-steroid anti-inflammatory drug selected from the group consisting of bromfenac, ketorolac, etodolac, sulindac, diclofenac, aceclofenac, nepafenac, tolmetin, indomethacin, nabumetone, ketoprofen, dexketoprofen, ibuprofen, flurbiprofen, dexibuprofen, fenoprofen, loxoprofen, oxaprozin, naproxen, aspirin, salicylic acid, diflunisal, salsalate, mefenamic acid, meclofenamic acid, flufenamic acid, tolfenamic acid, meloxicam, piroxicam, ternoxicam, droxicam, lornoxicam, isoxicam, celecoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, firocoxib, nimesulide, clonixin, licofelone, and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof; and
(b5) a pharmaceutically acceptable carrier,
wherein the dispersed phase is dispersed within the dispersion medium. 2. The pharmaceutical composition of claim 1, wherein the corticosteroid is selected from the group consisting of prednisone, prednisolone, methylprednisone and derivatives or analogs thereof. 3. The pharmaceutical composition of claim 1, wherein the anti-bacterial agent is a fluorinated quinolone. 4. The pharmaceutical composition of claim 3, wherein the anti-bacterial agent has the chemical structure A: 5. The pharmaceutical composition of claim 3, wherein the fluorinated quinolone is selected from the group consisting of moxifloxacin and gatifloxacin. 6. The pharmaceutical composition of claim 5, wherein the fluorinated quinolone is moxifloxacin. 7. The pharmaceutical composition of claim 1, wherein the non-ionic polyoxyethlene-polyoxypropylene block copolymer is poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol). 8. The pharmaceutical composition of claim 7, wherein:
(a) the anti-bacterial agent is moxifloxacin at a concentration of about 1.0 mg/mL; (b) the corticosteroid is prednisone at a concentration of about 15.0 mg/mL; and (c) the non-ionic polyoxyethlene-polyoxypropylene block copolymer is at a concentration of about 5.0 mass %. 9. The pharmaceutical composition of claim 1, wherein about 99% of all the solid particles in the dispersed phase have the diameter of 5 μM or less. 10. The pharmaceutical composition of claim 9, wherein more than 80% of the solid particles have sizes within the range between about 1 μM and about 4 μM. 11. The pharmaceutical composition of claim 1, wherein the glycopeptide antibiotic, if present, is vancomycin. 12. The pharmaceutical composition of claim 1, wherein the non-steroid anti-inflammatory drug, if present, is bromfenac. 13. A method for preparing a pharmaceutical composition comprising combining components (a) and (b) of claim 1, to obtain the pharmaceutical composition thereby. 14. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising delivery to the subject the composition of claim 1, wherein the method of delivery is selected from the group consisting of intravitreal injection, intraocular intracameral injection, intra-lesional injection, intra-articular injection, subconjunctival injection, sub-tenon injection, delivery via eye drops, delivery via spray and intra-canalicular delivery, to treat the ophthalmological disease, condition or pathology thereby. 15. The method of claim 14, wherein the mammalian subject is selected from the group consisting of a human, a cat, a dog, another pet, a wild animal and a farm animal. 16. The method of claim 14, wherein the method of delivery is delivery via eye drops. 17. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising:
(a) performing a keratomileusis surgery on the subject; and (b) administering to the subject the composition of claim 1. 18. The method of claim 17, wherein the keratomileusis surgery is selected from the group consisting of laser-assisted in situ surgery (LASIK), photorefractive keratectomy (PRK), laser-assisted sub-epithelial keratectomy (LASEK), corneal ring segments, corneal cross linking, refractive corneal inlays and corneal lenticular surgery. 19. The method of claim 18, wherein the keratomileusis surgery is LASIK surgery. 20. The method of claim 17, wherein the composition is administered via drops after the surgery. | 1,600 |
1,035 | 15,148,574 | 1,619 | Pharmaceutical ophthalmic compositions are described, the compositions consisting essentially of a therapeutically effective quantity of an anti-bacterial agent (such as moxifloxacin), a therapeutically effective quantity of an anti-inflammatory agent (such as prednisolone), a combination of at least two solubilizing and suspending agents (of which one is a non-ionic polyoxyethlene-polyoxypropylene block copolymer), and a carrier. Methods for fabricating the compositions and using them are also described. | 1. A pharmaceutical composition formulated as a suspension that consists of:
(a) a dispersed phase consisting of solid particles consisting of a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof, and pharmaceutically acceptable salts, hydrates and solvates thereof; and (b) a dispersion medium consisting of:
(b1) a therapeutically effective quantity of at least one anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof;
(b2) a therapeutically effective quantity of a combination of at least two pharmaceutically acceptable solubilizing and suspending agents, the combination consisting of:
(b2a) a first solubilizing and suspending agent selected from the group consisting of at least one non-ionic polyoxyethlene-polyoxypropylene block copolymer; and
(b2b) a second solubilizing and suspending agent selected from the group consisting of a water-soluble derivative of cellulose, optionally partially cross-linked polyacrylates, polyoxyethylene sorbitan monolaurates, polyoxyethylene sorbitan monopalmitates, polyoxyethylene sorbitan monostearates, polyoxyethylene sorbitan monooleates or combinations thereof;
(b3) optionally, a therapeutically effective quantity of at least one glycopeptide antibiotic selected from the group consisting of vancomycin, teicoplanin, telavancin, decaplanin, ramoplanin, gentamicin, tobramycin, amikacin, cefuroxime, polymyxin B sulfate, and trimethoprim;
(b4) optionally, a therapeutically effective quantity of at least one non-steroid anti-inflammatory drug selected from the group consisting of bromfenac, ketorolac, etodolac, sulindac, diclofenac, aceclofenac, nepafenac, tolmetin, indomethacin, nabumetone, ketoprofen, dexketoprofen, ibuprofen, flurbiprofen, dexibuprofen, fenoprofen, loxoprofen, oxaprozin, naproxen, aspirin, salicylic acid, diflunisal, salsalate, mefenamic acid, meclofenamic acid, flufenamic acid, tolfenamic acid, meloxicam, piroxicam, ternoxicam, droxicam, lornoxicam, isoxicam, celecoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, firocoxib, nimesulide, clonixin, licofelone, and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof; and
(b5) a pharmaceutically acceptable carrier,
wherein the dispersed phase is dispersed within the dispersion medium, with the further proviso that the pharmaceutical composition is an ophthalmic composition that is suitable for delivery via intraocular injection or via eye drops. 2. The pharmaceutical composition of claim 1, wherein the corticosteroid is selected from the group consisting of prednisone, prednisolone, methylprednisone and derivatives or analogs thereof. 3. The pharmaceutical composition of claim 1, wherein the anti-bacterial agent is a fluorinated quinolone. 4. The pharmaceutical composition of claim 3, wherein the anti-bacterial agent has the chemical structure A: 5. The pharmaceutical composition of claim 3, wherein the fluorinated quinolone is selected from the group consisting of moxifloxacin and gatifloxacin. 6. The pharmaceutical composition of claim 5, wherein the fluorinated quinolone is moxifloxacin. 7. The pharmaceutical composition of claim 1, wherein the non-ionic polyoxyethlene-polyoxypropylene block copolymer is poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol). 8. The pharmaceutical composition of claim 1, wherein the water-soluble derivative of cellulose is selected from the group consisting of carboxymethyl cellulose, methyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose. 9. The pharmaceutical composition of claim 1, wherein the second solubilizing and suspending agent is polyoxyethylene (20) sorbitan monooleate. 10. The pharmaceutical composition of claim 7, comprising:
(a) moxifloxacin at a concentration of about 1.0 mg/mL; (b) prednisone at a concentration of about 15.0 mg/mL; and (c) the non-ionic polyoxyethlene-polyoxypropylene block copolymer is at a concentration of about 5.0 mass %. 11. The pharmaceutical composition of claim 1, wherein the composition is a suspension comprising particles formed by component (a), wherein about 99% of all the particles have the diameter of 5 μM or less. 12. The pharmaceutical composition of claim 11, wherein more than 80% of the particles have the sizes within the range between about 1 μM and about 4 μM. 13. The pharmaceutical composition of claim 1, wherein the glycopeptide antibiotic, if present, is vancomycin. 14. The pharmaceutical composition of claim 1, wherein the non-steroid anti-inflammatory drug, if present, is bromfenac. 15. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising delivery to the subject the composition of claim 1, wherein the method of delivery is selected from the group consisting of intravitreal injection, intraocular intracameral injection, intra-lesional injection, intra-articular injection, subconjunctival injection, sub-tenon injection, delivery via eye drops, delivery via spray and intra-canalicular delivery, to treat the ophthalmological disease, condition or pathology thereby. 16. The method of claim 15, wherein the method of delivery is delivery via eye drops. 17. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising:
(a) performing a keratomileusis surgery on the subject; and (b) administering to the subject a composition of claim 1. 18. The method of claim 17, wherein the keratomileusis surgery is selected from the group consisting of laser-assisted in situ surgery (LASIK), photorefractive keratectomy (PRK), laser-assisted sub-epithelial keratectomy (LASEK), corneal ring segments, corneal cross linking, refractive corneal inlays and corneal lenticular surgery. 19. The method of claim 18, wherein the keratomileusis surgery is LASIK surgery. 20. The method of claim 17, wherein the composition is administered via drops after the surgery. | Pharmaceutical ophthalmic compositions are described, the compositions consisting essentially of a therapeutically effective quantity of an anti-bacterial agent (such as moxifloxacin), a therapeutically effective quantity of an anti-inflammatory agent (such as prednisolone), a combination of at least two solubilizing and suspending agents (of which one is a non-ionic polyoxyethlene-polyoxypropylene block copolymer), and a carrier. Methods for fabricating the compositions and using them are also described.1. A pharmaceutical composition formulated as a suspension that consists of:
(a) a dispersed phase consisting of solid particles consisting of a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof, and pharmaceutically acceptable salts, hydrates and solvates thereof; and (b) a dispersion medium consisting of:
(b1) a therapeutically effective quantity of at least one anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof;
(b2) a therapeutically effective quantity of a combination of at least two pharmaceutically acceptable solubilizing and suspending agents, the combination consisting of:
(b2a) a first solubilizing and suspending agent selected from the group consisting of at least one non-ionic polyoxyethlene-polyoxypropylene block copolymer; and
(b2b) a second solubilizing and suspending agent selected from the group consisting of a water-soluble derivative of cellulose, optionally partially cross-linked polyacrylates, polyoxyethylene sorbitan monolaurates, polyoxyethylene sorbitan monopalmitates, polyoxyethylene sorbitan monostearates, polyoxyethylene sorbitan monooleates or combinations thereof;
(b3) optionally, a therapeutically effective quantity of at least one glycopeptide antibiotic selected from the group consisting of vancomycin, teicoplanin, telavancin, decaplanin, ramoplanin, gentamicin, tobramycin, amikacin, cefuroxime, polymyxin B sulfate, and trimethoprim;
(b4) optionally, a therapeutically effective quantity of at least one non-steroid anti-inflammatory drug selected from the group consisting of bromfenac, ketorolac, etodolac, sulindac, diclofenac, aceclofenac, nepafenac, tolmetin, indomethacin, nabumetone, ketoprofen, dexketoprofen, ibuprofen, flurbiprofen, dexibuprofen, fenoprofen, loxoprofen, oxaprozin, naproxen, aspirin, salicylic acid, diflunisal, salsalate, mefenamic acid, meclofenamic acid, flufenamic acid, tolfenamic acid, meloxicam, piroxicam, ternoxicam, droxicam, lornoxicam, isoxicam, celecoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib, etoricoxib, firocoxib, nimesulide, clonixin, licofelone, and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof; and
(b5) a pharmaceutically acceptable carrier,
wherein the dispersed phase is dispersed within the dispersion medium, with the further proviso that the pharmaceutical composition is an ophthalmic composition that is suitable for delivery via intraocular injection or via eye drops. 2. The pharmaceutical composition of claim 1, wherein the corticosteroid is selected from the group consisting of prednisone, prednisolone, methylprednisone and derivatives or analogs thereof. 3. The pharmaceutical composition of claim 1, wherein the anti-bacterial agent is a fluorinated quinolone. 4. The pharmaceutical composition of claim 3, wherein the anti-bacterial agent has the chemical structure A: 5. The pharmaceutical composition of claim 3, wherein the fluorinated quinolone is selected from the group consisting of moxifloxacin and gatifloxacin. 6. The pharmaceutical composition of claim 5, wherein the fluorinated quinolone is moxifloxacin. 7. The pharmaceutical composition of claim 1, wherein the non-ionic polyoxyethlene-polyoxypropylene block copolymer is poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol). 8. The pharmaceutical composition of claim 1, wherein the water-soluble derivative of cellulose is selected from the group consisting of carboxymethyl cellulose, methyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose. 9. The pharmaceutical composition of claim 1, wherein the second solubilizing and suspending agent is polyoxyethylene (20) sorbitan monooleate. 10. The pharmaceutical composition of claim 7, comprising:
(a) moxifloxacin at a concentration of about 1.0 mg/mL; (b) prednisone at a concentration of about 15.0 mg/mL; and (c) the non-ionic polyoxyethlene-polyoxypropylene block copolymer is at a concentration of about 5.0 mass %. 11. The pharmaceutical composition of claim 1, wherein the composition is a suspension comprising particles formed by component (a), wherein about 99% of all the particles have the diameter of 5 μM or less. 12. The pharmaceutical composition of claim 11, wherein more than 80% of the particles have the sizes within the range between about 1 μM and about 4 μM. 13. The pharmaceutical composition of claim 1, wherein the glycopeptide antibiotic, if present, is vancomycin. 14. The pharmaceutical composition of claim 1, wherein the non-steroid anti-inflammatory drug, if present, is bromfenac. 15. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising delivery to the subject the composition of claim 1, wherein the method of delivery is selected from the group consisting of intravitreal injection, intraocular intracameral injection, intra-lesional injection, intra-articular injection, subconjunctival injection, sub-tenon injection, delivery via eye drops, delivery via spray and intra-canalicular delivery, to treat the ophthalmological disease, condition or pathology thereby. 16. The method of claim 15, wherein the method of delivery is delivery via eye drops. 17. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising:
(a) performing a keratomileusis surgery on the subject; and (b) administering to the subject a composition of claim 1. 18. The method of claim 17, wherein the keratomileusis surgery is selected from the group consisting of laser-assisted in situ surgery (LASIK), photorefractive keratectomy (PRK), laser-assisted sub-epithelial keratectomy (LASEK), corneal ring segments, corneal cross linking, refractive corneal inlays and corneal lenticular surgery. 19. The method of claim 18, wherein the keratomileusis surgery is LASIK surgery. 20. The method of claim 17, wherein the composition is administered via drops after the surgery. | 1,600 |
1,036 | 14,972,822 | 1,619 | Pharmaceutical compositions for intraocular injection are described, the compositions consisting essentially of a therapeutically effective quantity of an anti-bacterial agent (such as moxifloxacin), a therapeutically effective quantity of an anti-inflammatory agent (such as prednisolone), at least one pharmaceutically acceptable excipient and a pharmaceutically acceptable carrier. Methods for fabricating the compositions and using them for intraocular injections are also described. | 1. A pharmaceutical composition, comprising:
(a) a therapeutic component consisting essentially of:
(a1) a therapeutically effective quantity of an anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof; and
(a2) a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof;
(b) at least one pharmaceutically acceptable solubilizing and suspending agent selected from the group consisting of non-ionic polyoxyethlene-polyoxypropylene block copolymers; and (c) optionally, a pharmaceutically acceptable carrier therefor,
wherein the composition is free of preservatives. 2. The pharmaceutical composition of claim 1, wherein the corticosteroid is selected from the group consisting of prednisone, prednisolone, methylprednisone and derivatives or analogs thereof. 3. The pharmaceutical composition of claim 1, wherein the anti-bacterial agent is a fluorinated quinolone. 4. The pharmaceutical composition of claim 3, wherein the anti-bacterial agent has the chemical structure A: 5. The pharmaceutical composition of claim 3, wherein the fluorinated quinolone is selected from the group consisting of moxifloxacin and gatifloxacin. 6. The pharmaceutical composition of claim 5, wherein the fluorinated quinolone is moxifloxacin. 7. The pharmaceutical composition of claim 1, wherein the non-ionic polyoxyethlene-polyoxypropylene block copolymer is Poloxamer 407®. 8. The pharmaceutical composition of claim 7, comprising:
(a) moxifloxacin at a concentration of about 1.0 mg/mL; (b) prednisone at a concentration of about 15.0 mg/mL; and (c) Poloxamer 407® at a concentration of about 5.0 mass %. 9. The pharmaceutical composition of claim 1, further comprising a therapeutically effective quantity of an antibiotic selected from the group consisting of vancomycin, teicoplanin, telavancin, decaplanin, ramoplanin, gentamicin, tobramycin, amikacin, cefuroxime, polymyxin B sulfate, trimethoprim, and a combination thereof. 10. The pharmaceutical composition of claim 9, wherein the antibiotic is vancomycin. 11. The pharmaceutical composition of claim 1, wherein the composition is a suspension comprising particles formed by components (a), (b) and (c), wherein about 99% of all the particles have the diameter of 5 μM or less. 12. The pharmaceutical composition of claim 11, wherein more than 80% of the particles have diameters within the range of between about 1 μM and about 4 μM. 13. A method for preparing a pharmaceutical composition comprising combining components (a), (b) and (c) of claim 1, to obtain the pharmaceutical composition thereby. 14. The method of claim 13, wherein the anti-bacterial agent is a fluorinated quinolone. 15. The method of claim 14, wherein the fluorinated quinolone is moxifloxacin. 16. The method of claim 13, wherein the corticosteroid is prednisone or a derivative thereof. 17. The method of claim 13, wherein:
(a) the anti-bacterial agent is moxifloxacin; and (b) the corticosteroid is prednisone or a derivative thereof. 18. The method of claim 17, wherein the non-ionic polyoxyethlene-polyoxypropylene block copolymer is Poloxamer 407®. 19. The method of claim 17, further comprising a therapeutically effective quantity of vancomycin. 20. The method of claim 13, wherein the anti-bacterial agent, the anti-inflammatory agent, the excipient and the carrier are combined in a one-batch formulation method. 21. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment, comprising delivering to the subject the composition of claim 1, wherein the method of delivery is selected from the group consisting of intravitreal injection, intraocular intracameral injection, intra-lesional injection, intra-articular injection, subconjunctival injection, sub-tenon injection, delivery via eye drops, delivery via spray and intra-canalicular delivery, to treat the ophthalmological disease, condition or pathology thereby. 22. The method of claim 21, wherein the mammalian subject is selected from the group consisting of a human, a cats, a dog, another pet, a wild animal and a farm animal. 23. The method of claim 21, wherein the method of delivery is delivery via eye drops. 24. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising:
(a) performing a keratomileusis surgery on the subject; and (b) administering to the subject a composition of claim 1. 25. The method of claim 24, wherein the keratomileusis surgery is selected from the group consisting of laser-assisted in situ surgery (LASIK), photorefractive keratectomy (PRK), laser-assisted sub-epithelial keratectomy (LASEK), corneal ring segments, corneal cross linking, refractive corneal inlays, and corneal lenticular surgery. 26. The method of claim 25, wherein the keratomileusis surgery is LASIK surgery. 27. The method of claim 24, wherein the composition is administered via drops after the surgery. 28. A pharmaceutical kit, comprising a sealed container containing the pharmaceutical composition of claim 1, and an instruction for use of the composition enclosed with the container. | Pharmaceutical compositions for intraocular injection are described, the compositions consisting essentially of a therapeutically effective quantity of an anti-bacterial agent (such as moxifloxacin), a therapeutically effective quantity of an anti-inflammatory agent (such as prednisolone), at least one pharmaceutically acceptable excipient and a pharmaceutically acceptable carrier. Methods for fabricating the compositions and using them for intraocular injections are also described.1. A pharmaceutical composition, comprising:
(a) a therapeutic component consisting essentially of:
(a1) a therapeutically effective quantity of an anti-bacterial agent independently selected from the group consisting of quinolone, a fluorinated quinolone and pharmaceutically acceptable salts, hydrates, solvates or N-oxides thereof; and
(a2) a therapeutically effective quantity of a corticosteroid independently selected from the group consisting of prednisone, prednisolone, methylprednisone, corticol, cortisone, fluorocortisone, deoxycorticosterone acetate, aldosterone, budesonide, derivatives or analogs thereof and pharmaceutically acceptable salts, hydrates, solvates, ethers, esters, acetals and ketals thereof;
(b) at least one pharmaceutically acceptable solubilizing and suspending agent selected from the group consisting of non-ionic polyoxyethlene-polyoxypropylene block copolymers; and (c) optionally, a pharmaceutically acceptable carrier therefor,
wherein the composition is free of preservatives. 2. The pharmaceutical composition of claim 1, wherein the corticosteroid is selected from the group consisting of prednisone, prednisolone, methylprednisone and derivatives or analogs thereof. 3. The pharmaceutical composition of claim 1, wherein the anti-bacterial agent is a fluorinated quinolone. 4. The pharmaceutical composition of claim 3, wherein the anti-bacterial agent has the chemical structure A: 5. The pharmaceutical composition of claim 3, wherein the fluorinated quinolone is selected from the group consisting of moxifloxacin and gatifloxacin. 6. The pharmaceutical composition of claim 5, wherein the fluorinated quinolone is moxifloxacin. 7. The pharmaceutical composition of claim 1, wherein the non-ionic polyoxyethlene-polyoxypropylene block copolymer is Poloxamer 407®. 8. The pharmaceutical composition of claim 7, comprising:
(a) moxifloxacin at a concentration of about 1.0 mg/mL; (b) prednisone at a concentration of about 15.0 mg/mL; and (c) Poloxamer 407® at a concentration of about 5.0 mass %. 9. The pharmaceutical composition of claim 1, further comprising a therapeutically effective quantity of an antibiotic selected from the group consisting of vancomycin, teicoplanin, telavancin, decaplanin, ramoplanin, gentamicin, tobramycin, amikacin, cefuroxime, polymyxin B sulfate, trimethoprim, and a combination thereof. 10. The pharmaceutical composition of claim 9, wherein the antibiotic is vancomycin. 11. The pharmaceutical composition of claim 1, wherein the composition is a suspension comprising particles formed by components (a), (b) and (c), wherein about 99% of all the particles have the diameter of 5 μM or less. 12. The pharmaceutical composition of claim 11, wherein more than 80% of the particles have diameters within the range of between about 1 μM and about 4 μM. 13. A method for preparing a pharmaceutical composition comprising combining components (a), (b) and (c) of claim 1, to obtain the pharmaceutical composition thereby. 14. The method of claim 13, wherein the anti-bacterial agent is a fluorinated quinolone. 15. The method of claim 14, wherein the fluorinated quinolone is moxifloxacin. 16. The method of claim 13, wherein the corticosteroid is prednisone or a derivative thereof. 17. The method of claim 13, wherein:
(a) the anti-bacterial agent is moxifloxacin; and (b) the corticosteroid is prednisone or a derivative thereof. 18. The method of claim 17, wherein the non-ionic polyoxyethlene-polyoxypropylene block copolymer is Poloxamer 407®. 19. The method of claim 17, further comprising a therapeutically effective quantity of vancomycin. 20. The method of claim 13, wherein the anti-bacterial agent, the anti-inflammatory agent, the excipient and the carrier are combined in a one-batch formulation method. 21. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment, comprising delivering to the subject the composition of claim 1, wherein the method of delivery is selected from the group consisting of intravitreal injection, intraocular intracameral injection, intra-lesional injection, intra-articular injection, subconjunctival injection, sub-tenon injection, delivery via eye drops, delivery via spray and intra-canalicular delivery, to treat the ophthalmological disease, condition or pathology thereby. 22. The method of claim 21, wherein the mammalian subject is selected from the group consisting of a human, a cats, a dog, another pet, a wild animal and a farm animal. 23. The method of claim 21, wherein the method of delivery is delivery via eye drops. 24. A method for treating an ophthalmological disease, condition or pathology in a mammalian subject in need of such treatment comprising:
(a) performing a keratomileusis surgery on the subject; and (b) administering to the subject a composition of claim 1. 25. The method of claim 24, wherein the keratomileusis surgery is selected from the group consisting of laser-assisted in situ surgery (LASIK), photorefractive keratectomy (PRK), laser-assisted sub-epithelial keratectomy (LASEK), corneal ring segments, corneal cross linking, refractive corneal inlays, and corneal lenticular surgery. 26. The method of claim 25, wherein the keratomileusis surgery is LASIK surgery. 27. The method of claim 24, wherein the composition is administered via drops after the surgery. 28. A pharmaceutical kit, comprising a sealed container containing the pharmaceutical composition of claim 1, and an instruction for use of the composition enclosed with the container. | 1,600 |
1,037 | 15,301,473 | 1,613 | The present invention relates to a formulation sized for injection comprising a biodegradable polyesteramide co-polymer comprising at least a diol of bicyclic-1,4:3,6-dianhydrohexitol and analgesics for use in the treatment of arthritic disorders. More specific the invention relates to a formulation comprising injectable microparticle according to claim 1 wherein the polyesteramide co-polymer further comprises a diacid, a diol different from bicyclic-1,4:3,6-dianhydrohexitol and at least two different amino-acids. The formulation is used to treat pain or inflammation in a patient comprising administering to said patient a therapeutically effective amount of the formulation once or twice a year. | 1. A formulation comprising articles sized for injection comprising a biodegradable polyesteramide co-polymer comprising at least a diol of bicyclic-1,4:3,6-dianhydrohexitol for use in the treatment of arthritic disorders. 2. A formulation according to claim 1 wherein the polyesteramide co-polymer further comprises a diacid, a diol different from bicyclic-1,4:3,6-dianhydrohexitol and at least two different amino-acids. 3. A formulation according to claim 1 wherein the polyesteramide co-polymer comprises structural formula (I)
wherein
m varies from 0.01-0.99; p varies from 0.99-0.01; and q varies from 0.99-0.01;
n varies from 5-100
R1 is independently selected from the group consisting of (C2-C20)alkylene, (C2-C20)alkenylene and combinations thereof;
R3 and R4 in a single backbone unit m or p, respectively, are independently selected from the group consisting of hydrogen, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C6-C10)aryl (C1-C6)alkyl, —(CH2)SH, —(CH2)2S(CH3), —CH2OH, —CH(OH)CH3, —(CH2)4NH3+, —CH2COOH, —(CH2)COOH, —CH2—CO—NH2, —CH2CH2—CO—NH2, —CH2CH2COOH, CH3—CH2—CH(CH3)—, —(CH3)2—CH—CH2—, H2N—(CH2)4—, phenyl-CH2—,—CH═CH—CH3, HO-p-phenyl-CH2—, (CH3)2—CH—, phenyl-NH—, NH2—(CH2)3—CH2— or NH2.CH═N—CH═C—CH2—
R5 is selected from the group consisting of (C2-C20) alkylene, (C2-C20) alkenylene.
R6 is selected from bicyclic-fragments of 1,4:3,6-dianhydrohexitols of structural formula (II);
R7 is hydrogen, (C6-C10) aryl, (C1-C6) alkyl or a protecting group such as benzyl;
R8 is independently (C1-C20) alkylene or (C2-C20)alkenyl; 4. A formulation according to claim 3 wherein the polyesteramide co-polymer of Formula (I) comprises m+p+q=1, q=0.25, p=0.45 whereby R1 is —(CH2)8—; R3 and R4 in the backbone units m and p is leucine, —R5 is —(CH2)6—, R6 is a bicyclic-fragments of 1,4:3,6-dianhydrohexitols of structural formula (II); R7 is a benzyl group and R8 is —(CH2)4—. 5. A formulation according to claim 1 further comprising an analgesic selected from the group of anti-inflammatory drugs, local anesthetic drugs and opioids or comprising a disease-modifying antirheumatic drug. 6. A formulation according to claim 5 wherein the analgesic is selected from an NSAID COX-2 inhibitor. 7. A formulation according to claim 5 wherein the analgesic is a corticosteroid. 8. A formulation according to claim 7 wherein the corticosteroid is selected from triamcinolone acetonide. 9. A formulation according to claim 1 wherein the articles are selected from the group of microparticles, fibers, tubes or rods. 10. A formulation according to claim 1 wherein the article is an microparticle. 11. A formulation according to claim 9 wherein the size of the microparticle varies from 0.1-1000 micrometer. 12. A formulation according to claim 1 for use in the treatment of arthritis. 13. A formulation according to claim 12 for use in the treatment of osteoarthritis. 14. A formulation according to claim 13 for use in the treatment of osteoarthritis of the knee, hip, spine or shoulder. 15. A formulation according to claim 1 for use in the treatment of arthritic disorders wherein the formulation is administered in 1 or 2 injections per year. 16. A method of treating pain or inflammation in a human or veterinary patient comprising administering to said patient a therapeutically effective amount of the formulation according to claim 1. 17. A method of slowing, arresting or reversing progressive structural tissue damage associated with chronic inflammatory disease in a human or veterinary patient comprising administering to said patient a therapeutically effective amount of the formulation according to claim 1. 18. The method of claim 16 wherein the formulation is administered in 1 or 2 injections per year. 19. The method of claim 16 wherein the human or veterinary patient has osteoarthritis, rheumatoid arthritis, psoriatic arthritis, autoimmune arthritis, septic arthritis or synovitis. | The present invention relates to a formulation sized for injection comprising a biodegradable polyesteramide co-polymer comprising at least a diol of bicyclic-1,4:3,6-dianhydrohexitol and analgesics for use in the treatment of arthritic disorders. More specific the invention relates to a formulation comprising injectable microparticle according to claim 1 wherein the polyesteramide co-polymer further comprises a diacid, a diol different from bicyclic-1,4:3,6-dianhydrohexitol and at least two different amino-acids. The formulation is used to treat pain or inflammation in a patient comprising administering to said patient a therapeutically effective amount of the formulation once or twice a year.1. A formulation comprising articles sized for injection comprising a biodegradable polyesteramide co-polymer comprising at least a diol of bicyclic-1,4:3,6-dianhydrohexitol for use in the treatment of arthritic disorders. 2. A formulation according to claim 1 wherein the polyesteramide co-polymer further comprises a diacid, a diol different from bicyclic-1,4:3,6-dianhydrohexitol and at least two different amino-acids. 3. A formulation according to claim 1 wherein the polyesteramide co-polymer comprises structural formula (I)
wherein
m varies from 0.01-0.99; p varies from 0.99-0.01; and q varies from 0.99-0.01;
n varies from 5-100
R1 is independently selected from the group consisting of (C2-C20)alkylene, (C2-C20)alkenylene and combinations thereof;
R3 and R4 in a single backbone unit m or p, respectively, are independently selected from the group consisting of hydrogen, (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C6-C10)aryl (C1-C6)alkyl, —(CH2)SH, —(CH2)2S(CH3), —CH2OH, —CH(OH)CH3, —(CH2)4NH3+, —CH2COOH, —(CH2)COOH, —CH2—CO—NH2, —CH2CH2—CO—NH2, —CH2CH2COOH, CH3—CH2—CH(CH3)—, —(CH3)2—CH—CH2—, H2N—(CH2)4—, phenyl-CH2—,—CH═CH—CH3, HO-p-phenyl-CH2—, (CH3)2—CH—, phenyl-NH—, NH2—(CH2)3—CH2— or NH2.CH═N—CH═C—CH2—
R5 is selected from the group consisting of (C2-C20) alkylene, (C2-C20) alkenylene.
R6 is selected from bicyclic-fragments of 1,4:3,6-dianhydrohexitols of structural formula (II);
R7 is hydrogen, (C6-C10) aryl, (C1-C6) alkyl or a protecting group such as benzyl;
R8 is independently (C1-C20) alkylene or (C2-C20)alkenyl; 4. A formulation according to claim 3 wherein the polyesteramide co-polymer of Formula (I) comprises m+p+q=1, q=0.25, p=0.45 whereby R1 is —(CH2)8—; R3 and R4 in the backbone units m and p is leucine, —R5 is —(CH2)6—, R6 is a bicyclic-fragments of 1,4:3,6-dianhydrohexitols of structural formula (II); R7 is a benzyl group and R8 is —(CH2)4—. 5. A formulation according to claim 1 further comprising an analgesic selected from the group of anti-inflammatory drugs, local anesthetic drugs and opioids or comprising a disease-modifying antirheumatic drug. 6. A formulation according to claim 5 wherein the analgesic is selected from an NSAID COX-2 inhibitor. 7. A formulation according to claim 5 wherein the analgesic is a corticosteroid. 8. A formulation according to claim 7 wherein the corticosteroid is selected from triamcinolone acetonide. 9. A formulation according to claim 1 wherein the articles are selected from the group of microparticles, fibers, tubes or rods. 10. A formulation according to claim 1 wherein the article is an microparticle. 11. A formulation according to claim 9 wherein the size of the microparticle varies from 0.1-1000 micrometer. 12. A formulation according to claim 1 for use in the treatment of arthritis. 13. A formulation according to claim 12 for use in the treatment of osteoarthritis. 14. A formulation according to claim 13 for use in the treatment of osteoarthritis of the knee, hip, spine or shoulder. 15. A formulation according to claim 1 for use in the treatment of arthritic disorders wherein the formulation is administered in 1 or 2 injections per year. 16. A method of treating pain or inflammation in a human or veterinary patient comprising administering to said patient a therapeutically effective amount of the formulation according to claim 1. 17. A method of slowing, arresting or reversing progressive structural tissue damage associated with chronic inflammatory disease in a human or veterinary patient comprising administering to said patient a therapeutically effective amount of the formulation according to claim 1. 18. The method of claim 16 wherein the formulation is administered in 1 or 2 injections per year. 19. The method of claim 16 wherein the human or veterinary patient has osteoarthritis, rheumatoid arthritis, psoriatic arthritis, autoimmune arthritis, septic arthritis or synovitis. | 1,600 |
1,038 | 15,068,083 | 1,633 | The present disclosure provides certain silk-fibroin compositions with particular characteristics and/or properties. In some embodiments, the disclosure provides low molecular weight compositions. In some embodiments, the disclosure provides silk fibroin compositions that comprise an active (e.g., a biological) agent or component. In some embodiments, the disclosure provides low molecular weight silk fibroin compositions that comprise an active (e.g., a biological) agent or component. In some embodiments, an active agent is stabilized in a silk composition, e.g., for a period of time and/or against certain conditions or events. In some embodiments, a component present in a silk fibroin composition may be subject to analysis and/or characterization. In some embodiments, a component present in a silk fibroin composition may be recovered from the composition. | 1.-81. (canceled) 82. A composition comprising
a population of silk fibroin fragments; wherein no more than 15% of the total weight of silk fibroin fragments in the population has a molecular weight exceeding 200 kDa, and wherein at least 50% by weight of the silk fibroin fragments have a molecular weight within a range of about 3.5 to about 120 kDa. 83. The composition of claim 82, further comprising at least one additive. 84. The composition of claim 83, wherein the additive is present in a ratio greater than 1:1 as compared to the silk fibroin. 85. The composition of claim 83, wherein the additive comprises at least 5% wt of the composition. 86. The composition of claim 82, wherein the composition comprises at between about 0.1% to 90% silk fibroin by weight. 87. The composition of claim 82, wherein at least 50% by weight of the silk fibroin fragments have a molecular weight within a range of about 19 kDa to 50 kDa. 88. The composition of claim 83, wherein the composition is characterized in that the at least one additive is shelf stable for at least 1 month. 89. The composition of claim 83, wherein the composition is characterized in that at least one property of the at least one additive is stabilized for at least one week. 90. The composition of claim 82, wherein the composition has the form of a gel or hydrogel, powder, particle, foam, lyophilized article, and combinations thereof. 91. The composition of claim 82, wherein the silk fibroin fragments have a continuous molecular weight distribution within the range. 92. The composition of claim 82, wherein the silk fibroin fragments have a discrete molecular weight distribution within the range. 93. The composition of claim 83, wherein the at least one additive is selected from the group consisting of small organic or inorganic molecules, saccharides, oligosaccharides, polysaccharides, polymers, proteins, peptides, peptide analogs, peptide derivatives, peptidomimetics, nucleic acids, nucleic acid analogs, and combinations thereof. 94. The composition of claim 83, wherein the additive is or comprises an immunogen, cell attachment mediator, stabilizer, biocompatible polymer, growth factor, anti-inflammatory agent, antimicrobial agent, a vaccine, biological sample, or combination thereof. 95. The composition of claim 94, wherein the cell attachment mediator is selected from the group consisting of collagen, elastin, fibronectin, vitronectin, laminin, proteoglycans, a peptide comprising a known integrin binding domain, and combinations thereof. 96. The composition of claim 94, wherein the biological sample is selected from the group consisting of tissue, blood, plasma, serum, urine, cerebrospinal fluid, saliva, hormones, growth factors, cytokines, antibodies, and any combination thereof. 97. The composition of claim 82, further comprising at least one additional material for soft tissue augmentation. 98. The composition of claim 97, wherein the at least one additional material for soft tissue augmentation is or comprises at least one of poly(methyl methacrylate), microspheres, hydroxyapatite, poly(L-lactic acid), collagen, elastin, a glycosaminoglycan, hyaluronic acid, BOTOX®, DYSPORT®, COSMODERM®, EVOLENCE®, RADIESSE®, RESTYLANE®, JUVEDERM®, SCULPTRA®, PERLANE®, CAPTIQUE®, and combinations thereof. 99. The composition of claim 82, wherein the composition has a solubility of at least 5% in an aqueous solution. 100. The composition of claim 82, wherein the composition can be resolubilized in an aqueous solution at room temperature. 101. The composition of claim 82, wherein the composition has been subjected to at least one purification step. 102. The composition of claim 101, wherein the at least one purification step is or comprises filtration. | The present disclosure provides certain silk-fibroin compositions with particular characteristics and/or properties. In some embodiments, the disclosure provides low molecular weight compositions. In some embodiments, the disclosure provides silk fibroin compositions that comprise an active (e.g., a biological) agent or component. In some embodiments, the disclosure provides low molecular weight silk fibroin compositions that comprise an active (e.g., a biological) agent or component. In some embodiments, an active agent is stabilized in a silk composition, e.g., for a period of time and/or against certain conditions or events. In some embodiments, a component present in a silk fibroin composition may be subject to analysis and/or characterization. In some embodiments, a component present in a silk fibroin composition may be recovered from the composition.1.-81. (canceled) 82. A composition comprising
a population of silk fibroin fragments; wherein no more than 15% of the total weight of silk fibroin fragments in the population has a molecular weight exceeding 200 kDa, and wherein at least 50% by weight of the silk fibroin fragments have a molecular weight within a range of about 3.5 to about 120 kDa. 83. The composition of claim 82, further comprising at least one additive. 84. The composition of claim 83, wherein the additive is present in a ratio greater than 1:1 as compared to the silk fibroin. 85. The composition of claim 83, wherein the additive comprises at least 5% wt of the composition. 86. The composition of claim 82, wherein the composition comprises at between about 0.1% to 90% silk fibroin by weight. 87. The composition of claim 82, wherein at least 50% by weight of the silk fibroin fragments have a molecular weight within a range of about 19 kDa to 50 kDa. 88. The composition of claim 83, wherein the composition is characterized in that the at least one additive is shelf stable for at least 1 month. 89. The composition of claim 83, wherein the composition is characterized in that at least one property of the at least one additive is stabilized for at least one week. 90. The composition of claim 82, wherein the composition has the form of a gel or hydrogel, powder, particle, foam, lyophilized article, and combinations thereof. 91. The composition of claim 82, wherein the silk fibroin fragments have a continuous molecular weight distribution within the range. 92. The composition of claim 82, wherein the silk fibroin fragments have a discrete molecular weight distribution within the range. 93. The composition of claim 83, wherein the at least one additive is selected from the group consisting of small organic or inorganic molecules, saccharides, oligosaccharides, polysaccharides, polymers, proteins, peptides, peptide analogs, peptide derivatives, peptidomimetics, nucleic acids, nucleic acid analogs, and combinations thereof. 94. The composition of claim 83, wherein the additive is or comprises an immunogen, cell attachment mediator, stabilizer, biocompatible polymer, growth factor, anti-inflammatory agent, antimicrobial agent, a vaccine, biological sample, or combination thereof. 95. The composition of claim 94, wherein the cell attachment mediator is selected from the group consisting of collagen, elastin, fibronectin, vitronectin, laminin, proteoglycans, a peptide comprising a known integrin binding domain, and combinations thereof. 96. The composition of claim 94, wherein the biological sample is selected from the group consisting of tissue, blood, plasma, serum, urine, cerebrospinal fluid, saliva, hormones, growth factors, cytokines, antibodies, and any combination thereof. 97. The composition of claim 82, further comprising at least one additional material for soft tissue augmentation. 98. The composition of claim 97, wherein the at least one additional material for soft tissue augmentation is or comprises at least one of poly(methyl methacrylate), microspheres, hydroxyapatite, poly(L-lactic acid), collagen, elastin, a glycosaminoglycan, hyaluronic acid, BOTOX®, DYSPORT®, COSMODERM®, EVOLENCE®, RADIESSE®, RESTYLANE®, JUVEDERM®, SCULPTRA®, PERLANE®, CAPTIQUE®, and combinations thereof. 99. The composition of claim 82, wherein the composition has a solubility of at least 5% in an aqueous solution. 100. The composition of claim 82, wherein the composition can be resolubilized in an aqueous solution at room temperature. 101. The composition of claim 82, wherein the composition has been subjected to at least one purification step. 102. The composition of claim 101, wherein the at least one purification step is or comprises filtration. | 1,600 |
1,039 | 15,213,055 | 1,613 | This invention discloses drug-delivery compositions, methods of making prodrugs, and methods of drug delivery using a self-assembled gelator. The backbone of the gelator can contain a drug or prodrug, such as acetaminophen or salicin. Additional drugs or agents can be encapsulated in the gelator. Enzymatic or hydrolytic cleavage can be used to release the drugs. | 1-166. (canceled) 167. A drug delivery gel composition comprising one or more drugs for delivery, the composition comprising
a gel comprising a plurality of self-assembling amphiphilic drug-derived gelators, the self-assembling amphiphilic drug-derived gelators comprising a headgroup conjugated via a degradable linkage to a tail group comprising a hydrophilic moiety or hydrophobic moiety, the gelator comprising a drug or prodrug moiety; wherein the self-assembling amphiphilic drug-derived gelators self-assemble and precipitate when exposed to a change in pH, temperature, solvent, concentration, or combination thereof to form lamellar fiber structures, the lamellar fiber structures controlling release of the drug or prodrug moiety and any additional drugs. 168. The gel of claim 167, wherein the gel is a hydrogel. 169. The gel of claim 167, wherein the gel is an organogel. 170. The gel of claim 167, wherein the degradable linkage is selected from the group consisting of an ester, amide, carbamate, disulfide, and anhydride. 171. The gel of claim 170, wherein the degradable linkage is cleavable by enzyme-mediated degradation or by hydrolytic cleavage to release the headgroup from the tail group, thereby releasing the drug or prodrug moiety from the backbone of the self-assembled gelators and from the gel. 172. The gel of claim 167, wherein the headgroup is hydrophilic and the tail group is hydrophobic. 173. The gel of claim 167, wherein the headgroup is hydrophobic and the tail group is hydrophilic. 174. The gel of claim 167, wherein the one or more drugs is selected from the group consisting of chemotherapeutic drugs, antibiotic drugs, analgesic, anti-pyretic, anti-inflammatory drugs, and combinations thereof. 175. The gel of claim 174, wherein the drug moiety of the gelator comprises an anti-inflammatory drug moiety selected from the group consisting of acetaminophen, salicin, indomethacin, and prodrugs and derivatives thereof. 176. The gel of claim 167, wherein the fiber structures have a thickness of about 50 to 400 nm. 177. The gel of claim 176, wherein the fiber structures have a thickness of about 100 to 200 nm. 178. The gel of claim 167, comprising one or more drugs encapsulated in the gel. 179. The gel of claim 177, comprising two or more drugs. 180. The gel of claim 177, wherein the degradable linkage is cleavable by enzyme-mediated degradation or by hydrolytic cleavage; and
the one or more drugs are released upon exposure of the gel to an enzyme and/or upon hydrolysis of the gel. 181. The gel of claim 180, wherein the release of the one or more drugs is sustained release, not burst release. 182. A drug delivery gel composition comprising one or more drugs for delivery, the composition comprising
a gel comprising a plurality of self-assembled amphiphilic drug-derived gelators, the self-assembled amphiphilic drug-derived gelators comprising a headgroup conjugated via a degradable linkage to a tail group comprising a hydrophilic moiety or hydrophobic moiety, the gelator comprising a drug or prodrug moiety; wherein the self-assembled amphiphilic drug-derived gelators are formed into lamellar fiber structures when exposed to a change in pH, temperature, solvent, concentration, or combination thereof, the lamellar fiber structures controlling release of the drug or prodrug moiety and any additional drugs. 183. The gel composition of claim 182, wherein the gel is a hydrogel. 184. The gel composition of claim 182, wherein the gel is an organogel. 185. The gel composition of claim 182, wherein the degradable linkage is selected from the group consisting of an ester, amide, carbamate, disulfide, and anhydride. 186. The gel composition of claim 185, wherein the degradable linkage is cleavable by enzyme-mediated degradation or by hydrolytic cleavage to release the headgroup from the tail group; thereby releasing the drug or prodrug moiety from the backbone of the self-assembled gelators and from the gel. 187. The gel composition of claim 182, wherein the headgroup is hydrophilic and the tail group is hydrophobic. 188. The gel composition of claim 182, wherein the headgroup is hydrophobic and the tail group is hydrophilic. 189. The gel composition of claim 182, wherein the one or more drugs is selected from the group consisting of chemotherapeutic drugs, antibiotic drugs, analgesic, anti-pyretic, anti-inflammatory drugs, and combinations thereof. 190. The gel composition of claim 189, wherein the drug moiety of the gelator comprises an anti-inflammatory drug moiety selected from the group consisting of acetaminophen, salicin, indomethacin, and prodrugs and derivatives thereof. 191. The gel composition of claim 182, wherein the fiber structures have a thickness of about 50 to 400 nm. 192. The gel composition of claim 191, wherein the fiber structures have a thickness of about 100 to 200 nm. 193. The gel composition of claim 182, comprising one or more drugs encapsulated in the gel. 194. The gel composition of claim 182, comprising two or more drugs. 196. The gel composition of claim 182, wherein the release of the one or more encapsulated agents is a sustained release and not burst release. | This invention discloses drug-delivery compositions, methods of making prodrugs, and methods of drug delivery using a self-assembled gelator. The backbone of the gelator can contain a drug or prodrug, such as acetaminophen or salicin. Additional drugs or agents can be encapsulated in the gelator. Enzymatic or hydrolytic cleavage can be used to release the drugs.1-166. (canceled) 167. A drug delivery gel composition comprising one or more drugs for delivery, the composition comprising
a gel comprising a plurality of self-assembling amphiphilic drug-derived gelators, the self-assembling amphiphilic drug-derived gelators comprising a headgroup conjugated via a degradable linkage to a tail group comprising a hydrophilic moiety or hydrophobic moiety, the gelator comprising a drug or prodrug moiety; wherein the self-assembling amphiphilic drug-derived gelators self-assemble and precipitate when exposed to a change in pH, temperature, solvent, concentration, or combination thereof to form lamellar fiber structures, the lamellar fiber structures controlling release of the drug or prodrug moiety and any additional drugs. 168. The gel of claim 167, wherein the gel is a hydrogel. 169. The gel of claim 167, wherein the gel is an organogel. 170. The gel of claim 167, wherein the degradable linkage is selected from the group consisting of an ester, amide, carbamate, disulfide, and anhydride. 171. The gel of claim 170, wherein the degradable linkage is cleavable by enzyme-mediated degradation or by hydrolytic cleavage to release the headgroup from the tail group, thereby releasing the drug or prodrug moiety from the backbone of the self-assembled gelators and from the gel. 172. The gel of claim 167, wherein the headgroup is hydrophilic and the tail group is hydrophobic. 173. The gel of claim 167, wherein the headgroup is hydrophobic and the tail group is hydrophilic. 174. The gel of claim 167, wherein the one or more drugs is selected from the group consisting of chemotherapeutic drugs, antibiotic drugs, analgesic, anti-pyretic, anti-inflammatory drugs, and combinations thereof. 175. The gel of claim 174, wherein the drug moiety of the gelator comprises an anti-inflammatory drug moiety selected from the group consisting of acetaminophen, salicin, indomethacin, and prodrugs and derivatives thereof. 176. The gel of claim 167, wherein the fiber structures have a thickness of about 50 to 400 nm. 177. The gel of claim 176, wherein the fiber structures have a thickness of about 100 to 200 nm. 178. The gel of claim 167, comprising one or more drugs encapsulated in the gel. 179. The gel of claim 177, comprising two or more drugs. 180. The gel of claim 177, wherein the degradable linkage is cleavable by enzyme-mediated degradation or by hydrolytic cleavage; and
the one or more drugs are released upon exposure of the gel to an enzyme and/or upon hydrolysis of the gel. 181. The gel of claim 180, wherein the release of the one or more drugs is sustained release, not burst release. 182. A drug delivery gel composition comprising one or more drugs for delivery, the composition comprising
a gel comprising a plurality of self-assembled amphiphilic drug-derived gelators, the self-assembled amphiphilic drug-derived gelators comprising a headgroup conjugated via a degradable linkage to a tail group comprising a hydrophilic moiety or hydrophobic moiety, the gelator comprising a drug or prodrug moiety; wherein the self-assembled amphiphilic drug-derived gelators are formed into lamellar fiber structures when exposed to a change in pH, temperature, solvent, concentration, or combination thereof, the lamellar fiber structures controlling release of the drug or prodrug moiety and any additional drugs. 183. The gel composition of claim 182, wherein the gel is a hydrogel. 184. The gel composition of claim 182, wherein the gel is an organogel. 185. The gel composition of claim 182, wherein the degradable linkage is selected from the group consisting of an ester, amide, carbamate, disulfide, and anhydride. 186. The gel composition of claim 185, wherein the degradable linkage is cleavable by enzyme-mediated degradation or by hydrolytic cleavage to release the headgroup from the tail group; thereby releasing the drug or prodrug moiety from the backbone of the self-assembled gelators and from the gel. 187. The gel composition of claim 182, wherein the headgroup is hydrophilic and the tail group is hydrophobic. 188. The gel composition of claim 182, wherein the headgroup is hydrophobic and the tail group is hydrophilic. 189. The gel composition of claim 182, wherein the one or more drugs is selected from the group consisting of chemotherapeutic drugs, antibiotic drugs, analgesic, anti-pyretic, anti-inflammatory drugs, and combinations thereof. 190. The gel composition of claim 189, wherein the drug moiety of the gelator comprises an anti-inflammatory drug moiety selected from the group consisting of acetaminophen, salicin, indomethacin, and prodrugs and derivatives thereof. 191. The gel composition of claim 182, wherein the fiber structures have a thickness of about 50 to 400 nm. 192. The gel composition of claim 191, wherein the fiber structures have a thickness of about 100 to 200 nm. 193. The gel composition of claim 182, comprising one or more drugs encapsulated in the gel. 194. The gel composition of claim 182, comprising two or more drugs. 196. The gel composition of claim 182, wherein the release of the one or more encapsulated agents is a sustained release and not burst release. | 1,600 |
1,040 | 16,128,053 | 1,627 | Disclosed herein are safer formulations of resiniferatoxin (RTX) for intrathecal, intraganglionic intraarticular and pericardial administration. More specifically, there is disclosed alcohol-free formulations of RTX comprising a solubilizing component, a monosaccharide or sugar alcohol, a saline buffer, and RTX, and having narrow ranges for pH range and specific gravity. | 1. A non-alcoholic formulation of RTX comprising from about 10 μg/mL to about 200 μg/mL RTX solubilized in a solubilizing agent, a monosaccharide or sugar alcohol, and a buffer solution, wherein the formulation has a pH from about 6.5 to about 7.5. 2. The non-alcoholic formulation of RTX of claim 1, wherein the solubilizing agent is selected from the group consisting of PEG, polysorbate and cyclodextrin, or combinations thereof. 3. The non-alcoholic formulation of RTX of claim 1, wherein the formulation comprises from about 25-50 μg/mL RTX. 4. The non-alcoholic formulation of RTX of claim 1, wherein the monosaccharide or sugar alcohol is selected from the group consisting of dextrose and mannitol, or combinations thereof. 5. The non-alcoholic formulation of RTX of claim 1, wherein the saline buffer is selected from the group consisting of a phosphate buffer, an acetate buffer, and a citrate buffer, or combinations thereof. 6. The non-alcoholic formulation of RTX of claim 1, further comprising an antioxidant. 7. The non-alcoholic formulation of RTX of claim 6, wherein the antioxidant is selected from the group consisting of ascorbic acid, citric acid, potassium bisulfate, sodium bisulfate acetone sodium bisulfate, monothioglycerol, potassium metabisulfite, and sodium metabisulfite, or combinations thereof. 8. The non-alcoholic formulation of RTX of claim 2, wherein the solubilizing agent is selected from the group consisting of PEG (0-40%) , polysorbate (0-5%) and cyclodextrin (0-5%), or combinations thereof. 9. The non-alcoholic formulation of RTX of claim 1, comprising from about 10 μg/mL to about 200 μg/mL RTX solubilized in polysorbate 80, dextrose, and a phosphate buffer solution, wherein the formulation has a pH from about 6.5 to about 7.5. 10. The non-alcoholic formulation of RTX of claim 9, comprising 200 μg/mL RTX solubilized in 0.03% v/v polysorbate 80, 0.05% w/v dextrose, and 30 mM phosphate buffer solution, wherein the formulation has a pH of about 7.2. | Disclosed herein are safer formulations of resiniferatoxin (RTX) for intrathecal, intraganglionic intraarticular and pericardial administration. More specifically, there is disclosed alcohol-free formulations of RTX comprising a solubilizing component, a monosaccharide or sugar alcohol, a saline buffer, and RTX, and having narrow ranges for pH range and specific gravity.1. A non-alcoholic formulation of RTX comprising from about 10 μg/mL to about 200 μg/mL RTX solubilized in a solubilizing agent, a monosaccharide or sugar alcohol, and a buffer solution, wherein the formulation has a pH from about 6.5 to about 7.5. 2. The non-alcoholic formulation of RTX of claim 1, wherein the solubilizing agent is selected from the group consisting of PEG, polysorbate and cyclodextrin, or combinations thereof. 3. The non-alcoholic formulation of RTX of claim 1, wherein the formulation comprises from about 25-50 μg/mL RTX. 4. The non-alcoholic formulation of RTX of claim 1, wherein the monosaccharide or sugar alcohol is selected from the group consisting of dextrose and mannitol, or combinations thereof. 5. The non-alcoholic formulation of RTX of claim 1, wherein the saline buffer is selected from the group consisting of a phosphate buffer, an acetate buffer, and a citrate buffer, or combinations thereof. 6. The non-alcoholic formulation of RTX of claim 1, further comprising an antioxidant. 7. The non-alcoholic formulation of RTX of claim 6, wherein the antioxidant is selected from the group consisting of ascorbic acid, citric acid, potassium bisulfate, sodium bisulfate acetone sodium bisulfate, monothioglycerol, potassium metabisulfite, and sodium metabisulfite, or combinations thereof. 8. The non-alcoholic formulation of RTX of claim 2, wherein the solubilizing agent is selected from the group consisting of PEG (0-40%) , polysorbate (0-5%) and cyclodextrin (0-5%), or combinations thereof. 9. The non-alcoholic formulation of RTX of claim 1, comprising from about 10 μg/mL to about 200 μg/mL RTX solubilized in polysorbate 80, dextrose, and a phosphate buffer solution, wherein the formulation has a pH from about 6.5 to about 7.5. 10. The non-alcoholic formulation of RTX of claim 9, comprising 200 μg/mL RTX solubilized in 0.03% v/v polysorbate 80, 0.05% w/v dextrose, and 30 mM phosphate buffer solution, wherein the formulation has a pH of about 7.2. | 1,600 |
1,041 | 15,767,116 | 1,618 | Compositions and methods for administration of local anesthetics that are delivered by a single injection and enable repeated on-demand or high influx analgesia over extended periods have been developed. Pharmaceutical compositions including an effective amount of one or more sodium channel blockers including site 1 sodium channel blockers, optionally one or more alpha-2-adrenergic agonists, which are optionally encapsulated in liposomes, particles or microbubbles, and one or more triggerable elements are provided. The triggerable elements allow delivery of the encapsulated anesthetic drugs when an appropriate triggering stimuli are applied. Exemplary triggering agents or stimuli include near-infrared irradiation, UV- and visible light, ultrasound and magnetic field. In one embodiment, ultrasound is used to trigger a burst of microbubbles to enhance penetration of local anesthetic. | 1. A pharmaceutical composition comprising
triggerable liposomes, particles or microbubbles encapsulating at least one site I sodium channel blocker, wherein the site I sodium channel blocker is released upon exposing to a triggering agent in an amount effective to produce anesthesia. 2. The pharmaceutical composition of claim 1, wherein the triggering agent is selected from the group consisting of near-infrared irradiation, ultraviolet and visible light, ultrasound and magnetic field. 3. The pharmaceutical composition of claim 1, wherein the liposomes or particles contain one or more triggerable elements. 4. The pharmaceutical composition of claim 3, comprising liposomes, wherein the triggerable elements are any material that disrupts membrane bilayer to release the encapsulated content in response to any of the triggering agents. 5. The pharmaceutical composition of claim 1, further comprising one or more alpha-2-adrenergic agonist, local anesthetic or vasoconstrictor encapsulated in liposomes or particles. 6. The pharmaceutical composition of claim 4, wherein the triggerable element is a gold nanoparticle selected from the group consisting of gold nanorods, gold nanoshells, gold nanostars and gold nanocages. 7. The pharmaceutical composition of claim 6, wherein the gold nanoparticle is gold nanorods. 8. The pharmaceutical composition of claim 7, wherein the gold nanorods have an aspect ratio ranging from 1.5 to 10 for which the surface plasmon absorption maxima are between 600 and 1300 nm. 9. The pharmaceutical composition of claim 7, wherein the gold nanorods have a concentration of between 0.001 and 1 wt %. 10. The pharmaceutical composition of claim 3, wherein the triggerable element is a photosensitizer. 11. The pharmaceutical composition of claim 10, wherein the photosensitizer comprises 1,4,8,11,15,18, 22,25-octabutoxyphthalocyaninato-palladium(II), PdPc(OBu)8. 12. The pharmaceutical composition of claim 11 comprising liposomes formed of unsaturated lipids such as Egg PC (L-α-phosphatidylcholine), DLPC (1,2-dilinoleoyl-sn-glycero-3-phosphocholine), or other lipids with unsaturated bonds combined with DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DSPG (1,2-dioctadecanoyl-sn-glycero-3-phospho-(1′-rac-glycerol)) and cholesterol to form the liposomes. 13. The pharmaceutical composition of claim 3, wherein the triggerable element comprises a sonosensitizer. 14. The pharmaceutical composition of claim 13, wherein the sonosensitizer comprises protoporphyrin IX. 15. The pharmaceutical composition of claim 1, comprising liposomes having a phase transition temperature that is between 33 and 43° C. 16. The pharmaceutical composition of claim 15, wherein the liposome comprises 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2¬dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG), cholesterol and thiolated PEG-DSPE (HS-PEG-DSPE). 17. The pharmaceutical composition of claim 16, wherein the total lipid comprises a molar ratio of 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) to 1,2¬dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG) to cholesterol to thiolated PEG-DSPE (HS-PEG-DSPE) of about 6:2:3:0.2. 18. The pharmaceutical composition of claim 1 comprising microbubbles which enhance penetration of the anesthetic to the nerves where anesthesia is desired. 19. The pharmaceutical composition of claim 18 wherein ultrasound is used to further enhanced penetration of the microbubbles and anesthetics. 20. The pharmaceutical composition of claim 1, wherein the site 1 sodium channel blocker is selected from the group consisting of tetrodotoxin; saxitoxin; decarbamoyl saxitoxin; Neosaxitoxin; gonyautoxins; and conotoxins. 21. The pharmaceutical composition of claim 20, wherein one site 1 sodium channel blocker is tetrodotoxin. 22. The pharmaceutical composition of claim 5 comprising an alpha-2-adrenergic agonist is selected from the group consisting of xylazine, flutonidine, moxonidine, tramazoline, tolonidine, piclonidine, tiamenidine, clonidine and dexmedetomidine. 23. The pharmaceutical composition of claim 22, wherein the alpha-2-adrenergic agonist is dexmedetomidine. 24. The pharmaceutical composition of claim 1 comprising polymeric particles. 25. The pharmaceutical composition of claim 1, further comprising a pharmaceutically acceptable carrier. 26. The pharmaceutical composition of claim 1 in an amount effective for the treatment or prevention of pain at or near the site of administration in an awake, sedated or anesthetized human. 27. A method for the treatment or prevention of pain via controlling the frequency, wavelength, intensity and duration of the release of nerve block in the absence of local toxicity, comprising
a) administering to the subject at or near the site of the pain an effective amount of the pharmaceutical composition of claim 1, b) applying a triggering agent such light, ultrasound and magnetic field at the site of treatment on the patient for a sufficient release of the encapsulated site I sodium channel blocker and any other agents. 28. The method of claim 27 further comprising removing the triggering source to prevent further release once the level of pain relief is achieved. 29. The method of claim 27 comprising repeating triggering release of the site I sodium channel blocker to provide additional anesthesia. 30. The method of claim 27, wherein the light is near-infrared radiation ranging between 600 and 1300 nm. 31. The method of claim 27, wherein the application of the triggering agent lasts up to 30 min for each trigger cycle. 32. The method of claim 27, wherein the triggerable drug release can be repeated three or more times when further dosage is desired. 33. The method of claim 27 wherein repeatable and adjustable peripheral nerve blockade lasts for up to five or more days following a single application. 34. The method of claim 27 for providing nerve blockade comprising administering to a site in an individual a composition comprising microbubbles and at least one site I sodium channel blocker in an effective amount, and applying ultrasound at the site, to provide a prolonged nerve blockade in the absence of local myotoxicity. 35. The method of claim 34, wherein the site 1 sodium channel blocker is selected from the group consisting of tetrodotoxin (TTX), saxitoxin (STX), decarbamoyl saxitoxin, neosaxitoxin, and the gonyautoxins. 36. The method of claim 35, wherein the site 1 sodium channel blocker is TTX. 37. The method of claim 34, wherein the microbubbles are formed from one or more lipids selected from the group consisting of 1,2-distearoyl-sn-glycero-3 phosphocholine (DSPC), 1,2 distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), and derivatives thereof. 38. The method of claim 35, wherein the derivatives include polyethylene glycol (PEG)-conjugated DSPE and PEG-conjugated DSPC. 39. The method of claim 35, wherein the ultrasound is administered at a frequency between 100 kHz and 2.5 MHz, preferably between 1 MHz and 2.5 MHz. 40. The method of claim 35, wherein the ultrasound is administered at an intensity less than 1 W/cm2, less than 0.1 W/cm2, and most preferably or less than about 0.02 W/cm2. 41. The method of claim 35, wherein the nerve blockade has a prolonged duration of sensory nerve block and motor nerve block. | Compositions and methods for administration of local anesthetics that are delivered by a single injection and enable repeated on-demand or high influx analgesia over extended periods have been developed. Pharmaceutical compositions including an effective amount of one or more sodium channel blockers including site 1 sodium channel blockers, optionally one or more alpha-2-adrenergic agonists, which are optionally encapsulated in liposomes, particles or microbubbles, and one or more triggerable elements are provided. The triggerable elements allow delivery of the encapsulated anesthetic drugs when an appropriate triggering stimuli are applied. Exemplary triggering agents or stimuli include near-infrared irradiation, UV- and visible light, ultrasound and magnetic field. In one embodiment, ultrasound is used to trigger a burst of microbubbles to enhance penetration of local anesthetic.1. A pharmaceutical composition comprising
triggerable liposomes, particles or microbubbles encapsulating at least one site I sodium channel blocker, wherein the site I sodium channel blocker is released upon exposing to a triggering agent in an amount effective to produce anesthesia. 2. The pharmaceutical composition of claim 1, wherein the triggering agent is selected from the group consisting of near-infrared irradiation, ultraviolet and visible light, ultrasound and magnetic field. 3. The pharmaceutical composition of claim 1, wherein the liposomes or particles contain one or more triggerable elements. 4. The pharmaceutical composition of claim 3, comprising liposomes, wherein the triggerable elements are any material that disrupts membrane bilayer to release the encapsulated content in response to any of the triggering agents. 5. The pharmaceutical composition of claim 1, further comprising one or more alpha-2-adrenergic agonist, local anesthetic or vasoconstrictor encapsulated in liposomes or particles. 6. The pharmaceutical composition of claim 4, wherein the triggerable element is a gold nanoparticle selected from the group consisting of gold nanorods, gold nanoshells, gold nanostars and gold nanocages. 7. The pharmaceutical composition of claim 6, wherein the gold nanoparticle is gold nanorods. 8. The pharmaceutical composition of claim 7, wherein the gold nanorods have an aspect ratio ranging from 1.5 to 10 for which the surface plasmon absorption maxima are between 600 and 1300 nm. 9. The pharmaceutical composition of claim 7, wherein the gold nanorods have a concentration of between 0.001 and 1 wt %. 10. The pharmaceutical composition of claim 3, wherein the triggerable element is a photosensitizer. 11. The pharmaceutical composition of claim 10, wherein the photosensitizer comprises 1,4,8,11,15,18, 22,25-octabutoxyphthalocyaninato-palladium(II), PdPc(OBu)8. 12. The pharmaceutical composition of claim 11 comprising liposomes formed of unsaturated lipids such as Egg PC (L-α-phosphatidylcholine), DLPC (1,2-dilinoleoyl-sn-glycero-3-phosphocholine), or other lipids with unsaturated bonds combined with DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DSPG (1,2-dioctadecanoyl-sn-glycero-3-phospho-(1′-rac-glycerol)) and cholesterol to form the liposomes. 13. The pharmaceutical composition of claim 3, wherein the triggerable element comprises a sonosensitizer. 14. The pharmaceutical composition of claim 13, wherein the sonosensitizer comprises protoporphyrin IX. 15. The pharmaceutical composition of claim 1, comprising liposomes having a phase transition temperature that is between 33 and 43° C. 16. The pharmaceutical composition of claim 15, wherein the liposome comprises 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2¬dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG), cholesterol and thiolated PEG-DSPE (HS-PEG-DSPE). 17. The pharmaceutical composition of claim 16, wherein the total lipid comprises a molar ratio of 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) to 1,2¬dipalmitoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (DPPG) to cholesterol to thiolated PEG-DSPE (HS-PEG-DSPE) of about 6:2:3:0.2. 18. The pharmaceutical composition of claim 1 comprising microbubbles which enhance penetration of the anesthetic to the nerves where anesthesia is desired. 19. The pharmaceutical composition of claim 18 wherein ultrasound is used to further enhanced penetration of the microbubbles and anesthetics. 20. The pharmaceutical composition of claim 1, wherein the site 1 sodium channel blocker is selected from the group consisting of tetrodotoxin; saxitoxin; decarbamoyl saxitoxin; Neosaxitoxin; gonyautoxins; and conotoxins. 21. The pharmaceutical composition of claim 20, wherein one site 1 sodium channel blocker is tetrodotoxin. 22. The pharmaceutical composition of claim 5 comprising an alpha-2-adrenergic agonist is selected from the group consisting of xylazine, flutonidine, moxonidine, tramazoline, tolonidine, piclonidine, tiamenidine, clonidine and dexmedetomidine. 23. The pharmaceutical composition of claim 22, wherein the alpha-2-adrenergic agonist is dexmedetomidine. 24. The pharmaceutical composition of claim 1 comprising polymeric particles. 25. The pharmaceutical composition of claim 1, further comprising a pharmaceutically acceptable carrier. 26. The pharmaceutical composition of claim 1 in an amount effective for the treatment or prevention of pain at or near the site of administration in an awake, sedated or anesthetized human. 27. A method for the treatment or prevention of pain via controlling the frequency, wavelength, intensity and duration of the release of nerve block in the absence of local toxicity, comprising
a) administering to the subject at or near the site of the pain an effective amount of the pharmaceutical composition of claim 1, b) applying a triggering agent such light, ultrasound and magnetic field at the site of treatment on the patient for a sufficient release of the encapsulated site I sodium channel blocker and any other agents. 28. The method of claim 27 further comprising removing the triggering source to prevent further release once the level of pain relief is achieved. 29. The method of claim 27 comprising repeating triggering release of the site I sodium channel blocker to provide additional anesthesia. 30. The method of claim 27, wherein the light is near-infrared radiation ranging between 600 and 1300 nm. 31. The method of claim 27, wherein the application of the triggering agent lasts up to 30 min for each trigger cycle. 32. The method of claim 27, wherein the triggerable drug release can be repeated three or more times when further dosage is desired. 33. The method of claim 27 wherein repeatable and adjustable peripheral nerve blockade lasts for up to five or more days following a single application. 34. The method of claim 27 for providing nerve blockade comprising administering to a site in an individual a composition comprising microbubbles and at least one site I sodium channel blocker in an effective amount, and applying ultrasound at the site, to provide a prolonged nerve blockade in the absence of local myotoxicity. 35. The method of claim 34, wherein the site 1 sodium channel blocker is selected from the group consisting of tetrodotoxin (TTX), saxitoxin (STX), decarbamoyl saxitoxin, neosaxitoxin, and the gonyautoxins. 36. The method of claim 35, wherein the site 1 sodium channel blocker is TTX. 37. The method of claim 34, wherein the microbubbles are formed from one or more lipids selected from the group consisting of 1,2-distearoyl-sn-glycero-3 phosphocholine (DSPC), 1,2 distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), and derivatives thereof. 38. The method of claim 35, wherein the derivatives include polyethylene glycol (PEG)-conjugated DSPE and PEG-conjugated DSPC. 39. The method of claim 35, wherein the ultrasound is administered at a frequency between 100 kHz and 2.5 MHz, preferably between 1 MHz and 2.5 MHz. 40. The method of claim 35, wherein the ultrasound is administered at an intensity less than 1 W/cm2, less than 0.1 W/cm2, and most preferably or less than about 0.02 W/cm2. 41. The method of claim 35, wherein the nerve blockade has a prolonged duration of sensory nerve block and motor nerve block. | 1,600 |
1,042 | 15,466,187 | 1,611 | The present invention relates to a Tabletting aid with a low water content and to a process for the preparation thereof. The Tabletting aid composition is a directly compressible composition, the use of which results in improved tablet properties. | 1.-20. (canceled) 21. A directly compressible tabletting composition, comprising 50-85% by weight of anhydrous calcium hydrogenphosphate, 10-40% by weight of mannitol and 5-20% by weight of sorbitol, wherein said composition has a flow angle in the range of 29 to 33.4° and a bulk density in the range of 0.56 to 0.77 g/ml with a tapped density in the range of 0.73 to 0.92 g/ml. 22. The directly compressible tabletting composition according to claim 21, further comprising one or more homogeneously distributed, water-insoluble and/or water-soluble additives. 23. The directly compressible tabletting composition according to claim 21, one or more one or more homogeneously distributed, water-insoluble and/or water-soluble additives selected from: pharmaceutical active compounds, plant extracts, sweeteners, dyes, citric acid, vitamins and trace elements. 24. The directly compressible tabletting composition according to claim 23, which comprises one or more pharmaceutical active compounds which are analgesics. 25. The directly compressible tabletting composition according to claim 23, which comprises one or more sweeteners selected from: acesulfame K, Aspartame®, saccharin, cyclamate, sucralose and neohesperidin DC. 26. The directly compressible tabletting composition according to claim 21, which comprises 60 to 80% by weight of anhydrous calcium hydrogenphosphate, 15 to 25% by weight of mannitol and 7 to 13% by weight of sorbitol. 27. The directly compressible tabletting composition according to claim 21, which comprises 65 to 75% by weight of anhydrous calcium hydrogenphosphate, 17 to 23% by weight of mannitol and 8 to 12% by weight of sorbitol. 28. The directly compressible tabletting composition according to claim 21, wherein the anhydrous calcium hydrogenphosphate, mannitol and sorbitol are co-spray granulated. 29. The directly compressible tabletting composition according to claim 21, wherein the composition has a particle-size distribution of max. 3% by weight of undersized particles having a particle size of <32 μm, max. 5% by weight of oversized particles having a particle size of >500 μm, and 50 to 90% by weight of a particle fraction having particle sizes in the range from 100 to 315 μm. 30. The directly compressible tabletting composition according to claim 21, wherein the composition has a calcium content of 14 to 21% by weight, based on the total amount, and a drying loss of less than 2% by weight. 31. The directly compressible tabletting composition according to claim 21, wherein the composition has a calcium content of 14 to 21% by weight, based on the total amount, and a drying loss of less than 1% by weight. 32. The directly compressible tabletting composition according to claim 21, wherein the composition is in the form of tablets formed by compression with a pressing force of 20 kN, and the tablets have a hardness of >270 N, together with an ejection force of <215 N, a friability of <0.16%, and a disintegration time of <580 seconds. 33. The directly compressible tabletting composition according to claim 21, wherein the composition is in the form of tablets formed by compression with a pressing force of 20 kN, and the tablets have a hardness of >300 N, together with an ejection force of <100 N, a friability of <0.16% and a disintegration time of <580 seconds. 34. The directly compressible tabletting composition according to claim 21, wherein the composition is in the form of tablets formed by compression with a pressing force of 30 kN, and the tablets have a hardness of >350 N, together with an ejection force of <115 N, a friability of <0.14% and a disintegration time of <550 seconds. 35. The directly compressible tabletting composition according to claim 21, wherein the composition is in the form of tablets formed by compression. 36. A process for preparing the directly compressible tabletting composition according to claim 21, which comprises dissolving or suspending a solution or suspension comprising 50 to 85% by weight of anhydrous calcium hydrogenphosphate, 10 to 40% by weight of mannitol and 5 to 20% by weight of sorbitol in water, where 4 parts of solid are dissolved or suspended in 4 parts of water, and subjecting the solution or suspension to a co-spray-granulation process, either batchwise or continuously, in a fluidised-bed granulator. 37. A process for preparing the directly compressible tabletting composition according to claim 26, which comprises dissolving or suspending a solution or suspension comprising 60 to 80% by weight of anhydrous calcium hydrogenphosphate, 15 to 25% by weight of mannitol and 7 to 13% by weight of sorbitol in water, where 4 parts of solid are dissolved or suspended in 4 parts of water, and subjecting the solution or suspension to a co-spray-granulation process, either batchwise or continuously, in a fluidised-bed granulator. | The present invention relates to a Tabletting aid with a low water content and to a process for the preparation thereof. The Tabletting aid composition is a directly compressible composition, the use of which results in improved tablet properties.1.-20. (canceled) 21. A directly compressible tabletting composition, comprising 50-85% by weight of anhydrous calcium hydrogenphosphate, 10-40% by weight of mannitol and 5-20% by weight of sorbitol, wherein said composition has a flow angle in the range of 29 to 33.4° and a bulk density in the range of 0.56 to 0.77 g/ml with a tapped density in the range of 0.73 to 0.92 g/ml. 22. The directly compressible tabletting composition according to claim 21, further comprising one or more homogeneously distributed, water-insoluble and/or water-soluble additives. 23. The directly compressible tabletting composition according to claim 21, one or more one or more homogeneously distributed, water-insoluble and/or water-soluble additives selected from: pharmaceutical active compounds, plant extracts, sweeteners, dyes, citric acid, vitamins and trace elements. 24. The directly compressible tabletting composition according to claim 23, which comprises one or more pharmaceutical active compounds which are analgesics. 25. The directly compressible tabletting composition according to claim 23, which comprises one or more sweeteners selected from: acesulfame K, Aspartame®, saccharin, cyclamate, sucralose and neohesperidin DC. 26. The directly compressible tabletting composition according to claim 21, which comprises 60 to 80% by weight of anhydrous calcium hydrogenphosphate, 15 to 25% by weight of mannitol and 7 to 13% by weight of sorbitol. 27. The directly compressible tabletting composition according to claim 21, which comprises 65 to 75% by weight of anhydrous calcium hydrogenphosphate, 17 to 23% by weight of mannitol and 8 to 12% by weight of sorbitol. 28. The directly compressible tabletting composition according to claim 21, wherein the anhydrous calcium hydrogenphosphate, mannitol and sorbitol are co-spray granulated. 29. The directly compressible tabletting composition according to claim 21, wherein the composition has a particle-size distribution of max. 3% by weight of undersized particles having a particle size of <32 μm, max. 5% by weight of oversized particles having a particle size of >500 μm, and 50 to 90% by weight of a particle fraction having particle sizes in the range from 100 to 315 μm. 30. The directly compressible tabletting composition according to claim 21, wherein the composition has a calcium content of 14 to 21% by weight, based on the total amount, and a drying loss of less than 2% by weight. 31. The directly compressible tabletting composition according to claim 21, wherein the composition has a calcium content of 14 to 21% by weight, based on the total amount, and a drying loss of less than 1% by weight. 32. The directly compressible tabletting composition according to claim 21, wherein the composition is in the form of tablets formed by compression with a pressing force of 20 kN, and the tablets have a hardness of >270 N, together with an ejection force of <215 N, a friability of <0.16%, and a disintegration time of <580 seconds. 33. The directly compressible tabletting composition according to claim 21, wherein the composition is in the form of tablets formed by compression with a pressing force of 20 kN, and the tablets have a hardness of >300 N, together with an ejection force of <100 N, a friability of <0.16% and a disintegration time of <580 seconds. 34. The directly compressible tabletting composition according to claim 21, wherein the composition is in the form of tablets formed by compression with a pressing force of 30 kN, and the tablets have a hardness of >350 N, together with an ejection force of <115 N, a friability of <0.14% and a disintegration time of <550 seconds. 35. The directly compressible tabletting composition according to claim 21, wherein the composition is in the form of tablets formed by compression. 36. A process for preparing the directly compressible tabletting composition according to claim 21, which comprises dissolving or suspending a solution or suspension comprising 50 to 85% by weight of anhydrous calcium hydrogenphosphate, 10 to 40% by weight of mannitol and 5 to 20% by weight of sorbitol in water, where 4 parts of solid are dissolved or suspended in 4 parts of water, and subjecting the solution or suspension to a co-spray-granulation process, either batchwise or continuously, in a fluidised-bed granulator. 37. A process for preparing the directly compressible tabletting composition according to claim 26, which comprises dissolving or suspending a solution or suspension comprising 60 to 80% by weight of anhydrous calcium hydrogenphosphate, 15 to 25% by weight of mannitol and 7 to 13% by weight of sorbitol in water, where 4 parts of solid are dissolved or suspended in 4 parts of water, and subjecting the solution or suspension to a co-spray-granulation process, either batchwise or continuously, in a fluidised-bed granulator. | 1,600 |
1,043 | 15,909,314 | 1,656 | Compositions and methods relating to ApoA-1 fusion polypeptides are disclosed. The fusion polypeptides include a first polypeptide segment corresponding to an ApoA-1 polypeptide or ApoA-1 mimetic, and may also include a dimerizing domain such as, e.g., an Fc region, which is typically linked carboxyl-terminal to the first polypeptide segment via a flexible linker. In some embodiments, the fusion polypeptide further includes a second polypeptide segment located carboxyl-terminal to the first polypeptide segment and which confers a second biological activity (e.g., an RNase, paraoxonase, platelet-activating factor acetylhydrolase, cholesterol ester transfer protein, lecithin-cholesterol acyltransferase, polypeptide that specifically binds to proprotein convertase subtilisin/kexin type 9, or polypeptide that specifically binds to amyloid beta). Also disclosed are dimeric proteins comprising first and second ApoA-1 fusion polypeptides as disclosed herein. The fusion polypeptides and dimeric proteins are useful in methods for therapy. | 1. A fusion polypeptide comprising, from an amino-terminal position to a carboxyl-terminal position, ApoA1-L1-D, wherein:
ApoA1 is a first polypeptide segment comprising an amino acid sequence having at least 95% identity with amino acid residues 19-267 or 25-267 of SEQ ID NO:2, wherein said first polypeptide segment has cholesterol efflux activity; L1 is a first polypeptide linker comprising at least 10 amino acid residues; and D is an immunoglobulin Fc region. 2. The fusion polypeptide of claim 1, wherein L1 comprises at least 16 amino acid residues. 3. The fusion polypeptide of claim 1, wherein L1 consists of from 16 to 60 amino acid residues. 4. The fusion polypeptide of claim 1, wherein L1 comprises two or more tandem repeats of the amino acid sequence of SEQ ID NO:15. 5. The fusion polypeptide of claim 1, wherein the first polypeptide segment has the amino acid sequence shown in residues 19-267 or 25-267 of SEQ ID NO:2. 6. The fusion polypeptide of claim 1, wherein the Fc region is a human Fc region 7. The fusion polypeptide of claim 6, wherein the human Fc region is an Fc variant comprising one or more amino acid substitutions relative to the wild-type human sequence. 8. The fusion polypeptide of claim 6, wherein the Fc region is a human γ1 Fc region or a human γ3 Fc region. 9. The fusion polypeptide of claim 7, wherein the Fc region is a human γ1 Fc variant in which Eu residues C220, C226, and C229 are each replaced by serine. 10. The fusion polypeptide of claim 9, wherein Eu residue P238 is replaced by serine. 11. The fusion polypeptide of claim 8, wherein the Fc region is a human γ1 Fc variant in which Eu residue P331 is replaced by serine. 12. The fusion polypeptide of claim 10, wherein Eu residue P331 is replaced by serine. 13. The fusion polypeptide of claim 1, wherein the Fc region has the amino acid sequence shown in (i) residues 294-525 or 294-524 of SEQ ID NO:2, or (ii) residues 294-525 or 294-524 of SEQ ID NO:13. 14. The fusion polypeptide of claim 1, wherein the fusion polypeptide comprises an amino acid sequence having at least 95% identity with
(i) residues 19-525, 19-524, 25-525, or 25-524 of SEQ ID NO:2, (ii) residues 19-525, 19-524, 25-525, or 25-524 of SEQ ID NO:13, (iii) residues 19-515, 19-514, 25-515, or 25-514 of SEQ ID NO:22, (iv) residues 19-520, 19-519, 25-520, or 25-519 of SEQ ID NO:26, or (v) residues 19-535, 19-534, 25-535, or 25-534 of SEQ ID NO:24. 15. The fusion polypeptide of claim 14, wherein the fusion polypeptide comprises the amino acid sequence shown in
(i) residues 19-525, 19-524, 25-525, or 25-524 of SEQ ID NO:2, (ii) residues 19-525, 19-524, 25-525, or 25-524 of SEQ ID NO:13, (iii) residues 19-515, 19-514, 25-515, or 25-514 of SEQ ID NO:22, (iv) residues 19-520, 19-519, 25-520, or 25-519 of SEQ ID NO:26, or (v) residues 19-535, 19-534, 25-535, or 25-534 of SEQ ID NO:24. 16. The fusion polypeptide of claim 1, further comprising a second polypeptide segment located carboxyl-terminal to the immunoglobulin Fc region, wherein the second polypeptide segment is selected from the group consisting of an RNase and a paraoxonase. 17. The fusion polypeptide of claim 16, wherein the second polypeptide segment is the RNase. 18. The fusion polypeptide of claim 17, wherein the RNase has at least 95% identity with amino acid residues 544-675 or 548-675 of SEQ ID NO:4. 19. The fusion polypeptide of claim 18, wherein the RNase has the amino acid sequence shown in residues 544-675 or 548-675 of SEQ ID NO:4. 20. The fusion polypeptide of claim 17, wherein the fusion polypeptide comprises an amino acid sequence having at least 95% identity with
(i) residues 19-675 or 25-675 of SEQ ID NO:4, (ii) residues 19-675 or 25-675 of SEQ ID NO:14, (iii) residues 19-671 or 25-671 of SEQ ID NO:58, or (iv) residues 19-671 or 25-671 of SEQ ID NO:59. 21. The fusion polypeptide of claim 20, wherein the fusion polypeptide comprises the amino acid sequence shown in
(i) residues 19-675 or 25-675 of SEQ ID NO:4, (ii) residues 19-675 or 25-675 of SEQ ID NO:14, (iii) residues 19-671 or 25-671 of SEQ ID NO:58, or (iv) residues 19-671 or 25-671 of SEQ ID NO:59. 22. The fusion polypeptide of claim 16, wherein the second polypeptide segment is the paraoxonase. 23. The fusion polypeptide of claim 22, wherein the paraoxonase has at least 95% identity with amino acid residues 16-355 of SEQ ID NO:12, amino acid residues 16-355 of SEQ ID NO:42, or amino acid residues 16-355 of SEQ ID NO:44. 24. The fusion polypeptide of claim 23, wherein the paraoxonase has the amino acid sequence shown in residues 16-355 of SEQ ID NO:12, residues 16-355 of SEQ ID NO:42, or residues 16-355 of SEQ ID NO:44. 25. The fusion polypeptide of claim 22, wherein the fusion polypeptide comprises an amino acid sequence having at least 95% identity with
(i) residues 19-883 or 25-883 of SEQ ID NO:28, (ii) residues 19-873 or 25-873 of SEQ ID NO:38, (iii) residues 19-883 or 25-883 of SEQ ID NO:46, or (iv) residues 19-883 or 25-883 of SEQ ID NO:48. 26. The fusion polypeptide of claim 25, wherein the fusion polypeptide comprises the amino acid sequence shown in
(i) residues 19-883 or 25-883 of SEQ ID NO:28, (ii) residues 19-873 or 25-873 of SEQ ID NO:38, (iii) residues 19-883 or 25-883 of SEQ ID NO:46, or (iv) residues 19-883 or 25-883 of SEQ ID NO:48. 27. A dimeric protein comprising a first fusion polypeptide and a second fusion polypeptide, wherein each of said first and second fusion polypeptides is a fusion polypeptide as defined in claim 1. 28. A polynucleotide encoding the fusion polypeptide of claim 1. 29. A method of making a fusion polypeptide, the method comprising:
culturing a cell into which has been introduced an expression vector comprising the following operably linked elements:
a transcription promoter,
a DNA segment encoding the fusion polypeptide of claim 1, and
a transcription terminator,
wherein the cell expresses the DNA segment and the encoded fusion polypeptide is produced; and
recovering the fusion polypeptide. 30. The method of claim 29, wherein the encoded fusion polypeptide is produced in the cell and recovered as a dimeric protein. | Compositions and methods relating to ApoA-1 fusion polypeptides are disclosed. The fusion polypeptides include a first polypeptide segment corresponding to an ApoA-1 polypeptide or ApoA-1 mimetic, and may also include a dimerizing domain such as, e.g., an Fc region, which is typically linked carboxyl-terminal to the first polypeptide segment via a flexible linker. In some embodiments, the fusion polypeptide further includes a second polypeptide segment located carboxyl-terminal to the first polypeptide segment and which confers a second biological activity (e.g., an RNase, paraoxonase, platelet-activating factor acetylhydrolase, cholesterol ester transfer protein, lecithin-cholesterol acyltransferase, polypeptide that specifically binds to proprotein convertase subtilisin/kexin type 9, or polypeptide that specifically binds to amyloid beta). Also disclosed are dimeric proteins comprising first and second ApoA-1 fusion polypeptides as disclosed herein. The fusion polypeptides and dimeric proteins are useful in methods for therapy.1. A fusion polypeptide comprising, from an amino-terminal position to a carboxyl-terminal position, ApoA1-L1-D, wherein:
ApoA1 is a first polypeptide segment comprising an amino acid sequence having at least 95% identity with amino acid residues 19-267 or 25-267 of SEQ ID NO:2, wherein said first polypeptide segment has cholesterol efflux activity; L1 is a first polypeptide linker comprising at least 10 amino acid residues; and D is an immunoglobulin Fc region. 2. The fusion polypeptide of claim 1, wherein L1 comprises at least 16 amino acid residues. 3. The fusion polypeptide of claim 1, wherein L1 consists of from 16 to 60 amino acid residues. 4. The fusion polypeptide of claim 1, wherein L1 comprises two or more tandem repeats of the amino acid sequence of SEQ ID NO:15. 5. The fusion polypeptide of claim 1, wherein the first polypeptide segment has the amino acid sequence shown in residues 19-267 or 25-267 of SEQ ID NO:2. 6. The fusion polypeptide of claim 1, wherein the Fc region is a human Fc region 7. The fusion polypeptide of claim 6, wherein the human Fc region is an Fc variant comprising one or more amino acid substitutions relative to the wild-type human sequence. 8. The fusion polypeptide of claim 6, wherein the Fc region is a human γ1 Fc region or a human γ3 Fc region. 9. The fusion polypeptide of claim 7, wherein the Fc region is a human γ1 Fc variant in which Eu residues C220, C226, and C229 are each replaced by serine. 10. The fusion polypeptide of claim 9, wherein Eu residue P238 is replaced by serine. 11. The fusion polypeptide of claim 8, wherein the Fc region is a human γ1 Fc variant in which Eu residue P331 is replaced by serine. 12. The fusion polypeptide of claim 10, wherein Eu residue P331 is replaced by serine. 13. The fusion polypeptide of claim 1, wherein the Fc region has the amino acid sequence shown in (i) residues 294-525 or 294-524 of SEQ ID NO:2, or (ii) residues 294-525 or 294-524 of SEQ ID NO:13. 14. The fusion polypeptide of claim 1, wherein the fusion polypeptide comprises an amino acid sequence having at least 95% identity with
(i) residues 19-525, 19-524, 25-525, or 25-524 of SEQ ID NO:2, (ii) residues 19-525, 19-524, 25-525, or 25-524 of SEQ ID NO:13, (iii) residues 19-515, 19-514, 25-515, or 25-514 of SEQ ID NO:22, (iv) residues 19-520, 19-519, 25-520, or 25-519 of SEQ ID NO:26, or (v) residues 19-535, 19-534, 25-535, or 25-534 of SEQ ID NO:24. 15. The fusion polypeptide of claim 14, wherein the fusion polypeptide comprises the amino acid sequence shown in
(i) residues 19-525, 19-524, 25-525, or 25-524 of SEQ ID NO:2, (ii) residues 19-525, 19-524, 25-525, or 25-524 of SEQ ID NO:13, (iii) residues 19-515, 19-514, 25-515, or 25-514 of SEQ ID NO:22, (iv) residues 19-520, 19-519, 25-520, or 25-519 of SEQ ID NO:26, or (v) residues 19-535, 19-534, 25-535, or 25-534 of SEQ ID NO:24. 16. The fusion polypeptide of claim 1, further comprising a second polypeptide segment located carboxyl-terminal to the immunoglobulin Fc region, wherein the second polypeptide segment is selected from the group consisting of an RNase and a paraoxonase. 17. The fusion polypeptide of claim 16, wherein the second polypeptide segment is the RNase. 18. The fusion polypeptide of claim 17, wherein the RNase has at least 95% identity with amino acid residues 544-675 or 548-675 of SEQ ID NO:4. 19. The fusion polypeptide of claim 18, wherein the RNase has the amino acid sequence shown in residues 544-675 or 548-675 of SEQ ID NO:4. 20. The fusion polypeptide of claim 17, wherein the fusion polypeptide comprises an amino acid sequence having at least 95% identity with
(i) residues 19-675 or 25-675 of SEQ ID NO:4, (ii) residues 19-675 or 25-675 of SEQ ID NO:14, (iii) residues 19-671 or 25-671 of SEQ ID NO:58, or (iv) residues 19-671 or 25-671 of SEQ ID NO:59. 21. The fusion polypeptide of claim 20, wherein the fusion polypeptide comprises the amino acid sequence shown in
(i) residues 19-675 or 25-675 of SEQ ID NO:4, (ii) residues 19-675 or 25-675 of SEQ ID NO:14, (iii) residues 19-671 or 25-671 of SEQ ID NO:58, or (iv) residues 19-671 or 25-671 of SEQ ID NO:59. 22. The fusion polypeptide of claim 16, wherein the second polypeptide segment is the paraoxonase. 23. The fusion polypeptide of claim 22, wherein the paraoxonase has at least 95% identity with amino acid residues 16-355 of SEQ ID NO:12, amino acid residues 16-355 of SEQ ID NO:42, or amino acid residues 16-355 of SEQ ID NO:44. 24. The fusion polypeptide of claim 23, wherein the paraoxonase has the amino acid sequence shown in residues 16-355 of SEQ ID NO:12, residues 16-355 of SEQ ID NO:42, or residues 16-355 of SEQ ID NO:44. 25. The fusion polypeptide of claim 22, wherein the fusion polypeptide comprises an amino acid sequence having at least 95% identity with
(i) residues 19-883 or 25-883 of SEQ ID NO:28, (ii) residues 19-873 or 25-873 of SEQ ID NO:38, (iii) residues 19-883 or 25-883 of SEQ ID NO:46, or (iv) residues 19-883 or 25-883 of SEQ ID NO:48. 26. The fusion polypeptide of claim 25, wherein the fusion polypeptide comprises the amino acid sequence shown in
(i) residues 19-883 or 25-883 of SEQ ID NO:28, (ii) residues 19-873 or 25-873 of SEQ ID NO:38, (iii) residues 19-883 or 25-883 of SEQ ID NO:46, or (iv) residues 19-883 or 25-883 of SEQ ID NO:48. 27. A dimeric protein comprising a first fusion polypeptide and a second fusion polypeptide, wherein each of said first and second fusion polypeptides is a fusion polypeptide as defined in claim 1. 28. A polynucleotide encoding the fusion polypeptide of claim 1. 29. A method of making a fusion polypeptide, the method comprising:
culturing a cell into which has been introduced an expression vector comprising the following operably linked elements:
a transcription promoter,
a DNA segment encoding the fusion polypeptide of claim 1, and
a transcription terminator,
wherein the cell expresses the DNA segment and the encoded fusion polypeptide is produced; and
recovering the fusion polypeptide. 30. The method of claim 29, wherein the encoded fusion polypeptide is produced in the cell and recovered as a dimeric protein. | 1,600 |
1,044 | 12,646,839 | 1,634 | Methods for detecting and assessing the progress and prognosis of gastric cancer are presented. Primers and probes for use in the methods are also disclosed as are methods for suppressing gastric cancer and kits for implementing the methods. | 1. A method for detecting gastric cancer in a biological sample, comprising the step of:
detecting methylation of a patient sample sequence of at least 15 consecutive base pairs, within a contiguous sequence at least 95% similar to the region consisting of SEQ ID NO:1; wherein significant methylation level is indicative of cancer presence in the sample. 2. The method according to claim 1 wherein the target sequence is at least 50 base pairs long and contains a plurality of CpG base pairs. 3. The method according to claim 1 further comprising comparing the methylation level of the patient sample DNA with methylation level of non-cancerous cells. 4. The method according to claim 1, wherein said determining comprises treating the sample with a reagent that differentially modifies methylated and unmethylated DNA. 5. The method according to claim 4, wherein the reagent comprises a restriction enzyme that preferentially cleaves unmethylated DNA. 6. The method according to claim 1 wherein said determining comprises treating the sample with sodium bisulphate. 7. The method according to claim 4, wherein the determination is performed by combined bisulfite restriction analysis (COBRA). 8. The method according to claim 1 wherein the sample is a blood sample. 9. The method according to claim 1 wherein said determining comprises the steps of:
a) amplifying DNA treated with a restriction enzyme using primers selective for a CpG-containing genomic sequence, wherein the genomic sequence is contained within SEQ ID NO:1; and
b) comparing the level of the amplified portion of the genomic sequence in unknown samples to compare the methylation level with non-cancerous sample to thereby detect the presence of gastric cancer. 10. The method according to claim 3, wherein the reagent comprises an enzyme that preferentially cleaves unmethylated DNA. 11. The method according to claim 10, wherein said amplifying uses the polymerase chain reaction. 12. The method according to claim 1 wherein said detecting uses a primer or probe selected from the group consisting of: SEQ ID NOS:3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 35, 36, 43, 44 and 46. 13. A method for detecting gastric cancer in a biological sample, said method comprising the step of:
a) detecting the level of a target RNA with at least 95% sequence similarity to a region of at least 15 contiguous bases of sequence SEQ ID NO:2 contained in the sample, b) wherein a significantly lower amount of the said sequence in the sample relative to a non-cancerous control sample is indicative of presence of gastric cancer in the biological sample. 14. The method according to claim 13 wherein the region is at least 25 base pairs long. 15. The method according to claim 13 wherein the sample comprises gastric tissue. 16. The method according to claim 13 wherein said detecting comprises amplifying said region. 17. The method according to claim 13 wherein said amplifying uses the polymerase chain reaction. 18. The method according to claim 13 wherein said detecting comprises using a primer or probe selected from the group consisting of: SEQ ID NOS:28, 29, 31, 32, 33, 34, 37 and 38. 19. An isolated nucleic acid sequence at least about 10 base pairs long and 95% identical to a fragment of SEQ ID NO:1, or SEQ ID NO:2. 20. The isolated nucleic acid according to claim 19 wherein said isolated sequence is at least about 20 base pairs long and at least 99% similar to the corresponding fragment of said region. | Methods for detecting and assessing the progress and prognosis of gastric cancer are presented. Primers and probes for use in the methods are also disclosed as are methods for suppressing gastric cancer and kits for implementing the methods.1. A method for detecting gastric cancer in a biological sample, comprising the step of:
detecting methylation of a patient sample sequence of at least 15 consecutive base pairs, within a contiguous sequence at least 95% similar to the region consisting of SEQ ID NO:1; wherein significant methylation level is indicative of cancer presence in the sample. 2. The method according to claim 1 wherein the target sequence is at least 50 base pairs long and contains a plurality of CpG base pairs. 3. The method according to claim 1 further comprising comparing the methylation level of the patient sample DNA with methylation level of non-cancerous cells. 4. The method according to claim 1, wherein said determining comprises treating the sample with a reagent that differentially modifies methylated and unmethylated DNA. 5. The method according to claim 4, wherein the reagent comprises a restriction enzyme that preferentially cleaves unmethylated DNA. 6. The method according to claim 1 wherein said determining comprises treating the sample with sodium bisulphate. 7. The method according to claim 4, wherein the determination is performed by combined bisulfite restriction analysis (COBRA). 8. The method according to claim 1 wherein the sample is a blood sample. 9. The method according to claim 1 wherein said determining comprises the steps of:
a) amplifying DNA treated with a restriction enzyme using primers selective for a CpG-containing genomic sequence, wherein the genomic sequence is contained within SEQ ID NO:1; and
b) comparing the level of the amplified portion of the genomic sequence in unknown samples to compare the methylation level with non-cancerous sample to thereby detect the presence of gastric cancer. 10. The method according to claim 3, wherein the reagent comprises an enzyme that preferentially cleaves unmethylated DNA. 11. The method according to claim 10, wherein said amplifying uses the polymerase chain reaction. 12. The method according to claim 1 wherein said detecting uses a primer or probe selected from the group consisting of: SEQ ID NOS:3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 35, 36, 43, 44 and 46. 13. A method for detecting gastric cancer in a biological sample, said method comprising the step of:
a) detecting the level of a target RNA with at least 95% sequence similarity to a region of at least 15 contiguous bases of sequence SEQ ID NO:2 contained in the sample, b) wherein a significantly lower amount of the said sequence in the sample relative to a non-cancerous control sample is indicative of presence of gastric cancer in the biological sample. 14. The method according to claim 13 wherein the region is at least 25 base pairs long. 15. The method according to claim 13 wherein the sample comprises gastric tissue. 16. The method according to claim 13 wherein said detecting comprises amplifying said region. 17. The method according to claim 13 wherein said amplifying uses the polymerase chain reaction. 18. The method according to claim 13 wherein said detecting comprises using a primer or probe selected from the group consisting of: SEQ ID NOS:28, 29, 31, 32, 33, 34, 37 and 38. 19. An isolated nucleic acid sequence at least about 10 base pairs long and 95% identical to a fragment of SEQ ID NO:1, or SEQ ID NO:2. 20. The isolated nucleic acid according to claim 19 wherein said isolated sequence is at least about 20 base pairs long and at least 99% similar to the corresponding fragment of said region. | 1,600 |
1,045 | 16,133,359 | 1,612 | Some embodiments described herein provide for a multi-component compositions and methods for its pharmaceutical and cosmetic use, comprising a combination of an aliphatic anionic compound, an oxidative compound, and a buffering system. Source of fluoride ion and other carriers are optional ingredients. The aliphatic anionic compound and the oxidative compound function together, in presence or absence of fluoride ion source, to protect the oxidative compounds from degradation prior to use and upon use, and to enhance the efficacy of the composition. In addition to achieving greater stability, combined effects of the aliphatic anionic compound, oxidative compound and source of fluoride ion achieve enhanced fluoride uptake, higher enamel protection by enhanced remineralization and reduced demineralization, increased plaque removal, reduced re-growth of plaque polymicrobial biofilm, greater amount of chlorite ion availability and effective oxidation of salivary biomolecules. | 1. A multi-component composition comprising:
from about 0.01% to about 5.0% of an aliphatic anionic compound, based on a total weight of the multi-component composition; from about 0.001 to about 8.0% of an oxidative compound, based on a total weight of the multi-component composition; a buffering system, wherein pH of the multi-component composition is between 6.0 and 8.0; and water, wherein the aliphatic anionic compound provides enhanced stability for the oxidative compound. 2. The multi-component composition of claim 1, wherein the aliphatic anionic compound comprises, at least one of, N-acyl sarcosinate, taurates, sodium lauryl sulfoacetate, sodium lauryl isethionate, sodium laureth carboxylate. 3. The multi-component composition of claim 2, wherein the N-acyl sarcosinate is, at least one of, sodium lauroyl sarcosinate, sodium cocoyl sarcosinate, sodium myristoyl sarcosinate, sodium oleoyl sarcosinate, or sodium stearoyl sarcosinate. 4. The multi-component composition of claim 1, wherein the oxidative compound comprises, at least one of, ammonium peroxydisulfate, carbamide (urea) peroxide, ferric chloride, hydrogen peroxide, potassium bromate, potassium chlorate, potassium perchlorate, potassium dichromate, potassium ferricyanide, potassium peroxymonosulfate, potassium persulfate, sodium bromate, sodium chlorate, sodium perchlorate, sodium chlorite, sodium hypochlorite, sodium iodate, sodium perborate, sodium percarbonate, sodium persulfate, stabilized chlorine dioxide, strontium peroxide, zinc chloride and zinc peroxide. 5. The multi-component composition of claim 1, wherein the multi-component composition is formulated into a form, at least one of, a mouth rinse, a gum, a gel, a paste, a cream, spray, and a lozenge. 6. The multi-component composition of claim 1, further comprising an orally acceptable aqueous vehicle comprising, at least one, a humectant, an abrasive, a pharmaceutically acceptable carrier, a fluoride ion source, and a thickening agent. 7. The multi-component composition of claim 1, wherein the multi-component composition oxidizes salivary biomolecules. 8. The multi-component composition of claim 7, wherein the multi-component composition oxidizes salivary biomolecules in 30 to 120 seconds of contact with saliva. 9. The multi-component composition of claim 8, wherein the salivary biomolecules are pyruvate and L-methionine. 10. The multi-component composition of claim 1, wherein the multi-component composition is applied to, at least one of, anal, aural, nasal, oral, and urogenital cavities. 11. The multi-component composition of claim 1, wherein less than 20% of the oxidative compound is destabilized after 3 months at 40±1° C. and 70-75% relative humidity or one year under ambient conditions. 12. The multi-component composition of claim 1, wherein the oxidative compound is stabilized chlorine dioxide, wherein less than 20% of the stabilized chlorine dioxide is degraded after 3 months at 40±1° C. and 70-75% relative humidity or one year under ambient conditions. 13. An oral care composition, comprising:
from about 0.01% to about 5.0% of an N-acyl sarcosinate, based on a total weight of the oral care composition; from about 0.001 to about 8% of an oxidative compound, based on the total weight of the oral care composition; a buffering system, wherein pH of the multi-component composition is between 6.0 and 8.0; and water, wherein the N-acyl sarcosinate provides enhanced stability for the oxidative compound in the oral care composition. 14. The oral care composition of claim 13, wherein the N-acyl sarcosinate is selected from sodium lauroyl sarcosinate, sodium cocoyl sarcosinate, or sodium myristoyl sarcosinate. 15. The oral care composition of claim 13, wherein the oxidative compound is selected from sodium chlorite or stabilized chlorine dioxide. 16. The oral care composition of claim 13, further comprising at least one material selected from a group consisting of a humectant, a whitening agent, a thickening agent, a fluoride ion source, a sweetening agent, an abrasive, a flavoring agent, a coloring agent, and a gelling agent. 17. The oral care composition of claim 13, wherein the oral care composition is a dentifrice. 18. The oral care composition of claim 13, wherein less than 20% of the oxidative compound is degraded in 3 months at 40±1° C. and 70-75% relative humidity or one year under ambient conditions. 19. The oral care composition of claim 13, wherein the buffering system comprises disodium hydrogen phosphate and sodium dihydrogen phosphate. 20. The oral care composition of claim 13, wherein the oral care composition decreases regrowth of an oral polymicrobial biofilm. 21. The oral care composition of claim 13, wherein the oral care composition effectively reduces regrowth of the oral polymicrobial biofilm in 24 hours. 22. The oral care composition of claim 13, wherein the oral care composition enhances remineralization of a tooth enamel. 23. The oral care composition of claim 13, wherein the oral care composition oxidizes salivary biomolecules such as pyruvate and L-methionine in 30 to 60 seconds of contact with the saliva in the oral cavity. 24. The oral care composition of claim 13, wherein the oral care composition provides a greater amount of available chlorite ion. 25. A method for effecting enhanced fluoride uptake into an oral cavity, comprising:
preparing an oral care composition comprising from about 0.01% to about 5.0% of an N-acyl sarcosinate, based on a total weight of the oral care composition, from about 0.001 to about 8.0% of an oxidative compound, based on the total weight of the oral care composition, a buffering system, wherein pH of the multi-component composition is between 6.0 and 8.0, water; and applying the oral care composition to the oral cavity. 26. The method of claim 25, wherein the oral care composition enhances fluoride uptake in the oral cavity. 27. The method of claim 25, wherein the enhanced fluoride uptake into the oral cavity is increased by at least 2-fold. 28. The method of claim 25, wherein the enhanced fluoride uptake into the oral cavity is increased by at least 4-fold. 29. A method to decrease regrowth of oral polymicrobial biofilm, comprising:
preparing an oral care composition comprising from about 0.01% to about 5.0% of an N-acyl sarcosinate, based on a total weight of the oral care composition, from about 0.001 to about 8.0% of an oxidative compound, based on the total weight of the oral care composition, a buffering system, wherein pH of the multi-component composition is between 6.0 and 8.0, water; and applying the oral care composition to an oral cavity. 30. The method of claim 29, wherein the oral care composition decreases oral polymicrobial biofilm. | Some embodiments described herein provide for a multi-component compositions and methods for its pharmaceutical and cosmetic use, comprising a combination of an aliphatic anionic compound, an oxidative compound, and a buffering system. Source of fluoride ion and other carriers are optional ingredients. The aliphatic anionic compound and the oxidative compound function together, in presence or absence of fluoride ion source, to protect the oxidative compounds from degradation prior to use and upon use, and to enhance the efficacy of the composition. In addition to achieving greater stability, combined effects of the aliphatic anionic compound, oxidative compound and source of fluoride ion achieve enhanced fluoride uptake, higher enamel protection by enhanced remineralization and reduced demineralization, increased plaque removal, reduced re-growth of plaque polymicrobial biofilm, greater amount of chlorite ion availability and effective oxidation of salivary biomolecules.1. A multi-component composition comprising:
from about 0.01% to about 5.0% of an aliphatic anionic compound, based on a total weight of the multi-component composition; from about 0.001 to about 8.0% of an oxidative compound, based on a total weight of the multi-component composition; a buffering system, wherein pH of the multi-component composition is between 6.0 and 8.0; and water, wherein the aliphatic anionic compound provides enhanced stability for the oxidative compound. 2. The multi-component composition of claim 1, wherein the aliphatic anionic compound comprises, at least one of, N-acyl sarcosinate, taurates, sodium lauryl sulfoacetate, sodium lauryl isethionate, sodium laureth carboxylate. 3. The multi-component composition of claim 2, wherein the N-acyl sarcosinate is, at least one of, sodium lauroyl sarcosinate, sodium cocoyl sarcosinate, sodium myristoyl sarcosinate, sodium oleoyl sarcosinate, or sodium stearoyl sarcosinate. 4. The multi-component composition of claim 1, wherein the oxidative compound comprises, at least one of, ammonium peroxydisulfate, carbamide (urea) peroxide, ferric chloride, hydrogen peroxide, potassium bromate, potassium chlorate, potassium perchlorate, potassium dichromate, potassium ferricyanide, potassium peroxymonosulfate, potassium persulfate, sodium bromate, sodium chlorate, sodium perchlorate, sodium chlorite, sodium hypochlorite, sodium iodate, sodium perborate, sodium percarbonate, sodium persulfate, stabilized chlorine dioxide, strontium peroxide, zinc chloride and zinc peroxide. 5. The multi-component composition of claim 1, wherein the multi-component composition is formulated into a form, at least one of, a mouth rinse, a gum, a gel, a paste, a cream, spray, and a lozenge. 6. The multi-component composition of claim 1, further comprising an orally acceptable aqueous vehicle comprising, at least one, a humectant, an abrasive, a pharmaceutically acceptable carrier, a fluoride ion source, and a thickening agent. 7. The multi-component composition of claim 1, wherein the multi-component composition oxidizes salivary biomolecules. 8. The multi-component composition of claim 7, wherein the multi-component composition oxidizes salivary biomolecules in 30 to 120 seconds of contact with saliva. 9. The multi-component composition of claim 8, wherein the salivary biomolecules are pyruvate and L-methionine. 10. The multi-component composition of claim 1, wherein the multi-component composition is applied to, at least one of, anal, aural, nasal, oral, and urogenital cavities. 11. The multi-component composition of claim 1, wherein less than 20% of the oxidative compound is destabilized after 3 months at 40±1° C. and 70-75% relative humidity or one year under ambient conditions. 12. The multi-component composition of claim 1, wherein the oxidative compound is stabilized chlorine dioxide, wherein less than 20% of the stabilized chlorine dioxide is degraded after 3 months at 40±1° C. and 70-75% relative humidity or one year under ambient conditions. 13. An oral care composition, comprising:
from about 0.01% to about 5.0% of an N-acyl sarcosinate, based on a total weight of the oral care composition; from about 0.001 to about 8% of an oxidative compound, based on the total weight of the oral care composition; a buffering system, wherein pH of the multi-component composition is between 6.0 and 8.0; and water, wherein the N-acyl sarcosinate provides enhanced stability for the oxidative compound in the oral care composition. 14. The oral care composition of claim 13, wherein the N-acyl sarcosinate is selected from sodium lauroyl sarcosinate, sodium cocoyl sarcosinate, or sodium myristoyl sarcosinate. 15. The oral care composition of claim 13, wherein the oxidative compound is selected from sodium chlorite or stabilized chlorine dioxide. 16. The oral care composition of claim 13, further comprising at least one material selected from a group consisting of a humectant, a whitening agent, a thickening agent, a fluoride ion source, a sweetening agent, an abrasive, a flavoring agent, a coloring agent, and a gelling agent. 17. The oral care composition of claim 13, wherein the oral care composition is a dentifrice. 18. The oral care composition of claim 13, wherein less than 20% of the oxidative compound is degraded in 3 months at 40±1° C. and 70-75% relative humidity or one year under ambient conditions. 19. The oral care composition of claim 13, wherein the buffering system comprises disodium hydrogen phosphate and sodium dihydrogen phosphate. 20. The oral care composition of claim 13, wherein the oral care composition decreases regrowth of an oral polymicrobial biofilm. 21. The oral care composition of claim 13, wherein the oral care composition effectively reduces regrowth of the oral polymicrobial biofilm in 24 hours. 22. The oral care composition of claim 13, wherein the oral care composition enhances remineralization of a tooth enamel. 23. The oral care composition of claim 13, wherein the oral care composition oxidizes salivary biomolecules such as pyruvate and L-methionine in 30 to 60 seconds of contact with the saliva in the oral cavity. 24. The oral care composition of claim 13, wherein the oral care composition provides a greater amount of available chlorite ion. 25. A method for effecting enhanced fluoride uptake into an oral cavity, comprising:
preparing an oral care composition comprising from about 0.01% to about 5.0% of an N-acyl sarcosinate, based on a total weight of the oral care composition, from about 0.001 to about 8.0% of an oxidative compound, based on the total weight of the oral care composition, a buffering system, wherein pH of the multi-component composition is between 6.0 and 8.0, water; and applying the oral care composition to the oral cavity. 26. The method of claim 25, wherein the oral care composition enhances fluoride uptake in the oral cavity. 27. The method of claim 25, wherein the enhanced fluoride uptake into the oral cavity is increased by at least 2-fold. 28. The method of claim 25, wherein the enhanced fluoride uptake into the oral cavity is increased by at least 4-fold. 29. A method to decrease regrowth of oral polymicrobial biofilm, comprising:
preparing an oral care composition comprising from about 0.01% to about 5.0% of an N-acyl sarcosinate, based on a total weight of the oral care composition, from about 0.001 to about 8.0% of an oxidative compound, based on the total weight of the oral care composition, a buffering system, wherein pH of the multi-component composition is between 6.0 and 8.0, water; and applying the oral care composition to an oral cavity. 30. The method of claim 29, wherein the oral care composition decreases oral polymicrobial biofilm. | 1,600 |
1,046 | 15,622,247 | 1,631 | Embodiments of the present invention are directed to a computer-implemented method for positive OTU identification. A non-limiting example of the computer-implemented method includes receiving, by a processor, a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs. The method also includes determining, by the processor, a true positive score for each of the plurality of OTUs based upon a Čech Complex and generating a plurality of preliminary OTUs. The method also includes determining a threshold score for the preliminary OTUs. The method also includes removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold. The method also includes retaining one of the preliminary OTUs based at least in part upon a determination that the true positive score is greater than or equal to the threshold. | 1-7. (canceled) 8. A computer program product for positive operational taxonomic unit (OTU) identification, the computer program product comprising:
a computer readable storage medium having program instructions embodied therewith, wherein the instructions are executable by a processor to cause the processor to perform a method comprising:
receiving a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs;
determining a true positive score for each of the plurality of OTUs based upon a Čech Complex and generating a plurality of preliminary OTUs;
determining a threshold score for the preliminary OTUs;
removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold; and
retaining one of the preliminary OTUs as a true positive OTU based at least in part upon a determination that the true positive score is greater than or equal to the threshold. 9. The computer program product of claim 8, wherein the method further comprises sorting the true positive OTUs. 10. The computer program product of claim 9, wherein sorting comprising sorting the preliminary OTUs based at least in part upon OTU frequency. 11. The computer program product of claim 9, wherein sorting comprising sorting the preliminary OTUs based at least in part upon OTU filtration time. 12. The computer program product of claim 8, wherein the preliminary OTUs comprise a preliminary identification of an OTU in a food sample. 13. The computer program product of claim 8, wherein the true positive score of an OTU X is
tp(X)=Σh(ΣbϵH h h×len(b))
wherein b is the h-simplex=X0X1 . . . Xh and len(b) is a bar length of b. 14. The computer program product of claim 8, wherein the threshold is determined based at least in part upon the preliminary OTUs. 15. A processing system for positive operational taxonomic unit (OTU) identification, comprising:
a processor in communication with one or more types of memory, the processor configured to:
receive a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs;
determine a true positive score for each of the plurality of OTUs based upon a Čech Complex and generate a plurality of preliminary OTUs;
determine a threshold score for the preliminary OTUs;
removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold; and
retaining one of the preliminary OTUs as a true positive OTU based at least in part upon a determination that the true positive score is greater than or equal to the threshold. 16. The processing system of claim 15, wherein the method further comprises sorting the true positive OTUs. 17. The processing system of claim 16, wherein sorting comprising sorting the true positive OTUs based at least in part upon OTU frequency. 18. The processing system of claim 16, wherein sorting comprising sorting the true positive OTUs based at least in part upon OTU filtration time. 19. The processing system of claim 15, wherein the preliminary OTU comprises a preliminary identification of an OTU in a food sample. 20. The processing system of claim 15, wherein the true positive score of an OTU X is
tp(X)=Σh(ΣbϵH h xϵb h×len(b))
wherein b is the h-simplex=X0X1 . . . Xh and len(b) is a bar length of b. | Embodiments of the present invention are directed to a computer-implemented method for positive OTU identification. A non-limiting example of the computer-implemented method includes receiving, by a processor, a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs. The method also includes determining, by the processor, a true positive score for each of the plurality of OTUs based upon a Čech Complex and generating a plurality of preliminary OTUs. The method also includes determining a threshold score for the preliminary OTUs. The method also includes removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold. The method also includes retaining one of the preliminary OTUs based at least in part upon a determination that the true positive score is greater than or equal to the threshold.1-7. (canceled) 8. A computer program product for positive operational taxonomic unit (OTU) identification, the computer program product comprising:
a computer readable storage medium having program instructions embodied therewith, wherein the instructions are executable by a processor to cause the processor to perform a method comprising:
receiving a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs;
determining a true positive score for each of the plurality of OTUs based upon a Čech Complex and generating a plurality of preliminary OTUs;
determining a threshold score for the preliminary OTUs;
removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold; and
retaining one of the preliminary OTUs as a true positive OTU based at least in part upon a determination that the true positive score is greater than or equal to the threshold. 9. The computer program product of claim 8, wherein the method further comprises sorting the true positive OTUs. 10. The computer program product of claim 9, wherein sorting comprising sorting the preliminary OTUs based at least in part upon OTU frequency. 11. The computer program product of claim 9, wherein sorting comprising sorting the preliminary OTUs based at least in part upon OTU filtration time. 12. The computer program product of claim 8, wherein the preliminary OTUs comprise a preliminary identification of an OTU in a food sample. 13. The computer program product of claim 8, wherein the true positive score of an OTU X is
tp(X)=Σh(ΣbϵH h h×len(b))
wherein b is the h-simplex=X0X1 . . . Xh and len(b) is a bar length of b. 14. The computer program product of claim 8, wherein the threshold is determined based at least in part upon the preliminary OTUs. 15. A processing system for positive operational taxonomic unit (OTU) identification, comprising:
a processor in communication with one or more types of memory, the processor configured to:
receive a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs;
determine a true positive score for each of the plurality of OTUs based upon a Čech Complex and generate a plurality of preliminary OTUs;
determine a threshold score for the preliminary OTUs;
removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold; and
retaining one of the preliminary OTUs as a true positive OTU based at least in part upon a determination that the true positive score is greater than or equal to the threshold. 16. The processing system of claim 15, wherein the method further comprises sorting the true positive OTUs. 17. The processing system of claim 16, wherein sorting comprising sorting the true positive OTUs based at least in part upon OTU frequency. 18. The processing system of claim 16, wherein sorting comprising sorting the true positive OTUs based at least in part upon OTU filtration time. 19. The processing system of claim 15, wherein the preliminary OTU comprises a preliminary identification of an OTU in a food sample. 20. The processing system of claim 15, wherein the true positive score of an OTU X is
tp(X)=Σh(ΣbϵH h xϵb h×len(b))
wherein b is the h-simplex=X0X1 . . . Xh and len(b) is a bar length of b. | 1,600 |
1,047 | 13,800,066 | 1,632 | A novel cell type has been generated that has both Th1 characteristics and cytolytic activity. These Th1/killer cells are CD4+ cells purified from peripheral blood and manipulated to have Th1 characteristics such as production of IFN-gamma combined with cytolytic activity similar to cytotoxic T-cells (CTL). The CTL activity is targeted toward diseased cells, not normal cells. The cytolytic activity of the Th1/killer cells is mediated by Granzyme B-Perforin mechanism and results in apoptotic death of diseased cells. Methods of producing and using these Th1/killer cells include isolating CD4+ cells from peripheral blood, activating the CD4+ T-cells to form Th1/killer cells and administering these Th1/killer cells with the cytolytic activity to a patient wherein the Th1/killer cells are allogeneic to the patient. | 1. A composition comprising Th1/killer cells wherein the Th1/killer cells have Th1 characteristics and cytolytic activity. 2. The composition of claim 1 wherein the cytolytic activity comprises NK characteristics. 3. The composition of claim 1 wherein the Th1/killer cells express granzyme B and perforin. 4. The composition of claim 1 wherein the Th1/killer cells express IFN-gamma. 5. The composition of claim 1 wherein the Th1/killer cells have substantially reduced or no expression of IL-4. 6. The composition of claim 1 wherein the Th1/killer cells are CD4+ cells. 7. The composition of claim 1 wherein the Th1/killer cells are formulated with cross-linking agent for CD3 and CD28 8. The composition of claim 1 wherein the Th1/killer cells are derived from normal donor peripheral blood. 9. The composition of claim 1 wherein the composition comprises clinically-relevant number of the Th1/killer cells. 10. The composition of claim 1 wherein the composition comprises at least about 1×107 cells. 11. The composition of claim 1 wherein the composition comprises at least about 1×108 cells. 12. The composition of claim 1 wherein the cytolytic activity of the Th1/killer cells specifically inactivates diseased cells and not normal cells. 13. The composition of claim 12 wherein the diseased cells comprise cancerous cells, infected cells or combinations thereof. | A novel cell type has been generated that has both Th1 characteristics and cytolytic activity. These Th1/killer cells are CD4+ cells purified from peripheral blood and manipulated to have Th1 characteristics such as production of IFN-gamma combined with cytolytic activity similar to cytotoxic T-cells (CTL). The CTL activity is targeted toward diseased cells, not normal cells. The cytolytic activity of the Th1/killer cells is mediated by Granzyme B-Perforin mechanism and results in apoptotic death of diseased cells. Methods of producing and using these Th1/killer cells include isolating CD4+ cells from peripheral blood, activating the CD4+ T-cells to form Th1/killer cells and administering these Th1/killer cells with the cytolytic activity to a patient wherein the Th1/killer cells are allogeneic to the patient.1. A composition comprising Th1/killer cells wherein the Th1/killer cells have Th1 characteristics and cytolytic activity. 2. The composition of claim 1 wherein the cytolytic activity comprises NK characteristics. 3. The composition of claim 1 wherein the Th1/killer cells express granzyme B and perforin. 4. The composition of claim 1 wherein the Th1/killer cells express IFN-gamma. 5. The composition of claim 1 wherein the Th1/killer cells have substantially reduced or no expression of IL-4. 6. The composition of claim 1 wherein the Th1/killer cells are CD4+ cells. 7. The composition of claim 1 wherein the Th1/killer cells are formulated with cross-linking agent for CD3 and CD28 8. The composition of claim 1 wherein the Th1/killer cells are derived from normal donor peripheral blood. 9. The composition of claim 1 wherein the composition comprises clinically-relevant number of the Th1/killer cells. 10. The composition of claim 1 wherein the composition comprises at least about 1×107 cells. 11. The composition of claim 1 wherein the composition comprises at least about 1×108 cells. 12. The composition of claim 1 wherein the cytolytic activity of the Th1/killer cells specifically inactivates diseased cells and not normal cells. 13. The composition of claim 12 wherein the diseased cells comprise cancerous cells, infected cells or combinations thereof. | 1,600 |
1,048 | 14,406,957 | 1,627 | The present invention generally relates to the field of dietary therapies for treating disorders associated with mitochondrial dysfunction, including epilepsy. | 1. A composition suitable for human consumption comprising a decanoic acid and octanoic acid in a ratio of at least 2:1 wt/wt. 2. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 3:1 wt/wt. 3. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 4:1 wt/wt. 4. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 5:1 wt/wt. 5. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 6:1 wt/wt. 6. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 9:1 wt/wt. 7. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 10:1 wt/wt. 8. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 20:1 wt/wt. 9. The composition according to claim 1 wherein the composition is substantially free from octanoic acid. 10. The composition according to claim 1 wherein the decanoic acid is greater than 50% of the total weight of fatty acid content. 11. The composition according to claim 1 wherein the decanoic acid is at least 60% of the total weight of fatty acid content. 12. The composition according to claim 1 wherein the decanoic acid is at least 70% of the total weight of fatty acid content. 13. The composition according to claim 1 wherein the decanoic acid is at least 80% of the total weight of fatty acid content. 14. The composition according to claim 1 wherein the decanoic acid is at least 90% of the total weight of fatty acid content. 15. The composition according to claim 1 wherein the decanoic acid is at least 99% of the total weight of fatty acid content. 16. The composition according to claim 1 wherein the composition is substantially free from mono- or poly-unsaturated fatty acids. 17. A composition suitable for human consumption which has a ketogenic ratio of 0.2:1 to 5:1 and the majority of fat is decanoic acid. 18. The composition of claim 17 comprising a ketogenic ratio of 1:1 to 4:1. 19. A composition suitable for human consumption comprising proteins, fat and carbohydrates, which provides 2500 to 3100 kJ per 100 g dry mass, and at least 50% of the fat is decanoic acid. 20. The composition of claim 17 wherein at least 60% of the fat is decanoic acid. 21. The composition according to claim 17 not including any mono- and/or polyunsaturated fatty acid. 22. The composition according to claim 1 comprising 2520 to 3780 kJ per 100 g dry mass. 23. The composition according to claim 1 wherein the weight amounts of lipid to the sum of proteins and carbohydrates is 1.5-5.0 to 1. 24. The composition according to claim 1 wherein the weight amounts of lipid to the sum of proteins and carbohydrates is 2.0-3.8 to 1. 25. The composition according to claim 1 in the form of a human food stuff. 26. The composition according to claim 1 which is in a form for delivering a dosage of at least about 5 g/l to 500 g/l decanoic acid per day. 27. The composition according to claim 1 in the form of a complete nutritional product. 28. The composition according to claim 1 in a powdered form. 29. The composition according to claim 1 in a spray dried form. 30. The composition according to claim 1 for fortifying food or drink. 31. The composition according to claim 1 in the form of a food stuff. 32. The composition according to claim 1 in the form of an oil-in-water emulsion. 33. The composition according to claim 1 wherein the composition is in a form selected from the group consisting of mayonnaise, margarine, low fat spread, a dairy product including yoghurts, a cheese spread, processed cheese, a dairy dessert, a flavoured milk, cream, a fermented milk product, cheese, butter, a condensed milk product, an ice cream mix, a soya product, pasteurised liquid egg, a bakery product, a confectionary product, confectionary bar, chocolate bar, high fat bar, liquid emulsion, spray dried powder, freeze dried powder, UHT pudding, Pasteurised pudding, gel, jelly, yoghurt and a food with a fat-based or water-containing filling. 34. A method for the treatment of a disease associated with mitochondria dysfunction comprising the step of administering a composition comprising decanoic acid to an individual having same. 35. (canceled) 36. The method according to claim 34 wherein the disease is selected from the group consisting of epilepsy, diabetes, Parkinson's disease, dementias including Alzheimer's disease, and an inborn error of mitochondrial function. 37. The method according to claim 36 wherein the treatment of epilepsy includes the control of seizures. 38. A method for the prevention of a disease associated with mitochondria dysfunction comprising administering an effective amount of decanoic acid to a patient in need of the same. 39. A method of affecting at least one characteristic selected from the group consisting of the cellular content of mitochondria, the cellular function of mitochondria, the cellular availability of mitochondria, and modulating the cellular energy supply comprising administering decanoic acid to the cell. 40-42. (canceled) 43. A method according to claim 38 wherein the disease is selected from the group consisting of epilepsy, diabetes, Parkinson's disease, dementias including Alzheimer's disease, and an inborn error of mitochondrial function. 44. The method of claim 43 wherein the treatment of epilepsy includes the control of seizures. 45. (canceled) 46. The method or use according to claim 34 in which a dosage of at least about 5 g/l to 500 g/l decanoic acid is administered per day. 47. The composition according to claim 1 in the form of a bar, pudding or drink. | The present invention generally relates to the field of dietary therapies for treating disorders associated with mitochondrial dysfunction, including epilepsy.1. A composition suitable for human consumption comprising a decanoic acid and octanoic acid in a ratio of at least 2:1 wt/wt. 2. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 3:1 wt/wt. 3. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 4:1 wt/wt. 4. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 5:1 wt/wt. 5. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 6:1 wt/wt. 6. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 9:1 wt/wt. 7. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 10:1 wt/wt. 8. The composition according to claim 1 wherein the decanoic acid to octanoic acid ratio is at least 20:1 wt/wt. 9. The composition according to claim 1 wherein the composition is substantially free from octanoic acid. 10. The composition according to claim 1 wherein the decanoic acid is greater than 50% of the total weight of fatty acid content. 11. The composition according to claim 1 wherein the decanoic acid is at least 60% of the total weight of fatty acid content. 12. The composition according to claim 1 wherein the decanoic acid is at least 70% of the total weight of fatty acid content. 13. The composition according to claim 1 wherein the decanoic acid is at least 80% of the total weight of fatty acid content. 14. The composition according to claim 1 wherein the decanoic acid is at least 90% of the total weight of fatty acid content. 15. The composition according to claim 1 wherein the decanoic acid is at least 99% of the total weight of fatty acid content. 16. The composition according to claim 1 wherein the composition is substantially free from mono- or poly-unsaturated fatty acids. 17. A composition suitable for human consumption which has a ketogenic ratio of 0.2:1 to 5:1 and the majority of fat is decanoic acid. 18. The composition of claim 17 comprising a ketogenic ratio of 1:1 to 4:1. 19. A composition suitable for human consumption comprising proteins, fat and carbohydrates, which provides 2500 to 3100 kJ per 100 g dry mass, and at least 50% of the fat is decanoic acid. 20. The composition of claim 17 wherein at least 60% of the fat is decanoic acid. 21. The composition according to claim 17 not including any mono- and/or polyunsaturated fatty acid. 22. The composition according to claim 1 comprising 2520 to 3780 kJ per 100 g dry mass. 23. The composition according to claim 1 wherein the weight amounts of lipid to the sum of proteins and carbohydrates is 1.5-5.0 to 1. 24. The composition according to claim 1 wherein the weight amounts of lipid to the sum of proteins and carbohydrates is 2.0-3.8 to 1. 25. The composition according to claim 1 in the form of a human food stuff. 26. The composition according to claim 1 which is in a form for delivering a dosage of at least about 5 g/l to 500 g/l decanoic acid per day. 27. The composition according to claim 1 in the form of a complete nutritional product. 28. The composition according to claim 1 in a powdered form. 29. The composition according to claim 1 in a spray dried form. 30. The composition according to claim 1 for fortifying food or drink. 31. The composition according to claim 1 in the form of a food stuff. 32. The composition according to claim 1 in the form of an oil-in-water emulsion. 33. The composition according to claim 1 wherein the composition is in a form selected from the group consisting of mayonnaise, margarine, low fat spread, a dairy product including yoghurts, a cheese spread, processed cheese, a dairy dessert, a flavoured milk, cream, a fermented milk product, cheese, butter, a condensed milk product, an ice cream mix, a soya product, pasteurised liquid egg, a bakery product, a confectionary product, confectionary bar, chocolate bar, high fat bar, liquid emulsion, spray dried powder, freeze dried powder, UHT pudding, Pasteurised pudding, gel, jelly, yoghurt and a food with a fat-based or water-containing filling. 34. A method for the treatment of a disease associated with mitochondria dysfunction comprising the step of administering a composition comprising decanoic acid to an individual having same. 35. (canceled) 36. The method according to claim 34 wherein the disease is selected from the group consisting of epilepsy, diabetes, Parkinson's disease, dementias including Alzheimer's disease, and an inborn error of mitochondrial function. 37. The method according to claim 36 wherein the treatment of epilepsy includes the control of seizures. 38. A method for the prevention of a disease associated with mitochondria dysfunction comprising administering an effective amount of decanoic acid to a patient in need of the same. 39. A method of affecting at least one characteristic selected from the group consisting of the cellular content of mitochondria, the cellular function of mitochondria, the cellular availability of mitochondria, and modulating the cellular energy supply comprising administering decanoic acid to the cell. 40-42. (canceled) 43. A method according to claim 38 wherein the disease is selected from the group consisting of epilepsy, diabetes, Parkinson's disease, dementias including Alzheimer's disease, and an inborn error of mitochondrial function. 44. The method of claim 43 wherein the treatment of epilepsy includes the control of seizures. 45. (canceled) 46. The method or use according to claim 34 in which a dosage of at least about 5 g/l to 500 g/l decanoic acid is administered per day. 47. The composition according to claim 1 in the form of a bar, pudding or drink. | 1,600 |
1,049 | 15,801,725 | 1,631 | Embodiments of the present invention are directed to a computer-implemented method for positive OTU identification. A non-limiting example of the computer-implemented method includes receiving, by a processor, a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs. The method also includes determining, by the processor, a true positive score for each of the plurality of OTUs based upon a C̆ech Complex and generating a plurality of preliminary OTUs. The method also includes determining a threshold score for the preliminary OTUs. The method also includes removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold. The method also includes retaining one of the preliminary OTUs based at least in part upon a determination that the true positive score is greater than or equal to the threshold. | 1. A computer-implemented method for positive operational taxonomic unit (OTU) identification, the method comprising:
receiving, by a processor, a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs; determining, by the processor, a true positive score for each of the plurality of OTUs based upon a C̆ech Complex and generating a plurality of preliminary OTUs; determining, by the processor, a threshold score for the preliminary OTUs; removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold; and retaining one of the preliminary OTUs as a true positive OTU based at least in part upon a determination that the true positive score is greater than or equal to the threshold. 2. The computer implemented method of claim 1 further comprising sorting the true positive OTUs. 3. The computer implemented method of claim 2, wherein sorting comprising sorting the true positive OTUs based at least in part upon OTU frequency. 4. The computer implemented method of claim 2, wherein sorting comprising sorting the true positive OTUs based at least in part upon OTU filtration time. 5. The computer-implemented method of claim 1, wherein the preliminary OTUs comprises a preliminary identification of an OTU in a food sample. 6. The computer-implemented method of claim 1, wherein the true positive score of an OTU X is
tp(X)=Σh(ΣbϵH h, xϵb h×len(b))
wherein b is the h-simplex=X0X1. . . Xh, and len(b) is a bar length of b. 7. The computer-implemented method of claim 1, wherein the threshold is determined based at least in part upon the preliminary OTUs. | Embodiments of the present invention are directed to a computer-implemented method for positive OTU identification. A non-limiting example of the computer-implemented method includes receiving, by a processor, a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs. The method also includes determining, by the processor, a true positive score for each of the plurality of OTUs based upon a C̆ech Complex and generating a plurality of preliminary OTUs. The method also includes determining a threshold score for the preliminary OTUs. The method also includes removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold. The method also includes retaining one of the preliminary OTUs based at least in part upon a determination that the true positive score is greater than or equal to the threshold.1. A computer-implemented method for positive operational taxonomic unit (OTU) identification, the method comprising:
receiving, by a processor, a plurality of sequencing reads for a metagenome sample and, for each of the plurality of sequencing reads, a corresponding OTU set comprising a plurality of OTUs; determining, by the processor, a true positive score for each of the plurality of OTUs based upon a C̆ech Complex and generating a plurality of preliminary OTUs; determining, by the processor, a threshold score for the preliminary OTUs; removing one of the preliminary OTUs based at least in part upon a determination that the true positive score is less than a threshold; and retaining one of the preliminary OTUs as a true positive OTU based at least in part upon a determination that the true positive score is greater than or equal to the threshold. 2. The computer implemented method of claim 1 further comprising sorting the true positive OTUs. 3. The computer implemented method of claim 2, wherein sorting comprising sorting the true positive OTUs based at least in part upon OTU frequency. 4. The computer implemented method of claim 2, wherein sorting comprising sorting the true positive OTUs based at least in part upon OTU filtration time. 5. The computer-implemented method of claim 1, wherein the preliminary OTUs comprises a preliminary identification of an OTU in a food sample. 6. The computer-implemented method of claim 1, wherein the true positive score of an OTU X is
tp(X)=Σh(ΣbϵH h, xϵb h×len(b))
wherein b is the h-simplex=X0X1. . . Xh, and len(b) is a bar length of b. 7. The computer-implemented method of claim 1, wherein the threshold is determined based at least in part upon the preliminary OTUs. | 1,600 |
1,050 | 13,981,789 | 1,619 | A simple cellulose sulphate based microencapsulation technology has been applied to encapsulate bacterial or other microbial cells, which produce and release digestive enzymes and thereby provides an acid resistant shelter for these microbial cells. Surprisingly, the resulting spheres were found to provide sufficient protection for encapsulated cells from treatment with aqueous acidic solutions. Thereby the cellulose sulphate microencapsulated cells, such as probiotics are now enabled to survive passage, for example, through the stomach after consumption by a human or animal with a higher survival rate than those not within a microcapsule. After passing the stomach these cells are delivering products produced by them, e.g. enzymes or other nutrition factors. This technology therefore proves to be very useful in providing digestive or otherwise beneficial enzymes and/or of living microbial cells, into the lower gastrointestinal tract, where they could confer their health benefit to the host. | 1. Encapsulated microbial cells, wherein the microbial cells are encapsulated in a microcapsule having a porous capsule wall, wherein the porous capsule wall comprises a complex formed from sodium cellulose sulphate and poly[dimethyldiallyl-ammonium chloride], and wherein the microcapsule protects the microbial cells from being degraded by acidic aqueous solution. 2. The encapsulated microbial cells according to claim 1 wherein the cells produce and excrete digestive enzymes, and wherein the porous capsule wall is permeable to said digestive enzymes. 3. The encapsulated microbial cells according to claim 1, wherein the acidic aqueous solution is simulated gastric juice, gastric juice or gastric acid. 4. The encapsulated microbial cells according to claim 2, wherein the microcapsules release the enzymes generated by the microbial cells upon treatment with intestinal fluid or duodenal fluid. 5. The encapsulated microbial cells according to claim 4, wherein the intestinal fluid is simulated intestinal fluid (SIF) and the duodenal fluid is simulated duodenal fluid (SDF). 6. The encapsulated microbial cells according to claim 1, wherein the majority of encapsulated microbial cells survive a treatment with acidic aqueous solution according to claim 3 having a pH range between 1.0 and 3.0, preferably between 1.5 and 2.5, most preferably of 2.0 for at least 1.5 hours, preferably for at least 2.5, and more preferably for at least 4 hours. 7. The encapsulated microbial cells according to claim 6, wherein the majority of microbial cells survive a passage through the stomach of an animal. 8. The encapsulated microbial cells according to claim 7, wherein the majority is defined as a value selected from the group consisting of at least 51% of the cells, as 60% to 90% of the cells, as 60% to 80% of the cells and as 60% of the cells. 9. (canceled) 10. (canceled) 11. (canceled) 12. The encapsulated microbial cells according to claim 1, wherein the microbial cells are at least partially released in the gut of an animal. 13. The encapsulated microbial cells according to claim 1, wherein the animal is a mammal or an avian. 14. (canceled) 15. (canceled) 16. The encapsulated microbial cells according to claim 1 wherein the microcapsules have a diameter of between 0.01 and 5 mm, preferably between 0.05 and 3 mm, and most preferably between 0.1 and 1 mm. 17. The encapsulated microbial cells according to claim 1, wherein the surface pores of the porous capsule wall have a molecular weight cut off between 50 and 200 kDa, preferably between 60-150 kDa and most preferably between 60 and 100 kDa. 18. The encapsulated microbial cells according to claim 1, wherein the microbial cells are selected from the group comprising bacterial cells, yeast cells, fungal cells and probiotic cells. 19. (canceled) 20. (canceled) 21. (canceled) 22. The encapsulated bacterial cells according to claim 18, wherein the bacterial cells are selected from the group comprising Bifidobacterium, Bacteroides, Clostridium, Fusobacterium, Melissococcus, Propionibacterium, Streptococcus, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus, Geobacillus and probacteria such as Lactobacillus. 23. The encapsulated probiotic cells according to claim 18, wherein the probiotic cells are selected from the group comprising Saccharomyces cereviseae, Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, Bifidobacterium angulatum, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium lactis, Bifidobacterium longum, Enterococcus faecium, Enterococcus faecalis, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus alimentarius, Lactobacillus bulgaricus, Lactobacillus casei subsp. casei, Lactobacillus casei Shirota, Lactobacillus curvatus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus fermentum, Lactobacillus farciminus, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus lacti, Lactobacillus paracasei, Lactobacillus pentosaceus, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus (Lactobacillus GG), Lactobacillus sake, Lactobacillus salivarius, Lactococcus lactis, Micrococcus varians, Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus acidilactici, Pediococcus halophilus, Streptococcus faecalis, Streptococcus thermophilus, Staphylococcus carnosus, and Staphylococcus xylosus. 24. (canceled) 25. (canceled) 26. (canceled) 27. Encapsulated Lactobacillus acidophilus or Bacillus subtilis cells, wherein the encapsulated cells produce and excrete digestive enzymes and wherein the cells are encapsulated in a microcapsule having a porous capsule wall, wherein the porous capsule wall comprises a complex formed from sodium cellulose sulphate and poly[dimethyldiallyl-ammonium chloride], thereby providing that these encapsulated cells are resistant to a treatment with acidic aqueous solution of a pH value of 2 for a time period of 2 to 4 hours. 28. The encapsulated cells according to claim 2, wherein the digestive enzymes are selected from the group comprising alpha amylases, glucoamylases, alpha galactosidases, proteases, bromelain proteases, subtilisin, cellulases, pectinases and lipases. 29. (canceled) 30. The encapsulated microbial cells according to claim 1 formulated as a food supplement comprising an excipient acceptable for food use. 31. The encapsulated microbial cells according to claim 1 formulated as a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a suitable carrier. 32. A method to protect microbial cells from being degraded by treatment with an acidic aqueous solution by microencapsulation comprising
a) suspending the living microbial cells in an aqueous solution of a polyelectrolyte sodium cellulose sulphate b) introducing the suspension in form of preformed microcapsules into a precipitation bath containing an aqueous solution of the counter-charged polyelectrolyte poly[dimethyldiallyl-ammonium chloride], c) terminating the reaction in the bath after 1-10 minutes, preferably 3-5 minutes, and more preferably after 4 minutes, d) harvesting the encapsulated cells from the bath. 33. (canceled) 34. (canceled) 35. (canceled) 36. (canceled) | A simple cellulose sulphate based microencapsulation technology has been applied to encapsulate bacterial or other microbial cells, which produce and release digestive enzymes and thereby provides an acid resistant shelter for these microbial cells. Surprisingly, the resulting spheres were found to provide sufficient protection for encapsulated cells from treatment with aqueous acidic solutions. Thereby the cellulose sulphate microencapsulated cells, such as probiotics are now enabled to survive passage, for example, through the stomach after consumption by a human or animal with a higher survival rate than those not within a microcapsule. After passing the stomach these cells are delivering products produced by them, e.g. enzymes or other nutrition factors. This technology therefore proves to be very useful in providing digestive or otherwise beneficial enzymes and/or of living microbial cells, into the lower gastrointestinal tract, where they could confer their health benefit to the host.1. Encapsulated microbial cells, wherein the microbial cells are encapsulated in a microcapsule having a porous capsule wall, wherein the porous capsule wall comprises a complex formed from sodium cellulose sulphate and poly[dimethyldiallyl-ammonium chloride], and wherein the microcapsule protects the microbial cells from being degraded by acidic aqueous solution. 2. The encapsulated microbial cells according to claim 1 wherein the cells produce and excrete digestive enzymes, and wherein the porous capsule wall is permeable to said digestive enzymes. 3. The encapsulated microbial cells according to claim 1, wherein the acidic aqueous solution is simulated gastric juice, gastric juice or gastric acid. 4. The encapsulated microbial cells according to claim 2, wherein the microcapsules release the enzymes generated by the microbial cells upon treatment with intestinal fluid or duodenal fluid. 5. The encapsulated microbial cells according to claim 4, wherein the intestinal fluid is simulated intestinal fluid (SIF) and the duodenal fluid is simulated duodenal fluid (SDF). 6. The encapsulated microbial cells according to claim 1, wherein the majority of encapsulated microbial cells survive a treatment with acidic aqueous solution according to claim 3 having a pH range between 1.0 and 3.0, preferably between 1.5 and 2.5, most preferably of 2.0 for at least 1.5 hours, preferably for at least 2.5, and more preferably for at least 4 hours. 7. The encapsulated microbial cells according to claim 6, wherein the majority of microbial cells survive a passage through the stomach of an animal. 8. The encapsulated microbial cells according to claim 7, wherein the majority is defined as a value selected from the group consisting of at least 51% of the cells, as 60% to 90% of the cells, as 60% to 80% of the cells and as 60% of the cells. 9. (canceled) 10. (canceled) 11. (canceled) 12. The encapsulated microbial cells according to claim 1, wherein the microbial cells are at least partially released in the gut of an animal. 13. The encapsulated microbial cells according to claim 1, wherein the animal is a mammal or an avian. 14. (canceled) 15. (canceled) 16. The encapsulated microbial cells according to claim 1 wherein the microcapsules have a diameter of between 0.01 and 5 mm, preferably between 0.05 and 3 mm, and most preferably between 0.1 and 1 mm. 17. The encapsulated microbial cells according to claim 1, wherein the surface pores of the porous capsule wall have a molecular weight cut off between 50 and 200 kDa, preferably between 60-150 kDa and most preferably between 60 and 100 kDa. 18. The encapsulated microbial cells according to claim 1, wherein the microbial cells are selected from the group comprising bacterial cells, yeast cells, fungal cells and probiotic cells. 19. (canceled) 20. (canceled) 21. (canceled) 22. The encapsulated bacterial cells according to claim 18, wherein the bacterial cells are selected from the group comprising Bifidobacterium, Bacteroides, Clostridium, Fusobacterium, Melissococcus, Propionibacterium, Streptococcus, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus, Geobacillus and probacteria such as Lactobacillus. 23. The encapsulated probiotic cells according to claim 18, wherein the probiotic cells are selected from the group comprising Saccharomyces cereviseae, Bacillus coagulans, Bacillus licheniformis, Bacillus subtilis, Bifidobacterium angulatum, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium lactis, Bifidobacterium longum, Enterococcus faecium, Enterococcus faecalis, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus alimentarius, Lactobacillus bulgaricus, Lactobacillus casei subsp. casei, Lactobacillus casei Shirota, Lactobacillus curvatus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus fermentum, Lactobacillus farciminus, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus lacti, Lactobacillus paracasei, Lactobacillus pentosaceus, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus (Lactobacillus GG), Lactobacillus sake, Lactobacillus salivarius, Lactococcus lactis, Micrococcus varians, Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus acidilactici, Pediococcus halophilus, Streptococcus faecalis, Streptococcus thermophilus, Staphylococcus carnosus, and Staphylococcus xylosus. 24. (canceled) 25. (canceled) 26. (canceled) 27. Encapsulated Lactobacillus acidophilus or Bacillus subtilis cells, wherein the encapsulated cells produce and excrete digestive enzymes and wherein the cells are encapsulated in a microcapsule having a porous capsule wall, wherein the porous capsule wall comprises a complex formed from sodium cellulose sulphate and poly[dimethyldiallyl-ammonium chloride], thereby providing that these encapsulated cells are resistant to a treatment with acidic aqueous solution of a pH value of 2 for a time period of 2 to 4 hours. 28. The encapsulated cells according to claim 2, wherein the digestive enzymes are selected from the group comprising alpha amylases, glucoamylases, alpha galactosidases, proteases, bromelain proteases, subtilisin, cellulases, pectinases and lipases. 29. (canceled) 30. The encapsulated microbial cells according to claim 1 formulated as a food supplement comprising an excipient acceptable for food use. 31. The encapsulated microbial cells according to claim 1 formulated as a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a suitable carrier. 32. A method to protect microbial cells from being degraded by treatment with an acidic aqueous solution by microencapsulation comprising
a) suspending the living microbial cells in an aqueous solution of a polyelectrolyte sodium cellulose sulphate b) introducing the suspension in form of preformed microcapsules into a precipitation bath containing an aqueous solution of the counter-charged polyelectrolyte poly[dimethyldiallyl-ammonium chloride], c) terminating the reaction in the bath after 1-10 minutes, preferably 3-5 minutes, and more preferably after 4 minutes, d) harvesting the encapsulated cells from the bath. 33. (canceled) 34. (canceled) 35. (canceled) 36. (canceled) | 1,600 |
1,051 | 15,500,418 | 1,634 | The present invention relates to a method for selecting a target region of interest (ROI) in a target nucleic acid molecule using a nucleic acid probe comprising a 3′ sequence capable of hybridising to a target nucleic acid molecule and acting as a primer for the production of a complement of the target ROI (i.e. by target templated extension of the primer), and a sequence capable of templating the circularisation and ligation of the extended probe comprising the reverse complement of the target ROI and a portion of the probe. The circularised molecule thus obtained contains the reverse complement of the target ROI and may be subjected to further analysis and/or amplification etc. The probe may be provided as an oligonucleotide comprising a stem-loop structure or as a partially double-stranded construct and comprises a single-stranded 3′ end region containing the target-binding site. A second binding site provided in the probe serves as the ligation template for circularisation, and the stem-loop structure, if present, is cleaved to render the second binding site available for hybridisation to the target complement. Also provided are probes and kits for carrying out such a method. | 1. A method of selecting a target region of interest (ROI) in a target nucleic acid molecule, said ROI being flanked by a 3′ flanking sequence and a 5′ flanking sequence in the target molecule, said method comprising:
(a) providing a probe comprising
(i) a first target-binding site at a 3′ end region of said probe, which binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being the 3′ flanking sequence flanking the 3′ end of the ROI, and is capable of being extended to generate a complement of the target molecule in a target-templated extension reaction, said target complement comprising the complement of said 3′ flanking sequence and at least of the ROI and the 5′ flanking sequence; and
(ii) a second binding site which is homologous to the 5′ flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence in said target complement;
wherein said probe is provided as an oligonucleotide comprising a stem-loop structure which comprises the second binding site in the loop of the structure and further comprises a cleavage site 3′ of the second binding site which is cleavable to open the loop to render the second binding site available for binding, and a single-stranded region at the 3′ end comprising the first binding site; or
wherein said probe is provided as a partially double-stranded construct comprising a first strand comprising a single-stranded 3′ end region comprising the first binding site at the 3′ end thereof and a second strand hybridised at the 5′ end of said first strand and comprising a single-stranded 3′ end region comprising the second binding site;
(b) contacting the probe with the target molecule and allowing the first target-binding site at the 3′ end of the probe to hybridise to its complementary binding site in the target molecule, wherein the target molecule is at least partially single stranded including in the region comprising the complementary binding site, and optionally the ROI and the 5′ flanking sequence;
(c) extending the hybridised 3′ end of the probe using the target molecule as an extension template to generate a complement of the target molecule;
(d) removing the target molecule, leaving an extended probe comprising 3′ to 5′ in the extended region complements of the 5′ flanking sequence, the ROI and the 3′ flanking sequence of the target molecule, wherein the second binding site remains in the probe;
(e) allowing the extended probe to undergo an intramolecular rearrangement such that the second binding site hybridises to its complementary binding site in the target complement, being the complement of the 5′ flanking sequence of the target molecule,
wherein if said probe comprises a stem-loop structure the rearrangement comprises cleavage of the extended probe at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a second strand hybridised to the first strand in the stem and comprising the released 3′ end which is then able to hybridise to its complementary binding site in the first strand, and
wherein if said 5′ flanking sequence is internal to the 5′ end of the target molecule and the target complement in the first extended strand contains an additional sequence 3′ of the complement of the 5′ flanking sequence, the rearrangement comprises a cleavage in or of the additional sequence to release the 3′ end of the first strand comprising the complementary binding site for the second binding site,
such that the, optionally released, 5′ end of the first, extended, strand and the, optionally released, 3′ end of the first, extended, strand are brought into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template;
(f) ligating the ends of the extended strand of the probe directly or indirectly to one another to circularise the extended strand of the probe;
(g) amplifying or separating the circularised extended strand, thereby to select the ROI. 2. The method of claim 1, being a method of selecting a target ROI in a target nucleic acid molecule, said method comprising;
(a) providing a probe comprising a stem-loop structure and a single-stranded region at the 3′ end, wherein said 3′ end region comprises a first target-binding site which is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a flanking sequence flanking the 3′ end of the ROI, and said loop comprises a second binding site which is homologous to a flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence, and wherein said stem-loop structure further comprises a cleavage site 3′ of said second binding site such that cleavage allows the loop to open; (b) contacting the probe with the target molecule and allowing the first target-binding site at the 3′ end of the probe to hybridise to its complementary binding site in the target molecule, wherein the target molecule is at least partially single stranded including in the region comprising the complementary binding site, and optionally the ROI and the 5′ flanking sequence; (c) extending the hybridised 3′ end of the probe using the target molecule as an extension template to generate a complement of the target molecule; (d) removing the target molecule, leaving an extended probe comprising 3′ to 5′ in the extended region complements of the 5′ flanking sequence and the ROI, and the 3′ flanking sequence of the target molecule; (e) allowing the extended probe to undergo an intramolecular rearrangement such that the second binding site is able to hybridise to its complementary binding site in the target complement, being the complement of the 5′ flanking sequence of the target molecule, wherein the rearrangement comprises cleavage of the extended probe at least at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a released 5′ end, and a second strand hybridised to the first strand in the stem and comprising the released 3′ end which is then able to hybridise to its complementary binding site in the first strand, and if said 5′ flanking sequence is internal to the 5′ end of the target molecule and the target complement in the first extended strand contains an additional sequence 3′ of the complement of the 5′ flanking sequence, also a second cleavage in or of the additional sequence to release the 3′ end of the first strand comprising the complementary binding site for the second binding site, such that the released 5′ end of the first, extended, strand and the optionally released 3′ end of the first, extended, strand are brought into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template; (f) ligating the ends of the extended strand of the probe directly or indirectly to one another to circularise the extended strand of the probe; (g) amplifying or separating the circularised extended strand, thereby to select the ROI. 3. The method of claim 2, wherein the 5′ flanking sequence lies at the 5′ end of the target nucleic acid molecule and step (e) comprises
(i) cleaving the extended probe at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site, thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a second strand hybridised to the first strand in the stem and comprising the released 3′ end; and
(ii) allowing the second binding site in the released 3′ end in the second strand to hybridise to its complementary binding site in the target complement in the first strand, thereby to bring the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation, directly or indirectly, to each other, using the second binding site as ligation template. 4. The method of claim 2, wherein when the 5′ flanking sequence is internal to the 5′ end of the target nucleic acid molecule, the probe further comprises a single-stranded region at the 5′ end comprising a third binding site which is homologous to a cleavage sequence immediately 5′ to the 5′ flanking sequence in the target molecule and is capable of hybridising to a complement of said cleavage sequence, wherein said third binding site is optionally separated from the 5′ end of the stem by a spacer sequence and wherein step (e) comprises:
(i) allowing the extended probe to undergo an intramolecular hybridisation wherein the third binding site hybridises to its complementary binding site in the target complement, thereby generating a second cleavage site;
(ii) cleaving the hybridised probe at the second cleavage site and at the cleavage site in the stem-loop structure, thereby generating a partially double stranded construct comprising two strands hybridised at the stem, the first strand comprising the target complement and the second strand comprising the second binding site;
(iii) allowing the second binding site to hybridise to its complementary binding site in the target complement in the first strand, thereby bringing the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template. 5. The method of claim 4, wherein the loop in the probe further comprises 5′ to the second binding site a sequence complementary to a sequence from the complementary binding site in the target complement for the third binding site, which sequence remains at the 3′ end of the first strand of the probe following cleavage at the second cleavage site. 6. The method of claim 4 or claim 5, wherein the spacer sequence comprises a fourth target binding site which is capable of hybridising to a complementary binding site in the target molecule lying 3′, preferably immediately 3′, of the 3′ flanking sequence of the target molecule, and wherein step (b) further comprises allowing the fourth target binding site to hybridise to its complementary binding site in the target molecule. 7. The method of claim 2, wherein the 5′ flanking sequence is internal to the 5′ end of the target nucleic acid molecule, and the probe further comprises within the stem-loop structure, preferably within the loop thereof, a third binding site which is homologous to a cleavage sequence immediately 5′ to the 5′ flanking sequence in the target molecule and which is capable of hybridising to a complement of said cleavage sequence, wherein said third binding site is 3′ to the second binding site and wherein step (e) comprises:
(i) allowing the extended probe to undergo an intramolecular hybridisation wherein the third binding site hybridises to its complementary binding site in the target complement, thereby generating a second cleavage site;
(ii) cleaving the hybridised probe at the second cleavage site and at the cleavage site in the stem-loop structure, thereby generating a partially double stranded construct comprising two strands hybridised at the stem, the first strand comprising the target complement and the second strand comprising the second binding site;
(iii) allowing the second binding site to hybridise to its complementary binding site in the target complement in the first strand, thereby bringing the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template. 8. The method of claim 2, wherein when the 5′ flanking sequence is internal to the 5′ end of the target molecule, step (e) comprises:
(i) cleaving the extended probe at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site, thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a second strand hybridised to the first strand in the stem and comprising the released 3′ end; and
(ii) allowing the second binding site in the released 3′ end in the second strand to hybridise to its complementary binding site in the target complement in the first strand, wherein the additional sequence at the 3′ end of the first strand does not hybridise and forms a protruding single stranded end;
(iii) cleaving the protruding single stranded end to leave a 3′ end of the first strand which is hybridised to the second binding site in the second strand, thereby to bring the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation, directly or indirectly, to each other, using the second binding site as ligation template. 9. The method of claim 1, being a method of selecting a target region of interest (ROI) in a target nucleic acid molecule, said method comprising;
(a) providing a probe being a partially double-stranded construct having a first strand comprising single-stranded 3′ end region comprising a first target-binding site at the 3′ end thereof, wherein said first target binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a flanking sequence flanking the 3′ end of the ROI, and a second strand hybridised to said first strand at the 5′ end thereof and comprising a single-stranded 3′ end region comprising a second binding site, wherein the second binding site is homologous to a flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence; (b) contacting the probe with the target molecule and allowing the first target-binding site at the 3′ end of the first strand of the probe to hybridise to its complementary binding site in the target molecule, wherein the target molecule is at least partially single stranded including in the region comprising the complementary binding site, and optionally the ROI and the 5′ flanking sequence; (c) extending the hybridised 3′ end of the first strand of the probe using the target molecule as an extension template to generate a complement of the target molecule; (d) removing the target molecule without denaturing the probe, leaving an extended probe comprising 3′ to 5′ in the extended region complements of the 5′ flanking sequence, the ROI and the 3′ flanking sequence of the target molecule; (e) allowing the extended probe to undergo an intramolecular rearrangement such that the second binding site in the second strand is able to hybridise to its complementary binding site in the target complement in the extended first strand, being the complement of the 5′ flanking sequence of the target molecule, wherein if said 5′ flanking sequence is internal to the 5′ end of the target molecule and the target complement in the first extended strand contains an additional sequence 3′ of the complement of the 5′ flanking sequence, the rearrangement comprises a cleavage in or of the additional sequence to release the 3′ end of the first strand comprising the complementary binding site for the second binding site,
such that the 5′ end of the first, extended, strand and the optionally released 3′ end of the first, extended, strand are brought into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template;
(f) ligating the ends of the extended strand of the probe directly or indirectly to one another to circularise the extended strand of the probe; (g) amplifying or separating the circularised extended strand, thereby to select the ROI. 10. The method of claim 9, wherein the 5′ flanking sequence lies at the 5′ end of the target nucleic acid molecule and step (e) comprises:
allowing the second binding site in the second strand to hybridise to its complementary binding site in the target complement in the extended first strand, thereby to bring the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation, directly or indirectly to each other, using the second binding site as ligation template. 11. The method of claim 1 or 9, wherein the 5′ flanking sequence is internal to the 5′ end of the target nucleic acid molecule, and the second strand of the probe further comprises a third binding site which is homologous to a cleavage sequence immediately 5′ to the 5′ flanking sequence in the target molecule and is capable of hybridising to a complement of said cleavage sequence in the additional sequence in the target complement in the extended first strand which is 3′ to the complement of the 5′ flanking sequence, wherein said third binding site is located in the single-stranded 3′ end region of the second strand 3′ to the second binding site, or in a further single-stranded 5′ end region of the second strand;
and wherein step (e) comprises:
(i) allowing the extended probe to undergo an intramolecular hybridisation wherein the third binding site hybridises to its complementary binding site in the target complement, thereby generating a cleavage site;
(ii) cleaving the hybridised probe at the cleavage site, thereby generating a released 3′ end of the first extended strand;
(iii) allowing the second binding site to hybridise to its complementary binding site in the target complement in the first strand, thereby bringing the 5′ end and the released 3′ end of the first, extended, strand into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template. 12. The method of claim 3 or 10 wherein the target molecule is a micro RNA or a RNA transcript. 13. The method of any one of claims 1 to 12, wherein the probe comprises a further probe sequence which lies 3′ of the second binding site and is homologous to the 3′ flanking sequence and complementary to the first binding site, wherein when said probe comprises a stem-loop structure said further probe sequence is located in the loop or when said probe is a double-stranded construct the further probe sequence is located at the 3′ end of the second strand; and wherein in step (e) of said method the further probe sequence hybridises to the first binding site and acts as ligation template together with the second binding site. 14. A method of selecting a target ROI in a target nucleic acid molecule, said method comprising:
(a) providing a probe comprising
(i) a first target-binding site at a 3′ end region of said probe, which binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a 3′ flanking sequence flanking the 3′ end of the ROI, and is capable of being extended to generate a complement of the target molecule in a target-templated extension reaction, said target complement comprising the complement of said 3′ flanking sequence and the ROI; and
(ii) a second binding site which is homologous to the ROI, or to the 5′ end thereof and is capable of hybridising to a complement of the ROI, or of the 5′ end thereof, in said target complement;
(iii) a further probe sequence, lying 3′ of the second binding site, which is homologous to the 3′ flanking sequence and complementary to the first binding site;
wherein said probe is provided as an oligonucleotide comprising a stem-loop structure which comprises the second binding site and the further probe sequence in the loop of the structure and further comprises a cleavage site 3′ of the second binding site and further probe sequence, which is cleavable to open the loop to render the second binding site and further probe sequence available for binding, and a single-stranded region at the 3′ end of the probe comprising the first binding site at the 3′ end thereof; or
wherein said probe is provided as a partially double-stranded construct comprising a first strand comprising a single-stranded 3′ end region comprising the first binding site at the 3′ end thereof and a second strand hybridised at the 5′ end of said first strand and comprising a single-stranded 3′ end region comprising the second binding site and the further probe sequence at the 3′ end of the single-stranded region;
(b) contacting the probe with the target molecule and allowing the first target-binding site at the 3′ end of the probe to hybridise to its complementary binding site in the target molecule, wherein the target molecule is at least partially single stranded including in the region comprising the complementary binding site, and optionally the ROI; (c) extending the hybridised 3′ end of the probe using the target molecule as an extension template to generate a complement of the target molecule; (d) removing the target molecule, leaving an extended probe comprising 3′ to 5′ in the extended region a complement of the ROI, and the 3′ flanking sequence of the target molecule, wherein the second binding site remains in the probe; (e) allowing the extended probe to undergo an intramolecular rearrangement such that the second binding site hybridises to its complementary binding site in the target complement, being the complement of the ROI, or of the 5′ end of the ROI, and the further probe sequence hybridises to the first binding site,
wherein if said probe comprises a stem-loop structure the rearrangement comprises cleavage of the extended probe at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site and the further probe sequence, thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a released 5′ end, and a second strand hybridised to the first strand in the stem and comprising the released 3′ end which is then able to hybridise to its complementary binding sites in the first strand, and
wherein if said ROI is internal to the 5′ end of the target molecule and the target complement in the first extended strand contains an additional sequence 3′ of the complement of the ROI, the rearrangement comprises a cleavage in or of the additional sequence to release the 3′ end of the first strand comprising the complementary binding site for the second binding site.
such that the, optionally released, 5′ end of the first, extended, strand and the, optionally released, 3′ end of the first, extended, strand are brought into juxtaposition for ligation directly or indirectly to each other, using the second binding site and further probe sequence as ligation template;
(f) ligating the ends of the extended strand of the probe directly or indirectly to one another to circularise the extended strand of the probe; (g) amplifying or separating the circularised extended strand, thereby to select the ROI. 15. The method of any one of claim 4 to 6, or 11, wherein the probe comprises an immobilisable affinity binding group or capture sequence element at the 5′ end of a stem-loop probe or at the 5′ end of the second strand of a partially-stranded probe, wherein said imobilisable group or sequence element is located at the 5′ end of a 5′ single-stranded region of the probe comprising a third binding site, and wherein said immobilisable group or sequence element is cleaved from extended probes upon cleavage at the cleavage site created by the third binding site allowing unreacted probes which have not been extended and cleaved at the third binding site and which retain the immobilisable group or sequence element to be removed or separated by virtue of the immobilisable group or sequence element. 16. The method of any one of claims 5 to 15, wherein the probe comprises a stem loop structure and wherein the cleavage site is in the loop of the probe, preferably wherein the cleavage site in the loop is adjacent to the stem. 17. The method of any one of claims 5 to 15, wherein the probe comprises a stem loop structure and wherein the cleavage site is in the stem of the probe, preferably wherein the cleavage site in the stem is adjacent to the loop. 18. The method of any one of claims 5 to 17, wherein the stem or double-stranded region of the probe comprises one or more tag sequences. 19. The method of any one of claims 1 to 18, wherein the target molecule is RNA and it is removed in step (d) by RNase digestion or alkaline hydrolysis. 20. The method of any one of claims 5 to 18, wherein the probe comprises a stem loop structure and wherein the target molecule is removed in step (d) by denaturation. 21. The method of any one of claims 1 to 20, wherein a plurality of probes are used to select a plurality of target regions of interest within the same target molecule derived from a variety of sources. 22. The method of any one of claims 1 to 21, wherein a single probe is used to select a plurality of different target regions of interest within the same target molecule derived from a variety of sources. 23. The method of any one of claims 1 to 22, wherein the probe comprises one or more spacer sequences. 24. The method of claim 23, wherein
a) the probe comprises a stem loop structure and one or more spacer sequences is located between the first target binding site and the stem-loop structure, within the stem-loop structure, 5′ of the stem loop structure, in the stem and/or in the loop; or b) the probe is a partially double-stranded structure and comprises one or more spacer sequences in the first strand between the first target binding site and the double-stranded region or 5′ to the double-stranded region, or in the second strand 5′ to the double-stranded region or 3′ to the double-stranded region. 25. The method of claim 23 or 24, wherein the probe comprises a third binding site, and further comprises one or more spacer sequences 3′ or 5′ to said third binding site, preferably wherein said one or more spacer sequences are directly adjacent to said third binding site. 26. The method of claim 23, wherein
a) the probe comprises a stem loop structure and the one or more spacer sequences is located 3′ of the stem of the stem-loop structure; or b) the probe is a partially double-stranded structure and comprises one or more spacer sequences in the first strand 3′ of the double-stranded region. 27. The method of any one of claims 23 to 26, wherein the spacer sequence is a capture sequence. 28. The method of claim 27, wherein the capture sequence hybridises to a cognate complementary binding site provided on a solid surface. 29. The method of claim 27, wherein the capture sequence or a complementary oligonucleotide hybridised thereto is attached to an affinity binding moiety. 30. The method of any one of claims 27 to 29, wherein
a) the probe comprises a stem loop structure and a capture sequence is provided 5′ of the stem of the stem-loop structure; or
b) the probe is a partially double-stranded structure and comprises a capture sequence in the second strand 5′ of the double-stranded region. 31. The method of any one of claims 23 to 30, wherein the spacer sequence is or comprises a tag sequence or a complement thereof, wherein the tag sequence is selected from a detection sequence or an identification sequence element or a binding site for a primer or detection probe. 32. The method of claim 31, wherein the tag sequence is an identification element for the target ROI or a sample identification sequence. 33. The method of any one of claims 23 to 26, wherein the probe comprises a stem loop structure and the spacer sequence is located within the loop of the stem-loop structure and is located 5′ of the second binding site thereby creating a gap between the respective ends of the extended strand when they are both hybridised to the second binding site in step (e), and wherein the gap is filled prior to ligation by one or more gap oligonucleotides which hybridise in the gap between the ends of the extended strand, or by gap-fill extension of the 3′ end of the hybridised extended strand using a polymerase, such that the extended strand and any gap oligonucleotides if present may be ligated into a circular molecule comprising the complement of the target fragment. 34. The method of claim 33, wherein the gap oligonucleotide(s) comprise(s) a tag sequence complementary to a tag sequence complement in the spacer sequence. 35. The method of claim 33, wherein the gap oligonucleotide comprises a region which is not complementary to the spacer sequence, and wherein the region of non-complementarity comprises a tag sequence. 36. The method of any one of claims 33 to 35, wherein the gap oligonucleotide comprises a detection sequence or an identification sequence element, or a binding site for an amplification primer or a detection probe. 37. The method of claim 36, wherein the amplification primer is a universal amplification primer. 38. The method of any one of claims 33 to 37, which is performed in multiplex using a plurality of different probes and wherein the gap oligonucleotide for each probe comprises the same tag sequence, preferably the same primer binding site. 39. The method of any one of claims 33 to 38, wherein the gap oligonucleotide is pre-hybridised to the probe prior to contacting the probe with the target nucleic acid molecule. 40. The method of any one of claims 33 to 38, wherein the gap oligonucleotide is separately provided at the same time or after contacting the probe with the target molecule. 41. The method of any one of claims 1 to 40, wherein the probe comprises a stem loop structure and the loop of the stem-loop structure comprises a region of intramolecular complementarity such that it is able to form a duplex within the loop, preferably wherein the duplex within the loop comprises a cleavage site. 42. The method of any one of claims 1 to 9 or 11 to 41, wherein cleavage is enzymatic cleavage. 43. The method of any one of claims 1 to 9 or 11 to 42, wherein the cleavage site is recognised by one or more enzymes capable of cleaving nucleic acid molecules. 44. The method of claim 43, wherein said one or more enzymes is a nickase or a restriction endonuclease. 45. The method of claim 44, wherein the nickase enzyme is removed from the assay or inactivated following unfolding of the probe. 46. The method of claim 42, wherein the enzymatic cleavage comprises a uracil-DNA glycosylase (UNG) enzyme in combination with an endonuclease enzyme capable of recognising apurinic/apyrimidinic sites of dsDNA. 47. The method of claim 46, wherein the endonuclease enzyme is capable of recognising apurinic/apyrimidinic sites of dsDNA is endonuclease IV. 48. The method of any one of claims 1 to 47, wherein amplification of the circularised extended strand of the probe is by PCR or Rolling Circle Amplification (RCA). 49. The method of claim 48, wherein amplification is by RCA, and wherein amplification further comprises a second round of RCA. 50. The method of any one of claims 1 to 49 further comprising a step of detecting and/or analysing the circularised extended strand or an amplicon thereof. 51. The method of claim 50, wherein the amplified product is detected by hybridising a detection probe labelled with a directly or indirectly detectable label to the amplification product. 52. The method of claim 51, wherein the detection label is a fluorescent label. 53. The method of claim 51, wherein the extended strand or amplicon thereof is detected or analysed by sequencing. 54. A probe for use in the method of any one of claims 1 to 8, or 11 to 53, said probe comprising a stem-loop structure and a single-stranded region at the 3′ end, wherein said 3′ end region comprises a first target-binding site which is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a flanking sequence flanking the 3′ end of the ROI, and said loop comprises a second binding site which is homologous to a flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence, and wherein said stem-loop structure further comprises a cleavage site 3′ of said second binding site such that cleavage allows the loop to open. 55. A probe for use in the method of any one of claims 1 or 9 to 53, said probe being a partially double-stranded construct having a first strand comprising single-stranded 3′ end region comprising a first target-binding site at the 3′ end thereof, wherein said first target binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a flanking sequence flanking the 3′ end of the ROI, and a second strand hybridised to said first strand at the 5′ end thereof and comprising a single-stranded 3′ end region comprising a second binding site, wherein the second binding site is homologous to a flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence. 56. A probe for use in the method of any one of claims 14 to 53, said probe comprising
(i) a first target-binding site at a 3′ end region of said probe, which binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a 3′ flanking sequence flanking the 3′ end of the ROI, and is capable of being extended to generate a complement of the target molecule in a target-templated extension reaction, said target complement comprising the complement of said 3′ flanking sequence and the ROI; and
(ii) a second binding site which is homologous to the ROI or to the 5′ end thereof and is capable of hybridising to a complement of the ROI or the 5′ end thereof;
(iii) a further probe sequence, lying 3′ of the second binding site, which is homologous to the 3′ flanking sequence and complementary to the first binding site;
wherein said probe is provided as an oligonucleotide comprising a stem-loop structure which comprises the second binding site and the further probe sequence in the loop of the structure and further comprises a cleavage site 3′ of the second binding site and further probe sequence, which is cleavable to open the loop to render the second binding site and further probe sequence available for binding, and a single-stranded region at the 3′ end of the probe comprising the first binding site at the 3′ end thereof; or
wherein said probe is provided as a partially double-stranded construct comprising a first strand comprising a single-stranded 3′ end region comprising the first binding site at the 3′ end thereof and a second strand hybridised at the 5′ end of said first strand and comprising a single-stranded 3′ end region comprising the second binding site and the further probe sequence at the 3′ end of the single-stranded region. 57. The probe of claim 54, wherein the probe is as defined in any one of claim 4 to 7, 15 to 18, 23 to 33 or 41. 58. The probe of claim 55, wherein the probe is as defined in any one of claims 11, 13, 15 or 23 to 32. 59. The probe of claim 56, wherein the probe is as defined in any one of claim 16 to 18, 23 to 33 or 41. 60. A kit for selecting a target region of interest in a target nucleic acid molecule, said kit comprising:
(a) a probe as defined in any one of claims 54 to 59; and optionally one or more further components selected from: (b) means for cleaving the cleavage site within the stem-loop structure of the probe; (c) means for cleaving the second cleavage site; (d) means for degrading the 3′ end of the extended probe, e.g. when the 5′ flanking sequence is internal to the 5′ end of the target molecule; (e) means for extending the probe, e.g. a polymerase enzyme; (f) a ligase enzyme; (g) one or more gap oligonucleotides; (h) means for amplification of the circularised extended strand;
(i) means for detecting the circularised extended strand or an amplicon thereof. | The present invention relates to a method for selecting a target region of interest (ROI) in a target nucleic acid molecule using a nucleic acid probe comprising a 3′ sequence capable of hybridising to a target nucleic acid molecule and acting as a primer for the production of a complement of the target ROI (i.e. by target templated extension of the primer), and a sequence capable of templating the circularisation and ligation of the extended probe comprising the reverse complement of the target ROI and a portion of the probe. The circularised molecule thus obtained contains the reverse complement of the target ROI and may be subjected to further analysis and/or amplification etc. The probe may be provided as an oligonucleotide comprising a stem-loop structure or as a partially double-stranded construct and comprises a single-stranded 3′ end region containing the target-binding site. A second binding site provided in the probe serves as the ligation template for circularisation, and the stem-loop structure, if present, is cleaved to render the second binding site available for hybridisation to the target complement. Also provided are probes and kits for carrying out such a method.1. A method of selecting a target region of interest (ROI) in a target nucleic acid molecule, said ROI being flanked by a 3′ flanking sequence and a 5′ flanking sequence in the target molecule, said method comprising:
(a) providing a probe comprising
(i) a first target-binding site at a 3′ end region of said probe, which binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being the 3′ flanking sequence flanking the 3′ end of the ROI, and is capable of being extended to generate a complement of the target molecule in a target-templated extension reaction, said target complement comprising the complement of said 3′ flanking sequence and at least of the ROI and the 5′ flanking sequence; and
(ii) a second binding site which is homologous to the 5′ flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence in said target complement;
wherein said probe is provided as an oligonucleotide comprising a stem-loop structure which comprises the second binding site in the loop of the structure and further comprises a cleavage site 3′ of the second binding site which is cleavable to open the loop to render the second binding site available for binding, and a single-stranded region at the 3′ end comprising the first binding site; or
wherein said probe is provided as a partially double-stranded construct comprising a first strand comprising a single-stranded 3′ end region comprising the first binding site at the 3′ end thereof and a second strand hybridised at the 5′ end of said first strand and comprising a single-stranded 3′ end region comprising the second binding site;
(b) contacting the probe with the target molecule and allowing the first target-binding site at the 3′ end of the probe to hybridise to its complementary binding site in the target molecule, wherein the target molecule is at least partially single stranded including in the region comprising the complementary binding site, and optionally the ROI and the 5′ flanking sequence;
(c) extending the hybridised 3′ end of the probe using the target molecule as an extension template to generate a complement of the target molecule;
(d) removing the target molecule, leaving an extended probe comprising 3′ to 5′ in the extended region complements of the 5′ flanking sequence, the ROI and the 3′ flanking sequence of the target molecule, wherein the second binding site remains in the probe;
(e) allowing the extended probe to undergo an intramolecular rearrangement such that the second binding site hybridises to its complementary binding site in the target complement, being the complement of the 5′ flanking sequence of the target molecule,
wherein if said probe comprises a stem-loop structure the rearrangement comprises cleavage of the extended probe at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a second strand hybridised to the first strand in the stem and comprising the released 3′ end which is then able to hybridise to its complementary binding site in the first strand, and
wherein if said 5′ flanking sequence is internal to the 5′ end of the target molecule and the target complement in the first extended strand contains an additional sequence 3′ of the complement of the 5′ flanking sequence, the rearrangement comprises a cleavage in or of the additional sequence to release the 3′ end of the first strand comprising the complementary binding site for the second binding site,
such that the, optionally released, 5′ end of the first, extended, strand and the, optionally released, 3′ end of the first, extended, strand are brought into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template;
(f) ligating the ends of the extended strand of the probe directly or indirectly to one another to circularise the extended strand of the probe;
(g) amplifying or separating the circularised extended strand, thereby to select the ROI. 2. The method of claim 1, being a method of selecting a target ROI in a target nucleic acid molecule, said method comprising;
(a) providing a probe comprising a stem-loop structure and a single-stranded region at the 3′ end, wherein said 3′ end region comprises a first target-binding site which is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a flanking sequence flanking the 3′ end of the ROI, and said loop comprises a second binding site which is homologous to a flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence, and wherein said stem-loop structure further comprises a cleavage site 3′ of said second binding site such that cleavage allows the loop to open; (b) contacting the probe with the target molecule and allowing the first target-binding site at the 3′ end of the probe to hybridise to its complementary binding site in the target molecule, wherein the target molecule is at least partially single stranded including in the region comprising the complementary binding site, and optionally the ROI and the 5′ flanking sequence; (c) extending the hybridised 3′ end of the probe using the target molecule as an extension template to generate a complement of the target molecule; (d) removing the target molecule, leaving an extended probe comprising 3′ to 5′ in the extended region complements of the 5′ flanking sequence and the ROI, and the 3′ flanking sequence of the target molecule; (e) allowing the extended probe to undergo an intramolecular rearrangement such that the second binding site is able to hybridise to its complementary binding site in the target complement, being the complement of the 5′ flanking sequence of the target molecule, wherein the rearrangement comprises cleavage of the extended probe at least at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a released 5′ end, and a second strand hybridised to the first strand in the stem and comprising the released 3′ end which is then able to hybridise to its complementary binding site in the first strand, and if said 5′ flanking sequence is internal to the 5′ end of the target molecule and the target complement in the first extended strand contains an additional sequence 3′ of the complement of the 5′ flanking sequence, also a second cleavage in or of the additional sequence to release the 3′ end of the first strand comprising the complementary binding site for the second binding site, such that the released 5′ end of the first, extended, strand and the optionally released 3′ end of the first, extended, strand are brought into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template; (f) ligating the ends of the extended strand of the probe directly or indirectly to one another to circularise the extended strand of the probe; (g) amplifying or separating the circularised extended strand, thereby to select the ROI. 3. The method of claim 2, wherein the 5′ flanking sequence lies at the 5′ end of the target nucleic acid molecule and step (e) comprises
(i) cleaving the extended probe at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site, thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a second strand hybridised to the first strand in the stem and comprising the released 3′ end; and
(ii) allowing the second binding site in the released 3′ end in the second strand to hybridise to its complementary binding site in the target complement in the first strand, thereby to bring the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation, directly or indirectly, to each other, using the second binding site as ligation template. 4. The method of claim 2, wherein when the 5′ flanking sequence is internal to the 5′ end of the target nucleic acid molecule, the probe further comprises a single-stranded region at the 5′ end comprising a third binding site which is homologous to a cleavage sequence immediately 5′ to the 5′ flanking sequence in the target molecule and is capable of hybridising to a complement of said cleavage sequence, wherein said third binding site is optionally separated from the 5′ end of the stem by a spacer sequence and wherein step (e) comprises:
(i) allowing the extended probe to undergo an intramolecular hybridisation wherein the third binding site hybridises to its complementary binding site in the target complement, thereby generating a second cleavage site;
(ii) cleaving the hybridised probe at the second cleavage site and at the cleavage site in the stem-loop structure, thereby generating a partially double stranded construct comprising two strands hybridised at the stem, the first strand comprising the target complement and the second strand comprising the second binding site;
(iii) allowing the second binding site to hybridise to its complementary binding site in the target complement in the first strand, thereby bringing the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template. 5. The method of claim 4, wherein the loop in the probe further comprises 5′ to the second binding site a sequence complementary to a sequence from the complementary binding site in the target complement for the third binding site, which sequence remains at the 3′ end of the first strand of the probe following cleavage at the second cleavage site. 6. The method of claim 4 or claim 5, wherein the spacer sequence comprises a fourth target binding site which is capable of hybridising to a complementary binding site in the target molecule lying 3′, preferably immediately 3′, of the 3′ flanking sequence of the target molecule, and wherein step (b) further comprises allowing the fourth target binding site to hybridise to its complementary binding site in the target molecule. 7. The method of claim 2, wherein the 5′ flanking sequence is internal to the 5′ end of the target nucleic acid molecule, and the probe further comprises within the stem-loop structure, preferably within the loop thereof, a third binding site which is homologous to a cleavage sequence immediately 5′ to the 5′ flanking sequence in the target molecule and which is capable of hybridising to a complement of said cleavage sequence, wherein said third binding site is 3′ to the second binding site and wherein step (e) comprises:
(i) allowing the extended probe to undergo an intramolecular hybridisation wherein the third binding site hybridises to its complementary binding site in the target complement, thereby generating a second cleavage site;
(ii) cleaving the hybridised probe at the second cleavage site and at the cleavage site in the stem-loop structure, thereby generating a partially double stranded construct comprising two strands hybridised at the stem, the first strand comprising the target complement and the second strand comprising the second binding site;
(iii) allowing the second binding site to hybridise to its complementary binding site in the target complement in the first strand, thereby bringing the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template. 8. The method of claim 2, wherein when the 5′ flanking sequence is internal to the 5′ end of the target molecule, step (e) comprises:
(i) cleaving the extended probe at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site, thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a second strand hybridised to the first strand in the stem and comprising the released 3′ end; and
(ii) allowing the second binding site in the released 3′ end in the second strand to hybridise to its complementary binding site in the target complement in the first strand, wherein the additional sequence at the 3′ end of the first strand does not hybridise and forms a protruding single stranded end;
(iii) cleaving the protruding single stranded end to leave a 3′ end of the first strand which is hybridised to the second binding site in the second strand, thereby to bring the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation, directly or indirectly, to each other, using the second binding site as ligation template. 9. The method of claim 1, being a method of selecting a target region of interest (ROI) in a target nucleic acid molecule, said method comprising;
(a) providing a probe being a partially double-stranded construct having a first strand comprising single-stranded 3′ end region comprising a first target-binding site at the 3′ end thereof, wherein said first target binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a flanking sequence flanking the 3′ end of the ROI, and a second strand hybridised to said first strand at the 5′ end thereof and comprising a single-stranded 3′ end region comprising a second binding site, wherein the second binding site is homologous to a flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence; (b) contacting the probe with the target molecule and allowing the first target-binding site at the 3′ end of the first strand of the probe to hybridise to its complementary binding site in the target molecule, wherein the target molecule is at least partially single stranded including in the region comprising the complementary binding site, and optionally the ROI and the 5′ flanking sequence; (c) extending the hybridised 3′ end of the first strand of the probe using the target molecule as an extension template to generate a complement of the target molecule; (d) removing the target molecule without denaturing the probe, leaving an extended probe comprising 3′ to 5′ in the extended region complements of the 5′ flanking sequence, the ROI and the 3′ flanking sequence of the target molecule; (e) allowing the extended probe to undergo an intramolecular rearrangement such that the second binding site in the second strand is able to hybridise to its complementary binding site in the target complement in the extended first strand, being the complement of the 5′ flanking sequence of the target molecule, wherein if said 5′ flanking sequence is internal to the 5′ end of the target molecule and the target complement in the first extended strand contains an additional sequence 3′ of the complement of the 5′ flanking sequence, the rearrangement comprises a cleavage in or of the additional sequence to release the 3′ end of the first strand comprising the complementary binding site for the second binding site,
such that the 5′ end of the first, extended, strand and the optionally released 3′ end of the first, extended, strand are brought into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template;
(f) ligating the ends of the extended strand of the probe directly or indirectly to one another to circularise the extended strand of the probe; (g) amplifying or separating the circularised extended strand, thereby to select the ROI. 10. The method of claim 9, wherein the 5′ flanking sequence lies at the 5′ end of the target nucleic acid molecule and step (e) comprises:
allowing the second binding site in the second strand to hybridise to its complementary binding site in the target complement in the extended first strand, thereby to bring the 5′ and 3′ ends of the first, extended, strand into juxtaposition for ligation, directly or indirectly to each other, using the second binding site as ligation template. 11. The method of claim 1 or 9, wherein the 5′ flanking sequence is internal to the 5′ end of the target nucleic acid molecule, and the second strand of the probe further comprises a third binding site which is homologous to a cleavage sequence immediately 5′ to the 5′ flanking sequence in the target molecule and is capable of hybridising to a complement of said cleavage sequence in the additional sequence in the target complement in the extended first strand which is 3′ to the complement of the 5′ flanking sequence, wherein said third binding site is located in the single-stranded 3′ end region of the second strand 3′ to the second binding site, or in a further single-stranded 5′ end region of the second strand;
and wherein step (e) comprises:
(i) allowing the extended probe to undergo an intramolecular hybridisation wherein the third binding site hybridises to its complementary binding site in the target complement, thereby generating a cleavage site;
(ii) cleaving the hybridised probe at the cleavage site, thereby generating a released 3′ end of the first extended strand;
(iii) allowing the second binding site to hybridise to its complementary binding site in the target complement in the first strand, thereby bringing the 5′ end and the released 3′ end of the first, extended, strand into juxtaposition for ligation directly or indirectly to each other, using the second binding site as ligation template. 12. The method of claim 3 or 10 wherein the target molecule is a micro RNA or a RNA transcript. 13. The method of any one of claims 1 to 12, wherein the probe comprises a further probe sequence which lies 3′ of the second binding site and is homologous to the 3′ flanking sequence and complementary to the first binding site, wherein when said probe comprises a stem-loop structure said further probe sequence is located in the loop or when said probe is a double-stranded construct the further probe sequence is located at the 3′ end of the second strand; and wherein in step (e) of said method the further probe sequence hybridises to the first binding site and acts as ligation template together with the second binding site. 14. A method of selecting a target ROI in a target nucleic acid molecule, said method comprising:
(a) providing a probe comprising
(i) a first target-binding site at a 3′ end region of said probe, which binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a 3′ flanking sequence flanking the 3′ end of the ROI, and is capable of being extended to generate a complement of the target molecule in a target-templated extension reaction, said target complement comprising the complement of said 3′ flanking sequence and the ROI; and
(ii) a second binding site which is homologous to the ROI, or to the 5′ end thereof and is capable of hybridising to a complement of the ROI, or of the 5′ end thereof, in said target complement;
(iii) a further probe sequence, lying 3′ of the second binding site, which is homologous to the 3′ flanking sequence and complementary to the first binding site;
wherein said probe is provided as an oligonucleotide comprising a stem-loop structure which comprises the second binding site and the further probe sequence in the loop of the structure and further comprises a cleavage site 3′ of the second binding site and further probe sequence, which is cleavable to open the loop to render the second binding site and further probe sequence available for binding, and a single-stranded region at the 3′ end of the probe comprising the first binding site at the 3′ end thereof; or
wherein said probe is provided as a partially double-stranded construct comprising a first strand comprising a single-stranded 3′ end region comprising the first binding site at the 3′ end thereof and a second strand hybridised at the 5′ end of said first strand and comprising a single-stranded 3′ end region comprising the second binding site and the further probe sequence at the 3′ end of the single-stranded region;
(b) contacting the probe with the target molecule and allowing the first target-binding site at the 3′ end of the probe to hybridise to its complementary binding site in the target molecule, wherein the target molecule is at least partially single stranded including in the region comprising the complementary binding site, and optionally the ROI; (c) extending the hybridised 3′ end of the probe using the target molecule as an extension template to generate a complement of the target molecule; (d) removing the target molecule, leaving an extended probe comprising 3′ to 5′ in the extended region a complement of the ROI, and the 3′ flanking sequence of the target molecule, wherein the second binding site remains in the probe; (e) allowing the extended probe to undergo an intramolecular rearrangement such that the second binding site hybridises to its complementary binding site in the target complement, being the complement of the ROI, or of the 5′ end of the ROI, and the further probe sequence hybridises to the first binding site,
wherein if said probe comprises a stem-loop structure the rearrangement comprises cleavage of the extended probe at the cleavage site in the stem-loop structure of the probe to release a 3′ end of the probe comprising the second binding site and the further probe sequence, thereby generating a partially double-stranded construct comprising a first extended strand comprising the target complement and a released 5′ end, and a second strand hybridised to the first strand in the stem and comprising the released 3′ end which is then able to hybridise to its complementary binding sites in the first strand, and
wherein if said ROI is internal to the 5′ end of the target molecule and the target complement in the first extended strand contains an additional sequence 3′ of the complement of the ROI, the rearrangement comprises a cleavage in or of the additional sequence to release the 3′ end of the first strand comprising the complementary binding site for the second binding site.
such that the, optionally released, 5′ end of the first, extended, strand and the, optionally released, 3′ end of the first, extended, strand are brought into juxtaposition for ligation directly or indirectly to each other, using the second binding site and further probe sequence as ligation template;
(f) ligating the ends of the extended strand of the probe directly or indirectly to one another to circularise the extended strand of the probe; (g) amplifying or separating the circularised extended strand, thereby to select the ROI. 15. The method of any one of claim 4 to 6, or 11, wherein the probe comprises an immobilisable affinity binding group or capture sequence element at the 5′ end of a stem-loop probe or at the 5′ end of the second strand of a partially-stranded probe, wherein said imobilisable group or sequence element is located at the 5′ end of a 5′ single-stranded region of the probe comprising a third binding site, and wherein said immobilisable group or sequence element is cleaved from extended probes upon cleavage at the cleavage site created by the third binding site allowing unreacted probes which have not been extended and cleaved at the third binding site and which retain the immobilisable group or sequence element to be removed or separated by virtue of the immobilisable group or sequence element. 16. The method of any one of claims 5 to 15, wherein the probe comprises a stem loop structure and wherein the cleavage site is in the loop of the probe, preferably wherein the cleavage site in the loop is adjacent to the stem. 17. The method of any one of claims 5 to 15, wherein the probe comprises a stem loop structure and wherein the cleavage site is in the stem of the probe, preferably wherein the cleavage site in the stem is adjacent to the loop. 18. The method of any one of claims 5 to 17, wherein the stem or double-stranded region of the probe comprises one or more tag sequences. 19. The method of any one of claims 1 to 18, wherein the target molecule is RNA and it is removed in step (d) by RNase digestion or alkaline hydrolysis. 20. The method of any one of claims 5 to 18, wherein the probe comprises a stem loop structure and wherein the target molecule is removed in step (d) by denaturation. 21. The method of any one of claims 1 to 20, wherein a plurality of probes are used to select a plurality of target regions of interest within the same target molecule derived from a variety of sources. 22. The method of any one of claims 1 to 21, wherein a single probe is used to select a plurality of different target regions of interest within the same target molecule derived from a variety of sources. 23. The method of any one of claims 1 to 22, wherein the probe comprises one or more spacer sequences. 24. The method of claim 23, wherein
a) the probe comprises a stem loop structure and one or more spacer sequences is located between the first target binding site and the stem-loop structure, within the stem-loop structure, 5′ of the stem loop structure, in the stem and/or in the loop; or b) the probe is a partially double-stranded structure and comprises one or more spacer sequences in the first strand between the first target binding site and the double-stranded region or 5′ to the double-stranded region, or in the second strand 5′ to the double-stranded region or 3′ to the double-stranded region. 25. The method of claim 23 or 24, wherein the probe comprises a third binding site, and further comprises one or more spacer sequences 3′ or 5′ to said third binding site, preferably wherein said one or more spacer sequences are directly adjacent to said third binding site. 26. The method of claim 23, wherein
a) the probe comprises a stem loop structure and the one or more spacer sequences is located 3′ of the stem of the stem-loop structure; or b) the probe is a partially double-stranded structure and comprises one or more spacer sequences in the first strand 3′ of the double-stranded region. 27. The method of any one of claims 23 to 26, wherein the spacer sequence is a capture sequence. 28. The method of claim 27, wherein the capture sequence hybridises to a cognate complementary binding site provided on a solid surface. 29. The method of claim 27, wherein the capture sequence or a complementary oligonucleotide hybridised thereto is attached to an affinity binding moiety. 30. The method of any one of claims 27 to 29, wherein
a) the probe comprises a stem loop structure and a capture sequence is provided 5′ of the stem of the stem-loop structure; or
b) the probe is a partially double-stranded structure and comprises a capture sequence in the second strand 5′ of the double-stranded region. 31. The method of any one of claims 23 to 30, wherein the spacer sequence is or comprises a tag sequence or a complement thereof, wherein the tag sequence is selected from a detection sequence or an identification sequence element or a binding site for a primer or detection probe. 32. The method of claim 31, wherein the tag sequence is an identification element for the target ROI or a sample identification sequence. 33. The method of any one of claims 23 to 26, wherein the probe comprises a stem loop structure and the spacer sequence is located within the loop of the stem-loop structure and is located 5′ of the second binding site thereby creating a gap between the respective ends of the extended strand when they are both hybridised to the second binding site in step (e), and wherein the gap is filled prior to ligation by one or more gap oligonucleotides which hybridise in the gap between the ends of the extended strand, or by gap-fill extension of the 3′ end of the hybridised extended strand using a polymerase, such that the extended strand and any gap oligonucleotides if present may be ligated into a circular molecule comprising the complement of the target fragment. 34. The method of claim 33, wherein the gap oligonucleotide(s) comprise(s) a tag sequence complementary to a tag sequence complement in the spacer sequence. 35. The method of claim 33, wherein the gap oligonucleotide comprises a region which is not complementary to the spacer sequence, and wherein the region of non-complementarity comprises a tag sequence. 36. The method of any one of claims 33 to 35, wherein the gap oligonucleotide comprises a detection sequence or an identification sequence element, or a binding site for an amplification primer or a detection probe. 37. The method of claim 36, wherein the amplification primer is a universal amplification primer. 38. The method of any one of claims 33 to 37, which is performed in multiplex using a plurality of different probes and wherein the gap oligonucleotide for each probe comprises the same tag sequence, preferably the same primer binding site. 39. The method of any one of claims 33 to 38, wherein the gap oligonucleotide is pre-hybridised to the probe prior to contacting the probe with the target nucleic acid molecule. 40. The method of any one of claims 33 to 38, wherein the gap oligonucleotide is separately provided at the same time or after contacting the probe with the target molecule. 41. The method of any one of claims 1 to 40, wherein the probe comprises a stem loop structure and the loop of the stem-loop structure comprises a region of intramolecular complementarity such that it is able to form a duplex within the loop, preferably wherein the duplex within the loop comprises a cleavage site. 42. The method of any one of claims 1 to 9 or 11 to 41, wherein cleavage is enzymatic cleavage. 43. The method of any one of claims 1 to 9 or 11 to 42, wherein the cleavage site is recognised by one or more enzymes capable of cleaving nucleic acid molecules. 44. The method of claim 43, wherein said one or more enzymes is a nickase or a restriction endonuclease. 45. The method of claim 44, wherein the nickase enzyme is removed from the assay or inactivated following unfolding of the probe. 46. The method of claim 42, wherein the enzymatic cleavage comprises a uracil-DNA glycosylase (UNG) enzyme in combination with an endonuclease enzyme capable of recognising apurinic/apyrimidinic sites of dsDNA. 47. The method of claim 46, wherein the endonuclease enzyme is capable of recognising apurinic/apyrimidinic sites of dsDNA is endonuclease IV. 48. The method of any one of claims 1 to 47, wherein amplification of the circularised extended strand of the probe is by PCR or Rolling Circle Amplification (RCA). 49. The method of claim 48, wherein amplification is by RCA, and wherein amplification further comprises a second round of RCA. 50. The method of any one of claims 1 to 49 further comprising a step of detecting and/or analysing the circularised extended strand or an amplicon thereof. 51. The method of claim 50, wherein the amplified product is detected by hybridising a detection probe labelled with a directly or indirectly detectable label to the amplification product. 52. The method of claim 51, wherein the detection label is a fluorescent label. 53. The method of claim 51, wherein the extended strand or amplicon thereof is detected or analysed by sequencing. 54. A probe for use in the method of any one of claims 1 to 8, or 11 to 53, said probe comprising a stem-loop structure and a single-stranded region at the 3′ end, wherein said 3′ end region comprises a first target-binding site which is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a flanking sequence flanking the 3′ end of the ROI, and said loop comprises a second binding site which is homologous to a flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence, and wherein said stem-loop structure further comprises a cleavage site 3′ of said second binding site such that cleavage allows the loop to open. 55. A probe for use in the method of any one of claims 1 or 9 to 53, said probe being a partially double-stranded construct having a first strand comprising single-stranded 3′ end region comprising a first target-binding site at the 3′ end thereof, wherein said first target binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a flanking sequence flanking the 3′ end of the ROI, and a second strand hybridised to said first strand at the 5′ end thereof and comprising a single-stranded 3′ end region comprising a second binding site, wherein the second binding site is homologous to a flanking sequence flanking the 5′ end of the ROI and is capable of hybridising to a complement of said 5′ flanking sequence. 56. A probe for use in the method of any one of claims 14 to 53, said probe comprising
(i) a first target-binding site at a 3′ end region of said probe, which binding site is capable of hybridising to a complementary binding site in the target molecule, said complementary binding site being a 3′ flanking sequence flanking the 3′ end of the ROI, and is capable of being extended to generate a complement of the target molecule in a target-templated extension reaction, said target complement comprising the complement of said 3′ flanking sequence and the ROI; and
(ii) a second binding site which is homologous to the ROI or to the 5′ end thereof and is capable of hybridising to a complement of the ROI or the 5′ end thereof;
(iii) a further probe sequence, lying 3′ of the second binding site, which is homologous to the 3′ flanking sequence and complementary to the first binding site;
wherein said probe is provided as an oligonucleotide comprising a stem-loop structure which comprises the second binding site and the further probe sequence in the loop of the structure and further comprises a cleavage site 3′ of the second binding site and further probe sequence, which is cleavable to open the loop to render the second binding site and further probe sequence available for binding, and a single-stranded region at the 3′ end of the probe comprising the first binding site at the 3′ end thereof; or
wherein said probe is provided as a partially double-stranded construct comprising a first strand comprising a single-stranded 3′ end region comprising the first binding site at the 3′ end thereof and a second strand hybridised at the 5′ end of said first strand and comprising a single-stranded 3′ end region comprising the second binding site and the further probe sequence at the 3′ end of the single-stranded region. 57. The probe of claim 54, wherein the probe is as defined in any one of claim 4 to 7, 15 to 18, 23 to 33 or 41. 58. The probe of claim 55, wherein the probe is as defined in any one of claims 11, 13, 15 or 23 to 32. 59. The probe of claim 56, wherein the probe is as defined in any one of claim 16 to 18, 23 to 33 or 41. 60. A kit for selecting a target region of interest in a target nucleic acid molecule, said kit comprising:
(a) a probe as defined in any one of claims 54 to 59; and optionally one or more further components selected from: (b) means for cleaving the cleavage site within the stem-loop structure of the probe; (c) means for cleaving the second cleavage site; (d) means for degrading the 3′ end of the extended probe, e.g. when the 5′ flanking sequence is internal to the 5′ end of the target molecule; (e) means for extending the probe, e.g. a polymerase enzyme; (f) a ligase enzyme; (g) one or more gap oligonucleotides; (h) means for amplification of the circularised extended strand;
(i) means for detecting the circularised extended strand or an amplicon thereof. | 1,600 |
1,052 | 16,064,302 | 1,612 | The present invention relates to a microstructure including a biocompatible polymer or an adhesive and to a method for manufacturing the same. The present inventors optimized the aspect ratio according to the type of each microstructure, thereby ensuring the optimal tip angle and the diameter range for skin penetration. Especially, the B-type to D-type microstructures of the present invention minimize the penetration resistance due to skin elasticity at the time of skin attachment, thereby increasing the penetration rate of the structures (60% or higher) and the absorption rate of useful ingredients into the skin. In addition, the D-type microstructure of the present invention maximizes the mechanical strength of the structure by applying a triple structure, and thus can easily penetrate the skin. When the plurality of microstructures are arranged in a hexagonal arrangement type, a uniform pressure can be transmitted to the whole microstructures on the skin. | 1. A microstructure comprising a biocompatible polymer or an adhesive, wherein the aspect ratio (w:h), configured of the diameter (w) of the bottom surface of the microstructure and the height (h) of the microstructure, is 1:5 to 1:1.5, and the angle of a distal tip is 10° to 40°. 2. The microstructure of claim 1, wherein the biocompatible polymer is at least one polymer selected from the group consisting of hyaluronic acid (HA), carboxymethyl cellulose (CMC), alginic acid, pectin, carrageenan, chondroitin (sulfate), dextran (sulfate), chitosan, polylysine, collagen, gelatin, carboxymethyl chitin, fibrin, agarose, pullulan polylactide, polyglycolide (PGA), polylactide-glycolide copolymer (PLGA), pullulan polyanhydride, polyorthoester, polyetherester, polycaprolactone, polyesteramide, poly(butyric acid), poly(valeric acid), polyurethane, polyacrylate, ethylene-vinyl acetate polymer, acrylic substituted cellulose acetate, non-degradable polyurethane, polystyrene, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonate polyolefin, polyethylene oxide, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polymethacrylate, hydroxypropyl methylcellulose (HPMC), ethylcellulose (EC), hydroxypropyl cellulose (HPC), cyclodextrin, copolymers of monomers forming these polymers, and cellulose. 3. The microstructure of claim 1, wherein the adhesive is at least one material selected from the group consisting of silicone, polyurethane, hyaluronic acid, a physical adhesive (Gecko), a polyacrylic material, ethylcellulose, hydroxymethyl cellulose, ethylene vinyl acetate, and polyisobutylene. 4. The microstructure of claim 1, wherein the aspect ratio of the microstructure is 1:5 to 1:2. 5. The microstructure of claim 1, wherein the height of the microstructure is 80 to 1500 μm. 6. The microstructure of claim 1, wherein the diameter (t) of the distal tip is 2 to 20 μm. 7. The microstructure of claim 1, wherein the microstructure has any one of A-type to D-type shapes in FIGS. 1a to 1 d. 8. The microstructure of claim 1, wherein the microstructure further comprises a metal, a polymer, or an adhesive. 9. The microstructure of claim 1, wherein the microstructure further comprises a useful ingredient other than the biodegradable polymer and the adhesive. 10. A method for manufacturing a microstructure, the method comprising:
(a) supplying a biodegradable polymer or an adhesive into a micro-mold; (b) injecting the biodegradable polymer or adhesive into a hole of the micro-mold; (c) drying the biodegradable polymer or adhesive; and (d) separating the dried biocompatible polymer or adhesive from the micro-mold to form a microstructure. 11. The method of claim 10, wherein step (c) is carried out (i) at room temperature for 36 to 60 hours, (ii) at 40 to 60° C. for 5 to 16 hours, or (iii) at 60 to 80° C. for 2 to 4 hours. 12. The method of claim 10, wherein, after the biodegradable polymer is supplied into the micro-mold, the injection is carried out by (i) applying a centrifugal force of 800 to 1000 g to the micro-mold or (ii) under a pressure of 500 to 860 mmHg. 13. The method of claim 10, wherein the biocompatible polymer is at least one polymer selected from the group consisting of hyaluronic acid (HA), carboxymethyl cellulose (CMC), alginic acid, pectin, carrageenan, chondroitin (sulfate), dextran (sulfate), chitosan, polylysine, collagen, gelatin, carboxymethyl chitin, fibrin, agarose, pullulan polylactide, polyglycolide (PGA), polylactide-glycolide copolymer (PLGA), pullulan polyanhydride, polyorthoester, polyetherester, polycaprolactones, polyesteramide, poly(butyric acid), poly(valeric acid), polyurethane, polyacrylate, ethylene-vinyl acetate polymer, acrylic substituted cellulose acetate, non-degradable polyurethane, polystyrene, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonate polyolefin, polyethylene oxide, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polymethacrylate, hydroxypropyl methylcellulose (HPMC), ethylcellulose (EC), hydroxypropyl cellulose (HPC), cyclodextrin, copolymers of monomers forming these polymers, and cellulose. 14. The method of claim 13, wherein the hyaluronic acid has a molecular weight of 240 to 490 kDa. 15. The method of claim 10, wherein, in step (a), the solid content of the biodegradable polymer is 1 to 30% (w/v) on the basis of the entire composition of the microstructure. 16. The method of claim 10, wherein the adhesive is at least one material selected from the group consisting of silicone, polyurethane, hyaluronic acid, a physical adhesive (Gecko), a polyacrylic material, ethylcellulose, hydroxymethyl cellulose, ethylene vinyl acetate, and polyisobutylene. 17. The method of claim 10, wherein a plurality of microstructures are arranged in a square or hexagonal shape. 18. The method of claim 17, wherein the plurality of microstructures are arranged at intervals (p) of 250 to 1500 μm. 19. A microstructure having any one of A-type to D-type shapes in FIGS. 1a to 1 d. | The present invention relates to a microstructure including a biocompatible polymer or an adhesive and to a method for manufacturing the same. The present inventors optimized the aspect ratio according to the type of each microstructure, thereby ensuring the optimal tip angle and the diameter range for skin penetration. Especially, the B-type to D-type microstructures of the present invention minimize the penetration resistance due to skin elasticity at the time of skin attachment, thereby increasing the penetration rate of the structures (60% or higher) and the absorption rate of useful ingredients into the skin. In addition, the D-type microstructure of the present invention maximizes the mechanical strength of the structure by applying a triple structure, and thus can easily penetrate the skin. When the plurality of microstructures are arranged in a hexagonal arrangement type, a uniform pressure can be transmitted to the whole microstructures on the skin.1. A microstructure comprising a biocompatible polymer or an adhesive, wherein the aspect ratio (w:h), configured of the diameter (w) of the bottom surface of the microstructure and the height (h) of the microstructure, is 1:5 to 1:1.5, and the angle of a distal tip is 10° to 40°. 2. The microstructure of claim 1, wherein the biocompatible polymer is at least one polymer selected from the group consisting of hyaluronic acid (HA), carboxymethyl cellulose (CMC), alginic acid, pectin, carrageenan, chondroitin (sulfate), dextran (sulfate), chitosan, polylysine, collagen, gelatin, carboxymethyl chitin, fibrin, agarose, pullulan polylactide, polyglycolide (PGA), polylactide-glycolide copolymer (PLGA), pullulan polyanhydride, polyorthoester, polyetherester, polycaprolactone, polyesteramide, poly(butyric acid), poly(valeric acid), polyurethane, polyacrylate, ethylene-vinyl acetate polymer, acrylic substituted cellulose acetate, non-degradable polyurethane, polystyrene, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonate polyolefin, polyethylene oxide, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polymethacrylate, hydroxypropyl methylcellulose (HPMC), ethylcellulose (EC), hydroxypropyl cellulose (HPC), cyclodextrin, copolymers of monomers forming these polymers, and cellulose. 3. The microstructure of claim 1, wherein the adhesive is at least one material selected from the group consisting of silicone, polyurethane, hyaluronic acid, a physical adhesive (Gecko), a polyacrylic material, ethylcellulose, hydroxymethyl cellulose, ethylene vinyl acetate, and polyisobutylene. 4. The microstructure of claim 1, wherein the aspect ratio of the microstructure is 1:5 to 1:2. 5. The microstructure of claim 1, wherein the height of the microstructure is 80 to 1500 μm. 6. The microstructure of claim 1, wherein the diameter (t) of the distal tip is 2 to 20 μm. 7. The microstructure of claim 1, wherein the microstructure has any one of A-type to D-type shapes in FIGS. 1a to 1 d. 8. The microstructure of claim 1, wherein the microstructure further comprises a metal, a polymer, or an adhesive. 9. The microstructure of claim 1, wherein the microstructure further comprises a useful ingredient other than the biodegradable polymer and the adhesive. 10. A method for manufacturing a microstructure, the method comprising:
(a) supplying a biodegradable polymer or an adhesive into a micro-mold; (b) injecting the biodegradable polymer or adhesive into a hole of the micro-mold; (c) drying the biodegradable polymer or adhesive; and (d) separating the dried biocompatible polymer or adhesive from the micro-mold to form a microstructure. 11. The method of claim 10, wherein step (c) is carried out (i) at room temperature for 36 to 60 hours, (ii) at 40 to 60° C. for 5 to 16 hours, or (iii) at 60 to 80° C. for 2 to 4 hours. 12. The method of claim 10, wherein, after the biodegradable polymer is supplied into the micro-mold, the injection is carried out by (i) applying a centrifugal force of 800 to 1000 g to the micro-mold or (ii) under a pressure of 500 to 860 mmHg. 13. The method of claim 10, wherein the biocompatible polymer is at least one polymer selected from the group consisting of hyaluronic acid (HA), carboxymethyl cellulose (CMC), alginic acid, pectin, carrageenan, chondroitin (sulfate), dextran (sulfate), chitosan, polylysine, collagen, gelatin, carboxymethyl chitin, fibrin, agarose, pullulan polylactide, polyglycolide (PGA), polylactide-glycolide copolymer (PLGA), pullulan polyanhydride, polyorthoester, polyetherester, polycaprolactones, polyesteramide, poly(butyric acid), poly(valeric acid), polyurethane, polyacrylate, ethylene-vinyl acetate polymer, acrylic substituted cellulose acetate, non-degradable polyurethane, polystyrene, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonate polyolefin, polyethylene oxide, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polymethacrylate, hydroxypropyl methylcellulose (HPMC), ethylcellulose (EC), hydroxypropyl cellulose (HPC), cyclodextrin, copolymers of monomers forming these polymers, and cellulose. 14. The method of claim 13, wherein the hyaluronic acid has a molecular weight of 240 to 490 kDa. 15. The method of claim 10, wherein, in step (a), the solid content of the biodegradable polymer is 1 to 30% (w/v) on the basis of the entire composition of the microstructure. 16. The method of claim 10, wherein the adhesive is at least one material selected from the group consisting of silicone, polyurethane, hyaluronic acid, a physical adhesive (Gecko), a polyacrylic material, ethylcellulose, hydroxymethyl cellulose, ethylene vinyl acetate, and polyisobutylene. 17. The method of claim 10, wherein a plurality of microstructures are arranged in a square or hexagonal shape. 18. The method of claim 17, wherein the plurality of microstructures are arranged at intervals (p) of 250 to 1500 μm. 19. A microstructure having any one of A-type to D-type shapes in FIGS. 1a to 1 d. | 1,600 |
1,053 | 14,651,887 | 1,657 | The present invention describes a composition comprising a mixture of human foetal keratinocyte cells and human foetal fibroblast cells, the ratio between said keratinocyte and fibroblast cells ranging from 0.75 to 2.5, preferably being 1:1 or 7:3. This composition is advantageously included in a bandage, said bandage preferably being sterile and packaged in a container impermeable to microorganisms. The present invention finally concerns the use of this composition as a drug, in particular for treating a skin defect (wound, burn or ulcer). | 1. A composition comprising a mixture of foetal human keratinocyte cells and foetal human fibroblastic cells, the ratio between said keratinocyte and fibroblast cells ranging from 0.75 to 2.5. 2. The composition according to claim 1, wherein said ratio is 1:1 or 7:3. 3. The composition according to claim 1 or 2, wherein said mixture of cells is integrated in a non-immunogenic extracellular matrix. 4. The composition according to claim 3, characterized in that it contains between 0.1 and 5.106 cells per mL or cm3 of matrix. 5. The composition according to claim 3 or 4, characterized in that said matrix contains at least one compound selected from the group consisting of: collagen, fibronectin, fibrin, and elastin, and mixtures thereof. 6. The composition according to claim 3 or 4, wherein said matrix is composed of human fibrinogen, human factor XIII, human fibronectin, plasminogen, bovine aprotinin and human thrombin. 7. A bandage containing the composition as defined in any one of claims 1 to 6. 8. The bandage according to claim 7, characterized in that it is sterile and packaged in a container impermeable to microorganisms. 9. A composition as defined and according to any one of claims 1 to 6, for use as a drug. 10. The composition as defined according to any one of claims 1 to 6, for use in the treatment of a skin defect, for example a wound, a burn or an ulcer. 11. A process for manufacturing a composition as defined in claims 1 to 6, comprising the step of mixing foetal human keratinocyte cells and foetal human fibroblastic cells with a non-immunogenic extracellular matrix, in a ratio ranging from 0.75 to 2.5. 12. The manufacturing process according to claim 11, characterized in that said ratio is 1:1 or 7:3. 13. The manufacturing process according to claim 11 or 12, characterized in that it comprises, prior to said mixing step, the following two steps:
a) obtaining foetal human keratinocyte cells, and
b) obtaining foetal human fibroblastic cells. 14. The manufacturing process according to claim 11 or 12, characterized in that it comprises, prior to the mixing step, the following two steps:
a) obtaining a bank of foetal human keratinocyte cells, and
b) obtaining a bank of foetal human fibroblastic cells. | The present invention describes a composition comprising a mixture of human foetal keratinocyte cells and human foetal fibroblast cells, the ratio between said keratinocyte and fibroblast cells ranging from 0.75 to 2.5, preferably being 1:1 or 7:3. This composition is advantageously included in a bandage, said bandage preferably being sterile and packaged in a container impermeable to microorganisms. The present invention finally concerns the use of this composition as a drug, in particular for treating a skin defect (wound, burn or ulcer).1. A composition comprising a mixture of foetal human keratinocyte cells and foetal human fibroblastic cells, the ratio between said keratinocyte and fibroblast cells ranging from 0.75 to 2.5. 2. The composition according to claim 1, wherein said ratio is 1:1 or 7:3. 3. The composition according to claim 1 or 2, wherein said mixture of cells is integrated in a non-immunogenic extracellular matrix. 4. The composition according to claim 3, characterized in that it contains between 0.1 and 5.106 cells per mL or cm3 of matrix. 5. The composition according to claim 3 or 4, characterized in that said matrix contains at least one compound selected from the group consisting of: collagen, fibronectin, fibrin, and elastin, and mixtures thereof. 6. The composition according to claim 3 or 4, wherein said matrix is composed of human fibrinogen, human factor XIII, human fibronectin, plasminogen, bovine aprotinin and human thrombin. 7. A bandage containing the composition as defined in any one of claims 1 to 6. 8. The bandage according to claim 7, characterized in that it is sterile and packaged in a container impermeable to microorganisms. 9. A composition as defined and according to any one of claims 1 to 6, for use as a drug. 10. The composition as defined according to any one of claims 1 to 6, for use in the treatment of a skin defect, for example a wound, a burn or an ulcer. 11. A process for manufacturing a composition as defined in claims 1 to 6, comprising the step of mixing foetal human keratinocyte cells and foetal human fibroblastic cells with a non-immunogenic extracellular matrix, in a ratio ranging from 0.75 to 2.5. 12. The manufacturing process according to claim 11, characterized in that said ratio is 1:1 or 7:3. 13. The manufacturing process according to claim 11 or 12, characterized in that it comprises, prior to said mixing step, the following two steps:
a) obtaining foetal human keratinocyte cells, and
b) obtaining foetal human fibroblastic cells. 14. The manufacturing process according to claim 11 or 12, characterized in that it comprises, prior to the mixing step, the following two steps:
a) obtaining a bank of foetal human keratinocyte cells, and
b) obtaining a bank of foetal human fibroblastic cells. | 1,600 |
1,054 | 15,541,822 | 1,618 | The invention relates generally to a formulation in which metal tungstate or metal molybdate particles are encapsulated within biocompatible, diseased cell-targeting polymeric coatings. Such formulations render metal tungstate or metal molybdate particles suitable for in vivo biomedical imaging and therapeutic applications. | 1. A formulation, comprising a crystalline particle or particle aggregate encapsulated within a biocompatible polymeric coating material, wherein the crystalline particle or particle aggregate is a metal tungstate material, a metal molybdate material, or a combination thereof. 2. The formulation of claim 1, wherein said metal tungstate material (Mx(WO4)y) comprises a metal compound (M) selected from the “Alkaline Earth Metal”, “Transition Metal” or “Poor Metal” group of elements in the periodic table, or an atomic mixture thereof. 3. The formulation of claim 1, wherein said metal tungstate material is calcium tungstate (CaWO4). 4. The formulation of claim 1, wherein said particle is a composite material comprising metal tungstate and other biocompatible organic or inorganic compound. 5. (canceled) 6. The formulation of claim 1, wherein said metal molybdate material (Mx(MoO4)y) comprises a metal compound (M) selected from the “Alkaline Earth Metal”, “Transition Metal” or “Poor Metal” group of elements in the periodic table, or an atomic mixture thereof. 7. The formulation of claim 1, wherein said particle is a composite material comprising metal molybdate and other biocompatible organic or inorganic compound. 8. The formulation of claim 1, wherein the mean largest dimension of said particle is in the range between about 1 nm and about 50,000 nm in its unaggregated state. 9. (canceled) 10. The formulation of claim 1, wherein the mean diameter of said particle or particle aggregate is in the range between about 1 nm and about 500 nm. 11. The formulation of claim 1, wherein said coating material encapsulated with a biocompatible amphiphilic block copolymer comprising one or more of the following polymer components: polyethylene glycol (PEG), poly(-lactic acid) (PLA), poly(lactic acid-co-glycolic acid) (PLGA), poly(caprolactone) (PCL), poly(styrene) (PS), poly(n-butyl acrylate) (PnBA). 12. The formulation of claim 1, wherein said particle or said coating material is functionalized with a targeting agent specific to diseased cells in humans or animals. 13. The formulation of claim 1, wherein said particle or coating material is functionalized with folic acid. 14. The formulation of claim 12, wherein said diseased cells are cancer cells. 15. A polymer-encapsulated radio-luminescent particle, wherein said particle is a calcium tungstate (CaWO4) particle. 16. The polymer-encapsulated radio-luminescent particle of claim 15, wherein the mean diameter of said particle material is in the range between about 0.1 and 1000 nm. 17. The polymer-encapsulated radio-luminescent particle of claim 15, wherein the mean diameter of said particle is ranged between about 1 and 10 μm. 18. The polymer-encapsulated radio-luminescent particle of claim 15, wherein the mean diameter of said particle is in the range between about 1 and 200 nm. 19. The polymer-encapsulated radio-luminescent particle of claim 15, wherein said polymer-encapsulated particle is functionalized or conjugated with a targeting agent specific to a cancer. 20. The polymer-encapsulated radio-luminescent particle of claim 19, wherein said targeting agent is a biological molecule having specific affinity to said cancer so as to enhance the delivery of said polymer-encapsulated particles to tumor cells. 21. The polymer-encapsulated radio-luminescent particle of claim 15, wherein said polymer-encapsulated particle is functionalized with folic acid. 22. The polymer-encapsulated radio-luminescent particle of claim 15, wherein said particle is functionalized with polyethylene glycol (PEG), poly(lactic acid-co-glycolic acid) (PLGA), or a combination thereof. | The invention relates generally to a formulation in which metal tungstate or metal molybdate particles are encapsulated within biocompatible, diseased cell-targeting polymeric coatings. Such formulations render metal tungstate or metal molybdate particles suitable for in vivo biomedical imaging and therapeutic applications.1. A formulation, comprising a crystalline particle or particle aggregate encapsulated within a biocompatible polymeric coating material, wherein the crystalline particle or particle aggregate is a metal tungstate material, a metal molybdate material, or a combination thereof. 2. The formulation of claim 1, wherein said metal tungstate material (Mx(WO4)y) comprises a metal compound (M) selected from the “Alkaline Earth Metal”, “Transition Metal” or “Poor Metal” group of elements in the periodic table, or an atomic mixture thereof. 3. The formulation of claim 1, wherein said metal tungstate material is calcium tungstate (CaWO4). 4. The formulation of claim 1, wherein said particle is a composite material comprising metal tungstate and other biocompatible organic or inorganic compound. 5. (canceled) 6. The formulation of claim 1, wherein said metal molybdate material (Mx(MoO4)y) comprises a metal compound (M) selected from the “Alkaline Earth Metal”, “Transition Metal” or “Poor Metal” group of elements in the periodic table, or an atomic mixture thereof. 7. The formulation of claim 1, wherein said particle is a composite material comprising metal molybdate and other biocompatible organic or inorganic compound. 8. The formulation of claim 1, wherein the mean largest dimension of said particle is in the range between about 1 nm and about 50,000 nm in its unaggregated state. 9. (canceled) 10. The formulation of claim 1, wherein the mean diameter of said particle or particle aggregate is in the range between about 1 nm and about 500 nm. 11. The formulation of claim 1, wherein said coating material encapsulated with a biocompatible amphiphilic block copolymer comprising one or more of the following polymer components: polyethylene glycol (PEG), poly(-lactic acid) (PLA), poly(lactic acid-co-glycolic acid) (PLGA), poly(caprolactone) (PCL), poly(styrene) (PS), poly(n-butyl acrylate) (PnBA). 12. The formulation of claim 1, wherein said particle or said coating material is functionalized with a targeting agent specific to diseased cells in humans or animals. 13. The formulation of claim 1, wherein said particle or coating material is functionalized with folic acid. 14. The formulation of claim 12, wherein said diseased cells are cancer cells. 15. A polymer-encapsulated radio-luminescent particle, wherein said particle is a calcium tungstate (CaWO4) particle. 16. The polymer-encapsulated radio-luminescent particle of claim 15, wherein the mean diameter of said particle material is in the range between about 0.1 and 1000 nm. 17. The polymer-encapsulated radio-luminescent particle of claim 15, wherein the mean diameter of said particle is ranged between about 1 and 10 μm. 18. The polymer-encapsulated radio-luminescent particle of claim 15, wherein the mean diameter of said particle is in the range between about 1 and 200 nm. 19. The polymer-encapsulated radio-luminescent particle of claim 15, wherein said polymer-encapsulated particle is functionalized or conjugated with a targeting agent specific to a cancer. 20. The polymer-encapsulated radio-luminescent particle of claim 19, wherein said targeting agent is a biological molecule having specific affinity to said cancer so as to enhance the delivery of said polymer-encapsulated particles to tumor cells. 21. The polymer-encapsulated radio-luminescent particle of claim 15, wherein said polymer-encapsulated particle is functionalized with folic acid. 22. The polymer-encapsulated radio-luminescent particle of claim 15, wherein said particle is functionalized with polyethylene glycol (PEG), poly(lactic acid-co-glycolic acid) (PLGA), or a combination thereof. | 1,600 |
1,055 | 14,240,825 | 1,616 | The present invention relates to a composition comprising a plant growth regulator and/or fungicide and/or insecticide and/or acaricide and the preparation process thereof. Additionally, the present invention discloses the use of a plant growth regulator and/or fungicide and/or insecticide and/or acaricide to prepare a composition and the use of a composition to ensure high yield per area of a corn crop. Furthermore, the present invention provides methods to reduce excessive growth of corn plants, to strengthen stems, the root system and adventitious roots of corn plants, to combat diseases and pests and to confer resistance to pest control, as well as a method to identify a composition that ensures high corn crop yield. | 1. A composition, comprising:
a plant growth regulator and a fungicide; or a plant growth regulator and an insecticide; or a plant growth regulator and an acaricide; or a plant growth regulator and a fungicide and an insecticide; or a plant growth regulator and an insecticide and an acaricide; or a plant growth regulator and a fungicide and an acaricide. 2. The composition according to claim 1, wherein the plant growth regulator is selected from the group consisting of ethephon, mepiquat chloride, Ca-prohexadione, ancymidol, flurprimidol, chlormequat, dikegulac sodium, mefluidide, uniconazole, paclobutazole, 1-naphthaleneacetic acid (NAA), 1-naphthalene acetamide, carbaryl, 6-benzyladenine (6BA), etichlozate, sulphocarbamide, ethylene, gibberellic acid (GA3), GA47, choline chloride, benzyladenine, cyanamide, cynetine and acybenzolar-S-methyl. 3. The composition according to claim 2, wherein the plant growth regulator is ethephon. 4. The composition according to claim 1, wherein the fungicide is selected from the group consisting of triazoles, strobilurins, benzimidazoles, acylanilides, carboxanilides, benzamides and pyrazoles. 5. The composition according to claim 3, wherein the fungicide is selected from the group consisting of triazoles, strobilurins and carboxanilides. 6. The composition according to claim 4, characterized in that:
the triazoles are selected from the group consisting of: azaconazole, bitertanol, bromuconazole, cyproconazole, difenoconazole, diniconazole, epoxyconazole, etaconazole, fenbuconazole, fluquinconazole, flusilazole, flutriafole, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propiconazole, prothioconazole, simeconazole, tebuconazole, tetraconazole, triadimefon, triadimenol, and triticonazole; the strobilurins are selected from the group consisting of: azoxystrobin, enestrobin, picoxystrobin, piraoxystrobin, pyraclostrobin, pyrametostrobin, kresoxim-methyl, trifloxystrobin, dimoxystrobin, metominostrobin, orysastrobin, famoxadone, fluoxastrobin, fenamidone, and pyribencarb; the benzimidazoles are selected from the group consisting of: benomyl, carbendazim, fuberidazole, thiabendazole, thiophanate, and tiophanate-methyl; the acylanilides are selected from the group consisting of: benalaxyl, furalaxyl, metalaxyl, metalaxyl-M, and mefenoxam; the benzamides are selected from the group consisting of: benodanil, fluopyram, flutolanil, and mepronil; the carboxanilides are selected from the group consisting of: fenfuram, carboxin, oxycarboxin, thifluzamide, bixafen, furametpyr, isopyrazam, penflufen, penthiopyrad, sedaxane, and boscalid; and the pyrazoles are selected from the group consisting of: fipronil and ethiprole. 7. The composition according to claim 1, wherein the insecticide is selected from the group consisting of anthranilamide, phthalic acid diamide, pyrazole analog, avermectins, benzoylurea, biological agents, bis(thiocarbamate), ketoenol, chlorocyclodiene, spinosyns, pyridyloxypropyl ether, phenylpyrazole, phenylthiourea, carbamates, neonicotinoids, nicotinoids, organophosphorous, oxadiazine, pyrazole, and pyrethroid. 8. The composition according to claim 7, wherein:
the anthranilamide are selected from the group consisting of: chlorantraniliprole and cyantraniliprole; the phthalic acid diamide is flubendiamide; the pyrazole analog is chlorfenapyr; the avermectin is abamectin; the benzoylurea is selected from the group consisting of: chlorfluazuron, diflubenzuron, flufenoxuron, lufenuron, novaluron, eflubenzuron, and triflumuron; the biological agent is selected from the group consisting of: Bacillus thuringiensis, Baculovirus anticarsia, and Metarhizium anisopliae; the bis(thiocarbamate) is cartap hydrochloride; the ketoenol is selected from the group consisting of: spiromesifen, spirodiclofen, and spirotetramat; the chlorocyclodiene is endosulfan; the spynosyns is spinosad; the pyridyloxypropyl ether is pyriproxyfen; the phenylpyrazole is ethiprole; the phenylthiourea is diafentiuron; the carbamates are selected from the group consisting of: benfuracarb, carbofuran, carbosulfan, furathiocarb, alanycarb, aldicarb, methomyl, and thiodicarb; the neonicotinoid is selected from the group consisting of: acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, and clothianidin, the nicotinoid is flonicamid; the organophosphorous is selected from the group consisting of: acephate, cadusafos, chlorpyrifos, diazinon, dimethoate, disulfoton, ethion, fenitrothion, fenthion, fentoato, phorate, phosmet, fosthiazate, malathion, methamidophos, methidathion, mevinphos, parathion-methyl, pyridaphenthion, pirimiphos-methyl, profenophos, prothiophos, tebupyrimphos, terbuphos, triazophos, and trichlorphon; the oxadiazine is indoxacarb, the pyrazole is fipronil; and the pyrethroid is selected from the group consisting of: alpha-cypermethrin, betacyfluthrin, beta-cypermethrin, bifenthrin, cyfluthrin, cypermethrin, deltamethrin, esfenvalerate, fenpropatrina, fluvalinate, gamma-cyhalothrin, lambda-cyhalothrin, permethrin, and zeta-cypermethrin. 9. The composition according to claim 1, wherein the acaricide is selected from the group consisting of avermectins, bis(arylformamidine), ketoenol, chlorodiphenylsulfone, diacyl-hydrazine, diphenyl oxazoline, dinitrophenol, phenylthiourea, oxazolidinadione, pyrazole, pyridazinone, alkyl sulphite, tetrazine, thiadiazinone, and thiazolidine carboxamide. 10. The composition according to claim 9, wherein:
the avermectins is abamectin; the bis(arylformamidine) is amitraz; the ketoenol is selected from the group consisting of: spirodiclofen, spirotetramat, and spiromesifen; the chlorodiphenylsulfone is tetradifon; the diacyl-hydrazine is chromafenozide; the diphenyl oxazoline is etoxazole; the dinitrophenol is dinocap; the phenylthiourea is diafentiuron; the oxazolidinedione is famoxadone; the pyrazole is fenpyroximate; the pyridazinone is pyridaben; the alkyl sulphite is propargite; the tetrazine is clofentezine; the thiadiazinone is buprofezin; and the thiazolidine carboxamide is hexythiazox. 11. The composition according to claim 1, wherein the plant growth regulator is present in an amount ranging from about 1 to about 1000 g a.i./ha, more preferably, from about 2.5 to about 500 g a.i./ha. 12. The composition according to claim 1, wherein in that the fungicide, insecticide or acaricide is present in an amount ranging from about 10 to about 500 g/L or g/kg of formulated product, more preferably, from about 20 to about 400 g/L or g/kg of formulated product. 13. The composition according to claim 1, wherein it is applied to crop plants at a density equal to or greater than 100,000 plants per hectare. 14. The composition according to claim 13, wherein the plant is a corn plant. 15. The composition according to claim 14, wherein the corn plant is conventional corn, transgenic corn, mutagenic corn, modified corn, sweet corn, or popcorn. 16. Use of a
a plant growth regulator and a fungicide; or a plant growth regulator and an insecticide; or a plant growth regulator and an acaricide; or a plant growth regulator and a fungicide and an insecticide; or a plant growth regulator and an insecticide and an acaricide; or a plant growth regulator and a fungicide and an acaricide for a preparation of a composition. 17. Use of a composition of claim 1, for ensuring high yield per area of a corn crop, together with controlling diseases and parasites that affect corn crops. 18. The use according to claim 17, wherein the parasites are selected from the group consisting of caterpillars (Spodoptera frugiperda, Spodoptera latifacea, Spodoptera eridanea, Spodoptera sp.), leafworms (Mocis latipes), leafhoppers (Deois flavopicta), corn planthoppers (Peregrinus maidis and Dalbulus maidis), corn aphids (Rhopalosiphum maidis), sugarcane borer (Diatraea saccharalis), corn earworms (Helicoverpa zea), pentatomid bugs (Dichelops melacanthus and Dichelops furcatus), neotropical brown stinkbugs (Euschistus heros, Nezara viridula, Piezodorus guildine), leaffooted bugs (Leptoglossus zonatus), squash bug (Phthia pitta), armyworms (Pseudaletia sp.) and mites. 19. The use according to claim 17, wherein the diseases are selected from the group consisting of spot blotch (Exerohilum turcicum), white leaf spot or phaeospheria leaf spot (Phaeosphaeria maydis), diplodia spot (Diplodia macrospora), cercospora leaf spot (Cercospora zea-maydis), anthracnose (Colletotrichum graminicola), polissora rust (Puccinia polysora), tropical rust (Physopella zeae) and common rust (Puccinia sorghi). 20. The use according to claim 17, wherein in that said high yield per area of a corn crop relates to a density greater than 100.000 corn plants per hectare. 21. The use according to claim 17, wherein in that the plant is conventional corn, transgenic corn, mutagenic corn, modified corn, sweet corn, or popcorn. 22. A method for reducing excessive growth of corn plants, comprising the step of an application of an agronomically effective amount of a composition, as defined in claim 1, to the plant, its habitat, progeny or propagation material or combination thereof. 23. A method for strengthening the stem of corn plants, comprising the step of an application of an agronomically effective amount of a composition, as defined in claim 1, to the plant, its habitat, progeny or propagation material or combination thereof. 24. A method for strengthening the root system of corn plants, comprising the step of an application of an agronomically effective amount of a composition, as defined in claim 1, to the plant, its habitat, progeny or propagation material or combination thereof. 25. A method for strengthening the adventitious roots of corn plants, characterized by comprising the application of an agronomically effective amount of a composition, as defined in claim 1, to the plant, its habitat, progeny or propagation material or combination thereof. 26. A method for combating pests in plant crops, wherein agronomically effective amount of a composition, as defined in claim 1, is allowed to act to said pests or their habitat or combination thereof. 27. A method for conferring resistance to pest control in a corn crop, comprising an application of an agronomically effective amount of a composition, as defined in claim 1, to the corn crop. 28. The method according to claim 27, wherein the pests are selected from the group consisting of caterpillars (Spodoptera frugiperda, Spodoptera latifacea, Spodoptera eridanea, Spodoptera sp.), leafworms (Mocis latipes), leafhoppers (Deois flavopicta), corn planthoppers (Peregrinus maidis and Dalbulus maidis), corn aphids (Rhopalosiphum maidis), sugarcane borer (Diatraea saccharalis), corn earworms (Helicoverpa zea), pentatomid bugs (Dichelops melacanthus and Dichelops furcatus), neotropical brown stinkbugs (Euschistus heros, Nezara viridula, Piezodorus guildine), leaffooted bugs (Leptoglossus zonatus), squash bug (Phthia picta), armyworms (Pseudaletia sp.) and mites. 29. A method for identifying a composition that ensures high yield of corn crops, comprising the steps of:
a) applying an agronomically effective amount of a candidate composition to one or more plants; b) sprouting said one or more plants; c) comparing the growth rate of said plants with the growth rate of untreated plants; and d) identifying the candidate composition as a composition that ensures high yield in corn crops and that additionally ensures control of diseases caused by parasites that attack corn plants, wherein said candidate composition comprises a plant growth regulator and fungicide(s), a plant growth regulator and insecticide(s), a regulator and insecticide and fungicide, wherein said high yield is related to a reduction of the excessive growth of corn plants, strengthening of the stem of corn plants, strengthening of the corn plant root system. 30. The method according to claim 29, wherein the candidate composition is applied in an amount ranging from 1 to 2000 g a.i./ha, more preferably from 2.5 to 1000 g a.i./ha. 31. A process for preparing a composition, comprising combining agronomically effective amounts of a
a plant growth regulator and a fungicide; or a plant growth regulator and an insecticide; or a plant growth regulator and an acaricide; or a plant growth regulator and a fungicide and an insecticide; or a plant growth regulator and an insecticide and an acaricide; or a plant growth regulator and a fungicide and an acaricide, as defined in claim 1, and, later, properly mixing them with carriers, adjuvants and/or excipients thereof, such as mineral oil, vegetal oil, silicone adjuvants, non-silicone adjuvants, soybean oil methylate esther, nonylphenol ethoxylate, ethoxylate alkyl esther of phosphoric acid, mixture of paraffin hydrocarbons, cyclic paraffins and saturated and unsaturated aromatics. 32. The process according to claim 31, wherein the plant growth regulator is selected from the group consisting of ethephon, mepiquat chloride, Ca-prohexadione, ancymidol, flurprimidol, chlormequat, dikegulac sodium, mefluidide, uniconazole, paclobutazole, 1-naphthaleneacetic acid (NAA), 1-naphthalene acetamide, carbaryl, 6-benzyladenine (6BA), etichlozate, sulphocarbamide, ethylene, gibberellic acid (GA3), GA47, choline chloride, benzyladenine, cyanamide, cynetine and acybenzolar-S-methyl. 33. The process according to claim 31, wherein the plant growth regulator is ethephon. 34. The process according to claim 31, wherein the fungicide is selected from the group consisting of triazoles, strobilurins, benzimidazoles, acylanilides, carboxanilides, benzamides and pyrazoles. 35. The process according to claim 34, wherein:
the triazoles are selected from the group consisting of: azaconazole, bitertanol, bromuconazole, cyproconazole, difenoconazole, diniconazole, epoxyconazole, etaconazole, fenbuconazole, fluquinconazole, flusilazole, flutriafole, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propiconazole, prothioconazole, simeconazole, tebuconazole, tetraconazole, triadimefon, triadimenol, and triticonazole; the strobilurins are selected from the group consisting of: azoxystrobin, enestrobin, picoxystrobin, piraoxystrobin, pyraclostrobin, pyrametostrobin, kresoxim-methyl, trifloxystrobin, dimoxystrobin, metominostrobin, orysastrobin, famoxadone, fluoxastrobin, fenamidone, and pyribencarb; the benzimidazoles are selected from the group consisting of: benomyl, carbendazim, fuberidazole, thiabendazole, thiophanate, and tiophanato-methyl; the acylanilides are selected from the group consisting of: benalaxyl, furalaxyl, metalaxyl, metalaxyl-M, and mefenoxam; the benzamides are selected from the group consisting of: benodanil, fluopyram, flutolanil, and mepronil; the carboxanilides are selected from the group consisting of: fenfuram, carboxin, oxycarboxin, thifluzamide, bixafen, furametpyr, isopyrazam, penflufen, penthiopyrad, sedaxane, and boscalid; and the pyrazoles are selected from the group consisting of: fipronil and ethiprole. 36. The process according to claim 31, wherein:
the insecticide is selected from the group consisting of anthranilamide, phthalic acid diamide, pyrazole analog, avermectins, benzoylurea, biological agent, bis(thiocarbamate), ketoenol, chloro-cyclodiene, spinosyns, pyridyloxypropyl ether, phenylpyrazole, phenylthiourea, carbamates, neonicotinoids, nicotinoids, organophosphorous, oxadiazine, pyrazole, and pyrethroid. 37. The process according to claim 36, wherein:
the anthranilamide are selected from the group consisting of: chlorantraniliprole and cyantraniliprole; the phthalic acid diamide is flubendiamide; the pyrazole analog is chlorfenapyr; the avermectin is abamectin; the benzoylurea is selected from the group consisting of: chlorfluazuron, diflubenzuron, flufenoxuron, lufenuron, novaluron, eflubenzuron, and triflumuron; the biological agent is selected from the group consisting of: Bacillus thuringiensis, Baculovirus anticarsia, and Metarhizium anisopliae; the bis(thiocarbamate) is cartap hydrochloride; the ketoenol is selected from the group consisting of: spiromesifen, spirodiclofen, spirotetramat; the chlorocyclodiene is endosulfan; the spynosyn is spinosad; the pyridyloxypropyl ether is pyriproxyfen; the phenylpyrazole is ethiprole; the phenylthiourea is diafentiuron; the carbamates are selected from the group consisting of: benfuracarb, carbofuran, carbosulfan, furathiocarb, alanycarb, aldicarb, methomyl, and thiodicarb; the neonicotinoid is selected from the group consisting of: acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, and clothianidin, the nicotinoid is flonicamid; the organophosphorous is selected from the group consisting of: acephate, cadusafos, chlorpyrifos, diazinon, dimethoate, disulfoton, ethion, fenitrothion, fenthion, fentoato, phorate, phosmet, fosthiazate, malathion, methamidophos, methidathion, mevinphos, parathion-methyl, pyridaphenthion, pirimiphos-methyl, profenophos, prothiophos, tebupyrimphos, terbuphos, triazophos, and trichlorphon; the oxadiazine is indoxacarb, the pyrazole is fipronil; and the pyrethroid is selected from the group consisting of: alpha-cypermethrin, betacyfluthrin, beta-cypermethrin, bifenthrin, cyfluthrin, cypermethrin, deltamethrin, esfenvalerate, fenpropatrina, fluvalinate, gamma-cyhalothrin, lambda-cyhalothrin, permethrin, and zeta-cypermethrin. 38. The process according to claim 31, wherein the acaricide is selected from the group consisting of avermectins, bis(arylformamidine), ketoenol, chlorodiphenylsulfone, diacyl-hydrazine, diphenyl oxazoline, dinitrophenol, phenylthiourea, oxazolidinadione, pyrazole, pyridazinone, alkyl sulphite, tetrazine, thiadiazinone, and thiazolidine carboxamide. 39. The process according to claim 38, wherein:
the avermectin is abamectin; the bis(arylformamidine) is amitraz; the ketoenol is selected from the group consisting of: spirodiclofen, spirotetramat, and spiromesifen; the chlorodiphenylsulfone is tetradifon; the diacyl-hydrazine is chromafenozide; the diphenyl oxazoline is etoxazole; the dinitrophenol is dinocap; the phenylthiourea is diafentiuron; the oxazolidinedione is famoxadone; the pyrazole is fenpyroximate; the pyridazinone is pyridaben; the alkyl sulphite is propargite; the tetrazine is clofentezine; the thiadiazinone is buprofezin; and the thiazolidine carboxamide is hexythiazox. 40. The process according to claim 31, wherein the plant growth regulator is present in an amount ranging from about 1 to about 1000 g a.i./ha, more preferably, about 2.5 to about 500 g a.i./ha. 41. The process according to claim 31, wherein the fungicide, insecticide or acaricide is present in an amount ranging from about 10 to about 500 g/L or g/kg of formulated product, more preferably, from about 20 to about 400 g/L or g/kg of formulated product. | The present invention relates to a composition comprising a plant growth regulator and/or fungicide and/or insecticide and/or acaricide and the preparation process thereof. Additionally, the present invention discloses the use of a plant growth regulator and/or fungicide and/or insecticide and/or acaricide to prepare a composition and the use of a composition to ensure high yield per area of a corn crop. Furthermore, the present invention provides methods to reduce excessive growth of corn plants, to strengthen stems, the root system and adventitious roots of corn plants, to combat diseases and pests and to confer resistance to pest control, as well as a method to identify a composition that ensures high corn crop yield.1. A composition, comprising:
a plant growth regulator and a fungicide; or a plant growth regulator and an insecticide; or a plant growth regulator and an acaricide; or a plant growth regulator and a fungicide and an insecticide; or a plant growth regulator and an insecticide and an acaricide; or a plant growth regulator and a fungicide and an acaricide. 2. The composition according to claim 1, wherein the plant growth regulator is selected from the group consisting of ethephon, mepiquat chloride, Ca-prohexadione, ancymidol, flurprimidol, chlormequat, dikegulac sodium, mefluidide, uniconazole, paclobutazole, 1-naphthaleneacetic acid (NAA), 1-naphthalene acetamide, carbaryl, 6-benzyladenine (6BA), etichlozate, sulphocarbamide, ethylene, gibberellic acid (GA3), GA47, choline chloride, benzyladenine, cyanamide, cynetine and acybenzolar-S-methyl. 3. The composition according to claim 2, wherein the plant growth regulator is ethephon. 4. The composition according to claim 1, wherein the fungicide is selected from the group consisting of triazoles, strobilurins, benzimidazoles, acylanilides, carboxanilides, benzamides and pyrazoles. 5. The composition according to claim 3, wherein the fungicide is selected from the group consisting of triazoles, strobilurins and carboxanilides. 6. The composition according to claim 4, characterized in that:
the triazoles are selected from the group consisting of: azaconazole, bitertanol, bromuconazole, cyproconazole, difenoconazole, diniconazole, epoxyconazole, etaconazole, fenbuconazole, fluquinconazole, flusilazole, flutriafole, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propiconazole, prothioconazole, simeconazole, tebuconazole, tetraconazole, triadimefon, triadimenol, and triticonazole; the strobilurins are selected from the group consisting of: azoxystrobin, enestrobin, picoxystrobin, piraoxystrobin, pyraclostrobin, pyrametostrobin, kresoxim-methyl, trifloxystrobin, dimoxystrobin, metominostrobin, orysastrobin, famoxadone, fluoxastrobin, fenamidone, and pyribencarb; the benzimidazoles are selected from the group consisting of: benomyl, carbendazim, fuberidazole, thiabendazole, thiophanate, and tiophanate-methyl; the acylanilides are selected from the group consisting of: benalaxyl, furalaxyl, metalaxyl, metalaxyl-M, and mefenoxam; the benzamides are selected from the group consisting of: benodanil, fluopyram, flutolanil, and mepronil; the carboxanilides are selected from the group consisting of: fenfuram, carboxin, oxycarboxin, thifluzamide, bixafen, furametpyr, isopyrazam, penflufen, penthiopyrad, sedaxane, and boscalid; and the pyrazoles are selected from the group consisting of: fipronil and ethiprole. 7. The composition according to claim 1, wherein the insecticide is selected from the group consisting of anthranilamide, phthalic acid diamide, pyrazole analog, avermectins, benzoylurea, biological agents, bis(thiocarbamate), ketoenol, chlorocyclodiene, spinosyns, pyridyloxypropyl ether, phenylpyrazole, phenylthiourea, carbamates, neonicotinoids, nicotinoids, organophosphorous, oxadiazine, pyrazole, and pyrethroid. 8. The composition according to claim 7, wherein:
the anthranilamide are selected from the group consisting of: chlorantraniliprole and cyantraniliprole; the phthalic acid diamide is flubendiamide; the pyrazole analog is chlorfenapyr; the avermectin is abamectin; the benzoylurea is selected from the group consisting of: chlorfluazuron, diflubenzuron, flufenoxuron, lufenuron, novaluron, eflubenzuron, and triflumuron; the biological agent is selected from the group consisting of: Bacillus thuringiensis, Baculovirus anticarsia, and Metarhizium anisopliae; the bis(thiocarbamate) is cartap hydrochloride; the ketoenol is selected from the group consisting of: spiromesifen, spirodiclofen, and spirotetramat; the chlorocyclodiene is endosulfan; the spynosyns is spinosad; the pyridyloxypropyl ether is pyriproxyfen; the phenylpyrazole is ethiprole; the phenylthiourea is diafentiuron; the carbamates are selected from the group consisting of: benfuracarb, carbofuran, carbosulfan, furathiocarb, alanycarb, aldicarb, methomyl, and thiodicarb; the neonicotinoid is selected from the group consisting of: acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, and clothianidin, the nicotinoid is flonicamid; the organophosphorous is selected from the group consisting of: acephate, cadusafos, chlorpyrifos, diazinon, dimethoate, disulfoton, ethion, fenitrothion, fenthion, fentoato, phorate, phosmet, fosthiazate, malathion, methamidophos, methidathion, mevinphos, parathion-methyl, pyridaphenthion, pirimiphos-methyl, profenophos, prothiophos, tebupyrimphos, terbuphos, triazophos, and trichlorphon; the oxadiazine is indoxacarb, the pyrazole is fipronil; and the pyrethroid is selected from the group consisting of: alpha-cypermethrin, betacyfluthrin, beta-cypermethrin, bifenthrin, cyfluthrin, cypermethrin, deltamethrin, esfenvalerate, fenpropatrina, fluvalinate, gamma-cyhalothrin, lambda-cyhalothrin, permethrin, and zeta-cypermethrin. 9. The composition according to claim 1, wherein the acaricide is selected from the group consisting of avermectins, bis(arylformamidine), ketoenol, chlorodiphenylsulfone, diacyl-hydrazine, diphenyl oxazoline, dinitrophenol, phenylthiourea, oxazolidinadione, pyrazole, pyridazinone, alkyl sulphite, tetrazine, thiadiazinone, and thiazolidine carboxamide. 10. The composition according to claim 9, wherein:
the avermectins is abamectin; the bis(arylformamidine) is amitraz; the ketoenol is selected from the group consisting of: spirodiclofen, spirotetramat, and spiromesifen; the chlorodiphenylsulfone is tetradifon; the diacyl-hydrazine is chromafenozide; the diphenyl oxazoline is etoxazole; the dinitrophenol is dinocap; the phenylthiourea is diafentiuron; the oxazolidinedione is famoxadone; the pyrazole is fenpyroximate; the pyridazinone is pyridaben; the alkyl sulphite is propargite; the tetrazine is clofentezine; the thiadiazinone is buprofezin; and the thiazolidine carboxamide is hexythiazox. 11. The composition according to claim 1, wherein the plant growth regulator is present in an amount ranging from about 1 to about 1000 g a.i./ha, more preferably, from about 2.5 to about 500 g a.i./ha. 12. The composition according to claim 1, wherein in that the fungicide, insecticide or acaricide is present in an amount ranging from about 10 to about 500 g/L or g/kg of formulated product, more preferably, from about 20 to about 400 g/L or g/kg of formulated product. 13. The composition according to claim 1, wherein it is applied to crop plants at a density equal to or greater than 100,000 plants per hectare. 14. The composition according to claim 13, wherein the plant is a corn plant. 15. The composition according to claim 14, wherein the corn plant is conventional corn, transgenic corn, mutagenic corn, modified corn, sweet corn, or popcorn. 16. Use of a
a plant growth regulator and a fungicide; or a plant growth regulator and an insecticide; or a plant growth regulator and an acaricide; or a plant growth regulator and a fungicide and an insecticide; or a plant growth regulator and an insecticide and an acaricide; or a plant growth regulator and a fungicide and an acaricide for a preparation of a composition. 17. Use of a composition of claim 1, for ensuring high yield per area of a corn crop, together with controlling diseases and parasites that affect corn crops. 18. The use according to claim 17, wherein the parasites are selected from the group consisting of caterpillars (Spodoptera frugiperda, Spodoptera latifacea, Spodoptera eridanea, Spodoptera sp.), leafworms (Mocis latipes), leafhoppers (Deois flavopicta), corn planthoppers (Peregrinus maidis and Dalbulus maidis), corn aphids (Rhopalosiphum maidis), sugarcane borer (Diatraea saccharalis), corn earworms (Helicoverpa zea), pentatomid bugs (Dichelops melacanthus and Dichelops furcatus), neotropical brown stinkbugs (Euschistus heros, Nezara viridula, Piezodorus guildine), leaffooted bugs (Leptoglossus zonatus), squash bug (Phthia pitta), armyworms (Pseudaletia sp.) and mites. 19. The use according to claim 17, wherein the diseases are selected from the group consisting of spot blotch (Exerohilum turcicum), white leaf spot or phaeospheria leaf spot (Phaeosphaeria maydis), diplodia spot (Diplodia macrospora), cercospora leaf spot (Cercospora zea-maydis), anthracnose (Colletotrichum graminicola), polissora rust (Puccinia polysora), tropical rust (Physopella zeae) and common rust (Puccinia sorghi). 20. The use according to claim 17, wherein in that said high yield per area of a corn crop relates to a density greater than 100.000 corn plants per hectare. 21. The use according to claim 17, wherein in that the plant is conventional corn, transgenic corn, mutagenic corn, modified corn, sweet corn, or popcorn. 22. A method for reducing excessive growth of corn plants, comprising the step of an application of an agronomically effective amount of a composition, as defined in claim 1, to the plant, its habitat, progeny or propagation material or combination thereof. 23. A method for strengthening the stem of corn plants, comprising the step of an application of an agronomically effective amount of a composition, as defined in claim 1, to the plant, its habitat, progeny or propagation material or combination thereof. 24. A method for strengthening the root system of corn plants, comprising the step of an application of an agronomically effective amount of a composition, as defined in claim 1, to the plant, its habitat, progeny or propagation material or combination thereof. 25. A method for strengthening the adventitious roots of corn plants, characterized by comprising the application of an agronomically effective amount of a composition, as defined in claim 1, to the plant, its habitat, progeny or propagation material or combination thereof. 26. A method for combating pests in plant crops, wherein agronomically effective amount of a composition, as defined in claim 1, is allowed to act to said pests or their habitat or combination thereof. 27. A method for conferring resistance to pest control in a corn crop, comprising an application of an agronomically effective amount of a composition, as defined in claim 1, to the corn crop. 28. The method according to claim 27, wherein the pests are selected from the group consisting of caterpillars (Spodoptera frugiperda, Spodoptera latifacea, Spodoptera eridanea, Spodoptera sp.), leafworms (Mocis latipes), leafhoppers (Deois flavopicta), corn planthoppers (Peregrinus maidis and Dalbulus maidis), corn aphids (Rhopalosiphum maidis), sugarcane borer (Diatraea saccharalis), corn earworms (Helicoverpa zea), pentatomid bugs (Dichelops melacanthus and Dichelops furcatus), neotropical brown stinkbugs (Euschistus heros, Nezara viridula, Piezodorus guildine), leaffooted bugs (Leptoglossus zonatus), squash bug (Phthia picta), armyworms (Pseudaletia sp.) and mites. 29. A method for identifying a composition that ensures high yield of corn crops, comprising the steps of:
a) applying an agronomically effective amount of a candidate composition to one or more plants; b) sprouting said one or more plants; c) comparing the growth rate of said plants with the growth rate of untreated plants; and d) identifying the candidate composition as a composition that ensures high yield in corn crops and that additionally ensures control of diseases caused by parasites that attack corn plants, wherein said candidate composition comprises a plant growth regulator and fungicide(s), a plant growth regulator and insecticide(s), a regulator and insecticide and fungicide, wherein said high yield is related to a reduction of the excessive growth of corn plants, strengthening of the stem of corn plants, strengthening of the corn plant root system. 30. The method according to claim 29, wherein the candidate composition is applied in an amount ranging from 1 to 2000 g a.i./ha, more preferably from 2.5 to 1000 g a.i./ha. 31. A process for preparing a composition, comprising combining agronomically effective amounts of a
a plant growth regulator and a fungicide; or a plant growth regulator and an insecticide; or a plant growth regulator and an acaricide; or a plant growth regulator and a fungicide and an insecticide; or a plant growth regulator and an insecticide and an acaricide; or a plant growth regulator and a fungicide and an acaricide, as defined in claim 1, and, later, properly mixing them with carriers, adjuvants and/or excipients thereof, such as mineral oil, vegetal oil, silicone adjuvants, non-silicone adjuvants, soybean oil methylate esther, nonylphenol ethoxylate, ethoxylate alkyl esther of phosphoric acid, mixture of paraffin hydrocarbons, cyclic paraffins and saturated and unsaturated aromatics. 32. The process according to claim 31, wherein the plant growth regulator is selected from the group consisting of ethephon, mepiquat chloride, Ca-prohexadione, ancymidol, flurprimidol, chlormequat, dikegulac sodium, mefluidide, uniconazole, paclobutazole, 1-naphthaleneacetic acid (NAA), 1-naphthalene acetamide, carbaryl, 6-benzyladenine (6BA), etichlozate, sulphocarbamide, ethylene, gibberellic acid (GA3), GA47, choline chloride, benzyladenine, cyanamide, cynetine and acybenzolar-S-methyl. 33. The process according to claim 31, wherein the plant growth regulator is ethephon. 34. The process according to claim 31, wherein the fungicide is selected from the group consisting of triazoles, strobilurins, benzimidazoles, acylanilides, carboxanilides, benzamides and pyrazoles. 35. The process according to claim 34, wherein:
the triazoles are selected from the group consisting of: azaconazole, bitertanol, bromuconazole, cyproconazole, difenoconazole, diniconazole, epoxyconazole, etaconazole, fenbuconazole, fluquinconazole, flusilazole, flutriafole, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propiconazole, prothioconazole, simeconazole, tebuconazole, tetraconazole, triadimefon, triadimenol, and triticonazole; the strobilurins are selected from the group consisting of: azoxystrobin, enestrobin, picoxystrobin, piraoxystrobin, pyraclostrobin, pyrametostrobin, kresoxim-methyl, trifloxystrobin, dimoxystrobin, metominostrobin, orysastrobin, famoxadone, fluoxastrobin, fenamidone, and pyribencarb; the benzimidazoles are selected from the group consisting of: benomyl, carbendazim, fuberidazole, thiabendazole, thiophanate, and tiophanato-methyl; the acylanilides are selected from the group consisting of: benalaxyl, furalaxyl, metalaxyl, metalaxyl-M, and mefenoxam; the benzamides are selected from the group consisting of: benodanil, fluopyram, flutolanil, and mepronil; the carboxanilides are selected from the group consisting of: fenfuram, carboxin, oxycarboxin, thifluzamide, bixafen, furametpyr, isopyrazam, penflufen, penthiopyrad, sedaxane, and boscalid; and the pyrazoles are selected from the group consisting of: fipronil and ethiprole. 36. The process according to claim 31, wherein:
the insecticide is selected from the group consisting of anthranilamide, phthalic acid diamide, pyrazole analog, avermectins, benzoylurea, biological agent, bis(thiocarbamate), ketoenol, chloro-cyclodiene, spinosyns, pyridyloxypropyl ether, phenylpyrazole, phenylthiourea, carbamates, neonicotinoids, nicotinoids, organophosphorous, oxadiazine, pyrazole, and pyrethroid. 37. The process according to claim 36, wherein:
the anthranilamide are selected from the group consisting of: chlorantraniliprole and cyantraniliprole; the phthalic acid diamide is flubendiamide; the pyrazole analog is chlorfenapyr; the avermectin is abamectin; the benzoylurea is selected from the group consisting of: chlorfluazuron, diflubenzuron, flufenoxuron, lufenuron, novaluron, eflubenzuron, and triflumuron; the biological agent is selected from the group consisting of: Bacillus thuringiensis, Baculovirus anticarsia, and Metarhizium anisopliae; the bis(thiocarbamate) is cartap hydrochloride; the ketoenol is selected from the group consisting of: spiromesifen, spirodiclofen, spirotetramat; the chlorocyclodiene is endosulfan; the spynosyn is spinosad; the pyridyloxypropyl ether is pyriproxyfen; the phenylpyrazole is ethiprole; the phenylthiourea is diafentiuron; the carbamates are selected from the group consisting of: benfuracarb, carbofuran, carbosulfan, furathiocarb, alanycarb, aldicarb, methomyl, and thiodicarb; the neonicotinoid is selected from the group consisting of: acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, and clothianidin, the nicotinoid is flonicamid; the organophosphorous is selected from the group consisting of: acephate, cadusafos, chlorpyrifos, diazinon, dimethoate, disulfoton, ethion, fenitrothion, fenthion, fentoato, phorate, phosmet, fosthiazate, malathion, methamidophos, methidathion, mevinphos, parathion-methyl, pyridaphenthion, pirimiphos-methyl, profenophos, prothiophos, tebupyrimphos, terbuphos, triazophos, and trichlorphon; the oxadiazine is indoxacarb, the pyrazole is fipronil; and the pyrethroid is selected from the group consisting of: alpha-cypermethrin, betacyfluthrin, beta-cypermethrin, bifenthrin, cyfluthrin, cypermethrin, deltamethrin, esfenvalerate, fenpropatrina, fluvalinate, gamma-cyhalothrin, lambda-cyhalothrin, permethrin, and zeta-cypermethrin. 38. The process according to claim 31, wherein the acaricide is selected from the group consisting of avermectins, bis(arylformamidine), ketoenol, chlorodiphenylsulfone, diacyl-hydrazine, diphenyl oxazoline, dinitrophenol, phenylthiourea, oxazolidinadione, pyrazole, pyridazinone, alkyl sulphite, tetrazine, thiadiazinone, and thiazolidine carboxamide. 39. The process according to claim 38, wherein:
the avermectin is abamectin; the bis(arylformamidine) is amitraz; the ketoenol is selected from the group consisting of: spirodiclofen, spirotetramat, and spiromesifen; the chlorodiphenylsulfone is tetradifon; the diacyl-hydrazine is chromafenozide; the diphenyl oxazoline is etoxazole; the dinitrophenol is dinocap; the phenylthiourea is diafentiuron; the oxazolidinedione is famoxadone; the pyrazole is fenpyroximate; the pyridazinone is pyridaben; the alkyl sulphite is propargite; the tetrazine is clofentezine; the thiadiazinone is buprofezin; and the thiazolidine carboxamide is hexythiazox. 40. The process according to claim 31, wherein the plant growth regulator is present in an amount ranging from about 1 to about 1000 g a.i./ha, more preferably, about 2.5 to about 500 g a.i./ha. 41. The process according to claim 31, wherein the fungicide, insecticide or acaricide is present in an amount ranging from about 10 to about 500 g/L or g/kg of formulated product, more preferably, from about 20 to about 400 g/L or g/kg of formulated product. | 1,600 |
1,056 | 16,202,198 | 1,619 | Methods of treating skin in need of treatment with retinoids are provided, which comprise cleansing the skin with a cleansing composition comprising a low irritation polymeric cleansing agent prior to treating with a retinoid. This improves the activity of the retinoid for treating for example signs of skin aging, acne, or rosacea. | 1. A method for treating skin, comprising in sequence:
(a) cleansing skin in need of treatment for signs of skin aging, acne, or rosacea with a cleansing composition comprising a low molecular weight hydrophobically modified polymer; and (b) topically applying to the skin a leave-on composition comprising a retinoid. 2. The method of claim 1, wherein the low molecular weight hydrophobically modified polymer is a non-crosslinked, linear acrylic copolymer. 3. The method of claim 1, wherein the low molecular weight hydrophobically modified polymer is a non-crosslinked, linear acrylic copolymer derived from at least one first monomeric component selected from the group consisting of (meth)acrylic acid and at least one second monomeric component selected from the group consisting of one or more C1 to C9 alkyl (meth)acrylates, wherein the low molecular weight hydrophobically modified polymer has a number average molecular weight of about 100,000 or less. 4. The method of claim 1, wherein the low molecular weight hydrophobically modified polymer is potassium acrylates copolymer. 5. The method of claim 1, wherein the retinoid is selected from the group consisting of retinol, retinal, retinoic acid, retinyl acetate, and retinyl palmitate. 6. The method of claim 1, wherein the retinoid is retinol. 7. A kit comprising:
(a) a cleansing composition comprising a low molecular weight hydrophobically modified polymer; and (b) a leave-on composition comprising a retinoid. 8. A method of increasing the activity of a retinoid, comprising cleansing skin in need of treatment with retinoids with a cleansing composition comprising a low molecular weight hydrophobically modified polymer prior to contacting the skin with the retinoid. 9. The method of claim 8, wherein the activity of the retinoid is for treating signs of skin aging. 10. The method of claim 8, wherein the activity of the retinoid is for treating signs of acne. 11. The method of claim 8, wherein the activity of the retinoid is for treating signs of rosacea. | Methods of treating skin in need of treatment with retinoids are provided, which comprise cleansing the skin with a cleansing composition comprising a low irritation polymeric cleansing agent prior to treating with a retinoid. This improves the activity of the retinoid for treating for example signs of skin aging, acne, or rosacea.1. A method for treating skin, comprising in sequence:
(a) cleansing skin in need of treatment for signs of skin aging, acne, or rosacea with a cleansing composition comprising a low molecular weight hydrophobically modified polymer; and (b) topically applying to the skin a leave-on composition comprising a retinoid. 2. The method of claim 1, wherein the low molecular weight hydrophobically modified polymer is a non-crosslinked, linear acrylic copolymer. 3. The method of claim 1, wherein the low molecular weight hydrophobically modified polymer is a non-crosslinked, linear acrylic copolymer derived from at least one first monomeric component selected from the group consisting of (meth)acrylic acid and at least one second monomeric component selected from the group consisting of one or more C1 to C9 alkyl (meth)acrylates, wherein the low molecular weight hydrophobically modified polymer has a number average molecular weight of about 100,000 or less. 4. The method of claim 1, wherein the low molecular weight hydrophobically modified polymer is potassium acrylates copolymer. 5. The method of claim 1, wherein the retinoid is selected from the group consisting of retinol, retinal, retinoic acid, retinyl acetate, and retinyl palmitate. 6. The method of claim 1, wherein the retinoid is retinol. 7. A kit comprising:
(a) a cleansing composition comprising a low molecular weight hydrophobically modified polymer; and (b) a leave-on composition comprising a retinoid. 8. A method of increasing the activity of a retinoid, comprising cleansing skin in need of treatment with retinoids with a cleansing composition comprising a low molecular weight hydrophobically modified polymer prior to contacting the skin with the retinoid. 9. The method of claim 8, wherein the activity of the retinoid is for treating signs of skin aging. 10. The method of claim 8, wherein the activity of the retinoid is for treating signs of acne. 11. The method of claim 8, wherein the activity of the retinoid is for treating signs of rosacea. | 1,600 |
1,057 | 14,772,437 | 1,653 | A device for testing the cleanliness of a cannula of a medical instrument can include a flexible guiding member having a first end, a second end, and an outer diameter, and a length extending from the first end to the second end, wherein the first end can be rounded and the length of the guiding member can be free of any cleaning element. The device can further include a sponge element containing a dried extractant configured to extract intracellular ATP and having an outer diameter larger than the outer diameter of the guiding member. The device can also include a coupling member located between the second end of the guiding member and the sponge element, wherein the coupling member can be heat bonded to the sponge element and configured to facilitate detachment of the guiding member from the sponge element. | 1. A device for testing the cleanliness of a cannula of a medical instrument comprising:
a flexible guiding member having a first end, a second end, an outer diameter, and a length extending from the first end to the second end, wherein the first end is rounded and the length of the guiding member is free of any cleaning element; a sponge element containing a dried extractant configured to extract intracellular ATP and having an outer diameter larger than the outer diameter of the guiding member; and a coupling member located between the second end of the guiding member and the sponge element, wherein the coupling member is heat bonded to the sponge element and configured to facilitate detachment of the guiding member from the sponge element. 2. The device of claim 1, wherein the coupling member has a length of less than 10 cm and an outer diameter smaller than the outer diameter of the guiding member. 3. The device of claim 1, wherein the coupling member includes a release mechanism having an outer dimension less than about 2 mm. 4. The device of claim 1, wherein the sponge element is configured to absorb and retain water to activate the extractant in less than about 5 min. 5. The device of claim 1, wherein the extractant comprises at least one of Triton X-100, a quaternary-based detergent, tricholoaceitc acid, and protocatechuic acid. 6. The device of claim 5, wherein the extractant further comprises a buffering agent and a detergent-based preservative. 7. The device of claim 1, wherein the guiding member includes polyoxmethylene and the sponge element includes medical-grade polyurethane. 8. A method of testing a cleanliness of a cannula of a medical instrument using a testing device, wherein the testing device comprises a guiding member having a rounded first end and a second end releasably coupled to a sponge element containing a dried extractant, the steps comprising:
wetting the sponge element in at least one of sterile water and ATP-free water to activate the dried extractant; inserting the first end of the guiding member into a proximal end of the cannula; pushing the guiding member into the cannula to pass the first end of the guiding member through the cannula until the first end of the guiding member exits a distal end of the cannula; pulling at least part of the guiding member out of the distal end of the cannula; contacting an inner surface of the cannula with the activated extractant to lyse a cell located on the inner surface of the cannula; and pulling the second end of the guiding member and the sponge element out of the distal end of the cannula. 9. The method of claim 8, further comprising detaching the sponge element from the guiding member. 10, The method of claim 9, further comprising:
introducing the sponge element into a test swab tube; and exposing the sponge element to a reagent in the test swab tube. 11. The method of claim 10, wherein the reagent is a liquid-stable luciferase/luciferin. 12. The method of claim 10, wherein the sponge element is detached from the guiding member following introduction of the sponge element into the test swab tube. 13. The method of claim 10, further comprising:
inserting the test swab tube into an analysis device; analyzing at least part of the contents of the test swab tube using the analysis device to produce a signal associated with a level of cleanliness of the cannula; and outputting a representation of the signal. 14. The method of claim 13, wherein analyzing includes determining a concentration of at least one of adenosine triphosphate, protein, and ninhydrin. 15. The method of claim 13, further comprising determining whether a subsequent cleaning of the cannula is required based on the representation of the signal. 16. The method of claim 8, wherein the extractant comprises at least one of Triton X-100, a quaternary-based detergent, tricholoaceitc acid, and protocatechuic acid. 17. The method of claim 8, further comprising collecting intracellular ATP on the sponge element. 18. A kit for testing a cleanliness of a cannula of a medical instrument, comprising:
a testing device, comprising:
a flexible guiding member having a first end, a second end, and an outer diameter less than about 2 mm, wherein the first end is rounded and a region extending from the first end to the second end is free of any cleaning element;
a sponge element containing a dried extractant and having an outer diameter of less than about 5 mm, wherein the sponge element is cylindrical; and
a releasable coupling member located between the guiding member and the sponge element, wherein the coupling member is fixedly attached to the second end of the guiding member and the sponge element, and
configured to releasably couple the guiding member he sponge element; and
a test swab tube configured to receive the sponge element uncoupled from the guiding member. 19. The kit of claim 18, further comprising a test swab containing a liquid-stable luciferase/luciferin. 20. The kit of claim 18, wherein the test swab tube is configured for at least partial placement within a hand held device configured to output a representation of the cleanliness of the cannula. | A device for testing the cleanliness of a cannula of a medical instrument can include a flexible guiding member having a first end, a second end, and an outer diameter, and a length extending from the first end to the second end, wherein the first end can be rounded and the length of the guiding member can be free of any cleaning element. The device can further include a sponge element containing a dried extractant configured to extract intracellular ATP and having an outer diameter larger than the outer diameter of the guiding member. The device can also include a coupling member located between the second end of the guiding member and the sponge element, wherein the coupling member can be heat bonded to the sponge element and configured to facilitate detachment of the guiding member from the sponge element.1. A device for testing the cleanliness of a cannula of a medical instrument comprising:
a flexible guiding member having a first end, a second end, an outer diameter, and a length extending from the first end to the second end, wherein the first end is rounded and the length of the guiding member is free of any cleaning element; a sponge element containing a dried extractant configured to extract intracellular ATP and having an outer diameter larger than the outer diameter of the guiding member; and a coupling member located between the second end of the guiding member and the sponge element, wherein the coupling member is heat bonded to the sponge element and configured to facilitate detachment of the guiding member from the sponge element. 2. The device of claim 1, wherein the coupling member has a length of less than 10 cm and an outer diameter smaller than the outer diameter of the guiding member. 3. The device of claim 1, wherein the coupling member includes a release mechanism having an outer dimension less than about 2 mm. 4. The device of claim 1, wherein the sponge element is configured to absorb and retain water to activate the extractant in less than about 5 min. 5. The device of claim 1, wherein the extractant comprises at least one of Triton X-100, a quaternary-based detergent, tricholoaceitc acid, and protocatechuic acid. 6. The device of claim 5, wherein the extractant further comprises a buffering agent and a detergent-based preservative. 7. The device of claim 1, wherein the guiding member includes polyoxmethylene and the sponge element includes medical-grade polyurethane. 8. A method of testing a cleanliness of a cannula of a medical instrument using a testing device, wherein the testing device comprises a guiding member having a rounded first end and a second end releasably coupled to a sponge element containing a dried extractant, the steps comprising:
wetting the sponge element in at least one of sterile water and ATP-free water to activate the dried extractant; inserting the first end of the guiding member into a proximal end of the cannula; pushing the guiding member into the cannula to pass the first end of the guiding member through the cannula until the first end of the guiding member exits a distal end of the cannula; pulling at least part of the guiding member out of the distal end of the cannula; contacting an inner surface of the cannula with the activated extractant to lyse a cell located on the inner surface of the cannula; and pulling the second end of the guiding member and the sponge element out of the distal end of the cannula. 9. The method of claim 8, further comprising detaching the sponge element from the guiding member. 10, The method of claim 9, further comprising:
introducing the sponge element into a test swab tube; and exposing the sponge element to a reagent in the test swab tube. 11. The method of claim 10, wherein the reagent is a liquid-stable luciferase/luciferin. 12. The method of claim 10, wherein the sponge element is detached from the guiding member following introduction of the sponge element into the test swab tube. 13. The method of claim 10, further comprising:
inserting the test swab tube into an analysis device; analyzing at least part of the contents of the test swab tube using the analysis device to produce a signal associated with a level of cleanliness of the cannula; and outputting a representation of the signal. 14. The method of claim 13, wherein analyzing includes determining a concentration of at least one of adenosine triphosphate, protein, and ninhydrin. 15. The method of claim 13, further comprising determining whether a subsequent cleaning of the cannula is required based on the representation of the signal. 16. The method of claim 8, wherein the extractant comprises at least one of Triton X-100, a quaternary-based detergent, tricholoaceitc acid, and protocatechuic acid. 17. The method of claim 8, further comprising collecting intracellular ATP on the sponge element. 18. A kit for testing a cleanliness of a cannula of a medical instrument, comprising:
a testing device, comprising:
a flexible guiding member having a first end, a second end, and an outer diameter less than about 2 mm, wherein the first end is rounded and a region extending from the first end to the second end is free of any cleaning element;
a sponge element containing a dried extractant and having an outer diameter of less than about 5 mm, wherein the sponge element is cylindrical; and
a releasable coupling member located between the guiding member and the sponge element, wherein the coupling member is fixedly attached to the second end of the guiding member and the sponge element, and
configured to releasably couple the guiding member he sponge element; and
a test swab tube configured to receive the sponge element uncoupled from the guiding member. 19. The kit of claim 18, further comprising a test swab containing a liquid-stable luciferase/luciferin. 20. The kit of claim 18, wherein the test swab tube is configured for at least partial placement within a hand held device configured to output a representation of the cleanliness of the cannula. | 1,600 |
1,058 | 15,116,895 | 1,617 | Compositions comprise an amount of cinnamaldehyde that is orally tolerable, thus avoiding an unpleasant mouth feeling, and also tolerable in the gastrointestinal tract. The amount of cinnamaldehyde is supplemented by zinc, and the combination is effective to increase at least one of energy expenditure, sympathetic nervous system activity, or fat oxidation, relative to a composition lacking cinnamaldehyde and zinc but otherwise identical. The composition comprising the combination of cinnamaldehyde and zinc can be used in a method to support weight management or promote weight loss, a method for preventing obesity or overweight, and a method for treating obesity or overweight. In an embodiment, the composition comprising cinnamaldehyde is administered to a human. The composition comprising cinnamaldehyde may be a medicament, a food product or a supplement. | 1. A method for weight maintenance comprising administering to an individual in need thereof a composition comprising cinnamaldehyde and zinc. 2. The method of claim 1 wherein the cinnamaldehyde is present in the composition in an amount that is safe and tolerable to ingest and, in combination with the zinc, effective to increase at least one characteristic selected from the group consisting of energy expenditure, sympathetic nervous system activity, and fat oxidation. 3. The method of claim 1 wherein the composition comprises cinnamon essential oil extract that provides at least a portion of the cinnamaldehyde. 4. The method of claim 1 wherein at least a portion of the cinnamaldehyde is selected from the group consisting of isolated cinnamaldehyde and synthesized cinnamaldehyde. 5. A method for promoting weight loss comprising administering to an individual in need thereof a composition comprising cinnamaldehyde and zinc. 6. A method for preventing obesity or overweight comprising administering to an individual at risk thereof a composition comprising cinnamaldehyde and zinc. 7. Method of claim 1 wherein the individual is obese. 8. (canceled) 9. A composition comprising cinnamaldehyde and zinc, the cinnamaldehyde is present in an amount that is safe and tolerable to ingest and, in combination with the zinc, effective to increase at least one characteristic selected from the group consisting of energy expenditure, sympathetic nervous system activity, and fat oxidation. 10. The composition of claim 9 wherein the composition is a food product in which the cinnamaldehyde is present at a flavouring concentration from 31.87 ppm to 6191 ppm. 11. The composition of claim 9 wherein the composition is a food product in which the cinnamaldehyde:zinc ratio is 1:0.5 to 1:0.005. 12. The composition of claim 9 wherein the composition further comprises an additional ingredient in a therapeutically effective amount to promote weight maintenance or weight loss. 13. Method for promoting weight loss of claim 5 wherein the individual is on a weight loss program. 14. The method of claim 13 wherein the weight loss program is selected from the group consisting of a low-fat diet, a low-carbohydrate diet, a low-calorie diet, a very low-calorie diet, endurance training, strength training, and combinations thereof. 15. The composition of claim 9 wherein the food product comprises a component selected from the group consisting of protein, carbohydrate, fat and combinations thereof. | Compositions comprise an amount of cinnamaldehyde that is orally tolerable, thus avoiding an unpleasant mouth feeling, and also tolerable in the gastrointestinal tract. The amount of cinnamaldehyde is supplemented by zinc, and the combination is effective to increase at least one of energy expenditure, sympathetic nervous system activity, or fat oxidation, relative to a composition lacking cinnamaldehyde and zinc but otherwise identical. The composition comprising the combination of cinnamaldehyde and zinc can be used in a method to support weight management or promote weight loss, a method for preventing obesity or overweight, and a method for treating obesity or overweight. In an embodiment, the composition comprising cinnamaldehyde is administered to a human. The composition comprising cinnamaldehyde may be a medicament, a food product or a supplement.1. A method for weight maintenance comprising administering to an individual in need thereof a composition comprising cinnamaldehyde and zinc. 2. The method of claim 1 wherein the cinnamaldehyde is present in the composition in an amount that is safe and tolerable to ingest and, in combination with the zinc, effective to increase at least one characteristic selected from the group consisting of energy expenditure, sympathetic nervous system activity, and fat oxidation. 3. The method of claim 1 wherein the composition comprises cinnamon essential oil extract that provides at least a portion of the cinnamaldehyde. 4. The method of claim 1 wherein at least a portion of the cinnamaldehyde is selected from the group consisting of isolated cinnamaldehyde and synthesized cinnamaldehyde. 5. A method for promoting weight loss comprising administering to an individual in need thereof a composition comprising cinnamaldehyde and zinc. 6. A method for preventing obesity or overweight comprising administering to an individual at risk thereof a composition comprising cinnamaldehyde and zinc. 7. Method of claim 1 wherein the individual is obese. 8. (canceled) 9. A composition comprising cinnamaldehyde and zinc, the cinnamaldehyde is present in an amount that is safe and tolerable to ingest and, in combination with the zinc, effective to increase at least one characteristic selected from the group consisting of energy expenditure, sympathetic nervous system activity, and fat oxidation. 10. The composition of claim 9 wherein the composition is a food product in which the cinnamaldehyde is present at a flavouring concentration from 31.87 ppm to 6191 ppm. 11. The composition of claim 9 wherein the composition is a food product in which the cinnamaldehyde:zinc ratio is 1:0.5 to 1:0.005. 12. The composition of claim 9 wherein the composition further comprises an additional ingredient in a therapeutically effective amount to promote weight maintenance or weight loss. 13. Method for promoting weight loss of claim 5 wherein the individual is on a weight loss program. 14. The method of claim 13 wherein the weight loss program is selected from the group consisting of a low-fat diet, a low-carbohydrate diet, a low-calorie diet, a very low-calorie diet, endurance training, strength training, and combinations thereof. 15. The composition of claim 9 wherein the food product comprises a component selected from the group consisting of protein, carbohydrate, fat and combinations thereof. | 1,600 |
1,059 | 16,587,818 | 1,613 | The embodiments described herein include methods and formulations for treating anaphylaxis and related acute allergic reactions. The methods and formulations include, but are not limited to, methods and formulations for delivering effective concentrations of levocetirizine and montelukast to a patient in need. The methods and formulations can comprise conventional and/or modified-release elements, providing for drug delivery to the patient. | 1. A method of reducing the duration of anaphylaxis in a patient in need thereof, comprising administering to the patient an effective amount of a combination of levocetirizine and montelukast; wherein the combination is administered at the onset of symptoms. 2. The method of claim 1, wherein the combination is administered in a sequential manner. 3. The method of claim 1, wherein the combination is administered in a substantially simultaneous manner. 4. The method of claim 1, further comprising the administration of an additional active agent. 5. The method of claim 4, wherein the additional active agent is selected from the group consisting of: a histamine H2 receptor antagonist, a beta2-agonist, oxygen, saline, a glucocorticoid, and a H1-anti-histamine. 6. The method of claim 5, wherein the histamine H2 antagonist is ranitidine. 7. The method of claim 5, wherein the histamine H2 antagonist is cimetidine. 8. The method of claim 1, further comprising the administration of a vasoactive agent. 9. The method of claim 8, wherein the vasoactive agent is epinephrine. 10. The method of claim 8, wherein the vasoactive agent is dopamine. 11. The method of claim 1, wherein the combination is administered to the patient by one or more of the routes consisting of enteral, intravenous, intraperitoneal, inhalation, intramuscular, subcutaneous and oral. 12. The method of claim 1, wherein the levocetirizine and montelukast are administered by the same route. | The embodiments described herein include methods and formulations for treating anaphylaxis and related acute allergic reactions. The methods and formulations include, but are not limited to, methods and formulations for delivering effective concentrations of levocetirizine and montelukast to a patient in need. The methods and formulations can comprise conventional and/or modified-release elements, providing for drug delivery to the patient.1. A method of reducing the duration of anaphylaxis in a patient in need thereof, comprising administering to the patient an effective amount of a combination of levocetirizine and montelukast; wherein the combination is administered at the onset of symptoms. 2. The method of claim 1, wherein the combination is administered in a sequential manner. 3. The method of claim 1, wherein the combination is administered in a substantially simultaneous manner. 4. The method of claim 1, further comprising the administration of an additional active agent. 5. The method of claim 4, wherein the additional active agent is selected from the group consisting of: a histamine H2 receptor antagonist, a beta2-agonist, oxygen, saline, a glucocorticoid, and a H1-anti-histamine. 6. The method of claim 5, wherein the histamine H2 antagonist is ranitidine. 7. The method of claim 5, wherein the histamine H2 antagonist is cimetidine. 8. The method of claim 1, further comprising the administration of a vasoactive agent. 9. The method of claim 8, wherein the vasoactive agent is epinephrine. 10. The method of claim 8, wherein the vasoactive agent is dopamine. 11. The method of claim 1, wherein the combination is administered to the patient by one or more of the routes consisting of enteral, intravenous, intraperitoneal, inhalation, intramuscular, subcutaneous and oral. 12. The method of claim 1, wherein the levocetirizine and montelukast are administered by the same route. | 1,600 |
1,060 | 15,392,899 | 1,619 | Provided are compositions suitable for lifting or altering the color of keratinous substrates comprising: (a) at least one alkalizing agent; (b) at least two fatty substances; (c) at least one surfactant; (d) at least one cationic polymer; (e) at least one thickening agent: and (f) at least one solvent. The compositions of the present invention may optionally contain at least one colorant. Also provided are methods of using said compositions. | 1. A hair cosmetic composition for lifting or altering the color of keratinous substrates comprising:
a. at least one alkalizing agent selected from alkali metal carbonates, alkali metal phosphates, organic amines, hydroxide base compounds, and mixtures thereof; b. at least two fatty substances selected from alkanes comprising 6 to 16 carbon atoms, fatty alcohols, esters of fatty acids, esters of fatty alcohol, non-silicone oils, non-silicone waxes, silicones and mixtures thereof; c. at least one surfactant selected from anionic surfactants, nonionic surfactants and mixtures thereof; d. at least one cationic polymer; e. at least one thickening agent; f. at least one solvent selected from water, organic solvents and mixtures thereof; and g. at least one colorant. 2. The hair cosmetic composition of claim 1, wherein the alkalizing agents are selected from organic amines, hydroxide base compounds and mixtures thereof. 3. The hair cosmetic composition of claim 2, wherein the alkalizing agents are selected from ammonium hydroxide, ethanolamine and mixtures thereof. 4. The hair cosmetic composition of claim 2, wherein the alkalizing agents are present in an amount of from about 2% to about 20% by weight, based on the weight of the composition. 5. The hair cosmetic composition of claim 4, wherein the alkalizing agents are present in an amount of from about 2% to about 12% by weight, based on the weight of the composition. 6. The hair cosmetic composition of claim 1, wherein the fatty substances are selected from fatty alcohols, glycol ester, glyceryl ester, glycerol ester, non-silicone oils and mixtures thereof. 7. The hair cosmetic composition of claim 6, wherein the composition does not comprise mineral oil. 8. The hair cosmetic composition of claim 6, wherein the fatty substances comprise hydrogenated vegetable oil and glycol distearate. 9. The hair cosmetic composition of claim 6, wherein the fatty substances are present in an amount of from about 0.5% to about 27% by weight, based on the weight of the composition. 10. The hair cosmetic composition of claim 9, wherein the fatty substances are present in an amount of from about 2% to about 25% by weight, based on the weight of the composition. 11. The hair cosmetic composition of claim 1, wherein the surfactants are selected from sulfonate and/or carboxylic surfactants, alkoxylated derivatives of fatty alcohols, alkoxylated derivatives of alkyl phenols, alkoxylated derivatives of fatty acids, alkoxylated derivatives of fatty acid esters and alkoxylated derivatives of fatty acid amides and mixtures thereof. 12. The hair cosmetic composition of claim 11, wherein the surfactants are present in an amount of from about 1% to about 10% by weight, based on the weight of the composition. 13. The hair color composition of claim 12, wherein the surfactants are present in an amount of from about 1% to about 9% by weight, based on the weight of the composition. 14. The hair cosmetic composition of claim 1, wherein the cationic polymers are selected from polyquaternary ammonium polymers, polyurethane derivatives, quaternary ammonium polymers and mixtures thereof. 15. The hair cosmetic composition of claim 14, wherein the cationic polymers are present in an amount of from about 0.1% to about 5% by weight, based on the weight of the composition. 16. The hair cosmetic composition of claim 15, wherein the cationic polymers are present in an amount of from about 1% to about 3% by weight, based on the weight of the composition. 17. The hair cosmetic composition of claim 1, wherein the thickening agents are selected from a polymeric thickener, a non-polymeric thickener, and mixtures thereof. 18. The hair cosmetic composition of claim 17, wherein the thickening agents are present in an amount of from about 0.1% to about 5% by weight, based on the weight of the composition. 19. The hair cosmetic composition of claim 17, wherein the thickening agents are present in an amount of from about 0.3% to about 3.5% by weight, based on the weight of the composition. 20. The hair cosmetic composition of claim 1, wherein the solvent comprises water. 21. The hair cosmetic composition of claim 20, wherein the solvent is present in an amount of from about 40% to about 65% by weight, based on the weight of the composition. 22. The hair cosmetic composition of claim 1, wherein the composition further comprises at least one antioxidant or reducing agent. 23. The hair cosmetic composition of claim 22, wherein the antioxidant or reducing agents are present in an amount of from about 0.5% to about 3% by weight, based on the weight of the composition. 24. (canceled) 25. A method of lifting or altering the color of keratinous substrates, the method comprising applying onto the keratinous substrate a hair cosmetic composition comprising:
a. at least one alkalizing agent selected from alkali metal carbonates, alkali metal phosphates, organic amines, hydroxide base compounds, and mixtures thereof; b. at least two fatty substances selected from alkanes comprising 6 to 16 carbon atoms, fatty alcohols, esters of fatty acids, esters of fatty alcohol, non-silicone oils, non-silicone waxes, silicones and mixtures thereof; c. at least one surfactant selected from anionic, nonionic surfactants and mixtures thereof; d. at least one cationic polymer; e. at least one thickening agent; f. at least one solvent selected from water, organic solvents and mixtures thereof; and g. at least one colorant. | Provided are compositions suitable for lifting or altering the color of keratinous substrates comprising: (a) at least one alkalizing agent; (b) at least two fatty substances; (c) at least one surfactant; (d) at least one cationic polymer; (e) at least one thickening agent: and (f) at least one solvent. The compositions of the present invention may optionally contain at least one colorant. Also provided are methods of using said compositions.1. A hair cosmetic composition for lifting or altering the color of keratinous substrates comprising:
a. at least one alkalizing agent selected from alkali metal carbonates, alkali metal phosphates, organic amines, hydroxide base compounds, and mixtures thereof; b. at least two fatty substances selected from alkanes comprising 6 to 16 carbon atoms, fatty alcohols, esters of fatty acids, esters of fatty alcohol, non-silicone oils, non-silicone waxes, silicones and mixtures thereof; c. at least one surfactant selected from anionic surfactants, nonionic surfactants and mixtures thereof; d. at least one cationic polymer; e. at least one thickening agent; f. at least one solvent selected from water, organic solvents and mixtures thereof; and g. at least one colorant. 2. The hair cosmetic composition of claim 1, wherein the alkalizing agents are selected from organic amines, hydroxide base compounds and mixtures thereof. 3. The hair cosmetic composition of claim 2, wherein the alkalizing agents are selected from ammonium hydroxide, ethanolamine and mixtures thereof. 4. The hair cosmetic composition of claim 2, wherein the alkalizing agents are present in an amount of from about 2% to about 20% by weight, based on the weight of the composition. 5. The hair cosmetic composition of claim 4, wherein the alkalizing agents are present in an amount of from about 2% to about 12% by weight, based on the weight of the composition. 6. The hair cosmetic composition of claim 1, wherein the fatty substances are selected from fatty alcohols, glycol ester, glyceryl ester, glycerol ester, non-silicone oils and mixtures thereof. 7. The hair cosmetic composition of claim 6, wherein the composition does not comprise mineral oil. 8. The hair cosmetic composition of claim 6, wherein the fatty substances comprise hydrogenated vegetable oil and glycol distearate. 9. The hair cosmetic composition of claim 6, wherein the fatty substances are present in an amount of from about 0.5% to about 27% by weight, based on the weight of the composition. 10. The hair cosmetic composition of claim 9, wherein the fatty substances are present in an amount of from about 2% to about 25% by weight, based on the weight of the composition. 11. The hair cosmetic composition of claim 1, wherein the surfactants are selected from sulfonate and/or carboxylic surfactants, alkoxylated derivatives of fatty alcohols, alkoxylated derivatives of alkyl phenols, alkoxylated derivatives of fatty acids, alkoxylated derivatives of fatty acid esters and alkoxylated derivatives of fatty acid amides and mixtures thereof. 12. The hair cosmetic composition of claim 11, wherein the surfactants are present in an amount of from about 1% to about 10% by weight, based on the weight of the composition. 13. The hair color composition of claim 12, wherein the surfactants are present in an amount of from about 1% to about 9% by weight, based on the weight of the composition. 14. The hair cosmetic composition of claim 1, wherein the cationic polymers are selected from polyquaternary ammonium polymers, polyurethane derivatives, quaternary ammonium polymers and mixtures thereof. 15. The hair cosmetic composition of claim 14, wherein the cationic polymers are present in an amount of from about 0.1% to about 5% by weight, based on the weight of the composition. 16. The hair cosmetic composition of claim 15, wherein the cationic polymers are present in an amount of from about 1% to about 3% by weight, based on the weight of the composition. 17. The hair cosmetic composition of claim 1, wherein the thickening agents are selected from a polymeric thickener, a non-polymeric thickener, and mixtures thereof. 18. The hair cosmetic composition of claim 17, wherein the thickening agents are present in an amount of from about 0.1% to about 5% by weight, based on the weight of the composition. 19. The hair cosmetic composition of claim 17, wherein the thickening agents are present in an amount of from about 0.3% to about 3.5% by weight, based on the weight of the composition. 20. The hair cosmetic composition of claim 1, wherein the solvent comprises water. 21. The hair cosmetic composition of claim 20, wherein the solvent is present in an amount of from about 40% to about 65% by weight, based on the weight of the composition. 22. The hair cosmetic composition of claim 1, wherein the composition further comprises at least one antioxidant or reducing agent. 23. The hair cosmetic composition of claim 22, wherein the antioxidant or reducing agents are present in an amount of from about 0.5% to about 3% by weight, based on the weight of the composition. 24. (canceled) 25. A method of lifting or altering the color of keratinous substrates, the method comprising applying onto the keratinous substrate a hair cosmetic composition comprising:
a. at least one alkalizing agent selected from alkali metal carbonates, alkali metal phosphates, organic amines, hydroxide base compounds, and mixtures thereof; b. at least two fatty substances selected from alkanes comprising 6 to 16 carbon atoms, fatty alcohols, esters of fatty acids, esters of fatty alcohol, non-silicone oils, non-silicone waxes, silicones and mixtures thereof; c. at least one surfactant selected from anionic, nonionic surfactants and mixtures thereof; d. at least one cationic polymer; e. at least one thickening agent; f. at least one solvent selected from water, organic solvents and mixtures thereof; and g. at least one colorant. | 1,600 |
1,061 | 16,254,850 | 1,619 | The invention relates to the use of penflufen for protecting wood and wood-comprising materials against wood-destroying basidiomycetes. | 1. A process for protecting wood and wood-comprising materials against attack or destruction by wood-destroying basidiomycetes, the process comprising treating wood or wood-comprising material with a biocide composition comprising biocides, the biocides comprising:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from the group consisting of copper compounds, sulphonamides, triazoles, imidazoles, benzimidazoles; morpholine derivatives, and pyrethroids, in synergistically effective amounts for improved biocidal efficacy against wood-destroying basidiomycetes greater than a mere additive effect obtainable through the use of the respective biocides alone in the same amounts. 2. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from copper compounds. 3. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from sulphonamides. 4. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from triazoles. 5. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from imidazoles. 6. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from benzimidazoles. 7. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from morpholine derivatives. 8. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from pyrethroids. 9. The process according to claim 1, wherein the at least one further biocidally active compound is selected from the group consisting of prochloraz, fenpropimorph, thiabendazole, tebuconazole, propiconazole, permethrin, etofenprox, bifenthrin, cypermethrin, and copper oxide. 10. The process according to claim 9, wherein the biocides in the composition consist of:
the at least one of penflufen and salts thereof; and the at least one further biocidally active compound selected from the group consisting of prochloraz, fenpropimorph, thiabendazole, tebuconazole, propiconazole, permethrin, etofenprox, bifenthrin, cypermethrin, and copper oxides in synergistically effective amounts. 11. The process according to claim 9, wherein the biocide composition comprises:
at least one solvent or diluent, and 0.001% to 5% by weight of the biocides. 12. The process according to claim 11, wherein:
the basidiomycetes comprise holobasidiomycetes; and the treating comprises treating the wood or wood-comprising material by at least one of painting, spraying, drenching, submersion, and impregnation of the wood or wood-comprising material with the biocide composition. 13. The process according to claim 1, wherein the biocide composition comprises:
at least one solvent or diluent; and the biocides in the composition consist of:
the at least one of penflufen and salts thereof; and
at least one of copper oxide and thiabendazole. 14. The process according to claim 1, wherein the biocides comprise a synergistic mixture of A) and B):
A) at least one of penflufen and salts thereof, and B) one of the following further biocidally active compounds:
prochloraz, at a ratio A:B of 3:7 to 1:9;
fenpropimorph, at a ratio A:B of 7:3 to 1:9;
thiabendazole, at a ratio A:B of 9:1 to 2:3;
tebuconazole, at a ratio A:B of 6:4 to 1:9;
propiconazole, at a ratio A:B of 9:1 to 1:9
permethrin, at a ratio A:B of 7:3 to 4:6;
etofenprox, at a ratio A:B of 7:3 to 1:9
bifenthrin, at a ratio A:B of 4:6 to 1:9;
cypermethrin, at a ratio A:B of 7:3 to 1:9; and
copper oxide, at a ratio A:B of 7:3 to 2:8. 15. The process according to claim 14, wherein the biocide composition comprises:
at least one solvent or diluent, and 0.001% to 5% by weight of the biocides. 16. The process according to claim 14, wherein the biocide composition comprises:
at least one solvent or diluent, and 0.001% to 1% by weight of the biocides. | The invention relates to the use of penflufen for protecting wood and wood-comprising materials against wood-destroying basidiomycetes.1. A process for protecting wood and wood-comprising materials against attack or destruction by wood-destroying basidiomycetes, the process comprising treating wood or wood-comprising material with a biocide composition comprising biocides, the biocides comprising:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from the group consisting of copper compounds, sulphonamides, triazoles, imidazoles, benzimidazoles; morpholine derivatives, and pyrethroids, in synergistically effective amounts for improved biocidal efficacy against wood-destroying basidiomycetes greater than a mere additive effect obtainable through the use of the respective biocides alone in the same amounts. 2. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from copper compounds. 3. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from sulphonamides. 4. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from triazoles. 5. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from imidazoles. 6. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from benzimidazoles. 7. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from morpholine derivatives. 8. The process according to claim 1, wherein the biocides comprise synergistically effective amounts of:
at least one of penflufen and salts thereof; and at least one further biocidally active compound selected from pyrethroids. 9. The process according to claim 1, wherein the at least one further biocidally active compound is selected from the group consisting of prochloraz, fenpropimorph, thiabendazole, tebuconazole, propiconazole, permethrin, etofenprox, bifenthrin, cypermethrin, and copper oxide. 10. The process according to claim 9, wherein the biocides in the composition consist of:
the at least one of penflufen and salts thereof; and the at least one further biocidally active compound selected from the group consisting of prochloraz, fenpropimorph, thiabendazole, tebuconazole, propiconazole, permethrin, etofenprox, bifenthrin, cypermethrin, and copper oxides in synergistically effective amounts. 11. The process according to claim 9, wherein the biocide composition comprises:
at least one solvent or diluent, and 0.001% to 5% by weight of the biocides. 12. The process according to claim 11, wherein:
the basidiomycetes comprise holobasidiomycetes; and the treating comprises treating the wood or wood-comprising material by at least one of painting, spraying, drenching, submersion, and impregnation of the wood or wood-comprising material with the biocide composition. 13. The process according to claim 1, wherein the biocide composition comprises:
at least one solvent or diluent; and the biocides in the composition consist of:
the at least one of penflufen and salts thereof; and
at least one of copper oxide and thiabendazole. 14. The process according to claim 1, wherein the biocides comprise a synergistic mixture of A) and B):
A) at least one of penflufen and salts thereof, and B) one of the following further biocidally active compounds:
prochloraz, at a ratio A:B of 3:7 to 1:9;
fenpropimorph, at a ratio A:B of 7:3 to 1:9;
thiabendazole, at a ratio A:B of 9:1 to 2:3;
tebuconazole, at a ratio A:B of 6:4 to 1:9;
propiconazole, at a ratio A:B of 9:1 to 1:9
permethrin, at a ratio A:B of 7:3 to 4:6;
etofenprox, at a ratio A:B of 7:3 to 1:9
bifenthrin, at a ratio A:B of 4:6 to 1:9;
cypermethrin, at a ratio A:B of 7:3 to 1:9; and
copper oxide, at a ratio A:B of 7:3 to 2:8. 15. The process according to claim 14, wherein the biocide composition comprises:
at least one solvent or diluent, and 0.001% to 5% by weight of the biocides. 16. The process according to claim 14, wherein the biocide composition comprises:
at least one solvent or diluent, and 0.001% to 1% by weight of the biocides. | 1,600 |
1,062 | 15,326,531 | 1,611 | The present invention relates to a pediatric chewable tablet comprising a therapeutically effective amount of an anti-viral agent or pharmaceutical acceptable salt or derivative thereof, in particular Valaciclovir in complex with an ion exchange resin in a specific ratio in order to obtain a palatable and child-friendly product. It also relates to a process for the preparation thereof. | 1. A chewable tablet comprising Valaciclovir or pharmaceutical acceptable salt or derivative thereof in complex with an ion exchange resin, wherein the ratio of Valaciclovir to the ion exchange resin is 1:0.8. 2. The chewable tablet according to claim 1, wherein the ion exchange resin is a cationic acid ion exchange resin. 3. The chewable tablet according to claim 1, further comprising one or more diluents. 4. The chewable tablet according to claim 3, wherein the diluents are selected from dextrates and silicified microcrystalline cellulose. 5. The chewable tablet according to claim 1, wherein the chewable tablet is appropriate for children above the age of 6 years. 6. A process for the preparation of a chewable tablet, as defined in claim 1, comprising the following steps:
Dry blending of Drug: Ion Exchange Resin in the ratio 1:0.8; Kneading the above blend with water in the ratio Drug: Ion Exchange Resin: Water 1:0.8:0.5; Drying of the wet mass at 40° C.; Milling of the Drug Resin Complex until particle size gets less than 250 μm; Dry mixing of the Drug Resin Complex and the excipients of the internal phase; Mixing with the excipients of the external phase; Sifting the powder to eliminate any clumps; Compressing into tablet dosage form. 7. The process according to claim 6, wherein the ion exchange resin is a cationic acid ion exchange resin. 8. The process according to claim 6, wherein the chewable tablet is appropriate for children above the age of 6 years. | The present invention relates to a pediatric chewable tablet comprising a therapeutically effective amount of an anti-viral agent or pharmaceutical acceptable salt or derivative thereof, in particular Valaciclovir in complex with an ion exchange resin in a specific ratio in order to obtain a palatable and child-friendly product. It also relates to a process for the preparation thereof.1. A chewable tablet comprising Valaciclovir or pharmaceutical acceptable salt or derivative thereof in complex with an ion exchange resin, wherein the ratio of Valaciclovir to the ion exchange resin is 1:0.8. 2. The chewable tablet according to claim 1, wherein the ion exchange resin is a cationic acid ion exchange resin. 3. The chewable tablet according to claim 1, further comprising one or more diluents. 4. The chewable tablet according to claim 3, wherein the diluents are selected from dextrates and silicified microcrystalline cellulose. 5. The chewable tablet according to claim 1, wherein the chewable tablet is appropriate for children above the age of 6 years. 6. A process for the preparation of a chewable tablet, as defined in claim 1, comprising the following steps:
Dry blending of Drug: Ion Exchange Resin in the ratio 1:0.8; Kneading the above blend with water in the ratio Drug: Ion Exchange Resin: Water 1:0.8:0.5; Drying of the wet mass at 40° C.; Milling of the Drug Resin Complex until particle size gets less than 250 μm; Dry mixing of the Drug Resin Complex and the excipients of the internal phase; Mixing with the excipients of the external phase; Sifting the powder to eliminate any clumps; Compressing into tablet dosage form. 7. The process according to claim 6, wherein the ion exchange resin is a cationic acid ion exchange resin. 8. The process according to claim 6, wherein the chewable tablet is appropriate for children above the age of 6 years. | 1,600 |
1,063 | 15,838,679 | 1,612 | Described herein are oral care compositions, comprising a peroxydone complex, a dispersant, and a polyvinylpyrrolidone copolymer structure-building agent. | 1. An oral care composition, comprising:
from about 0.01% to about 60% of peroxydone complex, based on a total weight of the oral care composition; from about 0.01% to about 990% of a non-aqueous dispersant, based on the total weight of the oral care composition, and from about 0.01% to about 60% of a polyvinylpyrrolidone copolymer structure-building agent, based on the total weight of the oral care composition, wherein the non-aqueous dispersant comprises a non-aqueous liquid, and the oral care composition includes from about 1% to about 70% of the non-aqueous liquid, based on the total weight of the oral care composition. 2. The oral care composition of claim 1, wherein the peroxydone complex includes a crosslinked polyvinylpyrrolidone complexed with hydrogen peroxide (PVP-H2O2). 3. The oral care composition of claim 2, wherein the polyvinylpyrrolidone copolymer is a polyvinylpyrrolidone vinyl acetate copolymer. 4. The oral care composition of claim 1, wherein the dispersant comprises a liquid poloxamer or a paste poloxamer. 5. The oral care composition of claim 4, wherein the liquid poloxamer comprises polyoxyethylene-polyoxypropylene glycol. 6. The oral care composition of claim 1, wherein the non-aqueous liquid comprises one or more of the group consisting of glycerin monoacetate, triacetin, diethylene glycol diacetate, ethylene glycol diacetate, and propylene glycol diacetate (PGDA). 7. The oral care composition of claim 6, wherein the non-aqueous liquid comprises triacetin, and wherein the oral care composition comprises at least 0.01% triacetin, based on the total weight of the oral care composition. 8. The oral care composition of claim 6, wherein the non-aqueous liquid comprises PGDA and wherein the oral care composition comprises at least 0.01% PGDA, based on the total weight of the oral care composition. 9. The oral care composition of claim 1, wherein the oral care composition comprises an equal or greater amount of non-aqueous liquid to polyvinylpyrrolidone copolymer structure-building agent, such that a mass ratio of the non-aqueous liquid to the polyvinylpyrrolidone copolymer structure-building agent is 50:50 or greater. 10. The oral care composition of claim 1, wherein a mass ratio of the non-aqueous liquid to the polyvinylpyrrolidone copolymer structure-building agent is from about 90 to 50 non-aqueous liquid to about 10 to 50 polyvinylpyrrolidone copolymer structure-building agent. 11. The oral care composition of claim 1, wherein the non-aqueous liquid has a log P value of from about −2.0 to about 2.0 12. The oral care composition of claim 1, wherein a viscosity of the oral care composition is from about 50,000 to about 500,000 cPs. 13. The oral care composition of claim 1, further comprising at least one ingredient from the group consisting of: a whitening agent, a surfactant, an antioxidant, a flavoring, a sweetener, a pH modifiers, an abrasive, an anticalculus agent, a source of fluoride ions, a stannous ion source, a colorant, a dye, and a pigment. 14. The oral care composition of claim 13, wherein the at least one ingredient is orally acceptable. 15. The oral care composition of claim 13, wherein the oral care composition is a dentifrice. 16. A non-aqueous oral care composition, comprising:
from about 0.01% to about 60% of peroxydone complex, based on a total weight of the oral care composition; from about 0.01% to about 60% of a amphiphilic copolymer structure-building agent, based on the total weight of the oral care composition; and from about 0.01% to about 99% of a non-aqueous liquid dispersant, based on the total weight of the oral care composition. 17. The non-aqueous oral care composition of claim 16, wherein the amphiphilic copolymer is a polyvinylpyrrolidone vinyl acetate copolymer. 18. The non-aqueous oral care composition of claim 16, wherein the non-aqueous liquid comprises one or more of the group consisting of glycerin monoacetate, triacetin, diethylene glycol diacetate, ethylene glycol diacetate, and propylene glycol diacetate (PGDA). 19. The non-aqueous oral care composition of claim 18, wherein the non-aqueous liquid comprises triacetin. 20. The non-aqueous oral care composition of claim 18, wherein the non-aqueous liquid comprises PGDA. | Described herein are oral care compositions, comprising a peroxydone complex, a dispersant, and a polyvinylpyrrolidone copolymer structure-building agent.1. An oral care composition, comprising:
from about 0.01% to about 60% of peroxydone complex, based on a total weight of the oral care composition; from about 0.01% to about 990% of a non-aqueous dispersant, based on the total weight of the oral care composition, and from about 0.01% to about 60% of a polyvinylpyrrolidone copolymer structure-building agent, based on the total weight of the oral care composition, wherein the non-aqueous dispersant comprises a non-aqueous liquid, and the oral care composition includes from about 1% to about 70% of the non-aqueous liquid, based on the total weight of the oral care composition. 2. The oral care composition of claim 1, wherein the peroxydone complex includes a crosslinked polyvinylpyrrolidone complexed with hydrogen peroxide (PVP-H2O2). 3. The oral care composition of claim 2, wherein the polyvinylpyrrolidone copolymer is a polyvinylpyrrolidone vinyl acetate copolymer. 4. The oral care composition of claim 1, wherein the dispersant comprises a liquid poloxamer or a paste poloxamer. 5. The oral care composition of claim 4, wherein the liquid poloxamer comprises polyoxyethylene-polyoxypropylene glycol. 6. The oral care composition of claim 1, wherein the non-aqueous liquid comprises one or more of the group consisting of glycerin monoacetate, triacetin, diethylene glycol diacetate, ethylene glycol diacetate, and propylene glycol diacetate (PGDA). 7. The oral care composition of claim 6, wherein the non-aqueous liquid comprises triacetin, and wherein the oral care composition comprises at least 0.01% triacetin, based on the total weight of the oral care composition. 8. The oral care composition of claim 6, wherein the non-aqueous liquid comprises PGDA and wherein the oral care composition comprises at least 0.01% PGDA, based on the total weight of the oral care composition. 9. The oral care composition of claim 1, wherein the oral care composition comprises an equal or greater amount of non-aqueous liquid to polyvinylpyrrolidone copolymer structure-building agent, such that a mass ratio of the non-aqueous liquid to the polyvinylpyrrolidone copolymer structure-building agent is 50:50 or greater. 10. The oral care composition of claim 1, wherein a mass ratio of the non-aqueous liquid to the polyvinylpyrrolidone copolymer structure-building agent is from about 90 to 50 non-aqueous liquid to about 10 to 50 polyvinylpyrrolidone copolymer structure-building agent. 11. The oral care composition of claim 1, wherein the non-aqueous liquid has a log P value of from about −2.0 to about 2.0 12. The oral care composition of claim 1, wherein a viscosity of the oral care composition is from about 50,000 to about 500,000 cPs. 13. The oral care composition of claim 1, further comprising at least one ingredient from the group consisting of: a whitening agent, a surfactant, an antioxidant, a flavoring, a sweetener, a pH modifiers, an abrasive, an anticalculus agent, a source of fluoride ions, a stannous ion source, a colorant, a dye, and a pigment. 14. The oral care composition of claim 13, wherein the at least one ingredient is orally acceptable. 15. The oral care composition of claim 13, wherein the oral care composition is a dentifrice. 16. A non-aqueous oral care composition, comprising:
from about 0.01% to about 60% of peroxydone complex, based on a total weight of the oral care composition; from about 0.01% to about 60% of a amphiphilic copolymer structure-building agent, based on the total weight of the oral care composition; and from about 0.01% to about 99% of a non-aqueous liquid dispersant, based on the total weight of the oral care composition. 17. The non-aqueous oral care composition of claim 16, wherein the amphiphilic copolymer is a polyvinylpyrrolidone vinyl acetate copolymer. 18. The non-aqueous oral care composition of claim 16, wherein the non-aqueous liquid comprises one or more of the group consisting of glycerin monoacetate, triacetin, diethylene glycol diacetate, ethylene glycol diacetate, and propylene glycol diacetate (PGDA). 19. The non-aqueous oral care composition of claim 18, wherein the non-aqueous liquid comprises triacetin. 20. The non-aqueous oral care composition of claim 18, wherein the non-aqueous liquid comprises PGDA. | 1,600 |
1,064 | 16,364,121 | 1,629 | Disclosed herein are methods for treating conditions and/or disorders related to hyperglycemia. In particular, the present invention relates to methods of using (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof and insulin or an insulin analogue in the treatment of mellitus and/or disorders related to diabetes mellitus. | 1. A method of treating a subject having diabetes mellitus and/or disorders related to diabetes mellitus comprising administering to the subject an effective amount of (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof, and insulin or an insulin analogue. 2. The method of claim 1, wherein the insulin analogue is glargine, degludec or detemir. 3. The method of claim 2, wherein the insulin analogue is glargine. 4. The method of claim 1, wherein the (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof is administered to the subject in the amount of 15 to 1,000 mg/day; and the insulin or the insulin analogue is administered to the subject in the amount of 0.05 to 3 units/kg/day. 5. The method of claim 4, wherein the (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof is administered to the subject in the amount of 25 to 800 mg/day; and the insulin or the insulin analogue is administered to the subject in the amount of 0.1 to 2 units/kg/day. 6. The method of claim 1, wherein the (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof is administered to the subject in single or divided doses 2 or 3 times each day; and the insulin or the insulin analogue is administered to the subject in single or divided doses 2 or 3 times each day. 7. A pharmaceutical composition for the treatment of diabetes mellitus and/or disorders related to diabetes mellitus comprising:
(R)-(+)-verapamil or a pharmaceutically acceptable salt thereof; insulin or an insulin analogue; and a pharmaceutically acceptable excipient. 8. The pharmaceutical composition of claim 7, wherein the insulin analogue is glargine, degludec or detemir. 9. The pharmaceutical composition of claim 8, wherein the insulin analogue is glargine. 10. (canceled) | Disclosed herein are methods for treating conditions and/or disorders related to hyperglycemia. In particular, the present invention relates to methods of using (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof and insulin or an insulin analogue in the treatment of mellitus and/or disorders related to diabetes mellitus.1. A method of treating a subject having diabetes mellitus and/or disorders related to diabetes mellitus comprising administering to the subject an effective amount of (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof, and insulin or an insulin analogue. 2. The method of claim 1, wherein the insulin analogue is glargine, degludec or detemir. 3. The method of claim 2, wherein the insulin analogue is glargine. 4. The method of claim 1, wherein the (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof is administered to the subject in the amount of 15 to 1,000 mg/day; and the insulin or the insulin analogue is administered to the subject in the amount of 0.05 to 3 units/kg/day. 5. The method of claim 4, wherein the (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof is administered to the subject in the amount of 25 to 800 mg/day; and the insulin or the insulin analogue is administered to the subject in the amount of 0.1 to 2 units/kg/day. 6. The method of claim 1, wherein the (R)-(+)-verapamil or a pharmaceutically acceptable salt thereof is administered to the subject in single or divided doses 2 or 3 times each day; and the insulin or the insulin analogue is administered to the subject in single or divided doses 2 or 3 times each day. 7. A pharmaceutical composition for the treatment of diabetes mellitus and/or disorders related to diabetes mellitus comprising:
(R)-(+)-verapamil or a pharmaceutically acceptable salt thereof; insulin or an insulin analogue; and a pharmaceutically acceptable excipient. 8. The pharmaceutical composition of claim 7, wherein the insulin analogue is glargine, degludec or detemir. 9. The pharmaceutical composition of claim 8, wherein the insulin analogue is glargine. 10. (canceled) | 1,600 |
1,065 | 15,221,143 | 1,634 | A biomolecule analysis kit includes a reaction container configured to perform an enzymatic reaction, the reaction container including a base portion which has a container-shaped portion and a low-adsorption structural portion which is provided on at least the inner surface of the container-shaped portion, the low-adsorption structural portion having an adsorption rate lower than the base portion at which at least one of a sample which becomes a target of analysis in the enzymatic reaction and a reagent for the enzymatic reaction is adsorbed thereonto, wherein a signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container. | 1. A biomolecule analysis kit comprising:
a reaction container configured to perform an enzymatic reaction, the reaction container including: a base portion which has a container-shaped portion, and a low-adsorption structural portion which is provided on at least the inner surface of the container-shaped portion, the low-adsorption structural portion having an adsorption rate lower than the base portion at which at least one of a sample which becomes a target of analysis in the enzymatic reaction and a reagent for the enzymatic reaction is adsorbed thereonto, wherein a signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container. 2. The biomolecule analysis kit according to claim 1, wherein
the low-adsorption structural portion has a lower adsorption rate than that of the base portion at which the sample is absorbed thereonto, and a background in the signal detection is lower than a case in which the base portion is exposed in the reaction container. 3. The biomolecule analysis kit according to claim 1, wherein
the low-adsorption structural portion has a lower adsorption rate than that of the base portion at which the sample is absorbed thereonto; and a signal intensity in the signal detection is higher than a case in which the base portion is exposed in the reaction container. 4. The biomolecule analysis kit according to claim 1, wherein
the reaction container further has a modified portion which is formed by modifying the surface of the base portion such that the adsorption rate within the inner surface of the container-shaped portions becomes lower than the adsorption rate in the base portion; and each of the container-shaped portions has a bottomed cylindrical shape having an approximately circular opening portion having a diameter of equal to or less than 5 μm. 5. The biomolecule analysis kit according to claim 1, wherein
the reaction container further has a low-adsorption substance layer laminated on the base portion such that the adsorption rate within the inner surface of the container-shaped portions becomes lower than the adsorption rate in the base portion; and each of the container-shaped portions has a bottomed cylindrical shape having an approximately circular opening portion having a diameter of equal to or less than 5 μm. 6. A biomolecule analysis kit comprising:
a reaction container configured to perform an enzymatic reaction, the reaction container including a container-shaped portion configured so that a sample that becomes a target of analysis is supplied to the container-shaped portion and a base portion in which the container-shaped portion is formed, and a reagent configured to be supplied to the reaction container and is used for the enzymatic reaction, wherein the reagent contains an adsorption inhibitor for reducing an adsorption rate of at least one of the sample and the reagent with respect to the base portion; and a signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container. 7. The biomolecule analysis kit according to claim 1, wherein
the enzymatic reaction is an isothermal reaction. 8. The biomolecule analysis kit according to claim 1, wherein
the sample that becomes a target of analysis includes any one of DNA, RNA, miRNA, mRNA, and a protein, and a target substance of analysis is any one of DNA, RNA, miRNA, mRNA, and a protein. 9. The biomolecule analysis kit according to claim 8, wherein
the target substance of analysis is a nucleic acid, and the enzymatic reaction is an invader reaction. 10. The biomolecule analysis kit according to claim 1, wherein
the reagent generates a signal by any one of fluorescence, light emission, pH, light absorption, and electric potential. 11. A biomolecule analysis method using the biomolecule analysis kit according to claim 1. 12. A biomolecule analysis kit comprising:
a reaction container configured to perform an enzymatic reaction, the reaction container including a container-shaped portion configured so that a sample is supplied through a flow channel to the container-shaped portion and a base portion in which the container-shaped portion is formed, and a reagent configured to be supplied to the reaction container and used for the enzymatic reaction, wherein the reagent contains a surfactant for reducing the surface tension of the reagent; and fluorescence or a colorimetric signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container. 13. The biomolecule analysis kit according to claim 6, wherein
the adsorption inhibitor is a surfactant. 14. The biomolecule analysis kit according to claim 13, wherein
the surfactant is a nonionic surfactant. 15. The biomolecule analysis kit according to claim 14, wherein
the nonionic surfactant is Tween 20. 16. The biomolecule analysis kit according to claim 14, wherein
the nonionic surfactant is Triton-100. 17. The biomolecule analysis kit according to claim 13, wherein
the concentration of the surfactant is equal to or greater than 0.0005% and equal to or less than 5%. 18. A biomolecule analysis method comprising:
feeding a reagent into a flow channel in a reaction container which has the flow channel and a plurality of wells such that the plurality of wells is filled with the reagent; and feeding the oil sealing solution into the flow channel and sealing the reagent into the plurality of wells with an oil sealing solution, thereby forming the plurality of wells into a plurality of independent reaction containers for nucleic acid detection, wherein any one of the reagent and the oil sealing solution contains a surfactant. 19. The biomolecule analysis method according to claim 18, further comprising:
a step of filling the plurality of wells with a wash buffer through the flow channel before filling the plurality of wells with the reagent. 20. A biomolecule analysis method using the biomolecule analysis kit according to claim 1, wherein
after a wash buffer is supplied to the container-shaped portion, the reagent is supplied to the container-shaped portion. | A biomolecule analysis kit includes a reaction container configured to perform an enzymatic reaction, the reaction container including a base portion which has a container-shaped portion and a low-adsorption structural portion which is provided on at least the inner surface of the container-shaped portion, the low-adsorption structural portion having an adsorption rate lower than the base portion at which at least one of a sample which becomes a target of analysis in the enzymatic reaction and a reagent for the enzymatic reaction is adsorbed thereonto, wherein a signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container.1. A biomolecule analysis kit comprising:
a reaction container configured to perform an enzymatic reaction, the reaction container including: a base portion which has a container-shaped portion, and a low-adsorption structural portion which is provided on at least the inner surface of the container-shaped portion, the low-adsorption structural portion having an adsorption rate lower than the base portion at which at least one of a sample which becomes a target of analysis in the enzymatic reaction and a reagent for the enzymatic reaction is adsorbed thereonto, wherein a signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container. 2. The biomolecule analysis kit according to claim 1, wherein
the low-adsorption structural portion has a lower adsorption rate than that of the base portion at which the sample is absorbed thereonto, and a background in the signal detection is lower than a case in which the base portion is exposed in the reaction container. 3. The biomolecule analysis kit according to claim 1, wherein
the low-adsorption structural portion has a lower adsorption rate than that of the base portion at which the sample is absorbed thereonto; and a signal intensity in the signal detection is higher than a case in which the base portion is exposed in the reaction container. 4. The biomolecule analysis kit according to claim 1, wherein
the reaction container further has a modified portion which is formed by modifying the surface of the base portion such that the adsorption rate within the inner surface of the container-shaped portions becomes lower than the adsorption rate in the base portion; and each of the container-shaped portions has a bottomed cylindrical shape having an approximately circular opening portion having a diameter of equal to or less than 5 μm. 5. The biomolecule analysis kit according to claim 1, wherein
the reaction container further has a low-adsorption substance layer laminated on the base portion such that the adsorption rate within the inner surface of the container-shaped portions becomes lower than the adsorption rate in the base portion; and each of the container-shaped portions has a bottomed cylindrical shape having an approximately circular opening portion having a diameter of equal to or less than 5 μm. 6. A biomolecule analysis kit comprising:
a reaction container configured to perform an enzymatic reaction, the reaction container including a container-shaped portion configured so that a sample that becomes a target of analysis is supplied to the container-shaped portion and a base portion in which the container-shaped portion is formed, and a reagent configured to be supplied to the reaction container and is used for the enzymatic reaction, wherein the reagent contains an adsorption inhibitor for reducing an adsorption rate of at least one of the sample and the reagent with respect to the base portion; and a signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container. 7. The biomolecule analysis kit according to claim 1, wherein
the enzymatic reaction is an isothermal reaction. 8. The biomolecule analysis kit according to claim 1, wherein
the sample that becomes a target of analysis includes any one of DNA, RNA, miRNA, mRNA, and a protein, and a target substance of analysis is any one of DNA, RNA, miRNA, mRNA, and a protein. 9. The biomolecule analysis kit according to claim 8, wherein
the target substance of analysis is a nucleic acid, and the enzymatic reaction is an invader reaction. 10. The biomolecule analysis kit according to claim 1, wherein
the reagent generates a signal by any one of fluorescence, light emission, pH, light absorption, and electric potential. 11. A biomolecule analysis method using the biomolecule analysis kit according to claim 1. 12. A biomolecule analysis kit comprising:
a reaction container configured to perform an enzymatic reaction, the reaction container including a container-shaped portion configured so that a sample is supplied through a flow channel to the container-shaped portion and a base portion in which the container-shaped portion is formed, and a reagent configured to be supplied to the reaction container and used for the enzymatic reaction, wherein the reagent contains a surfactant for reducing the surface tension of the reagent; and fluorescence or a colorimetric signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container. 13. The biomolecule analysis kit according to claim 6, wherein
the adsorption inhibitor is a surfactant. 14. The biomolecule analysis kit according to claim 13, wherein
the surfactant is a nonionic surfactant. 15. The biomolecule analysis kit according to claim 14, wherein
the nonionic surfactant is Tween 20. 16. The biomolecule analysis kit according to claim 14, wherein
the nonionic surfactant is Triton-100. 17. The biomolecule analysis kit according to claim 13, wherein
the concentration of the surfactant is equal to or greater than 0.0005% and equal to or less than 5%. 18. A biomolecule analysis method comprising:
feeding a reagent into a flow channel in a reaction container which has the flow channel and a plurality of wells such that the plurality of wells is filled with the reagent; and feeding the oil sealing solution into the flow channel and sealing the reagent into the plurality of wells with an oil sealing solution, thereby forming the plurality of wells into a plurality of independent reaction containers for nucleic acid detection, wherein any one of the reagent and the oil sealing solution contains a surfactant. 19. The biomolecule analysis method according to claim 18, further comprising:
a step of filling the plurality of wells with a wash buffer through the flow channel before filling the plurality of wells with the reagent. 20. A biomolecule analysis method using the biomolecule analysis kit according to claim 1, wherein
after a wash buffer is supplied to the container-shaped portion, the reagent is supplied to the container-shaped portion. | 1,600 |
1,066 | 14,512,369 | 1,639 | The invention provides methods, apparatuses, and compositions for high-throughput amplification sequencing of specific target sequences in one or more samples. In some aspects, barcode-tagged polynucleotides are sequenced simultaneously and sample sources are identified on the basis of barcode sequences. In some aspects, sequencing data are used to determine one or more genotypes at one or more loci comprising a causal genetic variant. In some aspects, systems and methods of detecting genetic variation are provided. | 1-60. (canceled) 61. A method of detecting genetic variation in a subject's genome comprising:
(a) providing a plurality of clusters of polynucleotides, wherein (i) each cluster comprises multiple copies of a nucleic acid duplex attached to a support; (ii) each duplex in a cluster comprises a first molecule comprising sequences A-B-G′-D′-C′ from 5′ to 3′ and a second molecule comprising sequences C-D-G-B′-A′ from 5′ to 3′; (iii) sequence A′ is complementary to sequence A, sequence B′ is complementary to sequence B, sequence C′ is complementary to sequence C, sequence D′ is complementary to sequence D, and sequence G′ is complementary to sequence G; (iv) sequence G is a portion of a target polynucleotide sequence from a subject and is different for each of a plurality of clusters; (v) sequence B′ is located 5′ with respect to sequence G in the corresponding target polynucleotide sequence; and (vi) each first molecule comprises a barcode sequence; (b) sequencing sequence G′ by extension of a first primer comprising sequence D to produce an R1 sequence for each cluster; (c) sequencing sequence B′ by extension of a second primer comprising sequence A to produce R2 sequence for each cluster; (d) hybridizing a third primer to sequence C′ and sequencing the barcode sequence by extension of the third primer to produce a barcode sequence for each cluster; (e) performing a first alignment using a first algorithm to align all R1 sequences to a first reference sequence; (f) performing a second alignment using a second algorithm to locally align R1 sequences identified in said first alignment as likely to contain an insertion or deletion with respect to the first reference sequence, to produce a single consensus alignment for each insertion or deletion; (g) performing an R2 alignment by aligning all R2 sequences to a second reference sequence; and (h) determining the presence or absence of a sequence variation identified by steps (e) to (g). 62. The method of claim 61, further comprising transmitting a report with the sequence variation determined in step (h) to a receiver. 63. The method of claim 61, wherein the first reference sequence comprises a reference genome. 64. The method of claim 61, wherein the second reference sequence consists of every sequence B for every different target polynucleotide. 65. The method of claim 61, wherein R2 sequences are aligned independently of R1 sequences. 66. The method of claim 61, further comprising discarding an R1 sequence that aligns to a first position in the first reference sequence that is more than 10,000 base pairs away from a second position in the first reference sequence to which the R2 sequence for the same cluster aligns. 67. The method of claim 61, further comprising deleting a portion of an R1 sequence for a cluster when the portion of R1 sequence to be deleted is identical to at least a portion of sequence B′ for that cluster and sequence G is shorter than the R1 sequence for that cluster. 68. The method of claim 61, further comprising deleting a portion of an R1 sequence for a cluster when the portion of R1 sequence to be deleted is identical to at least a portion of any sequence B′, the portion includes either the 5′ or 3′ nucleotide of R1, and either (i) no R2 sequence was produced for the cluster or (ii) R2 sequence produced is not identical to any sequence B. 69. The method of claim 61, wherein performing the first alignment with a system using the first algorithm takes less time to align all R1 reads than would be taken if the system used the second algorithm to perform the first alignment. 70. The method of claim 61, wherein performing the first alignment with a system using the first algorithm uses less system memory to align all R1 reads than would be used if the system used the second algorithm to perform the first alignment. 71. The method of claim 61, wherein said first algorithm is based on Burrows-Wheeler transform. 72. The method of claim 61, wherein said second algorithm is based on Smith-Waterman algorithm or a hash function. 73. The method of claim 61, wherein R1 and R2 sequences are generated for at least 100 different target polynucleotides. 74. The method of claim 61, wherein each barcode differs from every other barcode in a plurality of different barcodes analyzed in parallel. reaction. 75. The method of claim 61, wherein the barcode sequence is located 5′ from sequence D′. 76. The method of claim 61, further comprising grouping sequences from the clusters based on the barcode sequences. 77. The method of claim 75, further comprising discarding all but one of a plurality of R1 sequences having the same sequence and alignment within a barcode sequence grouping. 78. The method of claim 61, wherein sequences A, B, C, and D are at least 5 nucleotides in length. 79. The method of claim 61, wherein sequence G of every cluster is 1 to 1000 nucleotides in length. 80. The method of claim 61, wherein each probe sequence B of a plurality of clusters is complementary to a sequence comprising a causal genetic variant or a sequence within 200 nucleotides of a causal genetic variant. 81. The method of claim 61, wherein an R1 sequence is produced for at least about 108 clusters in a single reaction. 82. The method of claim 61, wherein presence, absence, or allele ratio of one or more causal genetic variants is determined with an accuracy of at least about 90%. 83. The method of claim 61, wherein the consensus sequence identifies an insertion, a deletion, or an insertion and a deletion in a target polynucleotide with an accuracy of at least about 90%. 84. The method of claim 61, wherein each probe sequence B of a plurality of clusters is complementary to a sequence comprising a non-subject sequence or a sequence within 200 nucleotides of a non-subject sequence. 85. The method of claim 61, wherein the presence or absence of one or more non-subject sequences is determined with an accuracy of at least about 90%. 86. The method of claim 61, further comprising calculating a plurality of probabilities based on the R1 sequences for the subject and including the probabilities in the report, wherein each probability is a probability of the subject or a subject's offspring having or developing a disease or trait. 87. A method of detecting genetic variation in a subject's genome comprising:
(a) providing a plurality of clusters of polynucleotides, wherein (i) each cluster comprises multiple copies of a nucleic acid duplex attached to a support; (ii) each duplex in a cluster comprises a first molecule comprising sequences A-B-G′-D′-C′ from 5′ to 3′ and a second molecule comprising sequences C-D-G-B′-A′ from 5′ to 3′, (iii) sequence A′ is complementary to sequence A, sequence B′ is complementary to sequence B, sequence C′ is complementary to sequence C, sequence D′ is complementary to sequence D, and sequence G′ is complementary to sequence G; (iv) sequence G is a portion of a target polynucleotide sequence from a subject and is different for each of a plurality of clusters; (v) sequence B′ is located 5′ with respect to sequence G in the corresponding target polynucleotide sequence; and (vi) each first molecule comprises a barcode sequence; (b) sequencing sequence G′ by extension of a first primer comprising sequence D to produce an R1 sequence for each cluster; (c) sequencing sequence B′ by extension of a second primer comprising sequence A to produce R2 sequence for each cluster; (d) further comprising hybridizing a third primer to sequence C′ and sequencing the barcode sequence by extension of the third primer to produce a barcode sequence for each cluster (e) performing a first alignment using a first algorithm to align all R1 sequences to a first reference sequence; (f) performing a second alignment using a second algorithm to locally align R1 sequences identified in said first alignment as likely to contain an insertion or deletion with respect to the first reference sequence, to produce a single consensus alignment for each insertion or deletion; (g) performing an R2 alignment by aligning all R2 sequences to a second reference sequence; and (h) calculating a plurality of probabilities based on the R1 sequences for the subject and including the probabilities in a report identifying sequence variation identified by steps (e) to (g), wherein each probability is a probability of the subject or a subject's offspring having or developing a disease or trait. 88. The method of claim 87, further comprising transmitting a report with the sequence variation determined in step (h) to a receiver. 89. The method of claim 87, wherein the first reference sequence comprises a reference genome. 90. The method of claim 87, wherein the second reference sequence consists of every sequence B for every different target polynucleotide. 91. The method of claim 87, wherein R2 sequences are aligned independently of R1 sequences. 92. The method of claim 87, further comprising discarding an R1 sequence that aligns to a first position in the first reference sequence that is more than 10,000 base pairs away from a second position in the first reference sequence to which the R2 sequence for the same cluster aligns. 93. The method of claim 87, further comprising deleting a portion of an R1 sequence for a cluster when the portion of R1 sequence to be deleted is identical to at least a portion of sequence B′ for that cluster and sequence G is shorter than the R1 sequence for that cluster. 94. The method of claim 87, further comprising deleting a portion of an R1 sequence for a cluster when the portion of R1 sequence to be deleted is identical to at least a portion of any sequence B′, the portion includes either the 5′ or 3′ nucleotide of R1, and either (i) no R2 sequence was produced for the cluster or (ii) R2 sequence produced is not identical to any sequence B. 95. The method of claim 87, wherein performing the first alignment with a system using the first algorithm takes less time to align all R1 reads than would be taken if the system used the second algorithm to perform the first alignment. 96. The method of claim 87, wherein performing the first alignment with a system using the first algorithm uses less system memory to align all R1 reads than would be used if the system used the second algorithm to perform the first alignment. 97. The method of claim 87, wherein said first algorithm is based on Burrows-Wheeler transform. 98. The method of claim 87, wherein said second algorithm is based on Smith-Waterman algorithm or a hash function. 99. The method of claim 87, wherein R1 and R2 sequences are generated for at least 100 different target polynucleotides. 100. The method of claim 87, wherein each barcode differs from every other barcode in a plurality of different barcodes analyzed in parallel. 101. The method of claim 87, wherein the barcode sequence is associated with a single sample in a pool of samples sequenced in a single reaction. 102. The method of claim 87, wherein each of a plurality of barcode sequences is uniquely associated with a single sample in a pool of samples sequenced in a single reaction. 103. The method of claim 87, wherein the barcode sequence is located 5′ from sequence D′. 104. A method of detecting genetic variation in a subject's genome comprising:
(a) providing a plurality of clusters of polynucleotides, wherein (i) each cluster comprises multiple copies of a nucleic acid duplex attached to a support; (ii) each duplex in a cluster comprises a first molecule comprising sequences A-B-G′-D′-C′ from 5′ to 3′ and a second molecule comprising sequences C-D-G-B′-A′ from 5′ to 3; (iii) sequence A′ is complementary to sequence A, sequence B′ is complementary to sequence B, sequence C′ is complementary to sequence C, sequence D′ is complementary to sequence D, and sequence G′ is complementary to sequence G; (iv) sequence G is a portion of a target polynucleotide sequence from a subject and is different for each of a plurality of clusters; (v) sequence B′ is located 5′ with respect to sequence G in the corresponding target polynucleotide sequence; and (vi) each first molecule comprises a barcode sequence; (b) sequencing sequence G′ by extension of a first primer comprising sequence D to produce an R1 sequence for each cluster; (c) sequencing sequence B′ by extension of a second primer comprising sequence A to produce R2 sequence for each cluster; (d) hybridizing a third primer to sequence C′ and sequencing the barcode sequence by extension of the third primer to produce a barcode sequence for each cluster; (e) further comprising grouping sequences from the clusters based on the barcode sequences; (f) performing a first alignment using a first algorithm to align all R1 sequences to a first reference sequence; (g) performing a second alignment using a second algorithm to locally align R1 sequences identified in said first alignment as likely to contain an insertion or deletion with respect to the first reference sequence, to produce a single consensus alignment for each insertion or deletion; (h) performing an R2 alignment by aligning all R2 sequences to a second reference sequence; and (i) calculating a plurality of probabilities based on the R1 sequences for the subject and including the probabilities in a report identifying sequence variation identified by steps (f) to (h), wherein each probability is a probability of the subject or a subject's offspring having or developing a disease or trait. 105. A method of detecting genetic variation in a subject's genome comprising:
(a) providing a plurality of clusters of polynucleotides, wherein (i) each cluster comprises multiple copies of a nucleic acid duplex attached to a support; (ii) each duplex in a cluster comprises a first molecule comprising sequences A-B-G′-D′-C′ from 5′ to 3′ and a second molecule comprising sequences C-D-G-B′-A′ from 5′ to 3′; (iii) sequence A′ is complementary to sequence A, sequence B′ is complementary to sequence B, sequence C′ is complementary to sequence C, sequence D′ is complementary to sequence D, and sequence G′ is complementary to sequence G; (iv) sequence G is a portion of a target polynucleotide sequence from a subject and is different for each of a plurality of clusters; (v) sequence B′ is located 5′ with respect to sequence G in the corresponding target polynucleotide sequence; and (vi) each first molecule comprises a barcode sequence; (b) sequencing sequence G′ by extension of a first primer comprising sequence D to produce an R1 sequence for each cluster; (c) sequencing sequence B′ by extension of a second primer comprising sequence A to produce R2 sequence for each cluster; (d) hybridizing a third primer to sequence C′ and sequencing the barcode sequence by extension of the third primer to produce a barcode sequence for each cluster (e) grouping sequences from the clusters based on the barcode sequences; and (f) further comprising discarding all but one of a plurality of R1 sequences having the same sequence and alignment within a barcode sequence grouping; (g) performing a first alignment using a first algorithm to align all R1 sequences to a first reference sequence; (h) performing a second alignment using a second algorithm to locally align R1 sequences identified in said first alignment as likely to contain an insertion or deletion with respect to the first reference sequence, to produce a single consensus alignment for each insertion or deletion; (i) performing an R2 alignment by aligning all R2 sequences to a second reference sequence; and (j) calculating a plurality of probabilities based on the R1 sequences for the subject and including the probabilities in a report identifying sequence variation identified by steps (d) to (f), wherein each probability is a probability of the subject or a subject's offspring having or developing a disease or trait. 106. The method of claim 87, wherein sequences A, B, C, and D are at least 5 nucleotides in length. 107. The method of claim 87, wherein sequence G of every cluster is 1 to 1000 nucleotides in length. 108. The method of claim 87, wherein each probe sequence B of a plurality of clusters is complementary to a sequence comprising a causal genetic variant or a sequence within 200 nucleotides of a causal genetic variant. 109. The method of claim 87, wherein sequence B of one or more of the clusters comprises a sequence selected from the group consisting of SEQ ID NOs:22-121. 110. The method of claim 87, wherein an R1 sequence is produced for at least about 108 clusters in a single reaction. 111. The method of claim 87, wherein presence, absence, or allele ratio of one or more causal genetic variants is determined with an accuracy of at least about 90%. 112. The method of claim 87, wherein the consensus sequence identifies an insertion, a deletion, or an insertion and a deletion in a target polynucleotide with an accuracy of at least about 90%. 113. The method of claim 87, wherein each probe sequence B of a plurality of clusters is complementary to a sequence comprising a nonsubject sequence or a sequence within 200 nucleotides of a non-subject sequence. 114. The method of claim 87, wherein the presence or absence of one or more non-subject sequences is determined with an accuracy of at least about 90%. 115. The method of claim 104 further comprising transmitting the report of step (i) to a receiver. 116. The method of claim 105 further comprising transmitting the report of step (j) to a receiver. 117. A method of detecting genetic variation in a subject's genome comprising:
a. Providing a plurality of fragmented polynucleotides from the subject; b. Ligating a first partially single-stranded adapter to the polynucleotides of step (a), wherein the partially single-stranded adapter has a double-stranded region at one end (sequence U hybridized to complementary sequence U′) and the single-stranded sequence Y that does not hybridize to the target polynucleotide under the hybridization and extension conditions used, and further wherein ligation adds sequence Y to both 5′ ends of the target polynucleotides; c. Hybridizing a plurality of a plurality of different oligonucleotide primers, each having a different target-specific sequence W at the 3′ end and extending the primers to produce an extended oligonucleotide with sequence Y′ (complement of Y) at the 3′ end; d. Optionally, amplifying the extended oligonucleotides with a pair of amplification primers comprising:
i. A first amplification primer comprising sequence X and sequence Y, with sequence Y at the 3′ end for hybridization to sequence Y′;
ii. A second amplification primer comprising sequences V and Z, with Z at the 3′ end for hybridization to sequence Z′ of an extended X-Y primer;
Wherein amplification produces a plurality of extended X-Y oligonucleotides comprising sequences X, Y, W′, and Z′ (5′ to 3′; where W′ is the complement of W, and Z′ is the complement of Z) from the first amplification primer, and a plurality of sequences comprising V, Z, Y′, and X′ (5′ to 3′; where X′ is the complement of X) from the second amplification primer; e. Sequencing a plurality of different target polynucleotides, each contained in a polynucleotide comprising one strand comprising sequences V, Z, W, Y′, and X′ (from 5′ to 3′), and another strand comprising sequences X, Y, W′, Z′, and V′ (from 5′ to 3′), with target polynucleotide sequence located between Z/Y′ and between Z′/Y; f. Determining if the subject has a genetic variation based on the sequencing of step (e). 118. The method of claim 117, wherein each oligonucleotide primer of step (c) further comprises a binding partner at the 5′ end. 119. The method of claim 117, wherein the extended oligonucleotides are amplified. 120. The method of claim 119, wherein the extended oligonucleotides are exponentially amplified. 121. The method of claim 117, wherein sequence Z is common among all oligonucleotide primers. 122. The method of claim 117, wherein sequence W is different for each different oligonucleotide primer, is positioned at the 3′ end of each oligonucleotide primer, and is complementary to a sequence comprising a causal genetic variant or a sequence within 200 nucleotides of a causal genetic variant. 123. The method of claim 117, wherein one or more of sequences V, W, X, Y, and Z are different sequences. 124. The method of claim 117, wherein one or more of sequences V, W, X, Y, and Z comprise 5 or more nucleotides each. 125. The method of claim 117, wherein sequence W of one or more of the plurality of oligonucleotide primers comprises a sequence selected from the group consisting of SEQ ID NOs 22-121. 126. The method of claim 117, wherein the fragmented polynucleotides have a median length between about 200 and about 1000 base pairs. 127. The method of claim 117, wherein sequence Y is positioned at the 3′ end of the adapter. 128. The method of claim 117, wherein the fragmented polynucleotides are treated to produce blunt ends or to have a defined overhang prior to step (a). 129. The method of claim 128, wherein the overhang consists of an adenine. 130. The method of claim 117, further comprising transmitting a report identifying the sequence variation determined in step (f) to a receiver. | The invention provides methods, apparatuses, and compositions for high-throughput amplification sequencing of specific target sequences in one or more samples. In some aspects, barcode-tagged polynucleotides are sequenced simultaneously and sample sources are identified on the basis of barcode sequences. In some aspects, sequencing data are used to determine one or more genotypes at one or more loci comprising a causal genetic variant. In some aspects, systems and methods of detecting genetic variation are provided.1-60. (canceled) 61. A method of detecting genetic variation in a subject's genome comprising:
(a) providing a plurality of clusters of polynucleotides, wherein (i) each cluster comprises multiple copies of a nucleic acid duplex attached to a support; (ii) each duplex in a cluster comprises a first molecule comprising sequences A-B-G′-D′-C′ from 5′ to 3′ and a second molecule comprising sequences C-D-G-B′-A′ from 5′ to 3′; (iii) sequence A′ is complementary to sequence A, sequence B′ is complementary to sequence B, sequence C′ is complementary to sequence C, sequence D′ is complementary to sequence D, and sequence G′ is complementary to sequence G; (iv) sequence G is a portion of a target polynucleotide sequence from a subject and is different for each of a plurality of clusters; (v) sequence B′ is located 5′ with respect to sequence G in the corresponding target polynucleotide sequence; and (vi) each first molecule comprises a barcode sequence; (b) sequencing sequence G′ by extension of a first primer comprising sequence D to produce an R1 sequence for each cluster; (c) sequencing sequence B′ by extension of a second primer comprising sequence A to produce R2 sequence for each cluster; (d) hybridizing a third primer to sequence C′ and sequencing the barcode sequence by extension of the third primer to produce a barcode sequence for each cluster; (e) performing a first alignment using a first algorithm to align all R1 sequences to a first reference sequence; (f) performing a second alignment using a second algorithm to locally align R1 sequences identified in said first alignment as likely to contain an insertion or deletion with respect to the first reference sequence, to produce a single consensus alignment for each insertion or deletion; (g) performing an R2 alignment by aligning all R2 sequences to a second reference sequence; and (h) determining the presence or absence of a sequence variation identified by steps (e) to (g). 62. The method of claim 61, further comprising transmitting a report with the sequence variation determined in step (h) to a receiver. 63. The method of claim 61, wherein the first reference sequence comprises a reference genome. 64. The method of claim 61, wherein the second reference sequence consists of every sequence B for every different target polynucleotide. 65. The method of claim 61, wherein R2 sequences are aligned independently of R1 sequences. 66. The method of claim 61, further comprising discarding an R1 sequence that aligns to a first position in the first reference sequence that is more than 10,000 base pairs away from a second position in the first reference sequence to which the R2 sequence for the same cluster aligns. 67. The method of claim 61, further comprising deleting a portion of an R1 sequence for a cluster when the portion of R1 sequence to be deleted is identical to at least a portion of sequence B′ for that cluster and sequence G is shorter than the R1 sequence for that cluster. 68. The method of claim 61, further comprising deleting a portion of an R1 sequence for a cluster when the portion of R1 sequence to be deleted is identical to at least a portion of any sequence B′, the portion includes either the 5′ or 3′ nucleotide of R1, and either (i) no R2 sequence was produced for the cluster or (ii) R2 sequence produced is not identical to any sequence B. 69. The method of claim 61, wherein performing the first alignment with a system using the first algorithm takes less time to align all R1 reads than would be taken if the system used the second algorithm to perform the first alignment. 70. The method of claim 61, wherein performing the first alignment with a system using the first algorithm uses less system memory to align all R1 reads than would be used if the system used the second algorithm to perform the first alignment. 71. The method of claim 61, wherein said first algorithm is based on Burrows-Wheeler transform. 72. The method of claim 61, wherein said second algorithm is based on Smith-Waterman algorithm or a hash function. 73. The method of claim 61, wherein R1 and R2 sequences are generated for at least 100 different target polynucleotides. 74. The method of claim 61, wherein each barcode differs from every other barcode in a plurality of different barcodes analyzed in parallel. reaction. 75. The method of claim 61, wherein the barcode sequence is located 5′ from sequence D′. 76. The method of claim 61, further comprising grouping sequences from the clusters based on the barcode sequences. 77. The method of claim 75, further comprising discarding all but one of a plurality of R1 sequences having the same sequence and alignment within a barcode sequence grouping. 78. The method of claim 61, wherein sequences A, B, C, and D are at least 5 nucleotides in length. 79. The method of claim 61, wherein sequence G of every cluster is 1 to 1000 nucleotides in length. 80. The method of claim 61, wherein each probe sequence B of a plurality of clusters is complementary to a sequence comprising a causal genetic variant or a sequence within 200 nucleotides of a causal genetic variant. 81. The method of claim 61, wherein an R1 sequence is produced for at least about 108 clusters in a single reaction. 82. The method of claim 61, wherein presence, absence, or allele ratio of one or more causal genetic variants is determined with an accuracy of at least about 90%. 83. The method of claim 61, wherein the consensus sequence identifies an insertion, a deletion, or an insertion and a deletion in a target polynucleotide with an accuracy of at least about 90%. 84. The method of claim 61, wherein each probe sequence B of a plurality of clusters is complementary to a sequence comprising a non-subject sequence or a sequence within 200 nucleotides of a non-subject sequence. 85. The method of claim 61, wherein the presence or absence of one or more non-subject sequences is determined with an accuracy of at least about 90%. 86. The method of claim 61, further comprising calculating a plurality of probabilities based on the R1 sequences for the subject and including the probabilities in the report, wherein each probability is a probability of the subject or a subject's offspring having or developing a disease or trait. 87. A method of detecting genetic variation in a subject's genome comprising:
(a) providing a plurality of clusters of polynucleotides, wherein (i) each cluster comprises multiple copies of a nucleic acid duplex attached to a support; (ii) each duplex in a cluster comprises a first molecule comprising sequences A-B-G′-D′-C′ from 5′ to 3′ and a second molecule comprising sequences C-D-G-B′-A′ from 5′ to 3′, (iii) sequence A′ is complementary to sequence A, sequence B′ is complementary to sequence B, sequence C′ is complementary to sequence C, sequence D′ is complementary to sequence D, and sequence G′ is complementary to sequence G; (iv) sequence G is a portion of a target polynucleotide sequence from a subject and is different for each of a plurality of clusters; (v) sequence B′ is located 5′ with respect to sequence G in the corresponding target polynucleotide sequence; and (vi) each first molecule comprises a barcode sequence; (b) sequencing sequence G′ by extension of a first primer comprising sequence D to produce an R1 sequence for each cluster; (c) sequencing sequence B′ by extension of a second primer comprising sequence A to produce R2 sequence for each cluster; (d) further comprising hybridizing a third primer to sequence C′ and sequencing the barcode sequence by extension of the third primer to produce a barcode sequence for each cluster (e) performing a first alignment using a first algorithm to align all R1 sequences to a first reference sequence; (f) performing a second alignment using a second algorithm to locally align R1 sequences identified in said first alignment as likely to contain an insertion or deletion with respect to the first reference sequence, to produce a single consensus alignment for each insertion or deletion; (g) performing an R2 alignment by aligning all R2 sequences to a second reference sequence; and (h) calculating a plurality of probabilities based on the R1 sequences for the subject and including the probabilities in a report identifying sequence variation identified by steps (e) to (g), wherein each probability is a probability of the subject or a subject's offspring having or developing a disease or trait. 88. The method of claim 87, further comprising transmitting a report with the sequence variation determined in step (h) to a receiver. 89. The method of claim 87, wherein the first reference sequence comprises a reference genome. 90. The method of claim 87, wherein the second reference sequence consists of every sequence B for every different target polynucleotide. 91. The method of claim 87, wherein R2 sequences are aligned independently of R1 sequences. 92. The method of claim 87, further comprising discarding an R1 sequence that aligns to a first position in the first reference sequence that is more than 10,000 base pairs away from a second position in the first reference sequence to which the R2 sequence for the same cluster aligns. 93. The method of claim 87, further comprising deleting a portion of an R1 sequence for a cluster when the portion of R1 sequence to be deleted is identical to at least a portion of sequence B′ for that cluster and sequence G is shorter than the R1 sequence for that cluster. 94. The method of claim 87, further comprising deleting a portion of an R1 sequence for a cluster when the portion of R1 sequence to be deleted is identical to at least a portion of any sequence B′, the portion includes either the 5′ or 3′ nucleotide of R1, and either (i) no R2 sequence was produced for the cluster or (ii) R2 sequence produced is not identical to any sequence B. 95. The method of claim 87, wherein performing the first alignment with a system using the first algorithm takes less time to align all R1 reads than would be taken if the system used the second algorithm to perform the first alignment. 96. The method of claim 87, wherein performing the first alignment with a system using the first algorithm uses less system memory to align all R1 reads than would be used if the system used the second algorithm to perform the first alignment. 97. The method of claim 87, wherein said first algorithm is based on Burrows-Wheeler transform. 98. The method of claim 87, wherein said second algorithm is based on Smith-Waterman algorithm or a hash function. 99. The method of claim 87, wherein R1 and R2 sequences are generated for at least 100 different target polynucleotides. 100. The method of claim 87, wherein each barcode differs from every other barcode in a plurality of different barcodes analyzed in parallel. 101. The method of claim 87, wherein the barcode sequence is associated with a single sample in a pool of samples sequenced in a single reaction. 102. The method of claim 87, wherein each of a plurality of barcode sequences is uniquely associated with a single sample in a pool of samples sequenced in a single reaction. 103. The method of claim 87, wherein the barcode sequence is located 5′ from sequence D′. 104. A method of detecting genetic variation in a subject's genome comprising:
(a) providing a plurality of clusters of polynucleotides, wherein (i) each cluster comprises multiple copies of a nucleic acid duplex attached to a support; (ii) each duplex in a cluster comprises a first molecule comprising sequences A-B-G′-D′-C′ from 5′ to 3′ and a second molecule comprising sequences C-D-G-B′-A′ from 5′ to 3; (iii) sequence A′ is complementary to sequence A, sequence B′ is complementary to sequence B, sequence C′ is complementary to sequence C, sequence D′ is complementary to sequence D, and sequence G′ is complementary to sequence G; (iv) sequence G is a portion of a target polynucleotide sequence from a subject and is different for each of a plurality of clusters; (v) sequence B′ is located 5′ with respect to sequence G in the corresponding target polynucleotide sequence; and (vi) each first molecule comprises a barcode sequence; (b) sequencing sequence G′ by extension of a first primer comprising sequence D to produce an R1 sequence for each cluster; (c) sequencing sequence B′ by extension of a second primer comprising sequence A to produce R2 sequence for each cluster; (d) hybridizing a third primer to sequence C′ and sequencing the barcode sequence by extension of the third primer to produce a barcode sequence for each cluster; (e) further comprising grouping sequences from the clusters based on the barcode sequences; (f) performing a first alignment using a first algorithm to align all R1 sequences to a first reference sequence; (g) performing a second alignment using a second algorithm to locally align R1 sequences identified in said first alignment as likely to contain an insertion or deletion with respect to the first reference sequence, to produce a single consensus alignment for each insertion or deletion; (h) performing an R2 alignment by aligning all R2 sequences to a second reference sequence; and (i) calculating a plurality of probabilities based on the R1 sequences for the subject and including the probabilities in a report identifying sequence variation identified by steps (f) to (h), wherein each probability is a probability of the subject or a subject's offspring having or developing a disease or trait. 105. A method of detecting genetic variation in a subject's genome comprising:
(a) providing a plurality of clusters of polynucleotides, wherein (i) each cluster comprises multiple copies of a nucleic acid duplex attached to a support; (ii) each duplex in a cluster comprises a first molecule comprising sequences A-B-G′-D′-C′ from 5′ to 3′ and a second molecule comprising sequences C-D-G-B′-A′ from 5′ to 3′; (iii) sequence A′ is complementary to sequence A, sequence B′ is complementary to sequence B, sequence C′ is complementary to sequence C, sequence D′ is complementary to sequence D, and sequence G′ is complementary to sequence G; (iv) sequence G is a portion of a target polynucleotide sequence from a subject and is different for each of a plurality of clusters; (v) sequence B′ is located 5′ with respect to sequence G in the corresponding target polynucleotide sequence; and (vi) each first molecule comprises a barcode sequence; (b) sequencing sequence G′ by extension of a first primer comprising sequence D to produce an R1 sequence for each cluster; (c) sequencing sequence B′ by extension of a second primer comprising sequence A to produce R2 sequence for each cluster; (d) hybridizing a third primer to sequence C′ and sequencing the barcode sequence by extension of the third primer to produce a barcode sequence for each cluster (e) grouping sequences from the clusters based on the barcode sequences; and (f) further comprising discarding all but one of a plurality of R1 sequences having the same sequence and alignment within a barcode sequence grouping; (g) performing a first alignment using a first algorithm to align all R1 sequences to a first reference sequence; (h) performing a second alignment using a second algorithm to locally align R1 sequences identified in said first alignment as likely to contain an insertion or deletion with respect to the first reference sequence, to produce a single consensus alignment for each insertion or deletion; (i) performing an R2 alignment by aligning all R2 sequences to a second reference sequence; and (j) calculating a plurality of probabilities based on the R1 sequences for the subject and including the probabilities in a report identifying sequence variation identified by steps (d) to (f), wherein each probability is a probability of the subject or a subject's offspring having or developing a disease or trait. 106. The method of claim 87, wherein sequences A, B, C, and D are at least 5 nucleotides in length. 107. The method of claim 87, wherein sequence G of every cluster is 1 to 1000 nucleotides in length. 108. The method of claim 87, wherein each probe sequence B of a plurality of clusters is complementary to a sequence comprising a causal genetic variant or a sequence within 200 nucleotides of a causal genetic variant. 109. The method of claim 87, wherein sequence B of one or more of the clusters comprises a sequence selected from the group consisting of SEQ ID NOs:22-121. 110. The method of claim 87, wherein an R1 sequence is produced for at least about 108 clusters in a single reaction. 111. The method of claim 87, wherein presence, absence, or allele ratio of one or more causal genetic variants is determined with an accuracy of at least about 90%. 112. The method of claim 87, wherein the consensus sequence identifies an insertion, a deletion, or an insertion and a deletion in a target polynucleotide with an accuracy of at least about 90%. 113. The method of claim 87, wherein each probe sequence B of a plurality of clusters is complementary to a sequence comprising a nonsubject sequence or a sequence within 200 nucleotides of a non-subject sequence. 114. The method of claim 87, wherein the presence or absence of one or more non-subject sequences is determined with an accuracy of at least about 90%. 115. The method of claim 104 further comprising transmitting the report of step (i) to a receiver. 116. The method of claim 105 further comprising transmitting the report of step (j) to a receiver. 117. A method of detecting genetic variation in a subject's genome comprising:
a. Providing a plurality of fragmented polynucleotides from the subject; b. Ligating a first partially single-stranded adapter to the polynucleotides of step (a), wherein the partially single-stranded adapter has a double-stranded region at one end (sequence U hybridized to complementary sequence U′) and the single-stranded sequence Y that does not hybridize to the target polynucleotide under the hybridization and extension conditions used, and further wherein ligation adds sequence Y to both 5′ ends of the target polynucleotides; c. Hybridizing a plurality of a plurality of different oligonucleotide primers, each having a different target-specific sequence W at the 3′ end and extending the primers to produce an extended oligonucleotide with sequence Y′ (complement of Y) at the 3′ end; d. Optionally, amplifying the extended oligonucleotides with a pair of amplification primers comprising:
i. A first amplification primer comprising sequence X and sequence Y, with sequence Y at the 3′ end for hybridization to sequence Y′;
ii. A second amplification primer comprising sequences V and Z, with Z at the 3′ end for hybridization to sequence Z′ of an extended X-Y primer;
Wherein amplification produces a plurality of extended X-Y oligonucleotides comprising sequences X, Y, W′, and Z′ (5′ to 3′; where W′ is the complement of W, and Z′ is the complement of Z) from the first amplification primer, and a plurality of sequences comprising V, Z, Y′, and X′ (5′ to 3′; where X′ is the complement of X) from the second amplification primer; e. Sequencing a plurality of different target polynucleotides, each contained in a polynucleotide comprising one strand comprising sequences V, Z, W, Y′, and X′ (from 5′ to 3′), and another strand comprising sequences X, Y, W′, Z′, and V′ (from 5′ to 3′), with target polynucleotide sequence located between Z/Y′ and between Z′/Y; f. Determining if the subject has a genetic variation based on the sequencing of step (e). 118. The method of claim 117, wherein each oligonucleotide primer of step (c) further comprises a binding partner at the 5′ end. 119. The method of claim 117, wherein the extended oligonucleotides are amplified. 120. The method of claim 119, wherein the extended oligonucleotides are exponentially amplified. 121. The method of claim 117, wherein sequence Z is common among all oligonucleotide primers. 122. The method of claim 117, wherein sequence W is different for each different oligonucleotide primer, is positioned at the 3′ end of each oligonucleotide primer, and is complementary to a sequence comprising a causal genetic variant or a sequence within 200 nucleotides of a causal genetic variant. 123. The method of claim 117, wherein one or more of sequences V, W, X, Y, and Z are different sequences. 124. The method of claim 117, wherein one or more of sequences V, W, X, Y, and Z comprise 5 or more nucleotides each. 125. The method of claim 117, wherein sequence W of one or more of the plurality of oligonucleotide primers comprises a sequence selected from the group consisting of SEQ ID NOs 22-121. 126. The method of claim 117, wherein the fragmented polynucleotides have a median length between about 200 and about 1000 base pairs. 127. The method of claim 117, wherein sequence Y is positioned at the 3′ end of the adapter. 128. The method of claim 117, wherein the fragmented polynucleotides are treated to produce blunt ends or to have a defined overhang prior to step (a). 129. The method of claim 128, wherein the overhang consists of an adenine. 130. The method of claim 117, further comprising transmitting a report identifying the sequence variation determined in step (f) to a receiver. | 1,600 |
1,067 | 15,326,767 | 1,617 | The present invention relates to water dispersible mini-tablets of Enalapril or a pharmaceutically acceptable salt thereof for use in the treatment of hypertension in a pediatric formulation. The pediatric formulation is defined as 0 to 18 years of age. The present invention also provides a method of manufacturing of such dosage form. | 1. A water dispersible minitablet comprising Enalapril or pharmaceutically acceptable salts thereof and one or more pharmaceutically acceptable excipients selected from a disintegrant, a diluent, a lubricant and a glidant, wherein the minitablet is capable of dispersing in water in a period of less than 15 seconds, in accordance with the European Pharmacopoeia {Ph. Eur. 01/2009:2090). 2. The water dispersible minitablet according to claim 1, wherein the disintegrant is Crospovidone, the diluent is Mannitol, the lubricant is Magnesium stearate and the glidant is Talc. 3. The water dispersible minitablet according to claim 1, wherein the concentration of Crospovidone is from 5 to 6% w/w of the total weight of the formulation, the concentration of mannitol is from 85 to 90% w/w of the total weight of the formulation, the concentration of Magnesium stearate is from 0.4 to 0.6% w/w of the total weight of the formulation and the concentration of Talc is from 2 to 4% w/w of the total weight of the formulation. 4. The water dispersible minitablet according to claim 1, wherein Enalapril is Enalapril maleate salt. 5. The water dispersible minitablet according to claim 1, optionally comprising a sweetener, a flavoring or a combination thereof 6. The water dispersible minitablet according to claim 1, wherein the concentration of Enalapril is 0.3125 mg per tablet. 7. The water dispersible minitablet according to claim 1, wherein the size of the minitablet 3 mm in diameter. 8. A process of manufacturing a water dispersible minitablet comprising Enalapril maleate and pharmaceutically acceptable excipients comprising the following steps:
Weighing of raw materials Mixing the pharmaceutically acceptable disintegrant, diluent and glidant with the API until a homogenous powder is formed Lubricating the mixture with the addition of a lubricant Compacting the mixture into minitablets by direct compression Packaging of the minitablets individually in aluminum-aluminum blister foils (Alu-Alu blister packs) 9. The process of manufacturing a minitablet according to claim 8, wherein the disintegrant is Crospovidone, the diluent is Mannitol, the lubricant is Magnesium stearate and the glidant is Talc. 10. A method of treating hypertension in a pediatric population comprising administering the minitablet formulation of claim 1. 11. The method according to claim 10, wherein the pediatric population is from 0 to 18 years of age. 12. The method of claim 10, wherein the minitablet is administered to the patient in need thereof in an amount of not more than 0.5 mg/Kg of weight. | The present invention relates to water dispersible mini-tablets of Enalapril or a pharmaceutically acceptable salt thereof for use in the treatment of hypertension in a pediatric formulation. The pediatric formulation is defined as 0 to 18 years of age. The present invention also provides a method of manufacturing of such dosage form.1. A water dispersible minitablet comprising Enalapril or pharmaceutically acceptable salts thereof and one or more pharmaceutically acceptable excipients selected from a disintegrant, a diluent, a lubricant and a glidant, wherein the minitablet is capable of dispersing in water in a period of less than 15 seconds, in accordance with the European Pharmacopoeia {Ph. Eur. 01/2009:2090). 2. The water dispersible minitablet according to claim 1, wherein the disintegrant is Crospovidone, the diluent is Mannitol, the lubricant is Magnesium stearate and the glidant is Talc. 3. The water dispersible minitablet according to claim 1, wherein the concentration of Crospovidone is from 5 to 6% w/w of the total weight of the formulation, the concentration of mannitol is from 85 to 90% w/w of the total weight of the formulation, the concentration of Magnesium stearate is from 0.4 to 0.6% w/w of the total weight of the formulation and the concentration of Talc is from 2 to 4% w/w of the total weight of the formulation. 4. The water dispersible minitablet according to claim 1, wherein Enalapril is Enalapril maleate salt. 5. The water dispersible minitablet according to claim 1, optionally comprising a sweetener, a flavoring or a combination thereof 6. The water dispersible minitablet according to claim 1, wherein the concentration of Enalapril is 0.3125 mg per tablet. 7. The water dispersible minitablet according to claim 1, wherein the size of the minitablet 3 mm in diameter. 8. A process of manufacturing a water dispersible minitablet comprising Enalapril maleate and pharmaceutically acceptable excipients comprising the following steps:
Weighing of raw materials Mixing the pharmaceutically acceptable disintegrant, diluent and glidant with the API until a homogenous powder is formed Lubricating the mixture with the addition of a lubricant Compacting the mixture into minitablets by direct compression Packaging of the minitablets individually in aluminum-aluminum blister foils (Alu-Alu blister packs) 9. The process of manufacturing a minitablet according to claim 8, wherein the disintegrant is Crospovidone, the diluent is Mannitol, the lubricant is Magnesium stearate and the glidant is Talc. 10. A method of treating hypertension in a pediatric population comprising administering the minitablet formulation of claim 1. 11. The method according to claim 10, wherein the pediatric population is from 0 to 18 years of age. 12. The method of claim 10, wherein the minitablet is administered to the patient in need thereof in an amount of not more than 0.5 mg/Kg of weight. | 1,600 |
1,068 | 15,529,656 | 1,612 | An oral care mouth rinse composition containing at least one ionic tin source and polyvinylpyrrolidone wherein the polyvinylpyrrolidone is present in an amount of from 1.5 to 4 weight % based on the total weight of the oral care mouth rinse composition is provided. | 1. An oral care mouth rinse composition comprising
(i) at least one source of ionic tin and (ii) polyvinylpyrrolidone
wherein the polyvinylpyrrolidone is present in an amount of from 1.5 to 4 weight %, based on the total weight of the oral care mouth rinse composition. 2. The oral care mouth rinse composition according to claim 1 wherein the polyvinylpyrrolidone is present in an amount of from 2.20 to 3.00 weight % based on the total weight of the oral care mouth rinse composition. 3. The oral care mouth rinse composition according to claim 1 wherein the polyvinylpyrrolidone is present in an amount of from 2.20 to 2.50 weight % based on the total weight of the oral care mouth rinse composition. 4. The oral care mouth rinse composition of claim 1 wherein the polyvinylpyrrolidone is present in an amount of from 2.30 to 2.45 weight ° X° based on the total weight of the oral care mouth rinse composition. 5. The oral care mouth rinse composition of claim 1 wherein the composition comprises at least one stannous ion source, at least one stannic ion source or a combination thereof. 6. The oral care mouth rinse composition of claim 1 wherein the composition comprises at least one stannous ion source. 7. The oral care mouth rinse composition of claim 1 wherein the at least one ionic tin source is selected from the group comprising stannous fluoride, stannous chloride, stannic fluoride, stannic chloride, stannic acetate, stannous acetate and combinations thereof. 8. The oral care mouth rinse composition of claim 1 wherein the at least one ionic tin source is selected from the group comprising stannous fluoride, stannous chloride, stannous acetate and combinations thereof. 9. The oral care mouth rinse composition of claim 1 wherein the concentration of ionic tin is from 0.01 to 0.10 weight % based on the total weight of the oral care mouth rinse composition. 10. The oral care mouth rinse composition of claim 1 wherein the concentration of ionic tin is from 0.02 to 0.08 weight % based on the total weight of the oral care mouth rinse composition. 11. The oral care mouth rinse composition of claim 1 wherein the concentration of ionic tin is from 0.03 to 0.06 weight % based on the total weight of the oral care mouth rinse composition. 12. The oral care mouth rinse composition of claim 1 wherein the concentration of ionic tin is from 0.035 to 0.045 weight % based on the total weight of the oral care mouth rinse composition. 13. The oral care mouth rinse composition of claim 1 wherein the at least one-source of ionic tin comprises stannous fluoride. 14. The oral care mouth rinse composition of claim 1 further comprising a fluoride ion source which is not a tin salt, optionally in an amount of from 0.010 to 0.50 weight % based on the total weight of the composition. 15. The oral care mouth rinse composition of claim 1 further comprising a fluoride ion source selected from sodium fluoride, potassium fluoride, sodium monofluorophosphate, sodium fluorosilicate, ammonium fluorosilicate, amine fluoride, ammonium fluoride and combinations thereof, optionally in an amount corresponding to 0.01 to 0.15 weight % of fluoride based on the total weight of the composition. 16. The oral care mouth rinse composition of claim 1 comprising amine fluoride, sodium fluoride or a combination thereof. 17. The oral care mouth rinse composition of claim 1 comprising 0.02 to 0.08 weight % stannous fluoride and 0.010 to 0.50 weight % amine fluoride. 18. The oral care mouth rinse composition of claim 1 comprising 50 to 1500 ppm fluoride. 19. The oral care mouth rinse composition of claim 1 comprising polyvinylpyrrolidone having a molar mass of from 10,000 to 300,000 Daltons. 20. A method to
(i) reduce or inhibit formation of dental caries, (ii) reduce, repair or inhibit pre-carious lesions of the enamel, (iii) reduce or inhibit demineralization and promote remineralization of the teeth (iv) reduce hypersensitivity of the teeth, (v) reduce or inhibit gingivitis, (vi) promote healing of sores or cuts in the oral cavity, (vii) reduce levels of acid producing bacteria, (viii) reduce or inhibit microbial biofilm formation in the oral cavity, (ix) reduce or inhibit plaque formation in the oral cavity, (x) promote systemic health, or (xi) clean teeth and oral cavity,
comprising applying an effective amount of an oral care mouth rinse composition according to claim 1 to the oral cavity of a subject in need thereof. 21. An oral care mouth rinse composition according to claim 1 for use in a method to
(i) reduce or inhibit formation of dental caries,
(ii) reduce, repair or inhibit pre-carious lesions of the enamel,
(iii) reduce or inhibit demineralization and promote remineralization of the teeth,
(iv) reduce hypersensitivity of the teeth,
(v) reduce or inhibit gingivitis,
(vi) promote healing of sores or cuts in the oral cavity,
(vii) reduce levels of acid producing bacteria,
(viii) reduce or inhibit microbial biofilm formation in the oral cavity,
(ix) reduce or inhibit plaque formation in the oral cavity,
(x) promote systemic health, or
(xi) clean teeth and oral cavity. 22. (canceled) 23. A method of reducing staining of surfaces in the oral cavity resulting from use of an oral care mouth rinse comprising at least one source of ionic tin, wherein the method comprises formulating the oral care mouth rinse composition to comprise polyvinylpyrrolidone in an amount of from 2.20 to 3.00 weight % based on the total weight of the oral care mouth rinse composition. 24. The method of claim 23 wherein the method comprises formulating the oral care mouth rinse composition to comprise from 2.30 to 2.45 weight % polyvinylpyrrolidone. 25. The method of claim 23 wherein the concentration of ionic tin in the oral care mouth rinse composition is from 0.02 to 0.08 weight % based on the total weight of the oral care mouth rinse composition. | An oral care mouth rinse composition containing at least one ionic tin source and polyvinylpyrrolidone wherein the polyvinylpyrrolidone is present in an amount of from 1.5 to 4 weight % based on the total weight of the oral care mouth rinse composition is provided.1. An oral care mouth rinse composition comprising
(i) at least one source of ionic tin and (ii) polyvinylpyrrolidone
wherein the polyvinylpyrrolidone is present in an amount of from 1.5 to 4 weight %, based on the total weight of the oral care mouth rinse composition. 2. The oral care mouth rinse composition according to claim 1 wherein the polyvinylpyrrolidone is present in an amount of from 2.20 to 3.00 weight % based on the total weight of the oral care mouth rinse composition. 3. The oral care mouth rinse composition according to claim 1 wherein the polyvinylpyrrolidone is present in an amount of from 2.20 to 2.50 weight % based on the total weight of the oral care mouth rinse composition. 4. The oral care mouth rinse composition of claim 1 wherein the polyvinylpyrrolidone is present in an amount of from 2.30 to 2.45 weight ° X° based on the total weight of the oral care mouth rinse composition. 5. The oral care mouth rinse composition of claim 1 wherein the composition comprises at least one stannous ion source, at least one stannic ion source or a combination thereof. 6. The oral care mouth rinse composition of claim 1 wherein the composition comprises at least one stannous ion source. 7. The oral care mouth rinse composition of claim 1 wherein the at least one ionic tin source is selected from the group comprising stannous fluoride, stannous chloride, stannic fluoride, stannic chloride, stannic acetate, stannous acetate and combinations thereof. 8. The oral care mouth rinse composition of claim 1 wherein the at least one ionic tin source is selected from the group comprising stannous fluoride, stannous chloride, stannous acetate and combinations thereof. 9. The oral care mouth rinse composition of claim 1 wherein the concentration of ionic tin is from 0.01 to 0.10 weight % based on the total weight of the oral care mouth rinse composition. 10. The oral care mouth rinse composition of claim 1 wherein the concentration of ionic tin is from 0.02 to 0.08 weight % based on the total weight of the oral care mouth rinse composition. 11. The oral care mouth rinse composition of claim 1 wherein the concentration of ionic tin is from 0.03 to 0.06 weight % based on the total weight of the oral care mouth rinse composition. 12. The oral care mouth rinse composition of claim 1 wherein the concentration of ionic tin is from 0.035 to 0.045 weight % based on the total weight of the oral care mouth rinse composition. 13. The oral care mouth rinse composition of claim 1 wherein the at least one-source of ionic tin comprises stannous fluoride. 14. The oral care mouth rinse composition of claim 1 further comprising a fluoride ion source which is not a tin salt, optionally in an amount of from 0.010 to 0.50 weight % based on the total weight of the composition. 15. The oral care mouth rinse composition of claim 1 further comprising a fluoride ion source selected from sodium fluoride, potassium fluoride, sodium monofluorophosphate, sodium fluorosilicate, ammonium fluorosilicate, amine fluoride, ammonium fluoride and combinations thereof, optionally in an amount corresponding to 0.01 to 0.15 weight % of fluoride based on the total weight of the composition. 16. The oral care mouth rinse composition of claim 1 comprising amine fluoride, sodium fluoride or a combination thereof. 17. The oral care mouth rinse composition of claim 1 comprising 0.02 to 0.08 weight % stannous fluoride and 0.010 to 0.50 weight % amine fluoride. 18. The oral care mouth rinse composition of claim 1 comprising 50 to 1500 ppm fluoride. 19. The oral care mouth rinse composition of claim 1 comprising polyvinylpyrrolidone having a molar mass of from 10,000 to 300,000 Daltons. 20. A method to
(i) reduce or inhibit formation of dental caries, (ii) reduce, repair or inhibit pre-carious lesions of the enamel, (iii) reduce or inhibit demineralization and promote remineralization of the teeth (iv) reduce hypersensitivity of the teeth, (v) reduce or inhibit gingivitis, (vi) promote healing of sores or cuts in the oral cavity, (vii) reduce levels of acid producing bacteria, (viii) reduce or inhibit microbial biofilm formation in the oral cavity, (ix) reduce or inhibit plaque formation in the oral cavity, (x) promote systemic health, or (xi) clean teeth and oral cavity,
comprising applying an effective amount of an oral care mouth rinse composition according to claim 1 to the oral cavity of a subject in need thereof. 21. An oral care mouth rinse composition according to claim 1 for use in a method to
(i) reduce or inhibit formation of dental caries,
(ii) reduce, repair or inhibit pre-carious lesions of the enamel,
(iii) reduce or inhibit demineralization and promote remineralization of the teeth,
(iv) reduce hypersensitivity of the teeth,
(v) reduce or inhibit gingivitis,
(vi) promote healing of sores or cuts in the oral cavity,
(vii) reduce levels of acid producing bacteria,
(viii) reduce or inhibit microbial biofilm formation in the oral cavity,
(ix) reduce or inhibit plaque formation in the oral cavity,
(x) promote systemic health, or
(xi) clean teeth and oral cavity. 22. (canceled) 23. A method of reducing staining of surfaces in the oral cavity resulting from use of an oral care mouth rinse comprising at least one source of ionic tin, wherein the method comprises formulating the oral care mouth rinse composition to comprise polyvinylpyrrolidone in an amount of from 2.20 to 3.00 weight % based on the total weight of the oral care mouth rinse composition. 24. The method of claim 23 wherein the method comprises formulating the oral care mouth rinse composition to comprise from 2.30 to 2.45 weight % polyvinylpyrrolidone. 25. The method of claim 23 wherein the concentration of ionic tin in the oral care mouth rinse composition is from 0.02 to 0.08 weight % based on the total weight of the oral care mouth rinse composition. | 1,600 |
1,069 | 15,174,540 | 1,653 | Methods and materials for treating bacterial vaginosis (“BV”) are provided. Cervicovaginal secretions (“CVS”) from a woman with vaginal microbiota dominated (>50%) by one of the species of lactobacillus typically found in the human vagina, e.g. Lactobacillus crispatus, L. iners, L. gasseri, L jensenii , is transplanted to women with BV as a method for restoring beneficial vaginal microbial communities and/or increasing resistance to sexually transmitted disease. Efficacy can be enhanced, or the properties of the endogenous CVS improved, through administration of an acidifying agent such as lactic acid. The examples demonstrate the role of healthy CVS in disease resistance, and the effect of pH on CVS properties. The examples also describe the collection and transplantation of healthy beneficial CVS into women at risk for, or after treatment for, BV. | 1. An isolated cervicovaginal secretion comprising vaginal microbiota, wherein at least 50% of the vaginal microbiota is one species typically found in the human vagina, e.g. Lactobacillus crispatus, Lactobacillus iners, Lactobacillus gasseri, or Lactobacillus jensenii, having no evidence of sexually transmitted disease or bacterial vaginosis, an acidic pH less than 4.0. 2. The isolated cervicovaginal secretion of claim 1 which has been sterile filtered and pH adjusted. 3. The sterile filtered and pH adjusted cervicovaginal secretion of claim 2 containing Lactobacillus bacteria isolated, and optionally cultured in vitro. 4. The isolated cervicovaginal secretion of claim 1 packaged into a dosage unit or applicator for administration to a human woman. 5. The isolated cervicovaginal secretion of claim 1 that has been frozen, optionally in combination with a cryoprotectant (i.e. glycerol), and thawed for administration to a human woman. 6. The isolated cervicovaginal secretion of claim 1 that has been mixed with a diluent containing up to 1.5% lactic acid, pH <4.0, for administration to a human woman. 7. The isolated cervicovaginal secretion of claim 1 which has been spray dried or lyophilized and formulated for administration to the vagina, optionally in combination with a resuspending or dissolution agent. 8. A method for treating bacterial vaginosis comprising administering to the vagina an isolated cervicovaginal secretion comprising vaginal microbiota, wherein at least 50% of the vaginal microbiota is one species typically found in the human vagina, e.g. Lactobacillus crispatus, Lactobacillus iners, Lactobacillus gasseri, or Lactobacillus jensenii, having no evidence of sexually transmitted disease or bacterial vaginosis, and an acidic pH less than 4.0, as defined by claim 1. 9. The method of claim 8 further comprising subsequent daily vaginal administration of one to five mls of a 1-1.5% lactic acid gel (pH <4.0) for up to one week before and/or after transplantation of the isolated cervicovaginal secretion. 10. The method of claim 8 wherein the bacterial vaginosis has been treated with antibiotics prior to administration of the cervicovaginal secretion. 11. A method for enhancing resistance to sexually transmitted disease comprising administering to the vagina an isolated cervicovaginal secretion comprising vaginal microbiota, wherein at least 50% of the vaginal microbiota is one species typically found in the human vagina, e.g. Lactobacillus crispatus, Lactobacillus iners, Lactobacillus gasseri, or Lactobacillus jensenii, having no evidence of sexually transmitted disease or bacterial vaginosis, and an acidic pH less than 4.0, as defined by claim 1. 12. The method of claim 11 further comprising administering one to five mls of a 1 to 1.5% food acid gel (pH less than 4.0) daily for up to 1 week before and/or after transplantation of an isolated cervicovaginal secretion. | Methods and materials for treating bacterial vaginosis (“BV”) are provided. Cervicovaginal secretions (“CVS”) from a woman with vaginal microbiota dominated (>50%) by one of the species of lactobacillus typically found in the human vagina, e.g. Lactobacillus crispatus, L. iners, L. gasseri, L jensenii , is transplanted to women with BV as a method for restoring beneficial vaginal microbial communities and/or increasing resistance to sexually transmitted disease. Efficacy can be enhanced, or the properties of the endogenous CVS improved, through administration of an acidifying agent such as lactic acid. The examples demonstrate the role of healthy CVS in disease resistance, and the effect of pH on CVS properties. The examples also describe the collection and transplantation of healthy beneficial CVS into women at risk for, or after treatment for, BV.1. An isolated cervicovaginal secretion comprising vaginal microbiota, wherein at least 50% of the vaginal microbiota is one species typically found in the human vagina, e.g. Lactobacillus crispatus, Lactobacillus iners, Lactobacillus gasseri, or Lactobacillus jensenii, having no evidence of sexually transmitted disease or bacterial vaginosis, an acidic pH less than 4.0. 2. The isolated cervicovaginal secretion of claim 1 which has been sterile filtered and pH adjusted. 3. The sterile filtered and pH adjusted cervicovaginal secretion of claim 2 containing Lactobacillus bacteria isolated, and optionally cultured in vitro. 4. The isolated cervicovaginal secretion of claim 1 packaged into a dosage unit or applicator for administration to a human woman. 5. The isolated cervicovaginal secretion of claim 1 that has been frozen, optionally in combination with a cryoprotectant (i.e. glycerol), and thawed for administration to a human woman. 6. The isolated cervicovaginal secretion of claim 1 that has been mixed with a diluent containing up to 1.5% lactic acid, pH <4.0, for administration to a human woman. 7. The isolated cervicovaginal secretion of claim 1 which has been spray dried or lyophilized and formulated for administration to the vagina, optionally in combination with a resuspending or dissolution agent. 8. A method for treating bacterial vaginosis comprising administering to the vagina an isolated cervicovaginal secretion comprising vaginal microbiota, wherein at least 50% of the vaginal microbiota is one species typically found in the human vagina, e.g. Lactobacillus crispatus, Lactobacillus iners, Lactobacillus gasseri, or Lactobacillus jensenii, having no evidence of sexually transmitted disease or bacterial vaginosis, and an acidic pH less than 4.0, as defined by claim 1. 9. The method of claim 8 further comprising subsequent daily vaginal administration of one to five mls of a 1-1.5% lactic acid gel (pH <4.0) for up to one week before and/or after transplantation of the isolated cervicovaginal secretion. 10. The method of claim 8 wherein the bacterial vaginosis has been treated with antibiotics prior to administration of the cervicovaginal secretion. 11. A method for enhancing resistance to sexually transmitted disease comprising administering to the vagina an isolated cervicovaginal secretion comprising vaginal microbiota, wherein at least 50% of the vaginal microbiota is one species typically found in the human vagina, e.g. Lactobacillus crispatus, Lactobacillus iners, Lactobacillus gasseri, or Lactobacillus jensenii, having no evidence of sexually transmitted disease or bacterial vaginosis, and an acidic pH less than 4.0, as defined by claim 1. 12. The method of claim 11 further comprising administering one to five mls of a 1 to 1.5% food acid gel (pH less than 4.0) daily for up to 1 week before and/or after transplantation of an isolated cervicovaginal secretion. | 1,600 |
1,070 | 15,236,807 | 1,654 | A bone gel composition consists of cortical bone. The cortical bone is made from cut pieces freeze-dried then ground into particles and demineralized then freeze-dried. A volume of the particles is placed in a solution of sterile water to create a mixture, the water volume being at least twice the particle volume, the mixture is autoclaved under heat and pressure to form a gelatin, the resulting bone gel is formed into sheets having a thickness (t). | 1. A bone gel composition consisting of cortical bone consists of cortical bone made from cut pieces freeze dried then ground into particles and demineralized then freeze-dried, wherein a volume of the particles is placed in a solution of sterile water to create a mixture, the water volume being at least twice the volume of the particles, the mixture is autoclaved under heat and pressure to form a gelatin, the resulting bone gel is formed into sheets having a thickness (t). 2. The bone gel composition of claim 1 wherein the composition is formed into a polygonal shape. 3. The bone gel composition of claim 2 wherein the polygonal shape is a square or rectangle. 4. The bone gel composition of claim 2 wherein the polygonal shape is a circle. 5. The bone gel composition of claim 2 wherein the polygonal shape is any other multi-sided shape. 6. The bone gel composition of claim 1 wherein the thickness (t) is in the range of 1 to 10 mm. 7. The bone gel composition of claim 6 wherein the thickness (t) is in the range of 2 to 3 mm. 8. The bone gel composition of claim 3 wherein the rectangle has a predetermined width and a predetermined length. 9. The bone gel composition of claim 1 wherein the cortical bone has the cut pieces having a width, a length and a thickness in the range of 1 to 4 mm. 10. The bone gel composition of claim 9 wherein the cortical bone pieces are ground to a particle size up to 125 microns. 11. The bone gel composition of claim 1 wherein the bone gel formed into sheets is then frozen. 12. The bone gel composition of claim 11 wherein the bone gel sheet is frozen to −20 to −80 degrees C. 13. A method of making a bone gel composition consisting of cortical bone comprises the steps of:
preparing cortical bone by cutting the cortical bone into pieces, freeze-drying the pieces and then grinding into particles and demineralizing the ground particles and the freeze-drying the demineralized ground particles to form DBM particles; autoclaving a volume of the DBM particles mixed with sterile water in a 2:1 ratio by volume for a predetermined time at a pre-set temperature and pressure to form a gelatin; cooling the gelatin to form a bone gel; forming the bone gel into sheets of bone gel of a thickness (t); and freezing or packaging the sheets of bone gel, wherein the sheets of bone gel are frozen either before or after packaging. 14. The method of making a bone gel composition of claim 13 includes the step of cutting the sheets of bone gel to a desired circular, semi-circular or polygonal shape. 15. The method of making a bone gel composition of claim 14 wherein the polygonal shape is a square or rectangle. 16. The method of making a bone gel composition of claim 15 wherein the polygonal shape is a rectangle. 17. The method of making a bone gel composition of claim 13 wherein the thickness (t) is in the range of 1 to 10 mm. 18. The method of making a bone gel composition of claim 17 wherein the thickness (t) is in the range of 2 to 3 mm. 19. The method of making a bone gel composition of claim 13 wherein the rectangle has a predetermined width and a predetermined length. 20. The method of making a bone gel composition of claim 13 wherein the bone gel sheet is frozen at −20 to −80 degrees C. 21. The method of making a bone gel composition of claim 13 wherein the cortical bone has the cut pieces having a width, a length and a thickness in the range of 1 to 4 mm. 22. The method of making a bone gel composition of claim 13 wherein the cortical bone pieces are ground to a particle size up to 125 microns. | A bone gel composition consists of cortical bone. The cortical bone is made from cut pieces freeze-dried then ground into particles and demineralized then freeze-dried. A volume of the particles is placed in a solution of sterile water to create a mixture, the water volume being at least twice the particle volume, the mixture is autoclaved under heat and pressure to form a gelatin, the resulting bone gel is formed into sheets having a thickness (t).1. A bone gel composition consisting of cortical bone consists of cortical bone made from cut pieces freeze dried then ground into particles and demineralized then freeze-dried, wherein a volume of the particles is placed in a solution of sterile water to create a mixture, the water volume being at least twice the volume of the particles, the mixture is autoclaved under heat and pressure to form a gelatin, the resulting bone gel is formed into sheets having a thickness (t). 2. The bone gel composition of claim 1 wherein the composition is formed into a polygonal shape. 3. The bone gel composition of claim 2 wherein the polygonal shape is a square or rectangle. 4. The bone gel composition of claim 2 wherein the polygonal shape is a circle. 5. The bone gel composition of claim 2 wherein the polygonal shape is any other multi-sided shape. 6. The bone gel composition of claim 1 wherein the thickness (t) is in the range of 1 to 10 mm. 7. The bone gel composition of claim 6 wherein the thickness (t) is in the range of 2 to 3 mm. 8. The bone gel composition of claim 3 wherein the rectangle has a predetermined width and a predetermined length. 9. The bone gel composition of claim 1 wherein the cortical bone has the cut pieces having a width, a length and a thickness in the range of 1 to 4 mm. 10. The bone gel composition of claim 9 wherein the cortical bone pieces are ground to a particle size up to 125 microns. 11. The bone gel composition of claim 1 wherein the bone gel formed into sheets is then frozen. 12. The bone gel composition of claim 11 wherein the bone gel sheet is frozen to −20 to −80 degrees C. 13. A method of making a bone gel composition consisting of cortical bone comprises the steps of:
preparing cortical bone by cutting the cortical bone into pieces, freeze-drying the pieces and then grinding into particles and demineralizing the ground particles and the freeze-drying the demineralized ground particles to form DBM particles; autoclaving a volume of the DBM particles mixed with sterile water in a 2:1 ratio by volume for a predetermined time at a pre-set temperature and pressure to form a gelatin; cooling the gelatin to form a bone gel; forming the bone gel into sheets of bone gel of a thickness (t); and freezing or packaging the sheets of bone gel, wherein the sheets of bone gel are frozen either before or after packaging. 14. The method of making a bone gel composition of claim 13 includes the step of cutting the sheets of bone gel to a desired circular, semi-circular or polygonal shape. 15. The method of making a bone gel composition of claim 14 wherein the polygonal shape is a square or rectangle. 16. The method of making a bone gel composition of claim 15 wherein the polygonal shape is a rectangle. 17. The method of making a bone gel composition of claim 13 wherein the thickness (t) is in the range of 1 to 10 mm. 18. The method of making a bone gel composition of claim 17 wherein the thickness (t) is in the range of 2 to 3 mm. 19. The method of making a bone gel composition of claim 13 wherein the rectangle has a predetermined width and a predetermined length. 20. The method of making a bone gel composition of claim 13 wherein the bone gel sheet is frozen at −20 to −80 degrees C. 21. The method of making a bone gel composition of claim 13 wherein the cortical bone has the cut pieces having a width, a length and a thickness in the range of 1 to 4 mm. 22. The method of making a bone gel composition of claim 13 wherein the cortical bone pieces are ground to a particle size up to 125 microns. | 1,600 |
1,071 | 16,687,447 | 1,623 | Administration of low molecular weight (10,000-20,000 Daltons, or lower) pectins, particularly modified citrus pectins (MCP), like PectaSol-C reduces galectins-3 levels in vivo. Reduction of galectin-3 levels by MCP inhibits inflammation, inhibits fibrosis formation in organs and tissues, and inhibits cancer formation, progression, transformation and metastases. The reduction in circulating, serum and cellular galectin-3, inherently resulting from the administration of MCP, provides benefit over a spectrum of biological conditions, as evidenced by in vivo trials. | 1-6. (canceled) 7. A method of treating a mammal which benefits from a reduction in available circulating galectin-3, comprising the steps of:
a) Identifying a mammal in need of treatment for kidney damage, and b) Administering to said mammal an amount of modified pectin of molecular weight of 3,000-13,000 Daltons, in an amount of 10-750 mg/kg/day, for a period of time sufficient such that said mammal exhibits a reduction in active galectin-3 levels in said mammal and thereby treat kidney damage in said mammal. 8. A method of treating a mammal which benefits from a reduction in available galectin-3, comprising the steps of:
a) Identifying a mammal in need of treatment for kidney damage, and b) Administering to said mammal modified pectin of low molecular weight of 10,000-20,000 Daltons, in an amount of 5-1,500 mg/kg/day, for a period of time sufficient for said mammal to exhibit a reduction in active galectin-3 levels in said mammal and thereby treat kidney damage in said mammal. | Administration of low molecular weight (10,000-20,000 Daltons, or lower) pectins, particularly modified citrus pectins (MCP), like PectaSol-C reduces galectins-3 levels in vivo. Reduction of galectin-3 levels by MCP inhibits inflammation, inhibits fibrosis formation in organs and tissues, and inhibits cancer formation, progression, transformation and metastases. The reduction in circulating, serum and cellular galectin-3, inherently resulting from the administration of MCP, provides benefit over a spectrum of biological conditions, as evidenced by in vivo trials.1-6. (canceled) 7. A method of treating a mammal which benefits from a reduction in available circulating galectin-3, comprising the steps of:
a) Identifying a mammal in need of treatment for kidney damage, and b) Administering to said mammal an amount of modified pectin of molecular weight of 3,000-13,000 Daltons, in an amount of 10-750 mg/kg/day, for a period of time sufficient such that said mammal exhibits a reduction in active galectin-3 levels in said mammal and thereby treat kidney damage in said mammal. 8. A method of treating a mammal which benefits from a reduction in available galectin-3, comprising the steps of:
a) Identifying a mammal in need of treatment for kidney damage, and b) Administering to said mammal modified pectin of low molecular weight of 10,000-20,000 Daltons, in an amount of 5-1,500 mg/kg/day, for a period of time sufficient for said mammal to exhibit a reduction in active galectin-3 levels in said mammal and thereby treat kidney damage in said mammal. | 1,600 |
1,072 | 13,824,548 | 1,619 | The present invention relates to a cosmetic composition comprising, in a physiologically acceptable medium, at least water, at least alkylcellulose, at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils, and at least a second hydrocarbon-based nonvolatile oil chosen from C10-C26 alcohols, preferably monoalcohols; monoesters, dieters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol, which are optionally hydroxylated; and polyesters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids. | 1-18. (canceled) 19. A cosmetic composition comprising, in a physiologically acceptable medium:
at least 5% by weight of water; at least alkylcellulose, the alkyl residue of which comprises between 1 and 6 carbon atoms; at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils; and at least a second hydrocarbon-based nonvolatile oil, chosen from: C10-C26 alcohols; optionally hydroxylated monoesters, diesters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol; esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids. 20. The composition as claimed in claim 19, wherein the alkylcellulose is present in a content of between 1% and 60% by weight, relative to the total weight of the composition. 21. The composition as claimed in claim 19, wherein the alkylcellulose is present in a content of between 5% and 30% by weight, relative to the total weight of the composition. 22. The composition as claimed in claim 19, wherein the alkylcellulose is present in a content of between 5% and 20% by weight, relative to the total weight of the composition. 23. The composition as claimed in claim 19, wherein the alkylcellulose is chosen from methylcellulose, ethylcellulose and propylcellulose. 24. The composition as claimed in claim 19, wherein the alkylcellulose is ethylcellulose. 25. The composition as claimed in claim 19, wherein said first nonvolatile oil is chosen from phenyl silicone oils and fluoro oils. 26. The composition as claimed in claim 19, said composition comprising a content ranging from 5% to 75% by weight of first nonvolatile silicone oil(s) and/or fluoro oil(s)t, relative to its total weight. 27. The composition as claimed in claim 19, said composition comprising a content ranging from 10% to 40% by weight, relative to its total weight. 28. The composition as claimed in claim 19, said composition comprising a content ranging from 15% to 30% by weight, relative to its total weight. 29. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based “second oil” is chosen from:
C10-C26 monoalcohols, such as lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, palmityl alcohol, oleyl alcohol, cetearyl alcohol (mixture of cetyl alcohol and stearyl alcohol), behenyl alcohol, erucyl alcohol, arachidyl alcohol, 2-hexyldecyl alcohol, isocetyl alcohol and octyldodecanol, and mixtures thereof;
optionally hydroxylated monoesters of a C2-C8 carboxylic acid and of a C2-C8 alcohol;
optionally hydroxylated diesters of a C2-C8 dicarboxylic acid and of a C2-C8 alcohol, such as diisopropyl adipate, 2-diethylhexyl adipate, dibutyl adipate or diisostearyl adipate;
optionally hydroxylated triesters of a C2-C8 tricarboxylic acid and of a C2-C8 alcohol, such as citric acid esters, such as trioctyl citrate, triethyl citrate, acetyl tributyl citrate, tributyl citrate or acetyl tributyl citrate,
esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids, such as glycol diesters of monoacids, such as neopentyl glycol diheptanoate, or glycol triesters of monoacids, such as triacetin. 30. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil(s) are present in a total content ranging from 5% to 75% by weight, relative to the total weight of the composition. 31. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil(s) are present in a total content ranging from 10% to 50% by weight, relative to the total weight of the composition. 32. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil(s) are present in a total content ranging from 20% to 45% by weight, relative to the total weight of the composition. 33. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil and the alkylcellulose are used in the composition according to the invention in a nonvolatile hydrocarbon-based second oil(s)/alkylcellulose weight ratio of between 1 and 20. 34. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil and the alkylcellulose are used in the composition according to the invention in a nonvolatile hydrocarbon-based second oil(s)/alkylcellulose weight ratio of between 2 and 15. 35. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil and the alkylcellulose are used in the composition according to the invention in a nonvolatile hydrocarbon-based second oil(s)/alkylcellulose weight ratio of between 3 and 10. 36. The composition as claimed in claim 19, said composition comprising between 5% and 80% by weight of water, relative to the total weight of the composition. 37. The composition as claimed in claim 19, said composition comprising between 15% and 50% by weight of water, relative to the total weight of the composition. 38. The composition as claimed in claim 19, said composition comprising:
between 4% and 30% by weight of alkylcellulose, between 15% and 50% by weight of water, between 45% and 75% by weight of nonvolatile oils. 39. The composition as claimed in claim 19, said composition comprising at least one dyestuff. 40. The composition as claimed in claim 19, said composition comprising at least one compound chosen from fillers, waxes, pasty fatty substances, semicrystalline polymers and/or lipophilic gelling agents, silicone gums, organopolysiloxane elastomers and silicone resins, and mixtures thereof. 41. The composition as claimed in claim 19, said composition being in liquid form. 42. The composition as claimed in claim 19, said composition being in the form of an oil-in-water emulsion. 43. The composition as claimed in claim 19, said composition being a composition for making up and/or caring for the lips or the skin. 44. The composition as claimed in claim 19, said composition being a lipstick. 45. A cosmetic process for making up and/or caring for the skin and/or the lips, comprising at least one step that consists in applying to the skin and/or the lips at least one composition comprising, in a physiologically acceptable medium:
at least 5% by weight of water; at least alkylcellulose, the alkyl residue of which comprises between 1 and 6 carbon atoms; at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils; and at least a second hydrocarbon-based nonvolatile oil, chosen from: C10-C26 alcohols; optionally hydroxylated monoesters, diesters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol; esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids. 46. A cosmetic process for making up and/or caring for the lips, comprising at least one step that consists in applying to the lips at least one cosmetic composition comprising, in a physiologically acceptable medium:
at least water; at least alkylcellulose, the alkyl residue of which comprises between 1 and 6 carbon atoms; at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils; and at least a second hydrocarbon-based nonvolatile oil, chosen from: C10-C26 alcohols; optionally hydroxylated monoesters, diesters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol; esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids. 47. A process for preparing a composition comprising, in a physiologically acceptable medium:
at least 5% by weight of water; at least alkylcellulose, the alkyl residue of which comprises between 1 and 6 carbon atoms; at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils; and at least a second hydrocarbon-based nonvolatile oil, chosen from: C10-C26 alcohols; optionally hydroxylated monoesters, diesters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol; esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids, wherein the alkylcellulose is used therein in the form of a stable aqueous dispersion. | The present invention relates to a cosmetic composition comprising, in a physiologically acceptable medium, at least water, at least alkylcellulose, at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils, and at least a second hydrocarbon-based nonvolatile oil chosen from C10-C26 alcohols, preferably monoalcohols; monoesters, dieters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol, which are optionally hydroxylated; and polyesters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids.1-18. (canceled) 19. A cosmetic composition comprising, in a physiologically acceptable medium:
at least 5% by weight of water; at least alkylcellulose, the alkyl residue of which comprises between 1 and 6 carbon atoms; at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils; and at least a second hydrocarbon-based nonvolatile oil, chosen from: C10-C26 alcohols; optionally hydroxylated monoesters, diesters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol; esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids. 20. The composition as claimed in claim 19, wherein the alkylcellulose is present in a content of between 1% and 60% by weight, relative to the total weight of the composition. 21. The composition as claimed in claim 19, wherein the alkylcellulose is present in a content of between 5% and 30% by weight, relative to the total weight of the composition. 22. The composition as claimed in claim 19, wherein the alkylcellulose is present in a content of between 5% and 20% by weight, relative to the total weight of the composition. 23. The composition as claimed in claim 19, wherein the alkylcellulose is chosen from methylcellulose, ethylcellulose and propylcellulose. 24. The composition as claimed in claim 19, wherein the alkylcellulose is ethylcellulose. 25. The composition as claimed in claim 19, wherein said first nonvolatile oil is chosen from phenyl silicone oils and fluoro oils. 26. The composition as claimed in claim 19, said composition comprising a content ranging from 5% to 75% by weight of first nonvolatile silicone oil(s) and/or fluoro oil(s)t, relative to its total weight. 27. The composition as claimed in claim 19, said composition comprising a content ranging from 10% to 40% by weight, relative to its total weight. 28. The composition as claimed in claim 19, said composition comprising a content ranging from 15% to 30% by weight, relative to its total weight. 29. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based “second oil” is chosen from:
C10-C26 monoalcohols, such as lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, palmityl alcohol, oleyl alcohol, cetearyl alcohol (mixture of cetyl alcohol and stearyl alcohol), behenyl alcohol, erucyl alcohol, arachidyl alcohol, 2-hexyldecyl alcohol, isocetyl alcohol and octyldodecanol, and mixtures thereof;
optionally hydroxylated monoesters of a C2-C8 carboxylic acid and of a C2-C8 alcohol;
optionally hydroxylated diesters of a C2-C8 dicarboxylic acid and of a C2-C8 alcohol, such as diisopropyl adipate, 2-diethylhexyl adipate, dibutyl adipate or diisostearyl adipate;
optionally hydroxylated triesters of a C2-C8 tricarboxylic acid and of a C2-C8 alcohol, such as citric acid esters, such as trioctyl citrate, triethyl citrate, acetyl tributyl citrate, tributyl citrate or acetyl tributyl citrate,
esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids, such as glycol diesters of monoacids, such as neopentyl glycol diheptanoate, or glycol triesters of monoacids, such as triacetin. 30. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil(s) are present in a total content ranging from 5% to 75% by weight, relative to the total weight of the composition. 31. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil(s) are present in a total content ranging from 10% to 50% by weight, relative to the total weight of the composition. 32. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil(s) are present in a total content ranging from 20% to 45% by weight, relative to the total weight of the composition. 33. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil and the alkylcellulose are used in the composition according to the invention in a nonvolatile hydrocarbon-based second oil(s)/alkylcellulose weight ratio of between 1 and 20. 34. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil and the alkylcellulose are used in the composition according to the invention in a nonvolatile hydrocarbon-based second oil(s)/alkylcellulose weight ratio of between 2 and 15. 35. The composition as claimed in claim 19, wherein said nonvolatile hydrocarbon-based second oil and the alkylcellulose are used in the composition according to the invention in a nonvolatile hydrocarbon-based second oil(s)/alkylcellulose weight ratio of between 3 and 10. 36. The composition as claimed in claim 19, said composition comprising between 5% and 80% by weight of water, relative to the total weight of the composition. 37. The composition as claimed in claim 19, said composition comprising between 15% and 50% by weight of water, relative to the total weight of the composition. 38. The composition as claimed in claim 19, said composition comprising:
between 4% and 30% by weight of alkylcellulose, between 15% and 50% by weight of water, between 45% and 75% by weight of nonvolatile oils. 39. The composition as claimed in claim 19, said composition comprising at least one dyestuff. 40. The composition as claimed in claim 19, said composition comprising at least one compound chosen from fillers, waxes, pasty fatty substances, semicrystalline polymers and/or lipophilic gelling agents, silicone gums, organopolysiloxane elastomers and silicone resins, and mixtures thereof. 41. The composition as claimed in claim 19, said composition being in liquid form. 42. The composition as claimed in claim 19, said composition being in the form of an oil-in-water emulsion. 43. The composition as claimed in claim 19, said composition being a composition for making up and/or caring for the lips or the skin. 44. The composition as claimed in claim 19, said composition being a lipstick. 45. A cosmetic process for making up and/or caring for the skin and/or the lips, comprising at least one step that consists in applying to the skin and/or the lips at least one composition comprising, in a physiologically acceptable medium:
at least 5% by weight of water; at least alkylcellulose, the alkyl residue of which comprises between 1 and 6 carbon atoms; at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils; and at least a second hydrocarbon-based nonvolatile oil, chosen from: C10-C26 alcohols; optionally hydroxylated monoesters, diesters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol; esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids. 46. A cosmetic process for making up and/or caring for the lips, comprising at least one step that consists in applying to the lips at least one cosmetic composition comprising, in a physiologically acceptable medium:
at least water; at least alkylcellulose, the alkyl residue of which comprises between 1 and 6 carbon atoms; at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils; and at least a second hydrocarbon-based nonvolatile oil, chosen from: C10-C26 alcohols; optionally hydroxylated monoesters, diesters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol; esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids. 47. A process for preparing a composition comprising, in a physiologically acceptable medium:
at least 5% by weight of water; at least alkylcellulose, the alkyl residue of which comprises between 1 and 6 carbon atoms; at least a first nonvolatile oil chosen from silicone oils and/or fluoro oils; and at least a second hydrocarbon-based nonvolatile oil, chosen from: C10-C26 alcohols; optionally hydroxylated monoesters, diesters or triesters of a C2-C8 monocarboxylic or polycarboxylic acid and of a C2-C8 alcohol; esters of a C2-C8 polyol and of one or more C2-C8 carboxylic acids, wherein the alkylcellulose is used therein in the form of a stable aqueous dispersion. | 1,600 |
1,073 | 15,706,177 | 1,655 | A composition useful for promoting a desired oral microbiota to treat an allergy related respiratory condition in a subject in need of such treatment, the composition including an amino acid containing ingredient including individual molecules of L-arginine; the composition in a configuration to be maintained within the oral cavity for a period of at least about 30 seconds to about an hour, the desired oral microbiota including Veillonella and Streptococcus. | 1. A composition useful for promoting a desired oral microbiota to treat an allergy related respiratory condition in a subject in need of such treatment comprising:
an amino acid containing ingredient comprising individual molecules of L-arginine; the composition in a configuration to be maintained within the oral cavity for a period of at least about 30 seconds to about an hour, the desired oral microbiota comprising Veillonella and Streptococcus. 2. The composition of claim 1, wherein the composition is comprises a carrier. 3. The composition of claim 2, wherein the carrier comprises at least one of a solid, powder, and liquid. 4. The composition of claim 2, wherein the carrier comprises one or more of a chewable tablet, an edible capsule, and a hygienic paste, and an edible food. 5. The composition of claim 1, wherein the at least one amino acid further comprises at least one of L-cysteine, DL-aspartic acid, L-glutamic acid, L-serine, and L-tyrosine. 6. The composition of claim 1, wherein the amino acid containing ingredient is present in the composition at a weight percent level of from about 0.1 wt. % to about 99.9 wt. %. 7. The composition of claim 1, wherein the amino acid containing ingredient is present in the composition at a weight percent level of from about 0.1 wt. % to about 95 wt. %. 8. The composition of claim 1, wherein the amino acid containing ingredient is present in the composition at a weight percent level of from about 5.0 wt. % to about 95.0 wt. %. 9. The composition of claim 1, wherein the composition further comprises a sugar containing ingredient, the sugar comprising one or more of monosaccharides, disaccharides, oligosaccharides, and polysaccharides. 10. The composition of claim 9, wherein the sugar comprises one or more of mannose, lactose, malibiose, maltose, cellobiose, trehalose, and raffinose. 11. The composition of claim 9, wherein the composition further comprises polysaccharide polymers. 12. The composition of claim 11, wherein the polysaccharide polymers comprise one or more of pullulan and inulin. 13. The composition of claim 9, wherein the sugar comprises one or more of rhamnose, xylitol, arabitol, fructose, glucose, inositol, sucrose, and maltose. 14. The composition of claim 1, wherein the composition further comprises one or more of carbohydrates, salts, flavorants, proteins, surfactants, emulsifiers, flavonoids, alcohols, synthetic sweeteners, and food preserving agents. 15. The composition of claim 1, wherein the composition further comprises one or more of brown sugar, syrup, honey, chocolate, nuts, almonds, spices, cinnamon, and vanilla. 16. The composition of claim 9, wherein the sugar is present in the composition at a weight percent level of from about 0.1 wt. % to about 95 wt. %. 17. The composition of claim 9, wherein the sugar is present in the composition at a weight percent level of from about 0.5 wt. % to about 95 wt. %. 18. The composition of claim 9, wherein the sugar is present in the composition at a weight percent level of from about 5.0 wt. % to about 95.0 wt. %. 19. The composition of claim 1, further comprising live bacteria comprising a lactic acid producing bacterium and a lactate fermenting bacterium at a level of from about 1000 to about 1,000,000,000 living cells. 20. The composition of claim 1 wherein the Veillonella comprises one or more of Veillonella Dispar and Veillonella Parvula and wherein the Streptococcus comprises one or more of Streptococcus salivarius and Streptococcus thermophilus. | A composition useful for promoting a desired oral microbiota to treat an allergy related respiratory condition in a subject in need of such treatment, the composition including an amino acid containing ingredient including individual molecules of L-arginine; the composition in a configuration to be maintained within the oral cavity for a period of at least about 30 seconds to about an hour, the desired oral microbiota including Veillonella and Streptococcus.1. A composition useful for promoting a desired oral microbiota to treat an allergy related respiratory condition in a subject in need of such treatment comprising:
an amino acid containing ingredient comprising individual molecules of L-arginine; the composition in a configuration to be maintained within the oral cavity for a period of at least about 30 seconds to about an hour, the desired oral microbiota comprising Veillonella and Streptococcus. 2. The composition of claim 1, wherein the composition is comprises a carrier. 3. The composition of claim 2, wherein the carrier comprises at least one of a solid, powder, and liquid. 4. The composition of claim 2, wherein the carrier comprises one or more of a chewable tablet, an edible capsule, and a hygienic paste, and an edible food. 5. The composition of claim 1, wherein the at least one amino acid further comprises at least one of L-cysteine, DL-aspartic acid, L-glutamic acid, L-serine, and L-tyrosine. 6. The composition of claim 1, wherein the amino acid containing ingredient is present in the composition at a weight percent level of from about 0.1 wt. % to about 99.9 wt. %. 7. The composition of claim 1, wherein the amino acid containing ingredient is present in the composition at a weight percent level of from about 0.1 wt. % to about 95 wt. %. 8. The composition of claim 1, wherein the amino acid containing ingredient is present in the composition at a weight percent level of from about 5.0 wt. % to about 95.0 wt. %. 9. The composition of claim 1, wherein the composition further comprises a sugar containing ingredient, the sugar comprising one or more of monosaccharides, disaccharides, oligosaccharides, and polysaccharides. 10. The composition of claim 9, wherein the sugar comprises one or more of mannose, lactose, malibiose, maltose, cellobiose, trehalose, and raffinose. 11. The composition of claim 9, wherein the composition further comprises polysaccharide polymers. 12. The composition of claim 11, wherein the polysaccharide polymers comprise one or more of pullulan and inulin. 13. The composition of claim 9, wherein the sugar comprises one or more of rhamnose, xylitol, arabitol, fructose, glucose, inositol, sucrose, and maltose. 14. The composition of claim 1, wherein the composition further comprises one or more of carbohydrates, salts, flavorants, proteins, surfactants, emulsifiers, flavonoids, alcohols, synthetic sweeteners, and food preserving agents. 15. The composition of claim 1, wherein the composition further comprises one or more of brown sugar, syrup, honey, chocolate, nuts, almonds, spices, cinnamon, and vanilla. 16. The composition of claim 9, wherein the sugar is present in the composition at a weight percent level of from about 0.1 wt. % to about 95 wt. %. 17. The composition of claim 9, wherein the sugar is present in the composition at a weight percent level of from about 0.5 wt. % to about 95 wt. %. 18. The composition of claim 9, wherein the sugar is present in the composition at a weight percent level of from about 5.0 wt. % to about 95.0 wt. %. 19. The composition of claim 1, further comprising live bacteria comprising a lactic acid producing bacterium and a lactate fermenting bacterium at a level of from about 1000 to about 1,000,000,000 living cells. 20. The composition of claim 1 wherein the Veillonella comprises one or more of Veillonella Dispar and Veillonella Parvula and wherein the Streptococcus comprises one or more of Streptococcus salivarius and Streptococcus thermophilus. | 1,600 |
1,074 | 15,630,465 | 1,612 | This invention relates to oral care compositions comprising a first source of zinc which is zinc citrate, a second source of zinc which is zinc oxide or zinc lactate, a stannous ion source, and a polyphosphate, as well as to methods of using and of making these compositions. | 1. An oral care composition comprising:
a. A first source of zinc comprising zinc oxide; b. A second source of zinc comprising zinc citrate or zinc lactate; and c. stannous ion source. 2. The oral care composition of claim 1, wherein the first zinc source is zinc oxide and the second zinc source is zinc citrate. 3. The oral care composition of claim 1, wherein the first zinc source is zinc oxide and the second source is zinc lactate. 4. The oral composition of claim 1, further comprising a polyphosphate. 5. The oral care composition of claim 1, comprising zinc citrate and zinc oxide, wherein the zinc citrate is in an amount of from 0.25 to 1 wt. % and zinc oxide is present in an amount of from 0.75 to 1.25 wt. % based on the total weight of the composition. 6. The oral care composition of claim 1, comprising zinc lactate and zinc oxide, wherein the zinc lactate is in an amount of from 0.5 to 0.9 wt. % and zinc oxide is present in an amount of from 0.75 to 1.25 wt. %, based on the total weight of the composition. 7. The oral care composition of claim 1, wherein the stannous ion source is stannous fluoride, other stannous halides such as stannous chloride dihydrate, stannous pyrophosphate, organic stannous carboxylate salts such as stannous formate, acetate, gluconate, lactate, tartrate, oxalate, malonate and citrate, stannous ethylene glyoxide, or a mixture thereof. 8. The oral care composition of claim 7, wherein the stannous source is stannous fluoride. 9. The oral care composition of claim 8, wherein the stannous fluoride is present in an amount of 0.1 wt. % to 2 wt. % based on the total weight of the composition. 10. The oral care composition of claim 8, wherein the stannous fluoride is in an amount from 50 to 25,000 ppm. 11. The oral care composition of claim 1, wherein composition comprises a copolymer. 12. The oral care composition of claim 11, wherein the copolymer is a PVM/MA copolymer. 13. The oral care composition of claim 1, wherein the polyphosphate is sodium tripolyphosphate 14. The oral care composition of claim 13, wherein the sodium tripolyphosphate is present in an amount of from 0.5-5.0 wt %, based on the total weight of the composition. 15. The oral care composition of claim 1, wherein the composition further comprises glycerin, wherein the glycerin is in a total amount of 20-500%, based on the total weight of the composition. 16. The oral care composition of claim 1, wherein the ratio of the amount of zinc oxide (wt. %) to zinc citrate (wt %) is from 1.5:1 to 4.5:1. 17. The oral care composition of claim 1 further comprising a whitening agent. 18. The oral care composition of claim 1 comprising:
a. about 1.0% zinc oxide
b. about 0.5% zinc citrate
c. about 4500 ppm stannous fluoride;
d. about 3.0% of sodium tripolyphosphate 19. The oral care composition of claim 1 comprising:
e. about 1.0% zinc oxide
f. about 0.5% zinc citrate
g. about 4500 ppm stannous fluoride;
h. about 3.0% of sodium tripolyphosphate; and
i. about 41% glycerin 20. The oral care composition of claim 1, wherein the oral composition may be any of the following oral compositions selected from the group consisting of: a toothpaste or a dentifrice, a mouthwash or a mouth rinse, a topical oral gel, a chewing gum, and a denture cleanser. 21. The oral care composition of claim 1, wherein the composition is obtained or obtainable by combining the ingredients as set forth in any of the preceding compositions. 22. A method to improve oral health comprising applying an effective amount of the oral composition of claim 1 to the oral cavity of a subject in need thereof. | This invention relates to oral care compositions comprising a first source of zinc which is zinc citrate, a second source of zinc which is zinc oxide or zinc lactate, a stannous ion source, and a polyphosphate, as well as to methods of using and of making these compositions.1. An oral care composition comprising:
a. A first source of zinc comprising zinc oxide; b. A second source of zinc comprising zinc citrate or zinc lactate; and c. stannous ion source. 2. The oral care composition of claim 1, wherein the first zinc source is zinc oxide and the second zinc source is zinc citrate. 3. The oral care composition of claim 1, wherein the first zinc source is zinc oxide and the second source is zinc lactate. 4. The oral composition of claim 1, further comprising a polyphosphate. 5. The oral care composition of claim 1, comprising zinc citrate and zinc oxide, wherein the zinc citrate is in an amount of from 0.25 to 1 wt. % and zinc oxide is present in an amount of from 0.75 to 1.25 wt. % based on the total weight of the composition. 6. The oral care composition of claim 1, comprising zinc lactate and zinc oxide, wherein the zinc lactate is in an amount of from 0.5 to 0.9 wt. % and zinc oxide is present in an amount of from 0.75 to 1.25 wt. %, based on the total weight of the composition. 7. The oral care composition of claim 1, wherein the stannous ion source is stannous fluoride, other stannous halides such as stannous chloride dihydrate, stannous pyrophosphate, organic stannous carboxylate salts such as stannous formate, acetate, gluconate, lactate, tartrate, oxalate, malonate and citrate, stannous ethylene glyoxide, or a mixture thereof. 8. The oral care composition of claim 7, wherein the stannous source is stannous fluoride. 9. The oral care composition of claim 8, wherein the stannous fluoride is present in an amount of 0.1 wt. % to 2 wt. % based on the total weight of the composition. 10. The oral care composition of claim 8, wherein the stannous fluoride is in an amount from 50 to 25,000 ppm. 11. The oral care composition of claim 1, wherein composition comprises a copolymer. 12. The oral care composition of claim 11, wherein the copolymer is a PVM/MA copolymer. 13. The oral care composition of claim 1, wherein the polyphosphate is sodium tripolyphosphate 14. The oral care composition of claim 13, wherein the sodium tripolyphosphate is present in an amount of from 0.5-5.0 wt %, based on the total weight of the composition. 15. The oral care composition of claim 1, wherein the composition further comprises glycerin, wherein the glycerin is in a total amount of 20-500%, based on the total weight of the composition. 16. The oral care composition of claim 1, wherein the ratio of the amount of zinc oxide (wt. %) to zinc citrate (wt %) is from 1.5:1 to 4.5:1. 17. The oral care composition of claim 1 further comprising a whitening agent. 18. The oral care composition of claim 1 comprising:
a. about 1.0% zinc oxide
b. about 0.5% zinc citrate
c. about 4500 ppm stannous fluoride;
d. about 3.0% of sodium tripolyphosphate 19. The oral care composition of claim 1 comprising:
e. about 1.0% zinc oxide
f. about 0.5% zinc citrate
g. about 4500 ppm stannous fluoride;
h. about 3.0% of sodium tripolyphosphate; and
i. about 41% glycerin 20. The oral care composition of claim 1, wherein the oral composition may be any of the following oral compositions selected from the group consisting of: a toothpaste or a dentifrice, a mouthwash or a mouth rinse, a topical oral gel, a chewing gum, and a denture cleanser. 21. The oral care composition of claim 1, wherein the composition is obtained or obtainable by combining the ingredients as set forth in any of the preceding compositions. 22. A method to improve oral health comprising applying an effective amount of the oral composition of claim 1 to the oral cavity of a subject in need thereof. | 1,600 |
1,075 | 15,520,795 | 1,651 | A tissue sample processing system and associated methods is disclosed and described. The tissue sample processing system ( 100 ) can include a microfluidic separating system ( 110 ). The microfluidic separating system ( 110 ) can include a fluid channel to receive a carrier fluid ( 104 ) and a tissue sample ( 102 ), and a plurality of outlets. Flow of the carrier fluid ( 104 ) and the tissue sample ( 102 ) in the fluid channel can facilitate segregation of materials in the tissue sample ( 102 ) based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the tissue sample. In addition, the sample processing system ( 100 ) can comprise a cryopreservation system ( 120 ) associated with at least one of the plurality of outlets to freeze the material in the tissue sample ( 102 ) associated with the at least one of the plurality of outlets. | 1. A method of separating sperm cells, comprising:
obtaining a microfluidic separating system having a fluid channel and a plurality of outlets; and disposing a sperm sample in the fluid channel, wherein flow of the sperm sample in the fluid channel facilitates segregation of materials in the sperm sample based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the sperm sample. 2. The method of claim 1, wherein the fluid channel comprises a spiral configuration. 3. The method of claim 1, wherein the microfluidic separating system comprises:
an inlet zone having a carrier fluid inlet to receive a carrier fluid and a tissue sample inlet to receive the tissue sample; an outlet zone having the plurality of outlets; and a transport region between the inlet zone and the outlet zone, the transport region being open to the carrier fluid and the tissue sample, wherein a cross-flow in the transport region facilitates segregation of the materials in the tissue sample based on size. 4. The method of claim 1, wherein the material in the sperm sample associated with at least one of the plurality of outlets comprises non-motile sperm cells. 5. The method of claim 1, further comprising:
associating a sorting system with at least one of the plurality of outlets, wherein the material in the sperm sample associated with the at least one of the plurality of outlets comprises non-motile sperm cells; and sorting the non-motile sperm cells into a plurality of aliquots. 6. The method of claim 5, wherein the sorting system comprises:
an inlet to receive the material in the tissue sample associated with the at least one of the plurality of outlets; a plurality of aliquot storage channels; and a plurality of valves associated with the plurality of aliquot storage channels to facilitate separation of the plurality of aliquots into the aliquot storage channels. 7. A method of separating non-motile sperm cells from a sperm sample, comprising:
flowing a sperm sample through a fluid channel under laminar flow conditions, wherein a cross-flow in the fluid channel facilitates segregation of non-motile sperm cells within an inner fluid flow layer; and spatially separating the inner fluid flow layer. 8. The method of claim 7, wherein the fluid channel comprises a spiral configuration. 9. The method of claim 7, further comprising receiving the non-motile sperm cells in an outlet of the fluid channel. 10. The method of claim 9, further comprising sorting the non-motile sperm cells into a plurality of aliquots. 11. The method of claim 10, wherein each of the plurality of aliquots comprises from about 1 to about 20 non-motile sperm cells. 12. The method of claim 11, wherein each of the plurality of aliquots comprises from about 1 to about 10 non-motile sperm cells. 13. The method of claim 12, wherein each of the plurality of aliquots comprises a single non-motile sperm cell. 14. A tissue sample processing system, comprising:
a microfluidic separating system having
a fluid channel to receive a sample fluid including a carrier fluid and a tissue sample, and
a plurality of outlets, wherein flow of the carrier fluid and the tissue sample in the fluid channel facilitates segregation of materials in the tissue sample based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the tissue sample; and
a cryopreservation system associated with at least one of the plurality of outlets to freeze the material in the tissue sample associated with the at least one of the plurality of outlets. 15. The system of claim 14, wherein the fluid channel comprises a spiral configuration. 16. The system of claim 14, wherein the microfluidic separating system comprises:
an inlet zone having a carrier fluid inlet to receive the carrier fluid and a tissue sample inlet to receive the tissue sample; an outlet zone having the plurality of outlets; and a transport region between the inlet zone and the outlet zone, the transport region being open to the carrier fluid and the tissue sample, wherein a cross-flow in the transport region facilitates segregation of the materials in the tissue sample based on size. 17. The system of claim 14, wherein the tissue sample comprises a sperm sample. 18. The system of claim 14, wherein the material in the tissue sample associated with the at least one of the plurality of outlets comprises non-motile sperm cells. 19. A tissue sample processing system, comprising:
a microfluidic separating system having
a fluid channel to receive a carrier fluid and a tissue sample, and
a plurality of outlets, wherein flow of the carrier fluid and the tissue sample in the fluid channel facilitates segregation of materials in the tissue sample based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the tissue sample; and
a sorting system associated with at least one of the plurality of outlets to sort a plurality of aliquots of the material in the tissue sample associated with the at least one of the plurality of outlets. 20. The system of claim 19, wherein the sorting system comprises:
an inlet to receive the material in the tissue sample associated with the at least one of the plurality of outlets; a plurality of aliquot storage channels; and a plurality of valves associated with the plurality of aliquot storage channels to facilitate separation of the plurality of aliquots into the aliquot storage channels. 21. The system of claim 19, wherein the fluid channel comprises a spiral configuration. 22. The system of claim 19, wherein the microfluidic separating system comprises:
an inlet zone having a carrier fluid inlet to receive the carrier fluid and a tissue sample inlet to receive the tissue sample; an outlet zone having the plurality of outlets; and a transport region between the inlet zone and the outlet zone, the transport region being open to the carrier fluid and the tissue sample, wherein a cross-flow in the transport region facilitates segregation of the materials in the tissue sample based on size. 23. The system of claim 19, further comprising a cryopreservation system associated with the sorting system to freeze at least one of the plurality of aliquots of the material in the tissue sample. 24. The system of claim 19, wherein each of the plurality of aliquots comprises from about 1 to about 20 non-motile sperm cells. 25. The system of claim 24, wherein each of the plurality of aliquots comprises from about 1 to about 10 non-motile sperm cells. 26. The system of claim 25, wherein each of the plurality of aliquots comprises a single non-motile sperm cell. 27. A tissue sample processing system, comprising:
a microfluidic separating system having
a fluid channel to receive a carrier fluid and a tissue sample, and
a plurality of outlets, wherein flow of the carrier fluid and the tissue sample in the fluid channel facilitates segregation of materials in the tissue sample based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the tissue sample; and
a concentrating system associated with at least one of the plurality of outlets to concentrate a size fraction of the material in the tissue sample associated with the at least one of the plurality of outlets. 28. The system of claim 27, wherein the concentrating system comprises a first fluid conduit and a second fluid conduit, wherein the first fluid conduit is sized to prevent passage of the size fraction of the material therethrough, and wherein a fluid resistance of the first fluid conduit is greater than a fluid resistance of the second fluid conduit such that the size fraction of material flows towards the first conduit and is trapped at the first conduit. 29. The system of claim 27, wherein the fluid channel comprises a spiral configuration. 30. The system of claim 27, wherein the microfluidic separating system comprises:
an inlet zone having a carrier fluid inlet to receive the carrier fluid and a tissue sample inlet to receive the tissue sample; an outlet zone having the plurality of outlets; and a transport region between the inlet zone and the outlet zone, the transport region being open to the carrier fluid and the tissue sample, wherein a cross-flow in the transport region facilitates segregation of the materials in the tissue sample based on size. 31. The system of claim 27, further comprising a cryopreservation system associated with the concentrating system to freeze the concentrated size fraction of the material in the tissue sample. 32. The system of claim 27, wherein the tissue sample comprises a sperm sample. | A tissue sample processing system and associated methods is disclosed and described. The tissue sample processing system ( 100 ) can include a microfluidic separating system ( 110 ). The microfluidic separating system ( 110 ) can include a fluid channel to receive a carrier fluid ( 104 ) and a tissue sample ( 102 ), and a plurality of outlets. Flow of the carrier fluid ( 104 ) and the tissue sample ( 102 ) in the fluid channel can facilitate segregation of materials in the tissue sample ( 102 ) based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the tissue sample. In addition, the sample processing system ( 100 ) can comprise a cryopreservation system ( 120 ) associated with at least one of the plurality of outlets to freeze the material in the tissue sample ( 102 ) associated with the at least one of the plurality of outlets.1. A method of separating sperm cells, comprising:
obtaining a microfluidic separating system having a fluid channel and a plurality of outlets; and disposing a sperm sample in the fluid channel, wherein flow of the sperm sample in the fluid channel facilitates segregation of materials in the sperm sample based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the sperm sample. 2. The method of claim 1, wherein the fluid channel comprises a spiral configuration. 3. The method of claim 1, wherein the microfluidic separating system comprises:
an inlet zone having a carrier fluid inlet to receive a carrier fluid and a tissue sample inlet to receive the tissue sample; an outlet zone having the plurality of outlets; and a transport region between the inlet zone and the outlet zone, the transport region being open to the carrier fluid and the tissue sample, wherein a cross-flow in the transport region facilitates segregation of the materials in the tissue sample based on size. 4. The method of claim 1, wherein the material in the sperm sample associated with at least one of the plurality of outlets comprises non-motile sperm cells. 5. The method of claim 1, further comprising:
associating a sorting system with at least one of the plurality of outlets, wherein the material in the sperm sample associated with the at least one of the plurality of outlets comprises non-motile sperm cells; and sorting the non-motile sperm cells into a plurality of aliquots. 6. The method of claim 5, wherein the sorting system comprises:
an inlet to receive the material in the tissue sample associated with the at least one of the plurality of outlets; a plurality of aliquot storage channels; and a plurality of valves associated with the plurality of aliquot storage channels to facilitate separation of the plurality of aliquots into the aliquot storage channels. 7. A method of separating non-motile sperm cells from a sperm sample, comprising:
flowing a sperm sample through a fluid channel under laminar flow conditions, wherein a cross-flow in the fluid channel facilitates segregation of non-motile sperm cells within an inner fluid flow layer; and spatially separating the inner fluid flow layer. 8. The method of claim 7, wherein the fluid channel comprises a spiral configuration. 9. The method of claim 7, further comprising receiving the non-motile sperm cells in an outlet of the fluid channel. 10. The method of claim 9, further comprising sorting the non-motile sperm cells into a plurality of aliquots. 11. The method of claim 10, wherein each of the plurality of aliquots comprises from about 1 to about 20 non-motile sperm cells. 12. The method of claim 11, wherein each of the plurality of aliquots comprises from about 1 to about 10 non-motile sperm cells. 13. The method of claim 12, wherein each of the plurality of aliquots comprises a single non-motile sperm cell. 14. A tissue sample processing system, comprising:
a microfluidic separating system having
a fluid channel to receive a sample fluid including a carrier fluid and a tissue sample, and
a plurality of outlets, wherein flow of the carrier fluid and the tissue sample in the fluid channel facilitates segregation of materials in the tissue sample based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the tissue sample; and
a cryopreservation system associated with at least one of the plurality of outlets to freeze the material in the tissue sample associated with the at least one of the plurality of outlets. 15. The system of claim 14, wherein the fluid channel comprises a spiral configuration. 16. The system of claim 14, wherein the microfluidic separating system comprises:
an inlet zone having a carrier fluid inlet to receive the carrier fluid and a tissue sample inlet to receive the tissue sample; an outlet zone having the plurality of outlets; and a transport region between the inlet zone and the outlet zone, the transport region being open to the carrier fluid and the tissue sample, wherein a cross-flow in the transport region facilitates segregation of the materials in the tissue sample based on size. 17. The system of claim 14, wherein the tissue sample comprises a sperm sample. 18. The system of claim 14, wherein the material in the tissue sample associated with the at least one of the plurality of outlets comprises non-motile sperm cells. 19. A tissue sample processing system, comprising:
a microfluidic separating system having
a fluid channel to receive a carrier fluid and a tissue sample, and
a plurality of outlets, wherein flow of the carrier fluid and the tissue sample in the fluid channel facilitates segregation of materials in the tissue sample based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the tissue sample; and
a sorting system associated with at least one of the plurality of outlets to sort a plurality of aliquots of the material in the tissue sample associated with the at least one of the plurality of outlets. 20. The system of claim 19, wherein the sorting system comprises:
an inlet to receive the material in the tissue sample associated with the at least one of the plurality of outlets; a plurality of aliquot storage channels; and a plurality of valves associated with the plurality of aliquot storage channels to facilitate separation of the plurality of aliquots into the aliquot storage channels. 21. The system of claim 19, wherein the fluid channel comprises a spiral configuration. 22. The system of claim 19, wherein the microfluidic separating system comprises:
an inlet zone having a carrier fluid inlet to receive the carrier fluid and a tissue sample inlet to receive the tissue sample; an outlet zone having the plurality of outlets; and a transport region between the inlet zone and the outlet zone, the transport region being open to the carrier fluid and the tissue sample, wherein a cross-flow in the transport region facilitates segregation of the materials in the tissue sample based on size. 23. The system of claim 19, further comprising a cryopreservation system associated with the sorting system to freeze at least one of the plurality of aliquots of the material in the tissue sample. 24. The system of claim 19, wherein each of the plurality of aliquots comprises from about 1 to about 20 non-motile sperm cells. 25. The system of claim 24, wherein each of the plurality of aliquots comprises from about 1 to about 10 non-motile sperm cells. 26. The system of claim 25, wherein each of the plurality of aliquots comprises a single non-motile sperm cell. 27. A tissue sample processing system, comprising:
a microfluidic separating system having
a fluid channel to receive a carrier fluid and a tissue sample, and
a plurality of outlets, wherein flow of the carrier fluid and the tissue sample in the fluid channel facilitates segregation of materials in the tissue sample based on size into a plurality of size fractions, such that each one of the plurality of outlets receives a different size fraction of the materials in the tissue sample; and
a concentrating system associated with at least one of the plurality of outlets to concentrate a size fraction of the material in the tissue sample associated with the at least one of the plurality of outlets. 28. The system of claim 27, wherein the concentrating system comprises a first fluid conduit and a second fluid conduit, wherein the first fluid conduit is sized to prevent passage of the size fraction of the material therethrough, and wherein a fluid resistance of the first fluid conduit is greater than a fluid resistance of the second fluid conduit such that the size fraction of material flows towards the first conduit and is trapped at the first conduit. 29. The system of claim 27, wherein the fluid channel comprises a spiral configuration. 30. The system of claim 27, wherein the microfluidic separating system comprises:
an inlet zone having a carrier fluid inlet to receive the carrier fluid and a tissue sample inlet to receive the tissue sample; an outlet zone having the plurality of outlets; and a transport region between the inlet zone and the outlet zone, the transport region being open to the carrier fluid and the tissue sample, wherein a cross-flow in the transport region facilitates segregation of the materials in the tissue sample based on size. 31. The system of claim 27, further comprising a cryopreservation system associated with the concentrating system to freeze the concentrated size fraction of the material in the tissue sample. 32. The system of claim 27, wherein the tissue sample comprises a sperm sample. | 1,600 |
1,076 | 14,639,162 | 1,633 | A therapeutic patch includes at least one layer of a nanofiber fabric and at least one of a plurality of stem cells or a plurality of stem cell-derived paracrine factors embedded in the nanofiber fabric. The therapeutic patch is produced such that the nanofiber fabric is formed of a nanofiber web. | 1. A therapeutic patch, comprising:
at least one layer of a nanofiber fabric; and at least one of a plurality of stem cells and a plurality of stem cell-derived paracrine factors embedded in the nanofiber fabric,
wherein the nanofiber fabric comprises a nanofiber web. 2. The therapeutic patch according to claim 1, wherein the nanofiber fabric comprises a nonwoven nanofiber fabric having a fiber diameter from about 80 nm to about 2 μm. 3. The therapeutic patch according to claim 1, wherein the nanofiber fabric comprises a plurality of pores, said plurality of pores comprising a maximal pore size. 4. The therapeutic patch according to claim 3, wherein the embedded stem cells comprise an embedded stem cell diameter, and
wherein the maximal pore size is less than the embedded stem cell diameter, the maximal pore size is less than other cell diameters, and the maximal pore size is greater than embedded paracrine factor, cell product, nutrient, or waste diameters. 5. The therapeutic patch according to claim 1, wherein the nanofiber fabric comprises a biodegradable polymer. 6. The therapeutic patch according to claim 4, wherein the nanofiber fabric comprises at least one of a synthetic biocompatible material, a biological material, and any combination thereof. 7. The therapeutic patch according to claim 5, wherein the nanofiber fabric comprises at least one of poly(L-lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, polyethylene oxide, poly(ethylene terephthalate), poly(vinyl alcohol), collagen, and any combination thereof. 8. A method of treatment, comprising:
preparing a therapeutic patch comprising:
at least one layer of a nanofiber fabric; and
at least one of a plurality of stem cells or a plurality of stem cell-derived paracrine factors embedded in the nanofiber fabric; and
applying the therapeutic patch to an object,
wherein the nanofiber fabric comprises a nanofiber web. 9. The method of treatment according to claim 8, wherein the nanofiber fabric comprises a nonwoven nanofiber fabric having a fiber diameter from about 80 nm to about 2 μm. 10. The method of treatment according to claim 8, wherein the nanofiber fabric comprises a plurality of pores, said plurality of pores comprising a maximal pore size. 11. The method of treatment according to claim 10, wherein the embedded stem cells comprise an embedded stem cell diameter, and
wherein the maximal pore size is less than the embedded stem cell diameter, the maximal pore size is less than other cell diameters, and the maximal pore size is greater than embedded paracrine factor, cell product, nutrient, or waste diameters. 12. The method of treatment according to claim 8, wherein the nanofiber fabric comprises a biodegradable polymer. 13. The method of treatment according to claim 11, wherein the nanofiber fabric comprises at least one synthetic biocompatible material, a biological material, and any combination thereof. 14. The method of treatment according to claim 12, wherein the nanofiber fabric comprises at least one of poly(L-lactic acid), poly(lactic-co-glycolic acid) copolymer, polycaprolactone, poly(ethylene oxide), poly(ethylene terephthalate), poly(vinyl alcohol), collagen, and any combination thereof. 15. The method of treatment according to claim 8, wherein applying the therapeutic patch to an object comprises applying the therapeutic patch to at least one of a tissue, an organ or portion of an organ, a blood vessel, a muscle, a bone, a joint, a bypass graft, a medical device, and any combination thereof. 16. The method of treatment according to claim 15, wherein applying the therapeutic patch to an object comprises applying the therapeutic patch to a stent, a pacemaker, an implantable cardioverter-defibrillator, a pacemaker lead, an implantable cardioverter-defibrillator lead, a biventricular implantable cardioverter-defibrillator lead, an artificial heart, an artificial valve, a ventricular assist device, a balloon pump, a catheter, a central venous line, an orthopedic implant, wound healing packing, and wound healing gauze. 17. The method of treatment according to claim 8, wherein preparing a therapeutic patch comprises preparing a single-use therapeutic patch including stem cell-derived paracrine factors. 18. The method of treatment according to claim 17, wherein preparing the single-use therapeutic patch comprises at least one of directly absorbing conditioned media into the therapeutic patch, immersing the therapeutic patch in a homogenized cell solution, and lysing stem cells in the therapeutic patch. 19. The method of treatment according to claim 8, wherein preparing a therapeutic patch comprises preparing a reusable therapeutic patch including stem cells. 20. The method of treatment according to claim 19, wherein preparing the reusable therapeutic patch comprises:
seeding stem cells into the therapeutic patch; and incubating the stem cells in the therapeutic patch. | A therapeutic patch includes at least one layer of a nanofiber fabric and at least one of a plurality of stem cells or a plurality of stem cell-derived paracrine factors embedded in the nanofiber fabric. The therapeutic patch is produced such that the nanofiber fabric is formed of a nanofiber web.1. A therapeutic patch, comprising:
at least one layer of a nanofiber fabric; and at least one of a plurality of stem cells and a plurality of stem cell-derived paracrine factors embedded in the nanofiber fabric,
wherein the nanofiber fabric comprises a nanofiber web. 2. The therapeutic patch according to claim 1, wherein the nanofiber fabric comprises a nonwoven nanofiber fabric having a fiber diameter from about 80 nm to about 2 μm. 3. The therapeutic patch according to claim 1, wherein the nanofiber fabric comprises a plurality of pores, said plurality of pores comprising a maximal pore size. 4. The therapeutic patch according to claim 3, wherein the embedded stem cells comprise an embedded stem cell diameter, and
wherein the maximal pore size is less than the embedded stem cell diameter, the maximal pore size is less than other cell diameters, and the maximal pore size is greater than embedded paracrine factor, cell product, nutrient, or waste diameters. 5. The therapeutic patch according to claim 1, wherein the nanofiber fabric comprises a biodegradable polymer. 6. The therapeutic patch according to claim 4, wherein the nanofiber fabric comprises at least one of a synthetic biocompatible material, a biological material, and any combination thereof. 7. The therapeutic patch according to claim 5, wherein the nanofiber fabric comprises at least one of poly(L-lactic acid), poly(lactic-co-glycolic acid), polycaprolactone, polyethylene oxide, poly(ethylene terephthalate), poly(vinyl alcohol), collagen, and any combination thereof. 8. A method of treatment, comprising:
preparing a therapeutic patch comprising:
at least one layer of a nanofiber fabric; and
at least one of a plurality of stem cells or a plurality of stem cell-derived paracrine factors embedded in the nanofiber fabric; and
applying the therapeutic patch to an object,
wherein the nanofiber fabric comprises a nanofiber web. 9. The method of treatment according to claim 8, wherein the nanofiber fabric comprises a nonwoven nanofiber fabric having a fiber diameter from about 80 nm to about 2 μm. 10. The method of treatment according to claim 8, wherein the nanofiber fabric comprises a plurality of pores, said plurality of pores comprising a maximal pore size. 11. The method of treatment according to claim 10, wherein the embedded stem cells comprise an embedded stem cell diameter, and
wherein the maximal pore size is less than the embedded stem cell diameter, the maximal pore size is less than other cell diameters, and the maximal pore size is greater than embedded paracrine factor, cell product, nutrient, or waste diameters. 12. The method of treatment according to claim 8, wherein the nanofiber fabric comprises a biodegradable polymer. 13. The method of treatment according to claim 11, wherein the nanofiber fabric comprises at least one synthetic biocompatible material, a biological material, and any combination thereof. 14. The method of treatment according to claim 12, wherein the nanofiber fabric comprises at least one of poly(L-lactic acid), poly(lactic-co-glycolic acid) copolymer, polycaprolactone, poly(ethylene oxide), poly(ethylene terephthalate), poly(vinyl alcohol), collagen, and any combination thereof. 15. The method of treatment according to claim 8, wherein applying the therapeutic patch to an object comprises applying the therapeutic patch to at least one of a tissue, an organ or portion of an organ, a blood vessel, a muscle, a bone, a joint, a bypass graft, a medical device, and any combination thereof. 16. The method of treatment according to claim 15, wherein applying the therapeutic patch to an object comprises applying the therapeutic patch to a stent, a pacemaker, an implantable cardioverter-defibrillator, a pacemaker lead, an implantable cardioverter-defibrillator lead, a biventricular implantable cardioverter-defibrillator lead, an artificial heart, an artificial valve, a ventricular assist device, a balloon pump, a catheter, a central venous line, an orthopedic implant, wound healing packing, and wound healing gauze. 17. The method of treatment according to claim 8, wherein preparing a therapeutic patch comprises preparing a single-use therapeutic patch including stem cell-derived paracrine factors. 18. The method of treatment according to claim 17, wherein preparing the single-use therapeutic patch comprises at least one of directly absorbing conditioned media into the therapeutic patch, immersing the therapeutic patch in a homogenized cell solution, and lysing stem cells in the therapeutic patch. 19. The method of treatment according to claim 8, wherein preparing a therapeutic patch comprises preparing a reusable therapeutic patch including stem cells. 20. The method of treatment according to claim 19, wherein preparing the reusable therapeutic patch comprises:
seeding stem cells into the therapeutic patch; and incubating the stem cells in the therapeutic patch. | 1,600 |
1,077 | 15,189,839 | 1,656 | The present invention provides a method of modulating various aspects of the immune system. In particular, the present invention teaches the use of diketopiperazines (DKPs) to modulate various aspects of the immune system such as, for example, inflammation, T-cells and various cytokines. | 1. A method of inhibiting inflammation, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising a low molecular weight fraction (LMWF) of human serum albumin (HSA), wherein the individual is not administered a cyclooxygenase-2 (COX-2) antagonist within the period of time ranging from about one hour before to about 6 hours after administration of the pharmaceutical composition. 2. The method of claim 1, wherein the LMWF of HSA contains components having a molecular weight of less than 5000. 3. The method of claim 1, wherein the LMWF of HSA contains components having a molecular weight less than 3000. 4. The method of claim 1, wherein at least one of the components in the LMWF of HSA comprises DA-DKP. 5. The method of claim 1, wherein the LMWF of HSA comprises one or more compounds selected from the group consisting of N-acetyl tryptophan and caprylic acid. 6. The method of claim 1, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least about one hour, at least about 12 hours, at least about one day, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at least about seven days, at least about eight days or at least about nine days and 10 days, prior to administration of the pharmaceutical composition. 7. The method of claim 1, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist during the period of time in which at least one active ingredient in the pharmaceutical composition exerts its effect. 8. The method of claim 1, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least 6 hours, at least 12 hours, at least about 24 hours, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at last about one week, at least about two weeks, at least about three weeks, at least about one month, at least about two months, at least about three months, at least about four months at least about five months or at least about six months after administration of a pharmaceutical composition of the invention. 9. The method of claim 7, wherein the composition that reduces COX-2 activity comprises a chemical selected from the group consisting of acetylsalicylic acid (aspirin), 2-(4-isobutylphenyl)propanoic acid (ibuprofen), N-(4-hydroxyphenyl)ethanamide (paracetamol), (S)-6-methoxy-α-methyl-2-naphthaleneacetic acid (naproxen), 2-[(2,6-dichlorophenyl)amino] benzeneacetic acid (diclofenac), 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (celecoxib), 4-[4-(methylsulfonyl)phenyl]-3-phenyl-2(5H)-furanone (rofecoxib), and 4-(5-Methyl-3-phenylisoxazol-4-yl)benzolsulfonamid (valdecoxib). 10. A method of treating a T-cell mediated disease, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising a low molecular weight fraction (LMWF) of human serum albumin (HSA), wherein the individual is not administered a cyclooxygenase-2 (COX-2) antagonist within the period of time ranging from about one hour before to about six hours after administration of the pharmaceutical composition. 11. The method of claim 10, wherein the T-cell mediated disease is graft rejection, graft versus host disease, an unwanted delayed-type hypersensitivity reaction, a T-cell mediated pulmonary disease, an autoimmune disease or an inflammatory disease. 12. The method of claim 10, wherein the T-cell-mediated disease is selected form the group consisting of multiple sclerosis, neuritis, polymyositis, psoriasis, vitiligo, Sjogren's syndrome, rheumatoid arthritis, Type 1 diabetes, autoimmune pancreatitis, inflammatory bowel diseases, Crohn's disease, ulcerative colitis, celiac disease, glomerulonephritis, scleroderma, sarcoidosis, autoimmune thyroid diseases, Hashimoto's thyroiditis, Graves disease, myasthenia gravis, Addison's disease, autoimmune uveoretinitis, pemphigus vulgaris, primary biliary cirrhosis, pernicious anemia and systemic lupus erythematosis. 13. The method of claim 10, wherein the T-cell-mediated disease is pulmonary fibrosis or idiopathic pulmonary fibrosis. 14. The method of claim 10, wherein the T-cell-mediated disease is an inflammatory disease. 15. The method of claim 10, wherein the LMWF of HSA contains components having a molecular weight of less than 5000. 16. The method of claim 10, wherein the LMWF of HSA contains components having a molecular weight less than 3000. 17. The method of claim 10, wherein at least one of the components in the LMWF of HSA comprises DA-DKP. 18. The method of claim 10, wherein the LMWF of HSA comprises one or more compounds selected from the group consisting of N-acetyl tryptophan and caprylic acid. 19. The method of claim 10, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least about one hour, at least about 12 hours, at least about one day, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at least about seven days, at least about eight days or at least about nine days and 10 days, prior to administration of the pharmaceutical composition. 20. The method of claim 10, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist during the period of time in which at least one active ingredient in the pharmaceutical composition exerts its effect. 21. The method of claim 10, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least 6 hours, at least 12 hours, at least about 24 hours, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at last about one week, at least about two weeks, at least about three weeks, at least about one month, at least about two months, at least about three months, at least about four months at least about five months or at least about six months after administration of a pharmaceutical composition of the invention. 22. The method of claim 20, wherein the composition that reduces COX-2 activity comprises a chemical selected from the group consisting of acetylsalicylic acid (aspirin), 2-(4-isobutylphenyl)propanoic acid (ibuprofen), N-(4-hydroxyphenyl)ethanamide (paracetamol), (S)-6-methoxy-α-methyl-2-naphthaleneacetic acid (naproxen), 2-[(2,6-dichlorophenyl)amino] benzeneacetic acid (diclofenac), 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (celecoxib), 4-[4-(methylsulfonyl)phenyl]-3-phenyl-2(5H)-furanone (rofecoxib), and 4-(5-Methyl-3-phenylisoxazol-4-yl)benzolsulfonamid (valdecoxib). 23. A method of treating an individual for a joint disease or condition, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising a LMWF of HSA, wherein the individual is not administered a COX-2 antagonist within the period of time ranging from about one hour before to about six hours after administration of the pharmaceutical composition. 24. The method of claim 23, wherein the joint disease or condition is a degenerative joint disease. 25. The method of claim 23, wherein the LMWF of HSA contains components having a molecular weight of less than 5000. 26. The method of claim 23, wherein the LMWF of HSA contains components having a molecular weight less than 3000. 27. The method of claim 23, wherein at least one of the components in the LMWF of HSA comprises DA-DKP. 28. The method of claim 23, wherein the LMWF of HSA comprises one or more compounds selected from the group consisting of N-acetyl tryptophan and caprylic acid. 29. The method of claim 23, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least about one hour, at least about 12 hours, at least about one day, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at least about seven days, at least about eight days or at least about nine days and 10 days, prior to administration of the pharmaceutical composition. 30. The method of claim 23, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist during the period of time in which at least one active ingredient in the pharmaceutical composition exerts its effect. 31. The method of claim 23, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least 6 hours, at least 12 hours, at least about 24 hours, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at last about one week, at least about two weeks, at least about three weeks, at least about one month, at least about two months, at least about three months, at least about four months at least about five months or at least about six months after administration of a pharmaceutical composition of the invention. 32. The method of claim 30, wherein the composition that reduces COX-2 activity comprises a chemical selected from the group consisting of acetylsalicylic acid (aspirin), 2-(4-isobutylphenyl)propanoic acid (ibuprofen), N-(4-hydroxyphenyl)ethanamide (paracetamol), (S)-6-methoxy-α-methyl-2-naphthaleneacetic acid (naproxen), 2-[(2,6-dichlorophenyl)amino] benzeneacetic acid (diclofenac), 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (celecoxib), 4-[4-(methylsulfonyl)phenyl]-3-phenyl-2(5H)-furanone (rofecoxib), and 4-(5-Methyl-3-phenylisoxazol-4-yl)benzolsulfonamid (valdecoxib). 33. The method of reducing the level of IL-8 in an individual, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising a LMWF of HSA, wherein the individual is not administered a COX-2 antagonist within the period of time ranging from about one hour before to about six hours after administration of the pharmaceutical composition. | The present invention provides a method of modulating various aspects of the immune system. In particular, the present invention teaches the use of diketopiperazines (DKPs) to modulate various aspects of the immune system such as, for example, inflammation, T-cells and various cytokines.1. A method of inhibiting inflammation, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising a low molecular weight fraction (LMWF) of human serum albumin (HSA), wherein the individual is not administered a cyclooxygenase-2 (COX-2) antagonist within the period of time ranging from about one hour before to about 6 hours after administration of the pharmaceutical composition. 2. The method of claim 1, wherein the LMWF of HSA contains components having a molecular weight of less than 5000. 3. The method of claim 1, wherein the LMWF of HSA contains components having a molecular weight less than 3000. 4. The method of claim 1, wherein at least one of the components in the LMWF of HSA comprises DA-DKP. 5. The method of claim 1, wherein the LMWF of HSA comprises one or more compounds selected from the group consisting of N-acetyl tryptophan and caprylic acid. 6. The method of claim 1, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least about one hour, at least about 12 hours, at least about one day, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at least about seven days, at least about eight days or at least about nine days and 10 days, prior to administration of the pharmaceutical composition. 7. The method of claim 1, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist during the period of time in which at least one active ingredient in the pharmaceutical composition exerts its effect. 8. The method of claim 1, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least 6 hours, at least 12 hours, at least about 24 hours, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at last about one week, at least about two weeks, at least about three weeks, at least about one month, at least about two months, at least about three months, at least about four months at least about five months or at least about six months after administration of a pharmaceutical composition of the invention. 9. The method of claim 7, wherein the composition that reduces COX-2 activity comprises a chemical selected from the group consisting of acetylsalicylic acid (aspirin), 2-(4-isobutylphenyl)propanoic acid (ibuprofen), N-(4-hydroxyphenyl)ethanamide (paracetamol), (S)-6-methoxy-α-methyl-2-naphthaleneacetic acid (naproxen), 2-[(2,6-dichlorophenyl)amino] benzeneacetic acid (diclofenac), 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (celecoxib), 4-[4-(methylsulfonyl)phenyl]-3-phenyl-2(5H)-furanone (rofecoxib), and 4-(5-Methyl-3-phenylisoxazol-4-yl)benzolsulfonamid (valdecoxib). 10. A method of treating a T-cell mediated disease, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising a low molecular weight fraction (LMWF) of human serum albumin (HSA), wherein the individual is not administered a cyclooxygenase-2 (COX-2) antagonist within the period of time ranging from about one hour before to about six hours after administration of the pharmaceutical composition. 11. The method of claim 10, wherein the T-cell mediated disease is graft rejection, graft versus host disease, an unwanted delayed-type hypersensitivity reaction, a T-cell mediated pulmonary disease, an autoimmune disease or an inflammatory disease. 12. The method of claim 10, wherein the T-cell-mediated disease is selected form the group consisting of multiple sclerosis, neuritis, polymyositis, psoriasis, vitiligo, Sjogren's syndrome, rheumatoid arthritis, Type 1 diabetes, autoimmune pancreatitis, inflammatory bowel diseases, Crohn's disease, ulcerative colitis, celiac disease, glomerulonephritis, scleroderma, sarcoidosis, autoimmune thyroid diseases, Hashimoto's thyroiditis, Graves disease, myasthenia gravis, Addison's disease, autoimmune uveoretinitis, pemphigus vulgaris, primary biliary cirrhosis, pernicious anemia and systemic lupus erythematosis. 13. The method of claim 10, wherein the T-cell-mediated disease is pulmonary fibrosis or idiopathic pulmonary fibrosis. 14. The method of claim 10, wherein the T-cell-mediated disease is an inflammatory disease. 15. The method of claim 10, wherein the LMWF of HSA contains components having a molecular weight of less than 5000. 16. The method of claim 10, wherein the LMWF of HSA contains components having a molecular weight less than 3000. 17. The method of claim 10, wherein at least one of the components in the LMWF of HSA comprises DA-DKP. 18. The method of claim 10, wherein the LMWF of HSA comprises one or more compounds selected from the group consisting of N-acetyl tryptophan and caprylic acid. 19. The method of claim 10, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least about one hour, at least about 12 hours, at least about one day, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at least about seven days, at least about eight days or at least about nine days and 10 days, prior to administration of the pharmaceutical composition. 20. The method of claim 10, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist during the period of time in which at least one active ingredient in the pharmaceutical composition exerts its effect. 21. The method of claim 10, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least 6 hours, at least 12 hours, at least about 24 hours, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at last about one week, at least about two weeks, at least about three weeks, at least about one month, at least about two months, at least about three months, at least about four months at least about five months or at least about six months after administration of a pharmaceutical composition of the invention. 22. The method of claim 20, wherein the composition that reduces COX-2 activity comprises a chemical selected from the group consisting of acetylsalicylic acid (aspirin), 2-(4-isobutylphenyl)propanoic acid (ibuprofen), N-(4-hydroxyphenyl)ethanamide (paracetamol), (S)-6-methoxy-α-methyl-2-naphthaleneacetic acid (naproxen), 2-[(2,6-dichlorophenyl)amino] benzeneacetic acid (diclofenac), 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (celecoxib), 4-[4-(methylsulfonyl)phenyl]-3-phenyl-2(5H)-furanone (rofecoxib), and 4-(5-Methyl-3-phenylisoxazol-4-yl)benzolsulfonamid (valdecoxib). 23. A method of treating an individual for a joint disease or condition, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising a LMWF of HSA, wherein the individual is not administered a COX-2 antagonist within the period of time ranging from about one hour before to about six hours after administration of the pharmaceutical composition. 24. The method of claim 23, wherein the joint disease or condition is a degenerative joint disease. 25. The method of claim 23, wherein the LMWF of HSA contains components having a molecular weight of less than 5000. 26. The method of claim 23, wherein the LMWF of HSA contains components having a molecular weight less than 3000. 27. The method of claim 23, wherein at least one of the components in the LMWF of HSA comprises DA-DKP. 28. The method of claim 23, wherein the LMWF of HSA comprises one or more compounds selected from the group consisting of N-acetyl tryptophan and caprylic acid. 29. The method of claim 23, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least about one hour, at least about 12 hours, at least about one day, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at least about seven days, at least about eight days or at least about nine days and 10 days, prior to administration of the pharmaceutical composition. 30. The method of claim 23, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist during the period of time in which at least one active ingredient in the pharmaceutical composition exerts its effect. 31. The method of claim 23, wherein the individual receiving the pharmaceutical composition is not administered a COX-2 antagonist within a time period selected from the group consisting of at least 6 hours, at least 12 hours, at least about 24 hours, at least about two days, at least about three days, at least about four days, at least about five days, at least about six days, at last about one week, at least about two weeks, at least about three weeks, at least about one month, at least about two months, at least about three months, at least about four months at least about five months or at least about six months after administration of a pharmaceutical composition of the invention. 32. The method of claim 30, wherein the composition that reduces COX-2 activity comprises a chemical selected from the group consisting of acetylsalicylic acid (aspirin), 2-(4-isobutylphenyl)propanoic acid (ibuprofen), N-(4-hydroxyphenyl)ethanamide (paracetamol), (S)-6-methoxy-α-methyl-2-naphthaleneacetic acid (naproxen), 2-[(2,6-dichlorophenyl)amino] benzeneacetic acid (diclofenac), 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (celecoxib), 4-[4-(methylsulfonyl)phenyl]-3-phenyl-2(5H)-furanone (rofecoxib), and 4-(5-Methyl-3-phenylisoxazol-4-yl)benzolsulfonamid (valdecoxib). 33. The method of reducing the level of IL-8 in an individual, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising a LMWF of HSA, wherein the individual is not administered a COX-2 antagonist within the period of time ranging from about one hour before to about six hours after administration of the pharmaceutical composition. | 1,600 |
1,078 | 15,322,003 | 1,649 | The present invention provides a composition for treating ischemic diseases or neuroinflammatory diseases. PSA-NCAM-positive neural progenitor cells used in the present invention promote angiogenesis in injected tissue and inhibit an inflammatory response. The PSA-NCAM-positive neural progenitor cells can be simply isolated by using an anti-PSA-NCAM-antibody, and exhibit excellent angiogenic and anti-inflammatory activities compared with mesenchymal stem cells, and thus can be useful as a composition for effectively treating ischemic diseases caused by a vascular injury and nerve damage diseases caused by inflammation. In addition, a secretome of the neural progenitor cells of the present invention reduces the ischemic injury site and allows a neurological function to recover, and thus can be used as an agent for treating ischemic diseases and degenerative nervous system disorders such as nerve damage diseases caused by inflammation. | 1-18. (canceled) 19. A method for treating ischemic disease or neuroinflammatory disease, the method comprising administering a composition containing poly-sialylated neural cell adhesion molecule (PSA-NCAM)-positive neural precursor cells as an active ingredient to a subject in need thereof. 20. A method for treating ischemic disease or neuroinflammatory disease, the method comprising administering a composition containing, as an active ingredient, a secretome of neural precursor cells to a subject in need thereof. 21. The method of claim 19, wherein the composition increases the expression of angiopoietin-1. 22. The method of claim 19, wherein the composition inhibits the activation of glial cells or astrocytes. 23. The method of claim 22, wherein the composition reduces the expression of CD68 or GFAP. 24. The method of claim 19, wherein the PSA-NCAM-positive neural precursor cells are separated from neural rosettes differentiated from pluripotent stem cells. 25. The method of claim 20, wherein the neural precursor cells are differentiated from pluripotent stem cells. 26. The method of claim 20, wherein the neural precursor cells are neural precursor cells at the stage of neural rosettes differentiated from pluripotent stem cells. 27. The method of claim 20, wherein the neural precursor cells are poly-sialylated neural cell adhesion molecule (PSA-NCAM)-positive neural precursor cells. 28. The method of claim 20, wherein the neural precursor cells are poly-sialylated neural cell adhesion molecule (PSA-NCAM)-negative neural precursor cells. 29. The method of claim 20, wherein the secretome is in a form of being contained in a cell culture liquid obtained by culturing neural precursor cells in an animal cell culture medium. 30. The method of claim 29, wherein the cell culture liquid is obtained by culturing the neural precursor cells in a serum-free animal cell culture medium containing insulin/transferrin/selenium (ITS) and basic fibroblast growth factor (bFGT) and then removing the cells. 31. The method of claim 20, wherein the secretome comprises the following proteins:
Agrin, annexin A5, BSG (Basigin), biglycan, calponin-3, coactosin-like protein, cofilin-1, collagen alpha-2, cullin-3, destrin, dystroglycan, ephrin-B2, exportin-2, ezrin, fibronectin, fibulin-1, frizzled-related protein, gelatin-3 binding protein, granulins, growth/differentiation factor 11, haptoglobin, hemopexin, high mobility group protein B2, hornerin, importin-9, insulin-like growth factor-binding protein 2, Lupus La protein, macrophage migration inhibitory factor, midkine, moesin, neuropilin 2, pleiotrophin, profilin-1, protein DJ-1, radixin, secreted frizzled-related protein-2, septin-11, talin-1, testican, thymopoietin, transgelin-3 and vimentin. 32. The method of claim 20, wherein the secretome comprises the following proteins:
Agrin, annexin A2, attractin, biglycan, ceruloplasmin, cofilin-1, collagen alpha-1, coronin-1X, dermicidin, DERP12, eprin-B3, exostosin-2, ezrin, gelatin-3 binding protein, granulins, growth/differentiation factor 11, haptoglobin, hemopexin, high mobility group protein B2, hornerin, insulin-like growth factor-binding protein 2, Lupus La protein, midkine, moesin, multiple epidermal growth factor-like domains protein 8, nidogen-1, parathymosin, profilin-2, protein DJ-1, secreted frizzled-related protein-2, secretogranin, talin-1, thymosin beta-4, TGFBI (Transforming growth factor-beta-induced protein ig-h3), transgelin and vimentin. 33. The method of claim 19 or 20, wherein the ischemic disease is selected from the group consisting of ischemic cerebrovascular disease, ischemic heart disease, myocardial infarction, angina pectoris, lower limb artery ischemic disease, and distal limb ischemic disease. 34. The method of claim 33, wherein the ischemic cerebrovascular disease is ischemic stroke. 35. The method of claim 19 or 20, wherein the neuroinflammatory disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, Huntington's disease, Lou Gehrig's disease, Creutzfeldt Jakob disease, multiple sclerosis, amyotrophic lateral sclerosis, diffuse Lewy body disease, leukencephalitis, temporal lobe epilepsy, and inflammatory spinal cord injury. 36. The method of claim 24, 25, or 26, wherein the pluripotent stem cells are embryonic stem cells, induced pluripotent stem cells (iPSCs), embryonic germ cells, or embryonic carcinoma cells. | The present invention provides a composition for treating ischemic diseases or neuroinflammatory diseases. PSA-NCAM-positive neural progenitor cells used in the present invention promote angiogenesis in injected tissue and inhibit an inflammatory response. The PSA-NCAM-positive neural progenitor cells can be simply isolated by using an anti-PSA-NCAM-antibody, and exhibit excellent angiogenic and anti-inflammatory activities compared with mesenchymal stem cells, and thus can be useful as a composition for effectively treating ischemic diseases caused by a vascular injury and nerve damage diseases caused by inflammation. In addition, a secretome of the neural progenitor cells of the present invention reduces the ischemic injury site and allows a neurological function to recover, and thus can be used as an agent for treating ischemic diseases and degenerative nervous system disorders such as nerve damage diseases caused by inflammation.1-18. (canceled) 19. A method for treating ischemic disease or neuroinflammatory disease, the method comprising administering a composition containing poly-sialylated neural cell adhesion molecule (PSA-NCAM)-positive neural precursor cells as an active ingredient to a subject in need thereof. 20. A method for treating ischemic disease or neuroinflammatory disease, the method comprising administering a composition containing, as an active ingredient, a secretome of neural precursor cells to a subject in need thereof. 21. The method of claim 19, wherein the composition increases the expression of angiopoietin-1. 22. The method of claim 19, wherein the composition inhibits the activation of glial cells or astrocytes. 23. The method of claim 22, wherein the composition reduces the expression of CD68 or GFAP. 24. The method of claim 19, wherein the PSA-NCAM-positive neural precursor cells are separated from neural rosettes differentiated from pluripotent stem cells. 25. The method of claim 20, wherein the neural precursor cells are differentiated from pluripotent stem cells. 26. The method of claim 20, wherein the neural precursor cells are neural precursor cells at the stage of neural rosettes differentiated from pluripotent stem cells. 27. The method of claim 20, wherein the neural precursor cells are poly-sialylated neural cell adhesion molecule (PSA-NCAM)-positive neural precursor cells. 28. The method of claim 20, wherein the neural precursor cells are poly-sialylated neural cell adhesion molecule (PSA-NCAM)-negative neural precursor cells. 29. The method of claim 20, wherein the secretome is in a form of being contained in a cell culture liquid obtained by culturing neural precursor cells in an animal cell culture medium. 30. The method of claim 29, wherein the cell culture liquid is obtained by culturing the neural precursor cells in a serum-free animal cell culture medium containing insulin/transferrin/selenium (ITS) and basic fibroblast growth factor (bFGT) and then removing the cells. 31. The method of claim 20, wherein the secretome comprises the following proteins:
Agrin, annexin A5, BSG (Basigin), biglycan, calponin-3, coactosin-like protein, cofilin-1, collagen alpha-2, cullin-3, destrin, dystroglycan, ephrin-B2, exportin-2, ezrin, fibronectin, fibulin-1, frizzled-related protein, gelatin-3 binding protein, granulins, growth/differentiation factor 11, haptoglobin, hemopexin, high mobility group protein B2, hornerin, importin-9, insulin-like growth factor-binding protein 2, Lupus La protein, macrophage migration inhibitory factor, midkine, moesin, neuropilin 2, pleiotrophin, profilin-1, protein DJ-1, radixin, secreted frizzled-related protein-2, septin-11, talin-1, testican, thymopoietin, transgelin-3 and vimentin. 32. The method of claim 20, wherein the secretome comprises the following proteins:
Agrin, annexin A2, attractin, biglycan, ceruloplasmin, cofilin-1, collagen alpha-1, coronin-1X, dermicidin, DERP12, eprin-B3, exostosin-2, ezrin, gelatin-3 binding protein, granulins, growth/differentiation factor 11, haptoglobin, hemopexin, high mobility group protein B2, hornerin, insulin-like growth factor-binding protein 2, Lupus La protein, midkine, moesin, multiple epidermal growth factor-like domains protein 8, nidogen-1, parathymosin, profilin-2, protein DJ-1, secreted frizzled-related protein-2, secretogranin, talin-1, thymosin beta-4, TGFBI (Transforming growth factor-beta-induced protein ig-h3), transgelin and vimentin. 33. The method of claim 19 or 20, wherein the ischemic disease is selected from the group consisting of ischemic cerebrovascular disease, ischemic heart disease, myocardial infarction, angina pectoris, lower limb artery ischemic disease, and distal limb ischemic disease. 34. The method of claim 33, wherein the ischemic cerebrovascular disease is ischemic stroke. 35. The method of claim 19 or 20, wherein the neuroinflammatory disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, Huntington's disease, Lou Gehrig's disease, Creutzfeldt Jakob disease, multiple sclerosis, amyotrophic lateral sclerosis, diffuse Lewy body disease, leukencephalitis, temporal lobe epilepsy, and inflammatory spinal cord injury. 36. The method of claim 24, 25, or 26, wherein the pluripotent stem cells are embryonic stem cells, induced pluripotent stem cells (iPSCs), embryonic germ cells, or embryonic carcinoma cells. | 1,600 |
1,079 | 15,840,046 | 1,612 | A two-phase oral care whitening composition including a hydrophobic phase and a hydrophilic phase, and methods for oral care whitening teeth with the same are provided. The hydrophobic phase may include a source of hydrogen peroxide and an acyl donor. The hydrophilic phase may include an enzyme that catalyzes the generation of peracetic acid between the source of hydrogen peroxide and the acyl donor. At least one of the hydrophobic phase and the hydrophilic phase may further include at least one surfactant. | 1. A two-phase oral care whitening composition, comprising:
a hydrophobic phase comprising a source of hydrogen peroxide and an acyl donor; and a hydrophilic phase comprising an enzyme that catalyzes the generation of peracetic acid between the source of hydrogen peroxide and the acyl donor, wherein at least one of the hydrophobic phase and the hydrophilic phase further comprises at least one surfactant. 2. The two-phase oral care whitening composition of claim 1, wherein the hydrophobic phase further comprises the surfactant. 3. The two-phase oral care whitening composition of claim 1, wherein the hydrophilic phase further comprises the surfactant. 4. The two-phase oral care whitening composition of claim 1, wherein the hydrophobic phase further comprises a first surfactant, and wherein the hydrophobic phase further comprises a second surfactant. 5. The two-phase oral care whitening composition of claim 4, wherein the first surfactant is a nonionic surfactant. 6. The two-phase oral care whitening composition of claim 4, wherein the second surfactant is an amphoteric surfactant. 7. The two-phase oral care whitening composition of claim 4, wherein the first surfactant is a sorbitan ester. 8. The two-phase oral care whitening composition of claim 4, wherein the second surfactant is cocamidopropyl betaine. 9. The two-phase oral care whitening composition of claim 1, wherein the acyl donor is triacetin. 10. The two-phase oral care whitening composition of claim 1, wherein the hydrophilic phase further comprises a thickener. 11. The two-phase oral care whitening composition of claim 10, wherein the thickener is a carboxypolymethylene. 12. The two-phase oral care whitening composition of claim 1, wherein the source of hydrogen peroxide is a cross-linked polyvinylpyrrolidone (PVP) hydrogen peroxide complex. 13. The two-phase oral care whitening composition of claim 1, wherein the enzyme has perhydrolytic activity and is configured to generate peracetic acid via enzyme-catalyzed perhydrolysis. 14. The two-phase oral care whitening composition of claim 1, wherein the enzyme comprises a CE-7 signature motif that aligns with SEQ ID NO: 2, the CE-7 signature motif comprising a) an RGQ motif at positions corresponding to positions 118-120 of SEQ ID NO: 2; b) a GXSQG (SEQ ID NO: 3) motif at positions corresponding to positions 179-183 of SEQ ID NO: 2 and an HE motif at positions corresponding to positions 298-299 of SEQ ID NO:2. 15. The two-phase oral care whitening composition of claim 1, wherein the enzyme comprises an amino acid sequence comprising a CE-7 signature motif and having at least 80% amino acid sequence identity to SEQ ID NO: 1. 16. The two-phase oral care whitening composition of claim 1, wherein the enzyme comprises SEQ ID NO: 1. 17. A method for whitening teeth, comprising:
contacting the hydrophobic phase and the hydrophilic phase of the two-phase oral care whitening composition of claim 1 with one another to form a mixture; and generating peracetic acid from the mixture. 18. The method of claim 17, further comprising contacting surfaces of the teeth with the peracetic acid generated from the mixture. 19. The method of claim 17, wherein contacting the surfaces of the teeth with the peracetic acid generated from the mixture comprises disposing the mixture in a dental tray. 20. The method of claim 19, wherein contacting the surfaces of the teeth with the peracetic acid generated from the mixture further comprises disposing the dental tray about the teeth to contact the peracetic acid with the surfaces of the teeth. | A two-phase oral care whitening composition including a hydrophobic phase and a hydrophilic phase, and methods for oral care whitening teeth with the same are provided. The hydrophobic phase may include a source of hydrogen peroxide and an acyl donor. The hydrophilic phase may include an enzyme that catalyzes the generation of peracetic acid between the source of hydrogen peroxide and the acyl donor. At least one of the hydrophobic phase and the hydrophilic phase may further include at least one surfactant.1. A two-phase oral care whitening composition, comprising:
a hydrophobic phase comprising a source of hydrogen peroxide and an acyl donor; and a hydrophilic phase comprising an enzyme that catalyzes the generation of peracetic acid between the source of hydrogen peroxide and the acyl donor, wherein at least one of the hydrophobic phase and the hydrophilic phase further comprises at least one surfactant. 2. The two-phase oral care whitening composition of claim 1, wherein the hydrophobic phase further comprises the surfactant. 3. The two-phase oral care whitening composition of claim 1, wherein the hydrophilic phase further comprises the surfactant. 4. The two-phase oral care whitening composition of claim 1, wherein the hydrophobic phase further comprises a first surfactant, and wherein the hydrophobic phase further comprises a second surfactant. 5. The two-phase oral care whitening composition of claim 4, wherein the first surfactant is a nonionic surfactant. 6. The two-phase oral care whitening composition of claim 4, wherein the second surfactant is an amphoteric surfactant. 7. The two-phase oral care whitening composition of claim 4, wherein the first surfactant is a sorbitan ester. 8. The two-phase oral care whitening composition of claim 4, wherein the second surfactant is cocamidopropyl betaine. 9. The two-phase oral care whitening composition of claim 1, wherein the acyl donor is triacetin. 10. The two-phase oral care whitening composition of claim 1, wherein the hydrophilic phase further comprises a thickener. 11. The two-phase oral care whitening composition of claim 10, wherein the thickener is a carboxypolymethylene. 12. The two-phase oral care whitening composition of claim 1, wherein the source of hydrogen peroxide is a cross-linked polyvinylpyrrolidone (PVP) hydrogen peroxide complex. 13. The two-phase oral care whitening composition of claim 1, wherein the enzyme has perhydrolytic activity and is configured to generate peracetic acid via enzyme-catalyzed perhydrolysis. 14. The two-phase oral care whitening composition of claim 1, wherein the enzyme comprises a CE-7 signature motif that aligns with SEQ ID NO: 2, the CE-7 signature motif comprising a) an RGQ motif at positions corresponding to positions 118-120 of SEQ ID NO: 2; b) a GXSQG (SEQ ID NO: 3) motif at positions corresponding to positions 179-183 of SEQ ID NO: 2 and an HE motif at positions corresponding to positions 298-299 of SEQ ID NO:2. 15. The two-phase oral care whitening composition of claim 1, wherein the enzyme comprises an amino acid sequence comprising a CE-7 signature motif and having at least 80% amino acid sequence identity to SEQ ID NO: 1. 16. The two-phase oral care whitening composition of claim 1, wherein the enzyme comprises SEQ ID NO: 1. 17. A method for whitening teeth, comprising:
contacting the hydrophobic phase and the hydrophilic phase of the two-phase oral care whitening composition of claim 1 with one another to form a mixture; and generating peracetic acid from the mixture. 18. The method of claim 17, further comprising contacting surfaces of the teeth with the peracetic acid generated from the mixture. 19. The method of claim 17, wherein contacting the surfaces of the teeth with the peracetic acid generated from the mixture comprises disposing the mixture in a dental tray. 20. The method of claim 19, wherein contacting the surfaces of the teeth with the peracetic acid generated from the mixture further comprises disposing the dental tray about the teeth to contact the peracetic acid with the surfaces of the teeth. | 1,600 |
1,080 | 16,384,782 | 1,632 | The present invention relates to processes for transfecting cells. In particular, the present invention relates to processes for using CRISPR to incorporate a polynucleotide into the genome of an avian primordial germ cell (PGC). | 1. (canceled) 2. A process for using CRISPR to incorporate a polynucleotide into the genome of an avian primordial germ cell (PGC), comprising:
(i) injecting, into a blood vessel of an avian embryo at stage 13-17 that is contained in an egg, a transfection mixture comprising:
(a) a DNA polynucleotide encoding a Type II CRISPR locus comprising a pre-crRNA array, tracrRNA, and a Cas9 nuclease;
(b) a DNA polynucleotide for integration into the genome of one or more germ cells in the avian embryo; and
(c) a cationic lipid,
whereby the DNA polynucleotides enter one or more PGCs in the avian embryo, and
(1) the pre-crRNA array and the tracrRNA of the Type II CRISPR locus are transcribed;
(2) tracrRNA hybridizes to repeat regions of the pre-crRNA and mediates processing of pre-crRNA into mature crRNAs containing individual spacer sequences;
(3) the Cas9 nuclease is expressed and the mature crRNA:tracrRNA complex directs the Cas9 nuclease to the target DNA via Watson-Crick base-pairing between the spacer on the crRNA and a protospacer on the target DNA next to a protospacer adjacent motif (PAM);
(4) the Cas9 nuclease mediates cleavage of target DNA to create a double-stranded break within the protospacer; and
(5) the DNA polynucleotide of (b) is incorporated into the genome of the one or more PGCs in the avian embryo. 3. The process of claim 2, wherein
(i) the cationic lipid is a monovalent cationic lipid selected from one or more of DOTMA (N-[1-(2.3-dioleoyloxy)-propyl]-N,N,N-trimethyl ammonium chloride), DOTAP (1,2-bis(oleoyloxy)-3-3-(trimethylammonium)propane), DMRIE (1,2-dimyristyloxypropyl-3-dimethylhydroxy ethyl ammonium bromide) and DDAB (dimethyl dioctadecyl ammonium bromide), and/or (ii) the cationic lipid is a polyvalent cationic lipid selected from one or more of DOSPA (2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminium trifluoroacetate) and DOSPER (1,3-dioleoyloxy-2-(6carboxy spermyl)-propyl-amid, TMTPS (tetramethyltetrapalmitoyl spermine), TMTOS (tetramethyltetraoleyl spermine), TMTPS (tetramethlytetralauryl spermine), TMTMS (tetramethyltetramyristyl spermine) and TMDOS (tetramethyldioleyl spermine). 4. The process of claim 2, wherein the transfection mixture further comprises a neutral lipid. 5. The process of claim 2, wherein the DNA polynucleotide (b) encodes an RNA molecule with a double-stranded region, or encodes a polypeptide. 6. The process of claim 2, wherein the DNA polynucleotide (b) encodes an siRNA, shRNA or RNA decoy that reduces replication of a virus in a cell, or the DNA polynucleotide encodes an antiviral peptide that reduces replication of a virus in a cell. 7. The process of claim 2, wherein the avian embryo is a stage 13-14 avian embryo. 8. The process of claim 2, wherein the avian is selected from a chicken, duck, turkey, goose, bantam or quail. 9. The process of claim 2, wherein the avian is a chicken. 10. A process for using CRISPR to incorporate a polynucleotide into the genome of an avian primordial germ cell (PGC), comprising:
(i) injecting, into a blood vessel of an avian embryo at stage 13-17 that is contained in an egg, a transfection mixture comprising:
(a) a DNA polynucleotide encoding a chimeric crRNA-tracrRNA hybrid and a Cas9 nuclease;
(b) a DNA polynucleotide for integration into the genome of one or more germ cells in the avian embryo; and
(c) a cationic lipid,
whereby the DNA polynucleotides enter one or more PGCs in the avian embryo, and
(1) the chimeric crRNA-tracrRNA hybrid is transcribed;
(2) the Cas9 nuclease is expressed the chimeric crRNA-tracrRNA hybrid directs the Cas9 nuclease to the target DNA via Watson-Crick base-pairing between the spacer on the chimeric crRNA-tracrRNA hybrid and a protospacer on the target DNA next to a protospacer adjacent motif (PAM);
(4) the Cas9 nuclease mediates cleavage of target DNA to create a double-stranded break within the protospacer; and
(5) the DNA polynucleotide of (b) is incorporated into the genome of the one or more PGCs in the avian embryo. 11. The process of claim 10, wherein
(i) the cationic lipid is a monovalent cationic lipid selected from one or more of DOTMA (N-[1-(2.3-dioleoyloxy)-propyl]-N,N,N-trimethyl ammonium chloride), DOTAP (1,2-bis(oleoyloxy)-3-3-(trimethylammonium)propane), DMRIE (1,2-dimyristyloxypropyl-3-dimethylhydroxy ethyl ammonium bromide) and DDAB (dimethyl dioctadecyl ammonium bromide), and/or (ii) the cationic lipid is a polyvalent cationic lipid selected from one or more of DOSPA (2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminium trifluoroacetate) and DOSPER (1,3-dioleoyloxy-2-(6carboxy spermyl)-propyl-amid, TMTPS (tetramethyltetrapalmitoyl spermine), TMTOS (tetramethyltetraoleyl spermine), TMTPS (tetramethlytetralauryl spermine), TMTMS (tetramethyltetramyristyl spermine) and TMDOS (tetramethyldioleyl spermine). 12. The process of claim 10, wherein the transfection mixture further comprises a neutral lipid. 13. The process of claim 10, wherein the DNA polynucleotide (b) encodes an RNA molecule with a double-stranded region, or encodes a polypeptide. 14. The process of claim 10, wherein the DNA polynucleotide (b) encodes an siRNA, shRNA or RNA decoy that reduces replication of a virus in a cell, or the DNA polynucleotide encodes an antiviral peptide that reduces replication of a virus in a cell. 15. The process of claim 10, wherein the avian embryo is a stage 13-14 avian embryo. 16. The process of claim 10, wherein the avian is selected from a chicken, duck, turkey, goose, bantam or quail. 17. The process of claim 10, wherein the avian is a chicken. | The present invention relates to processes for transfecting cells. In particular, the present invention relates to processes for using CRISPR to incorporate a polynucleotide into the genome of an avian primordial germ cell (PGC).1. (canceled) 2. A process for using CRISPR to incorporate a polynucleotide into the genome of an avian primordial germ cell (PGC), comprising:
(i) injecting, into a blood vessel of an avian embryo at stage 13-17 that is contained in an egg, a transfection mixture comprising:
(a) a DNA polynucleotide encoding a Type II CRISPR locus comprising a pre-crRNA array, tracrRNA, and a Cas9 nuclease;
(b) a DNA polynucleotide for integration into the genome of one or more germ cells in the avian embryo; and
(c) a cationic lipid,
whereby the DNA polynucleotides enter one or more PGCs in the avian embryo, and
(1) the pre-crRNA array and the tracrRNA of the Type II CRISPR locus are transcribed;
(2) tracrRNA hybridizes to repeat regions of the pre-crRNA and mediates processing of pre-crRNA into mature crRNAs containing individual spacer sequences;
(3) the Cas9 nuclease is expressed and the mature crRNA:tracrRNA complex directs the Cas9 nuclease to the target DNA via Watson-Crick base-pairing between the spacer on the crRNA and a protospacer on the target DNA next to a protospacer adjacent motif (PAM);
(4) the Cas9 nuclease mediates cleavage of target DNA to create a double-stranded break within the protospacer; and
(5) the DNA polynucleotide of (b) is incorporated into the genome of the one or more PGCs in the avian embryo. 3. The process of claim 2, wherein
(i) the cationic lipid is a monovalent cationic lipid selected from one or more of DOTMA (N-[1-(2.3-dioleoyloxy)-propyl]-N,N,N-trimethyl ammonium chloride), DOTAP (1,2-bis(oleoyloxy)-3-3-(trimethylammonium)propane), DMRIE (1,2-dimyristyloxypropyl-3-dimethylhydroxy ethyl ammonium bromide) and DDAB (dimethyl dioctadecyl ammonium bromide), and/or (ii) the cationic lipid is a polyvalent cationic lipid selected from one or more of DOSPA (2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminium trifluoroacetate) and DOSPER (1,3-dioleoyloxy-2-(6carboxy spermyl)-propyl-amid, TMTPS (tetramethyltetrapalmitoyl spermine), TMTOS (tetramethyltetraoleyl spermine), TMTPS (tetramethlytetralauryl spermine), TMTMS (tetramethyltetramyristyl spermine) and TMDOS (tetramethyldioleyl spermine). 4. The process of claim 2, wherein the transfection mixture further comprises a neutral lipid. 5. The process of claim 2, wherein the DNA polynucleotide (b) encodes an RNA molecule with a double-stranded region, or encodes a polypeptide. 6. The process of claim 2, wherein the DNA polynucleotide (b) encodes an siRNA, shRNA or RNA decoy that reduces replication of a virus in a cell, or the DNA polynucleotide encodes an antiviral peptide that reduces replication of a virus in a cell. 7. The process of claim 2, wherein the avian embryo is a stage 13-14 avian embryo. 8. The process of claim 2, wherein the avian is selected from a chicken, duck, turkey, goose, bantam or quail. 9. The process of claim 2, wherein the avian is a chicken. 10. A process for using CRISPR to incorporate a polynucleotide into the genome of an avian primordial germ cell (PGC), comprising:
(i) injecting, into a blood vessel of an avian embryo at stage 13-17 that is contained in an egg, a transfection mixture comprising:
(a) a DNA polynucleotide encoding a chimeric crRNA-tracrRNA hybrid and a Cas9 nuclease;
(b) a DNA polynucleotide for integration into the genome of one or more germ cells in the avian embryo; and
(c) a cationic lipid,
whereby the DNA polynucleotides enter one or more PGCs in the avian embryo, and
(1) the chimeric crRNA-tracrRNA hybrid is transcribed;
(2) the Cas9 nuclease is expressed the chimeric crRNA-tracrRNA hybrid directs the Cas9 nuclease to the target DNA via Watson-Crick base-pairing between the spacer on the chimeric crRNA-tracrRNA hybrid and a protospacer on the target DNA next to a protospacer adjacent motif (PAM);
(4) the Cas9 nuclease mediates cleavage of target DNA to create a double-stranded break within the protospacer; and
(5) the DNA polynucleotide of (b) is incorporated into the genome of the one or more PGCs in the avian embryo. 11. The process of claim 10, wherein
(i) the cationic lipid is a monovalent cationic lipid selected from one or more of DOTMA (N-[1-(2.3-dioleoyloxy)-propyl]-N,N,N-trimethyl ammonium chloride), DOTAP (1,2-bis(oleoyloxy)-3-3-(trimethylammonium)propane), DMRIE (1,2-dimyristyloxypropyl-3-dimethylhydroxy ethyl ammonium bromide) and DDAB (dimethyl dioctadecyl ammonium bromide), and/or (ii) the cationic lipid is a polyvalent cationic lipid selected from one or more of DOSPA (2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanaminium trifluoroacetate) and DOSPER (1,3-dioleoyloxy-2-(6carboxy spermyl)-propyl-amid, TMTPS (tetramethyltetrapalmitoyl spermine), TMTOS (tetramethyltetraoleyl spermine), TMTPS (tetramethlytetralauryl spermine), TMTMS (tetramethyltetramyristyl spermine) and TMDOS (tetramethyldioleyl spermine). 12. The process of claim 10, wherein the transfection mixture further comprises a neutral lipid. 13. The process of claim 10, wherein the DNA polynucleotide (b) encodes an RNA molecule with a double-stranded region, or encodes a polypeptide. 14. The process of claim 10, wherein the DNA polynucleotide (b) encodes an siRNA, shRNA or RNA decoy that reduces replication of a virus in a cell, or the DNA polynucleotide encodes an antiviral peptide that reduces replication of a virus in a cell. 15. The process of claim 10, wherein the avian embryo is a stage 13-14 avian embryo. 16. The process of claim 10, wherein the avian is selected from a chicken, duck, turkey, goose, bantam or quail. 17. The process of claim 10, wherein the avian is a chicken. | 1,600 |
1,081 | 15,195,748 | 1,633 | The invention disclosed herein provides systems and methods designed to facilitate human limbal stem/progenitor cell culture including a novel 3-dimensional (3D) sandwich method/system in which human limbal stem/progenitor cells and feeder cells are separately cultured on opposite sides of a porous membrane. | 1. A method of maintaining human limbal stem cells of the corneal epithelium in an undifferentiated human limbal stem cell phenotype, wherein the phenotype is characterized by a small, uniform and compact cellular morpology, the method comprising culturing the human limbal stem cell cells in the system comprising:
a container comprising a culture media for the human limbal stem cells; a porous membrane disposed in the culture media so as to form a first cell culture compartment and a second cell culture compartment, wherein the porous membrane allows soluble factors to migrate between the first cell culture compartment and the second cell culture compartment while simultaneously preventing cells from migrating between the first cell culture compartment and the second cell culture compartment; human limbal stem cells disposed in the first cell culture compartment; human feeder cells disposed in the second cell culture compartment, wherein the human feeder cells secrete one or more soluble factors that faciliate the growth of the human limbal stem cells. 2. The method of claim 1, wherein human limbal stem cell phenotype is characterized by observing expression of at least one of: ATP-binding cassette subfamily G member 2 (ABCG2), Δp63α, or stage-specific embryonic antigen-4 (SSEA4) in the human limbal stem cells. 3. The method of claim 1, wherein the method results in a rate of human limbal stem cells proliferation that is greater that a comparable rate of human limbal stem cell proliferation that is observed in methods that do not use the porous membrane. 4. The method of claim 1, wherein the feeder cells comprise at least one of: human bone marrow derived mesenchymal stem cells, human adipose derived mesenchymal stem cells, or human limbal fibroblasts. 5. The method of claim 1, wherein the system further comprises a sheet of a fibrin material disposed in the container. 6. The method of claim 1, wherein the porous membrane:
comprises a polyethylene terephthalate; and/or comprises pores having a size less than 3 μm. 7. The method of claim 1, wherein the porous membrane is disposed is the system in a horizontal orientation. 8. The method of claim 1, wherein the first cell culture compartment and the second cell culture compartment are arranged in the system so that the mammalian feeder cells are below the human limbal stem cells. 9. The method of claim 1, wherein human limbal stem cells are:
scraped from limbal tissue; and/or pretreated with a protease; and/or pipetted so as to break cell sheets into clusters of cells;
prior to being placed in the first cell culture compartment. 10. The method of claim 1, wherein the human limbal stem cells grow as cell clusters. 11. The method of claim 1, wherein the human limbal stem cells are disposed in the system at a location and in an orientation selected to influence the polarity of the human limbal stem cells. 12. The method of claim 1, wherein the human feeder cells have been treated with mytomycin C or radiation so as to induce growth arrest. 13. The method of claim 1, wherein the feeder cells comprise human bone marrow derived mesenchymal stem cells. 14. The method of claim 1, wherein the feeder cells comprise human adipose derived mesenchymal stem cells. 15. The method of claim 1, wherein the feeder cells comprise human limbal fibroblasts. | The invention disclosed herein provides systems and methods designed to facilitate human limbal stem/progenitor cell culture including a novel 3-dimensional (3D) sandwich method/system in which human limbal stem/progenitor cells and feeder cells are separately cultured on opposite sides of a porous membrane.1. A method of maintaining human limbal stem cells of the corneal epithelium in an undifferentiated human limbal stem cell phenotype, wherein the phenotype is characterized by a small, uniform and compact cellular morpology, the method comprising culturing the human limbal stem cell cells in the system comprising:
a container comprising a culture media for the human limbal stem cells; a porous membrane disposed in the culture media so as to form a first cell culture compartment and a second cell culture compartment, wherein the porous membrane allows soluble factors to migrate between the first cell culture compartment and the second cell culture compartment while simultaneously preventing cells from migrating between the first cell culture compartment and the second cell culture compartment; human limbal stem cells disposed in the first cell culture compartment; human feeder cells disposed in the second cell culture compartment, wherein the human feeder cells secrete one or more soluble factors that faciliate the growth of the human limbal stem cells. 2. The method of claim 1, wherein human limbal stem cell phenotype is characterized by observing expression of at least one of: ATP-binding cassette subfamily G member 2 (ABCG2), Δp63α, or stage-specific embryonic antigen-4 (SSEA4) in the human limbal stem cells. 3. The method of claim 1, wherein the method results in a rate of human limbal stem cells proliferation that is greater that a comparable rate of human limbal stem cell proliferation that is observed in methods that do not use the porous membrane. 4. The method of claim 1, wherein the feeder cells comprise at least one of: human bone marrow derived mesenchymal stem cells, human adipose derived mesenchymal stem cells, or human limbal fibroblasts. 5. The method of claim 1, wherein the system further comprises a sheet of a fibrin material disposed in the container. 6. The method of claim 1, wherein the porous membrane:
comprises a polyethylene terephthalate; and/or comprises pores having a size less than 3 μm. 7. The method of claim 1, wherein the porous membrane is disposed is the system in a horizontal orientation. 8. The method of claim 1, wherein the first cell culture compartment and the second cell culture compartment are arranged in the system so that the mammalian feeder cells are below the human limbal stem cells. 9. The method of claim 1, wherein human limbal stem cells are:
scraped from limbal tissue; and/or pretreated with a protease; and/or pipetted so as to break cell sheets into clusters of cells;
prior to being placed in the first cell culture compartment. 10. The method of claim 1, wherein the human limbal stem cells grow as cell clusters. 11. The method of claim 1, wherein the human limbal stem cells are disposed in the system at a location and in an orientation selected to influence the polarity of the human limbal stem cells. 12. The method of claim 1, wherein the human feeder cells have been treated with mytomycin C or radiation so as to induce growth arrest. 13. The method of claim 1, wherein the feeder cells comprise human bone marrow derived mesenchymal stem cells. 14. The method of claim 1, wherein the feeder cells comprise human adipose derived mesenchymal stem cells. 15. The method of claim 1, wherein the feeder cells comprise human limbal fibroblasts. | 1,600 |
1,082 | 15,537,121 | 1,634 | The present invention provides an oligonucleotide probe for single nucleotide polymorphism detection to be used for a target nucleic acid where a single nucleotide polymorphism is present, the oligonucleotide probe comprising a reporter region, an anchor region, and a linker region. The reporter region comprises: an oligonucleotide consisting of a sequence perfectly matching when a nucleotide of the single nucleotide polymorphism is a first nucleotide, and mismatching when the nucleotide of the single nucleotide polymorphism is a nucleotide other than the first nucleotide; and a fluorescent dye quenching when the reporter region hybridize to the target nucleic acid. | 1. An oligonucleotide probe for single nucleotide polymorphism detection to be used for a target nucleic acid where a single nucleotide polymorphism is present, wherein
the target nucleic acid comprises a first target sequence corresponding to a region comprising the single nucleotide polymorphism, and a second target sequence positioned on a 3′ or 5′ side of the first target sequence and corresponding to a region without the single nucleotide polymorphism, the probe comprises a reporter region for detecting the single nucleotide polymorphism, an anchor region, and a linker region, the reporter region comprises an oligonucleotide consisting of a sequence perfectly matching with the first target sequence when a nucleotide of the single nucleotide polymorphism is a first nucleotide, and mismatching with the first target sequence when the nucleotide is a nucleotide other than the first nucleotide; and a fluorescent dye quenching when the first target sequence and the reporter region hybridize, the anchor region comprises an oligonucleotide consisting of a sequence complementary to the second target sequence, and the linker region links the reporter region and the anchor region to each other, and comprises an oligonucleotide consisting of a sequence non-complementary to a sequence between the first target sequence and the second target sequence in the target nucleic acid. 2. The probe according to claim 1, wherein a length of the oligonucleotide of the reporter region is shorter than a length of the oligonucleotide of the anchor region. 3. The probe according to claim 1, wherein the linker region is an oligonucleotide consisting of a sequence not comprising a universal base. 4. The probe according to claim 1, wherein the linker region is an oligonucleotide consisting of only one kind of base selected from adenine, guanine, cytosine and thymine. 5. The probe according to claim 1, wherein the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 6. A method of detecting a single nucleotide polymorphism, comprising:
preparing a mixture by mixing the probe according to claim 1 and a target nucleic acid comprising a single nucleotide polymorphism; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 7. The probe according to claim 2, wherein the linker region is an oligonucleotide consisting of a sequence not comprising a universal base. 8. The probe according to claim 2, wherein the linker region is an oligonucleotide consisting of only one kind of base selected from adenine, guanine, cytosine and thymine. 9. The probe according to claim 3, wherein the linker region is an oligonucleotide consisting of only one kind of base selected from adenine, guanine, cytosine and thymine. 10. The probe according to claim 2, wherein the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 11. The probe according to claim 3, wherein the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 12. The probe according to claim 4, wherein the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 13. A method of detecting a single nucleotide polymorphism detection in a target nucleic acid, comprising:
preparing a mixture by mixing the probe according to claim 2 and a target nucleic acid comprising a single nucleotide polymorphism; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 14. A method for single nucleotide polymorphism detection, comprising:
preparing a mixture by mixing the probe according to claim 3 and a target nucleic acid comprising a single nucleotide polymorphism; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 15. A method for single nucleotide polymorphism detection, comprising:
preparing a mixture by mixing the probe according to claim 4 and a target nucleic acid where a single nucleotide polymorphism is present; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 16. A method for single nucleotide polymorphism detection, comprising:
preparing a mixture by mixing the probe according to claim 5 and a target nucleic acid where a single nucleotide polymorphism is present; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 17. An oligonucleotide probe for single nucleotide polymorphism detection to be used for a target nucleic acid where a single nucleotide polymorphism is present, wherein
the target nucleic acid comprises a first target sequence corresponding to a region comprising the single nucleotide polymorphism, and a second target sequence positioned on a 3′ or 5′ side of the first target sequence and corresponding to a region without the single nucleotide polymorphism, the probe comprises a reporter region for detecting the single nucleotide polymorphism, an anchor region, and a linker region, the reporter region comprises an oligonucleotide consisting of a sequence perfectly matching with the first target sequence when a nucleotide of the single nucleotide polymorphism is a first nucleotide, and mismatching with the first target sequence when the nucleotide is a nucleotide other than the first nucleotide; and a fluorescent dye quenching when the first target sequence and the reporter region hybridize, the anchor region comprises an oligonucleotide consisting of a sequence complementary to the second target sequence, a length of the oligonucleotide of the reporter region is shorter than a length of the oligonucleotide of the anchor region, the linker region links the reporter region and the anchor region to each other, and comprises an oligonucleotide consisting of a sequence non-complementary to a sequence disposed between the first target sequence and the second target sequence in the target nucleic acid, the linker region is an oligonucleotide consisting of a sequence not comprising a universal base, the linker region is an oligonucleotide consisting of only one kind of base selected from adenine, guanine, cytosine and thymine, and the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 18. A method of detecting a single nucleotide polymorphism, comprising:
preparing a mixture by mixing the probe according to claim 17 and a target nucleic acid comprising a single nucleotide polymorphism; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. | The present invention provides an oligonucleotide probe for single nucleotide polymorphism detection to be used for a target nucleic acid where a single nucleotide polymorphism is present, the oligonucleotide probe comprising a reporter region, an anchor region, and a linker region. The reporter region comprises: an oligonucleotide consisting of a sequence perfectly matching when a nucleotide of the single nucleotide polymorphism is a first nucleotide, and mismatching when the nucleotide of the single nucleotide polymorphism is a nucleotide other than the first nucleotide; and a fluorescent dye quenching when the reporter region hybridize to the target nucleic acid.1. An oligonucleotide probe for single nucleotide polymorphism detection to be used for a target nucleic acid where a single nucleotide polymorphism is present, wherein
the target nucleic acid comprises a first target sequence corresponding to a region comprising the single nucleotide polymorphism, and a second target sequence positioned on a 3′ or 5′ side of the first target sequence and corresponding to a region without the single nucleotide polymorphism, the probe comprises a reporter region for detecting the single nucleotide polymorphism, an anchor region, and a linker region, the reporter region comprises an oligonucleotide consisting of a sequence perfectly matching with the first target sequence when a nucleotide of the single nucleotide polymorphism is a first nucleotide, and mismatching with the first target sequence when the nucleotide is a nucleotide other than the first nucleotide; and a fluorescent dye quenching when the first target sequence and the reporter region hybridize, the anchor region comprises an oligonucleotide consisting of a sequence complementary to the second target sequence, and the linker region links the reporter region and the anchor region to each other, and comprises an oligonucleotide consisting of a sequence non-complementary to a sequence between the first target sequence and the second target sequence in the target nucleic acid. 2. The probe according to claim 1, wherein a length of the oligonucleotide of the reporter region is shorter than a length of the oligonucleotide of the anchor region. 3. The probe according to claim 1, wherein the linker region is an oligonucleotide consisting of a sequence not comprising a universal base. 4. The probe according to claim 1, wherein the linker region is an oligonucleotide consisting of only one kind of base selected from adenine, guanine, cytosine and thymine. 5. The probe according to claim 1, wherein the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 6. A method of detecting a single nucleotide polymorphism, comprising:
preparing a mixture by mixing the probe according to claim 1 and a target nucleic acid comprising a single nucleotide polymorphism; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 7. The probe according to claim 2, wherein the linker region is an oligonucleotide consisting of a sequence not comprising a universal base. 8. The probe according to claim 2, wherein the linker region is an oligonucleotide consisting of only one kind of base selected from adenine, guanine, cytosine and thymine. 9. The probe according to claim 3, wherein the linker region is an oligonucleotide consisting of only one kind of base selected from adenine, guanine, cytosine and thymine. 10. The probe according to claim 2, wherein the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 11. The probe according to claim 3, wherein the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 12. The probe according to claim 4, wherein the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 13. A method of detecting a single nucleotide polymorphism detection in a target nucleic acid, comprising:
preparing a mixture by mixing the probe according to claim 2 and a target nucleic acid comprising a single nucleotide polymorphism; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 14. A method for single nucleotide polymorphism detection, comprising:
preparing a mixture by mixing the probe according to claim 3 and a target nucleic acid comprising a single nucleotide polymorphism; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 15. A method for single nucleotide polymorphism detection, comprising:
preparing a mixture by mixing the probe according to claim 4 and a target nucleic acid where a single nucleotide polymorphism is present; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 16. A method for single nucleotide polymorphism detection, comprising:
preparing a mixture by mixing the probe according to claim 5 and a target nucleic acid where a single nucleotide polymorphism is present; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. 17. An oligonucleotide probe for single nucleotide polymorphism detection to be used for a target nucleic acid where a single nucleotide polymorphism is present, wherein
the target nucleic acid comprises a first target sequence corresponding to a region comprising the single nucleotide polymorphism, and a second target sequence positioned on a 3′ or 5′ side of the first target sequence and corresponding to a region without the single nucleotide polymorphism, the probe comprises a reporter region for detecting the single nucleotide polymorphism, an anchor region, and a linker region, the reporter region comprises an oligonucleotide consisting of a sequence perfectly matching with the first target sequence when a nucleotide of the single nucleotide polymorphism is a first nucleotide, and mismatching with the first target sequence when the nucleotide is a nucleotide other than the first nucleotide; and a fluorescent dye quenching when the first target sequence and the reporter region hybridize, the anchor region comprises an oligonucleotide consisting of a sequence complementary to the second target sequence, a length of the oligonucleotide of the reporter region is shorter than a length of the oligonucleotide of the anchor region, the linker region links the reporter region and the anchor region to each other, and comprises an oligonucleotide consisting of a sequence non-complementary to a sequence disposed between the first target sequence and the second target sequence in the target nucleic acid, the linker region is an oligonucleotide consisting of a sequence not comprising a universal base, the linker region is an oligonucleotide consisting of only one kind of base selected from adenine, guanine, cytosine and thymine, and the linker region is an oligonucleotide consisting of 3 to 11 nucleotides. 18. A method of detecting a single nucleotide polymorphism, comprising:
preparing a mixture by mixing the probe according to claim 17 and a target nucleic acid comprising a single nucleotide polymorphism; measuring fluorescence intensity of the mixture; and detecting the presence/absence of the single nucleotide polymorphism in the target nucleic acid on the basis of the fluorescence intensity. | 1,600 |
1,083 | 15,788,255 | 1,612 | Oral care compositions comprising an antifibrinolytic agent (e.g., tranexamic acid) and stannous ion source are provided for promoting Gum Health of a consumer. | 1. An oral care composition comprising:
a) from 0.01% to 5%, by weight of the composition, of a stannous ion source; and b) from 0.01% to 10%, by weight of the composition, of an antifibrinolytic agent. 2. The oral care composition according to claim 1, wherein the antifibrinolytic agent is a compound of formula (I):
H2N—X—COOH (I)
wherein X is a branched or unbranched, saturated or unsaturated, aliphatic group; or an aromatic group. 3. The oral care composition according to claim 1, wherein the antifibrinolytic agent is present in an amount of less than 5%, by weight of the composition, and selected from tranexamic acid, epsilon aminocaproic acid, p-aminomethylbenzoic acid, or combinations thereof. 4. The oral care composition according to claim 3, wherein the antifibrinolytic agent is tranexamic acid, and present in an amount of from 0.1% to less than 1% by weight of the composition. 5. A oral care composition, comprising:
a) from 0.01% to 5%, by weight of the composition, of a stannous ion source; and b) from 0.01% to 10%, by weight of the composition, of one or more compounds selected from the group consisting of tranexamic acid, epsilon aminocaproic acid, p-aminomethylbenzoic acid, and combinations thereof. 6. The oral care composition according claim 1, wherein the stannous ion source is present in the amount of from 0.05% to 4% by weight of the composition, and is selected from the group consisting of stannous chloride, stannous fluoride, stannous acetate, stannous gluconate, stannous oxalate, stannous sulfate, stannous lactate, stannous tartrate, stannous iodide, stannous chlorofluoride, stannous hexafluorozirconate, stannous citrate, stannous malate, stannous glycinate, stannous carbonate, stannous phosphate, stannous pyrophosphate, stannous metaphosphate, and combinations thereof. 7. The oral care composition according to claim 1, comprising from 0.01% to 5%, by weight of the composition, of a thickening agent comprising at least one agent selected from the group consisting of:
(i) a linear sulfated polysaccharide; (ii) a natural gum; (iii) a non-ionic cellulose or derivative thereof; (iv) a polyvinyl pyrrolidone (PVP); (v) a polymer comprising at least a polycarboxylated ethylene backbone; (vi) polyacrylamide; (vii) co-polymers comprising acrylamide; (viii) pectin; (ix) proteins; (x) polyethylene glycols (PEG), preferably high molecular weight PEG; and (xi) combinations thereof. 8. The oral care composition according to claim 7, wherein the thickening agent is present in an amount of from 1% to 2.5%, by weight of the composition, wherein:
(i) the linear sulfated polysaccharide is a carrageenan; (ii) the natural gum is selected from the group consisting of xanthan gum, gum karaya, gum arabic, gum tragacanth, and combinations thereof; (iii) the non-ionic cellulose or derivative thereof having an average molecular weight range of 50,000 to 1,300,000 Daltons, and preferably an average degree of polymerization from 300 to 4,800; (iv) the polymer comprising at least a polycarboxylated ethylene backbone and selected from the group consisting of: co-polymers of maleic anhydride with methyl vinyl ether having a molecular weight of 30,000 to 1,000,000 Daltons; homo-polymers of acrylic acid; and co-polymers of maleic acid and acrylic acid or methacrylic; and (v) combinations thereof. 9. The oral care composition according to claim 8, wherein:
(i) the carrageenan is selected from the group consisting of Kappa-carrageenan, Iota-carrageenan, Lambda-carrageenan, and combinations thereof; (ii) the natural gum is xanthan gum; (iii) the non-ionic cellulose or derivative thereof is hydroxyethyl cellulose (HEC); (iv) the polymer comprising at least a polycarboxylated ethylene backbone is the co-polymers of maleic acid or anhydride with methyl vinyl ether; and (v) combinations thereof. 10. The oral care composition according to claim 1, comprising from 0% to less than 0.001%, by weight of the composition folic acid. 11. The oral care composition according to claim 1 comprising: from 0.01% to 5% by weight of the composition, of a flavor composition, wherein the flavor composition comprises:
(i) a flavor mixture comprising from greater than 0% to less than 55%, or from greater than 65% to 95%, by weight of the flavor composition, of methyl salicyclate; from greater than 0% to less than 30%, or from greater than 35% to 65%, by weight of the flavor composition, of menthol; from greater than 0% to less than 1%, or from greater than 5% to 50%, by weight of the flavor composition, of eugenol; and from greater than 0% to less than 3%, or from greater than 8% to 30%, by weight of the flavor composition, of cineol; or
(ii) a flavor mixture that is free or substantially free of methyl salicyclate, menthol, eugenol, and cineol. 12. The oral care composition according claim 1 comprising from 0.0025% to 2% by weight of the composition, of a fluoride ion source, wherein the fluoride ion source is selected from the group consisting of sodium fluoride, indium fluoride, amine fluoride, sodium monofluorophosphate (MFP), potassium fluoride, zinc fluoride, and combinations thereof. 13. The oral care composition according to claim 1 comprising from 10% to 70% water. 14. The oral care composition according to claim 1, wherein the composition has a pH from 4.5 to 11. 15. A method of promoting Gum Health in a human subject comprising administering to the subject's oral cavity an oral care composition according to any one of claims 1 to 14, preferably once a day, more preferably twice a day. 16. The method of claim 15, wherein the promotion of Gum Health occurs within a period selected from the group consisting of:
a) from time 0 hours to 72 hours; b) from time 0 hours to 48 hours; c) from time 0 hours to 24 hours; wherein time 0 hours is upon the administration of the oral care composition. 17. The method of claim 15 or 16, wherein Gum Health is selected from:
(i) improving gingival wound healing in the oral cavity;
(ii) improving reduction of bacterial activity in the oral cavity; or
(iii) combination thereof. | Oral care compositions comprising an antifibrinolytic agent (e.g., tranexamic acid) and stannous ion source are provided for promoting Gum Health of a consumer.1. An oral care composition comprising:
a) from 0.01% to 5%, by weight of the composition, of a stannous ion source; and b) from 0.01% to 10%, by weight of the composition, of an antifibrinolytic agent. 2. The oral care composition according to claim 1, wherein the antifibrinolytic agent is a compound of formula (I):
H2N—X—COOH (I)
wherein X is a branched or unbranched, saturated or unsaturated, aliphatic group; or an aromatic group. 3. The oral care composition according to claim 1, wherein the antifibrinolytic agent is present in an amount of less than 5%, by weight of the composition, and selected from tranexamic acid, epsilon aminocaproic acid, p-aminomethylbenzoic acid, or combinations thereof. 4. The oral care composition according to claim 3, wherein the antifibrinolytic agent is tranexamic acid, and present in an amount of from 0.1% to less than 1% by weight of the composition. 5. A oral care composition, comprising:
a) from 0.01% to 5%, by weight of the composition, of a stannous ion source; and b) from 0.01% to 10%, by weight of the composition, of one or more compounds selected from the group consisting of tranexamic acid, epsilon aminocaproic acid, p-aminomethylbenzoic acid, and combinations thereof. 6. The oral care composition according claim 1, wherein the stannous ion source is present in the amount of from 0.05% to 4% by weight of the composition, and is selected from the group consisting of stannous chloride, stannous fluoride, stannous acetate, stannous gluconate, stannous oxalate, stannous sulfate, stannous lactate, stannous tartrate, stannous iodide, stannous chlorofluoride, stannous hexafluorozirconate, stannous citrate, stannous malate, stannous glycinate, stannous carbonate, stannous phosphate, stannous pyrophosphate, stannous metaphosphate, and combinations thereof. 7. The oral care composition according to claim 1, comprising from 0.01% to 5%, by weight of the composition, of a thickening agent comprising at least one agent selected from the group consisting of:
(i) a linear sulfated polysaccharide; (ii) a natural gum; (iii) a non-ionic cellulose or derivative thereof; (iv) a polyvinyl pyrrolidone (PVP); (v) a polymer comprising at least a polycarboxylated ethylene backbone; (vi) polyacrylamide; (vii) co-polymers comprising acrylamide; (viii) pectin; (ix) proteins; (x) polyethylene glycols (PEG), preferably high molecular weight PEG; and (xi) combinations thereof. 8. The oral care composition according to claim 7, wherein the thickening agent is present in an amount of from 1% to 2.5%, by weight of the composition, wherein:
(i) the linear sulfated polysaccharide is a carrageenan; (ii) the natural gum is selected from the group consisting of xanthan gum, gum karaya, gum arabic, gum tragacanth, and combinations thereof; (iii) the non-ionic cellulose or derivative thereof having an average molecular weight range of 50,000 to 1,300,000 Daltons, and preferably an average degree of polymerization from 300 to 4,800; (iv) the polymer comprising at least a polycarboxylated ethylene backbone and selected from the group consisting of: co-polymers of maleic anhydride with methyl vinyl ether having a molecular weight of 30,000 to 1,000,000 Daltons; homo-polymers of acrylic acid; and co-polymers of maleic acid and acrylic acid or methacrylic; and (v) combinations thereof. 9. The oral care composition according to claim 8, wherein:
(i) the carrageenan is selected from the group consisting of Kappa-carrageenan, Iota-carrageenan, Lambda-carrageenan, and combinations thereof; (ii) the natural gum is xanthan gum; (iii) the non-ionic cellulose or derivative thereof is hydroxyethyl cellulose (HEC); (iv) the polymer comprising at least a polycarboxylated ethylene backbone is the co-polymers of maleic acid or anhydride with methyl vinyl ether; and (v) combinations thereof. 10. The oral care composition according to claim 1, comprising from 0% to less than 0.001%, by weight of the composition folic acid. 11. The oral care composition according to claim 1 comprising: from 0.01% to 5% by weight of the composition, of a flavor composition, wherein the flavor composition comprises:
(i) a flavor mixture comprising from greater than 0% to less than 55%, or from greater than 65% to 95%, by weight of the flavor composition, of methyl salicyclate; from greater than 0% to less than 30%, or from greater than 35% to 65%, by weight of the flavor composition, of menthol; from greater than 0% to less than 1%, or from greater than 5% to 50%, by weight of the flavor composition, of eugenol; and from greater than 0% to less than 3%, or from greater than 8% to 30%, by weight of the flavor composition, of cineol; or
(ii) a flavor mixture that is free or substantially free of methyl salicyclate, menthol, eugenol, and cineol. 12. The oral care composition according claim 1 comprising from 0.0025% to 2% by weight of the composition, of a fluoride ion source, wherein the fluoride ion source is selected from the group consisting of sodium fluoride, indium fluoride, amine fluoride, sodium monofluorophosphate (MFP), potassium fluoride, zinc fluoride, and combinations thereof. 13. The oral care composition according to claim 1 comprising from 10% to 70% water. 14. The oral care composition according to claim 1, wherein the composition has a pH from 4.5 to 11. 15. A method of promoting Gum Health in a human subject comprising administering to the subject's oral cavity an oral care composition according to any one of claims 1 to 14, preferably once a day, more preferably twice a day. 16. The method of claim 15, wherein the promotion of Gum Health occurs within a period selected from the group consisting of:
a) from time 0 hours to 72 hours; b) from time 0 hours to 48 hours; c) from time 0 hours to 24 hours; wherein time 0 hours is upon the administration of the oral care composition. 17. The method of claim 15 or 16, wherein Gum Health is selected from:
(i) improving gingival wound healing in the oral cavity;
(ii) improving reduction of bacterial activity in the oral cavity; or
(iii) combination thereof. | 1,600 |
1,084 | 15,973,243 | 1,637 | Digital assay system, including methods, apparatus, and compositions, for assay of one or more targets in a set of partitions containing a generic reporter of target amplification. | 1. A method of performing a digital assay, the method comprising:
forming partitions each including a portion of a same mixture, the mixture containing a target and also containing a generic reporter that is sensitive to amplification of the target, wherein only a subset of the partitions each contain at least one copy of the target; amplifying the target and at least one byproduct in the partitions; collecting amplification data from the generic reporter for a plurality of the partitions, wherein partitions exhibiting detectable amplification of the target, partitions exhibiting detectable amplification of the byproduct, and partitions exhibiting detectable amplification of neither the target nor the byproduct are distinguishable from one another in the data; and determining a level of the target using the amplification data. 2. The method of claim 1, wherein the byproduct includes a primer dimer. 3. The method of claim 1, wherein detectable amplification of the byproduct occurs stochastically in the plurality of partitions. 4. The method of claim 1, wherein detectable amplification of the target in a partition inhibits detectable amplification of the byproduct. 5. The method of claim 1, further comprising a step of assigning partitions of the plurality as positive or negative for the target based on a threshold that selectively includes partitions exhibiting detectable amplification of the target and selectively excludes partitions exhibiting detectable amplification of the byproduct. 6. The method of claim 5, wherein the step of assigning includes a step of comparing a signal detected from each partition of the plurality with the threshold, and wherein the signal represents photoluminescence detected from the generic reporter. 7. The method of claim 5, wherein the threshold corresponds to a strength of the signal that is intermediate respective signal strengths for partitions exhibiting detectable amplification of the target and partitions exhibiting detectable amplification of the byproduct. 8. The method of claim 5, wherein partitions exhibiting detectable amplification of neither the target nor the byproduct have a signal strength that is less than the signal strength of partitions exhibiting detectable amplification of the byproduct. 9. The method of claim 5, further comprising a step of choosing the threshold based on respective signal strengths of partitions exhibiting detectable amplification of the target and partitions exhibiting detectable amplification of the byproduct. 10. The method of claim 5, wherein partitions exhibiting detectable amplification of the target have a stronger signal than partitions exhibiting detectable amplification of the byproduct. 11. The method of claim 5, wherein the step of determining a level of the target includes a step of calculating a concentration based on a number of the plurality of partitions that are assigned as positive for the target or based on a number of the plurality of partitions that are assigned as negative for the target. | Digital assay system, including methods, apparatus, and compositions, for assay of one or more targets in a set of partitions containing a generic reporter of target amplification.1. A method of performing a digital assay, the method comprising:
forming partitions each including a portion of a same mixture, the mixture containing a target and also containing a generic reporter that is sensitive to amplification of the target, wherein only a subset of the partitions each contain at least one copy of the target; amplifying the target and at least one byproduct in the partitions; collecting amplification data from the generic reporter for a plurality of the partitions, wherein partitions exhibiting detectable amplification of the target, partitions exhibiting detectable amplification of the byproduct, and partitions exhibiting detectable amplification of neither the target nor the byproduct are distinguishable from one another in the data; and determining a level of the target using the amplification data. 2. The method of claim 1, wherein the byproduct includes a primer dimer. 3. The method of claim 1, wherein detectable amplification of the byproduct occurs stochastically in the plurality of partitions. 4. The method of claim 1, wherein detectable amplification of the target in a partition inhibits detectable amplification of the byproduct. 5. The method of claim 1, further comprising a step of assigning partitions of the plurality as positive or negative for the target based on a threshold that selectively includes partitions exhibiting detectable amplification of the target and selectively excludes partitions exhibiting detectable amplification of the byproduct. 6. The method of claim 5, wherein the step of assigning includes a step of comparing a signal detected from each partition of the plurality with the threshold, and wherein the signal represents photoluminescence detected from the generic reporter. 7. The method of claim 5, wherein the threshold corresponds to a strength of the signal that is intermediate respective signal strengths for partitions exhibiting detectable amplification of the target and partitions exhibiting detectable amplification of the byproduct. 8. The method of claim 5, wherein partitions exhibiting detectable amplification of neither the target nor the byproduct have a signal strength that is less than the signal strength of partitions exhibiting detectable amplification of the byproduct. 9. The method of claim 5, further comprising a step of choosing the threshold based on respective signal strengths of partitions exhibiting detectable amplification of the target and partitions exhibiting detectable amplification of the byproduct. 10. The method of claim 5, wherein partitions exhibiting detectable amplification of the target have a stronger signal than partitions exhibiting detectable amplification of the byproduct. 11. The method of claim 5, wherein the step of determining a level of the target includes a step of calculating a concentration based on a number of the plurality of partitions that are assigned as positive for the target or based on a number of the plurality of partitions that are assigned as negative for the target. | 1,600 |
1,085 | 15,553,638 | 1,622 | Described are also corresponding pharmaceutical formulations, cosmetic preparations and preparations for pleasure and/or nutrition, suitable for consumption, as well as methods for producing CBDV and THCV. | 1. A mixture comprising one or more compounds of formula (A) and/or one or more of their salts
wherein R1 is an aliphatic rest with none or one, two, three or more than three hydroxyl groups, wherein the total number of C-atoms in the aliphatic rest R1 is not bigger than 15,
an wherein the aliphatic rest is
saturated or unsaturated,
branched or linear, and
acyclic or cyclic,
provided that if R1 is an aliphatic rest with no hydroxyl group, the compound(s) of the formula (A) or, respectively, the salt(s) thereof, is/are selected from the group consisting of the compounds cyclohexylcannabidivarinolat and hexylcannabidivarinolat as well as their salts,
wherein in the mixture the molar ratio of the total amount of compounds of the formula (A) and their salts to the amount of compounds of formula (V) (cannabidivarin, CBDV)
is bigger than 1:1 and simultaneously the molar ratio of the total amount of compounds of the formula (A) and their salts to the amount of compounds of formula (III) ((−)-trans-methylcannabidivarinolat)
is bigger than 1:1. 2. The mixture according to claim 1, wherein the aliphatic rest of the compound of formula (A) is saturated and/or linear. 3. The mixture according to claim 1, wherein the compound of the formula (A) is a compound of formula (A-I)
wherein applies:
each R1, independently of the meaning of each of the others of the overall n rests R1 is H, alkyl with one or two C-atoms or OH,
R2 is H or OH,
n is an integral number in the range from 2 to 10,
wherein at least one of the rests R1 or the rest R2 is OH,
and/or a compound of the formula ) (A-II)
wherein applies:
each R1 independently of the meaning of each of the others of the overall n−1 rests R1 is H, alkyl with one or two C-atoms or OH,
R2 is H or OH,
n is an integral number in the range from 2 to 10,
wherein at least one of the rests R1 or the rest R2 is OH. 4. A mixture according to claim 1, wherein the compound of the formula (A) is
(i) a compound of formula (A-III)
wherein applies:
each R1 independently of the meaning of each of the others of the overall n−2 rests R1 is H, alkyl with one or two C-atoms or OH,
R2 is H or OH,
n is an integral number in the range from 2 to 10,
wherein at least one of the rests R1 or the rest R2 is OH, and/or
(ii) a compound of formula (A-IV)
wherein applies:
each R1 independently of the meaning of each of the others of the overall n−1 rests R1 is H, alkyl with one or two C-atoms or OH, and
n is an integral number in the range from 2 to 10. 5. A mixture according to claim 3, wherein in the said formulas (A-I), (A-II), (A-III) or, respectively, (A-IV) each R1 independently of the meaning of each of the others of the rests R1 is H or OH. 6. A mixture according to claim 1, wherein the compound of the formula (A) is a compound of formula (TV): 7. A method of therapeutic treatment of a human or animal comprising administering to the human or animal a mixture of claim 1, or a compound of the formula (A), or salt thereof. 8. A method for achieving an effect selected from the group consisting of:
appetizing effect, antiemetic effect for the inhibition of nausea and vomiting, reduction of muscular cramps and spasticities, alleviation of pain symptoms, alleviation of migraine symptoms, reduction of the intraocular pressure in the case of a glaucoma, improved sentiment, immune stimulation, antiepileptic effect, and as CB1 and/or CB2 receptor modulator; the method comprising administering to the human or animal a mixture of claim 1 or a compound of the formula (A), or salt thereof. 9. A method for producing a mixture according to claim 1 or a compound selected from the group consisting of cyclohexylcannabidivarinolat, hexylcannabidivarinolat and 2-hydroxyethylcannabidivarinolat or a salt thereof, comprising the following step:
reacting an ester of the formula (IX)
wherein Y is an organic rest,
with an alcohol of the formula HO—X,
wherein X is an aliphatic rest with none, or one, two, three or more than three hydroxyl groups,
wherein the total amount of C-atoms in the aliphatic rest X is not bigger than 15, and
wherein the aliphatic rest is
saturated or unsaturated,
branched or linear, and
acyclic or cyclic,
provided that if X is an aliphatic rest with no hydroxyl group, the alcohol of the formula HO—X is selected from the group consisting of cyclohexanol and hexanol,
wherein Y is different from X and selected in a way that an alcohol of the formula HO—Y formed during the reaction boils at 1013 hPa at a lower temperature than the used alcohol of the formula HO—X. 10. The method according to claim 9 comprising the following step to produce the ester of formula (III):
reacting menthadienol of the formula (I) with a divarin ester of the formula (II) to the corresponding ester of formula (III), 11. A method for producing a mixture according to claim 1 or a compound selected from the group consisting of cyclohexylcannabidivarinolat, hexylcannabidivarinolat and 2-hydroxyethylcannabidivarinolat or a salt thereof, comprising the following steps:
(a) transesterification of a divarinacid ester,
and
(b) reacting the divarinacid ester obtained by transesterification in step (a) with menthadienol to the corresponding compound of the formula (A). 12. A method for producing THCV, comprising the production of a mixture according to claim 1 or of a compound selected from the group consisting of cyclohexylcannabidivarinolat, hexylcannabidivarinolat and 2-hydroxyethylcannabidivarinolat or a salt thereof. 13. A method according to claim 12, wherein the produced mixture or, respectively, the compound of the formula (A) or the salt thereof is treated in a way, that the compound of the formula (A) present in the mixture or, respectively, the salt thereof is decarboxylatingly saponified and the compound of the formula (V) (cannabidivarin, CBDV)
is formed. 14. A compound selected from the group consisting of cyclohexylcannabidivarinolat, hexylcannabidivarinolat, and 2-hydroxyethylcannabidivarinolat. 15. A pharmaceutical formulation or cosmetic preparation, comprising a mixture according to claim 1. 16. A preparation for nutrition and/or pleasure, suitable for consumption, comprising a mixture according to claim 1. 17. A pharmaceutical formulation or cosmetic preparation, comprising one or more compounds according to claim 14 and/or one or more salts thereof. 18. A preparation for nutrition and/or pleasure, suitable for consumption, comprising one or more compounds according to claim 14 and/or one or more salts thereof. 19. Crystalline cannabidivarin of formula (V) | Described are also corresponding pharmaceutical formulations, cosmetic preparations and preparations for pleasure and/or nutrition, suitable for consumption, as well as methods for producing CBDV and THCV.1. A mixture comprising one or more compounds of formula (A) and/or one or more of their salts
wherein R1 is an aliphatic rest with none or one, two, three or more than three hydroxyl groups, wherein the total number of C-atoms in the aliphatic rest R1 is not bigger than 15,
an wherein the aliphatic rest is
saturated or unsaturated,
branched or linear, and
acyclic or cyclic,
provided that if R1 is an aliphatic rest with no hydroxyl group, the compound(s) of the formula (A) or, respectively, the salt(s) thereof, is/are selected from the group consisting of the compounds cyclohexylcannabidivarinolat and hexylcannabidivarinolat as well as their salts,
wherein in the mixture the molar ratio of the total amount of compounds of the formula (A) and their salts to the amount of compounds of formula (V) (cannabidivarin, CBDV)
is bigger than 1:1 and simultaneously the molar ratio of the total amount of compounds of the formula (A) and their salts to the amount of compounds of formula (III) ((−)-trans-methylcannabidivarinolat)
is bigger than 1:1. 2. The mixture according to claim 1, wherein the aliphatic rest of the compound of formula (A) is saturated and/or linear. 3. The mixture according to claim 1, wherein the compound of the formula (A) is a compound of formula (A-I)
wherein applies:
each R1, independently of the meaning of each of the others of the overall n rests R1 is H, alkyl with one or two C-atoms or OH,
R2 is H or OH,
n is an integral number in the range from 2 to 10,
wherein at least one of the rests R1 or the rest R2 is OH,
and/or a compound of the formula ) (A-II)
wherein applies:
each R1 independently of the meaning of each of the others of the overall n−1 rests R1 is H, alkyl with one or two C-atoms or OH,
R2 is H or OH,
n is an integral number in the range from 2 to 10,
wherein at least one of the rests R1 or the rest R2 is OH. 4. A mixture according to claim 1, wherein the compound of the formula (A) is
(i) a compound of formula (A-III)
wherein applies:
each R1 independently of the meaning of each of the others of the overall n−2 rests R1 is H, alkyl with one or two C-atoms or OH,
R2 is H or OH,
n is an integral number in the range from 2 to 10,
wherein at least one of the rests R1 or the rest R2 is OH, and/or
(ii) a compound of formula (A-IV)
wherein applies:
each R1 independently of the meaning of each of the others of the overall n−1 rests R1 is H, alkyl with one or two C-atoms or OH, and
n is an integral number in the range from 2 to 10. 5. A mixture according to claim 3, wherein in the said formulas (A-I), (A-II), (A-III) or, respectively, (A-IV) each R1 independently of the meaning of each of the others of the rests R1 is H or OH. 6. A mixture according to claim 1, wherein the compound of the formula (A) is a compound of formula (TV): 7. A method of therapeutic treatment of a human or animal comprising administering to the human or animal a mixture of claim 1, or a compound of the formula (A), or salt thereof. 8. A method for achieving an effect selected from the group consisting of:
appetizing effect, antiemetic effect for the inhibition of nausea and vomiting, reduction of muscular cramps and spasticities, alleviation of pain symptoms, alleviation of migraine symptoms, reduction of the intraocular pressure in the case of a glaucoma, improved sentiment, immune stimulation, antiepileptic effect, and as CB1 and/or CB2 receptor modulator; the method comprising administering to the human or animal a mixture of claim 1 or a compound of the formula (A), or salt thereof. 9. A method for producing a mixture according to claim 1 or a compound selected from the group consisting of cyclohexylcannabidivarinolat, hexylcannabidivarinolat and 2-hydroxyethylcannabidivarinolat or a salt thereof, comprising the following step:
reacting an ester of the formula (IX)
wherein Y is an organic rest,
with an alcohol of the formula HO—X,
wherein X is an aliphatic rest with none, or one, two, three or more than three hydroxyl groups,
wherein the total amount of C-atoms in the aliphatic rest X is not bigger than 15, and
wherein the aliphatic rest is
saturated or unsaturated,
branched or linear, and
acyclic or cyclic,
provided that if X is an aliphatic rest with no hydroxyl group, the alcohol of the formula HO—X is selected from the group consisting of cyclohexanol and hexanol,
wherein Y is different from X and selected in a way that an alcohol of the formula HO—Y formed during the reaction boils at 1013 hPa at a lower temperature than the used alcohol of the formula HO—X. 10. The method according to claim 9 comprising the following step to produce the ester of formula (III):
reacting menthadienol of the formula (I) with a divarin ester of the formula (II) to the corresponding ester of formula (III), 11. A method for producing a mixture according to claim 1 or a compound selected from the group consisting of cyclohexylcannabidivarinolat, hexylcannabidivarinolat and 2-hydroxyethylcannabidivarinolat or a salt thereof, comprising the following steps:
(a) transesterification of a divarinacid ester,
and
(b) reacting the divarinacid ester obtained by transesterification in step (a) with menthadienol to the corresponding compound of the formula (A). 12. A method for producing THCV, comprising the production of a mixture according to claim 1 or of a compound selected from the group consisting of cyclohexylcannabidivarinolat, hexylcannabidivarinolat and 2-hydroxyethylcannabidivarinolat or a salt thereof. 13. A method according to claim 12, wherein the produced mixture or, respectively, the compound of the formula (A) or the salt thereof is treated in a way, that the compound of the formula (A) present in the mixture or, respectively, the salt thereof is decarboxylatingly saponified and the compound of the formula (V) (cannabidivarin, CBDV)
is formed. 14. A compound selected from the group consisting of cyclohexylcannabidivarinolat, hexylcannabidivarinolat, and 2-hydroxyethylcannabidivarinolat. 15. A pharmaceutical formulation or cosmetic preparation, comprising a mixture according to claim 1. 16. A preparation for nutrition and/or pleasure, suitable for consumption, comprising a mixture according to claim 1. 17. A pharmaceutical formulation or cosmetic preparation, comprising one or more compounds according to claim 14 and/or one or more salts thereof. 18. A preparation for nutrition and/or pleasure, suitable for consumption, comprising one or more compounds according to claim 14 and/or one or more salts thereof. 19. Crystalline cannabidivarin of formula (V) | 1,600 |
1,086 | 13,675,306 | 1,615 | An improved method for diagnosing and characterizing peripheral nerve lesions to permit early identification and characterization of peripheral nerve injuries that will require surgical intervention. | 1. A method to diagnose and characterize peripheral nerve lesions, the method including the step of:
introducing a chemical agent operable to bind directly to a peripheral nerve lesion in which the blood-nerve barrier is compromised, wherein the chemical agent does not interact with a target molecule unless the blood-nerve barrier is compromised. 2. The method of claim 1, further comprising the step of:
attaching a label to the chemical agent, wherein the chemical agent with the label is operable to localize in regions where a peripheral of the blood-nerve barrier is disrupted. 3. The method of claim 2, further comprising the step of:
quantitatively analyzing the peripheral nerve or nerves to determine the amount of labeling agent present. 4. The method of claim 3,
wherein the analyzing is performed using magnetic resonance imaging techniques. 5. The method of claim 1,
wherein the chemical agent is a monoclonal antibody. 6. The method of claim 1,
wherein the chemical agent is also operable to bind specifically and exclusively to the peripheral nerve lesions in which the blood-nerve barrier is compromised. 7. The method of claim 1,
wherein the peripheral nerve lesions in which the blood-nerve barrier is compromised is an antibody against an axonal protein. 8. The method of claim 1,
wherein the blood-nerve barrier is compromised as in neurotmesis. 9. The method of claim 1,
wherein the label attached to the chemical is gadolinium-DTPA or superparamagnetic iron oxide nanoparticles. 10. The method of claim 1,
wherein the method enables the accurate analysis of peripheral nerve lesions and the ability to distinguish axonotmetic lesions with greater self-regeneration potential from neurotmetic lesions that may require intervention. | An improved method for diagnosing and characterizing peripheral nerve lesions to permit early identification and characterization of peripheral nerve injuries that will require surgical intervention.1. A method to diagnose and characterize peripheral nerve lesions, the method including the step of:
introducing a chemical agent operable to bind directly to a peripheral nerve lesion in which the blood-nerve barrier is compromised, wherein the chemical agent does not interact with a target molecule unless the blood-nerve barrier is compromised. 2. The method of claim 1, further comprising the step of:
attaching a label to the chemical agent, wherein the chemical agent with the label is operable to localize in regions where a peripheral of the blood-nerve barrier is disrupted. 3. The method of claim 2, further comprising the step of:
quantitatively analyzing the peripheral nerve or nerves to determine the amount of labeling agent present. 4. The method of claim 3,
wherein the analyzing is performed using magnetic resonance imaging techniques. 5. The method of claim 1,
wherein the chemical agent is a monoclonal antibody. 6. The method of claim 1,
wherein the chemical agent is also operable to bind specifically and exclusively to the peripheral nerve lesions in which the blood-nerve barrier is compromised. 7. The method of claim 1,
wherein the peripheral nerve lesions in which the blood-nerve barrier is compromised is an antibody against an axonal protein. 8. The method of claim 1,
wherein the blood-nerve barrier is compromised as in neurotmesis. 9. The method of claim 1,
wherein the label attached to the chemical is gadolinium-DTPA or superparamagnetic iron oxide nanoparticles. 10. The method of claim 1,
wherein the method enables the accurate analysis of peripheral nerve lesions and the ability to distinguish axonotmetic lesions with greater self-regeneration potential from neurotmetic lesions that may require intervention. | 1,600 |
1,087 | 15,968,364 | 1,611 | An avermectin-based topical formulation is disclosed which is useful for prevention and treatment of head lice (Pediculus humanus capitis). This topical formulation may be formulated as a shampoo-condition which comprises an effective amount of avermectin, solubilizers, suspending agents, preservatives, nonionic surfactants, humectants, a silicone compound, and water. Also disclosed are methods of using the topical formulations disclosed within this specification to treat either a susceptible or treatment-resistant strain of lice, as well as uses in the manufacture of a medicament for treating or preventing a lice infestation from a susceptible or treatment-resistant strain in a human patient. | 1.-20. (canceled) 21. A method for treatment or prophylaxis of a lice infestation, the method comprising:
(a) applying a pediculicidal formulation to a human scalp infested with lice; wherein the pediculicidal formulation comprises:
about 0.1% to about 2.0% by weight ivermectin;
less than about 30% of a pharmaceutically acceptable glycol;
at least one suspending agent;
about 10 to about 35% of at least one non-iononic surfactant; and
water,
(b) maintaining the topical pediculicidal formulation on the scalp for about 1 to about 60 minutes to provide treated lice; and (c) removing the pediculicidal formulation from the scalp, wherein the treated lice have a faster mortality response than lice treated with unformulated 0.5% ivermectin. 22. The method of claim 21, wherein the pediculicidal formulation is packaged as a single dose. 23. The method of claim 21, wherein the pediculicidal formulation comprises about 0.5% by weight ivermectin. 24. The method of claim 21, wherein the at least one suspending agent is selected from the group consisting of fatty oils, fats and combinations thereof. 25. The method of claim 21, wherein the pediculicidal formulation comprises at least two suspending agents, wherein the at least two suspending agents comprise a fatty oil and a fat. 26. The method of claim 25, wherein the fatty oil is olive oil. 27. The method of claim 5, wherein the fat is shea butter. 28. The method of claim 21, wherein the pediculicidal formulation comprises at least two non-ionic surfactants. 29. The method of claim 28, wherein the at least two non-ionic surfactants are selected from the group consisting of fatty alcohols, sorbitan tristearate and combinations thereof. 30. The method of claim 29, wherein the fatty alcohol is selected from the group consisting of oleyl alcohol, lanolin alcohol and combinations thereof. 31. The method of claim 1, wherein the pediculicidal formulation comprises less than about 25% of a pharmaceutically acceptable glycol. 32. The method of claim 21, wherein the pediculicidal formulation comprises less than about 20% of a pharmaceutically acceptable glycol. 33. The method of claim 21, wherein the pediculicidal formulation comprises less than about 15% of a pharmaceutically acceptable glycol. 34. The method of claim 21, wherein the pediculicidal formulation comprises less than about 10% of a pharmaceutically acceptable glycol. 35. The method of claim 21, wherein the pediculicidal formulation has the texture of a gel or shampoo-conditioner. 36. The method of claim 21, wherein the pediculicidal formulation has the texture of a lotion. 37. The method of claim 21, wherein the non-ionic surfactant is selected from stearyl acetate, oleyl acetate and combinations thereof. 38. The method of claim 21, wherein the pediculicidal formulation is maintained on the scalp for about 3 to about 60 minutes. 39. The method of claim 21, wherein the pediculicidal formulation is maintained on the scalp for about 10 minutes. 40. The method of claim 21, further comprising (d) removing the pediculicidal formulation from the scalp by rinsing. | An avermectin-based topical formulation is disclosed which is useful for prevention and treatment of head lice (Pediculus humanus capitis). This topical formulation may be formulated as a shampoo-condition which comprises an effective amount of avermectin, solubilizers, suspending agents, preservatives, nonionic surfactants, humectants, a silicone compound, and water. Also disclosed are methods of using the topical formulations disclosed within this specification to treat either a susceptible or treatment-resistant strain of lice, as well as uses in the manufacture of a medicament for treating or preventing a lice infestation from a susceptible or treatment-resistant strain in a human patient.1.-20. (canceled) 21. A method for treatment or prophylaxis of a lice infestation, the method comprising:
(a) applying a pediculicidal formulation to a human scalp infested with lice; wherein the pediculicidal formulation comprises:
about 0.1% to about 2.0% by weight ivermectin;
less than about 30% of a pharmaceutically acceptable glycol;
at least one suspending agent;
about 10 to about 35% of at least one non-iononic surfactant; and
water,
(b) maintaining the topical pediculicidal formulation on the scalp for about 1 to about 60 minutes to provide treated lice; and (c) removing the pediculicidal formulation from the scalp, wherein the treated lice have a faster mortality response than lice treated with unformulated 0.5% ivermectin. 22. The method of claim 21, wherein the pediculicidal formulation is packaged as a single dose. 23. The method of claim 21, wherein the pediculicidal formulation comprises about 0.5% by weight ivermectin. 24. The method of claim 21, wherein the at least one suspending agent is selected from the group consisting of fatty oils, fats and combinations thereof. 25. The method of claim 21, wherein the pediculicidal formulation comprises at least two suspending agents, wherein the at least two suspending agents comprise a fatty oil and a fat. 26. The method of claim 25, wherein the fatty oil is olive oil. 27. The method of claim 5, wherein the fat is shea butter. 28. The method of claim 21, wherein the pediculicidal formulation comprises at least two non-ionic surfactants. 29. The method of claim 28, wherein the at least two non-ionic surfactants are selected from the group consisting of fatty alcohols, sorbitan tristearate and combinations thereof. 30. The method of claim 29, wherein the fatty alcohol is selected from the group consisting of oleyl alcohol, lanolin alcohol and combinations thereof. 31. The method of claim 1, wherein the pediculicidal formulation comprises less than about 25% of a pharmaceutically acceptable glycol. 32. The method of claim 21, wherein the pediculicidal formulation comprises less than about 20% of a pharmaceutically acceptable glycol. 33. The method of claim 21, wherein the pediculicidal formulation comprises less than about 15% of a pharmaceutically acceptable glycol. 34. The method of claim 21, wherein the pediculicidal formulation comprises less than about 10% of a pharmaceutically acceptable glycol. 35. The method of claim 21, wherein the pediculicidal formulation has the texture of a gel or shampoo-conditioner. 36. The method of claim 21, wherein the pediculicidal formulation has the texture of a lotion. 37. The method of claim 21, wherein the non-ionic surfactant is selected from stearyl acetate, oleyl acetate and combinations thereof. 38. The method of claim 21, wherein the pediculicidal formulation is maintained on the scalp for about 3 to about 60 minutes. 39. The method of claim 21, wherein the pediculicidal formulation is maintained on the scalp for about 10 minutes. 40. The method of claim 21, further comprising (d) removing the pediculicidal formulation from the scalp by rinsing. | 1,600 |
1,088 | 15,103,208 | 1,617 | Compositions comprising a substrate coated with a sulfonylurea herbicide, and methods for making and using such compositions, are generally described herein. | 1. A composition comprising a substrate, a sulfonylurea herbicide and a carrier. 2. The composition of claim 1, wherein the carrier comprises polybutene, white mineral oil, mineral seal oil, hexylene glycol, or combinations thereof. 3. The composition of claim 1 or 2, wherein the carrier comprises 0.01-10.00% by weight of the composition. 4. The composition of any of claims 1-3, wherein the substrate comprises a stabilizing compound. 5. The composition of claim 4, wherein the stabilizing compound comprises cellulose fiber. 6. The composition of claim 5, wherein the cellulose fiber comprises BIODAC®. 7. The composition of any of the above claims, wherein the substrate comprises a fertilizer. 8. The composition of claim 7, wherein the fertilizer is methylene urea, coated urea, uncoated urea, or combinations thereof. 9. The composition of any of claims 1-8, wherein the sulfonylurea herbicide comprises metsulfuron-methyl. 10. The composition of any of claims 1-9, wherein the composition has less than 10% degradation of metsulfuron-methyl over a period of 9 months. 11. The composition of any of claims 1-9, wherein the composition has less than 10% degradation when stored at a temperature of 50° C. or less for one month. 12. The composition of any of claims 1-9, wherein the composition has less than 10% degradation when stored at a temperature of 40° C. or less for two months. 13. The composition of any of claims 1-9, wherein the composition has less than 10% degradation when stored at a temperature of 35° C. or less for three months. 14. The composition of any of claims 1-9, wherein the composition has less than 10% degradation when stored at room temperature or less for 24 months. 15. The composition of any of claims 1-14, wherein the composition has a pH range of 7.0-10.0. 16. The composition of any of claims 1-15, further comprising a water-absorbent component. 17. The composition of claim 16, wherein the water-absorbent component is magnesium sulfate, clay, specialty silica, precipitate silica, calcium silicate, calcium sulfate, or combinations thereof. 18. The composition of claim 17, wherein the specialty silica comprises SIPERNAT D-17®. 19. The composition of claim 17, wherein the precipitate silica comprises HISIL®. 20. The composition of claims 16-19, wherein the water-absorbent component comprises 0.01-10.0% by weight of the compound. 21. The composition of any of claims 1-20, wherein the composition has a moisture level of 0-2%. 22. The composition of any of claims 1-21, wherein the substrate is coated with the sulfonylurea herbicide and the carrier. 23. The composition of claim 7, wherein the fertilizer is coated with the sulfonylurea herbicide and the carrier. 24. The composition of claim 23, wherein the substrate further comprises a stabilizing compound. 25. The composition of claim 24, wherein the stabilizing compound is uncoated by the sulfonylurea herbicide and the carrier. 26. A container or bag comprising the composition of any one of claims 1-25. 27. A method of making an herbicidal composition comprising suspending a sulfonylurea herbicide in a carrier to obtain a liquid suspension and spraying the liquid suspension onto a substrate, wherein the carrier is a solvent. 28. The method of claim 27, wherein the substrate comprises a stabilizing compound. 29. The method of claim 28, wherein the stabilizing compound comprises a cellulose fiber. 30. The method of claim 29, wherein the cellulose fiber comprises BIODAC®. 31. The method of any of claims 27-30, wherein the substrate further comprises a granular fertilizer. 32. The method of claim 31, wherein the granular fertilizer comprises methylene urea, coated urea, uncoated urea, or combinations thereof. 33. The method of any of claims 27-32, wherein the sulfonylurea herbicide is metsulfuron-methyl. 34. The method of any of claims 27-33, wherein the solvent comprises polybutene, white mineral oil, mineral seal oil, hexylene glycol, or combinations thereof. 35. The method of any of claims 27-34, wherein the carrier comprises 0.01-10.00% by weight of the composition. 36. The method of any of claims 27-35, further comprising adding a water-absorbent compound after spraying the liquid suspension onto the substrate or adding a water-absorbent compound before spraying the liquid suspension onto the substrate. 37. The method of claim 36, wherein the water-absorbent compound comprises magnesium sulfate, clay, specialty silica, precipitate silica, calcium silicate, calcium sulfate, or combinations thereof. 38. The method of claim 36 or 37, wherein the water-absorbent compound comprises 0.01-10.0% by weight of the compound. 39. The method of any of claims 27-38, wherein the substrate is dried to a moisture content of less than 2%. 40. The method of any of claims 27-39, wherein the herbicidal composition has a moisture level of 0-2%. 41. The method of any of claims 27-40, wherein the substrate has a pH of 7.0-10.0. 42. The method of any of claims 27-41, wherein the sulfonylurea herbicide suspended in a carrier is distributed substantially equally onto the substrate. 43. A composition comprising a substrate comprising a stabilizing compound, wherein the substrate is coated with a sulfonylurea herbicide and a hydrophobic solvent, prepared by a process comprising the steps of:
(a) suspending the sulfonylurea herbicide in a hydrophobic solvent; and (b) spraying the compound of (a) onto the substrate. 44. The composition prepared by the process of claim 43, the process further comprising the step of adding a water-absorbent compound after the compound of (a) has been sprayed onto the substrate. 45. The composition prepared by the process of claim 43, the process further comprising the step of adding a water-absorbent compound to the substrate before the compound of (a) has been sprayed onto the substrate. 46. A composition comprising a substrate comprising a fertilizer, wherein the substrate is coated with a sulfonylurea herbicide and a hydrophobic solvent, prepared by a process comprising the steps of:
(a) suspending the sulfonylurea herbicide in a hydrophobic solvent; and (b) spraying the compound of (a) onto the substrate. 47. The composition prepared by the process of claim 46, the process further comprising the step of adding a water-absorbent compound after the compound of (a) has been sprayed onto the substrate. 48. The composition prepared by the process of claim 46, the process further comprising the step of adding a water-absorbent compound to the substrate before the compound of (a) has been sprayed onto the substrate. 49. A composition comprising a substrate comprising a fertilizer and a stabilizing compound, wherein the substrate is coated with a sulfonylurea herbicide and a hydrophobic solvent, prepared by a process comprising the steps of:
(a) suspending the sulfonylurea herbicide in a hydrophobic solvent; (b) mixing and/or blending the fertilizer and stabilizing compound to create a substrate; and (c) spraying the compound of (a) onto the substrate of (b). 50. The composition prepared by the process of claim 49, the process further comprising the step of adding a water-absorbent compound after the compound of (a) has been sprayed onto the substrate. 51. The composition prepared by the process of claim 49, the process further comprising the step of adding a water-absorbent compound to the substrate before the compound of (a) has been sprayed onto the substrate. 52. The composition prepared by the process of any of claims 45-51, the process further comprising the step of drying the fertilizer prior to the creation of the substrate. 53. The composition prepared by the process of any of claims 45-51, the process further comprising the step of drying the fertilizer after the creation of the substrate. 54. A method for treating a lawn, comprising applying the composition of any of claims 1-25 to the lawn. 55. A method for treating a lawn, comprising applying the composition of any of claims 43-53 to the lawn. 56. The method of any claim 54 or 55, wherein the composition is contained in a bag and then applied to the lawn. 57. A lawn treated with the composition of any of claims 1-25. 58. The lawn of claim 57, comprising 3.13 lb of said composition per 1000 square feet of lawn. 59. A composition comprising: fertilizer, cellulose fibers, metsulfuron-methyl, and polybutene. 60. The composition of claim 59, wherein the cellulose fibers are BIODAC®. 61. The composition of claim 60, further comprising magnesium sulfate. 62. A composition comprising: (1) fertilizer in an amount of 85-95%, (2) BIODAC® in an amount of 8.8-9.8%, (3) metsulfuron-methyl in an amount of 0.0200-0.0230%, (4) polybutene in an amount of 0.700%, and (5) magnesium sulfate in an amount of 0.1900-0.2200%. 63. A composition comprising cellulose fibers, metsulfuron-methyl, and polybutene. 64. The composition of claim 63, wherein the cellulose fibers are BIODAC®. 65. The composition of claim 64, further comprising magnesium sulfate. 66. A composition comprising: (1) BIODAC® in an amount of 97.0-98.0%, (2) metsulfuron-methyl in an amount of 0.0300-0.0310%, and (3) polybutene in an amount of 2.400-2.500%. | Compositions comprising a substrate coated with a sulfonylurea herbicide, and methods for making and using such compositions, are generally described herein.1. A composition comprising a substrate, a sulfonylurea herbicide and a carrier. 2. The composition of claim 1, wherein the carrier comprises polybutene, white mineral oil, mineral seal oil, hexylene glycol, or combinations thereof. 3. The composition of claim 1 or 2, wherein the carrier comprises 0.01-10.00% by weight of the composition. 4. The composition of any of claims 1-3, wherein the substrate comprises a stabilizing compound. 5. The composition of claim 4, wherein the stabilizing compound comprises cellulose fiber. 6. The composition of claim 5, wherein the cellulose fiber comprises BIODAC®. 7. The composition of any of the above claims, wherein the substrate comprises a fertilizer. 8. The composition of claim 7, wherein the fertilizer is methylene urea, coated urea, uncoated urea, or combinations thereof. 9. The composition of any of claims 1-8, wherein the sulfonylurea herbicide comprises metsulfuron-methyl. 10. The composition of any of claims 1-9, wherein the composition has less than 10% degradation of metsulfuron-methyl over a period of 9 months. 11. The composition of any of claims 1-9, wherein the composition has less than 10% degradation when stored at a temperature of 50° C. or less for one month. 12. The composition of any of claims 1-9, wherein the composition has less than 10% degradation when stored at a temperature of 40° C. or less for two months. 13. The composition of any of claims 1-9, wherein the composition has less than 10% degradation when stored at a temperature of 35° C. or less for three months. 14. The composition of any of claims 1-9, wherein the composition has less than 10% degradation when stored at room temperature or less for 24 months. 15. The composition of any of claims 1-14, wherein the composition has a pH range of 7.0-10.0. 16. The composition of any of claims 1-15, further comprising a water-absorbent component. 17. The composition of claim 16, wherein the water-absorbent component is magnesium sulfate, clay, specialty silica, precipitate silica, calcium silicate, calcium sulfate, or combinations thereof. 18. The composition of claim 17, wherein the specialty silica comprises SIPERNAT D-17®. 19. The composition of claim 17, wherein the precipitate silica comprises HISIL®. 20. The composition of claims 16-19, wherein the water-absorbent component comprises 0.01-10.0% by weight of the compound. 21. The composition of any of claims 1-20, wherein the composition has a moisture level of 0-2%. 22. The composition of any of claims 1-21, wherein the substrate is coated with the sulfonylurea herbicide and the carrier. 23. The composition of claim 7, wherein the fertilizer is coated with the sulfonylurea herbicide and the carrier. 24. The composition of claim 23, wherein the substrate further comprises a stabilizing compound. 25. The composition of claim 24, wherein the stabilizing compound is uncoated by the sulfonylurea herbicide and the carrier. 26. A container or bag comprising the composition of any one of claims 1-25. 27. A method of making an herbicidal composition comprising suspending a sulfonylurea herbicide in a carrier to obtain a liquid suspension and spraying the liquid suspension onto a substrate, wherein the carrier is a solvent. 28. The method of claim 27, wherein the substrate comprises a stabilizing compound. 29. The method of claim 28, wherein the stabilizing compound comprises a cellulose fiber. 30. The method of claim 29, wherein the cellulose fiber comprises BIODAC®. 31. The method of any of claims 27-30, wherein the substrate further comprises a granular fertilizer. 32. The method of claim 31, wherein the granular fertilizer comprises methylene urea, coated urea, uncoated urea, or combinations thereof. 33. The method of any of claims 27-32, wherein the sulfonylurea herbicide is metsulfuron-methyl. 34. The method of any of claims 27-33, wherein the solvent comprises polybutene, white mineral oil, mineral seal oil, hexylene glycol, or combinations thereof. 35. The method of any of claims 27-34, wherein the carrier comprises 0.01-10.00% by weight of the composition. 36. The method of any of claims 27-35, further comprising adding a water-absorbent compound after spraying the liquid suspension onto the substrate or adding a water-absorbent compound before spraying the liquid suspension onto the substrate. 37. The method of claim 36, wherein the water-absorbent compound comprises magnesium sulfate, clay, specialty silica, precipitate silica, calcium silicate, calcium sulfate, or combinations thereof. 38. The method of claim 36 or 37, wherein the water-absorbent compound comprises 0.01-10.0% by weight of the compound. 39. The method of any of claims 27-38, wherein the substrate is dried to a moisture content of less than 2%. 40. The method of any of claims 27-39, wherein the herbicidal composition has a moisture level of 0-2%. 41. The method of any of claims 27-40, wherein the substrate has a pH of 7.0-10.0. 42. The method of any of claims 27-41, wherein the sulfonylurea herbicide suspended in a carrier is distributed substantially equally onto the substrate. 43. A composition comprising a substrate comprising a stabilizing compound, wherein the substrate is coated with a sulfonylurea herbicide and a hydrophobic solvent, prepared by a process comprising the steps of:
(a) suspending the sulfonylurea herbicide in a hydrophobic solvent; and (b) spraying the compound of (a) onto the substrate. 44. The composition prepared by the process of claim 43, the process further comprising the step of adding a water-absorbent compound after the compound of (a) has been sprayed onto the substrate. 45. The composition prepared by the process of claim 43, the process further comprising the step of adding a water-absorbent compound to the substrate before the compound of (a) has been sprayed onto the substrate. 46. A composition comprising a substrate comprising a fertilizer, wherein the substrate is coated with a sulfonylurea herbicide and a hydrophobic solvent, prepared by a process comprising the steps of:
(a) suspending the sulfonylurea herbicide in a hydrophobic solvent; and (b) spraying the compound of (a) onto the substrate. 47. The composition prepared by the process of claim 46, the process further comprising the step of adding a water-absorbent compound after the compound of (a) has been sprayed onto the substrate. 48. The composition prepared by the process of claim 46, the process further comprising the step of adding a water-absorbent compound to the substrate before the compound of (a) has been sprayed onto the substrate. 49. A composition comprising a substrate comprising a fertilizer and a stabilizing compound, wherein the substrate is coated with a sulfonylurea herbicide and a hydrophobic solvent, prepared by a process comprising the steps of:
(a) suspending the sulfonylurea herbicide in a hydrophobic solvent; (b) mixing and/or blending the fertilizer and stabilizing compound to create a substrate; and (c) spraying the compound of (a) onto the substrate of (b). 50. The composition prepared by the process of claim 49, the process further comprising the step of adding a water-absorbent compound after the compound of (a) has been sprayed onto the substrate. 51. The composition prepared by the process of claim 49, the process further comprising the step of adding a water-absorbent compound to the substrate before the compound of (a) has been sprayed onto the substrate. 52. The composition prepared by the process of any of claims 45-51, the process further comprising the step of drying the fertilizer prior to the creation of the substrate. 53. The composition prepared by the process of any of claims 45-51, the process further comprising the step of drying the fertilizer after the creation of the substrate. 54. A method for treating a lawn, comprising applying the composition of any of claims 1-25 to the lawn. 55. A method for treating a lawn, comprising applying the composition of any of claims 43-53 to the lawn. 56. The method of any claim 54 or 55, wherein the composition is contained in a bag and then applied to the lawn. 57. A lawn treated with the composition of any of claims 1-25. 58. The lawn of claim 57, comprising 3.13 lb of said composition per 1000 square feet of lawn. 59. A composition comprising: fertilizer, cellulose fibers, metsulfuron-methyl, and polybutene. 60. The composition of claim 59, wherein the cellulose fibers are BIODAC®. 61. The composition of claim 60, further comprising magnesium sulfate. 62. A composition comprising: (1) fertilizer in an amount of 85-95%, (2) BIODAC® in an amount of 8.8-9.8%, (3) metsulfuron-methyl in an amount of 0.0200-0.0230%, (4) polybutene in an amount of 0.700%, and (5) magnesium sulfate in an amount of 0.1900-0.2200%. 63. A composition comprising cellulose fibers, metsulfuron-methyl, and polybutene. 64. The composition of claim 63, wherein the cellulose fibers are BIODAC®. 65. The composition of claim 64, further comprising magnesium sulfate. 66. A composition comprising: (1) BIODAC® in an amount of 97.0-98.0%, (2) metsulfuron-methyl in an amount of 0.0300-0.0310%, and (3) polybutene in an amount of 2.400-2.500%. | 1,600 |
1,089 | 15,538,691 | 1,641 | The present invention generally relates to method for forming an immunolabeling complex, the immunolabeling complex comprising a labeled monovalent biotin-binding composition. The invention also related to the use of the antibody-reporter molecule complex for detecting a target in a sample. | 1. A method for forming an immunolabeling complex, wherein the method comprises:
selecting a first biotinylated primary antibody; selecting a first monovalent biotin-binding composition labeled with a first reporter element, and mixing, in a reaction vessel, the first biotinylated primary antibody with the first labeled monovalent biotin-binding composition. 2. The method according to claim 1, wherein mixing the first biotinylated primary antibody with the first labeled monovalent biotin-binding composition attaches the first reporter element to the first biotinylated primary antibody. 3. The method according to claim 1, wherein the immunolabeling complex is to be subsequently used in contacting with a sample. 4. The method according to claim 1, further comprising:
contacting the immunolabeling complex with the sample. 5. The method according to claim 1, wherein the sample is different from the first biotinylated primary antibody. 6. The method according to claim 1, wherein the immunolabeling complex is to be subsequently used in contacting with a sample, the sample being different from the first biotinylated primary antibody. 7. The method according to claim 1, further comprising:
providing an instruction to use the immunolabeling complex for immunolabeling of a sample. 8. The method according to claim 7, wherein selecting the first biotinylated primary antibody and selecting the first monovalent biotin-binding composition is performed separated from mixing the first biotinylated primary antibody with the first labeled monovalent biotin-binding composition. 9. The method according to claim 1, wherein first labeled monovalent biotin binding composition is based on at least one of streptavidin, avidin, traptavidin, captavidin, tamavidin, bradavidin, neutravidin, and rhizavidin. 10. The method according to claim 1, wherein the method further comprises adding a blocking agent to block unbound portions of the monovalent biotin-binding composition after the first immunolabeling complex has been formed. 11. The method according to claim 1, wherein the reporter element is selected from a group comprising a fluorochrome, an enzyme, a peptide, quantum dots, and a transition metal. 12. The method according to claim 11, wherein the first monovalent biotin-binding composition is conjugated with the reporter element. 13. The method according to claim 1, wherein the ratio between the first monovalent biotin-binding composition and the first biotinylated primary antibody is selected such that the risk of the first monovalent biotin-binding composition blocking active biding sites of the first biotinylated primary antibody is reduced. 14. The method according to claim 1, wherein the reporter element is an oligonucleotide. 15. The method according to claim 1, wherein the reporter element is an antigen. 16. The method according to claim 1, wherein the monovalent biotin-binding composition is modified to increase the number of conjugation sites, allowing attachment of an increased number of reporter elements. 17. The method according to claim 1, wherein the monovalent biotin-binding composition is conjugated with macromolecules containing a plurality of molecules of a reporter element. 18. A method for detecting a target molecule in a sample, wherein the method comprises:
contacting a first immunolabeling complex formed according to any one of the preceding claims with the sample; incubating the sample for a time sufficient to permit the first immunolabeling complex to selectively bind to the target; and detecting the target. 19. The method according to claim 18, wherein the first reporter element is a fluorochrome and detecting the target comprises illuminating the immunolabeling complex. 20. The method according to claim 18, further comprising:
contacting a second immunolabeling complex formed according to claim 1 with the sample, and incubating the sample for a time sufficient to permit the second immunolabeling complex to selectively bind to the target, wherein the second immunolabeling complex comprising a second biotinylated primary antibody mixed with a second monovalent biotin binding composition, the second biotinylated primary antibody being different from the first biotinylated primary antibody, and the second monovalent biotin binding composition being labeled with a second reporter molecule being different from the first reporter molecule. 21. A kit of immunolabeling complexes, comprising:
a first immunolabeling complex formed according to claim 1; and a second immunolabeling complex formed according to claim 1, wherein the second immunolabeling complex comprising a second biotinylated primary antibody mixed with a second monovalent biotin-binding composition, the second biotinylated primary antibody being different from the first biotinylated primary antibody, and the second monovalent biotin-binding composition being labeled with a second reporter molecule being different from the first reporter molecule. 22. A kit of an immunolabeling complex and an enhancer step, comprising:
an immunolabeling complex formed according to claim 1; and at least one first enhancer component. wherein the monovalent biotin-binding composition comprises a first set of at least one of antigens and oligonucleotides that are recognized by the first enhancer component, and the first enhancer component is conjugated with reporter elements, or the first enhancer component is conjugated with a second set of at least one of antigens and oligonucleotide that are recognized by a second enhancer component that is conjugated with reporter elements, wherein the second set is different from the first set. | The present invention generally relates to method for forming an immunolabeling complex, the immunolabeling complex comprising a labeled monovalent biotin-binding composition. The invention also related to the use of the antibody-reporter molecule complex for detecting a target in a sample.1. A method for forming an immunolabeling complex, wherein the method comprises:
selecting a first biotinylated primary antibody; selecting a first monovalent biotin-binding composition labeled with a first reporter element, and mixing, in a reaction vessel, the first biotinylated primary antibody with the first labeled monovalent biotin-binding composition. 2. The method according to claim 1, wherein mixing the first biotinylated primary antibody with the first labeled monovalent biotin-binding composition attaches the first reporter element to the first biotinylated primary antibody. 3. The method according to claim 1, wherein the immunolabeling complex is to be subsequently used in contacting with a sample. 4. The method according to claim 1, further comprising:
contacting the immunolabeling complex with the sample. 5. The method according to claim 1, wherein the sample is different from the first biotinylated primary antibody. 6. The method according to claim 1, wherein the immunolabeling complex is to be subsequently used in contacting with a sample, the sample being different from the first biotinylated primary antibody. 7. The method according to claim 1, further comprising:
providing an instruction to use the immunolabeling complex for immunolabeling of a sample. 8. The method according to claim 7, wherein selecting the first biotinylated primary antibody and selecting the first monovalent biotin-binding composition is performed separated from mixing the first biotinylated primary antibody with the first labeled monovalent biotin-binding composition. 9. The method according to claim 1, wherein first labeled monovalent biotin binding composition is based on at least one of streptavidin, avidin, traptavidin, captavidin, tamavidin, bradavidin, neutravidin, and rhizavidin. 10. The method according to claim 1, wherein the method further comprises adding a blocking agent to block unbound portions of the monovalent biotin-binding composition after the first immunolabeling complex has been formed. 11. The method according to claim 1, wherein the reporter element is selected from a group comprising a fluorochrome, an enzyme, a peptide, quantum dots, and a transition metal. 12. The method according to claim 11, wherein the first monovalent biotin-binding composition is conjugated with the reporter element. 13. The method according to claim 1, wherein the ratio between the first monovalent biotin-binding composition and the first biotinylated primary antibody is selected such that the risk of the first monovalent biotin-binding composition blocking active biding sites of the first biotinylated primary antibody is reduced. 14. The method according to claim 1, wherein the reporter element is an oligonucleotide. 15. The method according to claim 1, wherein the reporter element is an antigen. 16. The method according to claim 1, wherein the monovalent biotin-binding composition is modified to increase the number of conjugation sites, allowing attachment of an increased number of reporter elements. 17. The method according to claim 1, wherein the monovalent biotin-binding composition is conjugated with macromolecules containing a plurality of molecules of a reporter element. 18. A method for detecting a target molecule in a sample, wherein the method comprises:
contacting a first immunolabeling complex formed according to any one of the preceding claims with the sample; incubating the sample for a time sufficient to permit the first immunolabeling complex to selectively bind to the target; and detecting the target. 19. The method according to claim 18, wherein the first reporter element is a fluorochrome and detecting the target comprises illuminating the immunolabeling complex. 20. The method according to claim 18, further comprising:
contacting a second immunolabeling complex formed according to claim 1 with the sample, and incubating the sample for a time sufficient to permit the second immunolabeling complex to selectively bind to the target, wherein the second immunolabeling complex comprising a second biotinylated primary antibody mixed with a second monovalent biotin binding composition, the second biotinylated primary antibody being different from the first biotinylated primary antibody, and the second monovalent biotin binding composition being labeled with a second reporter molecule being different from the first reporter molecule. 21. A kit of immunolabeling complexes, comprising:
a first immunolabeling complex formed according to claim 1; and a second immunolabeling complex formed according to claim 1, wherein the second immunolabeling complex comprising a second biotinylated primary antibody mixed with a second monovalent biotin-binding composition, the second biotinylated primary antibody being different from the first biotinylated primary antibody, and the second monovalent biotin-binding composition being labeled with a second reporter molecule being different from the first reporter molecule. 22. A kit of an immunolabeling complex and an enhancer step, comprising:
an immunolabeling complex formed according to claim 1; and at least one first enhancer component. wherein the monovalent biotin-binding composition comprises a first set of at least one of antigens and oligonucleotides that are recognized by the first enhancer component, and the first enhancer component is conjugated with reporter elements, or the first enhancer component is conjugated with a second set of at least one of antigens and oligonucleotide that are recognized by a second enhancer component that is conjugated with reporter elements, wherein the second set is different from the first set. | 1,600 |
1,090 | 16,100,158 | 1,633 | Methods for introducing exogenous material into a cell are provided, which include exposing the cell to a transient decrease in pressure in the presence of the exogenous material. Also provided are devices for performing the method of the invention. | 1. A method introducing an exogenous material into the cell, comprising:
inducing a first condition around the cell where there is an ambient pressure insufficient to lyse the cell; inducing a second condition around the cell where the second pressure is less than the ambient pressure; and inducing a third condition around the cell where the third pressure is greater than the second pressure, the third condition being induced while the cell is in an unsteady flow. 2. The method of claim 1, wherein the second condition is induced in less time than the third condition. 3. The method of claim 1, wherein the second condition is induced under unsteady flow conditions. 4. The method of claim 1, wherein the third condition is induced by pulsing the cell with an unsteady flow. 5. The method of claim 1, further comprising repeatedly pulsing the cell with an unsteady flow to facilitate delivery of the exogenous material into the cell. 6. The method of claim 1, wherein the cell is viable after the introduction of the exogenous material into the cell. 7. The method of claim 1, wherein the exogenous material is selected from the group consisting of an organic molecule, a physiologically acceptable organic molecule derivative, a biomolecule, a physiologically acceptable biomolecule derivative, a physiologically acceptable biomolecule analogue, an inorganic molecule, physiologically acceptable inorganic molecule derivative, a quantum dot, a carbon nanotube, a nanoparticle, and a gold particle. 8. The method of claim 7, wherein the biomolecule is selected from the group consisting of a carbohydrate, a lipid, an amino acid, a peptide, a protein, a nucleotide, and a nucleic acid. 9. The method of claim 8, wherein the nucleic acid is selected from the group consisting of a deoxyribonucleic acid and a ribonucleic acid. 10. The method of claim 8, wherein the nucleic acid is included in an expression vector. 11. The method of claim 10, wherein the expression vector is a plasmid. 12. The method of claim 10, wherein the expression vector comprises at least one regulatory sequence. 13. The method of claim 12, wherein the at least one regulatory sequence is selected from the group consisting of an enhancer region and a promoter region. 14. A method for introducing exogenous material into a cell, comprising:
exposing the cell to a first pressure change under unsteady flow conditions to temporarily permeabilise a cell membrane without the cell becoming lysed; and exposing the cell to a second pressure change under unsteady flow conditions to introduce the exogenous material into the cell, the first pressure change being a negative pressure change, the second pressure change being a positive pressure change. 15. The method of claim 14, wherein the first pressure change decreases extracellular pressure relative to intracellular pressure of the cell. 16. The method of claim 14, wherein the second pressure change increases extracellular pressure relative to intracellular pressure of the cell. 17. The method of claim 14, wherein the second pressure change is induced by pulsing the cell with an unsteady flow. 18. The method of claim 14, further comprising repeatedly pulsing the cell with an unsteady flow to facilitate delivery of the exogenous material into the cell. 19. The method of claim 14, wherein the cell is viable after the introduction of the exogenous material into the cell. 20. The method of claim 14, wherein the exogenous material is selected from the group consisting of an organic molecule, a physiologically acceptable organic molecule derivative, a biomolecule, a physiologically acceptable biomolecule derivative, a physiologically acceptable biomolecule analogue, an inorganic molecule, physiologically acceptable inorganic molecule derivative, a quantum dot, a carbon nanotube, a nanoparticle, and a gold particle. 21. The method of claim 20, wherein the biomolecule is selected from the group consisting of a carbohydrate, a lipid, an amino acid, a peptide, a protein, a nucleotide, and a nucleic acid. 22. The method of claim 21, wherein the nucleic acid is selected from the group consisting of a deoxyribonucleic acid and a ribonucleic acid. 23. The method of claim 21, wherein the nucleic acid is included in an expression vector. 24. The method of claim 23, wherein the expression vector is a plasmid. 25. The method of claim 23, wherein the expression vector comprises at least one regulatory sequence. 26. The method of claim 25, wherein the at least one regulatory sequence is selected from the group consisting of an enhancer region and a promoter region. 27. A modified cell containing exogenous material introduced into the cell by the process of:
inducing a first condition around the cell where there is an ambient pressure insufficient to lyse the cell; inducing a second condition around the cell where the second pressure is less than the ambient pressure; and inducing a third condition around the cell where the third pressure is greater than the second pressure, the third condition being induced while the cell is in an unsteady flow. 28. A pharmaceutical composition comprising:
a cell modified by temporarily permeabilising a cell membrane and introducing an exogenous material into the cell while the cell is pulsed by an unsteady flow; and a pharmaceutically acceptable carrier. | Methods for introducing exogenous material into a cell are provided, which include exposing the cell to a transient decrease in pressure in the presence of the exogenous material. Also provided are devices for performing the method of the invention.1. A method introducing an exogenous material into the cell, comprising:
inducing a first condition around the cell where there is an ambient pressure insufficient to lyse the cell; inducing a second condition around the cell where the second pressure is less than the ambient pressure; and inducing a third condition around the cell where the third pressure is greater than the second pressure, the third condition being induced while the cell is in an unsteady flow. 2. The method of claim 1, wherein the second condition is induced in less time than the third condition. 3. The method of claim 1, wherein the second condition is induced under unsteady flow conditions. 4. The method of claim 1, wherein the third condition is induced by pulsing the cell with an unsteady flow. 5. The method of claim 1, further comprising repeatedly pulsing the cell with an unsteady flow to facilitate delivery of the exogenous material into the cell. 6. The method of claim 1, wherein the cell is viable after the introduction of the exogenous material into the cell. 7. The method of claim 1, wherein the exogenous material is selected from the group consisting of an organic molecule, a physiologically acceptable organic molecule derivative, a biomolecule, a physiologically acceptable biomolecule derivative, a physiologically acceptable biomolecule analogue, an inorganic molecule, physiologically acceptable inorganic molecule derivative, a quantum dot, a carbon nanotube, a nanoparticle, and a gold particle. 8. The method of claim 7, wherein the biomolecule is selected from the group consisting of a carbohydrate, a lipid, an amino acid, a peptide, a protein, a nucleotide, and a nucleic acid. 9. The method of claim 8, wherein the nucleic acid is selected from the group consisting of a deoxyribonucleic acid and a ribonucleic acid. 10. The method of claim 8, wherein the nucleic acid is included in an expression vector. 11. The method of claim 10, wherein the expression vector is a plasmid. 12. The method of claim 10, wherein the expression vector comprises at least one regulatory sequence. 13. The method of claim 12, wherein the at least one regulatory sequence is selected from the group consisting of an enhancer region and a promoter region. 14. A method for introducing exogenous material into a cell, comprising:
exposing the cell to a first pressure change under unsteady flow conditions to temporarily permeabilise a cell membrane without the cell becoming lysed; and exposing the cell to a second pressure change under unsteady flow conditions to introduce the exogenous material into the cell, the first pressure change being a negative pressure change, the second pressure change being a positive pressure change. 15. The method of claim 14, wherein the first pressure change decreases extracellular pressure relative to intracellular pressure of the cell. 16. The method of claim 14, wherein the second pressure change increases extracellular pressure relative to intracellular pressure of the cell. 17. The method of claim 14, wherein the second pressure change is induced by pulsing the cell with an unsteady flow. 18. The method of claim 14, further comprising repeatedly pulsing the cell with an unsteady flow to facilitate delivery of the exogenous material into the cell. 19. The method of claim 14, wherein the cell is viable after the introduction of the exogenous material into the cell. 20. The method of claim 14, wherein the exogenous material is selected from the group consisting of an organic molecule, a physiologically acceptable organic molecule derivative, a biomolecule, a physiologically acceptable biomolecule derivative, a physiologically acceptable biomolecule analogue, an inorganic molecule, physiologically acceptable inorganic molecule derivative, a quantum dot, a carbon nanotube, a nanoparticle, and a gold particle. 21. The method of claim 20, wherein the biomolecule is selected from the group consisting of a carbohydrate, a lipid, an amino acid, a peptide, a protein, a nucleotide, and a nucleic acid. 22. The method of claim 21, wherein the nucleic acid is selected from the group consisting of a deoxyribonucleic acid and a ribonucleic acid. 23. The method of claim 21, wherein the nucleic acid is included in an expression vector. 24. The method of claim 23, wherein the expression vector is a plasmid. 25. The method of claim 23, wherein the expression vector comprises at least one regulatory sequence. 26. The method of claim 25, wherein the at least one regulatory sequence is selected from the group consisting of an enhancer region and a promoter region. 27. A modified cell containing exogenous material introduced into the cell by the process of:
inducing a first condition around the cell where there is an ambient pressure insufficient to lyse the cell; inducing a second condition around the cell where the second pressure is less than the ambient pressure; and inducing a third condition around the cell where the third pressure is greater than the second pressure, the third condition being induced while the cell is in an unsteady flow. 28. A pharmaceutical composition comprising:
a cell modified by temporarily permeabilising a cell membrane and introducing an exogenous material into the cell while the cell is pulsed by an unsteady flow; and a pharmaceutically acceptable carrier. | 1,600 |
1,091 | 15,245,426 | 1,641 | The present invention relates to a method for detecting a target substance that is useful for detection of a glycoprotein and the like comprising a specific sugar chain, a reagent for detecting a target substance, a carrier and a method for manufacturing the carrier. | 1. A reagent for detecting a target substance comprising a sugar chain that binds with lectin of Wisteria floribunda (WFA),
wherein the reagent comprises a dimeric WFA immobilized carrier comprising a solid phase carrier and a dimeric WFA, and the dimeric WFA is immobilized on the solid phase carrier. 2. The reagent according to claim 1, wherein the dimeric WFA is composed of two subunits. 3. The reagent according to claim 1, wherein the dimeric WFA is immobilized on the solid phase carrier via biotin and biotin-binding protein. 4. The reagent according to claim 3, wherein the dimeric WFA is biotinylated dimeric WFA, and the biotin-binding protein is immobilized on the solid phase carrier, and the biotinylated dimeric WFA is immobilized on the solid phase carrier via the biotin-binding protein. 5. The reagent according to claim 1, wherein the solid phase carrier is a magnetic particle. 6. A method for detecting a target substance comprising a sugar chain that binds with lectin of Wisteria floribunda (WFA), comprising the steps of:
(A) bringing a dimeric WFA immobilized carrier comprising a solid phase carrier and a dimeric WFA which is immobilized on the solid phase carrier, a sample comprising the target substance, and a labeling substance that specifically binds with the target substance into contact with each other to form a complex comprising the dimeric WFA, the target substance and the labeling substance on the solid phase carrier, and (B) measuring the labeling substance in the complex obtained in the step (A) to detect the target substance. 7. The method according to claim 6, wherein the labeling substance is a labeled antibody that specifically binds with the target substance. 8. The method according to claim 6, wherein the dimeric WFA is composed of two subunits. 9. The method according to claim 6, wherein the dimeric WFA is immobilized on the solid phase carrier via biotin and biotin-binding protein. 10. The method according to claim 9, wherein the dimeric WFA is biotinylated dimeric WFA, and the biotin-binding protein is immobilized on the solid phase carrier, and the biotinylated dimeric WFA is immobilized on the solid phase carrier via the biotin-binding protein. 11. The method according to claim 6, wherein the solid phase carrier is a magnetic particle. 12. A method for manufacturing a carrier to be used for detecting a target substance comprising a sugar chain that binds with lectin of Wisteria floribunda (WFA), comprising the steps of:
(I) dimerizing WFA to obtain dimeric WFA, and (II) immobilizing the dimeric WFA obtained in the step (I) on a solid phase carrier to obtain a dimeric WFA immobilized carrier. 13. The method according to claim 12, wherein the dimeric WFA is composed of two subunits. 14. The method according to claim 12, wherein biotinylated dimeric WFA is prepared by mixing a solution comprising a cross-linking agent comprising biotin, and WFA in the step (I). 15. The method according to claim 14, wherein a carrier on which biotin-binding protein is immobilized is used as the solid phase carrier in the step (II), and the dimeric WFA is immobilized on the solid phase carrier by letting the biotin-binding protein in the carrier and the biotinylated dimeric WFA bind with each other. 16. The method according to claim 14, wherein the dimeric WFA is obtained by bringing the solution comprising the cross-linking agent into contact with WFA so that the molar ratio of WFA/cross-linking agent is less than or equal to 1/10 (excluding 0) in the step (I). 17. The method according to claim 14, wherein the cross-linking agent is a cross-linking agent that forms a bridge together with an amino group in the dimeric WFA. 18. The method according to claim 14, wherein the cross-linking agent has at least one functional group selected from the group consisting of a N-hydroxysuccinimide ester group, an isothiocyano group, a chlorosulfone group, a chlorocarbonyl group, an oxyethylene group, a chloroalkyl group having 1 to 4 carbon atoms, an aldehyde group and a carboxyl group as a reactive group with the amino group in the dimeric WFA. 19. The method according to claim 18, wherein the biotin and the reactive group bind to each other via a spacer. 20. The method according to claim 12, wherein the solid phase carrier is a magnetic particle. | The present invention relates to a method for detecting a target substance that is useful for detection of a glycoprotein and the like comprising a specific sugar chain, a reagent for detecting a target substance, a carrier and a method for manufacturing the carrier.1. A reagent for detecting a target substance comprising a sugar chain that binds with lectin of Wisteria floribunda (WFA),
wherein the reagent comprises a dimeric WFA immobilized carrier comprising a solid phase carrier and a dimeric WFA, and the dimeric WFA is immobilized on the solid phase carrier. 2. The reagent according to claim 1, wherein the dimeric WFA is composed of two subunits. 3. The reagent according to claim 1, wherein the dimeric WFA is immobilized on the solid phase carrier via biotin and biotin-binding protein. 4. The reagent according to claim 3, wherein the dimeric WFA is biotinylated dimeric WFA, and the biotin-binding protein is immobilized on the solid phase carrier, and the biotinylated dimeric WFA is immobilized on the solid phase carrier via the biotin-binding protein. 5. The reagent according to claim 1, wherein the solid phase carrier is a magnetic particle. 6. A method for detecting a target substance comprising a sugar chain that binds with lectin of Wisteria floribunda (WFA), comprising the steps of:
(A) bringing a dimeric WFA immobilized carrier comprising a solid phase carrier and a dimeric WFA which is immobilized on the solid phase carrier, a sample comprising the target substance, and a labeling substance that specifically binds with the target substance into contact with each other to form a complex comprising the dimeric WFA, the target substance and the labeling substance on the solid phase carrier, and (B) measuring the labeling substance in the complex obtained in the step (A) to detect the target substance. 7. The method according to claim 6, wherein the labeling substance is a labeled antibody that specifically binds with the target substance. 8. The method according to claim 6, wherein the dimeric WFA is composed of two subunits. 9. The method according to claim 6, wherein the dimeric WFA is immobilized on the solid phase carrier via biotin and biotin-binding protein. 10. The method according to claim 9, wherein the dimeric WFA is biotinylated dimeric WFA, and the biotin-binding protein is immobilized on the solid phase carrier, and the biotinylated dimeric WFA is immobilized on the solid phase carrier via the biotin-binding protein. 11. The method according to claim 6, wherein the solid phase carrier is a magnetic particle. 12. A method for manufacturing a carrier to be used for detecting a target substance comprising a sugar chain that binds with lectin of Wisteria floribunda (WFA), comprising the steps of:
(I) dimerizing WFA to obtain dimeric WFA, and (II) immobilizing the dimeric WFA obtained in the step (I) on a solid phase carrier to obtain a dimeric WFA immobilized carrier. 13. The method according to claim 12, wherein the dimeric WFA is composed of two subunits. 14. The method according to claim 12, wherein biotinylated dimeric WFA is prepared by mixing a solution comprising a cross-linking agent comprising biotin, and WFA in the step (I). 15. The method according to claim 14, wherein a carrier on which biotin-binding protein is immobilized is used as the solid phase carrier in the step (II), and the dimeric WFA is immobilized on the solid phase carrier by letting the biotin-binding protein in the carrier and the biotinylated dimeric WFA bind with each other. 16. The method according to claim 14, wherein the dimeric WFA is obtained by bringing the solution comprising the cross-linking agent into contact with WFA so that the molar ratio of WFA/cross-linking agent is less than or equal to 1/10 (excluding 0) in the step (I). 17. The method according to claim 14, wherein the cross-linking agent is a cross-linking agent that forms a bridge together with an amino group in the dimeric WFA. 18. The method according to claim 14, wherein the cross-linking agent has at least one functional group selected from the group consisting of a N-hydroxysuccinimide ester group, an isothiocyano group, a chlorosulfone group, a chlorocarbonyl group, an oxyethylene group, a chloroalkyl group having 1 to 4 carbon atoms, an aldehyde group and a carboxyl group as a reactive group with the amino group in the dimeric WFA. 19. The method according to claim 18, wherein the biotin and the reactive group bind to each other via a spacer. 20. The method according to claim 12, wherein the solid phase carrier is a magnetic particle. | 1,600 |
1,092 | 15,442,695 | 1,636 | The invention provides methods of genetically engineering a C1-fixing bacterium using a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas) (CRISPR/Cas) system. Preferably, the Cas protein is under the control of an inducible promoter. | 1. A method of genetically engineering a C1-fixing bacterium comprising introducing into a C1-fixing bacterium containing a DNA molecule comprising a target sequence an engineered, non-naturally occurring Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas) (CRISPR/Cas) system comprising one or more vectors comprising:
(a) a nucleotide sequence encoding a guide RNA that hybridizes with the target sequence and (b) a nucleotide sequence encoding a type-II Cas9 protein under the control of an inducible promoter. 2. The method of claim 1, wherein the CRISPR/Cas system further comprises on the one or more vectors:
(c) a nucleotide sequence comprising a 5′ homology arm that hybridizes upstream of the target sequence and a 3′ homology arm that hybridizes downstream of the target sequence, whereby the 5′ homology arm and the 3′ homology arm hybridize with the DNA molecule and homologous recombination occurs, resulting in the replacement of the target sequence with DNA located between the 5′ homology arm and the 3′ homology arm. 3. The method of claim 1, wherein the Cas9 is catalytically active. 4. The method of claim 1, wherein the Cas9 is nickase Cas9. 5. The method of claim 1, wherein the Cas9 is catalytically inactive. 6. The method of claim 1, wherein the Cas9 cleaves the DNA molecule in a region encoding a gene, whereby expression of the gene is decreased. 7. The method of claim 1, wherein the Cas9 blocks the DNA molecule in a region encoding a gene, whereby expression of the gene is decreased. 8. The method of claim 2, wherein the DNA located between the 5′ homology arm and the 3′ homology arm disrupts the DNA molecule in a region encoding a gene, whereby expression of the gene is decreased. 9. The method of claim 2, wherein the DNA located between the 5′ homology arm and the 3′ homology arm encodes an exogenous gene, whereby the homologous recombination inserts the exogenous gene into the DNA molecule. 10. The method of claim 9, wherein the C1-fixing bacterium expresses the exogenous gene. 11. The method of claim 1, wherein (a) and (b) are located on the same or different vectors. 12. The method of claim 2, wherein (a), (b), and (c) are located on the same or different vectors. 13. The method of claim 1, wherein the CRISPR/Cas system is derived from Streptococcus pyogenes or Streptococcus thermophilus. 14. The method of claim 1, wherein the inducible promotor is a tetracycline inducible promoter or a lactose inducible promoter. 15. The method of claim 14, wherein the tetracycline inducible promoter is tet3no or ip112. 16. The method of claim 1, wherein the C1-fixing bacterium is selected from the group consisting of Acetobacterium woodii, Alkalibaculum bacchii, Blautia producta, Butyribacterium methylotrophicum, Clostridium aceticum, Clostridium autoethanogenum, Clostridium carboxidivorans, Clostridium coskatii, Clostridium drakei, Clostridium formicoaceticum, Clostridium ljungdahlii, Clostridium magnum, Clostridium ragsdalei, Clostridium scatologenes, Eubacterium limosum, Moorella thermautotrophica, Moorella thermoacetica, Oxobacter pfennigii, Sporomusa ovata, Sporomusa silvacetica, Sporomusa sphaeroides, and Thermoanaerobacter kiuvi. 17. The method of claim 1, wherein the C1-fixing bacterium is Clostridium autoethanogenum, Clostridium ljungdahlii, or Clostridium ragsdalei. 18. The method of claim 1, wherein the C1-fixing bacterium is Clostridium autoethanogenum. | The invention provides methods of genetically engineering a C1-fixing bacterium using a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas) (CRISPR/Cas) system. Preferably, the Cas protein is under the control of an inducible promoter.1. A method of genetically engineering a C1-fixing bacterium comprising introducing into a C1-fixing bacterium containing a DNA molecule comprising a target sequence an engineered, non-naturally occurring Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas) (CRISPR/Cas) system comprising one or more vectors comprising:
(a) a nucleotide sequence encoding a guide RNA that hybridizes with the target sequence and (b) a nucleotide sequence encoding a type-II Cas9 protein under the control of an inducible promoter. 2. The method of claim 1, wherein the CRISPR/Cas system further comprises on the one or more vectors:
(c) a nucleotide sequence comprising a 5′ homology arm that hybridizes upstream of the target sequence and a 3′ homology arm that hybridizes downstream of the target sequence, whereby the 5′ homology arm and the 3′ homology arm hybridize with the DNA molecule and homologous recombination occurs, resulting in the replacement of the target sequence with DNA located between the 5′ homology arm and the 3′ homology arm. 3. The method of claim 1, wherein the Cas9 is catalytically active. 4. The method of claim 1, wherein the Cas9 is nickase Cas9. 5. The method of claim 1, wherein the Cas9 is catalytically inactive. 6. The method of claim 1, wherein the Cas9 cleaves the DNA molecule in a region encoding a gene, whereby expression of the gene is decreased. 7. The method of claim 1, wherein the Cas9 blocks the DNA molecule in a region encoding a gene, whereby expression of the gene is decreased. 8. The method of claim 2, wherein the DNA located between the 5′ homology arm and the 3′ homology arm disrupts the DNA molecule in a region encoding a gene, whereby expression of the gene is decreased. 9. The method of claim 2, wherein the DNA located between the 5′ homology arm and the 3′ homology arm encodes an exogenous gene, whereby the homologous recombination inserts the exogenous gene into the DNA molecule. 10. The method of claim 9, wherein the C1-fixing bacterium expresses the exogenous gene. 11. The method of claim 1, wherein (a) and (b) are located on the same or different vectors. 12. The method of claim 2, wherein (a), (b), and (c) are located on the same or different vectors. 13. The method of claim 1, wherein the CRISPR/Cas system is derived from Streptococcus pyogenes or Streptococcus thermophilus. 14. The method of claim 1, wherein the inducible promotor is a tetracycline inducible promoter or a lactose inducible promoter. 15. The method of claim 14, wherein the tetracycline inducible promoter is tet3no or ip112. 16. The method of claim 1, wherein the C1-fixing bacterium is selected from the group consisting of Acetobacterium woodii, Alkalibaculum bacchii, Blautia producta, Butyribacterium methylotrophicum, Clostridium aceticum, Clostridium autoethanogenum, Clostridium carboxidivorans, Clostridium coskatii, Clostridium drakei, Clostridium formicoaceticum, Clostridium ljungdahlii, Clostridium magnum, Clostridium ragsdalei, Clostridium scatologenes, Eubacterium limosum, Moorella thermautotrophica, Moorella thermoacetica, Oxobacter pfennigii, Sporomusa ovata, Sporomusa silvacetica, Sporomusa sphaeroides, and Thermoanaerobacter kiuvi. 17. The method of claim 1, wherein the C1-fixing bacterium is Clostridium autoethanogenum, Clostridium ljungdahlii, or Clostridium ragsdalei. 18. The method of claim 1, wherein the C1-fixing bacterium is Clostridium autoethanogenum. | 1,600 |
1,093 | 14,768,970 | 1,631 | A system including a device with at least one processor and memory storing computer-executable instructions that, when executed by the at least one processor, perform a method of identifying glycopeptides in a sample, the method including, analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides, identifying the glycopeptides in the sample based on the at least one identified portion; and analyzing at least one glycopeptide of the identified glycopeptides. | 1. At least one computer-readable storage medium storing computer-executable instructions that, when executed by at least one processor, perform a method of identifying glycopeptides in a sample, the method comprising:
analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides; and identifying the glycopeptides in the sample based on the at least one identified portion. 2. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
determining the at least one characteristic of mass spectra indicative of presence of glycopeptides. 3. The at least one computer-readable storage medium of claim 2, wherein:
determining the at least one characteristic comprises determining at least one glycopeptide-rich acquisition enhancement zone. 4. The at least one computer-readable storage medium of claim 2, wherein:
determining the at least one characteristic comprises analyzing a training data set comprising a plurality of mass spectra of peptides. 5. The at least one computer-readable storage medium of claim 1, wherein:
the at least one characteristic comprises at least one first range of a nominal mass and at least one second range of mass defect. 6. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
displaying on a user interface results of the identification of the glycopeptides in the sample. 7. The at least one computer-readable storage medium of claim 6, wherein:
displaying the results of the identification of the glycopeptides comprises displaying the results so that the glycopeptides in the sample are differentiated from peptides in the sample. 8. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
providing a representation of the results of the identification of the glycopeptides so that the representation is enabled to receive input indicating selection of at least one glycopeptide of the identified glycopeptides for further analysis. 9. The at least one computer-readable storage medium of claim 8, wherein the method further comprises:
further analyzing the at least one glycopeptide selected for the further analysis. 10. The at least one computer-readable storage medium of claim 1, wherein:
identifying the glycopeptides in the sample comprises identifying N-glycosylated glycopeptides. 11. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
providing results of the identification of the glycopeptides in the sample to a system configured to further analyze the identified glycopeptides. 12. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
further analyzing at least one of the identified glycopeptides. 13. The at least one computer-readable storage medium of claim 1, wherein the sample comprises a biological sample. 14. The at least one computer-readable storage medium of claim 13, wherein the biological sample is obtained from tissue, urine, blood, plasma, serum or saliva. 15. The at least one computer-readable storage medium of claim 2, wherein:
the at least one characteristic is determined for a protease used to generate a mixture of peptides and glycopeptides from the sample. 16. The at least one computer-readable storage medium of claim 1, wherein:
analyzing the mass spectrum comprises analyzing precursor ion data. 17. At least one computer-readable storage medium storing computer-executable instructions that, when executed by at least one processor, perform a method of identifying glycopeptides in a sample, the method comprising:
determining at least one characteristic of mass spectra indicative of presence of glycopeptides; analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having the at least one characteristic; and identifying the glycopeptides in the sample based on the at least one identified portion. 18. A computer-implemented method of identifying glycopeptides in a sample, the method comprising:
with at least one processor:
analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides; and
identifying the glycopeptides in the sample based on the at least one identified portion. 19. The method of claim 18, further comprising:
determining the at least one characteristic of mass spectra indicative of presence of glycopeptides. 20. The method of claim 19, wherein:
determining the at least one characteristic comprises determining at least one glycopeptide-rich acquisition enhancement zone. 21. The method of claim 19, wherein:
determining the at least one characteristic comprises analyzing a data set comprising a plurality of mass spectra of peptides to determine at least one first range of a nominal mass and at least one second range of mass defect indicative of presence of glycopeptides. 22. The method of claim 19, wherein:
determining the at least one characteristic comprises analyzing a training data set comprising a plurality of mass spectra of peptides. 23. The method of claim 18, wherein:
the at least one characteristic comprises at least one first range of a nominal mass and at least one second range of mass defect. 24. The method of claim 18, further comprising:
displaying on a user interface results of the identification of the glycopeptides in the sample. 25. The method of claim 24, wherein:
displaying the results of the identification of the glycopeptides comprises displaying the results so that the glycopeptides in the sample are differentiated from peptides in the sample. 26. The method of claim 18, further comprising:
providing a representation of the results of the identification of the glycopeptides so that the representation is enabled to receive input indicating selection of at least one glycopeptide of the identified glycopeptides for further analysis. 27. The method of claim 26, further comprising:
further analyzing the at least one glycopeptide selected for the further analysis. 28. The method of claim 27, wherein:
further analyzing the at least one glycopeptide selected for the further analysis comprises determining a site of glycosylation on the at least one glycopeptide. 29. The system of claim 28, wherein:
determining the site of glycosylation comprises determining a site of N-glycosylation on the at least one glycopeptide. 30. The method of claim 27, further comprising:
analyzing the at least one glycopeptide using tandem mass-spectrometry. 31. The method of claim 18, wherein:
identifying the glycopeptides in the sample comprises identifying N-glycosylated glycopeptides. 32. The method of claim 18, further comprising:
providing results of the identification of the glycopeptides in the sample to a system configured to further analyze the identified glycopeptides. 33. The method of claim 18, further comprising:
further analyzing at least one of the identified glycopeptides. 34. The method of claim 18, wherein the sample comprises a biological sample. 35. The method of claim 34, wherein the biological sample is obtained from tissue, urine, blood, plasma, serum or saliva. 36. The method of claim 18, wherein:
analyzing the mass spectrum comprises analyzing precursor ion data. 37. A device comprising at least one processor and memory storing computer-executable instructions that, when executed by the at least one processor, perform a method of identifying glycopeptides in a sample, the method comprising:
analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides; and identifying the glycopeptides in the sample based on the at least one identified portion. 38. The device of claim 37, wherein the method further comprises:
determining the at least one characteristic of mass spectra indicative of presence of glycopeptides. 39. The device of claim 38, wherein:
determining the at least one characteristic comprises determining at least one glycopeptide-rich acquisition enhancement zone. 40. The device of claim 38, wherein:
determining the at least one characteristic comprises analyzing a data set comprising a plurality of mass spectra of peptides to determine at least one first range of a nominal mass and at least one second range of mass defect indicative of presence of glycopeptides. 41. The device of claim 38, wherein:
determining the at least one characteristic comprises analyzing a training data set comprising a plurality of mass spectra of peptides. 42. A system comprising:
a device comprising at least one processor and memory storing computer-executable instructions that, when executed by the at least one processor, perform a method of identifying glycopeptides in a sample, the method comprising:
analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides;
identifying the glycopeptides in the sample based on the at least one identified portion; and
analyzing at least one glycopeptide of the identified glycopeptides. 43. The system of claim 42, wherein the method further comprises:
determining the at least one characteristic of mass spectra indicative of presence of glycopeptides. 44. The system of claim 43, wherein:
determining the at least one characteristic comprises determining at least one glycopeptide-rich acquisition enhancement zone. 45. The system of claim 43, wherein:
determining the at least one characteristic comprises analyzing a data set comprising a plurality of mass spectra of peptides to determine at least one first range of a nominal mass and at least one second range of mass defect indicative of presence of glycopeptides. 46. The system of claim 43, wherein:
determining the at least one characteristic comprises analyzing a training data set comprising a plurality of mass spectra of peptides. 47. The system of claim 42, wherein:
analyzing the at least one glycopeptide comprises determining a site of glycosylation on the at least one glycopeptide. 48. The system of claim 47, wherein:
determining the site of glycosylation comprises determining a site of N-glycosylation on the at least one glycopeptide. | A system including a device with at least one processor and memory storing computer-executable instructions that, when executed by the at least one processor, perform a method of identifying glycopeptides in a sample, the method including, analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides, identifying the glycopeptides in the sample based on the at least one identified portion; and analyzing at least one glycopeptide of the identified glycopeptides.1. At least one computer-readable storage medium storing computer-executable instructions that, when executed by at least one processor, perform a method of identifying glycopeptides in a sample, the method comprising:
analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides; and identifying the glycopeptides in the sample based on the at least one identified portion. 2. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
determining the at least one characteristic of mass spectra indicative of presence of glycopeptides. 3. The at least one computer-readable storage medium of claim 2, wherein:
determining the at least one characteristic comprises determining at least one glycopeptide-rich acquisition enhancement zone. 4. The at least one computer-readable storage medium of claim 2, wherein:
determining the at least one characteristic comprises analyzing a training data set comprising a plurality of mass spectra of peptides. 5. The at least one computer-readable storage medium of claim 1, wherein:
the at least one characteristic comprises at least one first range of a nominal mass and at least one second range of mass defect. 6. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
displaying on a user interface results of the identification of the glycopeptides in the sample. 7. The at least one computer-readable storage medium of claim 6, wherein:
displaying the results of the identification of the glycopeptides comprises displaying the results so that the glycopeptides in the sample are differentiated from peptides in the sample. 8. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
providing a representation of the results of the identification of the glycopeptides so that the representation is enabled to receive input indicating selection of at least one glycopeptide of the identified glycopeptides for further analysis. 9. The at least one computer-readable storage medium of claim 8, wherein the method further comprises:
further analyzing the at least one glycopeptide selected for the further analysis. 10. The at least one computer-readable storage medium of claim 1, wherein:
identifying the glycopeptides in the sample comprises identifying N-glycosylated glycopeptides. 11. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
providing results of the identification of the glycopeptides in the sample to a system configured to further analyze the identified glycopeptides. 12. The at least one computer-readable storage medium of claim 1, wherein the method further comprises:
further analyzing at least one of the identified glycopeptides. 13. The at least one computer-readable storage medium of claim 1, wherein the sample comprises a biological sample. 14. The at least one computer-readable storage medium of claim 13, wherein the biological sample is obtained from tissue, urine, blood, plasma, serum or saliva. 15. The at least one computer-readable storage medium of claim 2, wherein:
the at least one characteristic is determined for a protease used to generate a mixture of peptides and glycopeptides from the sample. 16. The at least one computer-readable storage medium of claim 1, wherein:
analyzing the mass spectrum comprises analyzing precursor ion data. 17. At least one computer-readable storage medium storing computer-executable instructions that, when executed by at least one processor, perform a method of identifying glycopeptides in a sample, the method comprising:
determining at least one characteristic of mass spectra indicative of presence of glycopeptides; analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having the at least one characteristic; and identifying the glycopeptides in the sample based on the at least one identified portion. 18. A computer-implemented method of identifying glycopeptides in a sample, the method comprising:
with at least one processor:
analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides; and
identifying the glycopeptides in the sample based on the at least one identified portion. 19. The method of claim 18, further comprising:
determining the at least one characteristic of mass spectra indicative of presence of glycopeptides. 20. The method of claim 19, wherein:
determining the at least one characteristic comprises determining at least one glycopeptide-rich acquisition enhancement zone. 21. The method of claim 19, wherein:
determining the at least one characteristic comprises analyzing a data set comprising a plurality of mass spectra of peptides to determine at least one first range of a nominal mass and at least one second range of mass defect indicative of presence of glycopeptides. 22. The method of claim 19, wherein:
determining the at least one characteristic comprises analyzing a training data set comprising a plurality of mass spectra of peptides. 23. The method of claim 18, wherein:
the at least one characteristic comprises at least one first range of a nominal mass and at least one second range of mass defect. 24. The method of claim 18, further comprising:
displaying on a user interface results of the identification of the glycopeptides in the sample. 25. The method of claim 24, wherein:
displaying the results of the identification of the glycopeptides comprises displaying the results so that the glycopeptides in the sample are differentiated from peptides in the sample. 26. The method of claim 18, further comprising:
providing a representation of the results of the identification of the glycopeptides so that the representation is enabled to receive input indicating selection of at least one glycopeptide of the identified glycopeptides for further analysis. 27. The method of claim 26, further comprising:
further analyzing the at least one glycopeptide selected for the further analysis. 28. The method of claim 27, wherein:
further analyzing the at least one glycopeptide selected for the further analysis comprises determining a site of glycosylation on the at least one glycopeptide. 29. The system of claim 28, wherein:
determining the site of glycosylation comprises determining a site of N-glycosylation on the at least one glycopeptide. 30. The method of claim 27, further comprising:
analyzing the at least one glycopeptide using tandem mass-spectrometry. 31. The method of claim 18, wherein:
identifying the glycopeptides in the sample comprises identifying N-glycosylated glycopeptides. 32. The method of claim 18, further comprising:
providing results of the identification of the glycopeptides in the sample to a system configured to further analyze the identified glycopeptides. 33. The method of claim 18, further comprising:
further analyzing at least one of the identified glycopeptides. 34. The method of claim 18, wherein the sample comprises a biological sample. 35. The method of claim 34, wherein the biological sample is obtained from tissue, urine, blood, plasma, serum or saliva. 36. The method of claim 18, wherein:
analyzing the mass spectrum comprises analyzing precursor ion data. 37. A device comprising at least one processor and memory storing computer-executable instructions that, when executed by the at least one processor, perform a method of identifying glycopeptides in a sample, the method comprising:
analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides; and identifying the glycopeptides in the sample based on the at least one identified portion. 38. The device of claim 37, wherein the method further comprises:
determining the at least one characteristic of mass spectra indicative of presence of glycopeptides. 39. The device of claim 38, wherein:
determining the at least one characteristic comprises determining at least one glycopeptide-rich acquisition enhancement zone. 40. The device of claim 38, wherein:
determining the at least one characteristic comprises analyzing a data set comprising a plurality of mass spectra of peptides to determine at least one first range of a nominal mass and at least one second range of mass defect indicative of presence of glycopeptides. 41. The device of claim 38, wherein:
determining the at least one characteristic comprises analyzing a training data set comprising a plurality of mass spectra of peptides. 42. A system comprising:
a device comprising at least one processor and memory storing computer-executable instructions that, when executed by the at least one processor, perform a method of identifying glycopeptides in a sample, the method comprising:
analyzing a mass spectrum of the sample to identify at least one portion of the mass spectrum having at least one characteristic of mass spectra indicative of presence of glycopeptides;
identifying the glycopeptides in the sample based on the at least one identified portion; and
analyzing at least one glycopeptide of the identified glycopeptides. 43. The system of claim 42, wherein the method further comprises:
determining the at least one characteristic of mass spectra indicative of presence of glycopeptides. 44. The system of claim 43, wherein:
determining the at least one characteristic comprises determining at least one glycopeptide-rich acquisition enhancement zone. 45. The system of claim 43, wherein:
determining the at least one characteristic comprises analyzing a data set comprising a plurality of mass spectra of peptides to determine at least one first range of a nominal mass and at least one second range of mass defect indicative of presence of glycopeptides. 46. The system of claim 43, wherein:
determining the at least one characteristic comprises analyzing a training data set comprising a plurality of mass spectra of peptides. 47. The system of claim 42, wherein:
analyzing the at least one glycopeptide comprises determining a site of glycosylation on the at least one glycopeptide. 48. The system of claim 47, wherein:
determining the site of glycosylation comprises determining a site of N-glycosylation on the at least one glycopeptide. | 1,600 |
1,094 | 14,772,063 | 1,634 | Provided herein is a method for making a pool of probes by primer extension. In certain embodiments, the method comprises hybridizing a first population of oligonucleotides comprising a top strand sequence having the following formula V1-B-3′ with a second population of oligonucleotides comprising a bottom strand sequence having the following formula V2′-B′-3′ to provide a population of duplexes. After hybridizing, the 3′ ends of the oligonucleotides in the duplexes are extended to produce a population of double stranded products comprising a top strand sequence having the following formula V1-B-V2, where V2 is complementary to V2′. | 1. A method comprising:
a) hybridizing a first population of oligonucleotides comprising a top strand sequence having the following formula:
V1-B-3′
with a second population of oligonucleotides comprising a bottom strand sequence having the following formula:
V2′-B′-3′
to provide a population of duplexes; wherein:
the nucleotide sequences of B and B′ are complementary and are at least 15 nucleotides in length;
the nucleotide sequence of B is the same for each oligonucleotide of said first population;
the nucleotide sequence of B′ is the same for each oligonucleotide of said second population;
the nucleotide sequence of V1 is variable between the oligonucleotides of the first population
the nucleotide sequence of V2′ is variable between the oligonucleotides of the second population, and
V1 and V2′ hybridize to sites in a reference genome; and b) extending the 3′ ends of the oligonucleotides in said duplexes to produce a population of double stranded products comprising a top strand sequence having the following formula:
V1-B-V2,
wherein V2 is complementary to V2′. 2. The method of claim 1, wherein:
said first population of oligonucleotides comprises a top strand sequence having the following formula:
F-V1-B-3′;
said second population of oligonucleotides comprises a bottom strand sequence having the following formula:
R′-V2′-B′-3′;
and said population of double-stranded products comprise a top strand sequence having the following formula:
F-V1-B-V2-R
wherein R and the complement of F provide binding sites for forward and reverse primers that can be used to amplify said population of double-stranded products to produce a population of PCR products. 3. The method of claim 2, further comprising:
c) PCR amplifying said population of double-stranded products using said forward and reverse primers to produce said population of PCR products. 4. The method of claim 4, further comprising:
d) isolating said top strand from the bottom strand of said population of PCR products to produce a population of single stranded halo probes that hybridize to a plurality of sites in said genome. 5. The method of claim 4, wherein said reverse primer is 5′-phosphorylated and said isolating is done by degrading the bottom strand of said population of double-stranded products using an exonuclease. 6. The method of claim 5, wherein said exonuclease is λ exonuclease. 7. The method of claim 1, wherein said genome is a mammalian genome. 8. The method of claim 1, wherein the nucleotide sequence of B is at least 15 bases in length. 9. The method of claim 1, wherein the nucleotide sequences of V1 and V2′ are at least 25 nucleotides in length 10. The method of claim 1, wherein said first population of oligonucleotides comprises at least 10 oligonucleotides and said second population of oligonucleotides comprises at least 10 oligonucleotides. 11. The method of claim 1, wherein the sites to which V1 and V2 hybridize in said genome each comprise a restriction site. 12. A composition of matter comprising:
a population of duplexes, wherein said duplexes comprise: a first population of oligonucleotides comprising a top strand sequence having the following formula:
V1-B-3′; and
a second population of oligonucleotides comprising a bottom strand sequence having the following formula:
V2′-B′-3′; wherein:
the nucleotide sequences of B and B′ are complementary and are at least 15 nucleotides in length;
the nucleotide sequence of B is the same for each oligonucleotide of said first population;
the nucleotide sequence of B′ is the same for each oligonucleotide of said second population;
the nucleotide sequence of V1 is variable between the oligonucleotides of the first population
the nucleotide sequence of V2′ is variable between the oligonucleotides of the second population, and
V1 and V2′ hybridize to different sites in a reference genome. 13. The composition of matter of claim 12, wherein the nucleotide sequence of B is at least 15 bases in length. 14. The composition of matter of claim 12, wherein the nucleotide sequences of V1 and V2′ are at least 25 nucleotides in length 15. The composition of matter of claim 12, wherein said first population of oligonucleotides comprises at least 10 oligonucleotides and said second population of oligonucleotides comprises at least 10 oligonucleotides. 16. A kit comprising
a) a first population of oligonucleotides comprising a top strand sequence having the following formula:
V1-B-3′; and
b) a second population of oligonucleotides comprising a bottom strand sequence having the following formula:
V2′-B′-3′; wherein:
the nucleotide sequences of B and B′ are complementary and are at least 15 nucleotides in length;
the nucleotide sequence of B is the same for each oligonucleotide of said first population;
the nucleotide sequence of B′ is the same for each oligonucleotide of said second population;
the nucleotide sequence of V1 is variable between the oligonucleotides of the first population
the nucleotide sequence of V2′ is variable between the oligonucleotides of the second population, and
V1 and V2′ hybridize to different sites in a reference genome. 17. The kit of claim 16, further comprising instructions for performing the method of claim 1. 18. The kit of claim 16, wherein the first population of oligonucleotides comprises a top strand sequence having the following formula:
F-V1-B-3′; and
and the second population of oligonucleotides comprises a bottom strand sequence having the following formula:
R′-V2′-B′-3′;
wherein the complements of sequences F and R′ provide binding sites for forward and reverse primers. 19. The kit of claim 16, further comprising forward and reverse primers that hybridize to the complements of sequences F and R′. 20. The kit of claim 16, wherein one of said forward and reverse primers comprises a 5′ phosphate or biotin. | Provided herein is a method for making a pool of probes by primer extension. In certain embodiments, the method comprises hybridizing a first population of oligonucleotides comprising a top strand sequence having the following formula V1-B-3′ with a second population of oligonucleotides comprising a bottom strand sequence having the following formula V2′-B′-3′ to provide a population of duplexes. After hybridizing, the 3′ ends of the oligonucleotides in the duplexes are extended to produce a population of double stranded products comprising a top strand sequence having the following formula V1-B-V2, where V2 is complementary to V2′.1. A method comprising:
a) hybridizing a first population of oligonucleotides comprising a top strand sequence having the following formula:
V1-B-3′
with a second population of oligonucleotides comprising a bottom strand sequence having the following formula:
V2′-B′-3′
to provide a population of duplexes; wherein:
the nucleotide sequences of B and B′ are complementary and are at least 15 nucleotides in length;
the nucleotide sequence of B is the same for each oligonucleotide of said first population;
the nucleotide sequence of B′ is the same for each oligonucleotide of said second population;
the nucleotide sequence of V1 is variable between the oligonucleotides of the first population
the nucleotide sequence of V2′ is variable between the oligonucleotides of the second population, and
V1 and V2′ hybridize to sites in a reference genome; and b) extending the 3′ ends of the oligonucleotides in said duplexes to produce a population of double stranded products comprising a top strand sequence having the following formula:
V1-B-V2,
wherein V2 is complementary to V2′. 2. The method of claim 1, wherein:
said first population of oligonucleotides comprises a top strand sequence having the following formula:
F-V1-B-3′;
said second population of oligonucleotides comprises a bottom strand sequence having the following formula:
R′-V2′-B′-3′;
and said population of double-stranded products comprise a top strand sequence having the following formula:
F-V1-B-V2-R
wherein R and the complement of F provide binding sites for forward and reverse primers that can be used to amplify said population of double-stranded products to produce a population of PCR products. 3. The method of claim 2, further comprising:
c) PCR amplifying said population of double-stranded products using said forward and reverse primers to produce said population of PCR products. 4. The method of claim 4, further comprising:
d) isolating said top strand from the bottom strand of said population of PCR products to produce a population of single stranded halo probes that hybridize to a plurality of sites in said genome. 5. The method of claim 4, wherein said reverse primer is 5′-phosphorylated and said isolating is done by degrading the bottom strand of said population of double-stranded products using an exonuclease. 6. The method of claim 5, wherein said exonuclease is λ exonuclease. 7. The method of claim 1, wherein said genome is a mammalian genome. 8. The method of claim 1, wherein the nucleotide sequence of B is at least 15 bases in length. 9. The method of claim 1, wherein the nucleotide sequences of V1 and V2′ are at least 25 nucleotides in length 10. The method of claim 1, wherein said first population of oligonucleotides comprises at least 10 oligonucleotides and said second population of oligonucleotides comprises at least 10 oligonucleotides. 11. The method of claim 1, wherein the sites to which V1 and V2 hybridize in said genome each comprise a restriction site. 12. A composition of matter comprising:
a population of duplexes, wherein said duplexes comprise: a first population of oligonucleotides comprising a top strand sequence having the following formula:
V1-B-3′; and
a second population of oligonucleotides comprising a bottom strand sequence having the following formula:
V2′-B′-3′; wherein:
the nucleotide sequences of B and B′ are complementary and are at least 15 nucleotides in length;
the nucleotide sequence of B is the same for each oligonucleotide of said first population;
the nucleotide sequence of B′ is the same for each oligonucleotide of said second population;
the nucleotide sequence of V1 is variable between the oligonucleotides of the first population
the nucleotide sequence of V2′ is variable between the oligonucleotides of the second population, and
V1 and V2′ hybridize to different sites in a reference genome. 13. The composition of matter of claim 12, wherein the nucleotide sequence of B is at least 15 bases in length. 14. The composition of matter of claim 12, wherein the nucleotide sequences of V1 and V2′ are at least 25 nucleotides in length 15. The composition of matter of claim 12, wherein said first population of oligonucleotides comprises at least 10 oligonucleotides and said second population of oligonucleotides comprises at least 10 oligonucleotides. 16. A kit comprising
a) a first population of oligonucleotides comprising a top strand sequence having the following formula:
V1-B-3′; and
b) a second population of oligonucleotides comprising a bottom strand sequence having the following formula:
V2′-B′-3′; wherein:
the nucleotide sequences of B and B′ are complementary and are at least 15 nucleotides in length;
the nucleotide sequence of B is the same for each oligonucleotide of said first population;
the nucleotide sequence of B′ is the same for each oligonucleotide of said second population;
the nucleotide sequence of V1 is variable between the oligonucleotides of the first population
the nucleotide sequence of V2′ is variable between the oligonucleotides of the second population, and
V1 and V2′ hybridize to different sites in a reference genome. 17. The kit of claim 16, further comprising instructions for performing the method of claim 1. 18. The kit of claim 16, wherein the first population of oligonucleotides comprises a top strand sequence having the following formula:
F-V1-B-3′; and
and the second population of oligonucleotides comprises a bottom strand sequence having the following formula:
R′-V2′-B′-3′;
wherein the complements of sequences F and R′ provide binding sites for forward and reverse primers. 19. The kit of claim 16, further comprising forward and reverse primers that hybridize to the complements of sequences F and R′. 20. The kit of claim 16, wherein one of said forward and reverse primers comprises a 5′ phosphate or biotin. | 1,600 |
1,095 | 15,436,984 | 1,617 | The present invention is directed to novel insecticidal and/or miticidal compositions comprising bifenthrin and a cyano-pyrethroid. The compositions exhibit an unexpected increase in insecticidal activity as compared to the insecticidal activity of the individual components. | 1. A miticidal composition comprising bifenthrin and a cyano-pyrethroid selected from the group consisting of acrinathrin, cycloprothrin, tralomethrin, fenvalerate, beta-cyfluthrin, flucythrinate, beta-cypermethrin, theta-cypermethrin, cyphenothrin, cyhalothrin, fluvalinate and fenpropathrin. 2. A composition of claim 1 where the ratio of bifenthrin to cyano-pyrethroid is from 1/99 to 99/1. 3. A composition of claim 1 wherein the cyano-pyrethroid is selected from the group consisting of deltamethrin, cyfluthrin, alpha-cypermethrin, zeta-cypermethrin, lambda-cyhalothrin and esfenvalerate. 4. A composition of claim 1 wherein the cyano-pyrethroid is zeta-cypermethrin. 5. A composition of claim 4 wherein the zeta-cypermethrin is (R,S)-α-cyano-3-phenoxybenzyl-(1RS)-cis-trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylate which has been enriched in the 1R-cis-S and 1R-trans-S isomers by the reaction of a 55/45 cis/trans mixture of cypermethrin with a catalytic amount of tricaprylylammonium chloride and sodium carbonate in n-heptane. 6. A composition of claim 1 further comprising an agriculturally acceptable extender or adjuvant. 7. A method for controlling unwanted insects or mites comprising applying a composition of claim 1 to a locus where insects or mites are present or are expected to be present. | The present invention is directed to novel insecticidal and/or miticidal compositions comprising bifenthrin and a cyano-pyrethroid. The compositions exhibit an unexpected increase in insecticidal activity as compared to the insecticidal activity of the individual components.1. A miticidal composition comprising bifenthrin and a cyano-pyrethroid selected from the group consisting of acrinathrin, cycloprothrin, tralomethrin, fenvalerate, beta-cyfluthrin, flucythrinate, beta-cypermethrin, theta-cypermethrin, cyphenothrin, cyhalothrin, fluvalinate and fenpropathrin. 2. A composition of claim 1 where the ratio of bifenthrin to cyano-pyrethroid is from 1/99 to 99/1. 3. A composition of claim 1 wherein the cyano-pyrethroid is selected from the group consisting of deltamethrin, cyfluthrin, alpha-cypermethrin, zeta-cypermethrin, lambda-cyhalothrin and esfenvalerate. 4. A composition of claim 1 wherein the cyano-pyrethroid is zeta-cypermethrin. 5. A composition of claim 4 wherein the zeta-cypermethrin is (R,S)-α-cyano-3-phenoxybenzyl-(1RS)-cis-trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylate which has been enriched in the 1R-cis-S and 1R-trans-S isomers by the reaction of a 55/45 cis/trans mixture of cypermethrin with a catalytic amount of tricaprylylammonium chloride and sodium carbonate in n-heptane. 6. A composition of claim 1 further comprising an agriculturally acceptable extender or adjuvant. 7. A method for controlling unwanted insects or mites comprising applying a composition of claim 1 to a locus where insects or mites are present or are expected to be present. | 1,600 |
1,096 | 15,697,765 | 1,629 | An aqueous solution includes a silver complex and a pH buffer. The silver complex may include a silver atom in complex with an organic acid compound, such as dihydrogen citrate. | 1. A biocide comprising:
an aqueous solution including a silver complex and a pH buffer. 2. The biocide as recited in claim 1, wherein the aqueous solution has a pH of 6-10, and a pH of 3 or less without the pH buffer. 3. The biocide as recited in claim 1, wherein the silver complex includes a silver atom in complex with an organic acid compound. 4. The biocide as recited in claim 1, wherein the silver complex includes a silver atom in complex with a citrate compound. 5. The biocide as recited in claim 1, wherein the silver complex is silver dihydrogen citrate. 6. The biocide as recited in claim 1, wherein the aqueous solution has a silver concentration of up to 30 parts-per-million. 7. The biocide as recited in claim 1, wherein the pH buffer is selected from the group consisting of carbonate, phosphate, glycine, hydroxide, and combinations thereof. 8. The biocide as recited in claim 1, wherein the pH buffer includes sodium carbonate and sodium bicarbonate. 9. The biocide as recited in claim 8, wherein the silver complex is silver dihydrogen citrate, and the aqueous solution has a silver concentration of up to 30 parts-per-million. 10. The biocide as recited in claim 1, wherein the pH buffer is selected from the group consisting of tris, tricine, 3-morpholinopropanesulfonic acid, and combinations thereof. 11. The biocide as recited in claim 1, wherein the pH buffer includes at least two buffers selected from the group consisting of tris, tricine, 3-morpholinopropanesulfonic acid, carbonate, phosphate, glycine, hydroxide, and combinations thereof. 12. A method for producing a biocide, the method comprising:
for an aqueous solution that has a silver complex and a pH that is below 6, increasing the pH of the aqueous solution into a pH range of 6-10 by adding a pH buffer to the aqueous solution. 13. The method as recited in claim 12, wherein the pH of the aqueous solution prior to adding the pH buffer is 3 or less. 14. The method as recited in claim 12, wherein the silver complex includes a silver atom in complex with an organic acid compound. 15. The method as recited in claim 12, wherein the silver complex includes a silver atom in complex with a citrate compound. 16. The method as recited in claim 12, wherein the aqueous solution after adding the pH buffer has a silver concentration of up to 30 parts-per-million. 17. The method as recited in claim 12, wherein the pH buffer is selected from the group consisting of carbonate, phosphate, glycine, hydroxide, and combinations thereof. 18. The method as recited in claim 12, wherein the pH buffer includes at least one of sodium carbonate or sodium bicarbonate. 19. The method as recited in claim 12, wherein the pH buffer is selected from the group consisting of tris, tricine, 3-morpholinopropanesulfonic acid, and combinations thereof. 20. The method as recited in claim 12, wherein adding the pH buffer includes adding at least two buffers selected from the group consisting of tris, tricine, 3-morpholinopropanesulfonic acid, carbonate, phosphate, glycine, hydroxide, and combinations thereof. | An aqueous solution includes a silver complex and a pH buffer. The silver complex may include a silver atom in complex with an organic acid compound, such as dihydrogen citrate.1. A biocide comprising:
an aqueous solution including a silver complex and a pH buffer. 2. The biocide as recited in claim 1, wherein the aqueous solution has a pH of 6-10, and a pH of 3 or less without the pH buffer. 3. The biocide as recited in claim 1, wherein the silver complex includes a silver atom in complex with an organic acid compound. 4. The biocide as recited in claim 1, wherein the silver complex includes a silver atom in complex with a citrate compound. 5. The biocide as recited in claim 1, wherein the silver complex is silver dihydrogen citrate. 6. The biocide as recited in claim 1, wherein the aqueous solution has a silver concentration of up to 30 parts-per-million. 7. The biocide as recited in claim 1, wherein the pH buffer is selected from the group consisting of carbonate, phosphate, glycine, hydroxide, and combinations thereof. 8. The biocide as recited in claim 1, wherein the pH buffer includes sodium carbonate and sodium bicarbonate. 9. The biocide as recited in claim 8, wherein the silver complex is silver dihydrogen citrate, and the aqueous solution has a silver concentration of up to 30 parts-per-million. 10. The biocide as recited in claim 1, wherein the pH buffer is selected from the group consisting of tris, tricine, 3-morpholinopropanesulfonic acid, and combinations thereof. 11. The biocide as recited in claim 1, wherein the pH buffer includes at least two buffers selected from the group consisting of tris, tricine, 3-morpholinopropanesulfonic acid, carbonate, phosphate, glycine, hydroxide, and combinations thereof. 12. A method for producing a biocide, the method comprising:
for an aqueous solution that has a silver complex and a pH that is below 6, increasing the pH of the aqueous solution into a pH range of 6-10 by adding a pH buffer to the aqueous solution. 13. The method as recited in claim 12, wherein the pH of the aqueous solution prior to adding the pH buffer is 3 or less. 14. The method as recited in claim 12, wherein the silver complex includes a silver atom in complex with an organic acid compound. 15. The method as recited in claim 12, wherein the silver complex includes a silver atom in complex with a citrate compound. 16. The method as recited in claim 12, wherein the aqueous solution after adding the pH buffer has a silver concentration of up to 30 parts-per-million. 17. The method as recited in claim 12, wherein the pH buffer is selected from the group consisting of carbonate, phosphate, glycine, hydroxide, and combinations thereof. 18. The method as recited in claim 12, wherein the pH buffer includes at least one of sodium carbonate or sodium bicarbonate. 19. The method as recited in claim 12, wherein the pH buffer is selected from the group consisting of tris, tricine, 3-morpholinopropanesulfonic acid, and combinations thereof. 20. The method as recited in claim 12, wherein adding the pH buffer includes adding at least two buffers selected from the group consisting of tris, tricine, 3-morpholinopropanesulfonic acid, carbonate, phosphate, glycine, hydroxide, and combinations thereof. | 1,600 |
1,097 | 14,654,650 | 1,618 | There is provided a microparticle composition suitable for molecular imaging, the composition comprising microparticles, wherein the microparticles comprise: a core microparticle structure having a central area and a shell, and wherein the core microparticle structure comprises (i) a phosphatidylcholine lipid: (ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and (iii) an alkoxylated fatty acid. | 1. A microparticle composition suitable for molecular imaging, the composition comprising microparticles, wherein the microparticles comprise: a core microparticle structure having a central area and a shell, and wherein the core microparticle structure comprises:
(i) a phosphatidylcholine lipid; (ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and (iii) an alkoxylated fatty acid. 2. A microparticle composition according to claim 1, further comprising at least one molecular binding element, wherein the at least one molecular binding element is covalently attached to the shell of the core microparticle structure. 3. A microparticle composition according to claim 2, wherein the at least one molecular binding element is covalently attached to the core microparticle structure via the at least one maleimide moiety. 4. A microparticle composition according to claim 1, wherein the core microparticle structure is micellar. 5. A microparticle composition according to claim 1, wherein the molar ratio of (i) to (iii) satisfies the following ranges:
(iv) 70 to 80; (v) 5 to 15; and (vi) 10 to 20. 6. A microparticle composition according to claim 1, wherein the core microparticle structure further comprises a labelling moiety. 7. A microparticle composition according to claim 6, wherein the molar ratio of the labelling moiety in the composition is 0.2 to 50. 8. A microparticle composition according to claim 7, wherein the molar ratio of the labelling moiety in the composition is 0.5 to 5. 9. A microparticle composition according to claim 6, wherein the labelling moiety is a fluorescent dye. 10. A microparticle composition according to claim 9, wherein the labelling moiety is 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI). 11. A microparticle composition according to claim 1, wherein the core microparticle structure further comprises a phosphatidylethanolamine lipid. 12. A microparticle composition according to claim 11, wherein the phosphatidylethanolamine lipid is a C10-20 saturated phosphatidylethanolamine lipid. 13. A microparticle composition according to claim 12, wherein the phosphatidylethanolamine lipid is a distearoylphosphatidylethanolamine (DSPE). 14. A microparticle composition according to claim 11, wherein the molar ratio of the phosphatidylethanolamine lipid in the composition is 0.5 to 5. 15. A microparticle composition according to claim 1, wherein the central area of the core microparticle structure contains a fluid medium. 16. A microparticle composition according to claim 15, wherein the fluid medium comprises a physiologically acceptable gas. 17. A microparticle composition according to claim 16, wherein the physiologically acceptable gas is selected from air, nitrogen, carbon dioxide, xenon, krypton, sulfur hexafluoride, chlorotrifluoromethane, dichlorodifluoro-methane, bromotrifluoromethane, bromochlorodifluoromethane, tetrafluoromethane, dibromo-difluoromethane, dichlorotetrafluoroethane, chloropentafluoroethane, hexafluoroethane, hexafluoropropylene, octafluoropropane, hexafluoro-butadiene, octafluoro-2-butene, octafluorocyclobutane, decafluorobutane, perfluorocyclopentane, dodecafluoropentane, and tetradecafluorohexane. 18. A microparticle composition according to claim 16, wherein the physiologically acceptable gas comprises octafluoropropane. 19. A microparticle composition according to claim 2, wherein the at least one molecular binding element comprises a protein, peptide, or small organic moiety. 20. A microparticle composition according to claim 19, wherein the at least one molecular binding element is an antibody. 21. A microparticle composition according to claim 2, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) is ≧1:1. 22. A microparticle composition according to claim 21, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) is ≧5:1. 23. A microparticle composition according to claim 2, wherein the microparticles have at least about 1×105 molecular binding elements per microparticle. 24. A microparticle composition according to claim 1, wherein the phosphatidylcholine lipid is a C10-20 saturated phosphatidylcholine lipid. 25. A microparticle composition according to claim 24, wherein the phosphatidylcholine lipid is a 1,2-distearoyl-sn-glycero-3-phosphocholine. 26. A microparticle composition according to claim 1, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety is a C10-20 saturated phosphatidylethanolamine lipid. 27. A microparticle composition according to claim 1, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety comprises a polyethylene glycol chain with a molecular weight of at least 500. 28. A microparticle composition according to claim 27, wherein the at least one maleimide moiety is attached to the polyethylene glycol chain. 29. A microparticle composition according to claim 28, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety is a 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000]. 30. A microparticle composition according to claim 1, wherein the alkoxylated fatty acid is a polyethylene glycol fatty acid ester. 31. A microparticle composition according to claim 30, wherein the polyethylene glycol fatty acid ester is a C10-20 saturated polyethylene glycol fatty acid ester. 32. A microparticle composition according to claim 31, wherein the polyethylene glycol fatty acid ester is a PEG40 stearate. 33. A microparticle composition according to claim 1, wherein the average diameter of the microparticles is in the range of 0.5 to 5 μm. 34. An intermediate microparticle composition suitable for producing a molecular imaging microparticle composition, wherein the intermediate composition comprises:
(i) a phosphatidylcholine lipid; (ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and (iii) an alkoxylated fatty acid. 35. An intermediate microparticle composition according to claim 34, further comprising at least one molecular binding element. 36. An intermediate microparticle composition according to claim 35, wherein the at least one molecular binding element is covalently attached to the at least one maleimide moiety. 37. An intermediate microparticle composition according to claim 34, wherein the molar ratio of (i) to (iii) satisfies the following ranges:
(i) 70 to 80; (ii) 5 to 15; and (iii) 10 to 20. 38. An intermediate microparticle composition according to claim 34, further comprising a labelling moiety. 39. An intermediate microparticle composition according to claim 38, wherein the molar ratio of the labelling moiety in the composition is 0.2 to 50. 40. An intermediate microparticle composition according to claim 39, wherein the molar ratio of the labelling moiety in the composition is 0.5 to 5. 41. An intermediate microparticle composition according to claim 38, wherein the labelling moiety is a fluorescent dye. 42. An intermediate microparticle composition according to claim 41, wherein the labelling moiety is a 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI). 43. An intermediate microparticle composition according to claim 38, further comprising a phosphatidylethanolamine lipid. 44. An intermediate microparticle composition according to claim 43, wherein the phosphatidylethanolamine lipid is a C10-20 saturated phosphatidylethanolamine lipid. 45. An intermediate microparticle composition according to claim 44, wherein the phosphatidylethanolamine lipid is a distearoylphosphatidylethanolamine (DSPE). 46. An intermediate microparticle composition according to claim 43, wherein the molar ratio of the phosphatidylethanolamine lipid in the composition is 0.5 to 5. 47. An intermediate microparticle composition according to claim 35, wherein the at least one molecular binding element comprises a protein, peptide, or small organic moiety. 48. An intermediate microparticle composition according to claim 47, wherein the at least one molecular binding element is an antibody. 49. An intermediate microparticle composition according to claim 35, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) is ≧1:1. 50. An intermediate microparticle composition according to claim 49, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) is ≧5:1. 51. An intermediate microparticle composition according to claim 34, wherein the phosphatidylcholine lipid is a C10-20 saturated phosphatidylcholine lipid. 52. An intermediate microparticle composition according to claim 51, wherein the phosphatidylcholine lipid is a 1,2-distearoyl-sn-glycero-3-phosphocholine. 53. An intermediate microparticle composition according to claim 34, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety is a C10-20 saturated phosphatidylethanolamine lipid. 54. An intermediate microparticle composition according to any of claims 34, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety comprises a polyethylene glycol chain with a molecular weight of at least 500. 55. An intermediate microparticle composition according to claim 54, wherein the at least one maleimide moiety is attached to the polyethylene glycol chain. 56. An intermediate microparticle composition according to claim 55, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety is a 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000]. 57. An intermediate microparticle composition according to claim 34, wherein the alkoxylated fatty acid is a polyethylene glycol fatty acid ester. 58. An intermediate microparticle composition according to claim 57, wherein the polyethylene glycol fatty acid ester is a C10-20 saturated polyethylene glycol fatty acid ester. 59. An intermediate microparticle composition according to claim 58, wherein the polyethylene glycol fatty acid ester is a PEG40 stearate. 60. An intermediate microparticle composition according to claim 34, wherein the composition is a lyophilisate. 61. An intermediate microparticle composition according to claim 34, wherein the composition is an aqueous dispersion 62. A kit comprising an intermediate composition according to claim 34, and a source of a fluid medium which comprises a physiologically acceptable gas. 63. A kit according to claim 62, wherein the physiologically acceptable gas is selected from air, nitrogen, carbon dioxide, xenon, krypton, sulfur hexafluoride, chlorotrifluoromethane, dichlorodifluoro-methane, bromotrifluoromethane, bromochlorodifluoromethane, tetrafluoromethane, dibromo-difluoromethane, dichlorotetrafluoroethane, chloropentafluoroethane, hexafluoroethane, hexafluoropropylene, octafluoropropane, hexafluoro-butadiene, octafluoro-2-butene, octafluorocyclobutane, decafluorobutane, perfluorocyclopentane, dodecafluoropentane, and tetradecafluorohexane. 64. A kit according to claim 63, wherein the physiologically acceptable gas comprises octafluoropropane. 65. A method of preparing a microparticle composition, the method comprising:
forming a core microparticle structure having a central area and a shell comprising
(i) a phosphatidylcholine lipid;
(ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and
(iii) an alkoxylated fatty acid. 66. A method of preparing a microparticle composition according to claim 65, the method further comprising:
(ii) covalently attaching at least one molecular binding element to the shell of the core microparticle structure. 67. A method of preparing a microparticle composition according to claim 65, wherein step (i) is conducted in the presence of a physiologically acceptable gas. 68. A method of preparing a microparticle composition according to claim 66, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) in step (i) is ≧1:1. 69. A method of preparing a microparticle composition according to claim 68, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) in step (i) is ≧5:1. 70. A pharmaceutical composition comprising a microparticle composition according to claim 1, and a pharmaceutically acceptable carrier or excipient. 71. A method of molecular imaging comprising:
providing a pharmaceutical composition, the pharmaceutical composition comprising a microparticle composition and a at least one of a pharmaceutically acceptable carrier and excipient, the pharmaceutical composition including a contrast agent, the contrast agent including: i. a phosphatidylcholine lipid; ii. a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and iii. an alkoxylated fatty acid 72. A method of molecular imaging comprising:
providing a microparticle composition, the microparticle composition including a contrast agent, the contrast agent including: iv. a phosphatidylcholine lipid; v. a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and vi. an alkoxylated fatty acid 73. An imaging method comprising:
administering at least one of a microparticle composition and a pharmaceutical composition comprising:
(i) a phosphatidylcholine lipid;
(ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and
(iii) an alkoxylated fatty acid;
and
imaging the subject using at least one imaging technique. 74. An imaging method according to claim 73, wherein the imaging technique comprises ultrasound imaging (US). 75. An imaging method according to claim 73, wherein the imaging technique comprises magnetic resonance imaging (MRI). 76. A microparticle composition suitable for molecular imaging, the composition comprising microparticles, wherein the microparticles comprise: a core microparticle structure having a central area and a shell, and wherein the core microparticle structure comprises:
(i) a phosphatidylcholine lipid; (ii) a phosphatidylethanolamine lipid; and (iii) an alkoxylated fatty acid. | There is provided a microparticle composition suitable for molecular imaging, the composition comprising microparticles, wherein the microparticles comprise: a core microparticle structure having a central area and a shell, and wherein the core microparticle structure comprises (i) a phosphatidylcholine lipid: (ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and (iii) an alkoxylated fatty acid.1. A microparticle composition suitable for molecular imaging, the composition comprising microparticles, wherein the microparticles comprise: a core microparticle structure having a central area and a shell, and wherein the core microparticle structure comprises:
(i) a phosphatidylcholine lipid; (ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and (iii) an alkoxylated fatty acid. 2. A microparticle composition according to claim 1, further comprising at least one molecular binding element, wherein the at least one molecular binding element is covalently attached to the shell of the core microparticle structure. 3. A microparticle composition according to claim 2, wherein the at least one molecular binding element is covalently attached to the core microparticle structure via the at least one maleimide moiety. 4. A microparticle composition according to claim 1, wherein the core microparticle structure is micellar. 5. A microparticle composition according to claim 1, wherein the molar ratio of (i) to (iii) satisfies the following ranges:
(iv) 70 to 80; (v) 5 to 15; and (vi) 10 to 20. 6. A microparticle composition according to claim 1, wherein the core microparticle structure further comprises a labelling moiety. 7. A microparticle composition according to claim 6, wherein the molar ratio of the labelling moiety in the composition is 0.2 to 50. 8. A microparticle composition according to claim 7, wherein the molar ratio of the labelling moiety in the composition is 0.5 to 5. 9. A microparticle composition according to claim 6, wherein the labelling moiety is a fluorescent dye. 10. A microparticle composition according to claim 9, wherein the labelling moiety is 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI). 11. A microparticle composition according to claim 1, wherein the core microparticle structure further comprises a phosphatidylethanolamine lipid. 12. A microparticle composition according to claim 11, wherein the phosphatidylethanolamine lipid is a C10-20 saturated phosphatidylethanolamine lipid. 13. A microparticle composition according to claim 12, wherein the phosphatidylethanolamine lipid is a distearoylphosphatidylethanolamine (DSPE). 14. A microparticle composition according to claim 11, wherein the molar ratio of the phosphatidylethanolamine lipid in the composition is 0.5 to 5. 15. A microparticle composition according to claim 1, wherein the central area of the core microparticle structure contains a fluid medium. 16. A microparticle composition according to claim 15, wherein the fluid medium comprises a physiologically acceptable gas. 17. A microparticle composition according to claim 16, wherein the physiologically acceptable gas is selected from air, nitrogen, carbon dioxide, xenon, krypton, sulfur hexafluoride, chlorotrifluoromethane, dichlorodifluoro-methane, bromotrifluoromethane, bromochlorodifluoromethane, tetrafluoromethane, dibromo-difluoromethane, dichlorotetrafluoroethane, chloropentafluoroethane, hexafluoroethane, hexafluoropropylene, octafluoropropane, hexafluoro-butadiene, octafluoro-2-butene, octafluorocyclobutane, decafluorobutane, perfluorocyclopentane, dodecafluoropentane, and tetradecafluorohexane. 18. A microparticle composition according to claim 16, wherein the physiologically acceptable gas comprises octafluoropropane. 19. A microparticle composition according to claim 2, wherein the at least one molecular binding element comprises a protein, peptide, or small organic moiety. 20. A microparticle composition according to claim 19, wherein the at least one molecular binding element is an antibody. 21. A microparticle composition according to claim 2, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) is ≧1:1. 22. A microparticle composition according to claim 21, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) is ≧5:1. 23. A microparticle composition according to claim 2, wherein the microparticles have at least about 1×105 molecular binding elements per microparticle. 24. A microparticle composition according to claim 1, wherein the phosphatidylcholine lipid is a C10-20 saturated phosphatidylcholine lipid. 25. A microparticle composition according to claim 24, wherein the phosphatidylcholine lipid is a 1,2-distearoyl-sn-glycero-3-phosphocholine. 26. A microparticle composition according to claim 1, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety is a C10-20 saturated phosphatidylethanolamine lipid. 27. A microparticle composition according to claim 1, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety comprises a polyethylene glycol chain with a molecular weight of at least 500. 28. A microparticle composition according to claim 27, wherein the at least one maleimide moiety is attached to the polyethylene glycol chain. 29. A microparticle composition according to claim 28, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety is a 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000]. 30. A microparticle composition according to claim 1, wherein the alkoxylated fatty acid is a polyethylene glycol fatty acid ester. 31. A microparticle composition according to claim 30, wherein the polyethylene glycol fatty acid ester is a C10-20 saturated polyethylene glycol fatty acid ester. 32. A microparticle composition according to claim 31, wherein the polyethylene glycol fatty acid ester is a PEG40 stearate. 33. A microparticle composition according to claim 1, wherein the average diameter of the microparticles is in the range of 0.5 to 5 μm. 34. An intermediate microparticle composition suitable for producing a molecular imaging microparticle composition, wherein the intermediate composition comprises:
(i) a phosphatidylcholine lipid; (ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and (iii) an alkoxylated fatty acid. 35. An intermediate microparticle composition according to claim 34, further comprising at least one molecular binding element. 36. An intermediate microparticle composition according to claim 35, wherein the at least one molecular binding element is covalently attached to the at least one maleimide moiety. 37. An intermediate microparticle composition according to claim 34, wherein the molar ratio of (i) to (iii) satisfies the following ranges:
(i) 70 to 80; (ii) 5 to 15; and (iii) 10 to 20. 38. An intermediate microparticle composition according to claim 34, further comprising a labelling moiety. 39. An intermediate microparticle composition according to claim 38, wherein the molar ratio of the labelling moiety in the composition is 0.2 to 50. 40. An intermediate microparticle composition according to claim 39, wherein the molar ratio of the labelling moiety in the composition is 0.5 to 5. 41. An intermediate microparticle composition according to claim 38, wherein the labelling moiety is a fluorescent dye. 42. An intermediate microparticle composition according to claim 41, wherein the labelling moiety is a 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI). 43. An intermediate microparticle composition according to claim 38, further comprising a phosphatidylethanolamine lipid. 44. An intermediate microparticle composition according to claim 43, wherein the phosphatidylethanolamine lipid is a C10-20 saturated phosphatidylethanolamine lipid. 45. An intermediate microparticle composition according to claim 44, wherein the phosphatidylethanolamine lipid is a distearoylphosphatidylethanolamine (DSPE). 46. An intermediate microparticle composition according to claim 43, wherein the molar ratio of the phosphatidylethanolamine lipid in the composition is 0.5 to 5. 47. An intermediate microparticle composition according to claim 35, wherein the at least one molecular binding element comprises a protein, peptide, or small organic moiety. 48. An intermediate microparticle composition according to claim 47, wherein the at least one molecular binding element is an antibody. 49. An intermediate microparticle composition according to claim 35, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) is ≧1:1. 50. An intermediate microparticle composition according to claim 49, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) is ≧5:1. 51. An intermediate microparticle composition according to claim 34, wherein the phosphatidylcholine lipid is a C10-20 saturated phosphatidylcholine lipid. 52. An intermediate microparticle composition according to claim 51, wherein the phosphatidylcholine lipid is a 1,2-distearoyl-sn-glycero-3-phosphocholine. 53. An intermediate microparticle composition according to claim 34, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety is a C10-20 saturated phosphatidylethanolamine lipid. 54. An intermediate microparticle composition according to any of claims 34, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety comprises a polyethylene glycol chain with a molecular weight of at least 500. 55. An intermediate microparticle composition according to claim 54, wherein the at least one maleimide moiety is attached to the polyethylene glycol chain. 56. An intermediate microparticle composition according to claim 55, wherein the phosphatidylethanolamine lipid comprising at least one maleimide moiety is a 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000]. 57. An intermediate microparticle composition according to claim 34, wherein the alkoxylated fatty acid is a polyethylene glycol fatty acid ester. 58. An intermediate microparticle composition according to claim 57, wherein the polyethylene glycol fatty acid ester is a C10-20 saturated polyethylene glycol fatty acid ester. 59. An intermediate microparticle composition according to claim 58, wherein the polyethylene glycol fatty acid ester is a PEG40 stearate. 60. An intermediate microparticle composition according to claim 34, wherein the composition is a lyophilisate. 61. An intermediate microparticle composition according to claim 34, wherein the composition is an aqueous dispersion 62. A kit comprising an intermediate composition according to claim 34, and a source of a fluid medium which comprises a physiologically acceptable gas. 63. A kit according to claim 62, wherein the physiologically acceptable gas is selected from air, nitrogen, carbon dioxide, xenon, krypton, sulfur hexafluoride, chlorotrifluoromethane, dichlorodifluoro-methane, bromotrifluoromethane, bromochlorodifluoromethane, tetrafluoromethane, dibromo-difluoromethane, dichlorotetrafluoroethane, chloropentafluoroethane, hexafluoroethane, hexafluoropropylene, octafluoropropane, hexafluoro-butadiene, octafluoro-2-butene, octafluorocyclobutane, decafluorobutane, perfluorocyclopentane, dodecafluoropentane, and tetradecafluorohexane. 64. A kit according to claim 63, wherein the physiologically acceptable gas comprises octafluoropropane. 65. A method of preparing a microparticle composition, the method comprising:
forming a core microparticle structure having a central area and a shell comprising
(i) a phosphatidylcholine lipid;
(ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and
(iii) an alkoxylated fatty acid. 66. A method of preparing a microparticle composition according to claim 65, the method further comprising:
(ii) covalently attaching at least one molecular binding element to the shell of the core microparticle structure. 67. A method of preparing a microparticle composition according to claim 65, wherein step (i) is conducted in the presence of a physiologically acceptable gas. 68. A method of preparing a microparticle composition according to claim 66, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) in step (i) is ≧1:1. 69. A method of preparing a microparticle composition according to claim 68, wherein the conjugation reaction molar ratio of the at least one molecular binding element to component (ii) in step (i) is ≧5:1. 70. A pharmaceutical composition comprising a microparticle composition according to claim 1, and a pharmaceutically acceptable carrier or excipient. 71. A method of molecular imaging comprising:
providing a pharmaceutical composition, the pharmaceutical composition comprising a microparticle composition and a at least one of a pharmaceutically acceptable carrier and excipient, the pharmaceutical composition including a contrast agent, the contrast agent including: i. a phosphatidylcholine lipid; ii. a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and iii. an alkoxylated fatty acid 72. A method of molecular imaging comprising:
providing a microparticle composition, the microparticle composition including a contrast agent, the contrast agent including: iv. a phosphatidylcholine lipid; v. a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and vi. an alkoxylated fatty acid 73. An imaging method comprising:
administering at least one of a microparticle composition and a pharmaceutical composition comprising:
(i) a phosphatidylcholine lipid;
(ii) a phosphatidylethanolamine lipid comprising at least one maleimide moiety; and
(iii) an alkoxylated fatty acid;
and
imaging the subject using at least one imaging technique. 74. An imaging method according to claim 73, wherein the imaging technique comprises ultrasound imaging (US). 75. An imaging method according to claim 73, wherein the imaging technique comprises magnetic resonance imaging (MRI). 76. A microparticle composition suitable for molecular imaging, the composition comprising microparticles, wherein the microparticles comprise: a core microparticle structure having a central area and a shell, and wherein the core microparticle structure comprises:
(i) a phosphatidylcholine lipid; (ii) a phosphatidylethanolamine lipid; and (iii) an alkoxylated fatty acid. | 1,600 |
1,098 | 15,238,357 | 1,612 | Programmable molecular imprinted polymers (MIPs) that have modified binding site kinetics for target imprintable entities (TIEs) that operate to control the adsorption, binding, release and equilibrium distribution of related materials into and out of the MIPs, which are useful for the controlled adsorption, controlled release and control of concentrations of such materials in media including gases, liquids, fluids, biological systems, solutions and other environments. When a collective plurality of the MIPs with modified binding site kinetics are combined, the resulting MIP systems can be tailored to exhibit pseudo zero- and first-order kinetics, as well as higher kinetic profiles, and when further combined with time-delay functionality, can be tailored to exhibit delayed uptake and release, ramped uptake and release of materials, step functions, polynomial, geometric, exponential and other unique kinetic profiles of material exchange between the novel MIPs and a fluid media that are not readily achievable by other means. | 1. A molecularly imprinted polymer system for use in the catch and release of multiple materials comprising:
(a) a first molecularly imprinted polymer with at least one first suboptimal average associative binding constant with respect to a first material to be released; (b) a second molecularly imprinted polymer with at least one second suboptimal average associative binding constant with respect to a second material to be captured; wherein said first molecularly imprinted polymer is dosed with said first material to a desired degree of saturation. wherein said first molecularly imprinted polymer and said second molecularly imprinted polymer are introduced or contacted with a fluid media; and wherein said first and said second molecularly imprinted polymers operate to controllably release a first material into said fluid media and controllably adsorb a second material from said fluid media, respectively. 2. The molecularly imprinted polymer system of claim 1 wherein the said second molecularly imprinted polymer is templated with a second target imprintable entity so as to have an optimal [kTIE] associative binding constant with respect to said second material. 3. The molecularly imprinted polymer system of claim 1 wherein said first suboptimal average associative binding constant differs in magnitude from said optimal [kTIE] associative binding constant by at least one least significant difference (LSD) at an 80% confidence level. 4. The molecularly imprinted polymer system of claim 1 wherein said second suboptimal average associative binding constant differs in magnitude from said optimal [kTIE] associative binding constant by at least one least significant difference (LSD) at an 80% confidence level. 5. The molecularly imprinted polymer system of claim 1 wherein said first and said second suboptimal average associative binding constant differs in magnitude from each other by at least one least significant difference (LSD) at an 80% confidence level. 6. The molecularly imprinted polymer system of claim 1 wherein said first and said second suboptimal average associative binding constant differs in magnitude from each other and also from said optimal [kTIE] associative binding constant by at least one least significant difference (LSD) at an 80% confidence level. 7. A molecularly imprinted polymer system for use in the controlled release of a selected material comprising:
(a) a first molecularly imprinted polymer with at least one first suboptimal average associative binding constant with respect to said selected material; (b) a second molecularly imprinted polymer with at least one second suboptimal average associative binding constant with respect to said selected material; wherein said second suboptimal average associative binding constant differs in magnitude from said first suboptimal average associative binding constant by at least one least significant difference (LSD) at an 80% confidence level; wherein said first molecularly imprinted polymer is dosed with said first material to a desired degree of saturation; wherein said first molecularly imprinted polymer and said second molecularly imprinted polymer are introduced or contacted with a fluid media so as to be in fluidic communication with each other; and wherein said first and said second molecularly imprinted polymers operate to controllably release said selected material into said fluid media following a desired release profile corresponding to the a release rate proportional to the ratio of said first and said second suboptimal average associative binding constants. 8. The molecularly imprinted polymer system of claim 7 further comprising:
(a) a plurality of molecularly imprinted polymers each exhibiting at least one suboptimal average associative binding constant with respect to said selected material;
wherein said suboptimal average associative binding constants of said plurality of molecularly imprinted polymers each exhibit values that differ in magnitude from each other by at least one least significant difference (LSD) at an 80% confidence level;
wherein said plurality of molecularly imprinted polymer is dosed with said selected material to a desired degree of saturation;
wherein said plurality of molecularly imprinted polymers are introduced or contacted with a fluid media so as to be in fluidic communication with each other. 9. The molecularly imprinted polymer system of claim 8 wherein said plurality of molecularly imprinted polymers operate to controllably release said selected material into said fluid media following a desired release profile corresponding to a profile selected from: pseudo-zero order, pseudo-first order, pseudo-n order, exponential, linear, geometric, polynomial, sigmoidal, and combinations thereof. 10. The molecularly imprinted polymer system of claim 7 further comprising a time-delay element associated with at least of one of said plurality of molecularly imprinted polymers; wherein said time delay element operates to delay the time of contact between said associated molecularly imprinted polymer and the fluid media in contact therewith for a selected time period determined by said time delay element; wherein said time-delay element is selected from any suitable material that is slowly or sparingly soluble and/or disintegrates over a desired time period within said fluid media so as to require a desired period of time to be sufficiently dissolved or compromised so as to expose the associated molecularly imprinted polymer to said fluid media. 11. A molecularly imprinted polymer system for use in the treatment of a specific biological pathogen, comprising:
(a) a first molecularly imprinted polymer matrix templated with at least one molecular recognition pattern corresponding to a surface borne molecular entity associated with the exterior cellular membrane of a specific biological pathogen and that operates to bind said pathogen upon contact; (b) a second molecularly imprinted polymer matrix with at least one suboptimum associative binding constant with respect to a treatment agent effective against said biological pathogen; wherein said second molecularly imprinted polymer matrix is preloaded with said treatment agent after formation and extraction of a suitable templating material; (c) optionally, a time-delay coating around said second MIP matrix bearing said preloaded treatment agent; wherein said coating is effective in shielding said second molecularly imprinted polymer matrix for a desired time period; wherein said second molecularly imprinted polymer matrix with said at least one suboptimal associative binding constant operates to controllably release the preloaded treatment agent at a controlled rate into a fluid media. 12. The molecularly imprinted polymer system of claim 11, further comprising a third molecularly imprinted polymer matrix; wherein said third molecularly imprinted polymer matrix has been templated with the treatment agent to exhibit a higher associative binding constant than that of said second molecularly imprinted polymer matrix and operates to adsorb excess treatment agent from said surrounding fluid media. 13. The molecularly imprinted polymer system of claim 11, further comprising a second delay-release coating around said third molecularly imprinted polymer matrix; wherein said coating is effective in shielding said third molecularly imprinted polymer matrix for a desired second time period that is greater than or equal to the time period exhibited by said delay-release coating around said second molecularly imprinted polymer matrix. 14. The molecularly imprinted polymer system of claim 11 wherein said first, second and third molecularly imprinted polymer matrix components are combined into a molecularly imprinted polymer system by means of one or more tethering elements; wherein said tethering element is selected from a physical link, a chemical bond, a molecular bond, a molecular linker group, a polymer chain, an ionic bond, a physical linker moiety, and combinations thereof. 15. The molecularly imprinted polymer system of claim 14, wherein said physical linker moiety comprises a molecule having at least two or more template groups (T) and at least one spacer group (S); wherein said template group is any molecule or molecular fragment capable of being used as a target imprinted entity (TIE) in the formation of a molecularly imprinted polymer matrix; and wherein said spacer group is any molecule or molecular fragment that can be formed into a linear chain or repeating chemical unit; wherein said physical linker moiety has the following structure:
T−(S)n −T
wherein n includes any integer value from n=1 to about 1000 and wherein said template groups operate to bind to a molecularly imprinted polymer that has been imprinted with a target imprinted entity comprising a template group, a chemically modified template group, a molecular analog to said template group bearing at least one common molecular recognition site, and combinations thereof. | Programmable molecular imprinted polymers (MIPs) that have modified binding site kinetics for target imprintable entities (TIEs) that operate to control the adsorption, binding, release and equilibrium distribution of related materials into and out of the MIPs, which are useful for the controlled adsorption, controlled release and control of concentrations of such materials in media including gases, liquids, fluids, biological systems, solutions and other environments. When a collective plurality of the MIPs with modified binding site kinetics are combined, the resulting MIP systems can be tailored to exhibit pseudo zero- and first-order kinetics, as well as higher kinetic profiles, and when further combined with time-delay functionality, can be tailored to exhibit delayed uptake and release, ramped uptake and release of materials, step functions, polynomial, geometric, exponential and other unique kinetic profiles of material exchange between the novel MIPs and a fluid media that are not readily achievable by other means.1. A molecularly imprinted polymer system for use in the catch and release of multiple materials comprising:
(a) a first molecularly imprinted polymer with at least one first suboptimal average associative binding constant with respect to a first material to be released; (b) a second molecularly imprinted polymer with at least one second suboptimal average associative binding constant with respect to a second material to be captured; wherein said first molecularly imprinted polymer is dosed with said first material to a desired degree of saturation. wherein said first molecularly imprinted polymer and said second molecularly imprinted polymer are introduced or contacted with a fluid media; and wherein said first and said second molecularly imprinted polymers operate to controllably release a first material into said fluid media and controllably adsorb a second material from said fluid media, respectively. 2. The molecularly imprinted polymer system of claim 1 wherein the said second molecularly imprinted polymer is templated with a second target imprintable entity so as to have an optimal [kTIE] associative binding constant with respect to said second material. 3. The molecularly imprinted polymer system of claim 1 wherein said first suboptimal average associative binding constant differs in magnitude from said optimal [kTIE] associative binding constant by at least one least significant difference (LSD) at an 80% confidence level. 4. The molecularly imprinted polymer system of claim 1 wherein said second suboptimal average associative binding constant differs in magnitude from said optimal [kTIE] associative binding constant by at least one least significant difference (LSD) at an 80% confidence level. 5. The molecularly imprinted polymer system of claim 1 wherein said first and said second suboptimal average associative binding constant differs in magnitude from each other by at least one least significant difference (LSD) at an 80% confidence level. 6. The molecularly imprinted polymer system of claim 1 wherein said first and said second suboptimal average associative binding constant differs in magnitude from each other and also from said optimal [kTIE] associative binding constant by at least one least significant difference (LSD) at an 80% confidence level. 7. A molecularly imprinted polymer system for use in the controlled release of a selected material comprising:
(a) a first molecularly imprinted polymer with at least one first suboptimal average associative binding constant with respect to said selected material; (b) a second molecularly imprinted polymer with at least one second suboptimal average associative binding constant with respect to said selected material; wherein said second suboptimal average associative binding constant differs in magnitude from said first suboptimal average associative binding constant by at least one least significant difference (LSD) at an 80% confidence level; wherein said first molecularly imprinted polymer is dosed with said first material to a desired degree of saturation; wherein said first molecularly imprinted polymer and said second molecularly imprinted polymer are introduced or contacted with a fluid media so as to be in fluidic communication with each other; and wherein said first and said second molecularly imprinted polymers operate to controllably release said selected material into said fluid media following a desired release profile corresponding to the a release rate proportional to the ratio of said first and said second suboptimal average associative binding constants. 8. The molecularly imprinted polymer system of claim 7 further comprising:
(a) a plurality of molecularly imprinted polymers each exhibiting at least one suboptimal average associative binding constant with respect to said selected material;
wherein said suboptimal average associative binding constants of said plurality of molecularly imprinted polymers each exhibit values that differ in magnitude from each other by at least one least significant difference (LSD) at an 80% confidence level;
wherein said plurality of molecularly imprinted polymer is dosed with said selected material to a desired degree of saturation;
wherein said plurality of molecularly imprinted polymers are introduced or contacted with a fluid media so as to be in fluidic communication with each other. 9. The molecularly imprinted polymer system of claim 8 wherein said plurality of molecularly imprinted polymers operate to controllably release said selected material into said fluid media following a desired release profile corresponding to a profile selected from: pseudo-zero order, pseudo-first order, pseudo-n order, exponential, linear, geometric, polynomial, sigmoidal, and combinations thereof. 10. The molecularly imprinted polymer system of claim 7 further comprising a time-delay element associated with at least of one of said plurality of molecularly imprinted polymers; wherein said time delay element operates to delay the time of contact between said associated molecularly imprinted polymer and the fluid media in contact therewith for a selected time period determined by said time delay element; wherein said time-delay element is selected from any suitable material that is slowly or sparingly soluble and/or disintegrates over a desired time period within said fluid media so as to require a desired period of time to be sufficiently dissolved or compromised so as to expose the associated molecularly imprinted polymer to said fluid media. 11. A molecularly imprinted polymer system for use in the treatment of a specific biological pathogen, comprising:
(a) a first molecularly imprinted polymer matrix templated with at least one molecular recognition pattern corresponding to a surface borne molecular entity associated with the exterior cellular membrane of a specific biological pathogen and that operates to bind said pathogen upon contact; (b) a second molecularly imprinted polymer matrix with at least one suboptimum associative binding constant with respect to a treatment agent effective against said biological pathogen; wherein said second molecularly imprinted polymer matrix is preloaded with said treatment agent after formation and extraction of a suitable templating material; (c) optionally, a time-delay coating around said second MIP matrix bearing said preloaded treatment agent; wherein said coating is effective in shielding said second molecularly imprinted polymer matrix for a desired time period; wherein said second molecularly imprinted polymer matrix with said at least one suboptimal associative binding constant operates to controllably release the preloaded treatment agent at a controlled rate into a fluid media. 12. The molecularly imprinted polymer system of claim 11, further comprising a third molecularly imprinted polymer matrix; wherein said third molecularly imprinted polymer matrix has been templated with the treatment agent to exhibit a higher associative binding constant than that of said second molecularly imprinted polymer matrix and operates to adsorb excess treatment agent from said surrounding fluid media. 13. The molecularly imprinted polymer system of claim 11, further comprising a second delay-release coating around said third molecularly imprinted polymer matrix; wherein said coating is effective in shielding said third molecularly imprinted polymer matrix for a desired second time period that is greater than or equal to the time period exhibited by said delay-release coating around said second molecularly imprinted polymer matrix. 14. The molecularly imprinted polymer system of claim 11 wherein said first, second and third molecularly imprinted polymer matrix components are combined into a molecularly imprinted polymer system by means of one or more tethering elements; wherein said tethering element is selected from a physical link, a chemical bond, a molecular bond, a molecular linker group, a polymer chain, an ionic bond, a physical linker moiety, and combinations thereof. 15. The molecularly imprinted polymer system of claim 14, wherein said physical linker moiety comprises a molecule having at least two or more template groups (T) and at least one spacer group (S); wherein said template group is any molecule or molecular fragment capable of being used as a target imprinted entity (TIE) in the formation of a molecularly imprinted polymer matrix; and wherein said spacer group is any molecule or molecular fragment that can be formed into a linear chain or repeating chemical unit; wherein said physical linker moiety has the following structure:
T−(S)n −T
wherein n includes any integer value from n=1 to about 1000 and wherein said template groups operate to bind to a molecularly imprinted polymer that has been imprinted with a target imprinted entity comprising a template group, a chemically modified template group, a molecular analog to said template group bearing at least one common molecular recognition site, and combinations thereof. | 1,600 |
1,099 | 13,386,349 | 1,643 | A delivery vehicle, for delivering a pharmaceutically active agent or a marker to a cell, comprising a ligand binding portion specific for a Fas Ligand, and a carrier for the pharmaceutically active agent or marker. | 1. A delivery vehicle comprising a ligand binding portion specific for a Fas Ligand, and a carrier for a pharmaceutically active agent or marker. 2. A delivery vehicle as claimed in claim 1, wherein the carrier is a microparticle, nanoparticle, microcapsule, microsphere, micelle or liposome. 3-4. (canceled) 5. A delivery vehicle as claimed in claim 1, wherein the ligand binding portion specific for a Fas Ligand comprises or consists of:
(a) a Fas receptor or a derivative thereof; (b) a full-length Fas protein, or a fragment thereof; (c) an extracellular domain of a Fas protein, or a fragment thereof; (d) a ligand binding domain of a Fas protein; (e) a peptide, a protein, an aptamer, an antibody, an antibody fragment, a fusion protein or a chimeric protein; or (f) a Fas protein, or a fragment thereof, fused to a fragment crystallizable region (Fc region) of an immunoglobulin to form a chimeric fusion protein. 6-7. (canceled) 8. A delivery vehicle as claimed in claim 1, wherein the ligand binding portion is human or murine. 9-10. (canceled) 11. A delivery vehicle as claimed in claim 1, wherein the ligand binding portion is coupled to the carrier via a linking molecule. 12. A delivery vehicle as claimed in claim 11, wherein the linking molecule is a fragment crystallizable region (Fc) of an immunoglobulin. 13. A delivery vehicle as claimed in claim 1, wherein the pharmaceutically active agent, marker or drug is preferably not an agent capable of specific binding to a Fas Ligand. 14. (canceled) 15. A delivery vehicle as claimed in claim 1, wherein the delivery vehicle is a microparticle containing the pharmaceutically active agent or marker, and wherein the microparticle is covalently or non-covalently associated with Fas, a derivative thereof, or a specific binding molecule for Fas Ligand. 16-25. (canceled) 26. A method of treating a disease or medical condition, comprising administering to a subject a delivery vehicle, wherein the delivery vehicle is a carrier for a pharmaceutically active agent bound to a ligand binding portion specific for Fas ligand, and wherein the carrier includes an effective amount of the pharmaceutically active agent. 27. A method as claimed in claim 26, wherein the disease or condition is a brain tumour, ovarian cancer, prostate cancer, breast cancer, an intraperitoneal tumour, an ovarian tumour, a gastrointestinal tumour, colon cancer, lung cancer, pancreatic cancer or a cancer type or tumour where the Fas Ligand is expressed in the tumour cells. 28. A method as claimed in claim 26, wherein the disease or condition is a neurological disease, optionally, motor neuron disease, Alzheimer's disease, Parkinson's disease, a neuropathic pain syndrome or a peripheral nerve or spinal cord injury. 29. (canceled) 30. A method of diagnosing a disease comprising detecting Fas Ligand-expressing cells by administering to a subject a delivery vehicle, wherein the delivery vehicle is a carrier for a marker bound to a ligand binding portion specific for Fas ligand, and wherein the marker is attached to, or contained or encapsulated by the carrier. 31. The method as claimed in claim 30, wherein the method of diagnosing a disease, is a method of diagnosing a neoplasm or neurological disorder. 32. A pharmaceutical composition comprising a delivery vehicle as claimed in claim 1 and one or more of a physiologically or pharmaceutically acceptable carrier, excipient, or stabilizer. 33. A method for preparing a delivery vehicle comprising forming a microparticle containing or encapsulating a pharmaceutically active agent, marker or drug and attaching a ligand binding portion specific for Fas Ligand to said microparticle. 34. A microparticle containing a pharmaceutically active substance, wherein the microparticle is covalently or non-covalently associated with Fas, a derivative thereof, or a specific binding molecule for Fas Ligand. 35-44. (canceled) 45. A method for the delivery into a cell of a pharmaceutically active agent or marker comprising administering a delivery vehicle to the cell, wherein the delivery vehicle is a carrier bound to a ligand binding portion specific for a Fas ligand, and wherein the agent or marker is covalently or noncovalently associated with the carrier. 46-53. (canceled) 54. A drug conjugate comprising a drug and: (i) Fas, (ii) a derivative of Fas, or (iii) a specific binding molecule for Fas Ligand. 55. A drug conjugate according to claim 54, wherein the Fas derivative is full-length Fas, FasFc or a fusion protein of Fas. 56. A drug conjugate according to claim 54, wherein the specific binding molecule for Fas Ligand is a peptide, a protein, an aptamer, an antibody or an antibody fragment. 57. (canceled) | A delivery vehicle, for delivering a pharmaceutically active agent or a marker to a cell, comprising a ligand binding portion specific for a Fas Ligand, and a carrier for the pharmaceutically active agent or marker.1. A delivery vehicle comprising a ligand binding portion specific for a Fas Ligand, and a carrier for a pharmaceutically active agent or marker. 2. A delivery vehicle as claimed in claim 1, wherein the carrier is a microparticle, nanoparticle, microcapsule, microsphere, micelle or liposome. 3-4. (canceled) 5. A delivery vehicle as claimed in claim 1, wherein the ligand binding portion specific for a Fas Ligand comprises or consists of:
(a) a Fas receptor or a derivative thereof; (b) a full-length Fas protein, or a fragment thereof; (c) an extracellular domain of a Fas protein, or a fragment thereof; (d) a ligand binding domain of a Fas protein; (e) a peptide, a protein, an aptamer, an antibody, an antibody fragment, a fusion protein or a chimeric protein; or (f) a Fas protein, or a fragment thereof, fused to a fragment crystallizable region (Fc region) of an immunoglobulin to form a chimeric fusion protein. 6-7. (canceled) 8. A delivery vehicle as claimed in claim 1, wherein the ligand binding portion is human or murine. 9-10. (canceled) 11. A delivery vehicle as claimed in claim 1, wherein the ligand binding portion is coupled to the carrier via a linking molecule. 12. A delivery vehicle as claimed in claim 11, wherein the linking molecule is a fragment crystallizable region (Fc) of an immunoglobulin. 13. A delivery vehicle as claimed in claim 1, wherein the pharmaceutically active agent, marker or drug is preferably not an agent capable of specific binding to a Fas Ligand. 14. (canceled) 15. A delivery vehicle as claimed in claim 1, wherein the delivery vehicle is a microparticle containing the pharmaceutically active agent or marker, and wherein the microparticle is covalently or non-covalently associated with Fas, a derivative thereof, or a specific binding molecule for Fas Ligand. 16-25. (canceled) 26. A method of treating a disease or medical condition, comprising administering to a subject a delivery vehicle, wherein the delivery vehicle is a carrier for a pharmaceutically active agent bound to a ligand binding portion specific for Fas ligand, and wherein the carrier includes an effective amount of the pharmaceutically active agent. 27. A method as claimed in claim 26, wherein the disease or condition is a brain tumour, ovarian cancer, prostate cancer, breast cancer, an intraperitoneal tumour, an ovarian tumour, a gastrointestinal tumour, colon cancer, lung cancer, pancreatic cancer or a cancer type or tumour where the Fas Ligand is expressed in the tumour cells. 28. A method as claimed in claim 26, wherein the disease or condition is a neurological disease, optionally, motor neuron disease, Alzheimer's disease, Parkinson's disease, a neuropathic pain syndrome or a peripheral nerve or spinal cord injury. 29. (canceled) 30. A method of diagnosing a disease comprising detecting Fas Ligand-expressing cells by administering to a subject a delivery vehicle, wherein the delivery vehicle is a carrier for a marker bound to a ligand binding portion specific for Fas ligand, and wherein the marker is attached to, or contained or encapsulated by the carrier. 31. The method as claimed in claim 30, wherein the method of diagnosing a disease, is a method of diagnosing a neoplasm or neurological disorder. 32. A pharmaceutical composition comprising a delivery vehicle as claimed in claim 1 and one or more of a physiologically or pharmaceutically acceptable carrier, excipient, or stabilizer. 33. A method for preparing a delivery vehicle comprising forming a microparticle containing or encapsulating a pharmaceutically active agent, marker or drug and attaching a ligand binding portion specific for Fas Ligand to said microparticle. 34. A microparticle containing a pharmaceutically active substance, wherein the microparticle is covalently or non-covalently associated with Fas, a derivative thereof, or a specific binding molecule for Fas Ligand. 35-44. (canceled) 45. A method for the delivery into a cell of a pharmaceutically active agent or marker comprising administering a delivery vehicle to the cell, wherein the delivery vehicle is a carrier bound to a ligand binding portion specific for a Fas ligand, and wherein the agent or marker is covalently or noncovalently associated with the carrier. 46-53. (canceled) 54. A drug conjugate comprising a drug and: (i) Fas, (ii) a derivative of Fas, or (iii) a specific binding molecule for Fas Ligand. 55. A drug conjugate according to claim 54, wherein the Fas derivative is full-length Fas, FasFc or a fusion protein of Fas. 56. A drug conjugate according to claim 54, wherein the specific binding molecule for Fas Ligand is a peptide, a protein, an aptamer, an antibody or an antibody fragment. 57. (canceled) | 1,600 |
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