JPH07500843A - How to suppress restenosis - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] How to suppress restenosis field of invention The present invention relates to compounds, compositions and methods for healing wounded living tissue. In particular, compounds with a basic structure of saccharides that remain at the wound site for a long time and and compositions.

Background of the invention When a tissue experiences injury, a series of events is initiated that result in tissue repair and wound healing. It will be done. During the first few days after injury, neutrophils, macrophages, and and fibroblasts migrate. Macrophages and fibroblasts that migrate to the wound site activated, thereby promoting endogenous growth factor production and provisional extracellular matrix synthesis. formation, fibroblast proliferation and collagen synthesis. Eventually, the wound was sustained. Approximately 2 weeks to 1 year later, the wound is folded back and reorganized and cross-linked using activated collagen. (Pierce et al., 1991. J.

Ce1l Biochem, 45:319-326). This repair process Cell proliferation, migration and protein synthesis are regulated, although little is known about the manner in which they are regulated. , which can be stimulated by growth factors that act on cells with their receptors. is known.

In vivo studies have shown that exogenous monotherapy is induced after experimental wounding in animals. - Local application of growth factors or combinations of growth factors accelerates the healing process. It was shown that (Antoniades et al., 199), P roc, Natl, Acad, Sci, IJSA, 8B:565-569). child It has been suggested that these growth factors have the ability to promote wound healing. Efforts are being made to have a child. Heparin-binding growth factor (HBGF) Some of these growth factors, known as heparin, have a strong affinity for heparin. is known (LObb et al., 1988. Fur, J., Cl. 1n, Invest.

18:321-328 and Folkman and Jellyfish Pla. (Klagsbrun, 1987.5science 235:442-447 ) Therefore, in order to obtain these growth factors in purified form, heparin affinity Tychromatography is used. In addition, some of these growth factors The encoding DNA is isolated and the protein is produced using recombinant DNA methods. Ru. These HBGFs are found in almost all mesoderm and neuroectoderm cells in vitro. It has been shown to have mitogenic and non-mitogenic effects on Yamaki cells. HBG F is also known to promote migration, proliferation and differentiation of these cells in vivo. It is. Therefore, Robb showed that HBGF leads to soft tissue repair. Instigated (1988. hr, J, Cl1n, Invest, 18:3 21-328) o Furthermore, HBGF can be used to repair hard tissues such as bones and cartilage. It was also suggested that it could be used. In contrast to their beneficial effects, growth Factors overstimulate the wound healing response, resulting in excessive smooth muscle cell proliferation and migration , these are also known to cause restenosis, for example after angioplasty.

The affinity of growth factors for heparin and the ability to obtain heparin in pure and homogeneous form. Although heparin has an affinity for growth factors, Attempts have been made to obtain compounds that can be easily and reproducibly produced. There is. One of the compounds meeting these requirements, as described in the parent application referenced above. The group consists of cyclodexts consisting of at least 6 glucobylanose units. Phosphorus, a cyclic oligosaccharide.

U.S. Patent No. 5019.562 to Volkmann et al. (Volkmann et al. patent) for treating unwanted cell or tissue growth. It is directed to the use of highly soluble cyclodextrin derivatives for this purpose.

The cyclodextrin derivatives disclosed in this patent combine with growth-inhibiting steroids. Administered together or alone, they absorb growth factors present in the bloodstream. Fol The cyclodextrin derivatives disclosed in the Kumann et al. patent are highly hydrophilic. Therefore, it is highly soluble. The high solubility of these derivatives makes them resistant to exogenous steroids. An important compound that interacts cooperatively with the innate conjugation ability of the cyclodextrin structure It is said that this is a factor. In addition, the high solubility of these compounds makes it difficult for them to be incorporated into the body. It is said to facilitate the introduction of compounds and aid in their dispersion through the bloodstream.

Summary of the invention The high solubility of the compounds disclosed in Volkmann et al.'s patent makes these compositions difficult to administer to the body. Desirable for systemic administration. However, on the other hand, the high solubility of these compounds Applicants have shown that limiting their ability to remain present in the wound area after administration I found it. Maximize the delivery of the administered growth factor or growth factors to the wound site. Satsuka, which has a high affinity for growth factors and very low solubility, can be used to Applicants have discovered that it is desirable to obtain compounds having a ride-based structure. I saw it. According to one aspect of Applicants' discovery, it is possible to introduce such low solubility compounds into the body. It is mixed with growth factors and applied topically to the wound site before administration. at least partially Due to their low solubility, such compounds remain at the site of application and grow slowly. release factors and optimize the dosage of growth factors at the wound site. Also, this application They found that compounds with both high affinity and low solubility for growth factors are retaining at least a portion of the growth factor at the site of injury and absorption into the injured tissue; can be used to reduce the risk of restenosis, as seen in post-angioplasty restenosis. It has been found that the possibility of over-stimulation of the wound healing process can be reduced.

In view of both the beneficial and aberrant properties of growth factors involved in wound repair, Applicants , concentration of growth factors and/or in the wound area to optimize the wound healing process. The need for a composition that modulates diffusion was identified. Therefore, the present invention provides living organisms of mammals. Low solubility polyanionic with high negative charge density to influence tissue growth A saccharide derivative is provided. In addition, low solubility polyanionic saccharide inducers an active material containing a physiologically acceptable carrier for the conductor and the saccharide derivative; Also provided are compositions containing sexual substances.

The saccharide derivative preferably has a body temperature soluble content of less than about 15 g per 100 ml of water. It has decomposability. According to certain preferred embodiments, the saccharide derivative has a temperature It has virtually no solubility in water. The term “body temperature” used here is The range of body temperatures expected in living mammals and used in various surgical procedures. the lower body temperature encountered during the physiological response to infection and the increased body temperature encountered in the physiological response to infection. include. Unless otherwise specified, solubility refers to solubility in distilled water.

Compositions of the invention are at least partially characterized by low solubility of the active ingredient in body tissues and fluids. The dissolvability provides many advantages over prior art compositions. Honsatsukaride induction The low solubility of the body is beneficial in wound healing methods that are administered directly to the wound site. Applicable The composition remains substantially at the site of administration for an extended period of time. When mixed with growth factors, the main Ming's saccharide derivatives promote the controlled release of growth factors at the wound site. , thereby regulating and greatly promoting the wound healing process. Without growth factors, Due to their affinity for growth factors, the present compositions are highly effective in stimulating cells at the wound site. local concentration and/or spread of growth factors produced in the bloodstream and present in the bloodstream. can reduce the dispersion. By reducing the diffusion of growth factors, the group The composition can prevent or substantially reduce overstimulation of the wound healing response; Resenosis after angioplasty, vein graft intimal hyperplasia and native vascular atherosclerosis The abnormal growth of cells that leads to symptoms such as tumor sclerosis can be avoided.

The present invention also provides methods of making beneficial wound healing compositions. These compositions are compared Contains a highly anionic polysaccharide in a physically insoluble, solid form. Method aspects reacts a saccharide with an anionic derivatizing agent to form a polyamine of the saccharide. After the formation of the ionic derivative, it is possible to form a salt, which is a highly insoluble product. including. Separately, saccharide can be reacted with a suitable coupling agent to reduce solubility. After forming a new polymer or copolymer of the saccharide, the anionic derivative react with a chemical agent. According to certain embodiments, these derived saccharides are then is mixed with one or more growth factors. Compositions provided by these manufacturing methods It has the advantageous properties of very low solubility and high growth factor affinity.

The invention also provides a method of wound healing. According to these methods, this low solubility polyanidine Apply the therapeutic saccharide to the area to be treated.

Such methods can prevent restenosis, promote angiogenesis, and treat transplanted tissues or organs. , and for use in the treatment of injured or transplanted bone or cartilage.

The ability of the present compounds and compositions to modulate the wound healing process may be useful in percutaneous transluminal angioplasty. (percutaneous lransluminal angioplas ty) (hereinafter referred to as “PCTA”) to save as much life as possible. do. Up to 40% of patients undergoing PCTA develop restenosis and recurrent arterial Suffering from recurrent blockage It is recognized that Thus, treatments for arteriosclerosis such as angioplasty Long-term efficacy is substantially limited by recurrence of restenosis. This composition prevents restenosis. substantially reduce or eliminate angioplasty, vein graft bypass surgery and likely to have a significant impact on morbidity and mortality in patients undergoing similar procedures . In addition, patients with cardiac, cerebral or peripheral ischemic disease may benefit from using the compositions of the present invention. It is thought that there will be great benefits from this. In particular, patients with infarcted myocardial tissue , requires the establishment of new collateral capillaries and new blood vessels that supply blood to the infarcted tissue. It is essential.

The composition merely promotes angiogenesis at the site of infarcted tissue.

Brief description of the drawing Figure 1 (A and B) shows (A) α-1β- and γ-cyclodextrin monomers. and (B) the two-dimensional shape of these cyclodextrin monomers. It is a depiction.

Figure 2 schematically shows the chemical structure of sucrose with the anionic substituent sites shown. It is depicted in.

Figure 3 shows tetradeca sulfate β-cyclodextrin against basic fibroblast growth factor. Shows the affinity of phosphorus polymers.

Figure 4 shows cyclodextrin copper viaaffinity chromatography. Matographically purified basic fibroblast growth factor and chondrosarcoma-derived growth Figure 3 shows polyacrylamide gel electrophoresis of factors. Lane l is protein marker - (phosphorylase b1 bovine serum albumin, ovalbumin, carbonic anhydrase, large Bean trypsin inhibitor, β-lactoglobulin, and lysozyme) Shows protein quality profile. Lanes 2 and 3 represent basic fibroblast growth, respectively. 18.000 molecular weight polypeptide band of factor and chondrosarcoma-derived growth factor is shown. .

Figure 5 shows heparin and tetradeca sulfate β-cyclodehyde against chondrosarcoma-derived growth factor. The present invention compares the affinity of kistrin polymers, and the present invention compares the affinity of kistrin polymers. The present invention is directed to compounds, compositions, and methods that affect hair growth. Novelty of the present invention The compound has low solubility in distilled water at body temperature and has a high negative charge density. It is a cyclodextrin polymer. The composition has a relatively high density of anionic Low solubility polyanionic saccharide derivatives having substituents and such derivatives Contains a carrier for.

One important aspect of the present compounds and compositions is that such It is a strong affinity for substances. Applicants do not necessarily wish to be bound or limited to any particular theory. Although it is not intended that the density of anionic groups on the saccharide compounds of the invention is important in conferring the high affinity of these compounds for tissues and growth factors. It is thought that Applicants have proposed that, in combination with the low solubility of the present saccharide derivatives, The affinity of the present compounds and compositions for growth factors in the wound area have been found to provide the ability to regulate and control the concentration of In addition, this compound and the composition contains active substances in the form of the present derivatized saccharides that tend to adhere to active tissues. Providing sexual substances. As a result, such compositions and compounds can be applied to the site of injury. It has the highly desirable ability to provide active wound healing agents for long periods of time.

The present invention provides methods for producing these compositions and various It is also directed to methods of treating wounds. As used herein, the term “wound healing” The term refers to the repair or reconstruction of cellular tissue. Wounds are the result of injuries or accidents such as burns. fruit may arise.

Wounds treatable by the present compositions and methods include all types of surgical procedures, i.e. catheterization or angioplasty that results in a wound to the surface of a blood vessel or organ. From a few invasive procedures to numerous surgical procedures such as bypass or organ transplant surgery. This includes surgical procedures. This concept of wound healing includes injured or fractured This includes bone or cartilage repair and promoting the establishment of bone grafts or grafts.

■6 compositions Applicants have proposed that polyanidines with high negative charge density and low solubility as active substances Compositions containing inflammatory saccharide derivatives may be useful as wound healing agents. I discovered that. Particularly preferred are polyanionic cyclodextrin polymers. be.

As used herein, the term “polyanionic saccharide derivative” broadly refers to , saccharides having 1.3 or more anionic substituents per sugar unit A compound whose basic structure is

The term sugar unit as used herein includes, for example, a hexose or a pentose. It refers to the basic monosaccharide building block.

Basic monosaccharides that serve as examples are glucose, fructose, amylo This is the case. All compounds containing the basic skeletal structure, as well as such compounds Homologs, analogs and isomers of are within the scope of "saccharide" as used herein. I believe that. The saccharide compounds mentioned here include, for example, disaccharide , Trisaccharide, Tetrasaccharide, Oligosaccharide, Polysacchara and polymers of such saccharides.

The term "oligosaccharide" means from about 650 to about 1 if unsubstituted Refers to a saccharide of about 5 to about IO sugar units having a molecular weight of 300. “Polysa The term ``caralide'' refers to sugar units containing more than the molecular weight or approximately IO. It means id. Polysaccharides have many different structures and substituents. It is understood to be a saccharide having sugar units. The polymer used here is The term refers to repeating structures or repeating structures based on monomers that are linked together to form a polymer. It refers to saccharide compounds similar to it.

Applicants have shown that the relationship between the structure of induced saccharides and the level of negative charge density is It has been discovered that the effectiveness of compounds, compositions and methods can be influenced. For example, The ionic substituents preferably have an average of about 1.0 to about 4 substitutions per sugar unit in the molecule. Exist within a range of groups. Particularly preferred compounds have an average of at least about It is a compound whose basic structure is a saccharide having a 1°4 anionic substituent.

For saccharide compounds composed of n sugar units and R substituents, the derivatized The anionic substituents on tucharide correspond substantially to approximately the following numerical values.

When n=2-3: Average anionicity R≧3.5 per n unit, when n=4-5 ; Average anionicity per n unit When R≧2.0, n≧6; Average anionicity per n unit Anionic R≧1.4゜The anionic substituent of the present invention includes known and introduceable anionic substituents. Anionic substituents can be chosen from a large group of anionic substituents; From ruboxyl, phosphoric acid, sulfonic acid groups, and combinations of two or more of these Generally preferred is selected from the group consisting of: Preferred compositions have 6 or more A saccharide having a sugar unit and having about 2 to about 3 substituents per sugar unit. and a saccharide in which the substituents include sulfuric acid, sulfonic acid and/or phosphoric acid substituents. Basic structure.

The saccharide derivatives of the present invention have low solubility in distilled water at body temperature. stomach. As used herein, the term "low solubility" refers to approximately 15 g/100 ml of water. The solubility is much lower than that of

It remains in a solid state for a considerable period of time in an aqueous medium such as physiological water or distilled water. related to the ability of the present saccharide compounds to

According to certain preferred embodiments, the saccharide derivative is dissolved in distilled water at body temperature. It has virtually no solubility in. That is, the solubility of the saccharide derivative is Much lower than about 1g per 100ml, more preferably about 1g per 100ml It can even be less than 1 mg. Such insolubility can be caused by, for example, polymer agglomerates or Using a saccharide composition comprising a dispersion of polymer particles that is physically solid Therefore, it is obtained. Although it is contemplated that a variety of particle sizes and shapes may be used, The particles have an average particle size of about i millimicrons to about 1000 microns in diameter. preferable. Expressed in terms of molecular weight, multiple polymers including this polymer have an average and has a molecular weight of about 100 million or more. This high molecular weight of the preferred polymer is , due to the presence of several hundred individual undissolved sugar units. In addition, the main In another embodiment of the present invention, the particles having the desired insolubility are polycationic moieties. form salts containing anionic saccharides in combination with or association with Manufactured by

Although the compositions of the present invention may be prepared from soluble saccharides as starting materials, As shown in Figure 2, most starting materials with poor solubility, such as cellulose or starch, can be used as starting materials. It is also possible to use solid or solid saccharides. These satsuka rides If a source is used, the solid saccharide is preferably broken down chemically or enzymatically. and then introducing substituents according to the invention.

A, cyclodextrin derivative Particularly preferred compositions according to the invention are compositions containing cyclodextrin derivatives. It is a thing. Cyclodextrins form cyclic or toroidal molecules. is also a saccharide compound containing 6 glucobylanose units and therefore It has no end groups.

Cyclodextrins with up to 12 glucobylanose units are known However, only the first three congeners have been extensively studied. These compounds are easily It has a simple, well-defined chemical structure as shown in Figure 1(A). Low molecular type The common names α-1β- and γ-cyclodextrin are used throughout this specification. Fig. 1, which is used in various cases, with n = 6.7 or 8 glucobylanose units, respectively. Corresponds to the chemical structure shown in (A). Cyclodex as a starch degradation product The first discovery of trines was made around the beginning of this century, and Schaldinger The compound is prepared by the action of Bacillus macerans amylase on starch. It was shown that it can be manufactured.

In older literature, the compound is often referred to as Schardinger dextrin. . They are also sometimes called cycloamyloses.

Topographically, cyclodextrin is located at its upper edge, as shown in Figure 1 (B). It is represented as a torus with primary -CH20H groups lined up and secondary hydroxyl groups lined up at the lower edge. can be done. Coaxially parallel to the torus are α-1β- and γ-shields. Channel-like caps of approximately 5.6 or 7.5 A, U, diameter for each clodextrin. It's Bitty. These cavities contain hydrophobic cyclodextrins of suitable diameter. It allows the formation of clathrate compounds with guest molecules.

The composition of the invention preferably comprises a polyanionic cyclodextrin derivative. . Generally, “induced CD”, “CD derivative” and similar terms refer to α(l→4) Bonded to carbon 2.3 or 6 of the CD molecule without breaking the hemiacetal bond Refers to chemically modified CDs formed by reaction of primary or secondary hydroxyl groups.

Details of such preparation methods are described in “Tetr. ahedron Report Number 147.5ynthesis of Chemically Modified Cyclodextrins” , A.P., Craft and Bartsch, Tet. rahedron 39(9):1417-1474 (1983) (hereinafter “ Tetrahedron Report No. 147″) There is.

The CD derivatives are preferably derived from derivatized cyclodextrin monomers, dimers, triesters, etc. polymer, or a mixture thereof. Generally, the cyclodextrin of the invention The derivative has at least 6 glucosols with an α(1→4) hemisecure linkage. Consisting of or formed from derivatized cyclodextrin monomer units consisting of villanose units be done. Preferred derivatized cyclodextrin monomers of the invention have the general formula: - at least two of said R groups per unit are anionic substituents, and the remaining said R groups are anionic substituents; If the R group is present, it is a non-alternative selected from well-known and introduceable substituents. It is a ionic group.

The remaining non-anionic R groups are, for example, ■], alkyl, aryl, ester, Ethers, thioesters, thioethers and -COOH may also be used. as an example Suitable alkyl groups include methyl, ethyl, propyl and butyl. Applicable The remaining non-anionic R groups may be hydrophilic or hydrophobic, depending on the particular needs of the desired composition. or a combination thereof. However, the solubility of the compound The remaining non-anionic R substituents are generally hydrophobic in order to minimize preferred.

For CD monomers having the structure 2, where n is about 6 to about 8, the compound An average of at least about 9 anionic R substituents per monomer unit, more preferably has at least about 12 anionic R substituents per monomer, even more preferably It is preferred to have at least about 14 anionic R substituents per monomer.

Generally, the anionic substituents are relatively uniformly distributed on the monomer molecule, so that n Compounds having a structure of formula I that is from about 6 to about 8 preferably have from about 1 to about 8 per n unit. about 3 anionic R substituents, more preferably about 1.3 to about 2.5 per n unit Anionic R substituents, even more preferably about 1.4 to about 2.2 a-units per n unit. It is preferred to have an ionic R substituent. Such structures have been shown to be therapeutically beneficial. and the highest charge found to provide high negative charge density and provide excellent results. It is believed to provide density molecules.

Polyanionic cyclodextrin monomers of the type described above are preferred according to the invention. It is an important component of the composition. The monomer unit may be, for example, an insoluble polymer or copolymer. in the form of polymeric structures or derivatized cyclodextrin monomers, dimers or triesters. It may be present in the composition as an insoluble precipitated salt of the reamer. Such salt is anio After deriving a CD with a reactive substituent, the derivatized CD is complexed with an appropriate polyvalent cation. formed by a method comprising combining or associating to form an insoluble derivatized CD salt. can do. In another or preferred embodiment, the basic elements specified above The mer structure includes novel insoluble polymeric cyclo- Contains repeating units of dextrin.

1. Cyclodextrin polymer Important?) According to a preferred embodiment, the composition comprises a derivatized cyclodextrin polymer. including. This poly "7-" is the type of inductive encyclodextrin shown in -A:/ It has a structure corresponding to a polymer formed from monomers. In light of the contents of this disclosure , it is understood that polymeric materials having such structures can be produced by a variety of methods. It will be possible. For example, derivatized cyclodextrin polymers may be The derivatized cyclodextrin monomers, dimers, trimers, etc., are combined with polymerizing agents, e.g. , epichlorohydrin, diisocyanates, diebogysides and silanes, Polymerized and/or cross-linked using procedures known in the art to form cyclodext It can be manufactured by forming a phosphorus polymer. (Insoluble cyclode Kistrin polymer beads (Insoluble Cyclodextrin Polymer Beads).

Chem, Abstr, No, 222444+r+, 102:94; Zsadon and Fenyvesi, 1st! nt. Symp, on Cyclodextrins, J.

5zejtli, ed, D, Re1del Publishing C o, Boslon, pp. 327-336, Fenibesi et al., 1979. Ann, Univ, Budapest, 5ection Chim.

15:1322. and Wiedenhof et al., 196 9. Die 5tarke21:119-123). These polymerization agents are 6.2 And the reaction can be carried out at the primary and secondary hydroxyl groups on the 3-position carbon. alternatively and preferably, first, ■ or more underivatized cyclodextrin monomers, Dimers, trimers, etc. (e.g. cyclodextrin having the structure shown in Figure 1) are combination and/or crosslinking, and then introducing anionic substituents into the resulting polymer. The derivatized cyclodextrin polymer may be prepared by: unguided cyclo Dextrin polymers consist of the epichlorohydrin linkage point of β-cyclodextrin. American Mize Products Co., Hammond, Indiana, in the form of Rimmer. It can be obtained from merican Maize Products Co. can. Commercially available underivatized polymers can be derivatized to form derivatized cyclodextrin polymers of the desired type. A remer may also be produced. The derivatized cyclodextrin polymer comprises derivatized monomers. or by reacting a mixture of underivatized monomers and derivatized cyclodext Copolymerization and/or crosslinking of phosphorus polymer and unguided cyclodextrin polymer It can also be formed by For all manufacturing operations, the polymerization method used This allows molecules to diffuse between the external solvent and a significant portion of the internal anionic monomer sites. Preferably, a solid polymer product of sufficient porosity is provided to permit permeation. .

The solubility of the CD polymer depends, among other things, on the molecular weight and size of the polymer. Dew. The present derivatized CD polymers have a large molecular weight to maintain a substantially solid state. It is a polymer of They are generally solid particles with a size of about 1 to 300 microns. It is.

The derivatized cyclodextrin polymers of the present invention are available in a variety of physical forms. and all such forms are within the scope of the invention. Suitable forms include beads, Includes fibers, resins or films. Many such polymers have the ability to swell in water. have power. The nature of the polymer product, i.e. chemical composition, degree of swelling and particle size. The distribution of Ru.

The cyclodextrin polymer derivative is preferably α, β or γ-cyclodextrin. Contains polyanionic derivatives of string polymers. Preferably (in a preferred embodiment, Anionic substituents include sulfuric acid, sulfonic acid, phosphoric acid groups, and two or more of these groups. selected from a combination of Other anionic groups such as nitrate groups have some therapeutic potential. sulfuric acid, sulfonic acid and phosphoric acid derivatives have the best therapeutic potential. It is considered to have the potential. In a preferred embodiment, fewer anionic substituents are used. At least about 10 mole percent, and more preferably at least about 50 mole percent, are sulfate groups.

Highly preferred are about 10 to 16 sulfate groups per cyclodextrin monomer. α-1β- and γ-cyclodextrin polymers containing is a tetradeca sulfate β-cyclodextrin polymer.

B, insoluble salt precipitate The composition contains induced insoluble saccharide salt precipitates, preferably induced insoluble oligosaccharides. It may also contain callide salt precipitates. As used herein, the term “salt precipitate” can be associated or complexed with suitable non-toxic physiologically acceptable cations to maintain body temperature. refers to polyanionic saccharide derivatives which form substantially insoluble salts when Ru. Suitable polyvalent molecules that can be used to produce the insoluble salt precipitate of the present invention On includes Mg, AI, Ca, La, Ce, Pb and Ba. listed here cations are generally presented in order of decreasing solubility, but different types and for saccharides with different degrees of anionic substituents this order is different. It's okay. Such induced insoluble saccharide salt precipitates are treated within the scope of the present invention. However, derivatized oligosaccharides are preferred. oligo Saccharides typically have an unsubstituted molecular weight of about 650 to about 1300.

Oligosaccharides usually have oligosaccharide fragments of a wide range of sizes. Obtained by decomposing brown rice starch or cellulose. Cyclodextrin is Generally, starch in the presence of specific enzymes favoring the formation of cyclic saccharide structures. obtained from. According to certain embodiments, the desired cyclodextrin monomer or A number of cyclodextrin monomers are added to produce the desired anionic substitution product. by reaction with a waxy substance followed by a synthetic method for the desired polyvalent type of cation. can be obtained by exchanging the cations introduced by

This latter step will precipitate an insoluble saccharide salt precipitate.

A1, Ca and Ba salts of α-1β- and γ-CD sulfates are present in the compositions of the present invention. Preferred for use, and in certain embodiments A2B-CD sulfate salts are preferred. . As is generally the case with saccharide derivatives, various Sulfation can be carried out in varying degrees. However, the derivatized cyclodextrin It is generally preferred that the salt has an average value of at least about 1.3 sulfate groups per sugar unit. more preferably at least about 2 sulfate groups per sugar unit. Particularly preferred is the group β-CD-TDS with an average value of about 2 sulfate groups per lucose unit.

C, polyanionic disaccharide derivative sucralfate (Sucratf ate) (Carafatee).

Marian Mertll Dow, Kansas C1ty, MO) is a complex salt of sucrose sulfate and aluminum hydroxide. Its structure is shown in Figure 2. . Sucralfate is α-d-glucopyranoside, β-d-fructofuranoside. roux is an octakis (hydrogen sulfate) aluminum complex.

Sucralfate is used to treat ulcers and binds to pepsin but is anti-ulcer Developed during research into weak sulfated polysaccharides. Sulfation and sucrose Its complexation with basic aluminum salts makes it a pepsin inhibitor suitable for the treatment of ulcers. Harmful molecules were introduced. Dennis M, McCa rthy), 5ucralfate, 325:14 New Eng, J , Med, 1017-1025 (1991).

Applicants have discovered that sucralfate and other polyionic derivatives of sucrose are It has some properties similar to the derivatized cyclodextrins of the present invention, with similar solubility and It has been found that it can provide affinity for growth factors. Mg, AI, Ca, La Sulfonate of sucrose in combination with polyvalent cations such as , Ce, Pb or Ba phosphoric acid or phosphoric acid derivatives bind to growth factors and direct these growth factors to the wound site. It is believed that low solubility compositions can be provided that can facilitate the therapeutic delivery of It will be done. Orally administered sucralfate has therapeutic efficacy in the treatment of gastric ulcers. Then it is written. According to the invention, sucralfate and octasulfate sulphate Other salts of loin can be pre-complexed with growth factors and physically transport the complex to the repair site. By delivering growth factor proteins to tissues or bones that need repair, It can be used for delivery.

Frequent use and/or high doses of aluminum salts pose certain health risks. It is well known that this is involved. Aluminum uptake is associated with several diseases is known or can be estimated. For example, ALLIMINUM AND H EAI, TH, A CRrTICAL REVIEW (Hillel (formerly 11el) And Gitel? :/ (edited by G i te Iman) 1 Publisher Mark Detsu Mark Decker, 1989 and A1, IJMINLIM IN 　RRNAL　FAILIIRf! .

Mark B, de Broi (Mark E, de Broi) and Jack W, Co. Jack W, Coburn, Publisher Klewer, See extensive discussion in 1990.

Aluminum is associated with osteodystrophy, osteomalacia, and mineralization disorders. It is known to cause abnormalities in bone metabolism such as m1neralization). ing. Incorporation of aluminum into the bloodstream, as can occur during dialysis, is particularly can be harmful. The following are just a few examples of the harmful effects of aluminum: This is a report. Pierides (AlM, Pierides) et al., Kidney Int, Vol, 18. 115-124.1984; Ellis (H, A , Ellis) et al., J, Cl1n, Path, 32. 832. 1979. In addition to the toxic effects of aluminum when taken into the bloodstream, aluminum salts Oral administration of also results in various adverse effects including osteomalacia and osteitis. For example, Andredi (S, P, Andredi), Bergnouutaine (J, L) Ber gstein) et al., N. Engl. J. Med, Vol. 310° 1079, 1984, Carmichael (K, A, Carmichael), F. Fallon et al., Am, J, of Med, Vol. 76 , 1137.1.984.

One of the most notable toxic effects of aluminum is that the mechanism is still unknown. Neurological abnormalities in which aluminum is speculated to play an important role, although this has not been seen before. , especially Alzheimer's disease. For example, Clapper-McLaughlan (D, R , Crapper McLachlan), Farne (B, J, Farne ll), Aluminum in Neuronal Degenerat ion, in Metal Ions in Neurology and P psychiatry.

pp, 69-87.1985. Alan R, Li5s Inc,;Bar Le (D, P, Pert), Good (P, F, Good), Uptake o f Aluminum 1nto Central Nervous System m Along Na5al-01efactory Pathways, T he Lancet, May 2゜P, 1028.1987. virtual( , 1, D, Birchall), Chappell (J, S, Chappell), Aluminum, Chemical Physiology, and Alzheimer'5 Disease, The Lancet, 0cto ber, P, 1008. See 1988.

Aluminum can be toxic, making it unsuitable for some and perhaps all therapeutic applications. In applications, non-aluminum salt forms of highly sulfated polysaccharides are Preferred over the aluminum salt form. In particular, polymers that do not require salt precipitate formation This embodiment is particularly preferred.

Some embodiments of the compositions of the invention are particularly suitable for oral administration in the treatment of gastric ulcers. Useful. In particular, non-aluminum salt-containing forms of sucrose octasulfate and most The polymeric solid form of the preferably highly sulfated cyclodextrin is It is particularly advantageous because it is free of luminium and its side effects.

(Margins below this page) D. Form of composition In view of the disclosure contained herein, those skilled in the art will appreciate that the present wound healing compositions can be used in a variety of applications. It will be appreciated that this can have beneficial effects. Therefore, for each application It is understood that, depending on the particular circumstances, the compositions of the present invention may take many and varied forms. It is illustrated. For example, the derivatized saccharide may be made into a solid gunpowder, or a dispersion or may be in the form of a suspension. Therefore, in general, compositions of the invention preferably is a derivatized saccharide and a non-toxic and physiologically acceptable compound suitable for the saccharide. Including carrier. As used herein, the term carrier broadly refers to A substance that facilitates the administration or use of this composition. Forming these compositions A variety of non-toxic and physiologically acceptable carriers can be used in It is generally preferred that these compositions are at physiological salinity.

Physically applicable for several applications encompassing wound healing in the broadest sense in a predetermined solid form that is or is implantable, the therapeutic agent of the present invention It is desirable to have a solid form of the material containing the active substance. Therefore, the original It is contemplated that the composition of the invention may be in solid form, such as a rod, needle or sheet. ing. Thus, they can be placed at or near the site of tissue damage or implantation. It may also be used externally as a wound dressing or the like. In such an embodiment, the book Compositions and compounds of the invention are preferably carried with solid carriers that are themselves biocompatible. or else the composition is suitably formed of the present saccharide derivatives. polymers or copolymers. The compositions of the invention have many applications. prepared in the form of an aqueous dispersion, suspension or paste that can be applied directly to the wound site. is preferable. To prepare these compositions, polyanionic cyclodextrin Polyanionic saccharide derivatives synthesized in solid form such as polymers is suitably purified, diluted and optionally provided with a fluid carrier such as saline. It can be used after adding other ingredients. This is a precipitate in the form of particles, The product, i.e., saccharide salt, saccharide, so that a dispersed or suspended material can be produced. This may be the case when a saccharide polymer or a saccharide copolymer is synthesized. synthesis The solid derivative is then dried, ground, or otherwise converted to the desired particle size or solid form. You can also Particle size can be optimized for the intended therapeutic use of the composition. Wear. In some preferred embodiments, the solid particles have a size of about 1 to about 600 mm. microns, more preferably about 200-600 microns. Approximately 1 to approximately 30 microns particles provide the best dispersion and rapid reactivity of growth factors. biological environment For a given weight of particles delivered to , the smaller the particle size, the larger the Ensures exposure of particle surface area so that no protein is transferred to or from the administered solid. allows for greater diffusion of active ingredients. Particles from about 30 to about ioo microns This allows for sufficient dispersion of growth factors, moderate reactivity and longer delivery of growth factors. can be obtained. Particles with a size greater than 100 microns have low reactivity, but It will provide the longest period of growth factor delivery. In certain preferred embodiments, These large particles (>100 microns) do more than deliver growth factors. It may be used in vivo to absorb blood.

In a preferred embodiment, the carrier is an aqueous medium and the composition is a solid particulate carrier. It is prepared in the form of an aqueous suspension of the tucharide derivative. The amount of induced saccharide is Preferably from about 1 to 30% by weight of the composition, more preferably from about 5 to about 15% by weight of the composition. %.

E, biologically active protein In certain embodiments, the compositions and compounds include biologically active proteins. and/or mixed with biologically active proteins. Depends on preferred embodiment For example, the biologically active protein exhibits a specific affinity for heparin, and Specifically, heparin-binding growth factors, many of which are mitogenic for endothelial cells. It is a type of growth factor that is progressive. Examples of such growth factors include basic fibroblast growth. It is a long factor. Generally, it is the satsukali of the present invention, commonly referred to as HBGF. Heparin-binding growth factor that can be mixed with (comf1ne) derivatives Probably protein. Some of these are listed in Table I.

To determine whether a protein is suitable for therapeutic compositions of the invention, has a specific affinity for heparin. H.B. The GF protein has a salt concentration that is substantially greater than the approximately 0.6 molar strength of NaCl. even in the presence of an aqueous medium with (e.g. a derivatized column) protein that remains substantially bound to heparin (using a column) It is.

Generally, the term substantially bound refers to at least approximately 80% remains bound under such conditions.

Table I IL-1 (Inter Henderson & Pettipher, 198 8゜Leukin-1) Biochem, Pharmacol, 37:417 1; Endo et al., 1988. B[1RC136:1007. Hopkin s et al., 1988〜Cl1n, Rxp, Immunol, 72:422I L-2 (InterWeiIHiIman et al., 1988.J.

Leukin-2) Biol, ResponseMed, 7:424; Gem 1o et al. + 1988. Cancer Res.

48:5864 IFNα (Inter Pitha et al., 1988. J, Immunol.

Feron α) 141:3611; Mangini et al., 1988゜Blo od　72:1553 IFN7 (Inter Blanchard & Djeu, 1988.J .

Feron 7) Immunol, 141:4067; C1eveland et al. .

1988, J. Immunol, 141:3823 TNFα (tumor necrosis Factor (Z) Plate et al., 1988. Ann, NY Acad.

Sci, 532:149; Hopkins & Meager, 1988. Cl1n, Exp.

Immunol, 73:88; Granger et al.

1988, J. Biol, Re5ponse Med.

7:48B EGF (Epidermal Growth Factor) Carpenter & Cohen, 1979 ．． A.

Rev, Biochem, 48:193-216FGF (fibroblast growth Folkman & Klagsbrun, 1987. Factors, acidity and 5 science 235:442-447 basic) IGF-1 (Insulin-like Blundell & Humbel, 198 0゜growth factor-1) Nature 287:781-787; 5choen le et al.

1982, Nature 296:252-255IGF-2 (insulin Blundell & Humbel growth factor-2) PDGF (platelet induction Ross et al., 1986. Ce1l 46:155 -169; growth factor) RiChardSOn et al., 1988. Ce1l 5 3: TGF-a (Transformation Deryck, 1988. Ce1l 54: 593-595 growth factor-α) TGF-β (transformation Cheif et al., 1987. Ce1 l 48: Growth factor-β) 409-416 The ability of heparin to complex toward growth factor proteins is Its conjugation ability and compatibility with certain cationic dye structures such as dyes and other dyes It is known to respond. Other glycosaminoglycan saccharides are similar. is known to not work. Thus, such dyes can be It has been used for many years in histology as a colorant specific for the presence of saccharides, and metachromaticity, i.e., the spectral shading that results from heparin binding on the dye. to identify active heparin-like compounds with the ability to moderate angiogenesis. has been used.

Such dye conjugation of the active protein is also carried out in the same way as the conjugation to heparin. It is highly resistant.

In relation to the present invention, it is important to note that Dye conjugation can effectively serve as an indicator of the desired activity of the compositions of the invention. It was discovered that Thus, dye conjugation analysis can be used to analyze the compositions of the present invention. Determining the ability of a polymer or copolymer to complex protein growth factors Can be done.

Heparin binding separation or chromatography process for separation of protein factors In the medium, desorption of complexed growth factors requires an additional step and very strong salt It is customary and accepted that contact with a solution is necessary.

The present invention shows that the protein conjugated to the saccharide identified here can be released at a high concentration. The important discovery and finding is that no additional step of contacting the electrolyte is necessary. This is what was used. Such operations can be performed directly as may be desired for separation techniques. As would be required for large-scale desorption methods, complex specific phases on solids and physiological surrounding fluids Low biologically required solute phase in the body relatively very A low external concentration of desorbed factors is maintained. This is a delivery agent for biomedical purposes. Applicants' compositions can be used as delivery agents. These are fundamental discoveries and findings.

Generally, to prepare a growth factor-containing composition, a growth factor or combination of growth factors may be used. A derivatized saccharide is contacted with a solution containing the combination. Then the cyclode When the cyclodextrin derivative is separated from the contact liquid, the formation on the cyclodextrin derivative is Growth factors are enriched and the fluid is correspondingly stripped of growth factors. The contact solution is , pre-isolated and pre-concentrated growth factors purified from tissues or body fluids or recombinant It may also contain one growth factor obtained by the DNA method. Moreover, the contact solution is , viable tissue or organ raw materials (hereinafter referred to as biological sources) containing various growth factors. ) may also be included. When mixed with tissue or organ materials containing growth factors , the saccharide derivatives of the present invention act as extractants for these growth factors. Can be done. If a biological source is used as the source of growth factors, the contact solution The biological source used is the body of the tissue to be treated with the mixed derivative and growth factor. has a volume greater than about to-ioo times the volume of the product.

After contact, the partially or totally complexed saccharide derivative, i.e. the solid phase , which can be easily separated from the liquid phase that is the source of the protein to be conjugated. can. The source of the growth factor is the absence of a solid other than the saccharide to be complexed. Preferably, it contains protein as a dissolved component. However, Certain solids in the long factor source solution may pose an undesirable or detrimental contaminant. It is not necessarily a dyeing substance. Is the saccharide solid, such as tissue or organ fragments? Their separation may be carried out by sedimentation, appropriate filtration, centrifugation or other mechanical or other methods. Therefore, it can be done.

■7 Methods for therapeutic regulation of wound healing One aspect of the invention is a method of therapeutic modulation of wound healing, preferably in vivo. Regarding methods of regulation, particularly regarding in vivo regulation of concentration and diffusion of protein factors. do. Such methods generally involve therapeutic in-vivo delivery of the present compositions and compounds to the wound site. Including. The low solubility of this substance, i.e. its solid immobilized state, makes it difficult to create such compositions and compounds. Allows for direct administration to the wound site and allows the active ingredient to remain at the application site for a long period of time make it possible.

Vascular cell proliferation and abnormal accumulation of extracellular matrix in the vascular wall are associated with arteriosclerosis, It is a common pathological feature found in hypertension and diabetes. Such conditions are associated with angiogenesis It is also observed after vascular injury such as surgery. Intimal hyperplasia is caused by platelet-derived growth factor ( It is thought to be partially mediated by various growth factors such as PDGF). this Growth factors act through receptors to promote vascular smooth muscle cell proliferation and from the mediators. Stimulates migration to the intima of blood vessels. Thus, Applicants have demonstrated that smooth muscle cell migration and Regulates proliferation and thereby influences the degree of intimal thickening seen after vascular injury I found a way to give it. Applicants have proposed a bovine fetal bovine protein containing strong growth factor activity. When stimulated with serum, tetradeca sulfate beta-enclodextrin stimulates human blood vessel normalization. It has been found that smooth muscle cell proliferation and migration can be inhibited in vitro.

Therefore, the presence or absence of growth factors at or near the wound site influences the healing process. It can be seen that it affects Applicants have used the present compositions and compounds to Advantageously regulates and controls biologically active proteins such as growth factors at the wound site. I found out what I can do. For example, before in vivo delivery as described here When the present compounds and compositions are mixed with a growth factor, the compositions and compounds slowly releases the offspring into the immediate vicinity of the wound, thereby accelerating the wound healing process do. All growth factors known to accelerate or promote wound healing are included in this composition. We believe that it can be used in products and methods. This formulation is suitable for accelerating wound healing. Long-term factors include those listed in Table ■, as well as brain endothelial cell growth factor and retinal-derived growth factor. Includes children. As mentioned above, heparin-binding growth factors affect both soft and hard tissues. It can be used to bring about the repair of. interferon, interloiki The potential uses of cancer and tissue growth factors are well known in the art.

The present invention provides therapeutic treatment for polyanionic saccharide derivatives or their complexes with growth factors. A method of therapeutically administering a saccharide derivative, wherein the saccharide derivative is a biocompatible porous solid. It also relates to a method of combining or including a portion of. “Bio-appropriate” used here The phrase "porous solid" refers to a substance that induces a substantial inflammatory response or other substantial side effects. means a solid that can be applied or administered to a mammal without emitting any emissions. mosquito Such biocompatible porous solids include collagen-based polymer membranes, amniotic membranes, and membranes such as the retina (details in Cobb, 1988. Eur. J, Cl1n, lnvestig, 18:321-326).

The polyanionic saccharide derivative binds the derivatized saccharide to the electrostatic bond on the membrane. immobilized on such a membrane in a preferred embodiment by contacting the sexual partner. be able to. The biocompatible porous solid is a polymer of ethylene vinyl acetate, methane Cellulose, silicone rubber, polyurethane rubber, polyvinyl chloride, polyacrylic Methyl oxalate, hydroxyethyl polyacrylate, polyethylene terephthalate, Polypropylene, polytetrafluoroethylene, polyethylene, polyfluoroethylene Also tyrene, propylene, cellulose acetate, cellulose and polyvinyl alcohol (Including details: Hoffman, 5ynt, hetic Poly meric Biomaterials in PolymericMa,te reals and Artificial Organs, AC3Symp osium 5eries #256゜(G, Gebelein ), 1988), in a preferred embodiment, the cyclodextrin starting material is copolymerized with monomers of the biocompatible polymeric material of the final product composition to create a porous Make a copolymer. This copolymer is then chemically reacted to form a compound according to the invention. Carry out the saccharide moiety with the required anionic substituents. biocompatibility Reactions such as amine, amide, carboxylate end groups, etc. contained in polymers Coupling of functional groups with cyclodextrins followed by derivatization with ionic substituents. Ru. More preferably, a polysaccharide such as a cyclodextrin is added to a solid polysaccharide. Introduced as a co-reagent in the monomer formulation to be polymerized into a polymer or copolymer, followed by This product is then contacted with a suitable reagent to induce the saccharide component. Anionic substituents are added to the extent taught by the inventor. Such methods and Particularly advantageous are the products manufactured by 3M Company for use in biocompatible patch or wound preparations. is a flat polymer product of polyamide polymer used as a dressing. A method of producing example polymers or copolymers.

This biocompatible patch or dressing provides physical protection to the wound from pathogen invasion. It is made to have enough pores to allow moisture, air, etc. to pass through.

Applicant's invention describes the use of activated polyanionic polysaccharides in combination with such polymers. or the active anionic saccharide. and the protein factor together with the polymeric carrier. Intended. Such combinations may be used to intentionally promote or inhibit cell growth processes. clearly intended for use. HBGF is incorporated into or already present in biological membranes. A component whose basic structure is an immobilized induced saccharide, which is either Join to child. Biological membranes such as the retina or amniotic membrane are used in the art as wound dressings. It is well known. Synthetic biofilms whose main component is collagen are used to treat burns. Ru. The presence of the derivatized saccharides of the present invention in neutral membranes such as amniotic membranes and synthetic membranes. The ability of these derivatives to bind to collagen used as a support is an advantage of the present invention. Such biomembranes can be used as novel delivery vehicles for HBGF when combined with compositions. He will make it possible for you to stay.

A. Restenosis Arteriosclerosis is a disorder that involves thickening and hardening of the walls of large blood vessels in mammals. It is a major cause of arterial disease, aortic aneurysm, and lower extremity artery disease. Arteriosclerosis is caused by cerebral It also plays an important role in vascular diseases.

Until now, angioplasty has been a widely used method to treat arteriosclerosis. Ru. For example, percutaneous transluminal coronary angioplasty (hereinafter referred to as "PTCA") In 1988 alone, it was held over 20,000 times in the United States. The PTCA method is Inject the deflated balloon catheter through the skin into the narrowed blood vessel or artery. Including inserting into. The catheter is then inserted into the blood vessel until the narrowed area is reached. Opens into the cavity. This area of stenosis consists of fatty streaks, fibrous tissue that narrows the blood vessel and restricts blood flow. It is characterized by the accumulation of sexual plaques and associated lesions on the vessel walls. Arteriosclerosis To overcome this harmful artery narrowing caused by the condition, a balloon is inflated. to collapse the plaque against the artery wall, and then use other methods to expand the artery lumen. It is.

Although PTCA provides excellent results and low complication rates, the use of this method requires It is difficult. In particular, the enlarged artery wall will cause the balloon to inflate against the artery wall. Frequently suffers damage and injury during

Although this injury itself does not appear to be particularly harmful to the patient's health or life, The healing response caused by the injury can relapse the atherosclerotic condition. In particular, arteries In response to direct or inflammatory injury to the artery, smooth muscle cells associated with the narrowed area of the artery It is recognized that it initiates a division. Smooth muscle cells proliferate and grow inside arteries As they migrate into the layers, they cause thickening of the arterial wall. Initially, this thickening increases This is due to smooth muscle cells. However, after that, further thickening of the arterial wall and and narrowing of the lumen due to increased smooth muscle cell volume and extracellular matrix and connective tissue. This is due to the accumulation of Thickening of this cell wall and narrowing of the lumen after atherosclerosis treatment is referred to here as restenosis.

Applicants are not bound by any theory or theories regarding the basic principles of restenosis. Although unwanted, restenosis is the overgrowth of smooth muscle cells exposed to endothelial injury. partially due to the presence of growth factors produced by the injured endothelium that activate It is thought that

Accordingly, Applicants have proposed that the present saccharide derivatives are suitable for use with growth factors prior to in vivo delivery. Substantially free of vascular angioplasty may prevent or reduce intimal thickening after balloon angioplasty. It has been found that it is extremely effective in reducing, at least substantially, to growth factors By virtue of their affinity for such growth factors, such compositions may vo absorption or local concentration reduction and/or diffusion. That is, These wound site growth factors are dependent on whether they are produced by cells at the wound site. whatever is present in the bloodstream can be removed by the present saccharide derivatives, thereby This reduces the restenosis effect of such materials on the injured tissue.

According to this method, mammals, including humans, having arterial regions suffering from arteriosclerosis can The polyanionic saccharide derivative of the present invention is administered to mammals to induce arterial smooth muscle cell proliferation. treated by administering an amount effective to inhibit. The degree of restenosis inhibition , depending on factors such as the patient to be treated and the extent of arterial injury during the angioplasty procedure. It is contemplated that variations may be made within the scope of this specification. However, the saccharide One condition is that the derivative is administered in an amount effective to result in a substantial reduction in restenosis. Generally preferred. As used herein, the term substantial reduction in restenosis refers to This means that the restenosis value is not greater than about 50%. According to a preferred embodiment, the treatment Post-treatment restenosis values are not greater than about 25%. When used here, the post-treatment restenosis value and The term refers to restenosis values measured approximately one month after angioplasty. Restenosis value The term ``increase'' means greater than 50% of the initial increase in minimum lumen diameter obtained by angioplasty. The restenosis rate is calculated as the threshold or equivalent loss.

Thus, the present invention provides a method of inhibiting restenosis in a patient, the method comprising: saccharide in an amount effective to inhibit the formation of restenosis in patients who have undergone A method is contemplated comprising administering to the patient a derivative thereof. The satsuka ride The derivative may be administered before, during and/or after angioplasty treatment of narrowed arteries. It is considered that it may be administered to administering the compound locally to the wound site. The method of administration is generally preferred. In a preferred embodiment, topical administration comprises involves injecting the id derivative directly into the injured tissue. In case of restenosis, The step preferably involves injecting the compound directly into the artery wall at the angioplasty site. including doing.

Applicants have surprisingly found that particularly beneficial anti-restenotic results are due to the saccharide-induced the step of administering to the body comprises expanding the lumen of the blood vessel to perform angioplasty; For example, Applicants have found that a favorable (7) The step is to apply an aqueous suspension or dispersion of Saracara 1 derivative to the balloon angioplasty site. 1, which preferably involves injection into the arterial wall. is a partially modified infusion catheter with multiple holes in the wall of the balloon portion of the catheter. It is performed using a balloon catheter.

These holes allow inflation solution to seep out from the walls of the balloon when the balloon is inflated. It is arranged and made to such a size.

According to a preferred embodiment, the balloon is inflated at a relatively low pressure, such as 2 to 3 atmospheres. Masu. Porous balloon cap that can be used to apply the composition of the invention Examples of teters are u, s, c, +, -]<- Bard and Shu. This is done by Sehneider. This type of balloon , Wolinsky/<Runo or "sweating balloon" be exposed. For application of the composition of the present invention, various injectable angioplasty balloons may be used. A catheter can be used, and one skilled in the art will know which type of catheter with an infusion balloon. It can be seen that it is easy to determine whether the tell is appropriate. Saccharide induction of the present invention Another method involving local administration of the body uses bioabsorbable intravascular stents. It is something. Saccharides of the invention, especially cyclodextrin polymers/derivatives into a bioabsorbable stent and place the stent at or near the site of tissue damage. It may be located nearby.

The individual characteristics and properties of the suspension containing said saccharide derivatives do not necessarily depend on the invention. It is understood by those skilled in the art that the It will be understood by the person. However, the administration step preferably An aqueous suspension or dispersion of ionic saccharide derivative particles, preferably about 1 to 6 A suspension of 00 micron sized sulfated β-cyclodextrin polymer particles was , involves injection directly into the balloon angioplasty site within the artery. Applicants: Such particles instilled into the arterial wall are applied in sufficient quantities to cause inhibition of restenosis. It remains in the area for several days (I think it will continue for 7 days).

The aqueous suspension is composed of an aqueous carrier of physiological salinity. including. The active saccharide derivative preferably comprises about 1 to about 30 parts of the composition. % by weight, more preferably from about 5 to about 15% by weight. Preferred embodiment odor The derivatized tucaride, preferably siflovkis 1-phosphorus sulfate polymer particles. is generally applied during angioplasty.

In some instances, it is desirable to prevent restenosis but allow angioplasty. It can be said. To meet these requirements, polyanionic saccharide-derived A1 or Ba salt of body, more preferably AI of polysulfated β-cyclodextrin Alternatively, it is preferable to use a dispersion of Ba salt. Inhibiting restenosis and vascular injury at the same time If it is desired that normal angiogenesis proceed at the site, Sucralf From Marian Merrill Dow Co., Kansas City, Missouri. Preferably, the aluminum salt of sucrose octasulfate, which is readily available, is used.

B. Inhibition of intimal thickening of vein grafts. Venous segments are frequently harvested during surgery and are used to treat vaso-occlusive disorders. Used as an Ipus graft. Specifically, they include, for example, coronary, renal, large The femoral and axillary arteries are used in the circulatory system. One of the main uses of this form of treatment is The limitation is that intimal thickening occurs, which weakens the luminal cross section and reduces flow. be. This occurs frequently, but not exclusively, with anastomoses. Applicant et al. introduced β-cyclodextrin tetradeca sulfate into the perivascular space during surgery. The placement of polymer particles substantially limits smooth muscle cell ingrowth into the vascular intima. We propose that this would improve the long-term success rate of these grafts.

C1 angiogenesis Angiogenesis is the formation of new blood vessels. Angiogenic stimulation induces endothelial cell elongation and proliferation. and the generation of new blood vessels. Some HBGFs can promote angiogenesis Are known. New blood vessels generated by angiogenesis lead to tissue vascularization .

There are various diseases that involve insufficient blood supply to tissues and organs. This is known as ischemia. Deficiencies in species may be due to functional constriction of blood vessels or actual obstructions. These diseases Diseases are grouped into cardiac, cerebral and peripheral ischemic diseases. Cardiac ischemia is chronic It can result in angina or acute myocardial infarction. Cerebral ischemia can lead to stroke .

Peripheral ischemia can result in a number of diseases, including arterial embolism and frostbite. Peripheral In severe cases of ischemia, occluded blood vessels cause tissue necrosis and require amputation. imaginary To overcome blood pressure, another blood supply to the injured tissue must be established. do not have.

According to a preferred embodiment, angiogenesis is carried out by first forming a su-tucharide derivative of the invention. the composition is then placed in the ischemic tissue, e.g. by subcutaneous injection. Promotes angiogenesis and collateral blood vessel formation by local administration to . As used herein, the term collateral vessels does not exist under normal physiological conditions. (＼ refers to blood vessels that appear in response to appropriate stimuli such as the presence of HBGF.) Administration of a composition comprising a tucharide derivative and a growth factor can reduce collateral vessel formation and ischemia. It is thought to bring about revascularization of sexual tissue.

In a preferred embodiment, angiogenesis is induced by a highly anionic cyclodextrin derivatives or salts thereof, more preferred (Ba is a cyclodextrin derivative or a salt thereof) Facilitated by methods involving polysulfated polymers or copolymers. That shiku The lodextrin derivative and the basic fibroblast growth factor were mixed at a ratio of about 10:l-100: Mix the weight ratio of cyclodextrin:basic fibroblast growth factor to 1. is preferred.

D0 tissue and organ transplantation As mentioned above, HBGF is known to stimulate angiogenesis and endothelial cell growth. There is. In transplantation, the success of the transplant procedure depends on whether the transplant procedure is re-wounded or not. , is critically dependent on the speed of establishment of an adequate blood supply to the transplanted tissue. mosquito Thus, Applicants apply the composition of the invention mixed with growth factors to the transplant site. The aim is to promote the establishment of an adequate blood supply to the transplanted tissue. growth factor The child-containing composition may be coated onto the surfaces to be joined or sprayed onto the surfaces. or in the form of an aqueous suspension with or without thickening agents such as glycerol. May be applied. In addition, the organ or tissue to be transplanted may be treated with a composition containing the composition of the present invention. It may be pre-soaked before implantation into a treatment solution. The composition of the present invention is bonded to Both implant sites or surfaces may be injected.

Preferred methods for preparing compositions used to treat transplanted tissues and organs In the method, the saccharide composition of the present invention is combined with a growth factor-containing biological source (e.g. For example, tissue or organ debris, ground matter, or liquid extracts. The growth factors present within these sources are extracted. emergency In a preferred method, the biological source used for contacting is treated with the composition. It is about 10 to about 100 times larger in volume than the implant. more direct and more economical A common method is to produce the saccharide derivatives of the invention by recombinant biochemical and biological methods. may involve contacting with a growth factor substance produced by otechnological manipulation. . In this way, specific growth factor proteins for the intended therapeutic application can be Quality is easier to choose.

E. Bone grafting Bone responses to insults such as fractures, infections, and interruptions in blood supply are relatively limited. Ru. In order for damaged bone tissue to heal, dead bone must be broken down and resorbed, and new bone must be formed. must be formed, and a process induces the growth of new blood vessels within the area involved. The accompanying process must be carried out. HBGF promotes angiogenesis and osteogenic cells. Can induce proliferation. Therefore, fracture healing and the connection between the grafted bone and the host bone are and to promote bone mineralization (where such is intended). It is intended that the compounds be used in combination with growth factors.

In a preferred embodiment, the present saccharide derivative is used as a growth factor and powdered bone material. and/or finely dispersed demineralized bone to form a paste. mosquito A suitable method for preparing such a paste is Repair of Major. Cranio-Orbital Defects with anElasto mer Coated Mesh and Autogenous Bone Pa5te, Mutatsu B.

Habal (Mutaz B, Habal) et al. 61:3. Plastic and ReCOReC0n5trLICtiVeSur, 394. ．． 39 6 (1978). The bone tissue used to make the paste is from the intestines. It can be obtained from the bony crest or calvaria.

It is preferable to use autologous bone for transplantation purposes, as it is preferable to use bone powder that has been completely demineralized. Preferably, partially demineralized bone is used.

To prepare the bone meal, demineralized bone meal obtained from allogeneic and xenogeneic sources is used. May be used. To make a soft paste absorbent cellulose, use cotton or similar material. May be used. Applicants do not wish to be bound by any theory or theories. Although undesirable, the bone paste produced by these methods has a network of blood vessels. It is thought that it functions as a guiding matrix from which new bone is generated after formation. It will be done. The paste may be applied to the surface of the bone to be joined in a grafting procedure or is used to fill the contour bone of the fracture to be repaired.

F. Skin ulcer treatment Millions including but not limited to aging, paraplegics, trauma victims and diabetics One debilitating disorder affecting people is non-healing skin ulcers or pressure sores. Ru. In many cases, insufficient blood supply to injured tissue results in adequate nutrients for healing. is preventing the delivery of Compounds such as epidermal growth factor and basic fibroblast growth factor Polymer beads of nclodextrin derivatives are preabsorbed by mixing with ulcers. By applying directly to the skin, angiogenesis is increased, blood supply is improved, and keratinosis It is believed that site ingrowth will increase and ulcer closure and healing will be faster.

G. Dermatological applications Control of blood vessel growth is an important aspect of normal and pathological conditions encountered in dermatology. be. In particular, abnormal growth of cellular material and blood vessels is associated with several pathological conditions. Drsoriasis is 1 Here are two major examples. Often involves an excess of growth-stimulating protein factors . This type of abnormality is often accompanied by an imbalance of protein growth factors.

For example, in patients with cutaneous mastocytosis, extracts from the affected skin may be The source may be an unrelated skin or control sample from a patient without such deficiency. 15 times higher level of chymotrybutin activity (Chymotryp tic activity) (Human 5kin Chymot ryptic Protease, Schefter (N, M.

5chechter), Fraki (J, E, Fraki), Giesin (J, C, Geesin).

Lazarus (G, S, Lazarus), 1.8io1. Chem... 258.2973-2978°1983). The related chymase is Heparin-binding factors (S, Sayama), ions (R, V, Iozzo), Lazarus (G, S, Lazarus), Schefter (N, M , 5chechter), l(t+man SkinChymotryps in-1ike Proteinase Chymase, J, [1io1 ．． Chem, 262゜6808-6815. (see 1987).

Cymoto-lipsin-like proteases degrade epidermal junctions and cause epidermal-skin separation. (See Sayama et al. above).

Another example of a growth-promoting factor associated with skin abnormalities is epidermal plasminogen activity. It is an embodiment. This increases in various skin abnormalities (Epidermal Plasminogen Activator is Abnormal 1n Cutaneous Legions’, Jensen (P, J, Jensen ) et al., J., Invest.

Dermat, 90-777-782.1988).

Certain embodiments of the invention, i.e., highly sulfated solid dispersions or highly sulfated other physical variants of polyzaracarides, preferably cyclodextrin structures Particularly applicable to skin treatments when the body contains overgrowth of cellular components. Ru. In such cases, the substance of the present invention is introduced into the relevant tissue or its vicinity. .

This makes it suitable for dermal or subcutaneous injection or effective contact of finely divided dispersions of the substance. can be carried out by implantation of a solid polymer molding shaped to In preparations such as batch preparations or other external preparations in a suitable form containing the substance of the present invention, May include substances.

Depending on the pathological and disease state, pre-contacting the substances of the invention with protein growth factors It will be understood that it is intended to be applied without. This is a In situations such as those exemplified above, where one seeks to reduce any growth-promoting factor or factors, Probably.

In other cases of skin damage or skin diseases, and in certain stages of treatment, proteins It may be desirable to use a mixture of factors. This is called angiogenesis, or new There may be cases in which the establishment of a blood supply and additional blood supply is desired in conjunction with the healing process.

(Leaving space below) Example The following examples are presented to illustrate the invention. However, those should not necessarily be construed as limiting the scope of the invention; The scope of is determined by the appended claims. All amounts and percentages shown are Weight basis unless otherwise specified.

Example 1 Preparation of sulfated β-cyclodextrin polymers of 20-60 mesh particle size β-cyclodextrin polymer beads (American Mize Products) derivatized to form a novel immobilized sulfate CI) polymer derivative according to the present invention. Applicable The composition approaches the degree of sulfation of the CD polymer in glucose or almost di-sulfuric acid.

About 0.4 g of carefully dried polymer is mixed with about 1.7 g of 6-trimethylammonium. um sulfur trioxide complex (Aldrich) and approximately 100 ml of dry dimethylformamide. The reaction was carried out for 3 to 4 days at approximately 62 to 72°C with gentle stirring in DMF (DMF). . The solid was washed in DMF and reacted with 30% aqueous sodium acetate for 24 hours. , washed and stored in distilled water. The sulfur content of the product is approximately 14.7% by weight. It was. If the polymer mass contains 100% Te1-radeca sulfate β- It is composed of cyclodextrin and all glucose hydroxyl units are erected. If it is physically available (which is not expected for the polymer), this is a problem. It easily compares to the value of 1.7.5%.

Example 2 (A) β-CD-TDS (Na): β-cyclodextrin (99% purity dihydrate) from Chemalog (Chemalog, South Plainfield, New Jersey). malog) (a branch of General Dynamics).

Approximately 5.0 g of β-cyclodextrin (approximately 4.4 mmol, or approximately 92 mmol) -0H) was dissolved in about 250 ml of dimethylformamide (DMF). child Add about 15 g of (CH3)3N-3o3 (about 108 mmol) at once to the solution of The reaction mixture was heated to about 70°C. After 2 hours at about 70℃, the rubber A substance started to precipitate. The reaction mixture was maintained at 70°C with vigorous stirring and then and cooled to room temperature. The DMF layer was then decanted and discarded, leaving a solid residue. was dissolved in about 250 ml of water and then about 75 ml of 30% sodium acetate was added. Ta. The mixture was stirred vigorously for 4 hours and then poured into approximately 4000 ml of ethanol. I got into it.

After one night of incubation, the mixture was filtered to collect the crystallized solid.

The filter paper cake was washed with ethanol (anhydrous) followed by diethyl ether. . The product was then dried under vacuum over P2O.

Approximately 10.3 g of white powder was collected. The product was hygroscopic.

The products were analyzed under conditions that minimized water sorption. The following results were obtained from elemental analysis. was: C=18.84, H=2.65, S=17.33 (CgHeOzS Theoretical value for zNa2; C=19.67, H-2,19,5=17.49) . [α: 1D22=75° (c=2.63.0° in 5M NaCl). The analysis value is 2 hydroxyl groups for each glucobylanose unit, i.e. 1 per CD molecule. This corresponds to the estimated average degree of substitution of 4 hydroxyl groups. Such β-CD- The theoretical yield of TDS salt is 96 g of lO, about 6% higher than the measured value of 10.3 g.

(B) a- and 7-CD-3 (Na salt): approximately 86 mmol of (CH3 )3N-3o, and the above procedure was used, except that about 117 mmol of γ-CD was used. was used for these preparations.

Sulfated α-CD salt was analyzed as C = 18.76; H = 2.60.3 = 16.22 It was done. This translates to an average of approximately 11.7 hydroxyl units of substitution per α-CD molecule. Corresponds to degree.

The sulfated γ-CD salt has C=18.92, H=2.69, and 3=14.84. was analyzed. This is an average degree of substitution of about 14 hydroxyl units per γ-CD molecule. corresponds to

(C) β-CD-30, (Na salt) (7.1% by weight S): Approximately 1.0 g of β-CD-30, (Na salt) (7.1% by weight S) The lodextrin was dissolved in approximately 50 ml of DMF. Approximately 883 mg of CH,N 803 (7.2 eq.) was added. The solution was heated to about 75°C for about 12 hours. No precipitate formed during this time. Cool the reaction mixture to room temperature did. Approximately 200 ml of ethanol was added to the solution. Next 0, the generated colloid The solution was poured into about 600 ml of diethyl ether. White solid for 2 hours It was generated in the middle of the day. The solid was collected by filtration and then dissolved in approximately 30 ml H2O. It was dissolved in This solution was stirred for 2 hours. After stirring, the solution was diluted with about 900 ml of 2: Poured into IEtOH-EtzO solution. Crystals were formed over 8 hours. the crystal were collected and washed with EtzO. The product was dried in a vacuum oven with P2O5. Approximately 1.1 8 g of powder was collected (72.4% yield).

Elemental analysis of the product showed C=32.49.8=4.99, and S-7,06. Indicated. This corresponds to an average degree of substitution of approximately 3.5 hydroxyl groups per β-CD molecule. handle.

(D) β-CD-propoxylate-x4sO+β-CD-(hydroxy-n- propyl ether) from American Mize Products (Hammond, sulfate, i.e., β-CD-(~4 propylene 14S04) was prepared.

Example 3 Preparation of growth factors Received human recombinant basic fibroblast growth factor (bFGF) from Takeda Pharmaceutical Co., Ltd. Ta. It was purified from E. coli as previously described (Kurokawa okawa) et al., 1987. FBBS, Letters213:189- 194 and Iwane et al., 1987. Biochem, mouth 1 op hys, Res, Commun, 146:470-477).

As previously described (Shing et al., 1984°5ci ence 223:1296-1298), rat chondrosarcoma-derived growth factor (C hDGF) was isolated from transplantable tumors. prepared by collagenase digestion of the tumor. Approximately 100 ml of the prepared crude extract was diluted with approximately 0.6M NaC in approximately 10mM Tris, pH 7. Dilute with 1. :1), heparin-Sepharose pre-equilibrated with the same buffer Packed onto a column (1,5×9 cm). About 0 in about 10mMh of pH7 ．． The column was washed with approximately 100 ml of 6M NaCl. Next, about 10 m of pH 7 ChDGF was eluted with approximately 18 ml of approximately 2M NaCl in M Tris.

Example 4 β-cyclodextrin affinity chromatography insoluble sulfation of FGF - Measure the cyclodextrin polymer (approximately 0.5 ml volume) and unit of human recombinant bFGF (about 10 mM) at pH about 7) in squirrel ．． Ink with about 0.5 ml of 1M NaCl while mixing at about 4℃ for about 1 hour. It was incubated. Subsequently, about 0.1, 0.6, and The polymer was washed stepwise with approximately 2 ml each of 2M Nacl and 2M Nacl. from the polymer All fractions were analyzed for growth factor activity.

Example 5 growth factor analysis Quiescent state of BAL B/c mouse 3T3 cells in 96-well Brake 1 Measuring the amount of thymidine incorporated into the DNA of the dense layer of Growth factor activity was evaluated by: ■The activity of the unit is within 3T3 cells (approximately i 50% maximum DNA synthesis in o, ooo cells (10.25 ml growth medium/well) It was defined as the amount of growth factor required to stimulate growth. To measure specific activity For this purpose, we analyzed the crude extract and the active fraction eluted from the heparin-Sepharose column. The protein concentration was determined by the method of Lowry et al. (1952, J.

Biol, Chem, 193:265-275). Pure Protein concentrations of growth factors were determined by silver-stained polypeptides on 5DS-polyacrylamide gels. The intensity of the peptide band was evaluated by comparing it with that of the molecular weight marker.

Example 6 Fibroblast growth factor on tetradeca sulfate β-cyclodextrin polymer Affinity Human recombinant bFGF (approximately 1000 units) was added to β-cyclodextrin sulfate. Incubated with polymer. Subsequently, the polymer was diluted with about 0.1M, 0. It was eluted stepwise with 6M and 2M NaCl. The results are shown in Figure 3.

Most of the growth factor actives remained bound to the polymer at about 0.6M NaCl. However, when eluted with about 2M NaC], about 230 units of active substance were recovered. child These results indicate that basic fibroblast growth factor is FGF has a very strong affinity for heparin, and has a very strong affinity for heparin. It shows that they are comparable. The activity peak is 5DS-polyacrylamide gel. The samples were analyzed by silver staining after electrophoresis. Lane 2 in Figure 4 shows salt The basic fibroblast growth factor polypeptide band is shown.

Heparin and tetradeca sulfate β-cyclodext against chondrosarcoma-derived growth factor Phosphorus affinity was also tested. Cartilage meat containing about 500 units of active growth factors The tumor extract was transferred to heparin-Sepharose and β-cyclodextrin tetradeca sulfate. were incubated separately with the polymer. Subsequently, the beads were heated to about 0.1. Stepwise elution was performed with 0.6 M, 0.6 M, and approximately 2 M NaCl. The results are shown in Figure 5.

Approximately 32% and 68% total activity was observed for heparin-cephalin at 2M NaCl, respectively. 1 and tetradeca sulfate β-cyclodextrin polymer.

匡 molecular weight 30.000 1. ,l□ /4,300■ 1 noon 123 FIG.4

Claims (46)

[Claims] 1. Polyanionic with a body temperature solubility of less than about 15 g per 100 ml of distilled water Saccharide derivatives and physiologically acceptable carriers for the saccharide derivatives A composition for influencing the growth of living tissue in a mammal, comprising: 2. 2. The composition of claim 1, further comprising a heparin-binding growth factor. 3. The saccharide derivative contains an average of at least about 1.4 anilines per sugar unit. 2. The composition of claim 1, having an onic substituent. 4. The saccharide derivative has an average of about 1.4 to about 4 anions per sugar unit. 4. The composition of claim 3, having a sexual substituent. 5. The saccharide derivative is a compound consisting of n sugar units and an R anionic substituent. It is a compound, When n = 2 to 3; average anionicity R ≧ 3.5 per n unit, when n = 4 to 5 If: average anionicity per n unit R≧2.0, if n≧6; average anionicity per n unit 4. The composition of claim 3, wherein the average anionicity R≧1.4. 6. The saccharide derivative contains a polyanionic cyclodextrin derivative. The composition of claim 1. 7. 7. The composition of claim 6, wherein the cyclodextrin derivative comprises a compound of the formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ (wherein at least two of the R groups per monomer unit are sulfuric acid, phosphoric acid, sulfuric acid, selected from the group consisting of phonic acid and nitric acid, and the remaining R groups, if present, are This is H, alkyl, aryl, ester, ether, thioester, thioether is a non-anionic group selected from the group consisting of L and -C00H, where n is about 6 to about It is an integer of 12. ) 8. The cyclodextrin derivative has an average of about 10 anions per monomer. composed of one or more cyclodextrin monomers having a sexual substituent, The composition of claim 6. 9. The monomers contain an average of about 10 to about 24 anionic substituents per monomer. 9. The composition of claim 8, comprising: 10. The cyclodextrin derivative comprises a cyclodextrin polymer. The composition of claim 7. 11. the polymer is solid particles dispersed or suspended in the carrier; The composition of claim 10. 12. The cyclodextrin derivative may be a polyanionic α-, β- or γ-cyclodextrin derivative. 7. The composition of claim 6, comprising a salt of clodextrin. 13. The cation components of the salt include Mg, Al, Ca, La, Ce, Pb, Ba and and a combination of two or more of these. 12 compositions. 14. 2. The composition of claim 1, wherein said derivative is substantially insoluble in water at body temperature. 15. at least a portion of said saccharide derivative is dispersed in said carrier or suspended solid particles. 16. A method for suppressing abnormal growth of smooth muscle cells in a mammalian tissue, the method comprising: A polyanionic saccharide derivative is added to the tissue to inhibit the abnormal growth. topical administration in an effective amount, such that said derivative is administered topically in an effective amount of about 1 ml per 100 ml of water at body temperature. A method having a solubility of less than 5g. 17. 17. The method of claim 16, wherein said derivative is a cyclodextrin derivative. 18. The anionic substituent of the cyclodextrin derivative is a sulfate group, a sulfone group, Essentially consisting of acid groups, phosphate groups, nitrate groups, and combinations of two or more of these 18. The method of claim 17, wherein the method is selected from the group consisting of: 19. the cyclodextrin derivative is the polyanionic cyclodextrin derivative; 20. The method of claim 18, further comprising a salt of . 20. Mixing said derivative with a non-toxic, pharmaceutically acceptable carrier of physiological salinity. 20. The method of claim 19. 21. Claim wherein the derivative comprises a sulfuric acid derivative of β-cyclodextrin polymer. The method of Section 16. 22. A solid in which said derivative is dispersed or suspended in a physiologically acceptable carrier. particles, the suspension or dispersion comprising from about 1 to about 30% by weight of the derivative; 17. The method of claim 16. 23. 12. The suspension or dispersion comprises from about 5 to about 15% by weight of the derivative. 22 methods. 24. 17. The method of claim 16, wherein said derivative is mixed with a growth factor. 25. The local administration comprises injecting the saccharide derivative directly into the tissue. , the method of claim 16. 26. Said topical administration comprises administering an aqueous suspension or dispersion of said saccharide derivative to said saccharide derivative. Using a catheter with an injection balloon that has multiple holes in the wall of the balloon part, 17. The method of claim 16, comprising injecting directly into the tissue. 27. Polyanionic saccharide inducer of cyclodextrin mixed with growth factors Promoting angiogenesis in a mammal comprising locally administering a conductor to the tissue to be treated 2. A method, wherein the derivative is present at body temperature in an amount of less than about 15 g per 100 ml of distilled water. A method characterized by its solubility. 28. 28. The method of claim 27, wherein said derivative is a cyclodextrin derivative. 29. The anionic substituent of the cyclodextrin derivative is a sulfate group, a sulfone group, Essentially consisting of acid groups, phosphate groups, nitrate groups, and combinations of two or more of these 29. The method of claim 28, wherein the method is selected from the group consisting of: 30. the cyclodextrin derivative is the polyanionic cyclodextrin derivative; 30. The method of claim 29, comprising a salt of 31. Mixing said derivative with a non-toxic, pharmaceutically acceptable carrier of physiological salinity. 31. The method of claim 30. 32. Claim wherein the derivative comprises a sulfuric acid derivative of β-cyclodextrin polymer. The method of Section 27. 33. A solid in which said derivative is dispersed or suspended in a physiologically acceptable carrier. particles, the suspension or dispersion comprising from about 1 to about 30% by weight of the derivative; 28. The method of claim 27. 34. 12. The suspension or dispersion comprises from about 5 to about 15% by weight of the derivative. 33 methods. 35. The local administration comprises injecting the saccharide derivative directly into the tissue. , the method of claim 29. 36. Said topical administration comprises an aqueous suspension or dispersion of said saccharide derivative. Assemble using a catheter with an injection balloon that has multiple holes in the wall of the balloon part. 28. The method of claim 27, comprising injecting directly into the tissue. 37. A composition comprising a polyanionic derivative of cyclodextrin and a growth factor. Transplant tissue in mammals, including contacting the organ or tissue to be transplanted or a method of treating a transplanted organ, the composition comprising: Administered in an amount effective to promote acceptance, the derivative is administered per 100 ml of water at body temperature. The method has a solubility of less than about 15 g. 38. The contacting step comprises locally administering the derivative to the tissue or organ. , the method of claim 37. 39. A method of treating injured or transplanted bone, the method comprising: mixing bone tissue with growth factors; Compositions containing polyanionic derivatives of cyclodextrin that have been contacting the derivative in an amount effective to promote bone healing in the body, A method having a solubility of less than about 15 g per 100 ml of water at room temperature. 40. 40. The method of claim 39, wherein the composition further comprises bone tissue. 41. 41. The method of claim 40, wherein the bone tissue comprises pulverized bone. 42. 41. The method of claim 40, wherein the bone tissue comprises finely dispersed demineralized bone. 43. 41. The method of claim 40, wherein the composition is a paste. 44. 41. The method of claim 40, wherein the bone tissue comprises autologous bone. 45. 41. The method of claim 40, wherein the bone tissue comprises xenogeneic bone. 46. Method of producing compounds suitable for influencing the growth of living tissue in mammals A cyclodextrin polymer is prepared, the cyclodextrin polymer is mer with an anionic derivatizing agent in the presence of dimethylformamide at about 62 to about 72°C. for about 3 to about 4 days at a temperature of less than about 15 g per 100 ml of water. The polyanion of the cyclodextrin polymer characterized by the solubility of A method comprising obtaining a chromogenic derivative.