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Use of sugar phosphates, sugar phosphate analog, amino acids, amino acid analogs for modulating transaminases and/or the association of p36/ malate dehydrogenase |
The invention relates to the use of a substance being selected from the group consisting of “sugar phosphates, sugar phosphate analogs, amino acids, amino acid analogs, and mixtures of said substances” for producing a pharmaceutical composition for reducing weight and/or preventing delayed damage caused by diabetes mellitus by modulating the association p36/malate dehydrogenase and/or transaminases. |
1. A method for reducing weight of a subject and/or preventing delayed damage caused by diabetes mellitus by modulating the association p36/malate dehydrogenase and/or transaminases, comprising administering a pharmaceutical composition comprising a substance selected from the group consisting of sugar phosphates, sugar phosphate analogs, amino acids, amino acid analogs, and mixtures of said substances. 2. The method of claim 1, wherein the substance is selected from the group consisting of serine, cycloserine, valine, leucine, isoleucine, proline, methionine, cysteine, amino isobutyrate, aminooxyacetate, CHBA, fructose-1,6-bisphosphate, glycerate-2,3-bisphosphate, glycerate-3-phosphate, ribose-1,5-bisphosphate, ribulose-1,5-bisphosphate, analogs of such substances and mixtures of such substances. 3. The method of claim 1, wherein the substance is selected from the group consisting of compounds of the formula I and of mixtures of such compounds, wherein a and b may be identical or different and are 0 or 1, wherein R1=—H, C1-C18 alkyl, cycloalkyl or aryl, wherein R2=—H, C1-C18 alkyl, cycloalkyl or aryl, C1-C8 hydroxyalkyl, C1-C8 mercaptoalkyl, C1-C8 ether, C1-C8 thioether, C1-C8 aminoalkyl, with C1-C8 alkyl, cycloalkyl or phenyl, —CONHX2 or —CNHNHX2 N-substituted C1-C8 aminoalkyl, with -Hal and/or —OX1 substituted aryl, —OX1, —SX1, —COO−, —(CH2)n—COOX1 or —COOX1 with X1=—H, C1-C18 alkyl, cycloalkyl or aryl, and with n=1-8, wherein R3=—CN, —C═N—X2, —COO−, —COOX2, —CO—X2, —CO—NHX2 with X2=—H, C1-C18 alkyl, cycloalkyl or aryl, wherein R4=—H, —O—P, ═O, aryl, —NHY or —CO—NHZ with Y=—H, —CO—R (R=C1-C18 alkyl, cycloalkyl or aryl or —NHA, with A=H or C1-C18 alkyl, cycloalkyl or aryl), and Z=phenyl, naphthyl, with -Hal and/or —O-Hal and/or CAoHalm and/or —N—CO—CAoHalm and/or C1-C8 alkyl, cycloalkyl or aryl substituted phenyl or with -Hal and/or —O-Hal and/or C1-C8 alkyl, cycloalkyl or aryl substituted naphthyl (Hal=—F, —Cl, or —Br), wherein m=1-3 and o=3-m, wherein a and b correspond to the number of remaining carbon valences at C1 and C2, wherein via R3 a ring connection to C1 under elimination of X1 in R2 and X2 in R3 may be provided. 4. The method of claim 3, wherein a=1 and b=0, R1=—H, R2=—H, C1-C18 alkyl, cycloalkyl or aryl, C1-C8 hydroxyalkyl, C1-C8 mercaptoalkyl, C1-C8 aminoalkyl, N-substituted C1-C8 aminoalkyl, substituted aryl, —OX1, —SX1, R3=CN, R4=═O. 5. The method of claims 1 to 4, wherein the pharmaceutical composition is prepared for intravenous application. 6. The method of claims 1 to 4, wherein the pharmaceutical composition is prepared for an administration of a daily dose of 0.1 to 20 mg per kg body weight. 7. A pharmaceutical composition containing insulin and at least one substance according to one of claims 1 to 4, wherein the ratio of the substances to insulin (w/w) is in the range of 0.001:100 to 10:100. 8. A dietetic food containing one or more substances according to one of claims 1 to 4, wherein 0.01 to 10 weight parts of the substance are mixed to 100 weight parts food. 9. The composition of claim 7, wherein the ratio is in the range of 0.01:100 to 3:100. 10. The composition of claim 7, wherein the ratio is in the range of 0.1:100 to 0.3:100. 11. The composition of claim 7, wherein the composition is prepared for intravenous application. 12. The composition of claim 7, wherein the composition is prepared for an administration of a daily dose of 0.1 to 20 mg per kg body weight. 13. The composition of claim 7, wherein the composition is prepared for intravenous administration of a daily dose of 0.1 to 20 mg per kg body weight. 14. The food of claim 8, wherein 0.1 to 10 weight parts of the substance are mixed to 100 weight parts food. 15. The food of claim 8, wherein 1 to 3 weight parts of the substance are mixed to 100 weight parts food. 16. The method of claim 5, wherein the pharmaceutical composition is prepared for an administration of a daily dose of 0.1 to 20 mg per kg body weight. |
<SOH> BACKGROUND OF THE INVENTION <EOH>A widespread problem in affluent societies is obesity of many people caused by wrong nutrition. Obesity will lead to diverse health problems, from heart/circulation problems to orthopedic complications. There are the most various approaches for controlling obesity or for reducing weight of obese persons. Virtually all approaches have in common a purely dietetic element. This means that the person within a monitored period of time will take a reduced number of calories and reduce stored fat, if applicable supported by movement therapies. All these approaches have in common that after expiration of the monitored period of time, the person usually will return to the wrong nutrition and other life habits with the consequence of a subsequent weight gain. This has been termed the yo-yo effect. Diabetes mellitus patients have the additional health problem that in the course of the disease delayed damages in particular in the form of vascular damages will occur. These are irreversible. The reason for the vascular damages is an increased production of peroxides caused by the disease in the metabolization via the malate-aspartate shuttle. Prior Art. For instance, from the document Eigenbrodt, E. et al., Biochemical and Molecular Aspects of Selected Cancers, 2:311 ff (1996), various metabolism mechanisms in the cell are known, by means of which glycolytic hydrogen is transported from the cytosol into the mitochondria. These are the glycerol 3-phosphate shuttle, the malate-aspartate shuttle, and the citrate shuttle. In well-differentiated tissues, all three shuttles are active. The glycerol 3-phosphate shuttle is strongly affected by thyroxine. This leads to a strong increase of the energy consumption with thyroid gland hyperactivity. In tumor cells, the glycerol 3-phosphate shuttle is always switched off. Therefore, hydrogen generated within the glycolysis in the glycerin aldehyde 3-phosphate dehydrogenase reaction can be transported either via the glycerol 3-phosphate shuttle of the malate-aspartate shuttle into the mitochondria, where it is burned. In the case of the transport via the glycerol 3-phosphate shuttle, 2 moles ATP per mole hydrogen are generated. In the case of the transport via the malate-aspartate shuttle, 3 moles ATP per mole hydrogen are generated. The latter shuttle thus operates at a higher energy yield. Further, it is known that the malate dehydrogenase as well as transaminases are components of the malate-aspartate shuttle. From the document Mazurek, S. et al., J. Cell. Physiol. 167:238-250 (1996), it is known in the art that the malate dehydrogenase in a cell exists in three forms, a mitochondrial form comprising the mitochondrial isoenzyme and its forerunner, a cytosolic form and a form in association with the protein p36 (phosphoprotein 36). The latter form is a forerunner of the mitochondrial isoenzyme being held by the association with p36 in the cytosol. The association promotes the hydrogen transport via the malate-aspartate shuttle. The above findings have been obtained in the prior art regularly with reference to transformation-caused specialties of the cell metabolism in tumor tissue. Other illness references are not addressed. Technical Object of the Invention. The invention is based on the technical object to provide active ingredients, which are capable to control the metabolization of food such that obesity is prevented or reduced and delayed damages with diabetes mellitus are prevented. Basics of the Invention. For achieving said technical object, the invention teaches the use of a substance selected from the group consisting of “sugar phosphates, sugar phosphate analogs, amino acids, amino acid analogs, and mixtures of said substances” for producing a pharmaceutical composition for reducing weight and/or preventing delayed damage caused by diabetes mellitus by modulating the association p36/malate dehydrogenase and/or transaminases. The invention is based on one hand on the finding that the cited substances dissolve the association p36/malate dehydrogenase in the cytosol with the consequence that the isoenzyme migrates into the mitochondria, where it is removed from the cytosolic part of the malate-aspartate shuttle. The consequence is that the metabolization of food is driven to the glycerol 3-phosphate shuttles, which on one hand has a smaller energy yield and leads on the other hand to fewer peroxides. Consequently, a given amount of taken food is metabolized less efficiently. The treated person will thus lose weight with unchanged eating habits or will keep a reduced weight. Further, damages by diabetes mellitus—overweight persons are a group having an increased risk of falling ill—are prevented. The invention is further based on the finding that the cited substances simultaneously can inhibit transaminases, i.e. a synergetic effect occurs in the shift to the glycerol 3-phosphate shuttle. The term analogs designates compounds that can be deducted from the structures of natural amino acids or sugars, i.e. being different therefrom, effecting however the same or an even stronger modulation of the p36/malate dehydrogenase association and/or transaminase inhibition than the basic natural substance. An analog may in particular be a derivative; another not naturally occurring group can replace i.e. a naturally occurring functional group or an H atom. This relates to side chains as well as to the core structure; for instance, a nitrile group may in particular replace the carboxyl group of an amino acid. In the case of the sugar phosphate analogs, a —CN group may replace a phosphate group. It is also possible to replace several phosphate groups by one —CN group each. detailed-description description="Detailed Description" end="lead"? |
Routing of Peer-To-Peer Messages in a Communications Network |
The present invention relates to a method and an arrangement for routing of peer-to-peer messages (M1, M2, M3) in a communication network. The messages are routed between a first instance (Y) in a first node (A) and a second node (B). The method comprises inserting information (I1) in a first message (M1). The information is to the second node (B). Data identifying the first instance (Y) is inserted in an originating identification field (OI) in the first message (M1). A default value (DF) is inserted in a destination identification field (DI) in the first message. The first message (M1) is then sent from the first node (A) to the second node (B). In the second node (B), the default value (DF) is identified. The information (I1) in the first message (M1) is then routed to a second instance (Q) in the second node (B) whereby the second instance is selected out of several destination instances (Q, R, S) in the second node. When the two instances have been pointed out, the ongoing interchange between the nodes continue by using the originating and destination identification fields to find the right connected instance. |
1. A method for routing of peer-to-peer messages in a communication network between a first instance in a first node and a second instance in a second node, said method comprising the following steps: inserting information in a first message, to be forwarded from the first instance to the second node; inserting data identifying the first instance, in an originating identification field in the first message; said step further comprising the step of: inserting a default value in a destination identification field in the first message; sending the first message from the first node to the second node; identifying in the second node, the default value; and routing the information in the first message to a second instance in the second node, the second instance being selected out of several destination instances in the second node. 2. A method for routing of peer-to-peer messages according to claim 1, wherein the default value represents data identifying a second instance in the second node. 3. A method for routing of peer-to-peer messages according to claim 2 further comprising the steps of: comparing the second node, the first instance identified in the originating identification field, the address field, in the first message with a value in a mapping database; and routing the information in the first message to the second instance pointed out as destination instance in the mapping database. 4. A method for routing of peer-to-peer messages in a communication network according to claim 1, said method comprising the following steps: inserting information in a second message, to be forwarded from the second instance to the first node; inserting in the second message in the originating identification field, data identifying the second instance; inserting in the second message in the destination identification field, data identifying the first instance; sending the second message from the second node to the first node; and routing the information in the second message to the first instance. 5. A method for routing of peer-to-peer messages in a communication network according to claim 4, said method comprising the following steps: inserting information in a third message, to be forwarded from the first instance to the second node; inserting in the third message, data in the originating identification field identifying the first instance and data in the destination identification field identifying the second instance; sending the third message from the first node to the second node; and routing the information in the third message to the second instance. 6. A method for routing of peer-to-peer messages in a communication network according to claim 3, said method comprising the following steps: inserting information in a second message, to be forwarded from the second instance to the first node; inserting in the second message in the originating address field, data identifying the first instance; sending the second message from the second node to the first node; and routing the information in the second message to the first instance. 7. A method for routing of peer-to-peer messages in a communication network according to claim 3, said method comprising the following steps: inserting information in a second message, to be forwarded from the second instance to the first node; maintaining in the second message in the originating address field, the data identifying the first instance; sending the second message from the second node to the first node; and routing the information in the second message to the first instance. 8. An arrangement for routing of peer-to-peer messages in a communication network between a first instance in a first node and a second instance in a second node, said arrangement comprises: means for inserting of information in a first message, to be forwarded from the first instance to the second node; means for inserting data identifying the first instance, in an originating identification field in the first message; further comprising: means for inserting a default value in a destination identification field in the first message; means for sending the first message from the first node to the second node; means for identifying in the second node, the default value; and means for routing the information in the first message to a second instance in the second node, the second instance being selected out of several destination instances in the second node; 9. An arrangement for routing peer-to-peer messages according to claim 8, said arrangement further comprising: means for comparing in the second node, the first instance identified in an address field in the first message, with a value in a mapping data base; and and means routing the information in the first message to the second instance pointed out as destination instance in the mapping database. 10. An arrangement for routing of peer-to-peer messages in a communication network according to claim 8, said arrangement comprises: means for inserting information in a second message, to be forwarded from the second instance to the first node; means for inserting in the second message in the originating identification field, data identifying the second instance; means for inserting in the second message in the destination identification field, data identifying the first instance; means for sending the second message from the second node to the first node; means for routing the information in the second message to the first instance. 11. An arrangement for routing of peer-to-peer messages in a communication network according to claim 9, said arrangement comprises: means for inserting information in a third message, to be forwarded from the first instance to the second node; means for inserting in the third message, data in the originating identification field identifying the first instance and data in the destination identification field identifying the second instance; means for sending the third message from the first node to the second node; means for routing the information in the third message to the second instance. 12-13. (canceled) |
<SOH> TECHNICAL FIELD OF THE INVENTION <EOH>The present invention relates to methods and arrangements for routing of messages in a communication network. The messages are routed to one instance among several instances of a protocol implementation in the communication network. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention solves the problem with load sharing between multiple instances in nodes in a communication network. Multiple instances enables parallel processing of messages sent between the nodes. To be able to perform an effective parallel processing, a relation between the peer-to-peer instances involved in the information exchange must be kept during the whole ongoing transaction. The problem is solved by the invention by inserting information, i.e. adding information or by changing the interpretation of existing information, in the messages. The information identifies which pair of instances that are involved in the communication. More in detail, the problem is solved by a method and an arrangement for routing of messages in a communication network. The messages are routed to one instance among several instances of a protocol implementation in the communication network. The messages are sent between a first instance in a first node and a second instance in a second node. The method comprises inserting data identifying the first instance in an originating identification field in a first message. A default value is inserted in the first message in a destination identification field. The first message is then sent from the first node to the second node. The default value is identified in the second node and the first message is dispatched to a second instance in the second node whereby the second instance is selected out of several destination instances in the second node. Alternatively, data identifying the second instance in the second node is inserted already in the first node, in the destination identification field in the first message. The second instance is hereby selected in the first node out of several destination instances in the second node. The first message is then sent from the first node to the second node and the first message is routed to the second instance. In yet another alternative, only one address field is used in the message sent between the nodes. In this alternative, the method comprises inserting of data identifying the first instance in the address field. The message is then sent from the first node to the second node. The second instance is located by finding the peer instance of the first instance. This can be done e.g. by using a mapping database. When the two instances have been pointed out, the ongoing interchange between the nodes continue by using the originating and destination identification fields to find the right connected instance. An arrangement according to the invention is a mechanism that performs routing and load sharing of incoming messages to the correct instance of the receiving node whereby the load sharing and routing is based on the new inserted information. The object of the present invention is to make parallel processing possible by means of load sharing between multiple instances in nodes in a communication network. An advantage with the invention is that the implementation is scalable as regards node resources like processing power, memory size, communication links etc. Another advantage is that the node may be given extremely high communication performance and capacity. Yet another advantage is that the availability and redundancy of the communication interfaces increases since more than one instance may handle traffic. Yet another advantage is that hot upgrade of the protocol implementation is facilitated. Yet another advantage is performance/capacity licensing is facilitated (pay as you grow). |
Method of preventing adhesions wtih ifn-upsilon |
Methods for the prevention of adhesion formation and development involve the administration of therapeutic formulations to a patient which include, as active ingredients, IFN-γ or IFN-γ enhancers. The IFN-γ or IFN-γ enhancers are preferably administered to fibrosis tissues in a subject prior to an event which induces adhesion formation, such as a surgical event. |
1. A method for the prevent or remediation of surgical adhesions comprising treating a patient at risk of developing such adhesions with a therapeutic formulation containing an active ingredient selected from the group consisting of IFN-γ and IFN-γ enhancers. 2. The method of claim 1 wherein the therapeutic formulation is locally administered to the site of potential adhesion formation. 3. The method of claim 1 wherein the administration of the therapeutic formulation results in an increase in apoptosis in adhesion fibroblasts. 4. A method for the detection of a predisposition in a subject to adhesion formation which comprises the detection of elevated levels of type I collagen in the cells or tissue of a subject. 5. The method of claim 4 wherein the cells are fibroblast cells. |
<SOH> BACKGROUND OF THE INVENTION <EOH>It is well established that injuries to the peritoneal surface of the peritoneum result in the development of post-operative adhesions in the vast majority of patients following surgery. The peritoneum is composed of mesothelial cells with submesothelial tissue containing fibroblasts, macrophages and blood vessels. Cellular processes resulting in either normal peritoneal tissue repair or the development of adhesions include migration, proliferation and/or differentiation of several cell types, among them inflammatory, immune, mesothelial and fibroblast cells. Molecules produced locally by these cells regulate fibrinolytic activity, tissue remodeling and angiogenesis, as well as the synthesis and deposition of extracellular matrix material (ECM), and these processes are central to the development of adhesions. The molecular events underlying peritoneal wound healing and the development of fibrous adhesions are complex, multifactoral and not well defined. The cascade of events that leads to peritoneal wound repair in many aspects resembles the events that occur during skin wound healing, which is characterized by inflammation, cellular migration, proliferation, phenotypic differentiation and tissue remodeling. During wound healing, fibroblasts invade the wound in the first few days of healing, and these fibroblasts have multiple functions important to wound repair. These functions include collagen synthesis, ECM reorganization, and wound contraction, resulting in mature scar formation. Tissue remodeling involves the deposition and degradation of the extracellular matrix, which is a highly regulated process occurring during wound repair, and which is influenced by a host of locally expressed growth factors, cytokines and eicosanoids. The extracellular matrix is a dynamic component capable of modulating various cellular activities including cell-cell interaction, proliferation, differentiation and sequestration of potent biological response modifiers from the wound environment. In addition, excess production and deposition of the extracellular matrix is a key factor in producing tissue fibrosis throughout the body, including the development of peritoneal adhesions. Apoptosis, or programmed cell death, is a component of normal development and differentiation in most tissues, including tissues involved in adhesion development. Apoptosis is a complex process occurring in a wide variety of organisms that removes aging or injured cells from the body. This type of cell death may be inhibited by deleterious stimuli, such as hypoxia, distorting the balance of cellular proliferation, differentiation and death, thereby impairing the normal peritoneal wound repair process. Indeed, a lower degree of apoptosis is found in dermal fibroblasts isolated from the skin of keloids and hypertrophic scars, as compared to normal skin fibroblasts. There are at least two signaling pathways which are known to trigger apoptosis. The first is mediated by the interaction of membrane receptors and ligands, such as Fas ligand and TNF-α. The second pathway is triggered by exogenous stimuli such as hypoxia, radiation and chemotherapeutic drugs, wherein the death signal is transmitted through the mitochondria. This second pathway involves the altered expression of p53 and members of the Bcl-2 family. p53 point mutations have been detected in fibroblasts isolated from keloids, and the proapoptotic protein Bax which is expressed primarily in involved skin fibroblasts. In contrast to Bax, the antiapoptotic protein Bcl-2 has been found to be expressed at higher levels in fibroblast cells. Hypoxia has a variety of effects on fibroblasts, both in vivo and in vitro. Hypoxia stimulates matrix synthesis with increased expression of fibronectin, and type I and type III collagens. Hypoxia also stimulates the production of a variety of growth factors, including the profibrotic factor transforming growth factor-β1 (TGF-β1) in human mesothelial and fibroblast cells. Although fibroblasts are invariably exposed to hypoxia in ischemic conditions, the effect of hypoxia on the apoptosis of human peritoneal and adhesion fibroblasts is not known. The overexpression of TGF-β1 has been implicated in fibrotic disorders at various sites throughout the body, such as pulmonary fibrosis, glomerulonephritis, cirrhosis of the liver, and dermal scarring. Elevated levels of TGF-β expression occurs in adhesion tissues, in the peritoneal fluid of patients with adhesions, and in surgically induced adhesion formation in animal models. Mice that are heterozygous for TGF-β1 (+/−) have been shown to have significantly lower adhesions, and express at least two fold lower TGF-β1 protein in their peritoneal fluids, as compared with wild type (+/+) animals as early as 2 hours post-injury. See Krause et al., J. Invest Surg., 12, pages 31-38 (1999). Additionally, the postoperative peritoneal administration of TGF-β1 has been shown to increase the incidence of adhesion formation, while neutralizing antibodies directed against TGF-β reduce such incidence. Lucas et al., J. Surg. Res., 65, pages 135-138 (1996); and Williams et al., J. Surg. Res., 52, pages 65-70 (1991). It has been suggested that peritoneal adhesions develop in the vast majority of subjects, with more frequent occurrence in certain subjects following surgical procedures as opposed to others. The molecular basis for such predisposition is not known. Accordingly, it is an objective of this invention to provide a method for preventing or reducing the incidence of post-operative surgical adhesions with or without the use of barrier materials by addressing the molecular basis of the condition. |
<SOH> SUMMARY OF THE INVENTION <EOH>It has now been discovered that the apoptosis rate is significantly higher in human normal peritoneal fibroblasts than in adhesion fibroblasts, and that hypoxia inhibits apoptosis and enhances the proliferation of adhesion fibroblasts in vivo. These discoveries have led to the development of novel methods for treating surgical adhesions. In one aspect, the invention comprises a method for the prevention or remediation of surgical adhesions by treating a patient at risk of developing such adhesions with a therapeutic formulation that increases apoptosis of adhesion fibroblasts in vivo. Preferably, the formulations of this invention comprise IFN-γ or IFN-γ enhancers. The formulations of this invention can be applied topically, to the site of potential adhesion formation, or systemically, to increase the level of IFN-γ in tissues and cells which may be predisposed to form adhesions. While not wishing to be bound by any particular theory, it is believed that the IFN-γ may assist in preventing the overexpression and accumulation of the extracellular matrix (ECM) which is essential to peritoneal adhesion formation following surgically-induced tissue trauma. Furthermore, the IFN-γ may also assist in manipulating the ratio of active TGF-β isoforms in the extracellular matrix, and in particular, in reducing the TGF-β1/TGF-β2 ratio, or preventing activation of TGF-β isoforms. This is believed to result in a reduction in scarring and fibrosis by controlling the chemotactic recruitment of fibroblasts and inflammatory cells, by regulating angiogenesis, and by regulating the synthesis, deposition and turnover of components of the extracellular matrix. The term “IFN-γ”, as used herein, means gamma interferon, and any substances and molecules which have the biological characteristics of gamma interferon. An “IFN-γ enhancer”, as used herein, is a biological or chemical agent which increases the expression levels of IFN-γ in cells, such as peritoneal fibroblast cells. An additional aspect of this invention involves a method for the detection of a predisposition in a subject to adhesion formation which comprises the detection of elevated levels of type I collagen, but not fibronectin, in tissues in a subject. This higher level of expression of type I collagen is believed to indicate a predisposition to develop adhesions in the affected cells and tissue. Once detected, this predisposition for adhesion formation can then be treated using the treatment procedures of this invention. detailed-description description="Detailed Description" end="lead"? |
Advanced cosmetic color analysis system and methods therefor |
An advanced cosmetic color analysis system (FIG. 1) analyzes the color of a three dimensional object (element 120) to produce a plurality of color values for distinct subsets of a portion of the object, and a cosmetic analyzer (element 150) combines the color values to produce a cosmetic color determination that has relative weightings of multiple cosmetic colors. Contemplated systems are particularly useful for determination of suitable cosmetic products (or composition thereof) to achieve a particular and desired cosmetic appearance. |
1. A system comprising: a collector that captures light as a field of pixels reflecting off of a three dimensional object; a color analyzer that uses data from the pixels to produce a plurality of color values for each of a plurality of distinct subsets of the field, wherein each distinct subset comprises more than one pixel; and a cosmetic analyzer that then combines the plurality of color values from the subsets to produce a cosmetic color determination comprising relative weightings of a plurality of cosmetic colors. 2. The system of claim 1 wherein the collector is coupled to a digital video camera or a digital still camera. 3. The system of claim 1 wherein the three dimensional object is selected from the group consisting of a hair, a plurality of hair, and a portion of facial skin. 4. The system of claim 1 wherein the data from the pixels are encoded in an RGB system or a CMYK system. 5. The system of claim 4 wherein the data further comprise at least one of a hue value, a saturation value, and a luminance value. 6. The system of claim 1 wherein the plurality of color values is calculated as a frequency of predetermined color combinations. 7. The system of claim 1 wherein the plurality of distinct subsets of the field is at least ten distinct subsets. 8. The system of claim 1 wherein at least one of the plurality of distinct subsets of the field is defined by a user. 9. The system of claim 1 wherein the pixels in the plurality of distinct subsets of the field are contiguous. 10. The system of claim 1 wherein the cosmetic color determination comprising relative weightings of between and including two and four cosmetic colors. 11. The system of claim 1 wherein the cosmetic color is expressed in a descriptive cosmetic term. 12. The system of claim 1 wherein the collector is coupled to a light source that emits broad band light. 13. The system of claim 1 wherein the three dimensional object comprises hair and wherein the cosmetic color determination further comprises identification of a commercially available hair coloring product to achieve a predetermined coloration. 14. The system of claim 1 wherein the three dimensional object comprises hair and wherein the cosmetic color determination further comprises prediction of a hair color resulting from use of a commercially available hair coloring product. 15. The system of claim 1 wherein the three dimensional object comprises hair and wherein the cosmetic color determination further comprises calculation of a composition of a hair coloring product to achieve a predetermined coloration. 16. The system of claim 14 wherein the composition of the hair coloring product includes at least one color and an undertone. 17. The system of claim 1 wherein the collector is optically coupled to a magnifying device. 18. The system of claim 17 wherein the three dimensional object comprises a single hair or a strand of hair, and wherein the cosmetic color determination further comprises determination of a hair color distribution in the single hair or strand of hair. 19. The system of claim 17 wherein the cosmetic analyzer further combines the plurality of color values from the subsets to produce a hair condition determination. 20. The system of claim 19 wherein the hair condition determination includes determination of at least one of waviness, curliness, thickness, and integrity of at least one hair. 21. The system of claim 1 wherein the three dimensional object comprises hair and wherein the cosmetic color determination further comprises recommendation of a commercially available hair care product to achieve a desired outcome. 22. The system of claim 21 wherein the hair care product is a conditioner or a shampoo. 23. The system of claim 22 wherein the hair care product is commercially available from a supplier other than a supplier that provides the system. 24. The system of claim 1 further comprising a storage medium electronically coupled to the cosmetic analyzer, wherein the storage medium stores at least one of usage history and results history. 25. The system of claim 1 further comprising a storage medium electronically coupled to the cosmetic analyzer, wherein the storage medium stores personal information of a user. |
<SOH> BACKGROUND OF THE INVENTION <EOH>There are numerous systems and methods known in the art to determine color of an object, and depending on the particular application or type of object, color determination methods and systems may vary substantially. For example, many medical and other scientific applications prefer hyperspectral imaging, especially where precise measurement is necessary. Hyperspectral imaging typically provides spectral analysis (often with a resolution of less than 10 nm) of each pixel in a field, thereby generating excellent data on the object. For example, based on such imaging, tissue demarcations (such as may be present in neoplastic or other pathological conditions) may be readily identified. In contrast, all or almost all known cosmetic applications exhibit a significantly less sophisticated degree of analysis. For example, MacFarlane et al. describe in U.S. Pat. No. 6,067,504 an analytic system in which color is averaged over an entire field, rather than on a pixel-by-pixel basis. Consequently, a major drawback of such systems is that the processes of averaging frequently provide false readings where one or more hair is present having different colors. For example, one person having some dark brown hair and some white hair would be found to have the same hair color as another person having all light brown hair. Yet the cosmetic effect of the two would be entirely different. Moreover, the two persons would likely need different hair color products to achieve the same end coloration. More recent patent applications, WO 01/55956 and U.S. Ser. No. 09/493,511 (filed Nov. 3, 2000 and Jan. 28, 2000, respectively, and both incorporated by reference herein) describe analysis of subsets of a field during cosmetic color analysis of hair or skin. Such systems advantageously permit substantially increased accuracy of ‘real-life’ color using an ordinary video or other digital camera, as opposed to an expensive colorimeter. For example, the subsets can be analyzed down to a pixel-by-pixel level, which typically allows the system to compensate for shading, glare, and other effects. However, despite improved color analysis, such systems still provide output as a single color reading. Therefore, blotchy skin or highlighted areas in an otherwise homogenous hair population will nevertheless be presented as a single color output. For example, on a descriptive scale the computed color may be presented as “light blonde”, “auburn”, or “silver”. Alternatively, on a numeric scale the color may be presented as 1.78, 4.22, or 7.29. Thus, although there are various methods and systems known in the art to determine the color of an object, and particularly of a cosmetic object, all or almost all of them suffer from one or more disadvantages. Therefore there is still a need to provide improved methods and systems for cosmetic color determination. |
<SOH> SUMMARY OF THE INVENTION <EOH>The present invention is directed to methods and systems in which a collector captures light reflecting off of a three dimensional object as a field of pixels, and in which a color analyzer uses data from the pixels to produce a plurality of color values for each of various distinct subsets (comprising more than one pixel) of the field. A cosmetic analyzer combines the color values from the various subsets and produces a cosmetic color determination that includes relative weightings of a plurality of cosmetic colors. In one aspect of the inventive subject matter, the collector includes a digital video camera or a digital still camera. Therefore, it is generally preferred that the data from the pixels are RGB or CMYK encoded data (which may further include a hue, saturation, and/or luminance value). It is further preferred that the three dimensional object is a single hair, a plurality of hair, and/or a portion of facial skin. In another aspect of the inventive subject matter, the color analyzer calculates the color values as a frequency of predetermined color combinations, and it is contemplated that, among other choices, the plurality of distinct subsets of the field is at least ten distinct subsets that may or may not be defined by a user, wherein the pixels in the plurality of distinct subsets of the field are preferably contiguous. The cosmetic color determination preferably comprises relative weightings of between and including two and four cosmetic colors, wherein the cosmetic color is expressed in a descriptive cosmetic term or in one or more numeric values. In particularly preferred aspect of the inventive subject matter, the three dimensional object comprises hair and the cosmetic color determination includes an identification of a commercially available hair coloring product to achieve a predetermined coloration, or includes prediction of a hair color resulting from use of a commercially available hair coloring product. Alternatively, the cosmetic color determination may include calculation of a composition of a hair coloring product to achieve a predetermined coloration (typically comprising at least one color and at least one undertone). Where the three dimensional object comprises a single hair or a strand of hair, and where the cosmetic color determination comprises determination of a hair color distribution or determination of hair condition in the single hair or strand of hair, it is contemplated that the collector may be optically coupled to a magnifying device. In yet further contemplated aspects, the three dimensional object is hair and the cosmetic color determination comprises recommendation of a commercially available customer specific hair care product to achieve a desired outcome, wherein particularly preferred hair care products are conditioners or shampoos (which may even be commercially available from a supplier other than a supplier that provides the system). It is still further contemplated that a storage medium may be electronically coupled to the cosmetic analyzer, wherein the storage medium stores usage history, results history, and/or personal data of a user, all of which may be used to further personalize and recommend choice of suitable products. Various objects, features, aspects and advantages of the present invention will become more apparent from the following detailed description of preferred embodiments of the invention, along with the accompanying drawing. |
Double casing with netting for food products and production method therefor |
A casing is embodied as a hollow stiff cylindrical support provided with an attachment mechanism in its coupling to the stuffing horn, on which is placed a suitably shirred tube which in turn is coated with a separation sheet and finally by a tubular net, also compressed or shirred, and which together with the inner tube extends beyond the support tube and is clamped against the tube by a clip, staple or knot such that during the stuffing operation the shirred tube and the equally shirred net are axially extended, with a simultaneous yet independent extension of the two elements favored with the aid of the intermediate separation sheet. |
1-22. (cancelled). 23. A double casing for food products, comprising: a tube of an edible or inedible material; and an elastic, semi-elastic or non-elastic tubular net, the tube and the net both being shirred independently and placed coaxially, the tube being coated externally by the net which is shirred in the same extent as the tube so that it occupies an equivalent length. 24. The double casing for food products, according to claim 23, wherein the tubular net is placed farther back than the tube. 25. The double casing for food products according to claim 23, further comprising a cylindrical separation sheet made of any material that is placed between the shirred tube and the net. 26. The double casing for food products according to claim 23, further comprising an internal support made of a hollow support cylinder of a stiff material of the shirred tube. 27. The double casing for food products according to claim 26, further comprising attachment mechanism of the hollow support cylinder for attachment to a stuffing horn. 28. The double casing for food products according to claim 26 wherein the hollow support cylinder is perforated. 29. The double casing for food products according to claim 23, wherein the material of the tube is edible collagen of bovine or porcine origin. 30. The double casing for food products according to claim 23, wherein the material of the tube is inedible collagen. 31. The double casing for food products according to claim 23, wherein the material of the tube is regenerated collagen. 32. The double casing for food products according to claim 23, wherein the material of the tube is regenerated or fibrous collagen. 33. The double casing for food products according to claim 23, wherein the material of the tube is a plastic material. 34. The double casing for food products according to claim 23, wherein the material of the tube is hemp paper. 35. The double casing for food products according to claim 23, wherein the material of the tube is any type of paper. 36. The double casing for food products according to claim 23, wherein the material of the tube is a polysaccharide. 37. The double casing for food products according to claim 23, wherein the material of the tube comprises a combination of materials selected from the group consisting of edible collagen of bovine or porcine origin, inedible collagen, regenerated collagen, regenerated or fibrous collagen, a plastic material, hemp paper, any type of paper, and a polysaccharide. 38. The double casing for food products according to claim 23, further comprising a coloring pigment or mixture of pigments added to the material of the tube, continuously or discontinuously, to form a drawing, stripes, letters, numbers or any other design. 39. The double casing for food products according to claim 23, further comprising an aromatic substance added to the material of the tube. 40. The double casing for food products according to claim 23, further comprising a plastic substance added to the material of the tube. 41. The double casing for food products according to claim 23, wherein the tube is perforated in order to allow its aeration. 42. The double casing for food products according to claim 23, further comprising a clip, staple, or other attachment mechanism for holding one of the ends of the double casing. 43. A method for manufacturing a double casing with a net for food products, comprising: placing a hollow support tube inside a stick of a tube made of an edible or inedible material and shirred; placing an enveloping protection sheet around the shirred tube stick; attaching said enveloping protection to a rear portion of the hollow support tube; and shirring a tubular net concentrically to the shirred tube stick. 44. A method for manufacturing a double casing with a net for food products according to claim 43, further comprising: providing a clip, knot, staple, or other attachment mechanism for holding an end of the double casing. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The invention relates to a double casing for food products, comprised of a flexible tube of an edible or inedible material, shirred, that is externally coated with a length equivalent to the amount of the tube of a tubular net, elastic, semi-elastic or non-elastic disposed coaxially to the tube and compressed or shirred in the same extent as the tube is shirred, so that it occupies an equivalent length. The beginning of the compressed or shirred tubular net is slightly displaced from the beginning of the shirred tube, so that a few centimeters of the latter are not coated. Between the shirred tube and the compressed tubular net can be provided a cylindrical separation sheet made of any material, such as a plastic film. The above described assembly can be internally supported by a hollow cylinder made of a stiff material. The above system, continuously or discontinuously and by way of a simultaneous unshirring of the film and the net, can provide a double wrapping formed by the overlapping film and the tubular net that that can be filled by closing the film and the net jointly on the open end and stuffing inside a solid or semisolid product with a stuffing horn placed inside the support tube of the assembly. The area of shirred film not coated by the compressed or shirred tubular net ensures that the latter will not prevent the shirred film from unfolding easily because of the compression force exerted on it. As the film and the net are unshirred to form the casing, the front ends of the shirred film and net move back in the same extent, so that there is always an area of uncoated and free shirred film. The separation sheet allows the film to become unshirred uniformly and ensures that the portion of shirred film that is not coated by the compressed net is not carried away by the latter as it unfolds. The product described can be used for manual or automatic packaging of food products that may or may not be later subjected to any processes of drying, maturing, curing, cooking, sterilization, etc., in which the film casing and outer net assembly are required. The invention also relates to the method for manufacturing said product. The use of double casings for meat products, and specifically those externally including string, mesh or nets has been traditionally employed for several purposes, among which are the following: As a decorative mechanism in order to give the product an external appearance with a greater finish in some cases, or more traditional and artisan-like in others. In other cases, the object is to ensure that the finished product maintains the characteristic mark made by the net after the casing is removed. As an additional compression system for the first casing, so that air is eliminated more effectively to obtain an improved appearance and conservation of the products, or to hold tightly the stuffed product during the cooking process in order to achieve a proper bonding of the meat. For products undergoing a drying process, the mesh, string or net help the tripe remain joined to the surface of the product despite the shrinkage resulting from the drying process. As a system for holding the inner casing so that the entire stuffed piece maintains a uniform gauge. This is important for casings made of plastic, collagen or regenerated cellulose of large caliber with a tendency for their diameter to vary. Coating these pieces with semi-elastic or non-elastic nets helps solve this problem. As a system for hanging the products, whether for their heat treatment in cooking ovens or for a drying process. The external coating of string, mesh or net provides, in addition to a basic hanging system, an effective support for the weight of the product which in many cases keeps the casing from breaking. Traditionally, this second external casing is placed manually as an additional operation after the first casing is stuffed. This operation has always been labor-intensive. One of the first improvements of the traditional method is described in U.S. Pat. No. 1,505,218 (Sartore, 1924) and consists of a double casing comprising an inner cylindrical tube coated by a cylindrical net. This double casing is cut into segments of a length matching that of a piece of the final product to be stuffed. The advantages provided over the traditional method is to give the internal piece a greater resistance, so that it can be stuffed at a higher pressure, thereby eliminating the air more efficiently and reducing the number of breakages, but above all, reducing the time required to place the net after stuffing with the ensuing savings in labor costs. A second improvement is described in U.S. Pat. No. 5,024,041 (Urban, 1991), which describes a method where both the inner casing and the outer net are shirred, the former on the stuffing tube of a stuffing machine and the latter on a tube concentric to the aforementioned one having a large enough inner diameter, so that with the two closed by a staple or clip it is possible to perform the stuffing continuously and automatically. This invention also incorporates a retainer brake for the first casing. In addition to these advantages the system also allows a faster stuffing operation. U.S. Pat. No. 5,980,374 (Mercuri, 1999) provides an additional improvement, and consists of a double casing comprising an inner tubular casing and an outer elastic cylindrical net placed concentrically to each other and later shirred jointly. The advantage is that both components form a single piece and can be used as a traditional simple casing, placing them on the stuffing horn without requiring additional tubes to place the net, and further preventing the need to load said net. Although all of the aforementioned inventions constitute important improvements of the system, there remain a few significant aspects to resolve. The system disclosed by U.S. Pat. No. 5,024,041 (Urban, 1991), although providing a continuous stuffing method, requires an additional tube to support the net and a system for attaching said tube to the stuffing machine. Additionally, the net must be loaded on this tube in a separate operation, with the corresponding additional time and labor costs. Furthermore, each time the inner casing or net is finished, which are not always simultaneous, it is necessary to stop the operation to replace the exhausted supply. This also slows down the system and increases labor costs. The system described in U.S. Pat. No. 5,980,374 (Mercuri, 1999), although solving some of the aforementioned problems, such as the need for accessories to support the net, and somewhat reducing the number of stoppages for placing either the inner casing or the net, as their length is substantially the same in each unit of double shirred casing, this approach has a disadvantage in that said length is limited because of the joint shirring of the two components. Thus, in the shirred stick, the length of double casing contained in each fold is at most the distance between the elastic transversal threads of the net, as during the shirring operation and as described in the corresponding patent, the elastic transversal threads are placed spontaneously at the troughs of the folds, thereby limiting the length of the double casing contained in each one. In addition, the number of folds per unit length contained in the stick depends on the thickness of each fold, that is, the sum of the thickness of the inner casing and the thickness of the elastic thread. |
<SOH> SUMMARY OF THE INVENTION <EOH>All of these disadvantages are satisfactorily solved by the present invention, the objects of which are summarized as follows. In view of the above discussion, it is an object of the invention to provide a double casing for food products comprised of a flexible inner tube and an outer net, both shirred independently and placed coaxially, ready to be used, allowing a simultaneous application of the flexible tube and the net to any product, preferably a food product, stuffed inside it. Also the object of the invention is to provide a system that renders it unnecessary to place accessories for supporting the nets in the stuffing machines, that prevents the costly operation of loading the net in said accessories and additional stoppages in order to replace the two different packaging elements, the cylindrical casing and the net, which are not always exhausted at the same time, thereby reducing the cost of the operation. As a corollary, it can be added that the system is applicable to any stuffing machine. A further object of the invention is to provide a double casing that can contain larger amounts of tube and net in considerably shorter lengths of shirred product stick than in traditional systems, by virtue of the coaxial arrangement of the tube and the net, allowing an independent and more compact shirring thereof. Yet another object of the invention is to facilitate the joint unshirring of the tube and the net during the use of the double casing by placing a separation sheet between them. Lastly, another object of the invention is to prevent the externally shirred net from hindering the unshirring of the inner tube by placing the former farther back than the latter. |
Device and method for melting and/or vitrifying filter gas |
When fine filter dust is melted in conventional crucible or induction furnaces, a great deal of dust is generated once again. Compression of the dust is highly complex and is not always possible. According to the invention, the filter dust is collected in a dust-tight top chamber located upstream from the inlet opening of a melting aggregate that is thermally connected to a combustion chamber. The dust particles in the top chamber sink due to the effect of gravity in the melting aggregate and are melted therein. The procedure can be supported by an additional pressure gradient along the melting aggregate. The discharge of dust is largely avoided. The process is also suitable for vitrifying contaminated dust in particular. |
1-9. (canceled) 10. A device for melting and/or vitrifying filter dust in which a melting aggregate that is thermally connected to a combustion chamber and that has an inlet opening for feeding components that are to be melted and that has an outlet opening for the melted material, whereby a top chamber that can be sealed so as to be dust-tight vis-à-vis the outside atmosphere is mounted on the inlet opening of the essentially vertically positioned melting aggregate. 11. The device according to claim 10, characterized in that the melting aggregate is conical and tapered towards the outlet opening. 12. The device according to claim 11, characterized in that the top chamber is conical or funnel-shaped, whereby it is tapered towards the inlet opening. 13. The device according to claim 12, characterized in that a lock arrangement is installed upstream from the top chamber. 14. The device according to claim 10, characterized in that a lock arrangement is installed upstream from the top chamber. 15. The device according to claim 14, characterized in that a star feeder lock is provided as the lock arrangement. 16. The device according to claim 10, characterized in that the top chamber is conical or funnel-shaped, whereby it is tapered towards the inlet opening. 17. A process for melting filter dust in which substances in dust form are fed into a melting aggregate, the substances are melted by heat exposure to a heating element, that is thermally connected to the melting aggregate, and the substances are fed in liquid form to an outlet opening for purposes of further processing, whereby the substances in dust form are fed into a dust-tight top chamber mounted on the melting aggregate before the substances are melted in the melting aggregate, from where the substances sink into the melting aggregate due to the effect of gravity. 18. The process according to claim 17, characterized in that the substances are exposed to a pressure in the top chamber that is greater than the ambient pressure at the outlet opening of the melting aggregate. 19. The process according to claim 18, characterized in that the top chamber is filled with an inert gas before or during the addition of the substances in dust form. 20. The process according to claim 19, characterized in that the inert gas is nitrogen. 21. The process according to claim 19, characterized in that glass formers, for example, SiO2, are added into the top chamber for purposes of vitrifying the substances that are fed in. 22. The process according to claim 21, characterized in that glass formers are SiO2. 23. The process according to claim 17, characterized in that the top chamber is filled with an inert gas before or during the addition of the substances in dust form. 24. The process according to claim 17, characterized in that glass formers, for example, SiO2, are added into the top chamber for purposes of vitrifying the substances that are fed in. 25. The process according to claim 17, characterized in that the heating element is a combustion chamber. 26. The process according to claim 17, characterized in that the heating element is an electric heater. |
Isolation and purification procedure of vasopeptidase peptide inhibitors |
The present invention patent refers to the isolation and purification of peptides secreted by serpent venom glands, specifically Bothrops jararaca; to the peptide thus obtained, as well as to the production procedures by genetic engineering techniques in procaryotic and eukaryotic systems; to the engineered peptide thus obtained; to the production of said peptide by chemical synthesis, as well as to the peptide resulting from this chemical processing. It also refers to the utilization of said peptides, obtained by different procedures, in distinct pharmaceutical compositions, and introduced into the organism by a variety of means, in order for them to act as inhibitors of vasopeptidases, and consequently reduce systemic arterial blood pressure, and show local vasodilating action. |
1. A process for the isolation and purification of vasopeptidase peptide inhibitors, showing specificity for the carboxyl site of the angiotensin-converting enzyme, secreted by serpent venom glands (BPPs), particularly BOTHROPS JARARACA, or produced endogenously (EVASINS), having vasodilating and anti-hypertensive action, comprising of the following steps: a) providing venom, obtained from a pool of B. jararaca venom dissolved in deionized water followed by centrifugation at 1500-2000 rpm for 15 to 20 minutes thereby producing a supernatant; passing the supernatant through a gel-filtration column, equilibrated with 30-50 mM ammonium-acetate buffer, pH 5.0 to pH 6.0, at room temperature; b) conveying the sample to the top of the column and then eluting the components at a flow rate of 1.0 to 2.0 mL/min; c) obtaining the absorbance profile for each aliquot at 214 nm in order to define the constitution of the pools d) partially purifying the components of the pools showing potentiating activity by high performance liquid chromatography, for which 0.1 to 1.0% TFA (trifluoro acid)/H2O and acetonitrile/H2O (9:1) are used as solvents, and a gradient of 5% to 60% of solvent B, with a flow rate of 0.2 to 0.5 mL/min e) determining the molecular mass and the primary structure of the bradykinin potentiating peptides by mass spectrometry (ESMS-MS). 2. The process according to claim 1, characterized by the fact that in step (d) an HPLC Merck-Hitachi, model L-6200A is used, with the UV-vis detector set at 214 nm, and the reverse-phase column C-18/Beckman (5μ, 4.6×250 mm). 3-22. (canceled) 23. A vasopeptidases inhibitor with anti-hipertensive and vasodilating action, characterized by having a formula selected from the group consisting of: I pp1aa1aa2aa3P4aa5aa6P7P8, II pp1aa1aa2aa3aa4P5aa6aa7P8P9, III pp1aa1aa2aa3aa4aa5P6aa7aa8P9P10, IV pp1aa1aa2aa3aa4aa5aa6P7aa8aa9P10P11, V pp1aa1aa2aa3aa4aa5aa6aa7P8aa9aa10P11P12, and VI pp1aa1aa2aa3aa4aa5aa6aa7aa8P9aa10aa11P12P13, where: P is always proline and the remaining amino acids are always L-amino acids and are presented in the following one-letter code; pp1 is the N-terminus; aa1 is a non-basic amino acid; aa2 is a non-acid amino acid; aa3 is a non-acid amino acid; aa4 is an amino acid; aa5 is an amino acid; aa6 for formula II, aa7 for formula III, aa8 for formula IV, and aa9 for formula V is a non-basic amino acid; aa6 for formula I, aa7 for formula II, aa8 for formula III, aa9 for formula IV, and aa10 for formula V are always I or A or T wherein: G-glycine N-asparagine A-alanine Q-glutamine P-proline D-aspartic acid V-valine E-glutamic acid I-isoleucine K-lysine L-leucine R-arginine S-serine F-phenylalanine T-threonin H-histidine W-tryptophane Y-tyrosine <E-pyroglutamyl 24. The vasopeptidase inhibitor with vasodilation and anti-hypertensive action, according to claim 23, characterized by the fact that D and E are acidic amino acids, K and R are basic amino acids, and F, W, and Y are aromatic amino acids. 25. The vasopeptidase inhibitor with vasodilation and anti-hypertensive action, according to claim 23, characterized by the fact that they are peptides of 8-13 amino acids presenting a general formula, which contains the sequence motif at the carboxyl-terminus of the oligopeptide: P X1 X2 P P where X1 can be any amino acid and X2 usually is I, and the N-terminal amino acid is blocked, and P is proline. 26-33. (canceled) 34. The vasopeptidase inhibitors of claim 23 diluted in a physiogogically acceptable carrier. 35. (canceled) 36. A process for using the vasopeptidase inhibitor of claim 23 in vivo, via trans-mucosa, parenterally, or by injection, and/or intravenously for systemic action or localized action in tissue microcirculation. 37. A process for using the vasopeptidase inhibitor of claim 23 in vivo, in dosages varying between approximately 1 g to 10 mg/kg of body weight. 38. A peptide of Formula I: pp1aa1aa2aa3P4aa5aa6P7P8 (I) wherein: (a) pp1 is the N-terminus of the peptide; (b) aa1 is a non-basic amino acid; (c) aa2 is a non-acid amino acid; (d) aa3 is a non-acid amino acid; (e) P4 is proline; (f) aa5 is an amino acid selected from the group consisting of glutamine, asparagine, proline, and glycine; (g) aa6 is an amino acid selected form the group consisting of isoleucine, alanine, and threonin; (h) P7 and P8 are proline. 39. A peptide of Formula II: pp1aa1aa2aa3aa4P5aa6aa7P8P9 (II) wherein: (a) pp1 is the N-terminus of the peptide; (b) aa1 is a non-basic amino acid; (c) aa2 is a non-acid amino acid; (d) aa3 is a non-acid amino acid; (e) aa4 is an amino acid; (f) P5 is proline; (g) aa6 is a non-acid amino acid; (h) aa7 I, is an amino acid selected form the group consisting of isoleucine, alanine, and threonin; (i) P8 and P9 are proline. 40. A peptide of Formula III: pp1aa1aa2aa3aa4aa5P6aa7aa8P9P10 (III) wherein: (a) pp1 is the N-terminus of the peptide; (b) aa1 is a non-basic amino acid; (c) aa2 is a non-acid amino acid; (d) aa3 is a non-acid amino acid; (e) aa4 is an amino acid; (f) aa5 is an amino acid selected from the group consisting of glutamine, asparagine, proline, and glycine; (g) P6 is proline; (h) aa7 is a non-acid amino acid; (i) aa8 I, is an amino acid selected form the group consisting of isoleucine, alanine, and threonin; (j) P9 and P10 are proline. 41. A peptide of Formula IV: pp1aa1aa2aa3aa4aa5aa6P7aa8aa9P10P11 (IV) wherein: (a) pp1 is the N-terminus of the peptide; (b) aa1 is a non-basic amino acid; (c) aa2 is a non-acid amino acid; (d) aa3 is a non-acid amino acid; (e) aa4 is an amino acid; (f) aa5 is an amino acid selected from the group consisting of glutamine, asparagine, proline, and glycine; (g) aa6 is a non basic amino acid; (h) P7 is proline; (i) aa8 is a non-acid amino acid; (j) aa9 I, is an amino acid selected form the group consisting of isoleucine, alanine, and threonin; (k) P10 and P11 are proline. 42. A peptide of Formula V: pp1aa1aa2aa3aa4aa5aa6aa7P8aa9aa10P11P12 (V) wherein: (a) pp1 is the N-terminus of the peptide; (b) aa1 is a non-basic amino acid; (c) aa2 is a non-acid amino acid; (d) aa3 is a non-acid amino acid; (e) aa4 is an amino acid; (f) aa5 is an amino acid selected from the group consisting of glutamine, asparagine, proline, and glycine; (g) aa6 is an amino acid; (h) aa7 is a non basic amino acid; (i) P8 is proline; (j) aa9 a non-acid amino acid; (k) aa10 I, is an amino acid selected form the group consisting of isoleucine, alanine, and threonin; (l) P11 and P12 are proline. 43. A peptide of Formula VI: pp1aa1aa2aa3aa4aa5aa6aa7aa8P9aa10aa11P12P13 (VI) wherein: (a) pp1 is the N-terminus of the peptide; (b) aa1 is a non-basic amino acid; (c) aa2 is a non-acid amino acid; (d) aa3 is a non-acid amino acid; (e) aa4 is an amino acid; (f) aa5 is an amino acid selected from the group consisting of glutamine, asparagine, proline, and glycine; (g) aa6 is an amino acid; (h) aa7 is an amino acid; (i) aa8 is a non basic amino acid; (j) P9 is proline; (k) aa10 is an amino acid selected from the group consisting of glutamine, asparagine, proline, and glycine; (l) aa11 I, is an amino acid selected form the group consisting of isoleucine, alanine, and threonin; (m) P12 and P13 are proline. 44. The peptide of any one of claims 38 through 43 wherein pp1 is pyroglutamyl. 45. The peptide of any one of claims 38 through 43 wherein aa1 is an amino acid selected from the group consisting of tryptophane, serine, glycine and asparagine. 46. The peptide of any one of claims 38 through 43 wherein aa2 is an amino acid selected from the group consisting of proline, glycine, tryptophane, and arginine. 47. The peptide of any one of claims 38 through 43 wherein aa3 is an amino acid selected from the group consisting of proline, alanine, arginine, and tryptophane. 48. The peptide of any one of claims 38 through 43 wherein aa4 is an amino acid selected from the group consisting of threonin, proline, glycine, histidine, arginine, tryptophane, and glutamic acid. |
Nucleic acid which is stabilized against decomposition |
The invention relates to a nucleic acid which is stabilised against decomposition by exonucleases. Said nucleic acid contains the following constituents: a) a code sequence coding for a defined protein, b) optionally, a promoter sequence controlling the expression of the code sequence, and c) at least one molecule A added to an end of the linear sequence containing the constituents a and b, said molecule being linked to a non-immobilised, volumic molecule B. |
1. Nucleic acid stabilised against decomposition by exonucleases and containing the following constituents: a) a code sequence coding for a defined peptide or protein, b) optionally, a promoter sequence controlling the expression of the code sequence, and c) at least one molecule A added to an end of the linear sequence containing the constituents a and b, said molecule being linked to a non-immobilised, volumic molecule B. 2. Nucleic acid according to claim 1, whereby both ends of the linear sequence are linked to one molecule A each. 3. Nucleic acid according to claims 1 or 2, whereby a spacer sequence is arranged between the constituents a and/or b and the molecule A or the molecules A. 4. Nucleic acid according to claims 2 or 3, whereby either each molecule A is linked to a molecule B, or whereby both molecules A are linked to a single molecule B having at least two binding sites for a molecule A. 5. Nucleic acid according to claims 1 through 4, whereby the molecule A is Biotin or Digoxigenin and the molecule B is Avidin, Streptavidin or Anti-Digoxigenin Antibody. 6. Method for producing a nucleic acid according to claims 1 through 5 with the following process steps: 1) a linear sequence containing constituents a) and optionally b) is prepared, 2) the linear sequence from step 1) is amplified with PCR, whereby at least one primer or one primer pair is applied carrying molecule A, 3) the product from step 2) is incubated with a solution containing molecule B. 7. Application of nucleic acid according to one of the claims 1 through 5 in a process for producing a protein coded by the code sequence in a cell-free protein biosynthesis system or in a cellular protein biosynthesis system. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Bioengineering and medical applications require proteins of high quality and quantity - measured on a gram and milligram scale. As far as larger proteins are concerned, classic synthesis is hardly possible and, in any event, uneconomical. One possible means of producing proteins in large volumes is genetic engineering. For this purpose, cloned DNA, coded for the required protein, is inserted into cells, particularly procaryontic cells, as foreign DNA in the form of vectors or plasmids. These cells are then cultivated, whereby the proteins coded by the foreign DNA are expressed and extracted. Although this method allows the gain of considerable amounts of protein, the measures known, in particular cloning, are still costly. Furthermore, the cells are usually only transiently transfected and only exceptionally stably immortalised. A continuous production of protein thus requires a steady supply of fresh cells, which in turn have to be produced using the above described costly measures. A further approach is the so-called cell-free in-vitro protein biosynthesis. This method applies biologically active cell extracts that are to a large extent free of the naturally occurring cellular nucleic-acid, and which are spiked with amino acids, energy-supplying substances and at least one nucleic-acid. The added nucleic-acid does the coding for the protein that is to be produced. When DNA is applied as the nucleic-acid, a DNA-dependent RNA polymerase must be present. Of course, RNA, mRNA can also be applied directly. By means of this approach it is not only possible to produce quickly and with comparably moderate costs such proteins that could also be produced genetically, but rather, it is possible to produce such proteins that are, for example, cytotoxic and thus not expressible to any considerable degree with the usual genetic engineering systems. However, in this case the manufacturer must produce the added nucleic-acid himself, a process which is then again costly by genetic engineering methods. To improve the efficiency of a protein synthesis it is often additionally desirable to introduce regulatory sequences and other sequences such as spacers, which are not naturally linked with a protein sequence. An alternative to the genetically engineered production of complete nucleic-acids applicable in cell-free protein synthesis is the so-called expressions PCR. Here the efficient introduction of regulatory sequences (as well as other sequences promoting translational efficiency) into a nucleic-acid to be produced plays a special role within the framework of amplification. To introduce such further sequences into a target nucleic-acid, it is necessary to have very long PCR primers. However, on the one hand it is costly to produce long primers while, on the other hand, their application increases the probability of generating inhomogeneous PCR products. Independent of the method used to produce nucleic-acids for cell-free protein biosynthesis, the following basic difficulty arises. Within the framework of this method of synthesis, so-called cytolysates i.e. extracts from cells, which contain the essential components and cell elements for protein synthesis, are used. However, the application of such cytolysates requires that the (exo-) nucleases naturally existing in the original cells are, as it were, transported into the lysate. These nucleases cause decomposition of the nucleic-acids produced for the protein synthesis, and thus reduce their half-life and consequently the protein exploitation. For obvious reasons this is a disturbing factor. Naturally, the same difficulty arises in the case of cellular systems. |
Bacterial adpglucose pyrophospatase method for obtaining the latter and its use in the production of assay devices and in the obtention of transgenic plants and bacteria |
Bacterial ADPglucose pyrophosphatase, method of production, use in the manufacture of testing devices and in the production of transgenic plants and bacteria. AGPPase is an enzyme that catalyzes the hydrolysis of ADPG (adenosine diphosphate glucose). The enzyme obtained from microbial extracts is used in testing devices for determining levels of ADPG, based on the G1P (glucose-1-phosphate) released by the reaction catalyzed by AGPPase. Partial amino acid sequences of the enzyme, the sequence of the gene that codes for it and the derived protein are also described. Finally, the production of transgenic plants and bacteria that overexpress the gene of AGPPase is described. The bacteria do not accumulate glycogen, whereas the plants obtained possess a high content of soluble sugars, low starch content and high resistance to high concentrations of salts and to high temperatures. |
1. A method of production of an enzyme product of microbial origin with ADPG pyrophosphatase (AGPPase) activity, characterized in that a suspension of E. coli is submitted to extraction of the protein fraction by a buffer, sonifcation of the extract, followed by a purification procedure by successive centrifugation and precipitations and adjustments both of the ph and of the ionic strength of th medium, preferably including heating of the protein at 58° C. and cooling in ice, and purification by gel filtration, isoelectric focusing, ion exchange or other equivalent methods of purification of proteins extracted from bacteria. 2. The method as claimed in claim 1 that comprises the following steps: (1) Culture of E. coli BL21 in 30 liters of M9 minimum medium with 5 mMolar of glucose, (2) sedimentation of the bacteria by means of centrifugation, (3) resuspension in Tris-HCl 50 mM, pH 7.5 and MgC12 5 mM, (4) sonifcation, (5) ultracentrifugation at 100,000 g, (6) heating of the supernatant for 10 minutes at a temperature of 58° C., followed by cooling in ice, (7) centrifugation and concentration of the protein of the supernatant by precipitation in ammonium sulfate and resuspension in Tris-HCl 50 mM pH 7.5 and (8) purification by gel filtration chromatography, ion exchange or isoelectric focusing. 3. The method as claimed in claim 1, characterized in that the strain of E. coli used expresses a DNA fragment, represented by SEQ. ID NO: 4, amplified by PCR from the gnomic DNA of E. coli BL21 by means of two primers obtained in their turn from the 5′ and 3′ regions, represented by SEQ ID NO: 2 and SEQ ID NO: 3 respectively, of the gene that codes for AGPPase. 4. The method of claim 1 wherein the AGPPase is obtained and purified from strains of CECT5357. 5. An enzymeproduct of prokaryotic microbial origin, designated AGPPase, obtainable according to the method of claim 1 6. An enzyme product of prokaryotic microbial origin, designated AGPPase, characterized in that it displays ADPG pyrophosphatase (EC 3.6.1.21) enzymatic activity that catalyzes the hydrolysis of ADPG in equimolar fashion to G1P and AMP, does not hydrolyze molecules with phosphomonoester bonds nor mono- di- or tri-phosphate nucleotide such as, among others, ATP, UTP, ADP, UMP or AMP, requires MgC12, is not capable of hydrolyzing bis-p-nitrophenyl-phosphate, is inhibited by molybdate and phosphorylated molecules, its activity is adversely affected by reducing and chelating agents, it is very stable at pH between 7.5 and 8.5, and, in addition to ADPG, recognizes ADPribose and ADPmanose but not other nucleotide sugars such as UDPglucose or GDPmannose, among others. 7. The enzyme product as claimed in claim 5 which does not hydrolyze, among others G1P, G6P, AMP, 3-phosophoglycerate, cAMP, nor long chain nucleic acids. 8. The enzyme product as claimed in claim 5 wherein it is inhibited by inorganic pyrophosphate and phosphate esters, such as, among others, AMP, ADP, ATP, or phosphoglycerate. 9. The enzyme product as claimed in claim 5 wherein its activity is adversely affected by, among others, β-mercaptoethanol, ADTA, reduced cysteine or ascorbate. 10. The enzyme product as claimed in claim 5 wherein it does not recognize as substrate, among others, UDPglucose, GDPglucose, adenosine, 5″-phosphosulfate or bis-p-nitrophenyl-phosphate. 11. The enzyme product as claimed in claim 5 wherein it has an apparent molecular weight determined by gel filtration of about 40-50 kDa, and exhibits a Km for ADPG of 0.15 mMolar. 12. The enzyme product as claimed in claim 5 wherein it contains in its sequence at least one polypeptide shown in SEQ ID NO: 1. 13. The enzyme product as claimed in claim 5 wherein it consists of SEQ ID NO: % or homologous sequences. 14. A method for the manufacture of assay devices and/or compositions for application in the determination of ADPG. which comprises incorporating an enzyme product as claimed in claim 5 into said device. 15. An assay device for determining ADPG. which includes the enzyme product of claim 5 in such a way that determination is based on G1P released during a reaction catalyzed by AGPPase. 16. The assay device as claimed in claim 15, characterized in that determination is based on the G1P released, which is submitted to the enzyme phosphoglucomutase to produce G6P, which is in its turn reacted in coupled fashion with NAD+, by the action of the enzyme G6P dehydrogenase, obtaining 6-phosphogluconate and NADH, determinable by conventional methods: spectrophotometric, fluorimetric or of some other nature. 17. DNA sequence selected from: (a) a sequence that codes for a polypeptide that includes the amino acid sequence represented by SEQ ID NO: 1 or SEQ ID NO: 5; (b) a sequence that includes the polynucleotide sequence represented by SEQ ID NO: 4; (c) complementary sequences that hybridize that the sequence defined in a) or b) and that code for an enzyme product with AGPPase activity; (d) sequences that differ from those defined in c) owing to degeneration of the genetic code. 18. A method for producing transgenic plants that overexpress the enzyme AGPPase which comprises inoculating plants with a vector containing a DNA sequence as claimed in claim 17. 19. The m method of production of transgenic plants that overexpress AGPPase, characterized in that it uses a transformation vector that contains a plasmid that includes some of the DNA sequences of claim 17. 20. The method of production of transgenic plants as claimed in claim 19, characterized in that the transformation vector is Agrobacterium tumefaciens. 21. The method of production of transgenic plants as claimed in claim 20, characterized in that the transformation vector comprises strains of Agrobacterium tumefaciens CECT5901. 22. Transgenic plants with reduced starch content and resistant to high salinity and temperatures, obtainable according to the method of claim 19 which overexpress the enzyme AGPPase. 23. Use of the DNA sequences of claim 17 in the production of transformed bacteria that overexpress AGPPase. 24. A method of obtaining transformed bacteria that overexpress AGPPase, characterized in that it uses a transformation vector, in particular a plasmid that includes some of the DNA sequences in claim 17. 25. Transformed bacteria with reduced glycogen content obtainable according to the method of claim 24, characterized in that they overexpress the enzyme AGPPase. 26. A method of feeding animals which comprises feeding them transformed bacteria of claim 25. 27. The enzyme product as claimed in claim 7 which does not hydrolyze DNA or RNA. 28. The enzyme product as claimed in claim 8 wherein said phosphate ester is selected from the group consisting of AMP, ADP, ATP, phosphoglycerate and mixtures of two or more thereof. 29. A fish feed comprising transformed bacteria as claimed in claim 25. 30. An assay device for determining ADPG. which includes the enzyme product of claim 6 in such a way that determination is based on G1P released during a reaction catalyzed by AGPPase. |
<SOH> FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES <EOH>The invention relates to the field of the production, purification and characterization of the enzyme ADPglucose pyrophosphatase (AGPPase), its use for making devices (kits) for determining ADPG in both plant and microbial extracts, and the production of transgenic plants and bacteria in which there is overexpression of the gene that codes for AGPPase, aspP, giving rise to plants and bacteria with reduced content of starch or glycogen, respectively, high content of soluble sugars and high resistance to salinity and to high temperatures. |
Hose clamp and closing tool |
A hose clamp for limited diameters, in the form of an open band ring having overlapping band ends, is manufactured from the band material of an insert as is used in conventional hose clamps for bridging an ear gap, such insert being typically made of steel according to DIN 1.4310 having a thickness of 0.3 mm. An opening is provided near one band end and a hook for engaging the opening is provided near the other band end. For closing this hose clamp, a gripper-type tool is used, the two jaws of which, in the closed condition, constitute an essentially uninterrupted inner surface having a diameter corresponding to the outer diameter of the closed hose clamp. |
1. A hose clamp made of an open band ring (10) and having overlapping band ends (11, 12), an opening (13) disposed in one band end (11), and a hook (14) disposed in the other band end (12) for engaging the opening (13), characterised in that the band ring (10) is made of the material of a hose clamp insert having a thickness of up to 0.4 mm. 2. The hose clamp of claim 1 wherein the material of the band ring (10) is steel according to DIN 1.4310 having a thickness of 0.2 to 0.3 mm. 3. The hose clamp of claim 1 wherein the band ring (10) has border regions (16) raised outwardly from a centre portion (15) that takes a substantial part of the band width. 4. The hose clamp of claim 3 wherein the centre portion (15) of the band ring (10) has a curved cross-section. 5. The hose clamp of claim 4 wherein the centre portion (15) of the band ring (10) has an outwardly convexly curved cross-section. 6. The hose clamp of any one of claims 3 to 5 wherein the outer overlapping band end (11) has its terminal edge (19) chamfered in the areas of the raised border regions (16). 7. The hose clamp of any preceding claim wherein the hook (14) is partially punched and pressed out of the band material. 8. The hose clamp of claim 7 wherein the hook (14) in the closed condition projects beyond the plane portion of the band ring (10) no further than the outwardly raised border regions (16). 9. The hose clamp of any preceding claim, having a diameter of up to 20 mm, preferably 3 to 13 mm, wherein the band material has a width of 5 to 12 mm, preferably 8 mm, and a thickness of at most 0.4 mm, preferably 0.3 mm. 10. A tool for closing a hose clamp which consists of an open band ring (10) having overlapping ends, an opening (13) located in one band end (11) and a hook (14) located in the other band end (12) for engaging the opening (13), specifically for closing the hose clamp of any preceding claim, characterised in that the tool (20) is formed as a pair grippers the two jaws (21, 22) of which in the closed condition form an inner surface having a diameter corresponding to the outer diameter of the closed hose clamp, wherein the inner surfaces of the jaws (21, 22) form a smooth cylindrical surface over their substantial width with enlarged border regions. 11. The tool of claim 10 wherein the jaws (21, 22) in the closed condition form a substantially uninterrupted inner surface. 12. The tool of claim 10 or claim 11 wherein the jaws are exchangeable in accordance with the diameter of the respective hose clamp. 13. The tool of any one of claims 10 to 12 wherein the tips (23, 24) of the jaws (21, 22) overlap each other in the closed condition. 14. The tool of claim 13 wherein the tips (23, 24) of the jaws (21, 22) have their widths reduced to substantially one-half. 15. The tool of any one of claims 10 to 13 wherein, in the hinge region (26), one gripper arm (25) is formed by two legs, with the other gripper arm (27) crossing between the two legs. 16. The tool of claim 15 wherein the tip of one jaw has two outer tines between which a central tine formed at the tip of the other jaw engages in the closed condition of the grippers. |
<SOH> BACKGROUND OF THE INVENTION <EOH>U.S. Pat. No. 4,312,101 discloses a hose clamp which, after having been applied to the hose/nipple connection to be sealed, is tightened by deforming an ear provided in the clamp. To prevent the hose from entering the ear during tightening and thereby being lifted off the nipple, the known hose clamp is provided with an insert made of band material which is inserted into the clamp so as to bridge the gap of the ear. While an easily deformable steel according to DIN 1.4301 having a typical thickness of 0.6 mm is used for normal hose clamps, specifically clamps of the above-described type with an ear that is to be tightened, inserts are made of an essentially thinner material, typically steel according to DIN 1.4310 with a thickness of 0.2 to 0.4 mm which has high resiliency and high stiffness, is correspondingly difficult to deform and is therefore unsuited for ear clamps. The present invention is based on the idea that a band material, which is typical with such inserts for known hose clamps, is used for manufacturing the hose clamps themselves. In this way, the same starting material is used for an additional purpose, resulting in a saving of cost. In doing so, it is principally possible that finished inserts, which are intended for use with known hose clamps and are cut and formed to the shape provided therefor, are employed for making hose clamps according to the invention. The embodiment wherein the material of the band ring is steel according to DIN 1.4310 having a thickness of 0.2 to 0.3 mm and wherein the band ring has border regions raised outwardly from a center portion that takes a substantial part of the band width is advantageous for the stiffness of the hose clamp. Having the center portion of the band ring provided with a curved cross-section or having the center portion of the band ring provided with an outwardly convexly curved cross-section provides a hose clamp with an intrinsic prestress which is not only advantageous for compensating tolerances but also ensures the hose clamp to resiliently engage the hose when the hook in inserted in the opening. Having the outer overlapping band end of the open band ring have its terminal edge chamfered in the areas of raised border regions results in the hose clamp being easy to close. Having the hook partially punched and pressed out of the band material wherein the hook in the closed condition projects beyond the plane portion of the band ring no further than the outwardly raised border regions results in a hose clamp of low profile without any parts projecting far radially outwardly. A typical field of application of the hose clamp according to the invention is where the hose clamps have a diameter of up to 20 mm, preferably 3 to 13 mm, wherein the band material has a width of 5 to 12 mm, preferably 8 mm, and a thickness of at most 0.4 mm, preferably 0.3 mm. A tool for closing a hose clamp which consists of an open band ring having overlapping ends, an opening located in one band end and a hook located in the other band end for engaging the opening, specifically for closing the hose clamp wherein the tool is formed as a pair grippers with two jaws which in the closed condition form an inner surface having a diameter corresponding to the outer diameter of the closed hose clamp and wherein the inner surfaces of the jaws form a smooth cylindrical surface over their substantial width with enlarged border regions is particularly suited for closing the hose clamp according to the invention. This tool permits performing the closure by means of a simple gripper movement with the hook engaging # the opening. Having the jaws of the tool form a substantially uninterrupted inner surface in the closed condition and having the jaws be exchangeable in accordance with the diameter of the respective hose clamp are suitable measures to ensure a uniform closure of the hose clamp to a cylindrical ring. Having the tips of the jaws overlap each other in the closed condition and having the tips of the jaws have their widths reduced to substantially one-half, ensures that the hose clamp is engaged by the tool over its entire circumference. Wherein, in the hinge region, one gripper arm is formed by two legs, with the other gripper arm crossing between the two legs or wherein the tip of one jaw has two outer tines between which a central tine formed at the tip of the other jaw engages in the closed condition of the grippers, are of advantage in that they safely prevent canting during closure, which could result in deviations of the finished, closed hose clamp from the cylindrical shape. A preferred embodiment of the invention will now be explained in more detail with reference to the drawings. |
<SOH> BRIEF DESCRIPTION OF THE DRAWINGS <EOH>FIG. 1 shows a hose clamp in the open condition, FIG. 2 shows the same hose clamp in the closed condition, FIGS. 3 a to 3 c represent cross-sections through the hose clamp, FIG. 4 illustrates an open tool with an open hose clamp inserted, FIG. 5 shows the same tool in the closed condition, and FIG. 6 shows a modified jaw structure for the tool. detailed-description description="Detailed Description" end="lead"? |
Method for the production of optically brightened polyester |
In the production of polyester from a polyol, preferably di-alcohol, and a polycarboxylic acid, preferably dicarboxylic acid, an optical brightener is added in the esterification process whose structure is free from open-chain ethylenic double bonds and as a result is chemically stable to the free acid used. |
1. A process for producing optically brightened polyester by esterifying an aliphatic and/or aromatic polycarboxylic acid and a polyol, where the esterification is carried out in the presence of an optical brightener whos structure is free from one or more open-chain ethylenic double bonds. 2. The process as claimed in claim 1, where the esterification is carried out in the presence of an optical brightener of the formulae where R1 to R8 independently of one another are hydrogen, C1-C20-alkyl or groups of the formula —COOR9, where R9 is hydrogen, C1-C20-alkyl or phenyl, or R1 to R8 are a group of the formula —SO2R10, where R10 is hydrogen, C1-C10-alkyl or C1-C10-hydroxyalkyl, and A is naphthylene, phenylene, thiophenylene or biphenylene. 3. The process as claimed in claim 1, where optically brightened polyethylene glycol terephthalate is produced. 4. The process as claimed in claim 1, where aliphatic and/or aromatic polycarboxylic acid is a dicarboxylic acid. 5. The process as claimed in claim 1, where the polyol is a diol. 6. The process as claimed in claim 2, where optically brightened polyethylene glycol terephthalate is produced. 7. The process as claimed in claim 2, where the aliphatic and/or aromatic polycarboxylic acid is a dicarboxylic acid. 8. The process as claimed claim 2, where the polyol is a diol. 9. The process as claimed in claim 3, where the aliphatic and/or aromatic polycarboxylic acid is a dicarboxylic acid. 10. The process as claimed in claim 3, where the polyol is a diol. 11. The process as claimed in claim 4, where the polyol is a diol. |
Conjugated of hydroxyalkyl starch and an active agent |
The invention relates to compounds, comprising a conjugate of hydroxyalkyl starch (HAS) and an active agent, whereby the hydroxyalkyl starch is either directly covalently bonded to the active agent, or by means of a linker. The invention further relates to methods for the production of a covalent HAS-active agent conjugate, whereby HAS and an active agent are reacted in a reaction medium, characterised in that the reaction medium is water or a mixture of water and an organic solvent, comprising at least 10 wt. % water. |
1-72. (canceled). 73. A method for preparing a conjugate comprising an hydroxyalkyl starch (HAS) covalently linked to at least one active ingredient, said method comprising: providing an oxidized HAS and at least one active ingredient, wherein said oxidized HAS includes at least one oxidized reducing end group; and reacting the oxidized HAS with at least one active ingredient in a reaction medium, wherein said reaction medium comprises at least 10%/weight of water. 74. The method according to claim 73, wherein said reaction medium is a mixture of water with an organic solvent. 75. The method according to claim 73, wherein said at least one active ingredient is selected from the group consisting of vitamins, vaccines, toxins, antibiotics, antiarrythmics, appetite suppressants, anesthetics, analgetics, antirheumatics, antiallergics, antiasthmatics, antidepressives, antidiabetics, antihistames, antihypertonics, antineoplastic compounds, alkylating agents, antimetabolites, sodium channel blockers, β-receptor blockers, selective enhancers of repolarisation, calcium antagonists, local anethetics, aminopenicillins, cephalosporins, aminocephalosporine, beta-Laktam-antibiotics, carbapeneme, aminoglyco-side, tetracycline, macrolid-antibiotics, gyrase-inhibitors, glycopeptide-antibiotics, lincomy-cine, streptogramine, everninomicine, oxazolidinone, nitroimidazole, sulfonamide, so-trimoxazol, lokal antibiotics, virustatics, antimycotics, and tuberculostatics. 76. The method according to claim 73, wherein said at least one active ingredient is selected from the group consisting of a hormone, a steroid, a lipid, a protein, an oligopeptide, a polypeptide, and a nucleic acid. 77. The method according to claim 76, wherein said nucleic acid is a D- or L-nucleic acid, such as a D-DNA, L-DNA, D-RNA or L-RNA. 78. The method according to claim 73, wherein said at least one active ingredient is selected from the group consisting of an enzyme, enzyme-inhibitor, receptor, receptor-fragment, insulin, factor VIII, factor IX, cytokine, interferon, interleukin, growth factor, peptide-antibiotic, viral coat protein, haemoglobin, erythropoietin, albumin, hTPA, antibody, antibody-fragment, single chain-antibody or a derivative thereof, a steroid hormone, a hormone derived from amino acids or a hormone derived from fatty acids, mitomycin C, cyclophosphamide, bleomycin, chlorambucil, cisplatin, ara-C, fludarabin, doxorubicin, etoposid, 5-FU, MTX, vinblastin, vincristin, vindesin, hydroxyurea, 6-MP, CCNU, adenosin, chinidin, procain amide, diisopyramid, lidocain, phenytoin, mexylethin, ajamalin, parjmalium, propafenon, atenolol, propanolol, amiodaron, sotalol, verapamil, gallopamil, diltiazem, ampicillin, amoxicillin, cefotaxime, ceftazidime, vancomycine, clindamycine, metronidazol, isoniazide, rifampicine, rifabutin, rifapentin, ethambutol, pyracinamid, streptomycin, prothionamid, and dapson. 79. The method according to claim 73, wherein said at least one active ingredient includes at least one functional group selected from the group consisting of an ε—NH2-group, an α—NH2-group, a SH group, a COOH group, and a —C(NH2)2-group, and wherein said oxidized HAS binds to said at least one functional group. 80. The method according to claim 73, wherein an oxidized reducing end group of said oxidized HAS reacts with an amino group of said at least one active ingredient to result in the formation of an amide. 81. The method according to claim 73, wherein said at least one active ingredient that is provided is bound to a linker. 82. The method according to claim 73, wherein said oxidized HAS that is provided is bound to a linker. 83. The method according to claim 73, wherein said oxidized HAS is a hydroxyethyl starch having an average molecular weight of 1 to 300 kDa. 84. The method according to claim 73, wherein said oxidized HAS is a hydroxyethyl starch having an average molecular weight of 2 to 40 kDa. 85. The method according to claim 73, wherein said oxidized HAS is a hydroxyethyl starch having a molar degree of substitution of 0.1 to 0.8 and having a ratio of C2:C6 substitution in the range from 2 to 20, for each hydroxyethyl group. 86. A conjugate comprising a HAS covalently linked to at least one active ingredient produced according to the process of any one of claims 73-85. 87. A pharmaceutical composition comprising a conjugate according to claim 86 and a pharmaceutically compatible compound selected from the group consisting of a pharmaceutical carrier and an adjuvant. 88. A method for preparing a pharmaceutical composition comprising: mixing a conjugate according to claim 86 with a pharmaceutically compatible compound selected from the group consisting of a pharmaceutical carrier and an adjuvant to thereby form said pharmaceutical composition. 89. A method of treating a neoplasm comprising administering a covalent HAS-antineoplastic active ingredient compound to a subject in need thereof. 90. The method according to claim 87, wherein said neoplasm is a tumor or metastasis thereof. 91. A method of treating a heart rhythm disease comprising administering a covalent HAS-antiarrhythmic active ingredient compound to a subject in need thereof. 92. A method of treating an infectious disease comprising administering a covalent HAS-antibiotic active ingredient compound to a subject in need thereof. 93. The method of claim 90, wherein said infectious disease is caused by pathogen causative agents selected from the group consisting of bacterial pathogens, viral pathogens, and parasitic pathogens. 94. The method of claim 90, wherein said infectious disease is caused by facultatively pathogen causative agents selected from the group consisting of mycoplasms, mycobacteria, chlamydia, and rickettsia. |
<SOH> TECHNICAL BACKGROUND <EOH>The clinical use of many active ingredients of pharmaceuticals is adversely affected by a number of problems (e.g. Delgado et al., Critical Reviews in Therapeutic Drug Carrier Systems, Vol. 9 (3, 4), (1992) pp. 249-304). Parenterally administered native proteins are subject for example to excretion by the reticuloendothelial system, the liver, the kidney and the spleen. Excretion depends on the charge of carbohydrate chains, the presence of cellular receptors for the protein and molecule shape and size. The excretion limit of the glomerular filtration of the kidney is for example approx. 67 kD. As a result of proteolytic degradation, a rapid loss of biological activity can also be observed. Proteins expressed by bacteria as well as other recombinant proteins can have an increased immunogenicity and provoke life-threatening hypersensitivity reactions. Corresponding reactions naturally prevent the medical use of these products. For this reason, research has been carried out systematically in the state of the art already since the end of the 70s on the improvement of the properties of exogenic proteins by chemical modification, in particular polymerization or coupling to macromolecular polymers. Many projects concentrated on the preparation of conjugates from proteins or other active ingredients on the one hand and polyethylene glycol (PEG) on the other (e.g. U.S. Pat. No. 4,179,337). The advantages expected from respective coupling reactions comprise improved in vivo half-life of the proteins, reduced toxicity, improved stability and improved solubility of the active ingredients (Abuchowski and Davis, Enzymes as drugs, Holcenberg and Rubberts, Publisher, pp. 367-383, John Wiley & Sons N.Y. (1981)). The process of coupling the active ingredients proved to be problematic however, as the active group of the protein was inactivated by coupling to PEG or the reactions did not provide the reaction product in a suitable yield. To achieve a specific coupling which does not adversely affect the activity of the active ingredient, active groups were introduced into PEG or the active ingredient or the compounds were coupled with a linker. For this purpose, PEG is normally provided with an active group which is subsequently covalently bound to a group of a protein capable of being coupled. Thus for example, the loss of the binding activity of antibodies and their fragments after their coupling to PEG was described (Kitamura et al., Cancer Res., Vol. 51 (1991), pp. 4310-4315; and Pedley, et al., Br. J. Cancer, Vol. 79 (1994), pp. 1126-1130). To solve this problem, Chapman et al. (Nature Biotech., Vol. 17 (1999), pp. 780-783) suggest binding PEG to certain binding regions of the antibody. The loss of activity of the coupling partner is also described in WO 95/13090. As a solution, it is suggested to activate PEG with a reactive group and to bind PEG to α-interferon in the presence of a surfactant via this reactive group. Cited as preferred reactive group is N-succinimide carbonate, which is said to form a urethane bond with the ε-amino group of lysine under the conditions named. WO 96/41813 also discloses processes for the preparation of a polymer-polypeptide conjugate in which the polymer (in particular PEG) is derivatised at a specific region and then bound to a polypeptide. An amino-oxi-acetyl group is preferably introduced into PEG and this compound is then bound to a polypeptide, in particular to IL-8, hG-CSF and IL-1. In the literature, there are thus numerous examples of corresponding conjugates; e.g. PEG-insulin conjugates in U.S. Pat. No. 4,179,337, PEG-bovine-haemoglobin conjugates in U.S. Pat. No. 4,412,989, PEG-ribonuclease conjugates and PEG superoxide dismutase conjugates in Veronese et al. Applied Biochem. Biotech., Vol. 11, 141-152 (1995), PEG-IL-2 conjugates or PEG-IFN-β conjugates in U.S. Pat. No. 4,766,106, PEG-polymyxin conjugates in WO 90/15628 and PEG-IL-2 conjugates in WO 90/07939. Some conjugates are now in clinical application. For example, the properties of the enzyme asparaginase were improved by conjugate formation with PEG, and a PEG-asparaginase conjugate is commercially available under the trademark Oncaspar® for cancer therapy. Recently, a PEG-coupled G-CSF was approved by the US Food and Drug Administration (Pegfilgastim). A large number of further pegylated products are in different phases of clinical development, for example PEG-CDP870, PEG-Dronabinol, etc. (e.g. PEG-pipeline at www.enzon.com or www.inhale.com). Not only proteins, but other compounds were also coupled to PEG and other polymers according to this scheme. WO 97/33552 and WO 97/38727 disclose for example the coupling of paclitaxel to PEG and the use of the conjugate for the treatment of tumors. The use of a PEG-camptothecin conjugate for the treatment of tumors is being studied by Enzon in phase I clinical trials. Antibiotics have also been coupled to PEG. Dowling and Russell, for example, describe the pharmacokinetics of an oxytetracyclin-PEG conjugate (J. Vet. Pharmacol. Ther., vol. 23 (2000), 107-110). In the state of the art, antibiotics have also been derivatized using other methods in order to obtain new functions. For example, a depot penicillin was produced, which is a procain-penicillin derivative, i.e. a salt of the penicillin with the procain base. This derivative has an extended activity and it is used, for example, in the therapy of Syphilis. Coupling reactions with more than two compounds have also been demonstrated. For example, WO 93/23062 discloses the preparation of a coupling product from an antibody directed against a B cell lymphoma, activated PEG and a toxin. PEG-active ingredient conjugates however do not have a natural structure for which in vivo decomposition pathways have been described. Amongst others for this reason, in addition to the PEG conjugates, other conjugates and protein polymers have been produced for solving the above-named problems. Thus, there are a number of processes for cross-linking different proteins and binding of proteins to macromolecules (e.g. summary in Wong, S. S., “Chemistry of protein conjugation and cross linking”, CRCS, Inc. (1993)). Hydroxyethyl starch (HES) is a derivative of a naturally occurring amylopectin and is broken down in the body by α-amylase. The preparation of HES-protein conjugates has already been described in the state of the art (e.g. HES-haemoglobin conjugates in DE 26 16 086, hereby incorporated by reference, or DE 26 46 854, hereby incorporated by reference). Haemoglobin is a protein which could be of great clinical importance as a blood-replacement and oxygen-carrier agent (so-called Haemoglobin-Based-Oxygen Carrier, HBOC). However, although the demand for such a product was recognized early on (e.g. Rabiner, J. Exp. Med. 126, (1967) 1127), none of the known HBOC products has as yet achieved the status of an approved drug. The natural haemoglobin consists of two α and β peptide chains which each bind a haeme as a prosthetic group. Isolated haemoglobin molecules are however very unstable and rapidly break down into the more stable α,β dimers (MW 32 kDa). The biological half-life of isolated haemoglobin in the blood circulation is approx. 1 hour, as the dimers are rapidly eliminated via the kidneys. In this process, the dimers produce nephrotoxic side effects (e.g. Bunn & Jandl, J. Exp. Med. 129, (1967) 925-934). Development work on derivatized haemoglobin molecules was therefore primarily directed towards the intramolecular cross-linking of haemoglobin, the intermolecular cross-linking to form polymeric HBOC forms and/or the coupling to polymers. The known haemoglobin conjugates are described for example in Xue and Wong (Meth. in Enzymol., 231 (1994), pp. 308-322, hereby incorporated by reference) and in DE 26 16 086 and DE 26 46 854, hereby incorporated by reference. The latter discloses processes by means of which haemoglobin is bound to HES by firstly reacting HES with sodium periodate. Dialdehydes form, to which haemoglobin is bound. On the other hand, DE 26 16 086, hereby incorporated by reference, describes the coupling of haemoglobin to HES according to a process in which firstly a cross-linking agent (e.g. Bromcyan) is bound to HES and haemoglobin is then bound to the intermediate product. HES is a substituted derivative of the carbohydrate polymer amylopectin which occurs in maize starch in a concentration of up to 95%. HES has advantageous rheological properties and currently used in the clinic as a volume-replacement agent and for haemodilution therapy (Sommermeyer et al., Krankenhauspharmazie, Vol. 8(8), (1987), pp. 271-278; and Weidler et al., Arzneim.-Forschung/Drug Res., 41, (1991) 494-498). Amylopectin consists of glucose units, wherein the main chains have α-1,4-glycosidic bonds, but α-1,6-glycosidic bonds are present at the branching sites. The physical-chemical properties of this molecule are determined essentially by the type of glycosidic bonds. Because of the branched α-1,4-glycosidic bond, helical structures form with approx. 6 glucose monomers per turn. The physico-chemical and the biochemical properties of the polymer can be modified by substitution. The introduction of a hydroxyethyl group can be achieved by alkaline hydroxyethylation. The different reactivity of the relevant hydroxyl group in the unsubstituted glucose monomer vis-à-vis the hydroxyethylation can be exploited through the reaction conditions, a limited influence on the substitution pattern is thus possible. HES is therefore essentially characterized via a molecular weight distribution and a degree of substitution. The degree of substitution can be described as DS “degree of substitution” which refers to the proportion of the substituted glucose monomers of all glucose units, or as MS (“molar substitution”), which gives the number of hydroxyethyl groups per glucose unit. HES solutions are present as polydisperse compositions in which the individual molecules differ from each other with regard to the degree of polymerization, the number and arrangement of the branching sites, as well as their substitution pattern. HES is thus a mixture of compounds with different molecular weights. Accordingly, a specific HES solution is determined by an average molecular weight using statistical variables. M n is calculated as a simple arithmetic average in relation to the number of molecules (numerical average), whilst M w , the weight average, represents the mass-related measurement variable. A selective chemical binding of proteins to HES was however hitherto prevented by the fact that the HES is not activated selectively. Thus, the protein-HES conjugates known in the state of the art result from a non-selective coupling of Bromcyan-activated HES to haemoglobin (e.g. DE 26 16 086, hereby incorporated by reference). Corresponding processes can lead to polydisperse products with non-uniform properties and potentially toxic side effects. A process was first disclosed by Hashimoto (Hashimoto et al., Kunststoffe, Kautschuk, Fasern, Vol. 9, (1992) pp. 1271-1279, hereby incorporated by reference) wherein the reducing aldehyde end group of a saccharide is selectively oxidized and a reactive ester (lactone) is obtained. On the basis of this process, WO 98/01158 discloses that haemoglobin-hydroxyethyl starch conjugates can be obtained in which haemoglobin and HES are selectively linked to each other via amide bonds between free amino groups of the haemoglobin and the reducing end group of the HES present in oxidized form. Both the processes described in Hashimoto et al. and the processes according to WO 98/01158, hereby incorporated by reference, are however based on a reaction between saccharide (HES) and protein (haemoglobin) in organic solvent. Dimethyl sulfoxide (DMSO) was in fact used in the publication. One of ordinary skill in the art is aware of the fact that many proteins are subject of a change in structure in organic solvents which is not reversed in aqueous solution. Regularly, a loss of activity occurs with the change in structure. In every case, a costly removal of the organic solvent is necessary, as even residual proportions of organic solvents may not be acceptable for the intended medical use. Even the potential danger of impurities and changes in structure of the proteins is to be excluded with regard to the intended use. The object of the present invention is thus to provide improved hydroxyalkyl starch-active ingredient conjugates and processes for their preparation which lead to biologically active conjugates which can be used in everyday clinical practice. A further object of the present invention is to provide a process for the preparation of hydroxyalkyl starch-active ingredient conjugates wherein by-products are not produced in significant quantities, as these by-products also adversely affect the subsequent purification of the product to a significant extent. This object was now surprisingly solved by compounds comprising a conjugate of hydroxyalkyl starch and an active ingredient, wherein the hydroxyalkyl starch is covalently bound to the active ingredient either directly or via a linker. Corresponding HAS-active ingredient conjugates are for example obtainable by processes, wherein HAS and an active ingredient are coupled in a reaction medium, wherein the reaction medium is water or a mixture of water with an organic solvent, which comprises at least 10 weight-% water. The invention further relates to processes for the preparation of a covalent HAS-active ingredient conjugate, wherein HAS and at least one active ingredient are coupled in an aqueous reaction medium and is characterized in that the reaction medium is water or a mixture of water with an organic solvent, which comprises at least 10 weight-% water. HAS is preferably oxidized before binding to the active ingredient, a specific oxidation of the reducing end groups being particularly preferred. Alternatively, the coupling can take place via the formation of a Schiff's base between HAS and an amine group-carrying active ingredient as intermediate product. This intermediate product is then reduced, resulting in the formation of a methylene amine group. |
<SOH> BRIEF DESCRIPTION OF THE FIGURES <EOH>FIG. 1 GPC chromatogram of the coupling reaction between ox-HES 130 kD and HSA according to process A.III; FIG. 2 GPC chromatogram of the coupling reaction between ox-HES 130 kD and HSA according to process A.IV; FIG. 3 GPC chromatogram of the coupling reaction between ox-HES 130 kD and HSA according to process A.V. and with a reaction time of 2 hours; FIG. 4 GPC chromatogram of the coupling reaction between ox-HES 130 kD and HSA, process A.V., reaction time overnight; FIG. 5 GPC chromatogram of the coupling reaction between ox-HES 10 kD and HSA according to process A.V, after 2 hours ( FIG. 5 a ) and overnight ( FIG. 5 b ); FIG. 6 GPC chromatogram of the coupling reaction between ox-HES 130 kD and HSA according to process A.VII, after 24 hours reaction time; FIG. 7 GPC chromatogram of the coupling reaction between ox-HES 130 kD and HSA according to process B.V; FIG. 8 SDS-PAGE and Western Blot of different coupling reactions between HES and HSA; FIG. 9 SDS-PAGE and Western Blot of different coupling reactions between HES and HSA; FIG. 10 reaction scheme for the preparation of a HES-DNA conjugate; FIG. 11 image of a gel showing the HES-DNA conjugate prior to and after digestion with a restriction enzyme. detailed-description description="Detailed Description" end="lead"? The present invention provides for the first time compounds comprising a conjugate of hydroxyalkyl starch and an active ingredient, wherein the hydroxyalkyl starch is covalently bound to the active ingredient either directly or via a linker. The present invention further provides HAS-active ingredient conjugates which can be prepared by processes, wherein HAS and at least one active ingredient are reacted with each other in an aqueous reaction medium. The processes are further characterized in that the reaction medium is water or a mixture of water with an organic solvent, which comprises at least 10 weight-% water. Within the context of the present invention, a chemical compound is referred to as an active ingredient if the compound is suitable to be an active component of any composition for therapeutic or diagnostic purposes. Preferably, the active ingredient is an active component of a drug, i.e. the compound in a drug formulation which achieves a physiological effect after administration to a subject. An overview of the approved drugs and their active ingredients is given in the pharmacopeia. All the active ingredients named in the pharmacopeia can be used for the preparation of the HAS-active ingredient conjugates by the process according to the invention. However, according to the present invention, the term active ingredient also comprises all compounds which, although known to be suitable for diagnostic or therapeutic use, were however not able to be used up to now for this purpose, because of the problems described above. The active ingredient is preferably a vitamin, vaccine, toxin, antibiotic (or antiinfective), antiarrhythmic, appetite suppressant, anesthetic, analgesic, antirheumatic, antiallergic, antiasthmatic, antidepressant, antidiabetic, antihistamine, antihypertonic or an antineoplastic agent. Structurally, it can be for example a hormone, steroid, lipid, protein, oligo- or polypeptide, a nucleic acid, in particular a D- or L-nucleic acid, such as a D-DNA, L-DNA, D-RNA or L-RNA. The use of proteins, peptides, D- or L-nucleic acids as HAS coupling partners is particularly preferred. The compounds prepared according to the present invention retain the activity of the active ingredient and the advantageous properties of the HAS. As further advantages, the conjugates prepared according to the process according to the invention have an improved in vivo half-life of the active ingredients, reduced toxicity, improved stability and/or improved solubility of the active ingredients. After administration, the HAS chain is shortened by the α-amylase in the plasma. Thus, the activity of the coupling product can be determined as activity of the native coupling product, i.e. directly after the coupling, or as activity of the metabolized coupling product, i.e. after in vivo metabolizing of the coupling product. In vivo metabolizing can be simulated by an in vitro degradation. The activity of the active ingredient may be determined by methods which are known for this compound in the state of the art. For example, the activity of an antineoplastic agent is determined as inhibitory concentration (IC), and the activity of an antiinfective agent is determined as minimal inhibitory concentration (MIC). Preferably, the determination is performed in vitro with appropriate target cells (e.g. Chow et al., Haematologica, Volume 86 (2001), pages 485-493, herein incorporated by reference). The in vitro effects can further be confirmed by a relevant animal model (e.g. for example the mouse model of the renal cell carcinoma described in Changnon et al., e.g. BJU Int., Volume 88 (2001), page 418-424, herein incorporated by reference). Compared to the non-coupled substance, the native coupling product can have an increased or reduced activity. Preferably, however, the activity is not reduced more than 5-fold, more preferably not more than 3- or 2-fold. The metabolized product preferably has an activity comparable to that of the non-coupled substance, i.e. prior to the coupling, the metabolized conjugate has at least 50%, preferably at least 75% of the activity of the active ingredient, wherein a retention of at least 95% of the activity is particularly preferred. In the context of the present invention, the term “hydroxyalkyl starch” is used to refer to starch derivatives which are substituted with a hydroxyalkyl group having 1 to 3 carbon atoms. Thus, the group designated as “hydroxyalkyl starch” comprises hydroxymethyl starch, hydroxyethyl starch and hydroxypropyl starch. The use of hydroxyethyl starch (HES) as a coupling partner is particularly preferred for all embodiments of the invention. According to the invention, it is preferred that the hydroxyethyl starch has an average molecular weight (weight average) of 1-300 kDa, wherein an average molecular weight of 5 to 200 kDa is particularly preferred. Furthermore, hydroxyethyl starch may have a molar degree of substitution of 0.1 to 0.8 and a ratio of C 2 :C 6 -substitution in the range of 2-20, in each case relative to the hydroxyethyl groups. For coupling the active ingredient to the HAS, it may be necessary in a first step to introduce an active group into the active ingredient and/or the HAS. Corresponding active groups can for example be thiol groups or amino groups (e.g. Examples). Further, the active ingredient and the HAS can be coupled to each other by use of a linker. Any crosslinking agent can be used as a linker. Numerous crosslinking agents such as SMCC (succinimidyl-4-(N-maleimido-methyl)cyclohexane-1-carboxylate; e.g. Example 7) are commercially available and well-known to the person skilled in the art (e.g. alphabetic list of the “cross-linking reagents” in the product catalogue of the company Perbio and www.piercenet.com ). According to a further embodiment of the present invention, water-soluble antibiotic derivatives which contain an amino sugar, in particular HAS-daunorubicin and HAS-doxorubicin conjugates, and processes for their preparation, as far as they are disclosed in DE 101 29 369, herein incorporated by reference, are not within the scope of the present invention, e.g. DE 101 29 369 is disclosed with the proviso that said disclosure is not within the scope of the present invention. According to a preferred embodiment, the present invention relates to compounds comprising a conjugate of HAS and an antineoplastic active ingredient and their use for the treatment of tumors. Among others, tumor cells differ from normal somatic cells in that tumor cells are no longer subject to a physiological growth control and therefore have an increased rate of cell division. The therapeutic use of antineoplastic active ingredients in tumor therapy is based on this difference, since the toxic activity of the antineoplastic active ingredients is primarily directed against proliferating cells. As a consequence, compounds are designated as antineoplastic active ingredients or cytostatics if they exhibit a toxic activity against proliferating cells (basics of oncology and current therapeutic approaches are for example summarized in: Internistic Oncology, Schmoll et al. (eds.), Springer, 1996). With respect to their chemistry, antineoplastic active ingredients represent a very heterogeneous group. In addition to the inhibition of proliferation, the induction of apoptosis, programmed cell death, gains importance in the discussions over the last years. A classification of the antineoplastic active ingredients can for example be performed based on the relevant target molecules (Schmoll et al., see above): 1. Compounds which inhibit DNA biosynthesis, for example as antimetabolites, such as MTX, 5-FU, Ara-C or hydroxy urea. 2. Compounds, which act on the DNA, for example by strand break induction, intercalation, modification of interstrand cross-linking, topoisomerase toxins, such as alkylating agents, platinum complexes, anthracyclins, bleomycin, actinomycin-D or epipodophyllo toxins. 3. Compounds which act on the RNA, for example by blocking mRNA-synthesis by intercalation or incorporation into the RNA, including anthracyclins, bleomycin, actinomycin-D or antimetabolites. 4. Compounds, which act on proteins, for example on the level of receptor binding (e.g. hormones or antagonists), by inhibition of tubulin polymerization (e.g. by vinca alkaloids), by protein cross-linking (for example by alkylating agents) or phosphorylation (e.g. by inhibitors of protein kinase C). Due to the antineoplastic activity, all active ingredients have considerable side effects, which primarily occur as inhibition of fast proliferating tissues. For this reason, in particular erythro-, leuko- and trombopoiesis are inhibited and the growth of mucous membrane epithelia is adversely affected. As a consequence, gastrointestinal disorders or non-reversible impairments of spermatogenesis or anovulation, respectively, can occur. The skin and the skin accessory organs are also usually affected. For example, many patients suffer from a reversible loss of hair. In severe cases, the side effects can lead to an acute loss of the kidney function and toxic-related organ damages to heart, lung, liver and nervous system. Finally, as a consequence of the immunosuppressive effect, an increased number of infections have to be expected. The preparation and investigation of conjugates which contain an antineoplastic agent were therefore focused on the improvement of the tolerance of the active ingredient. For this purpose, different antineoplastic active ingredients have been coupled to macromolecules such as dextran (e.g. Kojima et al., J. Pharm. Pharmakol., Vol. 32 (1980), p. 30-34; Nakane et al., J. Pharm. Pharmakol., vol. 40 (1988), p. 1-6, Nomura et al., J. Controlled Release, Vol. 52 (1998), p. 239-252; Sato et al., J. Pharm. Sci., Vol. 78 (1989), p. 11-16, the disclosures of which are hereby incorporated by reference). In several cases, an improved anti-tumor effect of the conjugates was demonstrated. As an alternative, active ingredients such as mitomycin C were also coupled to N-succinylchitosan (Song et al., J. Controlled Release, Vol. 42 (1996), p. 93-100; hereby incorporated by reference), carboxymethylchitin (Song et al., Arch. Pract. Pharm. Vol. 53 (1993), p. 141-147; hereby incorporated by reference) and oligopeptides (Soyez et al., J. Controlled Release, Vol. 47 (1997), p. 71-80; hereby incorporated by reference). When compared to the individual antineoplastic active ingredient, again, an improved anti-tumor activity of the conjugates was observed in the majority of analyses. According to the invention it was now surprisingly found, that HAS-active ingredient conjugates which comprise an antineoplastic active ingredient have an improved toxic effect against tumor cells and/or a reduced toxicity for other cells. Therefore, the conjugates allow for a broader therapeutic range. The plasma half-life of the conjugates is significantly increased. This allow overcomes the repair mechanisms in tumor cells by longer exposition. Simultaneously, the present invention enables slower flooding, in particular in healthy tissue, whereby a reduced peak concentration and an improved tolerance for the patient is achieved. For the preparation of the conjugates according to the invention, any antineoplastic active ingredient can be used. The antineoplastic active ingredient can, for example, be selected from the group consisting of alkylating agents, antimetabolites, antibiotics or natural substances. According to a preferred embodiment, the antineoplastic active ingredient is mitomycin C, cyclophosphamid, bleocin, chlorambucil, cisplatin, Ara-C, fludarabine, doxorubicin, etoposide, 5-FU, MTX, vinblastine, vincristine, vindesine, hydroxy urea, 6-MP or CCNU. The use of mitomycin C as active ingredient is particularly preferred. Mitomycin C belongs to the group of antibiotics and contains an aziridine group and a quinone group and a mitosane ring. The active ingredient is used for the treatment of renal cell carcinoma and bladder tumors as well as other urologic diseases. The compound gains its activity only upon metabolization in hypoxyic cells (this means preferably in tumor cells) by intracellular enzymatic or spontaneous chemical reduction of the quinone and loss of the methoxy group. Preferably, HAS can be coupled to this methoxy group via a linker. After intracellularly cleaving off the substituent, the same active ingredient is present inside the the cell which causes an alkylating cross-linking of the DNA thereby exhibiting its toxic effect. As an alternative, HAS may also be coupled to one of the two NH 2 -groups. Mitomycin C shows a typical tissue specificity. According to the invention, it is preferred that this specificity—in particular for excretory organs—is increased by HAS-coupling. According to the invention, the antineoplastic active ingredient can be coupled to HAS by use of any method. However, a specific coupling to the reducing end groups of HAS is preferred, since this procedure generates a defined conjugate. According to one embodiment of the invention, hydroxyethyl starch may be coupled to the methoxy group of mitomycin C. Coupling to the methoxy group of mitomycin C can take place via a linker. According to a further embodiment, the present invention relates to processes for the preparation of a compound comprising a conjugate of HAS and an antineoplastic active ingredient. The process comprises steps, in which HAS is covalently coupled to an antineoplastic active ingredient, either directly or via a linker, and the conjugate is isolated. Further, the invention relates to pharmaceutical compositions which comprise a compound comprising a conjugate of HAS and an antineoplastic active ingredient. The pharmaceutical composition can furthermore comprise a pharmaceutically compatible carrier and/or a cytokine. Preferably, the cytokine is IL-2, α-interferon, γ-interferon. The pharmaceutical composition can be in any application form that is known in the state of the art. For example, the composition can be formulated for oral or parenteral administration. The formulation of the composition is performed according to processes known in the state of the art. In addition to the active ingredient, the composition generally comprises a pharmaceutically compatible carrier and one or more auxiliaries and optionally preservatives, solubility promoters, etc. Finally, the present invention relates to the use of a compound comprising a conjugate of HAS and an antineoplastic active ingredient for the preparation of a medicament for the treatment of tumors and/or their metastases, in particular for the treatment of urologic tumors and/or metastases of urologic tumors, for the treatment of metastases of the renal cell carcinoma, or for the treatment of diseases of the lymphatic system, such as CLL, Hodgkin-lymphoma, NHL, multiple myeloma, Waldenström's syndrome. According to this embodiment of the invention, the medicament can further comprise a cytokine, such as IL-2, α-interferon, γ-interferon. The use of the compounds according to the invention for the preparation of a medicament for the treatment of urologic tumors and/or metastases of urologic tumors, such as for the treatment of metastases of the renal cell carcinoma is particularly preferred. Presently, a curative therapy of the renal cell carcinoma can neither be achieved with a combination chemotherapy nor with mitomycin C alone. This might be due to the unfavourable pharmacokinetics of the compound, since the portion of renal elimination only amounts to approximately 18%. Since HAS is almost completely eliminated via the kidney, the conjugate exhibits a higher percentage of renal elimination compared to the non-conjugated substance. This embodiment of the present invention utilizes the intracellular intermediate storage of HAS. In particular, highly substituted HAS species (HAS 200/0.62) show an increased intracellular storage, in the extreme case even an overload. This phenomenon has also been observed in the area of the proximal tubule (Peron et al., Clinical Nephrology, Vol. 55 (2001), p. 408-411, hereby incorporated by reference). According to this embodiment, the present invention provides an accumulation of an antineoplastic active ingredient in certain target cells or tissues. Therefore, the improved pharmacokinetics of the conjugates make it possible to achieve a considerablely higher concentration in the cells of the target organ while using low systemic concentrations. This medical use is preferably employed on the hypernephroid carcinoma and the chromophylic renal carcinoma which constitute approximately 90% of all histological types. According to an alternative embodiment, the invention relates to the use of the compounds according to the invention for the preparation of a medicament for the treatment of diseases of the lymphatic system, such as CLL, Hodgkin lymphoma, NHL, multiple myeloma, Waldenström's syndrome. By coupling of HAS to an antineoplastic active ingredient according to the invention, the intracellular uptake of the active ingredients is decelerated dependent on the chain length and the degree of substitution. Furthermore, radioactive kinetic studies have shown that HAS is stored in certain organs, among others in lymphatic organs, for a longer time than in the whole body (e.g. Bepperling et. al., Crit. Care, Vol. 3, Suppl. 1 (1999), p. 153, hereby incorporated by reference). Thus, accumulation of the conjugate in the target cells occurs which results in improved pharmcokinetics with a lower systemic toxicity. The treatment of diseases of the lymphatic system using fludarabin as an active ingredient is preferred. Fludarabin is a halogenated adenine analogue which is resistant to deamination. The invention further relates to the use of the compounds according to the invention for the preparation of a medicament for the treatment of cutaneous/local primary malignant neoplasms or their metastases. For this, two effects can be utilized, the directed increased uptake by the recited tissues and the decelerated transport of the HAS conjugates out of the tissue. Both effects lead to an accumulation of the conjugate in the target cells. The invention further relates to the use of the compounds according to the invention for the preparation of a medicament for the treatment of diseases of the hematologic system or oncologic diseases, such as non-small cell lung cancer and small cell lung cancer, breast cancer, esophagus squamous cell carcinoma, renal cell carcinoma, testicular carcinoma, malignant melanoma, ALL or CML. In particular, when using the conjugates for the treatment of the renal cell carcinoma, advantages arise due to the strong accumulation of the compound in the affected tissue by the increased hydrophilicity of the conjugate and the stronger renal elimination resulting thereof. For this embodiment of the invention, the use of vindesine as active ingredient is particularly preferred. The invention further relates to the use of the compound according to the invention for the preparation of a medicament, wherein the compound is used as a combination therapy with one or more further antineoplastic active ingredients or cytokines. The combination therapy can be performed by administration of an agent containing all active ingredients, or by administration of two or more different compositions, each of which containing one active ingredient. The present invention further provides processes for the preparation of a medicament comprising a cytokine and a compound according to the invention which is suitable for new combination therapies. Corresponding agents are in particular suitable for the treatment of the advanced renal cell carcinoma. According to another particularly preferred embodiment of the invention, conjugates of HAS and an antiarrhythmic active ingredient as well as their use for the treatment of arrhythmia are provided. Deviations from the temporary sequence and regularity of the heartbeat (arrhythmia) from the normal heart rate are referred to as arrhythmia. In the majority of cases, these deviations are caused by cardiac excitation or conduction disorders. Substances which are suitable for the treatment of arrhythmia, in particular ventricular arrhythmia, are referred to as antiarrhythmic active ingredients or antiarrhythmics. Dependent on the effect of the antiarrhythmic active ingredients it is distinguished between sodium channel blockers (quinidine, procainamide, disopyramide, etc.) beta-receptor blockers (atenolol, propanolol, etc.), selective repolarisation prolonging active ingredients (amiodarone, sotalol, etc.), calcium antagonists (verapamil, gallopamil, etc.) and local anesthetics. However, the antiarrhythmic active ingredients customary in the state of the art partially exhibit a short duration of action. For example, adenosine is an antiarrhythmic active ingredient with a very short half-life. The duration of action of this substance is only several minutes. In many cases, prolongation of half-life and duration of action is necessary. Additionally, several antiarrhythmic active ingredients have pro-arrhythmogenic side effects and partially even an increase in mortality. The present invention provides, among others, improved antiarrhythmic active ingredients which, for example, have a prolonged duration of action. According to the invention, it was surprisingly found that the HAS-antiarrhythmic conjugates have a significantly longer in vivo plasma half-life and that the activity of the active ingredients is not adversely affected to a significant extent by coupling to HAS. According to the present invention, any antiarrhythmic active ingredient can be used for the preparation of the conjugates. The active ingredient can be selected from the group consisting of sodium channel blockers, beta-receptor blockers, selective repolarization prolonging active ingredients, calcium antagonists and local anesthetics. Preferably, the active ingredient is adenosine, quinidine, procainamide, disopyramide, lidocaine, phenytoin, mexiletine, ajamaline, Parjmalium, propafenone, atenolol, propanolol, amiodarone, sotalol, verapamil, gallopamil or diltiazem, wherein the use of adenosine is particularly preferred. According to an embodiment of the present invention, coupling between the antiarrhythmic active ingredient and the HAS takes place via the reducing end groups of the HAS. When adenosine is used, this active ingredient can for example be bound to the HAS via the amino group, wherein a coupling between the amino group of the adenosine and the reducing end group of the HAS is particularly preferred. A coupling variant, wherein native adenosine is present after metabolisation (separating off the HAS) is preferred. As an alternative, the active ingredient can be coupled to the HAS via a so-called linker. The present invention further relates to pharmaceutical compositions comprising one of the compounds according to the invention. Generally, the pharmaceutical composition further comprises a pharmaceutically compatible carrier, and it can be formulated, for example, for intravenous application. Finally the invention relates to the use of the compounds according to the invention for the preparation of a medicament for the treatment of arrhythmia, in particular for the treatment of ventricular arrhythmia. According to an alternative embodiment, the invention relates to the use of a compound according to the invention for the preparation of a medicament for the induction of apoptosis, for example in tumor tissues or in inflammatory tissues. The present invention relates to compounds comprising a conjugate of HAS and an antiinfective active ingredient or an antibiotic, respectively, as well as their use for the treatment of infectious diseases. The penetration of microorganisms (viruses, bacteria, fungi, protozoa) into a macroorganism (plant, animal, human) and the propagation in this macroorganism is called infection. Formation and course of an infectious disease substantially depend on pathogenicity of the microorganism and immunity of the macroorganism. For decades, antiinfective active ingredients have been used as chemotherapeutics in order to fight infectious diseases. A. Flemings identified Penicillin already in 1928 by the active ingredient's characteristic to form staphylococci-free areas on culture plates. Penicillin was the first antibiotic that was obtained on an industrial scale, and it gained great importance in clinical practice. Today, active ingredients from a group of β-lactam antibiotics which are produced from a fungus of the species Penicillium (for example, P. chrysogenum and P. notatum ) are designated as penicillins. The bacteriocidal effect is based on blocking the synthesis of the bacterial cell wall. The penicillin inactivates the bacterial enzyme transpeptidase, thereby preventing cross-linking of the polysaccharide chains of the cell wall murein. Since the discovery, numerous active ingredients were isolated and synthesized which inhibit the growth of microorganisms or kill microorganisms. Most antibiotics originate from Streptomyces species (approximately 65%) which were isolated from soil. It is assumed that these substances are used by the microorganism to suppress competitors in the soil. The number of isolated antibiotics is estimated to be approximately 8000, approximately 100 thereof can be used in the field of medicine. A classification of the active ingredients into different substance classes was performed according to different aspects, for example chemical structure or mode of action. Meanwhile, antibiotics are approved not only for fighting infectious diseases, but also as immuno depressants, cytostatics in anti-tumor therapy, plant protectives, for the preservation of foods, as fattening auxiliary agent in the feeding of animals, etc. In recent years, numerous strains of microorganisms occurred which are resistant to antibiotics. In addition to single-resistant strains, multi-resistant strains were frequently found which complicates fighting of certain diseases. When studying the activity of different antibiotics against certain pathogens, it was found that several of the active ingredients, for example amoxycillin or ampicillin, almost exclusively act extracellularly (Scaglione et al., Chemotherapie, Vol. 39 (1993), 416-423; Balland et al., J. Antimicrob. Chemother. Vol. 37 (1996), 105-115, the disclosures of which are hereby incorporated by reference). Therefore, these active ingredients cannot be used against microorganisms which primarily are present inside the cell. Ampicillin-nanoparticles have been produced in order to improve the intracellular activity (e.g. Balland et. al., see above). For infections such as tuberculosis or other infections caused by mycobacteri, broadening of the spectrum of treatment possibilities would be desirable due to the combination therapy which is always required. In view of other intracellular infections such as chlamydia infection, for which the potential importance for the pathogenesis of arteriosclerosis was only recently discovered (Stille und Dittmann, Herz, Vol. 23 (1998), p. 185-192, hereby incorporated by reference), intracellular antibiotics with a depot effect could represent an important progress in therapy and prophylaxis. According to the invention, it was now surprisingly found that coupling of antiinfective active ingredients to HAS results in improved pharmacokinetic characteristics of the active ingredients, in particular in a prolonged in vivo half-life, an improved intracellular uptake and/or effect of the active ingredient. According to the invention, any antiinfective ingredient or antibiotic, respectively, can be used. Preferably, an active ingredient is selected from the group consisting of amino penicillins, cephalosporines, amino cephalosporines, beta-lactam-antibiotics, carbapenems, amino glycosides, tetracyclines, macrolide antibiotics, gyrase inhibitors, glycopeptide antibiotics, lincomycins, streptogramins, everninomicins, oxazolidinones, nitroimidazoles, sulfonamides, co-trimoxazol, local antibiotics, virustatics, antimycotics, tuberculostatics. It may for example be ampicillin, amoxicillin, cefotaxim, ceftazidim, vancomycin, clindamycin, metronidazol, isoniazid, rifampicin, rifabutin, rifapentin, ethambutol, pyracinamide, streptomycin, prothionamide, or dapsone, wherein the use of an amino penicillin, such as ampicillin, amoxycillin, macrolide or of streptomycin is particularly preferred. According to one embodiment of the present invention, an amino penicillin is used as an active ingredient which is directly and covalently coupled to the hydroxyethyl starch via the amino group of the amino penicillin. According to another embodiment, an amino cephalosporin is used instead of the amino penicillin, thereby achieving a reduced allergenicity. As further embodiments, macrolide-HAS couplings may be used, wherein erythromycin or a derivative thereof is used, in particular erythromycylamin. As an alternative, streptomycin can be used as active ingredient. According to a particularly preferred embodiment of the present invention, the coupling between the antiinfective active agent and the hydroxyethyl starch may take place via the reducing end groups of the hydroxyethyl starch. In accordance with a further embodiment of the present invention, the antiinfective active agent is coupled to the hydroxyethyl starch via a linker. The present invention further comprises pharmaceutical compositions, which comprise a compound according to the invention. Usually, the pharmaceutical compositions further comprise a pharmaceutically compatible carrier. Finally, the present invention relates to the use of one of the compounds according to the invention for the preparation of a medicament for the treatment of an infectious disease. The pharmaceutical composition may in particular be suitable for the treatment of infectious diseases which, amongst others, are caused by intracellular pathogens. These may originate from the complete spectrum of pathogenics and facultative pathogenics, for example bacterial, viral or parasitic pathogens, mycoplasms, mycobacteria, chlamydia, rickettsia, etc. In a further aspect of the present invention, HAS-nucleic acid conjugates are provided. Presently, nucleic acid libraries are screened in large scale for nucleic acids which have a desired activity. For example, a respective activity can be the ability of a nucleic acid to bind to certain other nucleic acids, receptors or viral proteins. This binding may be stimulated or inhibited by a biological signal. For this purpose, in addition to naturally occurring D-DNA and D-RNA molecules, also L-DNA and L-RNA molecules are used which differ from the naturally occurring molecules in that they contain L-ribose or L-deoxyribose instead of the corresponding D-forms as components of the nucleic acid (e.g. WO 98/08856, hereby incorporated by reference). In the context of the present invention, it was shown that HAS-nucleic acid conjugates can be prepared which may retain their natural function (e.g. Example 7). The present invention further provides processes for the preparation of covalent HAS-active ingredient conjugates. The processes can be performed in an aqueous or organic reaction medium, wherein carrying out the coupling in an aqueous medium is preferred. Thus, processes for the preparation of a covalent HAS-active ingredient conjugate are provided in which HAS and at least one active ingredient are reacted with each other in a reaction medium. The reaction medium is characterized in that it is water or a mixture of water with an organic solvent, which comprises at least 10 weight-% water. The reaction medium of the process according to the invention comprises at least 10 wt.-%, preferably at least 50 wt.-%, in particular at least 80 wt.-%, such as for example 90 wt.-%, or even up to 100 wt.-%, water, and accordingly up to 90 wt.-%, preferably up to 50 wt.-%, in particular up to 20 wt.-%, for example 10 wt.-%, or even up to 0 wt.-%, organic solvent. The reaction takes place in an aqueous phase. The preferred reaction medium is water. The process according to the invention is already advantageous because toxicologically unacceptable solvents need not necessarily be used, and thus, with the product prepared according to the invention, the removal of even small residues of toxicologically unacceptable solvents which is always necessary according to the known process in order to avoid the undesired contamination with solvent is dispensed with. Furthermore, the additional quality control necessary according to the process known in the art for residues of toxicologically harmful solvents can be omitted because the process according to the invention favors the use of toxicologically acceptable solvents. Solvents preferred according to the invention are for example toxicologically harmless protic solvents such as ethanol or propylene glycol. Furthermore, it is an advantage of the process according to the invention that irreversible or reversible structural changes of proteins or peptides induced by organic solvents, which cannot be systematically excluded in processes in organic solvents, are basically avoided. The product obtained with the process according to the invention is consequently different from that prepared in DMSO. According to the invention it was, furthermore, surprisingly found that a coupling of HAS to active ingredients can be carried out in an aqueous solution without secondary reactions being observed to a significant extent. The process according to the invention thus leads directly to improved products of great purity. The process according to the invention thus makes possible for the first time the simple preparation of HAS-active ingredient conjugates in which the active ingredient is present in active form and the advantageous properties of the HAS are retained. No particular processes are necessary to isolate the HAS-active ingredient conjugate from the reaction mixture as the reaction takes place in the aqueous phase, i.e. organic solvents need not necessarily be purified off. According to the invention it is preferred that HAS binds directly to a ε—NH 2 -group, α—NH 2 -group, SH-group, COOH group or —C(NH 2 ) 2 -group of the active ingredient. Alternatively, a further reactive group can be introduced into HAS or the bond between HAS and the active ingredient can take place via a linker. The use of the corresponding linkers for the binding of active ingredients to PEG is known in the state of the art. The use of amino acids, in particular glycine, alanine, leucine, isoleucine, and phenylalanine, as well as hydrazine and oxylamine derivatives as linkers, as disclosed in WO 97/38727 and EP 605 963, the disclosures which are hereby incorporated by reference, is preferred. According to one embodiment of the process of the present invention, HAS is oxidized before binding to the active ingredient. The oxidation can take place according to one of the processes known in the state of the art, a selective oxidation of the reducing end groups of HAS being preferred. This facilitates processes in which the oxidized reducing end group of the HAS reacts with an amino group of the active ingredient resulting in the formation of an amide. This embodiment has the particular advantage that a specific bond between HAS and the active ingredients, and thus a particularly homogeneous product, is achieved. HAS can be reacted with oxidized reducing end groups and the active ingredient preferably for at least 12, most preferably at least 24 hours. Furthermore, it can be desirable to add any activator, for example ethyldimethyl-aminopropyl-carbodiimide (EDC). The molar ratio between HAS and the active ingredient during the reaction can be randomly selected, but is normally in the range of HAS:active ingredient of 20:1 to 1:20, a ratio of 6:1 to 1:6 being particularly preferred. The best results were achieved with a molar ratio of HAS:active ingredient of approx. 2:1. Other coupling reactions between an amino group of the active ingredient and HAS are naturally also comprised in the scope of the invention, for example, processes in which HAS and the active ingredient are reacted directly with each other, a Schiff's base forming between HAS and active ingredient as intermediate product. The azomethin group —CH═N— of the Schiff's base can then be reduced with formal addition of <2H> to a methyleneamine group —CH 2 —NH—. For this reduction, a person skilled in the art can use a number of reduction agents known in the state of the art, reduction using BH 4 is particularly preferred. The HAS can be coupled with any group of the active ingredient. The coupling is preferably carried out such that the conjugate displays at least 50%, preferably at least 75% of the activity of the active ingredient before coupling, a retention of at least 95% of the activity being particularly preferred. The coupling reaction can naturally also be controlled such that the HAS is bound exclusively to one or more specific groups of the active ingredient, for example to lysine or cysteine residues of a peptide. Particular advantages result if HAS is bound to one or more specific groups of active ingredient via the oxidized reducing end groups, as homogenous HAS-active ingredient conjugates are obtained with a corresponding process. According to a preferred embodiment of the process of the present invention, HAS and a protein or a peptide are used as starting substances for the reaction. Any therapeutic or diagnostic proteins of natural or recombinant origin can be used. A list of the active ingredients of recombinant preparation currently on the market is to be found in Pharma Business, July/August 2000, pages 45 to 60. The present invention comprises the preparation of HAS-active ingredient conjugates which comprise any one of the active ingredients named in this publication. The preparation of conjugates using cytokines, for example interferons, interleukins, growth factors, enzymes, enzyme inhibitors, receptors, receptor fragments, insulin, factor VIII, factor IX, antibiotics (or antiinfectives), peptide antibiotics, viral coat proteins, haemoglobins, erythropoietin, albumins, hTPA, antibodies, antibody fragments, single-chain antibodies, DNA, RNA or a derivative thereof is particularly preferred. Particular advantages result when using recombinant proteins or peptides in the process according to the invention. As already stated, corresponding proteins can often not be used as active ingredients due to their properties being antigenic for humans. After coupling to HAS by the processes according to the invention, however, the immunogenicity of the recombinant proteins decreases, which allows the medical use on humans. Furthermore, particular advantages result upon coupling of HAS to proteins having a short chain and smaller peptides. Currently, a large number of peptide libraries are being produced, for example, phage display libraries wherein short oligopeptides (for example 3 to 20 mers) are expressed on the surface of phages. Furthermore, antibodies from a single polypeptide chain (so-called “single chain antibodies”) are expressed in bacteria or on the surface of phages. These libraries are screened for specific active ingredient or binding activity. Hitherto, the therapeutic and diagnostic use of corresponding peptide active ingredients or antibodies has however failed because these are very quickly excreted from the organism due to their small size (e.g. Chapman et al., 1999, loc. cit.). With the process according to the invention, these peptides can advantageously be coupled to HAS and have an in vivo half-life which allows to use the same as an active ingredient. As an alternative to the above embodiment, a hormone, a steroid hormone, a hormone derived from amino acids or a hormone derived from fatty acids can be used as active ingredient. In the specific case, it may be necessary to introduce an active group into the hormone before binding to HAS, for example by using a linker. According to the invention, any physiologically compatible HES can be used as starting material. HES with an average molecular weight (weight average) of 1 to 300 kDA, in particular of 1 to 150 kDa is preferred. HES with an average molecular weight of 2 to 40 kD is particular preferred. HES preferably has a molar degree of substitution of 0.1 to 0.8 and a ratio of C 2 :C 6 substitution in the range of 2 to 20, in each case relative to the hydroxyethyl groups. The invention furthermore relates to the HAS-active ingredient conjugates obtainable according to the above processes. These conjugates have advantageous properties, namely high activity of the active ingredient, low immunogenicity, long residence time in the body and excellent Theological properties, which increase the medicinal benefit of the conjugates. Accordingly, the present invention likewise comprises processes for preparing a medicament or diagnostic in which a HAS-active ingredient conjugate is prepared according to one of the above processes and mixed with a pharmaceutically compatible carrier, adjuvant or auxiliary known in the state of the art, as well as medicaments or diagnostics obtainable according to this process. The medicinal use of the corresponding medicament depends on the type of active ingredient used. If, for example, haemoglobin is used as active ingredient, the conjugate can be used as an oxygen transport agent. If on the other hand, a cytokine is used as active ingredient for the preparation, the conjugate can for example be used in tumor therapy. The concentration of the conjugate to be used in each case in the medicament can be ascertained immediately in dose-effect tests by any average person skilled in the art. The diagnostics can be used in vivo or in vitro to diagnose illnesses or disorders. If an antibody or antibody fragment is used as active ingredient, the conjugate is suitable, for example, for carrying out the ELISA detection processes customary in the state of the art. In the examples, the materials described in the following were used: Human serum albumin: Sigma-Aldrich A3782 HES 130 kD: Type 130/0.5, prepared from T91SEP (Fresenius Kabi) Data: M w : 130 000 ± 20 000 D M n : 42 600 D HES 10 kD: Type HHH 4-2 (Fresenius Kabi) Data: M w : 9 800 D M n : 3 695 D EDC: Sigma-Aldrich no. 16.146-2 (ethyldimethyl aminopropyl carbodiimide) HOBt Sigma-Aldrich no. 15.726-0 (1-hydroxy-1H- benzotriazolhydrate) DIG glycan detection kit: Roche-Boehringer no. 1142 372 The following examples 1 to 6 describe the coupling of HSA and diaminobutane to HES with oxidized reducing end groups or the direct coupling of HSA to HES. HSA and diaminobutane are simply examples of the active ingredients defined above. Example 7 describes the coupling of oligonucleotides to HES. |
Arthropodicidal compositions |
Fluid compositions for the eradication or control of arthropods comprise a dispersion of at least one siloxane compound as an emulsion in a water continuous phase and having a dispersed particle size in the range 1 to 10 nm, preferably 10 to 50 nm. In a preferred embodiment the siloxane compound or compounds in the dispersed phase comprise from 50 to 99.9 percent by volume of one or more volatile siloxanes with the remainder one or more non-volatile siloxanes. The dispersed phase may have from 0.1 to 15 percent by volume of one or more non-volative siloxanes and from 97.1 to 95.5 percent by volume of one or more volatile siloxanes. The non-volative siloxane is preferably a linear siloxane such as dimethicone or dimethiconol and the volatile siloxane is a cyclic siloxane such as cyclopentasiloxane or cyclomethicone. Arthropods are controlled or eradicated by applying the fluid compositions to a surface which carries, or is believed to carry, arthropods. The water phase is lost leaving the siloxane compound or compounds. |
1. A fluid composition for the eradication or control of arthropods comprising a dispersion of at least one siloxane compound as an emulsion in a water continuous phase and having a dispersed particle size in the range 1 to 100 nm. 2. The fluid composition as claimed in claim 1 in which the dispersed particle size is in the range 10 to 50 nm. 3. The fluid composition as claimed in claim 1 or claim 2 in which the siloxane compound or compounds in the dispersed phase comprise from 50 to 99.9 percent by volume of one or more volatile siloxanes with the remainder one or more non-volatile siloxanes. 4. The fluid composition as claimed in claim 3 in which the siloxane compound or compounds in the dispersed phase comprise from 0.1 to 15 percent by volume of one or more non-volatile siloxanes. 5. The fluid composition as claimed in claim 3 in which the siloxane compound or compounds in the dispersed phase comprise from 97.1 to 95.5 percent by volume of one or more volatile siloxanes. 6. The fluid composition as claimed in any of the preceding claims 3 to 5 in which the non-volatile siloxane is a linear siloxane. 7. The fluid composition as claimed in claim 6 in which the non-volatile siloxane is dimethicone or dimethiconol. 8. The fluid composition as claimed in any of the claims 3 to 5 in which the volatile siloxane is a cyclic siloxane. 9. The fluid composition as claimed in claim 8 in which the volatile siloxane is cyclopentasiloxane or cyclomethicone. 10. The method of controlling arthropods comprising applying to a surface a fluid composition as claimed in any of the claims 1 to 9. |
Process and apparatus for friction stir welding |
The invention provides a method for a friction stir welding process, which comprises the steps of conventional stir welding—including applying friction to the areas of the workpiece to be welded by means of a rotating tool (20) that has a large shoulder that is pressed downwards on the workpiece and has a probe inserted into the material to be welded. The tool being advanced along the weld line. The method additionally comprises generating a laser beam (28) and collimating and focusing said beam on the workpiece in the weld region (29) ahead of the rotating tool. The invention also provides an apparatus for friction stir welding, which comprises the elements of conventional stir welding apparatus—including a rotating tool (20) that has a large shoulder and has a probe for insertion into the material to be welded, a mechanism for holding the tool and rotating and advancing it, a mechanism for pressing the tool shoulder downwards on the workpiece, and a mechanism for clamping the workpieces. The apparatus further comprises a laser beam generator (25), a laser beam conduit (26), preferably consisting of optical fibers, and collimator (28) and focusing optics for focusing the laser beam on the desired area of the workpiece. |
1. Friction stir welding process, which comprises the elements of conventional stir welding and additionally comprises generating a laser beam, leading, collimating and focusing said laser beam on the desired area of the workpiece. 2. Friction stir welding process according to claim 1, wherein the power applied by the laser beam is such as to bring the workpiece to a temperature that is comprised between 0.4Tm and Tm, where Tm is the melting temperature, in degrees Kelvin, of said workpiece. 3. Apparatus for friction stir welding, which comprises the elements of conventional stir welding apparatus—including a rotating tool that has a large shoulder and has a probe for insertion into the material to be welded, a mechanism for holding the tool and rotating and advancing it, a mechanism for pressing the tool shoulder downwards on the workpiece, and a mechanism for clamping the workpieces—and further comprises a laser beam generator, a laser beam conduit, preferably consisting of optical fibers, and collimator and focusing optics for focusing the laser beam on the desired area of the workpiece. 4. Apparatus for friction stir welding according to claim 3, wherein the power of the laser beam is such as to bring the workpiece to a temperature from 0.4Tm to Tm, where Tm is the melting temperature, in degrees Kelvin, of the workpiece. 5. Apparatus for friction stir welding according to claim 4, wherein the laser beam generator is a water cooled Nd:YAG laser. 6. Apparatus for friction stir welding according to claim 3, wherein a simple, relatively inexpensive machine comprises the mechanism for clamping the workpieces, the mechanism for holding the tool and rotating and advancing it, and the mechanism for pressing the tool shoulder downwards on the workpiece. 7. Apparatus for friction stir welding according to claim 6, wherein the simple, relatively inexpensive machine is a conventional milling machine. 8. Apparatus for friction stir welding according to claim 3, wherein the laser beam conduit consists of conventional laser beam steering optics. 9. Apparatus for friction stir welding according to claim 3, wherein the laser beam conduit consists of optical fibers. 10. Apparatus for friction stir welding according to claim 3, wherein the collimator comprises any type of collimating system based on reflective, diffracting, or refractive optics, preferably a single element collimating lens. 11. Apparatus for friction stir welding according to claim 3, wherein the focusing optics comprises any type of focusing system based on reflective, diffracting, or refractive optics, preferably a single element focusing lens. 12. Apparatus for friction stir welding according to claim 9, wherein no focusing means are employed for focusing the laser beam that exits from the optical fibers onto the desired area of the workpiece. 13. Apparatus for friction stir welding according to claim 3, wherein absorptive coatings are applied to the area of the weld in order to increase the absorption of laser energy. 14. Apparatus for friction stir welding according to claim 3, further comprising a mechanism for rotating and/or displacing the collimator and focusing optics and/or the optical fiber conduit and/or the laser generator, to keep the laser beam focused on the desired areas of the workpiece as the rotating tool progresses along the weld path. |
<SOH> BACKGROUND OF THE INVENTION <EOH>Friction stir welding is a welding process in which the parts to be joined are plasticized at and in the vicinity of their contact surfaces by heat generated by friction. Said parts to be joined will be called hereinafter, together, “the workpiece”. In the typical form of this process (see “Friction Stir—Where We Are and Where We are Going” by Wayne Thomas et al., TWI Bulletin, Vol. 39, May/June 1998), friction is generated between the workpiece and a rotating tool (of harder material than the workpiece), to plasticize the abutting weld region. Commonly, the tool is shaped with a large diameter shoulder and a specially profiled tool of small diameter (called hereinafter “the probe”) which is plunged into the joint region and is rotated while pressure is exerted on the said shoulder to force it downwards onto the workpiece. The friction stir welding process is disclosed in WO 93/10935. A modification of the process is described in WO 99/39861, and consists in applying a moving induction coil as a heat source in front of the rotating tool, to provide controlled heating of a limited volume of the weld material beneath the tool shoulder and plasticize it. In this way, it is said that the main function of the rotating probe is to control the flow pattern of the preheated material and to break up outside skin introduced from the welded members. The said modification of the friction stir welding process, however, is not fully satisfactory. All conductive materials, that are affected by the current passing through the induction coil, are heated, including clamping devices and even the tool probe, which is highly undesirable. Induced currents may flow across the path of the weld and cause spark formation. The heating by means of induction coil, of course, applies to conducting materials only and not, for example, to plastics or ceramics. Actually, the present technology of friction stir welding, in all its forms, is presently mainly applied to aluminum and aluminum alloys although it is suitable to other metals and also to non-metallic materials such as plastics and medium and low melting point ceramics. It is therefore a purpose of this invention to provide a friction stir welding process and apparatus that are free of the drawbacks of the known stir welding processes and apparatus. It is another purpose to provide a friction stir welding process and apparatus in which the parts to be joined are heated by means that are applicable to all materials. It is a further purpose to provide a friction stir welding apparatus and method in which heat is applied to the workpiece in a precise and localized manner such that both the location of heating and amount of power reaching the workpiece are known and predetermined. It is a still further purpose to provide a friction stir welding apparatus and method that permits successfully to weld high-melting temperature materials. Other purposes and advantages of the invention will appear as the description proceeds. |
<SOH> SUMMARY OF THE INVENTION <EOH>The process of the invention comprises the steps of conventional stir welding—including applying friction to the areas of the workpiece to be welded by means of a rotating tool that has a large shoulder that is pressed downwards on the workpiece and has a probe inserted into the material to be welded, said tool being advanced along the weld line—and additionally comprises generating a laser beam and collimating and focusing said beam on the workpiece in the weld region ahead of the rotating tool. The power applied by the laser beam depends on the nature of the material being welded, the dimensions of the workpiece, the characteristics of the rotating tool, and the heat dispersion from the workpiece, and therefore cannot be generally specified. However, the values of power that are required from the laser can be easily determined in each individual instance, since it must be such that it can raise the temperaure of the workpiece to a temperature that is comprised between 0.4T m and T m where T m , is the melting temperature of the workpiece, in degrees Kelvin, before the tool begins to rotate. Typical starting temperatures for different materials are: Mg≅300° C. Al≅300°-350° C. Cu≅600° C. Steel≅700° C. Ceramics≅2000° C. The laser beam may be generated by any laser system capable of producing enough power to achieve the desired temperature of the workpiece. The laser beam is led to the collimating and focusing elements by an optical fiber cable or by other optical means. The temperature of the workpiece that is heated by the laser beam is monitored on-line by any conventional temperature measuring device, for example a thermocouple or infrared temperature measuring camera. Preliminary calibration of the temperature of the workpiece as a function of laser energy and time of heating may alternatively be employed to infer its temperature. The invention also includes an apparatus for friction stir welding, which comprises the elements of conventional stir welding apparatus—including a rotating tool that has a large shoulder and has a probe for insertion into the material to be welded, a mechanism for rotating and advancing it and a mechanism for pressing the tool shoulder downwards on the workpiece. In the preferred embodiment of the invention, the mechanisms for holding, rotating and pressing the tool sholder down onto the workpiece as well as the mechanisms for clamping the workpieces and advancing them relative to the tool are all contained in a single machine. The invention further comprises a laser beam generator, a laser beam conduit, preferably consisting of optical fibers, and a collimator and focusing optics for focusing the laser beam on the desired area of the workpiece. It is also possible to use the optical fibre cable without any focusing optics to heat the workpiece. In other embodiments of the invention, conventional laser beam steering optics, well known to persons skilled in the art, are employed instead of the fibre optic cable to lead the beam to the desired area on the workpiece. The laser beam generator is chosen from among comercially available solid state, liquid, or gaseous lasers. The optical components preferably comprise a single element collimating lens and a single element focusing lens, but can be any type of collimating and focusing systems based on reflective, diffracting, or refractive optics. Further, the apparatus comprises a mechanism for rotating and/or displacing the collimator and focusing optics and/or the optical fiber conduit and/or the laser generator, to keep the laser beam focused on the desired areas of the workpiece as the rotating tool progresses along the weld path. While such mechanism can be manually actuated by an operator, it can be actuated by a controller as a function of the displacements of the rotating tool, and in this case the controller is a part of the apparatus of the invention. |
Pmi syrup phases and their use for stabilizing insoluble additives |
By using a high molecular weight syrup phase comprising PMMA or PMMI, it is possible to distribute fillers in the polymerization batch so that the distribution of the fillers remains homogeneous during the polymerization. A homogeneous foam results. |
1: A process for the preparation of a copolymer of a vinylically unsaturated carboxylic acid and a vinylically unsaturated carboxylic acid derivative by a casting method, wherein the polymerization is carried out in a highly viscous solution, a syrup phase, of a polymer soluble in a monomer mixture required for the preparation of polymethacrylimide. 2: A process for the preparation of a copolymer of a vinylically unsaturated carboxylic acid and a vinylically unsaturated carboxylic acid derivative by a casting method in the presence of a syrup phase, wherein the syrup phase is a high molecular weight polymethyl methacrylate or polymethacrylimide. 3: The process according to claim 1, wherein the vinylically unsaturated carboxylic acid is methacrylic acid. 4: The process according to claim 1, wherein the vinylically unsaturated carboxylic acid derivative is methacrylonitrile. 5: The process according to claim 1, wherein the weight ratio of the vinylically unsaturated carboxylic acid and of the vinylically unsaturated carboxylic acid derivative is between 80:20 and 40:60. 6: The process according to claim 1, wherein the amount of syrup phase is between 0.005 and 0.6 gram of polymer per gram of monomer mixture, the monomer mixture accounting for the sum of the masses of the vinylically unsaturated monomers. 7: The process according to claim 2, wherein the syrup phase comprises polymethyl methacrylate and the number average molecular weight of the polymethyl methacrylate is between 1 000 000 g/mol and 12 000 000 g/mol. 8: The process according to claim 2, wherein the syrup phase comprises polymethacrylimide and the number average molecular weight of the polymethacrylimide is between 50 000 g/mol and 500 000 g/mol. 9: A material prepared by the process according to claim 1, comprising between 0 and 400 parts by weight, based on the total mass of polymerizable groups, of fractions which are insoluble in the reaction mixture required for the preparation of the material. 10: A material prepared by the process according to claim 1, comprising between 0 and 400 parts by weight, based on the total mass of polymerizable groups, of fractions which are soluble in the reaction mixture required for the preparation of the material. 11: The material according to claim 9, wherein the material is a foam. |
<SOH> FIELD OF THE INVENTION <EOH>The invention relates to 1. PMI syrup phases through polymer additives 2. use of the abovementioned phases for stabilizing insoluble additives The PMI syrup phases are polymerized in a first step and foamed in a second step. The foam product is available under the brand Rohacell® from Rohm GmbH. Rohacell® is a closed-pore foam, which is used, for example, in automotive construction and in aircraft construction. |
Novel macrolide derivatives havaing effect of potentiating antifungal activity |
Macrolide derivatives having enhancing effect for activities of azole antifungal agents, acting at low concentration and within a short time against fungal infection and reducing the frequency of appearance of resistant microorganisms. One such substance is a compound represented by the formula [I]: wherein R1 is Ac, R2 and R3 are Ac, and R4 is Me; when R1 is H, R2 and R3 are Ac, and R4 is Me; when R1 is H, R2 and R3 are Ac, and R4 is H, when R1 Bzl, R2 and R3 are Bzl, and R4 is Me; when R1 is Ac, R2 and R3 are Pr, and R4 is Me; when R1 is Ac, R2 and R3 are Hex, and R4 is Me; when R1 is Ac, R2 and R3 are Bzl, and R4 is Me; when R1 is H, R2 and R3 are Pr, and R4 is Me; when R1 is H, R2 and R3 are Hex, and R4 is Me; when R1 is H, R2 and R3 are Bzl, and R4 is Me; when R1 is H, R2 is H, R3 is Bzl, and R4 is Me; when R1 is H, R2 and R3 are Hex, and R4 is H, or when R1 is H, R2 and R3 are Hex, and R4 is Et. |
1. Novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [I]: wherein when R1 is Ac, R2 and R3 are Ac, and R4 is Me; when R1 is H, R2 and R3 are Ac, and R4 is Me; when R1 is H, R2 and R3 are Ac, and R4 is H; when R1 is Bzl, R2 and R3 are Bzl, and R4 is Me; when R1 is Ac, R2 and R3 are Pr, and R4 is Me; when R1 is Ac, R2 and R3 are Hex, and R4 is Me; when R1 is Ac, R2 and R3 are Bzl, and R4 is Me; when R1 is H, R2 and R3 are Pr, and R4 is Me; when R1 is H, R2 and R3 are Hex, and R4 is Me; when R1 is H, R2 and R3 are Bzl, and R4 is Me; when R1 is H, R2 is H and R3 are Bzl, and R4 is Me; when R1 is H, R2 and R3 are Hex, and R4 is H; or when R1 is H, R2 and R3 are Hex, and R4 is Et. 2. Novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [II]: wherein when R1 is Ac, R2 is SO2Ph; when R1 is Ac, R2 is SO2Bn, or when R1 is H, R2 is SO2Bn. 3. Novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [III]: 4. Novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [IV]: wherein R1 is H or Me. 5. Novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [V]: wherein R1 is H or Me. 6. A novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [VI]: 7. A novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [VII]: 8. A novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [VIII]: 9. A novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [IX]: 10. A novel macrolide derivative having enhancing effect for activities antifungal agent represented by the formula [X]: 11. A novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [XI]: 12-36. (canceled) 37. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 1. 38. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 2. 39. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 3. 40. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 4. 41. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 5. 42. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 6. 43. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 7. 44. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 8. 45. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 9. 46. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 10. 47. A method for enhancing the activities of antifungal agents for the prevention or treatment of fungal infections accompanied by immunocommpromised conditions caused by HIV infection or blood diseases, comprising administering to a person in need of the same therapeutically effective amounts of a said antifungal agent and a derivative as claimed in claim 11. |
<SOH> BACKGROUND OF THE INVENTION <EOH>The present invention relates to a substance enhancing an effect of antifungal agent used for fungal infection. More particularly, the present invention relates to novel macrolide derivatives having enhancement action for antifungal activity which can enhance antifungal effect in combination with azole antifungal agent used in chemmotherapy for fungal infection accompanied by immunocormmpromised condition such as HIV infection and blood disease. |
<SOH> SUMMARY AND OBJECTS OF THE INVENTION <EOH>In diseases accompanied with immunocommpromised condition such as HIV infection and blood disease, compromised condition is generated as a result, incidence of fungal infection as opportunistic infection is increased. Many cases of diseases accompanied with immunocommpromised condition are severe and required long-term therapy. For that reason, chemotherapy of fungal infection requires for long term, and drug resistance is easily induced in frequently used azole antifungal agents. Proposed mechanisms of resistance for azole antifungal agents are: for example, excess expression of P-450 14-α-demethylase, a target enzyme, in Candida albicans , and reduced affinity with drugs due to amino acidmutation (VandenBossche, H. et al. Antimicrob. Agents and Chemoth., 36, 2602-2610, 1992; Sanglard, D. et al. ibid. 42, 241-253, 1998); reduced intracellular drug concentration by an action of multiple drug excretion transporter such as MSF (major facilitator superfamily) and ABC (ATP binding cassette) (Fling, M. E. et al Molecular Genetics and Genomics, 227, 318-329, 1991; Sanglard, D. et al., Microbiology, 143, 405-416, 1997). MDR (multiple drug resistant) genes, PDR 16 and PDR 17, are involved in the lipid metabolism in Saccharomyces cerevisiae , and in case of deficient in these genes, microorganisms can be high susceptive against azole compounds (H. Bart van den Hazel et al. J. Biol. Chem., 274, 1934-1941, 1999). Consequently, drugs which can increase activity of azole antifungal agents can be expected to decrease dosage of drugs and to shorten administration term, as a result, frequency of generation of resistant microorganisms can be decreased. At the same time, combined use of two types of drugs having different skeletal structures or combined use of such drugs for resistant strains against azole compounds are expected to overcome resistance against azole antifungalagents. Consequently, providing drugs having enhanced action for activity of azole antifungal agents is thought to be useful for prevention and therapy of fungal infection and azole resistant fungal infection caused by such as deep sheeted mycosis. In such conditions, we have studied extensively an action of various macrolide derivatives for enhancing activities of azole antifungal agents, and found the action for enhancing activities of azole antifungal agents in the macrolide derivatives having novel skeletal structure which is different from known azole antifungal agents used for treatment of fungal diseases, and have completed the present invention. An object of the present invention is to provide novel macrolide derivatives having enhancing effect for activities of antifungal agents, acting at low concentration and within short term against fungal infection and making possibility for reducing frequency of appearence of resistant microorganisms. The present invention also provides novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [I]: wherein when R1 is Ac, R2 and R3 are Ac, respectively, and R4 is Me; when R1 is H, R2 and R3 are Ac, respectively, and R4 is Me; when R1 is H, R2 and R3 are Ac, respectively, and R4 is H; when R1 is Bzl, R2 and R3 are Bzl, respectively, and R4 is Me; when R1 is Ac, R2 and R3 are Pr, respectively, and R4 is Me; when R1 is Ac, R2 and R3 are Hex, respectively, and R4 is Me; when R1 is Ac, R2 and R3 are Bzl, respectively, and R4 is Me; when R1 is H, R2 and R3 are Pr, respectively, and R4 is Me; when R1 is H, R2 and R3 are Hex, respectively, and R4 is Me; when R1 is H, R2 and R3 are Bzl, respectively, and R4 is Me; when R1 is H, R2 is H and R3 are Bzl, and R4 is Me; when R1 is H, R2 and R3 are Hex, respectively, and R4 is H; or when R1 is H, R2 and R3 are Hex, respectively, and R4 is Et. The compound represented by the above formula [I] has an action to enhance activities of azole antifungal agents against Candida albicans and Aspergillus niger , both of which include resistant strains. The present invention provides novel macrolide derivatives having enhancing action of antifungal agent against, for example, Candida albicans such as Candida albicans ATCC 64548 or Candida albicans ATCC 64550, and Aspergillus niger such as Aspergillus niger ATCC 6275. The present invention also provides novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [II]: wherein when R1 is Ac, R2 is SO2Ph; when R1 is Ac, R2 is SO2Bn; or when R1 is H, R2 is SO2Bn. The compound represented by the above formula [II] has an action to enhance activities of azole antifungal agents against Candida albicans and Aspergillus niger , both of which include resistant strains. The present invention provides novel macrolide derivatives having enhancing action of antifungal agent against, for example, Candida albicans such as Candida albicans ATCC 64550, and Aspergillus niger such as Aspergillus niger ATCC 6275. The present invention also provides novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [III]: The compound represented by the above formula [III] has an action to enhance activities of azole antifungal agents against Candida albicans and Aspergillus niger , both of which include resistant strains. The present invention provides novel macrolide derivatives having enhancing action of antifungal agent against, for example, Candida albicans such as Candida albicans ATCC 6458, and Aspergillus niger such as Aspergillus niger ATCC 6275. The present invention also provides novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [IV]: wherein R1 is H or Me. The compound represented by the above formula [IV] has an action to enhance activities of azole antifungal agents against Candida albicans and Aspergillus niger , both of which include resistant strains. The present invention provides novel macrolide derivatives having enhancing action of antifungal agent against, for example, Candida albicans such as Candida albicans ATCC 64548, and Aspergillus niger such as Aspergillus niger ATCC 6275. The present invention also provides novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [V]: wherein R1 is H or Me. The compound represented by the above formula [V] has an action to enhance activities of azole antifungal agents against Candida albicans and Aspergillus niger , both of which include resistant strains. The present invention provides novel macrolide derivatives having enhancing action of antifungal agent against, for example, Candida albicans such as Candida albicans ATCC 64548, and Aspergillus niger such as Aspergillus niger ATCC 6275. The present invention also provides novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [VI]: The compound represented by the above formula [VI] has an action to enhance activities of azole antifungal agents against Candida albicans and Aspergillus niger both of which include resistant strains. The present invention provides novel macrolide derivatives having enhancing action of antifungal agents against, for example, Candida albicans such as Candida albicans ATCC 64548, and Aspergillus niger such as Aspergillus niger ATCC 6275. The present invention also provides novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [VII]: The compound represented by the above formula [VII] has an action to enhance activities of azole antifungal agents against Candida albicans and Aspergillus niger , both of which include resistant strains. The present invention provides novel macrolide derivatives having enhancing action of antifungal agent against, for example, Candida albicans such as Candida albicans ATCC 64548, and Aspergillus niger such as Aspergillus niger ATCC 6275. The present invention also provides novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [VIII]: The compound represented by the above formula [VIII] has an action to enhance activities of azole antifungal agents against Candida albicans . The present invention provides novel macrolide derivatives having enhancing action of antifungal agents against, for example, Candida albicans such as Candida albicans ATCC 64548. The present invention also provides novel macrolide derivative having enhancing effect for activities of antifungal agents represented by the formula [IX]: The compound represented by the above formula [IX] has an action to enhance activities of azole antifungal agents against Candida albicans . The present invention provides novel macrolide derivatives having enhancing action of antifungal agents against, for example, Candida albicans such as Candida albicans ARCC 64548. The present invention also provides novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by the formula [X]: The compound represented by the above formula [X] has an action to enhance activities of azole antifungal agents against Candida albicans and Aspergillus niger , both of which include resistant strains. The present invention provides novel macrolide derivatives having enhancing action of antifungal agent against, for example, Candida albicans such as Candida albicans ATCC 64548, and Aspergillus niger such as Aspergillus niger ATCC 6275. The present invention also provides novel macrolide derivatives having enhancing effect for activities of antifungal agents represented by-the formula [XI]: The compound represented by the above formula [XI] has an action to enhance activities of azole antifungal agents against Candida albicans . The present invention provides novel macrolide derivatives having enhancing action of antifungal agent against, for example, Candida albicans such as Candida albicans ATCC 64548. The present invention provides use of any one of compounds selected from the group consisting of compounds represented by the formula [I] to [XI] for production of pharmaceuticals for enhancing activities of antifungal agents for prevention or treatment of fungal infection accompanied by immunocommpromised condition caused by HIV infection or blood disease. The present invention also provides a substance of any one of compounds selected from the group consisting of compounds represented by the formula [I] to [XI] for production of pharmaceuticals for enhancing activities of antifungal agents for prevention or treatment of fungal infection accompanied by immunocommpromised condition caused by HIV infection or blood diseases. Compound No. of macrolide derivatives represented by the formula [I] of the present invention, R1, R2, R3 and R4 in the formula, and numbers of examples are listed as follows. Compound No. R1 R2 R3 R4 Example No. EM719 Ac Ac Ac Me 1 EM755 H Ac Ac Me 2 EM756 H Ac Ac H 3 EM770 Bzl Bzl Bzl Me 4 EM771 Ac Pr Pr Me 5 EM772 Ac Hex Hex Me 10 EM773 Ac Bzl Bzl Me 6 EM776 H Pr Pr Me 7 EM777 H Hex Hex Me 11 EM778 H Bzl Bzl Me 8 EM779 H H Bzl Me 9 EM852 H Hex Hex H 12 EM853 H Hex Hex Et 13 Compound No. of macrolide derivatives represented by the formula [II] of the present invention, R1 and R2 in the formula, and numbers of examples are listed as follows. Compound No. R1 R2 Example No. EM774 Ac SO2Ph 14 EM775 Ac SO2Bn 15 EM780 H SO2Bn 16 Compound No. of macrolide derivatives represented by the formula [III] of the present invention, R1 in the formula, and numbers of examples are listed as follows. Compound No. R1 Example No. EM762 17 EM763 18 EM769 —CH 2 CH 2 CH = CH 2 19 Compound No. of macrolide derivatives represented by the formula [IV] of the present invention, R1 in the formula, and numbers of examples are listed as follows. Compound No. R1 Example No. EM752 H 20 EM753 Me 21 Compound No. of macrolide derivatives represented by the formula [V] of the present invention, R1 in the formula, and numbers of examples are listed as follows. Compound No. R1 Example No. EM757 H 22 EM758 Me 23 Compound No. of macrolide derivatives represented by the formula [VI] of the present invention, and numbers of examples are listed as follows. Compound No. R1 Example No. EM759 24 Compound No. of macrolide derivatives represented by the formula [VII] of the present invention, and numbers of examples are listed as follows. Compound No. Example No. EM760 25 Compound No. of macrolide derivatives represented by the formula [VIII] of the present invention, and numbers of examples are listed as follows. Compound No. Example No. EM761 26 Compound No. of macrolide derivatives represented by the formula [IX] of the present invention, and numbers of examples are listed as follows. Compound No. Example No. EM764 27 Compound No. of macrolide derivatives represented by the formula [X] of the present invention, and numbers of examples are listed as follows. Compound No. Example No. EM765 28 Compound No. of macrolide derivatives represented by the formula [XI] of the present invention, and numbers of examples are listed as follows. Compound No. Example No. EM741 29 detailed-description description="Detailed Description" end="lead"? |
Method for identification by a host computer avoiding collision of elements |
An anti-collision process for elements controlled by a host computer allowing the identification of elements by the latter, each of the elements having an identification number between 0 and a maximum value (MAX), and including transmission by the host computer of a query instruction able to be received and recognized by all elements, transmission by the host computer of an anti-collision instruction including an identification attempt digital value, response by any element having an identification number less than or equal to the digital identification attempt value, and detection of a collision between several elements when the host computer receives several responses and transmissions of a new anti-collision instruction according to a given algorithm, or selection of the element by its identification number when there is only one response. |
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