The invention concerns delivery particles (DP""s) for active principles (AP""s) based on linear amphiphilic polyamino-acids (PAA""s), with xcex1-peptide chains, capable of being spontaneously formed by contacting PAA""s with a liquid medium, preferably with water, wherein the hydrophile part of the PAA""s is solubilized more than the hydrophobic parts of said PAA""s, such that the latter precipitate while being organised in discrete supra-molecular arrangements, of average size ranging between 0.01 and 20 xcexcm, capable of combining with at least an AP and releasing the latter in vivo, in prolonged and controlled manner. The inventive suspension is characterised in that the recurrent amino acids (rAA""s) constituting the main chain of PAA""s are identical to or different from one another and are glutamic acid and/or aspartic acid and/or their salts; and some of said rAA""s bear at least one hydrophobic group, said hydrophobic group, being identical to or different from one another. The invention is useful as carriers for active principles, in particular pharmaceutical, insulin or for therapeutic uses.

This application is a 371 of PCT/FR99/02859, filed on Nov. 19, 1999.
The field of the present invention is that of Delivery Particles (DPs) which can be used for the administration of active principles (APs). The latter are preferably medicinal products or nutrients for adminstration to an animal or human organism via the oral, nasal, vaginal, ocular, subcutaneous, intravenous, intramuscular, intradermal, intraperitoneal, intracerebral, parenteral, etc. route. However, they may also be plant-protection products, such as herbicides, pesticides, insecticides, fungicides, etc. In terms of chemical nature, the APs with which the invention is more particularly, but not exclusively, concerned are, for example, proteins, glycoproteins, peptides, polysaccharides, lipopolysaccharides, oligonucleotides and polynucleotides.
The present invention relates more specifically to Delivery Particles, advantageously of the submicron type, based on polyamino acids (PAAs). The present invention is directed toward both naked particles per se, and AP vector systems consisting of particles loaded with the AP(s) under consideration. The present invention also relates to aqueous colloidal suspensions comprising these DPs.
The invention also relates to processes for preparing said particles and colloidal suspensions, with and without APs.
The aim of the encapsulation of APs in DPs is in particular to increase the bioavailability of said APs. Many encapsulation techniques have already been proposed. Such techniques are directed, firstly, toward enabling the AP to be transported to its site of therapeutic action, while at the same time protecting it against the body""s attacks (hydrolysis, enzymatic digestion, etc.) and, secondly, toward controlling the release of the AP over its site of action, in order to maintain the amount available to the organism at the desired level. The APs with which these misadventures of delivery and residence in the body are concerned are, for example, proteins, but may also be any other products, of synthetic or natural origin. The review by M. J. HUMPHREY (Delivery System for peptide Drugs, edited by S. DAVIS and L. ILLUM, Plenum Press, N.Y., 1986), gives an account of the problem concerning the improvement of the bioavailability of APs and the advantage of systems for delivery and controlled release.
Of all the materials which can be envisaged for making up DPs, polymers are increasingly used on account of their intrinsic properties. As regards the list of specifications which it is desired to obtain for such DPs, this is particularly demanding and comprises, in particular, the following specifications.
1xe2x80x94It should advantageously be possible to obtain a particle size distribution which is controlled and suitable for the mode of administration chosen and/or for the therapeutic site targeted.
2xe2x80x94It is desirable for the DPs to protect the AP until the site of release is reached.
3xe2x80x94The DPs should advantageously control the rate of release of the AP.
4xe2x80x94It is preferable for the polymer which constitutes the DPs to be biocompatible, able to be eliminated (by excretion) and/or biodegradable and, better still, for it to be metabolized into products which are nontoxic to the body.
5xe2x80x94It is also advantageous for the polymer which constitutes the DPs not to induce an immune response.
6xe2x80x94Finally, it is also desirable for it to be possible to obtain the DPs and the DP-AP systems using a process which does not denature the AP.
The prior technical propositions, described above, have attempted to satisfy this set of specifications. By way of illustration, mention will be made of prior propositions (a) to (h): according to a first approach, which comprises propositions (a) to (d), the inclusion of the active principle takes place during the formation of the delivery supports; according to a second approach, mentioned in propositions (e) to (h), DPs are produced which are capable, once manufactured, of associating spontaneously by absorption to the AP.
(a) Patent U.S. Pat. No. 5,286,495 relates to a process of encapsulation by spraying proteins in aqueous phase, using materials having opposite charges, namely: alginate (negatively charged) and polylysine (positively charged). This manufacturing process makes it possible to produce particles greater than 35 xcexcm in size.
(b) In addition, emulsion techniques are commonly used for preparing microparticles loaded with AP. For example, patent applications WO 91/06286, WO 91/06287 and WO 89/08449 disclose such emulsion techniques in which use is made of organic solvents in order to solubilize polymers, for example of the polylactic type. However, the solvents have proven to be possibly denaturing, in particular for peptide or polypeptide APs.
(c) Biocompatible DPs formed in aqueous solution and called proteinoids, described as early as 1970 by W. FOX and K. DOSE in xe2x80x9cMolecular Evolution and the origin of Lifexe2x80x9d, Ed. Marcel DEKKER Inc. (1977), are also known. Thus, patent application WO 88/01213 provides a system based on a mixture of artificial polypeptides obtained by thermal condensation of amino acids. The microparticles according to that invention are obtained by changing the pH, which causes precipitation of the proteinoid particles.
(d) Mention will also be made, as a matter of interest, of U.S. Pat. Nos. 4,351,337 and 4,450,150. which are the product of a field other than that of the delivery of APs, characteristic of the invention. These patents disclose implants of masses attached and located at quite precise sites in the body. The implants are produced from polymeric materials of the poly-xcex1-amino acid type (Leu/GluOH and GluOEt/GluOH, respectively). According to the teaching of that patent, the preferred polyamino acids are those which are rich in hydrophobic amino acid (eg. more than 50% of Leucine or of Glu OEt) and water-insoluble. The AP can be incorporated into a solution of poly-xcex1-amino acid in an organic solvent. This solution is that used to form the implant by molding/drying (evaporation). According to another variant, the AP can be included in the core of a microcapsule, the case of which is obtained using a copolymer solution, and the diameter of which is greater than or equal to 5000 xcexcm. The core can consist of pure AP or of a copolymer matrix which includes the AP, and can be obtained from the solution of PAA in an organic solvent.
(e) PCT patent application WO/FR97/02810 discloses a composition for the controlled release of active principles, comprising a plurality of lamellar particles of a biodegradable polymer, at least partly crystalline (lactic acid polymer), and of an active principle absorbed onto said particles. The release of the active principle takes place by desorption.
(f) The publication xe2x80x9cCHEMISTRY LETTERS 1995, 707, AKIYOSHI et alxe2x80x9d relates to the stabilization of insulin by supramolecular complexation with nanoparticles formed from ten or so polysaccharide chains made hydrophobic by grafting cholesterol.
(g) The article published in xe2x80x9cMACROMOLECULES 1997, 30, 4013-4017xe2x80x9d describes copolymers composed of a peptide block based on L-phenylalanine, on xcex3-benzyl-L-glutamate or on O-(tetra-O-acetyl-xcex2-D-glucopyranosyl)-L-serine, and a synthetic block, such as poly(2-methyl-2-oxazoline) or poly(2-phenyl-2-oxazoline). Some of these polymers aggregate in aqueous medium to form particles of 400 nm, capable of associating with an enzyme, lipase.
(h) The subject of patent FR 95-03978 is polyamino acid particles which can be used for delivering active principles and which are characterized in that their constituent polyamino acids comprise at least two types of recurrent amino acids, AANs (which are neutral and hydrophobic) and AAIs (which are ionizable and hydrophilic). The particles are obtained spontaneously by dispersion of the polyamino acid powder in an aqueous solution. The particles thus obtained associate spontaneously in aqueous suspension with active principles, which are for example protein in nature.
These prior technical propositions more or less satisfy the specifications of the list of specifications indicated above. However, such delivery particles for active principles remain perfectible.
Given this irrefutable fact, one of the essential objectives of the present invention is to improve the prior technical proposition referenced (h) FR 95-03 978, by providing novel DPs based on linear, amphiphilic poly-xcex1-amino acids (PAAs), the hydrophilic and hydrophobic natures of which are, respectively, provided by the amino acids of the main chain of the polymer and by hydrophobic radicals attached laterally to a fraction of the amino acids via a covalent attachment. This improvement concerns more precisely a means of controlling the characteristics of the association between the APs and DPs in order to improve the degree of loading of AP, and/or to improve the kinetics of release of the AP.
Another essential objective of the invention is to provide an aqueous colloidal suspension of delivery particles for active principles, which comprises PAA particles which satisfy the specifications targeted above and which constitutes a pharmaceutical form which is appropriate and suitable for administration, for example oral administration, to humans or animals.
Another essential objective of the invention is to provide a process for preparing PAA particles which can be used in particular as vectors of active principles, said process having to be even more economical, simpler to carry out and nondenaturing for the active principles, and also still having to allow fine control of the mean particle size of the particles obtained.
Another essential objective of the invention is the use of the abovementioned suspensions and particles for preparing medicinal products (e.g. vaccines) and/or nutrients, in particular for administration, especially oral, nasal, vaginal, ocular, subcutaneous, intravenous, intramuscular, intradermal, intraperitoneal, intracerebral or parenteral administration, of active principles such as proteins, glycoproteins, peptides, polysaccharides, lipopolysaccharides, oligonucleotides or polynucleotides.
Another essential objective of the present invention is to provide suspensions of DPs which are partly submicron and micron particles, which are based on PAA and which can be used as vectors for an AP, in particular medicinal and/or nutrient APs, for the administration of said AP to a human or animal organism.
Another objective of the present invention is to provide a medicinal product, such as a system for sustained release of active principles, which is easy and economical to produce and which is also biocompatible and capable of ensuring a very high level of bioavailability of the AP.
Another essential objective of the invention is to provide a system for delivering vaccine, which is nonimmunogenic intrinsically and in combination with one or more antigens.
The objectives relating to the products, inter alia, are achieved through the present invention which relates, first of all, to particles which can be used in particular for the delivery (DPs) of active principle(s), which are of the type of those:
based on amphiphilic, linear polyamino acids (PAAs) with xcex1-peptide chains;
capable of forming spontaneously by bringing the PAAs into contact with a liquid medium, preferably with water, in which the hydrophilic portion of the PAAs solubilizes more than the hydrophobic portion of said PAAs, such that the latter precipitate while becoming organized in discrete supramolecular arrangements;
having a mean size of less than 200 xcexcm, preferably less than 100 xcexcm, and even more preferably of between 0.01 and 20 xcexcm;
and capable of associating with at least one AP and of releasing the latter in vivo, in a sustained and controlled way;
xe2x80x83characterized:
in that the recurrent amino acids (rAAs) constituting the main chain of the PAAs are identical to or different from one another, and are chosen from amino acids of acid type, and preferably from the group comprising glutamic acid and/or aspartic acid and/or the salts thereof, respectively glutamates and aspartates,
and in that some of these rAAs each bear at least one hydrophobic R0 group, these hydrophobic R0 groups being identical to or different from one another.
One of the conditions for the particles according to the invention to form is that the hydrophilic portion of the PAAs solubilizes more in the liquid suspension medium than the hydrophobic portion thereof.
The recurrent amino acids (rAAs) selected in accordance with the invention are amino acids containing carboxylic functions (Glu and Asp) in COOH form or in the form of salts (carboxylates) COOxe2x88x92, X+; with X corresponding to alkali metals, preferably to Na.
It is to the applicant""s credit to have chosen, by way of material constituting the DPs, a particular class of poly-xcex1-amino acids, the (co)monomers of which are recurrent amino acids (rAAs) of polar nature, and which have been made amphiphilic by modifying the hydrophilic nature by adding hydrophobic side chains to a fraction of the rAAs. This acquired amphiphilic nature confers on the PAAs the possibility of forming colloidal suspensions of DPs, compatible with the pH of the physiological media encountered in the applications intended.
This selection advantageously makes it possible to have a greater choice regarding the nature of the hydrophobic groups, and thus a better means of controlling the hydrophobicity of the polymer and of the DP, thereby making it possible to optimize the association and the release of the APs.
The structure of the PAA polymer, the nature of the amino acids Asp and Glu and the hydrophobic radicals are chosen such that the polymer chains spontaneously become structured in the form of particles which are small in size and stable in physiological medium.
The structure of the PAA polymer, the nature of the amino acids Asp and Glu and the hydrophobic radicals are chosen such that the DPs encapsulate proteins, or other APs, in aqueous medium via a mechanism which is spontaneous and nondenaturing for proteins.
The structure of the PAA polymer, the nature of the amino acids Asp and Glu and the hydrophobic radicals are chosen such that the DPs release the APs into physiological medium and, more precisely, in vivo, the kinetics of release depending on the nature of the polymer and of the DPs which it is capable of forming.
Without it being limiting, this selection is directed, more specifically, toward PAAs, the main chain of which consists of rAAs which are identical to one another. The primary structure of the PAAs can be of the ordered, alternating block type (block PAAs) or of the disordered, random block type (random PAAs) According to a preferred characteristic of the invention, the PAAs are polymers containing up to approximately 1000 rAAs, and preferably up to approximately 500 rAAs, and even more preferably up to approximately 200 rAAs.
The hydrophobic R0 groups participate in the aggregation of the polymeric chains, which is at the heart of the formation of the DPs. According to a preferred arrangement of the invention, these R0 groups are identical to or different from one another, and are selected from the group comprising:
(i) linear or branched, preferably linear, C-C20, and even more preferably C2-C18, alkyls, acyls or alkenyls;
(ii) hydrocarbon-based groups containing one or more heteroatoms, preferably those containing oxygen and/or sulfur, and even more preferably those of the following formula:
xe2x80x83in which:
the R1 radicals are identical to or different from one another, and correspond to hydrogen or to a radical satisfying the same definition as that given above in point (i),
q=1 to 100;
(iii) aryls, aralkyls or alkylaryls, preferably aryls;
(iv) natural hydrophobic derivatives, preferably cholesterol, phosphatidylcholines and diacylglycerols.
For the purpose of the present invention, the term xe2x80x9chydrocarbon-based groupsxe2x80x9d is intended to mean groups comprising in particular hydrogen and carbon atoms.
Preferably, the R0 groups are selected from the following group of radicals: methyl, ethyl, propyl, dodecyl, hexadecyl, octadecyl.
In accordance with the invention, each hydrophobic R0 group is attached to the main chain via a covalent attachment which can be cleaved, preferably by chemical and/or enzymatic hydrolysis, said attachment being more especially of the ester or amide type.
Advantageously, the fraction of rAA monomers bearing hydrophobic groups represents a proportion greater than or equal to 3%, preferably between 3-70%, and even more preferably between 13 and 60%.
In practice, the PAAs constituting the DPs of the suspension according to the invention are, for example:
polymers based on glutamate:
poly(sodium glutamate)-block-(methyl glutamate),
poly(sodium glutamate)-block-(ethyl glutamate),
poly(sodium glutamate)-block-(propyl glutamate),
poly(sodium glutamate)-block-(octadecyl glutamate),
poly(sodium glutamate)-block-(benzyl glutamate),
poly(sodium glutamate)-co-(methyl glutamate),
poly(sodium glutamate)-co-(ethyl glutamate),
poly(sodium glutamate)-co-(propyl glutamate),
poly(sodium glutamate)-co-(dodecyl glutamate),
poly(sodium glutamate)-block-(octadecyl glutamate),
polymers based on aspartate:
poly(sodium aspartate)-block-(methyl aspartate),
poly(sodium aspartate)-block-(ethyl aspartate),
poly(sodium aspartate)-block-(propyl aspartate),
poly(sodium aspartate)-block-(octadecyl aspartate),
poly(sodium aspartate)-block-(benzyl aspartate).
Advantageously, the DP particles have a mean size of between 0.01 xcexcm and 0.5 xcexcm, preferably between 0.01 and 0.2 xcexcm. For the purpose of the invention, the term xe2x80x9cmean sizexe2x80x9d or xe2x80x9cmean particle sizexe2x80x9d is intended to mean the arithmetic mean of the diameters by volume [D4,3], established by laser scattering, and the diameter of gyration measured by elastic light scattering.
One of the assets of the invention is that it has achieved very good control of the mean particle size of these matricial entities and of their particle size distribution. This control involves attaining particle sizes which are extremely small, of the order of a few nanometers, and of very low polydispersity, in the knowledge that it is possible to increase the size of these nanoparticles by aggregation.
The control of the DP size takes place via the composition of rAA and R0 of the polyamino acids, but also, for the same composition, through the choice of a block structure and of the production process.
According to an alternative regarding the size of the DPs of the suspension according to the invention, said delivery particles DP are aggregated in order to obtain new particles which are bigger in size.
The PAA material constituting the DPs according to the invention is preferably a homopolymer or a copolymer in which the rAA=Asp and/or Glu, and which has been made amphiphilic by modifying the hydrophilic nature of this rAA by adding hydrophobic R0 side chains to a fraction of the rAAs. This acquired amphiphilic to a fraction of the rAAs. This acquired amphiphilic nature makes it possible to bring together at least three novel and surprising properties:
1. the first is the possibility of spontaneously forming colloidal suspensions of DPs compatible with the pH of the physiological media encountered in the therapeutic applications intended;
2. the second is the spontaneous association of the DPs with Ads in the absence of an agent other than water, water being used as solvent of the DPs, and water being not denaturing for the APs in the case where APs are proteins;
3. the third is the possibility of releasing the AP of the AP-DP association complex, under physiological conditions, with pharmacokinetic and pharmacodynamic profiles which allow uses in the therapeutic domain to be envisaged.
The APs, in particular the APs which are protein in nature, are in fact trapped by the delivery particles DP, which can be compared to matrices made of water and of PAAs in which the AP(s) are dispersed. Their release takes place either by spontaneous dissociation, or in the course of the degradation of the PAAs and of the subsequent destructuring of the particles. The trapping of the Ads is produced spontaneously by simply mixing them in an aqueous colloidal suspension of DPs according to the invention.
The particles DP loaded with at least one active principle AP constitute another subject of the present invention.
According to another of its aspects, upstream of the particles, which form its essential subject, the invention also relates to the precursors of the DPs, which precursors are the PAAs selected, as defined above.
Downstream of the DPs, the invention also encompasses the colloidal, preferably aqueous, suspension of particles as defined above. controlled release, of said DP in vivo, for therapeutic purposes.
The invention relates, moreover, to a process for preparing the abovementioned colloidal suspension of particles, e.g. of delivery particles (DPs) for active principles (APs),
characterized in that it consists, essentially:
Ixe2x80x94in using linear PAAs containing xcex1-peptide chains of recurrent amino acids (rAAs) selected from natural polar amino acids, namely glutamic acid and aspartic acid, and/or from the salts thereof, respectively glutamates and aspartates; some of these rAAs each bearing at least one hydrophobic R0 group, these R0 groups being identical to or different from one another;
IIxe2x80x94in placing the amphiphilic PAAs in a solvent, preferably aqueous, medium;
III and then in making at least the rAAs carrying R0 insoluble in the medium, so as to precipitate these rAAs and thus to form discrete supramolecular arrangements.
According to an advantageous variant of this process, at least one active principle is incorporated into the liquid medium containing the particles DP, so as to obtain a colloidal suspension of DPs loaded with active principle(s).
In addition, in order to increase the size of the particles, it is conceivable to anticipate, in this process for preparing the suspension, at least one additional step of aggregation of the particles, using at least one aggregation agent consisting of a salt and/or an acid and/or a base and/or a polymer, optionally an ionic polymer (eg. polylysine, polyethyleneimine, etc.). For greater detail, reference will be made to patent application FR 95-03 918.
After having gone through the essential characteristics of the invention in terms of particles, of PAA precursors of said particles, of a colloidal suspension of these particles and of preparation of this suspension, it is now appropriate to develop a little further, firstly, the aspect of production of the PAAs and of the particles which may or may not be loaded with AP and, secondly, the aspect of use of the DPs and of the suspensions of these DPs in medicinal systems for sustained and controlled release of AP.
The PAAs are obtained in a way known in itself. In this respect, reference will be made, for example, to: xe2x80x9cEncyclopedia of Polymer Science and Engineering, volume 12, p. 786; John Wiley and Sonsxe2x80x9d. In the context of the invention, use is preferably made of polymerization techniques which involve N-carboxy-xcex1-amino acid anhydrides, the preparation of which is given, for example, in: xe2x80x9cBiopolymers, 15, 1869 (1976)xe2x80x9d. Regarding the techniques for polymerizing these N-carboxy-xcex1-amino acid (NCA) monomers, details are given in the book by H. R. KRICHELDORF xe2x80x9cxcex1-Aminoacid-N-Carboxy Anhydride and Related Heterocyclesxe2x80x9d Springer Verlag (1987).
The nature of the distribution of the hydrophobic groups on the polymer chains is random or ordered, depending on the synthetic pathway chosen. In this respect, there is a multitude of reaction schemes which produce the PAAs selected as raw material for the production of the DPs according to the invention. The three reaction schemes are given hereinafter in a nonlimiting capacity:
(i) synthesis or use (step A) of a PAA copolymer comprising at least two types of hydrophobic recurrent groups, R01 and R02,
and then, by selective elimination (step B), at least one of them is eliminated so as to regenerate rAA in its unmodified state (for example, R01=methyl; R02=benzyl; step B is a step of debenzylation by hydrogenation, by addition of HBr, etc.);
(ii) synthesis or use (step A) of a hydrophobic PAA homopolymer comprising (exclusively) hydrophobic recurrent amino acids (R0-rAAs) which all bear the same hydrophobic R0 group (eg. methyl, ethyl, propyl, benzyl, stearyl),
and then the partial elimination (step B) of part of the R0 groups (eg. by virtue of hydrolysis or saponification of the poly(methyl glutamate) PAA);
(iii) synthesis or use (step A) of a hydrophilic PAA homopolymer comprising (exclusively) hydrophilic recurrent amino acids, rAAs,
and then partial hydrophobization (step B) by formation of a covalent attachment with one or more R0 groups (eg. by esterification using fatty alcohols or by ionic displacement using an alkyl halide).
In these three variants (i), (ii) and (iii) the linking unit for grafting the hydrophobic R0 groups onto the pendent side chain of the rAAs is advantageously an ester or amide function.
In practice, the hydrophobic PAA copolymers used in variant (i) are, for example, copolymers of glutamic acid and/or of aspartic acid, in which the pendent carboxylic side functions are protected by esterification according to techniques known to those skilled in the art. They may be, for example, copoly(stearyl glutamate-benzyl glutamate) obtained by polymerization of rAA01 (Glu-O-stearyl) N-carboxyanhydride and rAA02 (Glu-O-benzyl) N-carboxyanhydride.
The partial hydrophilization of the co-PAA results from the selective elimination of one of the R0 groups, by hydrolysis and/or saponification. This is equivalent to a deprotection which, in the example above, is selective debenzylation. The differences in sensitivity to cleavage of the various esters making up the hydrophobic groups R01 and R02 are exploited herein. By way of examples of known deprotection processes, mention may be made of those by saponification of methyl esters (STAHMAN et al.; J. Biol. Chem., 197, 771 (1952); KYOWA HAKKO, FR 2 152 582) or by debenzylation [BLOUT et al.; J. Amer. Chem. Soc., 80, 4631 (1958)].
With regard to variant (ii), the hydrophobic PAA homopolymer is, for example, poly(stearyl glutamate) obtained by polymerization of Glu-O-stearyl NCA. The partial hydrolysis of this hydrophobic homopolymer according to the abovementioned known deprotection methods produces a poly(glutamate)-co-(stearyl glutamate).
In variant (iii), the hydrophilic PAA homopolymer can, for example, be polyglutamic acid or a polyglutamate, synthesized by polymerization of glutamic acid NCA. The hydrophobization of this type of poly Glu PAA is carried out, for example, by reaction with stearyl iodide in basic medium. This produces poly(glutamate)-co-(stearyl glutamate). This type of technique is in particular described in [Polymer Bulletin, 32, 127 (1994)].
The formation of the DPs in liquid medium, in the form of a colloidal suspension in accordance with the invention, can be carried out during or after the synthesis of the amphiphilic PAAs defined above.
According to a preference of the invention, the formation of particles takes place by the addition of water or of salt to a solution of PAA, carrying R0 groups, dissolved in a solvent. This operation preferably consists in decreasing the solubility of the hydrophobic fraction so that it precipitates and, in so doing, causes the formation of the particles. Those skilled in the art are capable of finding other simple means for decreasing the solubility of the hydrophobic portion of the polymer, for example by modifying the temperature or the solvent(s), or by combining different techniques.
The preparation of the colloidal suspension of DPs is advantageously followed by a purification step involving techniques known to those skilled in the art, such as distillation, filtration, modification of the pH, chromatography and/or dialysis. Such methods make it possible to eliminate the unwanted salts or solvents. After this optional purification step, a colloidal suspension of DPs is obtained, which can be used directly or which can conceivably be isolated or recovered, in the context of a step IV, by any physical means which is known in itself and suitable, such as for example by filtration, by ultrafiltration, by density gradient separation, by centrifugation, by precipitation, optionally by adding salt, or by lyophilization.
According to a variant of the invention, the process for preparing the colloidal suspension is characterized in that at least one active principle is incorporated into the liquid medium containing the particles DP, so as to obtain a colloidal suspension of DPs loaded or associated with one or more active principle(s) AP.
This incorporation, which produces trapping of AP by the DPs, can be carried out in the following way:
dissolving AP in aqueous solution, and then adding the DPs, either in the form of a colloidal suspension or in the form of isolated DPs (lyophilizate or precipitate);
or adding AP, either in solution or in the pure or preformulated state, to a colloidal suspension of particles DP, optionally prepared extemporaneously by dispersing dry DPs in a suitable solvent, such as water.
The active principle, which is capable of being associated with the particles according to the invention, can be medicinal and/or nutritional. It is preferably chosen from:
proteins and/or peptides, among which the most preferably selected are: hemoglobins, cytochromes, albumins, interferons, antigens, antibodies, erythropoietin, insulin, growth hormones, factor IX, interleukins, or mixtures thereof,
polysaccharides, heparin being more particularly selected,
nucleic acids, and preferably oligonucleotides of RNA and/or of DNA,
vitamins, amino acids and trace elements,
and mixtures thereof.
The present invention is also directed toward the precursors of these particles DP consisting of the specific PAAs defined above and characterized in that they contain medicinal APs of the type of those listed above, in particular insulin, and/or APs formed by at least one vaccine, or nutritional, plant-protection or cosmetic APs.
The present invention also relates to the colloidal suspensions of DPs, characterized in that they contain the same APs as those set out above, in particular for the DP precursors.
Finally, the present invention relates to the medicinal products, or the pharmaceutical or nutritional specialties, comprising the DPs loaded with AP and defined above.
According to another of its aspects, the invention is also directed toward the use of these DPs loaded with AP, for manufacturing medicinal products such as systems for controlled release of AP.
In the case of medicinal products, they may, for example, be those which can preferably be administered orally, nasally, vaginally, ocularly, subcutaneously, intravenously, intramuscularly, intradermally, intraperitoneally, intracerebrally or parenterally.
The cosmetic applications which can be envisaged are, for example, compositions which can be applied via transdermal routes.
The plant-protection products concerned may, for example, be herbicides, pesticides, insecticides, fungicides, etc.
A subject of the present invention is also the plant-protection and cosmetic compositions comprising DPs loaded with APs of the type of those targeted above.
The examples which follow will make it possible to more clearly understand the invention in its various product/process/application aspects. These examples illustrate the preparation of polyamino acid particles which may or may not be loaded with active principles, and they also present the structural characteristics and properties of these particles.
The techniques used for the polymerization of the NCAs to polymers having block or random structures are known to those skilled in the art and are given in detail in the book by H. R. KRICHELDORF xe2x80x9cxcex1-aminoacids-N-Carboxy Anhydrides and Related Heterocyclesxe2x80x9d, Springer Verlag (1987). The following synthesis specifies the synthesis of one of them.
Synthesis of poly(GluOMe)63-poly(GluOBz)63: 10 g of NCAxe2x80x94GluOMe are solubilized in a mixture of 150 ml of dioxane and 450 ml of toluene at 60xc2x0 C. 5 ml of a solution of 0.91 g of benzylamine in 50 ml of dioxane are added to the monomer in a single amount. After 1 h, 14.1 g of NCA-GluOBz, solubilized beforehand in a mixture of 20 ml of dioxane and 60 ml of toluene, are added. The polymerization continues for a further 19-24 h. The polymer is precipitated from the reaction medium in 2 liters of methanol, to which a further 500 ml of water are added. The solid obtained is filtered, and dried in an incubator at 50xc2x0 C. under vacuum.
Yield: 90%. Composition by NMR 1 h (TFA-d) 45% molar GluOBz. Reduced viscosity (0.5% of TFA at 25xc2x0 C.) 0.3 dl/g. Molar mass by GPC: 20,000 g/mol.
10 g of polymer are then reacted at 0xc2x0 C. in 200 ml of TFA with 6 ml of trifluoromethanesulfonic acid for 15 min. During the hydrolysis of the benzyl groups, the polymer precipitates in the form of colloidal particles. After evaporation to dryness, the particles are taken up in water and neutralized at pH 7.4 with sodium hydroxide. A dialysis step ensures the elimination of the trifluoroacetate salts. The particles are finally isolated by lyophilization. Quantitative yield. Elemental analysis [Na] 8.2% (calculated composition 53% GluONa).
This polymer is prepared according to the method described in Example 1, using ethyl glutamate NCA and benzyl glutamate NCA in a 1:3 molar ratio. After the polymerization and debenzylation steps, the polymer is purified by dialysis at pH 7, and it is then lyophilized in order to obtain a white powder.
This polymer is prepared according to the method described in Example 1, using dodecyl glutamate NCA and benzyl glutamate NCA in a 1:3 molar ratio. After the polymerization and debenzylation steps, the polymer is purified by dialysis at pH 7, and it is then lyophilized in order to obtain a white powder.
10 g of polyglutamic acid (degree of polymerization 120) are dissolved in 200 ml of dimethyl sulfoxide. 15.5 g of KHCO3 are then added, followed by 2.87 ml of iodododecane. The reaction medium is maintained at 60xc2x0 C. for 40 hours under a nitrogen stream. The polymer is isolated by precipitation in 1.5 l of 0.1N hydrochloric acid, and the precipitate thus formed is washed several times in water. The desired colloidal suspension is obtained by adjusting the pH of the medium to 7.4 with sodium hydroxide, and is then dehydrated by lyophilization. The lyophilized polymer is finally washed several times in ethyl acetate, and dried under vacuum at 50xc2x0 C. Composition by NMR (TFA-d): 12% of dodecylesterified glutamic acids. Residual water after lyophilization 10.5%. % elemental analysis (calc.): C 41.29 (42.15); H 5.99 (6.01); N 7.45 (7.63); Na 10.44 (10.45).
10 mg of particles of polymer 1 are suspended in 10 ml of water or in an aqueous salt solution. This solution is then introduced into a Coulter granulometer (or laser diffractometer). The results of the particle size analysis of the various products tested are given in Table 1 below.
A solution of human insulin titrated at 1.4 mg/ml corresponding to 40 IU/ml is prepared using an isotonic phosphate buffer solution of pH 7.4. 10 mg of the DP prepared in Example 1 is dispersed in 1 ml of this insulin solution. After incubation for 15 hours at room temperature, the DP-associated insulin and the free insulin are separated by centrifugation (16,000 g, 1 hour) and ultrafiltration (filtration threshold 300,000 D). The free insulin recovered in the filtrate is assayed by HPLC or ELISA, and the amount of associated insulin is deduced therefrom by difference. The amount of DP-associated insulin is greater than 0.77 mg, which represents more than 55% of the total amount of insulin used.
The table below gives the results of the measurements of degree of association carried out on various DPs. The degree of association expresses the percentage of insulin associated, with respect to the insulin used, in a preparation titrated at 1.4 mg/ml of insulin and 10 mg/ml of DPs.
By way of comparison, insulin does not associate with sodium polyglutamate. This preliminary test is used to determine the optimum mixture of DP and of insulin in order to obtain an optimum formulation of DP and of insulin with a high degree of loading.
A formulation is prepared using DPs and insulin, the amounts of each being determined according to the degree of association measurements in Example 6.
A solution of human insulin titrated at 2.8 mg/ml corresponding to 80 IU/ml is prepared using an isotonic phosphate buffer solution of pH 7.4. 56 mg of the DP prepared in Example 1 are dispersed in 1 ml of this insulin solution. The pH and the isotonicity are adjusted, if necessary, in order to obtain a formulation at pH 7.4 and 280-300 mOs.
This preparation is diluted in increasing volumes of an isotonic 0.5% bovine albumin solution buffered at pH 7.4 with a 0.01 M phosphate buffer. The released insulin fraction is assayed by HPLC or ELISA. See attached figure. The released fraction increases with the dilution. All of the insulin is released on dilution 20. The assays by HPLC and ELISA show that  greater than 90% of the insulin is associated in this formulation.
A formulation is prepared using DPs and insulin, the amounts of each being determined according to the degree of association measurements in Example 6.
A group of 4 beagle dogs (males and females) weighing between 10 and 12 kg are fasted for 18 hours. A preparation is formulated according to Example 7 and consists of 80 IU of insulin and 56 mg of DPs (prepared according to Example 1) in 1 ml of PBS buffer. The dogs then receive a subcutaneous administration of this insulin preparation in a proportion of 1 IU/kg of weight. Blood is sampled for a glucose assay and an insulin assay before (xe2x88x922 h, xe2x88x921 h and 0 h) and after (1 h, 2 h, 4 h, 6 h, 12 h, 16 h, 20 h, 24 h, 28 h, 32 h, 36 h, 40 h, 44 h, 48 h) injection. The glucose concentrations are measured in the samples using the glucose oxidase method, and the serum insulin is 25 assayed using a radioimmunological method.

Particles based on polyamino-acid(s) and methods for preparing same


A bike carrier that includes an extension support having a vehicle support end and a second end as well as a side rail structure supported by the extension support. The side rail structure has a first section extending to a first side of the extension support and a second section extending to a second side of the extension support, and the first and second sections are arranged for supporting a respective wheel of a bike when in use. The carrier device also includes a first wheel hold down device positioned on the first side of the extension support and an intermediate support device supported by the extension support (e.g. either directly or indirectly such as via an intermediate side rail section) so as to contact a frame component of the bike when in use. The intermediate support device has an upper frame contact section that is positioned, when is use, for contact with the lower sloping frame component of the bike which extends to the hub typically. The upper contact section includes a sloped neck section which includes a frame reception recess of a slope generally corresponding to that of the sloped frame section received in the recess. A frame hold down device such as a strap is threaded through the neck section for wrapping around the sloped frame section of the bike. The intermediate support device is also preferably adjustable in height and fixed in position by the position adjustment device at a desired bike frame contact height. In addition to the intermediate frame contact device and the first wheel hold down device (which is preferably in the form of a cradle slideable along the rail structure) a second wheel hold down device, such as an independent strap, is provided for the second wheel. The invention also features a modular stacking arrangement of the same.

The present invention relates to a carrier device, which, in one preferred embodiment, is a bike carrier that is suited for attachment to a vehicle. The present invention further features a modular carrier structure that is interconnectable with other carrier structures to form a carrier assembly with the modular carrier structures preferably being arranged in a vertically stacked or staggered arrangement.
A plurality of bike carrier designs have been developed including compact strap designs that position a bike on the upper surface of a bumper and rely on strap, buckle and trunk hook components to secure the bike to the body of the vehicle. Examples of this type of bike carrier can be seen in U.S. Pat. Nos. 5,118,018 and 5,038,980. In addition to the potential damage to the vehicle created by the hook and buckle components, bike carriers of this type are generally limited to single bike support and also often involve a great deal of set up and adjustment. One advantage of this type of carrier is, however, that they can be made rather compact for shipping and retail shelf space requirements.
There also exists in the art carriers for bikes and other items that include components that attach directly to a vehicle body such as the roof of the vehicle. An example of this type of carrier can be seen in U.S. Pat. No. 5,884,824. These type of carriers are applied on a more permanent basis as compared to the hook and strap arrangements described above and thus are typically not easy to remove and instal and also introduce the added possibility of damage to the vehicle body.
The prior art also features bike carriers that are designed to extend out away from the rear end of a vehicle from an attachment point in the bumper region of the vehicle such as from a square hitch common on many vehicles, particularly sports utility vehicles or SUV""s. Many of these bicycle carriers include carriers with a relatively lengthy vertical main support bar from the top of which extends horizontal support beams that are provided with clamps or the like to attach to a bicycle generally in an upper region of its frame. Examples of this type of bicycle carrier can be seen in U.S. Pat. Nos. 5,476,202; 5,469,997 and 5,373,978. As having to attach clamping components, typically fall bike weight supporting clamping components, to the more susceptible painted frame regions of bicycles is undesirable, carriers that attach to a rear end of a vehicle also include carriers that are not designed for attachment to the upper or internal frame structure of a bicycle or other wheeled appliance, but are designed for providing an underlying platform on which the bicycle can be positioned. Prior art examples of this latter type of bicycle carrier include, for example, U.S. Pat. Nos. 5,025,932; 5,377,886; 5,497,927; and 5,570,825. While these type of carriers avoid the drawback of having attachments extending near or in contact with the bicycle""s upper, interior frame structure, they generally suffer from a plurality of other problems, such as being (1) highly complex in design; (2) not very versatile from the standpoint of being able to accommodate for variations in the desired number of bikes (e.g., motorbikes or bicycles); (3) not able to accommodate a large number of bikes; not well suited for ease in loading (e.g. difficult wheel or fork latching or the positioning of handle bars of multiple bikes at a common level, conflicting relationship); (4) bulky to the extent of presenting difficulties from the standpoint of, for example, shipping, retailer shelf space requirements and high customer handling weight; and/or (5) formed of a large number of unique components making for high manufacturing and purchase costs as well as difficulty in customer assembly.
The present invention is directed at providing a carrier device that is directed at avoiding or alleviating the various problems described above as being associated with the prior art. The present invention features a carrier device, which, in one preferred embodiment, is a bike carrier that is suited for attachment to a rear end of a vehicle such as by way of a hitch, tow ball platform or bumper attachment. A preferred embodiment of the present invention features a modular carrier structure that is interconnectable with other carrier structures to form a carrier assembly with the modular carrier structures preferably being arranged in a vertically stacked or staggered arrangement as they extend out away from the rear end of a vehicle. The vertical stacking arrangement provides the advantage of being able to retain an underlying bike support relationship while avoiding having the clearance level of the rearmost bike supporter subject to scraping during certain vehicle travel conditions (e.g., steep driveway aprons or rough terrain such as that often faced during off-road travel). Thus, because of this modular stacking arrangement, a large number of bikes can be carried (e.g., 4) while avoiding scraping problems. In addition, the vertical stacking arrangement positions the handle bars of many bike at alternate levels so as to avoid loading and unloading conflicts. Loading and unloading conflicts are also easily avoided under the design of the present invention by facilitating an alternate placement of each bike on each carrier.
A preferred embodiment of the present invention also features a modular arrangement wherein a user/customer can choose the number of individual carriers (preferably each a self-contained one bike carrier) desired. For example, a single person may wish to purchase just a single, self contained modular carrier structure and not incur the expense of a multi-support carrier and yet also wish to keep the option open for later expansion. A couple on the other hand, may prefer to purchase two modular carrier structures (one attached to the vehicle and the second attached to the one attached to the vehicle), and yet also keep open the option for later expansion without having to discard or sell the earlier purchased modular carrier structures. Furthermore, with the modular nature of the present invention an expanding family can add on to a prior number of purchased carriers based on the exact number needed. Also, because a preferred embodiment of the invention features modular carrier structures that are the same in all respects or have the exact same design (or at least the same design for all extensive purposes) there is avoided confusion both at the manufacturing level (e.g. which product is being shipped out) and at the customer level (avoiding having to return items because the purchased item was not the one intended). In addition, the preferred modular carrier support of the present invention features a highly symmetrical arrangement wherein all or essentially all of the components attached to a central element are in same design pairs and mutually useable on either side (making for high efficiency manufacturing and also ease in customer assembly once purchased).
The present invention also has the advantage of being able to have a compact break down arrangement which is beneficial from the shipping container and storage requirements (particularly from the standpoint of a retailer shelf storage requirement which is at a premium). The arrangement of the present invention is also able to provide the numerous advantages described above while avoiding the introduction of a large number of components, particularly a large number of different designed components, which again avoids many of the manufacturing, assembly and use difficulties associated with the prior art.
The present invention also provides a design that allows for rapid loading and unloading of one or a large number of bikes (e.g., 4) while still providing high stability and position maintenance assurance to the user. This is particularly true following the initial sizing of the carrier wheel cradles (which in itself is easy to achieve under the present invention) wherein the user can simply insert and remove the bike from the preadjusted cradle(s) without having to further manipulate the preset cradle position. A preferred embodiment of the invention also includes an easy to perform tie down assembly which relies on the underlying modular carrier structure(s), preferably eye-loop bolts joining together components of the carrier) for quick attachment of a single tie down strap while avoiding the placement of any buckles or the like that can lead to scratching of a bike""s painted outer coating.
An alternate arrangement of the present invention features a modular carrier with a first wheel hold or placement device at one end, an intermediate support device and preferably, a second wheel placement device. One embodiment of the invention features a cradle as the second wheel placement device; a hold down strap device as the first wheel placement device and an intermediate support device comprising a support post which supports a bike frame contact member preferably in the form of an angled support neck that has a low friction abrasion angle cap such as a plastic angle cap supported within the correspondingly angled support neck. For further tie down enhancement, the angled support neck comprises tie down strap reception slots and a tie down strap threaded therethrough for tie strap wrapping about the typically lower, sloped frame bar of a bike. Moreover, the bike frame contact member is preferably made adjustable with respect to the support post such as by way of a bolt/flat plate combination. The support post is preferably releasably secured in position on the remainder of the bike carrier such as by way of a bottom base plate with a bolt aperture for receiving the bolt which secures a side rail to the carrier extension support.
One of the preferred embodiments of the present invention features a carrier device comprising a first modular carrier structure having an extension support which includes, at a first end, means for connection with a vehicle and first modular member connection means, and having at a second end second modular member connection means. The modular carrier structure also preferably includes means for supporting an object to be carried such as a rail assembly for below support of the wheels of a bike or the like. The means for supporting an object such as a bike includes a first brace connected with said extension support and a side rail structure to which the brace is attached. The first brace is preferably a first bridge brace having a central portion connected to said extension support and more external sections connected to an under surface of said side rail structure. In addition, in one embodiment of the invention the means for supporting includes a second bridge brace having a central portion connected to said extension support and more external sections connected to an upper surface of said side rail structure. Further, in a preferred embodiment of the invention, a common bolt extends through the bottom base plate of the support post of the intermediate support device, the corresponding external section of the upper bridge brace, through the side rail structure, and then through a corresponding external section of the lower bridge brace.
Each bridge brace is, in one embodiment, a centralized bridge brace structure in that said external sections thereof each extend less than 4 inches out from an adjacent most side section of said extension support. With this type of bridge structure the side rail structure includes a first side rail and a second side rail, with the first side rail having an internal end juxtaposed to said extension support and sandwiched between above and below positioned external sections of said first and second bridge braces, and said second side rail has an internal end juxtaposed to said extension support and sandwiched between above and below positioned external sections of said first and second bridge braces. The first and second side rails also have a common design and are in contact or within xe2x85x9 of an inch of contact with respect to said extension support. Preferably the upper and lower bridge braces have a common design. The extension support also preferably includes a set of aligned holes and said bridge braces include apertures positioned for alignment with said set of aligned holes in said extension support, and said carrier device further comprising a connector that extends through each hole of said set of aligned holes and within each of said apertures of said bridge braces. The support post of the intermediate support device can thus be easily switched to one side or the other of the extension support by choosing which side rail/bridge brace combination to use as the support platform for the support post.
A central bolt/nut combination is suitable for the purposes of the present invention, and each of said side rails including a set of aligned side rail holes at respective internal ends, and the external sections of said bridge braces have apertures which are positioned for alignment with vertically aligned side rail holes of a corresponding side rail sandwiched therebetween, and said carrier device further comprises connectors such as bolt/nut combinations which extend through respective vertically aligned sets of the side rail holes and within the apertures of corresponding above and below bridge braces. As noted above, in the embodiment featuring a vertical, intermediate support device for contact with a bike component other than the wheels such as contact with the underside of the typically lower sloped frame section of a bike (extending from the handle bar support post to the crank case hub), the base plate of the support post can be secured to the upper bridge brace""s external wing section by the same connector that extends through holes in the side rail to secure the bridge brace and support post to the side rail/extension support combination.
A carrier device of the present invention also includes a first bike holding member and a second bike holding member supported by respective side rails to opposite sides of said extension support, and wherein said first bike holder is adjustably supported on one of said side rails and free to slidably adjust from an external end of said one of said support rails to said bridge braces. The first bike holder is free to slide along said one of said support rails from a free end thereof to a location at least 4 inches from said extension support without obstruction with respect to the bike carrier in a fully assembled state. The second bike holder is adjustably supported on an oppositely positioned one of said two side rails so as to be freely slidable from a free end of said oppositely positioned one of said two side rails to a location at least 4 inches from said extension section without obstruction.
In one embodiment of the invention, each of said bike holding members preferably includes a V-shaped bar cradle having a pair of legs extending off from a corresponding pair of base extensions, and said base extensions being supported by a sliding member designed for side rail sliding contact, and said holding members including a fix-in-position device for precluding movement of said sliding member, and said V-shaped bar section extending at a greater angle outward of a vertical plane than that assumed by said base extensions. An alternate embodiment of the invention, which is particularly suited for use with a bike hold down means that includes the aforementioned intermediate support device, features a first wheel placement device in the form of a flexible hold down device such as a strap with strap locking means such as a buckle, clasp, or VELCRO(trademark), hook and loop fastener. The flexible first wheel placement device is particularly suited for use with the V-shaped, angled neck support of the intermediate support device as the latter provides a degree of lateral support like a wheel cradle, and yet, the strap is easy to position and fix in the desired position in a wrapping arrangement about the wheel of a bike.
An alternate embodiment of the invention features a second modular carrier structure having an extension support which includes, at a first end, means for connection with a vehicle and first modular member connection means, and having at a second end second modular member connection means. The second modular carrier structure also includes means for supporting an object to be carried. The second modular carrier structure is connected, through use of its first modular member connecting means and the second modular connection means of said first modular carrier structure with the first modular carrier structure in a vertically offset arrangement wherein the extension support of said second modular carrier structure has an upper surface above an upper surface of the extension support of said first modular carrier structure. Preferably the lower surface of the second modular member is flush with an upper surface of the first modular carrier structure when assembled together.
The extension support of each of said first and second modular carrier structures is preferably a tubular member and the means for connection to a vehicle for each of said first and second modular carrier structures includes a first set of aligned holes, and said first modular member connection means of both said first and second modular carrier structures includes a respective set of aligned holes, and said second modular member connection means of both said first and second modular carrier structures also includes a respective set of aligned holes, and a connector is designed to extend through the aligned holes of the first modular connection means of the second modular carrier structure and through the below positioned aligned holes of the second modular member connection means of said first modular carrier structure. Also the first and second modular carrier structures are interchangeable and of the same design with respect to which one connects to the vehicle and which one connects to the modular carrier structure connected to the vehicle. In one embodiment of the present invention the connector connecting the first and second modular carrier structures also extends through the central region of the bridge braces of the first (below) carrier structure directly supported by the vehicle. In view of the multi-purpose functioning of this connector (e.g., a fastener for the upper and lower bridge braces of the first carrier structure as well as the upper and lower arranged stacked modular carrier structures, the connector is sized rather substantially (e.g., a 5xe2x85x9xc3x973xc2xdxe2x80x3 bolt with corresponding washer and nut). When a single modular carrier device is all that is needed then the aforementioned connector is used only to secure the bridge brace(s) to the extension support. In this way, a connector is readily provided when switching from a single modular carrier structure mode to a multi-carrier structure mode.
In addition, under the present invention, it is preferable to have bikes stacked in an alternating front-back relationship in going out from the first supported bike closest to the vehicle. The arrangement of the present invention is well suited for handling this alternate positioning of a plurality of bikes. For example, in the bike hold down means of the present invention, there is included a first wheel placement device (such as a hold down strap, an intermediate support post device and a cradle) with each being readily releasable for relocation (or readily initially positionable) to assume the desired location. That is, the strap and cradle are easily positionable on either of the opposite side rails and to essentially any position along the side rail length by simple manipulation of the respective locking device. Also, the intermediate support device with support post and height adjustable angle neck can easily be switched from one side rail to another simply by fastening its base to the desired bridge brace wing extension through use of the corresponding wing extension connector. In this way the first centralized support post is positioned to one side of the double stack modular carrier structures while a second is positioned to the opposite side as the first. The same thus being true with respect to the straps and cradles.
A carrier device is also featured in the present invention which includes a third modular carrier structure of a common design as said first and second modular carrier structures and said third modular carrier structure being arranged so as to be vertically offset from said second modular carrier structure upon said third modular carrier structure being interconnected with said second modular carrier structure. In addition, when using an intermediate support device, the first and third modular carrier structure would have the same intermediate extension support position, while the second modular carrier structure would have an opposite side intermediate support device position. A fourth modular unit can also be added to the third in the same manner as described above with respect to the second modular carrier structure to the first with the fourth having the same intermediate support device position on the second. In this way the carrier structures can be attached such that said second modular carrier structure assumes a stacked, vertically offset relationship with respect to said first modular carrier and so on outward depending on the number of carriers added on.
The invention also features a carrier device for supporting bikes that comprises a first modular carrier structure having a bike support platform, a second modular carrier structure having a bike support platform, and means for releasable interconnection of a forward end of said second modular carrier structure to a rear end of said first modular carrier structure such that said second modular carrier structure is positioned vertically above and offset with respect to said first modular carrier structure.
Also, the first and second modular carrier structures preferably each include centrally positioned extension supports, and the extension support of the second modular carrier structure is laid on top of a rear portion of the extension support of said first modular carrier structure, and said means for interconnecting includes a connection member which extends through each extension support so as to lock said extension supports in a stacked relationship.
The present invention also features a bike carrier device that comprises an extension support, and a side rail structure supported by said central extension support and having side rail sections extending to opposite sides of said extension support and each of said side rail sections being in contact with said extension support. There is included a pair of wheel cradle assemblies positioned on said side rail structure on opposite sides of said extension support and at least one of said cradles being adapted for slidable adjustment on a respective, supporting side rail section. Also, at least one of said wheel cradle assemblies including a releasable fix-in-position device for fixing a corresponding one of said wheel cradle assemblies in a desired position on said side rail structure. The releasable fix-in-position device also includes a wheel hold down device which includes a flexible member pinched between said cradle and a supporting one of said slide rail upon said fix-in-position device being in a locked state. The wheel hold down device also includes a flexible strap with loop adjustment and securement means that enables the strap to be opened, wrapped around a wheel and secured in a hold down state. In one embodiment, the wheel hold down device is a strap used alone to hold one wheel down in place of a second cradle while an opposite wheel cradle is still used on the other wheel. In addition, there is preferably provided a more centralized vertical post with bike frame securement member (preferably in the form of an angled V-shaped neck with a corresponding V-shaped plastic frame protection insert and a corresponding (additional) frame hold down strap). The flexible member in each embodiment, is preferably a nylon strap with securement means for fixing one section of the strap to another section so as to form a wheel locking loop. Preferably, the pinching cradle structure includes a V-shaped section and a pair of base sections, and said V-shaped section extends from vertical at a first angle between 30 and 60 degrees and said base sections extend at 0 to 10 degrees with respect to a vertical plane.
Thus, one embodiment of the present invention features a bike carrier that includes an extension support having a vehicle support end and a second end as well as a side rail structure supported by the extension support. The side rail structure has a first section extending to a first side of the extension support and a second section extending to a second side of the extension support, and the first and second sections are arranged for supporting a respective wheel of a bike when in use. The carrier device also includes a first wheel hold down device positioned on the first side of the extension support and an intermediate support device supported by the extension support (e.g. either directly or indirectly such as via an intermediate side rail section) so as to contact a frame component of the bike when in use. The intermediate support device has an upper frame contact section that is positioned, when is use, for contact with the lower sloping frame component of the bike which extends to the hub typically. The upper contact section includes a sloped neck section which includes a frame reception recess of a slope generally (within, for example, 10 degrees) corresponding to that of the sloped frame section received in the recess. A frame hold down device such as a strap is threaded through the neck section for wrapping around the sloped frame section of the bike. The intermediate support device is also preferably adjustable in height by way of a position adjustment device that allows the upper contact section to be adjusted with respect to a lower support post section of the intermediate support section and fixed in position by said position adjustment device at a desired bike frame contact height. In addition to the intermediate frame contact device and the first wheel hold down device (which is preferably in the form of a cradle slideable along the rail structure) a second wheel hold down device such as an independent strap is provided for the second wheel with the strap being readily wrapped and removed following the securement of the frame and either the prior sliding of a wheel into the cradle or the sliding of the cradle to the wheel after frame attachment. The invention also feature a bike carrier assembly having first and second modular carriers which have the same structure or substantially the same and are readily interchangeable between a direct vehicle support position or an indirect vehicle support position wherein the non-directly support modular carrier structure is staggered upwardly and secured to the back of the free end of the direct vehicle supported carrier structure. In one embodiment a common connector connects the first and second modular carrier structures while also securing in position a side rail supporting brace. The base of the intermediate support device is also preferably secured in position by way of a connector that is used to secure one of the side rails to one side of a bridge brace supported by the extension support. The intermediate support device can thus be easily switched to opposite sides of the extension support by use of different wing sections of the bridge brace.


A bill presentment service, wherein bills received from a plurality of billing entities are made available to a plurality of consuming entities via a plurality of consuming entity service providers, is provided under control of an operator of a payment processing network. Registration and preference data pertaining to each of the plurality of consuming entities are stored in a database, accessible to the operator of the payment processing network. A given one of the consuming entities which has switched from a first one of the consuming entity service providers to a second one of the consuming entity service providers is allowed to access the bill presentment service via the first one of the consuming entity service providers, prior to the switching, and via the second one of the consuming entity service providers, subsequent to the switching, using the stored registration and preference data. A subset of the registration and preference data pertains to the given one of the consuming entities. The given one of the consuming entities is allowed to access the bill presentment service via the second one of the consuming entity service providers without having to take any updating action as to at least a portion of the subset of the registration and preference data. Methods for a “one click” payment aspect, an aspect wherein a bill having a single biller account number associated therewith is routed from a billing entity to multiple consuming entity service providers for presentation to a consuming entity, and a transaction tracking aspect are also provided, as are corresponding apparatuses and computer program products.

U.S. Pat. No. 5,699,528 to Hogan discloses a system and method for bill delivery and payment over a communications network. In a bill delivery and payment system, users are able to access a server computer on a communications network to obtain bill information and pay bills. For example, such a communications network may be the Internet or the World Wide Web thereof. Using a personal computer, a user can access a Web site provided by the server computer to view the bill information and instruct the server computer as to the details of the bill payment. In a second embodiment, without visiting the web site, users are provided with electronic bills containing bill information in the form of electronic mail (e-mail) at their e-mail addresses. After opening an electronic bill, a user can make the bill payment by replying to the electronic bill.

Apparatus and method for bill presentment and payment