MXPA97007634A - Cyclodextrine compounds and methods of manufacturing and using the mis - Google Patents

Abstract

The present invention relates to: polyanionic, substituted cyclodextrins having cell growth modulation activity. The invention also provides cyclodextrins having anionic groups on one side of the cyclodextrin molecule. Therapeutic methods of use, as well as methods for making the cyclodextrin compounds of the invention are also described herein.

Description

CYCLODEXTRINE COMPOUNDS AND METHODS OF MANUFACTURE AND USE OF THEM 1. FIELD OF THE INVENTION This invention relates to highly anionic cyclodextrin compounds having additional substituents with hydrophobic properties attached. More particularly, it is also directed to physiologically acceptable compositions comprising these compounds, and to methods of use for modulating cell growth activity. 2. BACKGROUND OF THE INVENTION 2. 1 HEPARINE AND ITS CELLULAR MODULATION PROPERTIES It has become increasingly evident that there is a class of biological activities of heparin, which is distinct from those of the classical anticoagulant properties that have long been associated with heparin. This involves different types of interaction with protein factors, and the modulation of proliferative behavior and other behaviors of cells. Heparin interacts chemically by forming complexes with growth factor proteins (heparin binding growth factors, HBGF's), such as FGF (see, for example, Y. Shing, J. Folkman, R. Sullivan, C. Butterfield, J. Murray, and M. Klagsbrun, Science 223, 1296-1299, 1984, M. Klagsbrun and Y. Shing, Proc. Nati, Acad. Sci. USA 82, 805-809, 1985). It modulates the metabolism of cells in many ways (see, for example, A. Yayon, M. Klagsbrun, J.D. Esko, P. Leder, and D.M. Ornitz, Cell 64, 841-848, 1991). It protects growth factors such as FGF against proteolytic degradation (see, for example, D. Gospodarowicz and J. Chen, J. Cell Physiol., 128, 475-484, 1986); O. Saksela, D. Moscatelli, A. Sommer, D.B. Rifkin, i. Cell Biol. 107, 743-751, 1988); it promotes endothelial cell proliferation and angiogenesis (see, for example, J. Folkman, R. Langer, R. Linhardt, C. Haudenschild and S. Taylor, Science 221, 719, 1983; LB Castellot et al., J. Cell Physiology 127, 323, 1986; T. Barzu et al., J. Cellular Physiol., 140, 538, 1989; SC Thornton, SN Mueller, EM Levine, Science 222, 623-625, 1983; SN Mueller, KA Thomas , J. DiSalvo, and EM Levine, J. Cell Physiol., 149, 439-448, 1989, DB Volkin, PK Tsai, JM Debora, J.0.Gress, CJ Burke, RJ Linhardt and CR Middaugh, Arch. Biochem. and Biophys., 300, 30, 1993; S. Taza, Y. Hayakawa, T. Ishikawa, K. Niiya, N. Sakuragawa, Thrombosis Research 72, 4, '31 -439, 1993). Heparin also inhibits the proliferation of soft muscle cell (A. Clowes and M. Karnovsky, Nature 265, 1977, J. Guyton, R. Rosenberg, A.

Clowes and M. Karnovsky, Circ. Res. 46, 625, 1980; J. Castellot Jr., i. i. Choay, J.C. Lormeau, M. Petitou, E. Sache and M.J. Karnovsky, i. Cell Biol. 102, 10979, 1986)). In conjunction with steroidal agents and other agents, heparin inhibits angiogenesis (J. Folkman, R. Langer, R. Linhardt, C. Haudenschild and S. Taylor, Science 221, 719, 1983, b) R. Crum, S. Szabo and J. Folkman, Science 230, 1375, 1985; J.K. Lee, B. Choi, R.A. Sobel, E.A. Chiocca, R.L. Martuza, J. Neurosurg. 73, 429, 1990). It also provides other functions of altering the cellular behavior, such as inhibiting the infection of the HIV virus (M. Ito, M. Baña, A. Sato, R. Pauwels, E. DeClercq, S. Shigeta, Antiviral J? S. , 361-367, 1987), and others. Unfortunately, one is limited with respect to the use of heparin in a variety of therapeutic applications of the types indicated above due to mainly two reasons: one is the fact that there is no strict uniformity in the chemical composition between different heparin preparations. Thus, it was shown that the efficacy of heparins of different origins, when used to inhibit angiogenesis, varies from good to very poor (see J. Folkman, R. Langer, R. Linhardt, C. Haudenschild and S. Taylor, Science 221, 719, 1983). On the other hand, one is limited in the use of heparin by the fact that heparin inevitably introduces the anticoagulant property, which at the doses indicated for many of the potential applications would lead to complications such as bleeding, stroke, et cetera. It seems that the compositional and structural characteristics of heparin, required for the cell modulation activities described above, are different from those required for their anticoagulant activities. We have sought to reduce the complexity of the structure of heparin to the simplest chemical composition required for the class of cell modulation properties, and which, at the same time, would be expected to be more compatible with the living tissue environment. We have succeeded in obtaining a comparable or greater modulation behavior after the total abandonment of the flexible, long and complex chain structure of heparin glycosaminoglycan, in favor of a simple and rigid toroid of only six to eight glucose units, namely cyclodextrin (CD), if we provide a high molecular density of anionic substituents such as a minimum number of sulfate groups, as described in U.S. Patent Number 5,019,562 of MI Folkman and P.B. Weisz This type of composition, such as a cyclodextrin polysulfate (CDS), is a "heparin mimic" that was found to control angiogenesis (J. Folkman, PB Weisz, MM Joullie, WW Li and WR Ewing, Science, 243 , 1490, 1989), to inhibit soft muscle cell proliferation, promote the production of endothelial cells in culture, and inhibit the invasion of HIV virus from human cells (PB Weisz, HC Herrmann, MM Joullie, K. Kumor, MS Levine, EJ Macarak, D. Weiner, Angiogenesis and GAG-Mimics, in Angiogenesis - Key Principies - Science-Technology-Medicine, R. Steiner, PB Weisz, R. Langer, Eds, Birkhaeuser-Springer, Basel and New York, 1992; D.B. Weiner, W.V. Williams, P.B. Weisz, M.I. Greene, Pathobiology 60, 206, 1992). Precisely as it is well known in the science and study of heparin, sulphated cyclodextrins interact with cells through adsorption on the membranes of the cell, as is evident by their protective cells from virus invasion (see Weiner et al. , above) and for its protection of erythrocytes against destruction by hemolytic agents (PB Weisz et al., Biochem. Pharm. 45, 1011-1016, 1993). 2. 2 CICLODEXTRINS AND THEIR USE IN PHARMACOLOGY Cyclodextrins (referred to hereinafter, for convenience, as CDs or CDs for the singular and the plural, respectively) are cyclic oligosaccharides consisting of at least six units of glucopyranose. Although CDs with up to twelve glucopyranose units are known, only the first three homologs have been extensively studied. These compounds have the simple, well-defined chemical structure shown in Figure 1 (A). The common designations of the lowest molecular weight OI-, ß- and? -CDs are used throughout this specification, and will refer to the chemical structure shown in Figure 1 (A), where n = 6, 7 or 8 glucopyranose units, respectively. The initial discovery of CDs with starch degradation products was made by the turn of the century, and Schardinger showed that these compounds can be prepared by the action of Bacillus macerans amylase on starch. In older literature, compounds are often referred to as Schardinger dextrins. Sometimes they can also be called cyanoamylose. Topographically, the CDs can be represented as a torus, as shown in Figure 1 (B), whose upper edge is delineated with primary -CH20H groups, and the lower edge with secondary hydroxyl groups. Aligned coaxially with the torus is a channel-like cavity with a diameter of approximately 5, 6 or 7.5 A.U. for a-, jS- and? -CDs, respectively. These cavities make cyclodextrins capable of forming inclusion compounds with hydrophobic host molecules of suitable diameters. A reasonably large number of cyclodextrin derivatives have been prepared and described in the literature. In general, these chemically modified cyclodextrins are formed by the reaction of the primary or secondary hydroxyl groups attached to carbons 2, 3 or 6 [Figure 1 (A)], without disturbing the bonds or; (l--> 4) hemiacetal. In "Tetrahedron Report Number 141, Synthesis of Chemically Modified cyclodextrins", A.P. Croft and R.A. Bartsch, Tetrahedron 39 (9): 1417-1474 (1983), incorporated herein by reference for background (hereinafter referred to as "Tetrahedron Report No. 147"), a review of these preparations is given . In particular, the sulfates of o; -, β- and y-cyclodextrin (Na salt) are shown as Compounds Nos. 207, 208, and 209 in Tetrahedron Report No. 147, (supra) Table 26, page 1456. The U.S. Patent No. 2,923,704 to Berger describes the preparation of cycloamylose sulfates. U.S. Patent No. 4,020,160 to Berstein et al., And U.S. Patent Nos. 4,247,535 and 4,258,180 to Lewis et al. Describe the use of modified cyclodextrin sulfates as complementary inhibitors. U.S. Patent No. 4,383,992 to Lipari discloses the preparation of a water-soluble inclusion compound of a steroid and an unmodified 0-cyclodextrin. U.S. Patent No. 4,596,795 to Pitha discloses the administration (via the sublingual or buccal route) of sex hormones, particularly testosterone, progesterone and estradiol, in the form of their inclusion compounds with hydroxypropyl-β-cyclodextrin or poly - | S-cyclodextrin. With respect to the applications of cyclodextrins to the field of pharmacology, there is a growing number of applications concerning the solubilization of different pharmaceutical compounds. This is based on the ability of the toroidal structure of the cyclodextrins to internally accommodate a large variety of molecules through the internal formation of complexes, with the proviso that (1) the host molecule or a significant portion of it can be physically molded and pass through the toroidal opening, and (2) is sufficiently hydrophobic or lipophilic to be maintained there by the non-covalent interaction with the equally hydrophobic structure of the internal atomic sugar skeleton of the cyclodextrin. This complexation has been used to internalize a wide variety of pharmaceutical agents and, in this way, to apply them within an aqueous solution when these can not be dispersed in another way within the solution, due to their low solubility. In addition, this internal complexation has been used for agents that have haemolytic activity. The examples of the prior art are the inclusion of cyclodextrin of phenothiazine neuroleptics (see Otagiri et al., Proc. Of the First Int. Symp. On Cyclodextrins, pages 3893981 1982; Uekama et al., J. Pharmacodyn. 4,142-144, 1981), steroid hormones (see Ukama et al, Int.J. Pharm.10, 1-15, 1982), antiinflammatory and analgesic agents (see Lister et al., Eur. J. of Rheu. And In. fl airan., 12, 6-11, 1993, antihypoxia drugs (see Wallerstein and Cserhati, J. Biochem and Biophys. Methods, 29, 49-60, 1994), opioids such as morphine, fentanyl and others (see Jang et al. , J. Pharm. And Exper. Therap 261, 592-600, 1992) and many more, although different cyclodextrins and cyclodextrin derivatives have been described and described, for a growing number of these applications, it should be noted that the technique it does not make use of highly ionic cyclodextrins It is believed that none of the above references show or make obvious the invention of the applicant, as described and further claimed later. 2. 3 SIGNIFICANCE AND APPLICATIONS OF THE METHODS FOR MODULAR CELL PROLIFERATION A large number of biological processes are caused by and directly related to events that involve or lead to a lack of, or overexpression of, cell proliferation. Obviously, in the embryonic development, cell division and proliferation to a controlled degree is essential. Similarly, any reconstructive or healing processes, after the destruction of tissue or organs, either by injury or due to pathological causes, demands the recreation of cells and cellular materials. In this way, the induction or improvement of cell proliferation is desired, in relation to the wound healing processes. Similarly, in ischemia, due to the loss of active blood vessels or capillary cells for the growth and formation of endothelial and new cells and the endothelium. Cellular growth is also sought in the practice of surgery, such as in the practice of implantation, where the growth of cell junctions is important, and the accommodation and growth of new blood vessels and capillaries for acceptance, and where it is It is probable that the accelerated accommodation decreases the immune rejection. It has been shown that the foreign introduction of the growth factor protein is useful (see Roberts and McGeachie, J. Anat., April 169, 197-207, 1990, Stagner and Samols, EXS 61, 381-5, 1992) and when it can be practiced in a controlled manner. It has become clear that growth factor proteins play an important, if not decisive, role in all these constructive (ie, embryonic) and reconstructive processes. In contrast to the previous cases, where the induction of cell proliferation is important, natural and excessive proliferative processes rarely occur in the adult body, except as a result of injury or disease. Particularly prominent is the phenomenon of neovascularization, or the induction of neovascularization, known as angiogenesis. This involves the proliferation of endothelial cells that subsequently form new capillaries and blood vessels. Many diseases are either caused or accompanied by angiogenesis. Tumor growth requires constant elongation of the nutrient, i.e., the blood supply system. Active growth, therefore, requires angiogenesis. The inhibition of angiogenesis can, therefore, inhibit or terminate tumor growth. In certain diseases of the eye, uncontrolled growth of blood vessels through the cornea can occur; or this one can destroy the retina. Both conditions can lead to blindness, if angiogenesis is not controlled. Similarly, angiogenesis accompanies rheumatoid arthritis in joints, in psoriasis, and other dermatological conditions and pathologies. Therefore, there is a need for means and methods to control angiogenesis, that is, to control endothelial cell proliferation. Another example of cell proliferation is that which follows the almost inevitable damage of the superficial layer of the blood vessels, which occurs with angioplasty, or which accompanies the accumulation of plaque in cardiovascular disease. These conditions lead to the proliferation of soft muscle cells, which can lead to restenosis after the angioplastic intervention, or to a more rapid increase in the size of the material, leading to more rapid obstruction of the flow. Again, the need to inhibit cell growth under these conditions is relevant and urgent. 2. 4 THE NATURE OF POLYANIONIC CYCLODEXTRIES (HIGHLY SULFATED) PREVIOUSLY DISCLOSED The usual sulfation methods (for example, the use of chlorosulfonic acid, trimethylamine / sulfur trioxide complex, or sulfur trioxide / pyridine complex) that were practiced in the prior art, by others (for example, see United States Patent Number 12,923,704 of L. Berger; Patent of the United States of America Number 4,258,180 of A.J. Lewis) or by us (United States Patent No. 5,019,562 to Folkman and Weisz, and the numerous references to the authorship of Weisz with Folkman, Joullie, Weiner, Barnathan, Macarak, and others, in Section 2.1 above) to the substitution of sulphate on both sides of the cyclodextrin toroid. The sulfate groups are positioned in any of the positions (hydroxyl) designated by 2, 3, or 6 of the glucose rings of the cyclodextrin structure. Figure 2 illustrates this. All publications referenced above are incorporated herein by reference. 3. COMPENDIUM OF THE INVENTION This invention is directed to substituted polyanionic cyclodextrins (CDs) and their uses. Cyclodextrins having both multiple and anionic polar substituents, as well as multiple non-polar substituents of substantial hydrophobic or lipophilic character, are provided for optimal use in the treatment of various pathologies. Preferred cyclodextrins are referred to as "one-sided" anionic forms of the highly anionic substituted cyclodextrin, wherein most of the anionic substituents are located on one side of the cyclodextrin toroid structure. The water solubility of the compounds of the present invention can be conveniently changed from insoluble to highly soluble by varying the carbon chain length of the non-polar substituents. Other aspects of the present invention are compositions containing the aforementioned compounds, for use in many therapeutic methods, methods having to do with the modulation of cell growth activity, as discussed below. The invention also provides compositions for inhibiting cell proliferation of soft muscle cells, comprising (1) highly anionic substituted cyclodextrins having a sufficient number of hydrophobic substituents and (2) a physiologically acceptable application medium, wherein the degree of the hydrophobic character of the substituents provides improved absorption within the blood plasma, depending on the mode of application to mammals, including humans. These modes of application include intravenous or oral application, injection into body fluids, tissue or organs, or topical application. The invention also provides compositions for promoting endothelial cell growth, comprising (1) highly anionic substituted cyclodextrins bearing hydrophobic substituents and (2) growth factor protein, wherein the degree of hydrophobicity of the substituents is likewise adjustable. The invention also provides compositions for the inhibition of neovascularization and angiogenesis, comprising (1) highly anionic substituted cyclodextrins having hydrophobic substituents and (2) a steroidal or non-steroidal angiostatic compound, wherein the degree of the hydrophobic nature of the substituents it is provided to achieve improved absorption within the blood, as described above. The invention further provides angiogenesis inhibiting compounds which are highly anionic substituted cyclodextrins having hydrophobic substituents, wherein the angiostatic compound is included as at least one of the hydrophobic substituents attached to the cyclodextrin or, alternatively, is part of a complex of inclusion with a "one-sided" polyanionic cyclodextrin. On the other hand, the invention conveniently provides compositions which are inclusion complexes of highly anionic substituted cyclodextrins and pharmaceutical or therapeutic agents of limited solubility in water. By the same, this invention provides methods for the solubilization of a variety of pharmaceutical agents in a manner that allows greater amounts of the agent to be applied than in the prior art. Therefore, this surprisingly provides a more effective application of the therapeutic agent included directly to the surfaces of the cell or tissue, than has been achieved so far. The invention also provides methods of application which, with respect to the number of polyanionic groups and the length of the hydrophobic substituent (s) of the compounds of the invention, optimize the efficacy and dose index requirements, for achieve the desired therapeutic goal. In addition to the solubilization and application of the pharmaceutical compounds of interest, the inclusion complexes of the present invention also allow the stabilization of these decomposition compounds. By providing the compounds of this invention with different degrees of hydrophobicity (lipophilic), this invention allows improved methods of application. For example, depending on the nature of the target tissue, more lipophilic compounds are especially suitable to provide greater affinity to fatty tissue. Also, the ability to lower the solubility in body fluids can help in the location of the compounds, that is, in management applications. On the other hand, hydrophobicity aids the penetration of cell membranes, and conveniently increases the absorption of oral ingestion within the bloodstream. The invention also conveniently and surprisingly provides, through the use of the compounds of this invention, methods of treating many pathologies, which involve abnormal cell proliferation activity, such as hypercholesterolemia, and the prevention of stenosis or restenosis in cardiovascular disease, or after angioplasty, as well as in angiogenic diseases such as corneal neovascularization, retinopathy, tumor growth, rheumatoid arthritis, psoriasis and other pathologies accompanied or caused by angiogenesis. Therapeutic methods of the invention also encompass the positively modulating cellular activity in such a way that wound healing, transplantation and ischemic tissue revascularization are improved. Another aspect of the present invention is a method for making the substituted polyanionic cyclodextrin compounds. Conveniently it has been found that the compounds possess a minimum critical number of polyanionic groups, that critical number being at least ten (10). The compounds of the invention can also be conveniently synthesized in such a manner to be of a desired water solubility, whether low, intermediate, or elevated, based on the carbon length of a hydrophobic substituent attached at the hydroxyl positions of the compound of cyclodextrin, which are free of anionic groups. More particularly, the present invention provides a method for making the hydrophobically substituted polyanionic cyclodextrin compounds, wherein the polyanionic groups are positioned substantially on one side of the cyclodextrin. By way of summary, the present invention conveniently and unexpectedly provides polyanionic, hydrophobically substituted cyclodextrins, particularly cyclodextrins which are anionically substituted on "one side" of the molecule, which is beneficial and surprisingly suitable for application in a wide variety of therapeutic methods. The pharmacological activity of the compounds of the present invention allows a modulation method of cell behavior, whose cellular modulating activity includes a) promoting endothelial cell proliferation, as occurs during the process of implant / transplant surgery, or wound healing; b) inhibit endothelial cell proliferation when in combination with an angiostatic compound such as a steroid, or a retinoid, whose angiostatic activity is adequate to inhibit tumor cell growth, or to inhibit neovascularization in different angiogenic diseases, such as retinopathy or psoriasis; c) inhibiting soft muscle cell proliferation, for example, under different cardiovascular conditions, such as inhibiting restinosis after angioplasty; and d) promote membrane stability of cells and tissues, by preventing viral infection of cells, preventing the hemolysis of erythrocytes by different hemolytic agents, and avoiding glomerular membrane leakage in nephrology and diabetes, or leakage of protein in interstitial cystitis and inflammatory bowel disease. In accordance with the foregoing, the compounds of the present invention are conveniently and unexpectedly suitable for use in a wide variety of therapeutic applications, whose use meets the long-stated needs described above. 4. BRIEF DESCRIPTION OF THE FIGURES The present invention can be understood more fully by reference to the following detailed description of the invention, the examples and specific embodiments of the invention and the figures in which: Figure 1 (A and B) shows a schematic representation of (A) the chemical structure of beta-cyclodextrin, and (B) the three-dimensional, toroidal structure of it. Figure 2 shows the toroidal representation of the anionic (sulfated) cyclodextrin, in which any of the possible substituent positions (2, 3, and 6) carries the anionic substituent (sulfate). Figure 3 shows a summary of the accumulated data concerning the critical dependence of the biological activities of cell modulation, based on the number of anionic groups (sulphate) per molecule of polyanionic cyclodextrins, for antiangiogenesis, promotion of endothelial cell growth, inhibition of soft muscle cell growth, inhibition of virus invasion in T cells, and optical metachromatic activity, which is a biochemical indicator of cell-related biological activity (see AC Grant et al., Anal. Biochem. 137, 25-32, 1984), and PB Weisz et al., In Angi ogenesis - Key Principle is - Sci enc - echnol ogy - Medi cine, Steiner Weisz, Langer, Eds, Birkhaeuser - Springer, 1992). Figure 4 (A, B, C, D) shows examples of the "one-sided" anionic cyclodextrins of this invention. Figure 5 shows the synthetic steps to obtain a cyclodextrin polysulfate with a steroid (hydrocortisone) as a lipophilic and pharmacologically active substituent of the polyanionic cyclodextrin.

. DETAILED DESCRIPTION OF THE INVENTION The present invention provides CD compounds (CD) polyanionic, substituted, having a critical minimum number of linked polyanionic groups. The invention provides for at least 10 anionic groups per CD molecule. Such critically substituted CD compounds have been determined to be suitable for use in multiple therapeutic applications, as will be explained below. The compounds of the present invention possess cell growth that modulate properties similar to that of heparin, while conveniently lacking the undesirable anticoagulant activity of heparin. With respect to the anionic groups in the CD molecule, these can be the anions of any strong acid. Non-limiting examples of these anions include sulfate, nitrate, sulfonate, and phosphate. Sulfate is preferred. Furthermore, it has been conveniently discovered that the solubility in water of these compounds can be varied from low to intermediate to highly soluble by varying the length of the carbon chain of a non-polar substituent attached to the polyanionic cyclodextrin, the substituent of which is substantially hydrophobic or lipophilic. By low solubility is meant virtually insoluble or from 0 grams / 100 milliliters to approximately 0.5 grams / 100 milliliters; by intermediate solubility is meant from about 0.5 to about 25 grams / 100 milliliters; and by highly soluble is meant a solubility of at least about 25 grams / 100 milliliters, preferably at least about 30 grams / 100 milliliters. All solubility determinations were carried out at 0 ° C. Suitable non-limiting examples of non-polar substituents, which are substantially hydrophobic or lipophobic, include an alkyl, an aryl, an ester, an ether, a thioester and a thioether. These substituents can be varied relative to the length of their carbon chain to achieve a suitable or desirable water solubility of the substituted polyanionic cyclodextrin. By varying the length of the substituent group, one can alter the solubility of the CD molecule, depending on the desired therapeutic application. In accordance with the above, a substituent of hydrophobic chain length greater than about 20 gives a polyanionic CD, substituted of low solubility; a substituent length of from about 7 to about 20, provides a cyclodextrin of intermediate solubility; and a length of less than about 7, results in a cyclodextrin, which is highly soluble in water. In any case, it is considered within the skill of the art to vary the carbon length of the substituent groups for the purpose of achieving the desired water solubility of the CD compounds of the present invention. With regard to the number of hydrophobic substituents that can be attached to the CD molecule, this number can range from about one to about 14. This provides the critical minimum number of anions of at least about 10 per CD, and regardless of whether the cyclodextrin it is alpha, beta or gamma. For example, the number of hydrophobic substituents that may be attached may range from about one to about 8 for alpha-CD; from about 1 to about 11 for the beta-CD; and from about one to about 14 for the CD-range. These cyclodextrin compounds can be substituted in any available hydroxyl groups of the CD molecule by an anion or a non-polar substituent, provided that the critical minimum number of anionic groups is given at least ten. Figure 1 describes the structure of the beta-CD (A) and a more three-dimensional view of it which shows its toroidal structure. Figure 2 illustrates a sulfated CD. Figure 3 shows that there is an acute criticality in the number of anionic (sulfate) groups required for cellular modulating activity, namely, a requirement of at least about 10 anionic groups. The high density of the anionic charges around the CD, while clearly providing a highly desirable cellular biological activity, could also generate some limitations of use. It is known that the permeability of the cell membrane and the tejidq, for example, is inhibited by the charge density and is favored by a more hydrophobic (lipophilic) character. Therefore, an entirely ionic compound limits certain aspects of transportation in the body, such as penetration into the blood plasma of oral application, penetration of the blood-brain barrier, and so on. There are also other desirable biochemical activities resulting from hydrophobic structures, such as affinity for association with and capture and transport by the same of other hydrophobes, such as cholesterol, carotenes, and others. This invention therefore provides CDs that possess the required critical anion density, while also possessing hydrophobic (lipophilic) portions to conveniently provide other desirable properties, such as increased blood absorption. Also as shown in Figure 3, the compounds of the present invention having a critical minimum number of anionic groups of at least about ten (10) exhibit significant antiangiogenic activity in combination with hydrocortisone; promotion of endothelial cell proliferation; inhibition of soft muscle cell proliferation; and inhibition of viral infection of the cells by HIV-1. It is understood that the solubilities of the compounds of the present invention will be altered if hydrophilic groups are introduced into the hydrophobic chain, as exemplified by Figure 4D, thereby increasing the length of the chain that would otherwise provide more solubility. low. . 1 CDS WITH CRITICAL ANION DENSITY AND HYDROPHOBIC SUBSTITUTES In the prior art, substituted cyclodextrins with the density of the critically elevated anionic group or the additional non-ionic substituents have not been described, but are described in our Application of the United States of America with Number No. 07 / 947,417, which is incorporated herein by reference, where non-ionic substituents including short chain alkyl substituents such as methyl, ethyl, n-propyl, and isopropyl groups were mentioned, which retained a substantially high solubility of sulfated CDs, while modifying groups such as ester, ether, thioester, thioether, and carboxylate can add hydrophilic activity. The invention provides substituted CDs with the density of the critically elevated anionic group, and additional nonionic substituents of hydrophobicity high enough to substantially reduce the water solubility of the hydrophobically substituted polyanionic cyclodextrins. Accordingly, the compounds of this invention do not carry groups of nonionic substituents containing non-polar atom chains (of carbon or sulfur atoms) of at least three atoms, or more, if additional polar modifier groups are also employed. as substituents. If the provided number of non-polar substituents is smaller than it would correspond to all positions of. hydroxyl not substituted by anions, then longer hydrophobic chains would be required. It is therefore more convenient to define the total degree of hydrophobicity provided by the resulting reduction in water solubility achieved. Therefore, it will be the objective to provide a non-polar substitution sufficient to achieve a reduction in water solubility of at least 30 percent. Those compounds of the present invention that have a low solubility in water are particularly suitable for therapeutic applications that require the localization and retention of the compound at a desired site. Such compounds provide limited dispersion of the compound by diffusion, while conveniently remaining at the desired therapeutic treatment site. By way of the present invention, one of skill in the art can decrease the solubility of the compounds by covalently linking the hydrophobic substituents to the polyanionic CD molecule. Another method for decreasing the solubility of the compounds is by the formation of much higher molecular weight entities from monomeric compounds described herein. This can be done by oligomerization or polymerization of the cyclodextrin before the synthesis of the substituted derivatives. In fact, the polymerization link can be achieved by polymer binding agents containing long chain hydrophobic groups themselves. In the United States Patent Number 5,262,404 the polymerization methods of CD monomer are mentioned and referenced, which is incorporated herein by reference. . 2 CDS WITH CRITICAL ANION DENSITY AND HYDROPHOBIC SUBSTITUTEANTS ADDING ANOTHER BIOLOGICAL FUNCTIONALITY The invention additionally provides an additional biological functionality to the cellular adhesion and cellular modulating capacity of the substituted polyanionic cyclodextrin. This is achieved by choosing at least one of the hydrophobic substituents, a compound known to have pharmacological activity. Non-limiting examples are steroids, angiostatic steroids, bactericides, or antibiotic agents, and a variety of other agents such as antioxidants, for example, beta-carotene. In this way, the addition as a substituent of a steroid such as hydrocortisone, provides a compound that can inhibit angiogenesis, the substituted compound of which possesses the heparin-like activity of the cyclodextrin polysulfate, as well as that of the angiostatic steroid hydrocortisone. It is understood that an angiostat is a latent growth inhibitor that does not possess any by itself or that has a negligible antiangiogenic activity, but requires heparin or a heparin mimic such as CD-polysulfate to achieve that activity. Other steroidal or non-steroidal structures, which are well known to one of skill in the art, can also be employed. Most preferred are those latent growth inhibitory steroids lacking glucocorticoid and mineralocorticoid activity, since such activity is an undesired effect, and limits the amount of dose or extent of use of the steroid for the purpose of the present invention. Among these most preferred steroids are 11 alpha, 17, 21-trihydroxyprg-4-en-3, 20-dione (or 11 alpha-hydrocortisone), 17 alpha, 21-dihydroxyprg-4-en-3, 20-dione ( -desoxipregna-4, 9- (11) -dien-3, 20-dione The term "cortisone" and "hydrocortisone" and 11-a isomer of hydrocortisone is intended as used in the present specification, and in the claims, includes both steroids themselves and their derivatives and structural variants thereof None of the latent growth inhibitory steroids themselves effectively inhibit angiogenesis, nor does it cause regression of tumors in the absence of a highly soluble substituted cyclodextrin sulfate in Water of the present invention, associated with a physiologically acceptable cation Further, in the methods of the present invention any non-steroidal organic compound can be used, which in combination with a substituted CD sulfate associated with a physiologically acceptable cation table, demonstrate growth inhibitory activity in any of the bioassays described below. As taught by the present invention, the growth inhibitory activity of the non-steroidal organic compounds is made potent by the combination with a water-soluble substituted CD sulfate, associated with a physiologically acceptable cation. Among the organic compounds that can be used in the present invention are the following: proline analogues such as L-2 azetidinecarboxylic, cis-hydroxyproline, and 3,4-dihydroproline and trans-retinoic acid and other retinoids. Many well-recognized bioassays have been developed to estimate the angiogenic inhibition potency if any, of a substance. The rabbit cornea is the basis of one of these methods. The cornea is avascular. A small pocket can be made in it, and a tumor implant inserted while the rabbit is anesthetized. The tumor is separated from the vascular bed of the host. New capillary blood vessels will grow in a linear fashion toward the tumor, and the rate of vessel growth can be measured. For a more detailed description of this trial, see, Gimbrone et al., J. Nat 'l Cancer Inst. 52: 413 (1973) incorporated herein by reference. A more economical bioassay makes use of the chorioallantoic membrane of the chicken embryo. For convenience, hereinafter referred to as "CAM trial". For a more detailed description of the CAM assay, see Folkman et al., Science 221: 719 (1983), incorporated herein by reference. A typical CAM assay employs 16 eggs per experiment. A disc of 2 millimeters in diameter is applied to the chorioallantoic membrane of a 6-day-old chicken embryo, grown in a Petri dish in a humidified incubator with 3 percent carbon dioxide. Two days later (8-day embryo), the membrane is examined under a stereomicroscope at six to ten times amplification. The inhibition of angiogenesis by the test substance is made evident by the development of an avascular zone around the methylcellulose disk. To an avascular area of 4 millimeters it is qualified as (++), to an avascular zone of 2 millimeters it is qualified as (+). The power of the inhibition in the zone (s) of 2 millimeters and 4 millimeters is expressed as the percentage of the total number of eggs (usually 10) in the test that were evaluated (++) or (+), is say, the percent inhibition of the test substance under the test conditions. The sustained release methylcellulose disks are prepared by dispersing the appropriate amount (s) of the test substance in a 0.45 percent aqueous solution of methylcellulose, and depositing 10 microliter aliquots of the solution resulting in a Teflon mold, followed by air drying for about one hour in a laminar flow hood. A very convenient feature of the CAM assay is the very high sensitivity of the chicken embryo to toxic substances. In addition, the lack of toxicity of a substance in a CAM assay has been correlated well with the lack of toxicity of that substance when it is administered to other animals. . 3"SINGLE SIDE" CYCLODEXTRINS WITH CRITICAL DENSITY OF ANIONS In addition, it has now been conveniently and surprisingly discovered, by means of the present invention, that the substituted polyanionic cyclodextrin compounds, described above, wherein the anionic groups are substantially linked only in positions 2- and 3- of the cyclodextrin molecule, unexpectedly possess superior biochemical and pharmacological properties. Hereinafter these are referred to as "one-sided" cyclodextrins. These "single-sided" cyclodextrins are particularly suitable for use in the different therapeutic applications described herein. Therefore, the present invention conveniently provides cyclodextrins having a critical density of anions, which possesses cellular modulating activity, wherein the anionic substituents, for example, sulfates, are located primarily on one side of the toroidal structure of the cyclodextrin, while the other side of the toroid has substituent groups large or entirely non-ionic, non-polar, and preferably hydrophobic, if any. Referring to Figure 1 and Figure 2, it should be noted that there are certain restrictions on the available number of substituents on either side of the cyclodextrin toroid: with the beta-CD, there are seven positions 6 on one side, and fourteen positions 2- and 3 in the other. It is clear, therefore, that to achieve a total of at least ten anionic substituents (eg, sulfate) in the cyclodextrin molecule, this number can not be accommodated on the side of the 6-position alone. Most or all anions should be placed, therefore, on the side of position 2- and 3-. In accordance with the foregoing, this invention, when it involves the "one-sided" polyanionic cyclodextrin compounds, provides compounds wherein most or all of the anionic substituents are placed at the 2-or 3- positions. It will be noted that the mere replacement of the 2-or 3- positions alone would yield at least seven anions, well beyond the critical requirement for cell modulation activity. The "single-sided" cyclodextrin compound of this invention also allows flexibility with respect to addition in the number and nature of substituents on the 6-side side, thereby providing the ability to alter the degree of hydrophobicity . The hydrophobicity determines the solubility in water of the molecule, the ability to interact with other hydrophobes in solution, or on the surfaces of cells or tissues, the ability to effect and modulate the penetration capabilities of biological membranes, and also the capacity to choose as aggregate substituents, compounds that possess an additional pharmacological activity, for purposes and in the same way as previously discussed. . 4 COMPLEXES OF INCLUSION WITH CHLORODEXTRINES OF CRITICAL DENSITY OF ANIONS Many years of study involving cyclodextrins lead to the hypothesis that any cyclodextrin that has the familiar hollow cavity in its toroidal structure will harbor guest molecules of adequate size. These inventors also made this hypothesis when describing in the United States Patent Number 5,019,562 (M.J. Folkman and P.B. Weisz) the polyanionic cyclodextrin, and beta-cyclodextrin tetradecasulfate, to perform the inhibition of angiogenesis. However, we subsequently discovered that it is not possible to obtain the inclusion within the highly polyanionic substituted cyclodextrin. After consideration of the dimensions and locations of the toroidal structure of the cyclodextrin, and the diameter of the anionic substituent groups, it becomes clear that the anionic groups were located outside, at the entrances of the toroidal structure. Its large number required for biological activity, together with the large diameter of each group, for example the group (-0-S03) -, causes the entrances to the cavity to be blocked due to steric hindrance. The present invention, therefore, unexpectedly provides biologically active cyclodextrins, which have the critical number of anions required, however surprisingly capable of forming inclusion complexes with molecules or portions of molecules that conform to the size of the cavity, and possess sufficient hydrophobicity to interact in an attractive manner with the hydrophobic nature of the internal cavity of the cyclodextrin. This is indeed surprisingly accomplished by the "one-sided" anionic cyclodextrins of this invention, since these compounds possess the desirable concentration of high anion density on one side of the cyclodextrin toroid, while leaving the entry on the other side No obstruction by bulky anionic groups. This conveniently provides the ability to form inclusion complexes that surround those polyanionic cyclodextrins, with an almost unlimited variety of possible host molecules having the required hydrophobicity. Suitable molecules include partially or fully hydrophobic (lipophilic) organic compounds, such as, for example, alkanes, alkenes, aromatics, fatty acids, lipids, terpenes, and biological and pharmacological agents, such as hydrophobic or partially hydrophobic steroids, vitamins, hormones. , antioxidants, such as retinoids, bactericides, antibiotics and antiviral agents, particularly anti-HIV agents, such as AZT and ddI, or other nucleoside derivatives. We have also found that although the inclusion of molecules of pharmacological interest within the polyanionic cyclodextrin of the prior art is unfeasible, an inclusion complex can be created by means of the present invention. For example, we succeeded by the same, to obtain the internal complex between the cortisone and the sulphated cyclodextrin, as shown by the spectroscopic absorption of the product. . 5 MULTIPLE BIOLOGICAL FUNCTIONS POSSESSED BY COMPOUNDS AND COMPOSITIONS The compounds and compositions of this invention conveniently possess multiple biological functions and pharmacological activities, as compared to the cyclodextrin compounds that include the sulfated compounds (polyanionic) of the prior art. In accordance with the above, the compounds and compositions of the present invention are especially suitable for use in the therapeutic methods described below in Section 6. Polyanionic cyclodextrins, such as heparins, form external electrostatic complexes between their negative anions and the multiple groups cationic (basic) proteins, such as growth factor proteins, also known as heparin binding growth factors (HBGFs). These are, as we noted, "heparin mimics". As a result, polyanionic cyclodextrins are bound, as in the case of heparins, to protein and protein surfaces, attaching themselves to the cell and tissue surfaces by virtue of their strong electrostatic forces. By the addition of substantially hydrophobic substituents, the cyclodextrins of this invention can also interact with other biologically significant lipophilic groups. By providing added hydrophobic groups that are themselves pharmacologically active, novel pharmacological properties are also provided. By providing the anionic group on only one side of the toroidal structure of cyclodextrin ("one-sided" anionic cyclodextrins), we provide, in addition to the existing electrostatic fixability due to the high density of anions, the capacity of generate inclusion complexes with other agents, agents that in this way can be solubilized, stabilized, and applied in a more efficient way than previously achieved. Although the application by use of cyclodextrin inclusion complexes per se is a widely developed technique, the compounds of this invention conveniently combine the strong adsorption properties in cells and tissues, thereby surprisingly increasing direct application capabilities more there simply depending on the desorption of the host molecule and the diffusion of the target surfaces.

By the same, many variants are enabled in the methods of application and adaptation to different tasks of therapy or biotechnology, as discussed later in the present. 6. APPLICATION AND METHODS OF USE The compounds of the present invention are especially useful in a multiplicity of pharmacological and medical applications. These constitute significant improvements in the applications described previously, such as the use of anionic cyclodextrins to modulate cell growth behavior. By the addition and selection of the hydrophobic substituent, treatment methods are provided that allow for improved efficiency of the application of the therapeutic compounds of the invention to tissues, organs, and body parts. A change in the balance of the hydrophobic (lipophilic) and hydrophilic (ionic) properties of the cyclodextrin compounds of this invention, can favorably alter the biological impregnation of the membrane, with the consequence of improving the absorption within the blood plasma, by means of of the oral application. The compounds of the invention further alter interaction and retention by different biochemical components of cells, tissues, organs and body parts, such as with, or in fatty components, lipids, and other lipophilic materials. The treatment methods can be adapted in this way to the specific needs of the specific type and pathology involved. The compounds of the present invention conveniently possess the pharmacological activity of modulating cell proliferation in many therapeutic applications. It is believed, for example, that the application of the compounds of this invention to cardiovascular pathologies is beneficial. It has been shown that heparin possesses beneficial lipolytic activities (see A.C. Asmal et al., Bri., J. Clin, Pharmacology 1, 531-533, 1979), but its anticoagulant activity avoids its use in sufficiently high doses. The new and improved "heparin mimics" of this invention, devoid of that anticoagulant activity, overcomes this problem. It has been noted that the proliferation of human soft muscle cells (smc) is promoted by lipoprotein (see D.J. Grainger et al., Science 260, 1655-1658, 1993). With regard to methods for inhibiting soft-tissue cell proliferation, inhibiting plaque elongation, or preventing restenosis after angioplasty, the present invention provides for the use of modified cyandexcide polyanionic compounds, preferably polysulfates. The compounds provide additional benefits by impaction in the plaque and other areas of cardiovascular injury, by optimal bonding to those plaque areas and plaque removal by lipolytic or dissolution mechanisms. The present invention, therefore, provides methods of treating cardiovascular disorders. These methods include the inhibition of restenosis after angioplasty and the reduction of blood lipids. Methods that use sufficient numbers and lengths of hydrophobic (lipophilic) substituents linked to the critically sulfated cyclodextrin or other strongly anionic DC are especially useful for these purposes. The "one-sided" anionic cyclodextrins are particularly preferred for use in the methods of the invention. It has already been demonstrated that the ordinary beta-CD cavity is capable of reducing cholesterol in mammals (see M. Riottot et al., Lipids 28, 3, 181-188, 1993). However, it is well known that cyclodextrins have limited solubility of a few milligrams per milliliter, and are hemolytic at a concentration of almost 5 milligrams / milliliter. On the other hand, the addition of the anionic groups to the cyclodextrin molecule removes this hemolytic activity (see P.B. Weisz et al., Biochem. Pharm. 45, 1011-1016, 1993). The present invention also provides methods for inhibiting the proliferation of endothelial cells, the generation of endothelium, and the generation of new vascular systems, capillaries and blood vessels. These processes are known as neovascularization. This is related, therefore, with methods of prevention or inhibition of neovascularization or angiogenesis. The therapeutic use of the compounds of this invention provides improved effectiveness of those treatments by optimizing absorption or direction, as discussed above. The methods employ compositions comprising the polyanionic cyclodextrins of the present invention, together with an additional angiostatic agent of the type discussed previously. The present invention further provides improved antiangiogenic treatment methods, by applying the antiangiogenic compound in a more intimate manner than by an independent compound in a composition. In one embodiment, the angiostatic compound, being hydrophobic, such as an angiostatic steroid, is covalently linked to the polyanionic cyclodextrin, as the hydrophobic substituent, or as one of the hydrophobic substituents of the compound of this invention. In another embodiment, the hydrophobic or partially hydrophobic angiostatic molecule is combined with the "one-sided" polyanionic cyclodextrin to form the inclusion complex of the present invention. As noted above, it has been proven that the random binding of the sulfate groups in the cyclodextrin molecule inconveniently eliminates the internalization of the molecules by inclusion complexation with the cyclodextrin. The present invention gives attention to this problem by providing the anionic "one-sided" cyclodextrins, such as sulphated, which leaves an open door, so to speak, for internalization (inclusion complexation). Both embodiments provide uniformly dosed portions of the anionic cyclodextrin of the present invention, and the angiostat of interest. This is particularly useful in the preparation and use of these angiostatic compounds, such as steroids which have a very limited solubility in water. The compounds of this invention now unexpectedly make possible the solubilization of hydrophobic therapeutic agents, by inclusion complexation. Another aspect of the invention provides methods for treatment in the area of oncology. Although we do not wish to be bound by any particular theory, it is believed that treatment with the composition of the present invention inhibits the creation of new capillaries necessary for the growth of tumors. This results in the tumor having an insufficient supply of essential nutrients. Thus, when tumors in mammals, including humans, are treated in accordance with the present invention, they do not grow and may even die. Among the tumors contemplated as responsive to the composition and methods of this invention are Cellular Reticulum Sarcoma, Lewis Lung Carcinoma, Melanoma B-16, and Bladder Carcinoma, as well as others. It appears that neither mature blood vessels, not growing, nor mature vascular tissue, not growing, are affected by treatment with the compositions of the present invention. Inhibition of angiogenesis, in accordance with the present invention, in addition to its effect on tumor regression and metastasis in animals having a tumor, may be effective in the treatment of many ailments, as described hereinafter . The present invention further provides a method for the treatment of a number of non-tumor disorders which are characterized by pathological cells or tissue growth, including angiogenesis. In this manner, the invention provides a method for the treatment of mammals, including humans, afflicted with a number of non-neoplastic pathological conditions including rheumatoid arthritis, in which the proliferation of abnormal capillary growth appears in newborns and may persist for up to 2 years; angiofibromas which develop in the nasopharynx; psoriasis, in which excessive proliferation and fall may depend on abnormal capillary growth in the dermis. Additionally, the present invention provides a method for the treatment of a number of ophthalmic pathologies which are associated with undesirable angiogenesis and neovascular glaucoma. It is a further purpose of this invention to provide methods for treatments in which it is desirable to promote rather than inhibit, cell growth, cell proliferation, or repair or production of tissue or blood vessel. In this way, the compounds of the present invention can also positively modulate cell proliferation. This is the case in wound healing and repair to tissue and body parts due to either injury or surgical procedures, as well as deterioration or loss of structures due to pathologies. In all these cases, it is desired to have methods to apply, and preferably to optimally direct, protein growth factors to the place of the desired repairs. For these purposes, the polyanionic cyclodextrins of this invention are combined with a suitable protein growth factor to form a composition suitable for wound healing. The polyanionic CD, when contacted with a fluid medium containing the growth factor protein, for example, (HBGF), for a sufficient amount of time, produces an electrostatic (external) complex. Said complex is suitable as the composition to be applied to the site of the injury. It is significant to note that it is also known that electrostatic complexation, as in the case of heparin, stabilizes the protein molecule against enzymatic degradation that could result otherwise. Therefore, the application of the complex will not only provide a convenient method of applying a therapeutic agent directly to the cell and tissue, but will also conveniently improve the half-life and bioavailability of the growth factor protein applied as a complex. electrostatic protected. In the United States Patents of North American Numbers 5,019,562 and 5,183,809, the patents of which are incorporated herein by reference, describe in detail the doses of the compounds or compositions of the present invention, as well as the methods of their administration. It is considered within the skill of the art to routinely determine the appropriate amounts and mode of administration of the compounds of the invention. 7. METHODS FOR MAKING THE COMPOUNDS OF THE INVENTION In order to arrive at the CD compounds of the present invention having at least ten anionic compounds, for example, sulfate groups per molecule, the method of the invention uses the CD 2- positions, and 3-, since there are only seven 6- positions available.Also, for the "one-sided" anion substitution of the critical density of the anion of ten or more anions, such synthesis must result in all or almost all additions of anion to the side 2,3- In addition, the addition in that number on only the side 2,3- requires the use of the two positions 2- and 3- for the addition of the anion, since there are only seven of each available The discussion and examples are based on the use of beta-cyclodextrin It is noted that the limitation of the ten required anions will not change these restrictions in the addition methods for alpha-cyclodextrin, where There are six of each of positions 2-, 3, 6-, or for the gamma-cyclodextrin, where there are eight of each. Meanwhile, for this invention, the use of beta-cyclodextrin, particularly beta-cyclodextrin tetradecasulfate, over the use of alpha- or gamma-cyclodextrin is preferred, the discussion of this form of CD should not be considered as limiting this invention only to the beta form of the cyclodextrin. Alpha- and CD-gamma are also adaptable and are considered suitable for practice in the present invention, particularly in such forms as will adequately serve the purpose of one skilled in the art. To achieve critical sulfation as well as the addition of substantially hydrophobic substituents, a general method is provided as follows. The compounds of the present invention can be synthesized under conditions that allow hydrophobic substitution at the 2-, 3 or 6- positions of the cyclodextrin. The only limitation is that the critical minimum number of anionic groups of at least about 10 are provided in these positions. The preferred anionic group is sulfate. You can also choose to add the hydrophobic chain, one that does not include any reaction center that reacts with the sulfating agent. The product, after purification if necessary, can be reacted with a sulfating agent at a sufficient severity (concentration, time, temperature) to achieve sulfation of at least about ten units of secondary hydroxyl. The sulfating agent is typically one of the following: the sulfur trioxide complex of pyridine, chlorosulfonic acid, trimethylamine sulfur trioxide complex. Sulfation is achieved in a non-protic solvent, such as dimethylformamide, pyridine, or dimethylsulfoxide.

In order to obtain a strictly "one-sided" polyanionic CD product, and since the primary hydroxyl group in the 6-position of the sugar units is generally more reactive, someone can react and eventually replace a desired hydrophobic compound only in these hydroxyl groups under sufficiently mild reaction conditions so that the secondary hydroxyl groups (positions 2-, and 3-) remain unreacted. Also, for these hydrophobic substituents, one chooses a hydrophobic chain product that will not include any reaction center that reacts with the sulfating agent that is subsequently used. The so-called CD product substituted is reacted, after purification, with a sulfating agent at a severity (concentration, time, temperature) sufficient to achieve a minimum critical sulfation of at least about ten of the secondary hydroxyl units. The sulfating agent and the solvent are as described above. The following examples are provided to illustrate this invention. However, they should not be considered in any way as limiting the scope of the invention. 8. EXAMPLES EXAMPLE 1. Cyclodextrin with more than 10 sulfate groups and thioethyl group substitutions as hydrophobic components. Compound A. This is represented in Figure 3, where R = -S-CH2-CH3. The heptakis polysulfate compound (6-ethansulfide-6-deoxy) -beta-cyclodextrin was produced as follows: 8.64 grams of dry beta-cyclodextrin was added to a stirred solution of 42 grams of triphenylphosphine (TPhP) and 8.2 milliliters of bromine. in 160 milliliters of dimethylformamide (DMF). The mixture was stirred for hours at 83 ° C, concentrated by evaporation to one third of its volume, adjusted to pH 9.5 by the addition of 3M NaOMe in methanol (65 milliliters), with external cooling, stirred at room temperature. environment for 1 hour to decompose the esters, and emptied into ice water. The precipitate was washed in water, redissolved in dimethylformamide, again precipitated in methanol and washed in 3 x 1.5 liters of methanol to yield the product of heptakis (6-ethanesulfuro-6-deoxy) -beta-cyclodextrin (Int. 1) 1.45 grams of dry sodium hydride in 100 milliliters of dimethylformamide was added, stirred at 0 ° C under nitrogen at 4.8 milliliters of ethanetiol. To this solution, after stirring for 30 minutes, a solution of 2.4 grams of Int. 1 in 35 milliliters of dimethylformamide was added. at 0 ° C for 7 hours. After 40 hours at room temperature the unreacted ethanol was removed, reduced to 1/4 of its volume and precipitated in 600 milliliters of water. The white precipitate of heptakis (6-ethanesulfuro-6-deoxy) -beta-cyclodextrin was washed twice with methanol (Int.2). 3.2 grams of sulfur trioxide pyridine complex was added to a solution of 0.7 gram of Int. 2 in 35 milliliters of dry pyridine, and reacted at 25 ° C for 3 days. The pyridine was evaporated to precipitate a solid, and 30 milliliters of a 10 percent sodium acetate solution was added. The mixture was stirred for one hour, then methanol (250 milliliters) was added to form a suspension. The methanol was evaporated widely on a rotary evaporator. 250 milliliters of ethanol produced a precipitate, said precipitate was allowed to settle, and then washed in an additional 100 milliliters of ethanol. The solid was passed through chromatography on Sephadex with water as the levigant. A fraction of 0.7 grams of melting point of ca. 205-210 ° C, given an elemental sulfur analysis of 21.77 weight percent, which corresponds to the product heptakis polysulfate (6-ethanesulfuro-6-deoxy) -beta-cyclodextrin with approximately 12 sulfate groups, corresponding to the compound of Figure 3, with R = -S-CH2-CH3 bonded to the 6-position primary C-atom, thereby producing a four-atom hydrophobic chain element. This compound is also a "one-sided" polyanionic cyclodextrin.

EXAMPLE 2. A cyclodextrin with more than 10 sulfate groups and substitutions of the thiooctyl group as hydrophobic components. Compound B. This compound is represented by Figure 3, with R being -S- (CH) 7-CH 3 -. Heptakis polysulfate compound was produced (6-O-octansulfide-6-deoxy) -beta-cyclodextrin as follows: To a solution of 0.62 grams of dry sodium hydride in 100 milliliters of dimethylformamide, stirred at 0 ° C under nitrogen, were added, dropwise, 4.1 grams of loctanetiol. To this solution, after stirring for 30 minutes, 3.14 grams of Int. 1 were added gradually, stirred at room temperature for 50 hours and drained in 1 liter of methanol. The yellow precipitate was collected by filtration, washed in water (2 x 300 milliliters) and in methanol (2 x 200 milliliters) and dried under vacuum, yielding 3.8 grams of heptakis (6-octane sulfide-6-deoxy) -beta- cyclodextrin. For crystallization, 0.12 grams were dissolved in dimethylformamide and dilute ethanol, producing after two days colorless crystals with a melting point of 238-240 ° C. To a solution of 2.03 grams of Int. 3 in 700 milliliters of pyridine, 6.36 grams of sulfur trioxide pyridine complex was added and stirred at 100 ° C for 18 hours. The pyridine was evaporated to a precipitated solid, which was dissolved in 60 milliliters of a sodium acetate solution and stirred for one hour. This solution was added to methanol (500 milliliters). The precipitate was washed in ethanol (2 x 20 milliliters). The precipitate yielded 5.4 grams of crude product containing heptakis polysulfate (6-octane sulfide-6-deoxy) -beta-cyclodextrin. A fraction, after passing through Sephadex 25 chromatography, produced a product with a sulfur analysis of 16.64 5 which indicated the presence of approximately 11 sulfate groups. This corresponds to the compound of Figure 3, with R = -S- (CH 2) 7 -CH 3 bonded to the primary C-atom of position 6, thereby producing a hydrophobic chain element of ten atoms. This compound is also an "one-sided" anionic polyanionic CD. EXAMPLE 3. Cyclodextrin with more than 10 sulfate groups and substitutions of the thiophenyl group as hydrophobic components. Compound C. This compound is represented by Figure 4. The heptakis polysulfate compound (6-pentanesulfuro-6-deoxy) -beta-cyclodextrin was produced in a manner strictly analogous to example 5, taking the pentylthiol the place of ethylthiol and octylthiol of examples 4 and 5, respectively. The product of this reaction corresponds to the compound of Figure 3, with R = -S- (CH 2) 4 -CH 3 bonded to the secondary C atom of the 6-position, thereby producing a hydrophobic seven-atom chain element. This compound is also an "one-sided" anionic polyanionic CD. Examples 1, 2, and 3 illustrate the compounds of this invention with varying amounts of hydrophobicity achieved by variable length substituents of non-polar bonding groups of atoms. It is possible to have an incorporation of side chains with only portions of them containing chains of groups of non-polar atoms. It would be expected that said compounds would have the hydrophobicity expressed in that portion of the environment, that is, at a molecular scale, although the entire molecule may exert less average hydrophobicity in tests such as the solubility of the entire molecule. An example is the structure of D in Figure 4, which was obtained in a multi-step synthesis described in Example 6. EXAMPLE 4. Heptakis polysulfate [6-hepta-0- (4-carboxyphenyloxy) octanoate)] - beta-cyclodextrin. Compound D, Figure 4. This compound has more than 10 sulfate groups and complex substituents on its side chain, containing a 12-member chain of non-polar atoms, but terminated by a polar group. 8-bromooctanoate (Int. 4) was produced from dicyclohexylcarbodiimide and bromooctanoic acid, in 4- (dimethylamino) pyridine. It was independently produced, ali-hydroxybenzoate (Int. 5) from DBU, allyl bromide and 4-hydroxybenzoic acid, in acetonitrile. Then the benzyl 8- [4- (allyloxycarbonyl) phenyloxy] octanoate (Int. 6) was produced by reduction (Int. 5) in dimethylformamide and subsequently reacting the product with (Int. 4) in dry dimethylformamide. A solution of (Int. 6) at 0 ° C in dichloromethane with pyrrolidine in the presence of triphenylphosphine and tetrakis (triphenylphosphine) palladium to produce benzyl 8- (4-carboxyphenyloxy) octanoate (Int. 7). Β-Cyclodextrin was independently reacted with tert-butyldimethylsilane chloride in pyridine to produce heptakis (6-0-tert-butyldimethylsilyl) -beta-cyclodextrin (Int. 8). This product was reacted with benzyl bromide and sodium hydride in anhydrous dimethylformamide to yield heptaquis (2,3-tetradecabenzyl-6-0-hepta-tert-butyldimethylsilyl) -beta-cyclodextrin (Int. 9). Benzylation of all hydroxy groups 2- and 3-was achieved by reaction with benzyl bromide and sodium hydride in dimethylformamide, followed by removal of the silyl protecting groups in THF, washing, drying, and concentrated the organic phase yielding heptaquis (2, 3-0-tetradecabenzyl) -beta-cyclodextrin (Int. 10). The condensation reaction of Int. 10 with Int. 7 was performed using the classical DCC-DMAP treatment for more than 30 hours, followed by purification on silica gel, and hydrogenation in ethanol on Pd / activated charcoal, provided heptaquis [-0] (4-carboxyphenyloxy octanoate)] - beta-cyclodextrin (Int. 11). This final intermediate was sulfated by the standard sulfation procedure to produce the product of Figure 4D. Elemental analysis indicated that sulfation of approximately 14 sulfate groups was achieved. The product showed inhibitory activity on rat soft muscle cell proliferation by the thymidine uptake assay in vi tro. The variation of the homopolar property contributions of the products made by the choice of the composition of the substituents attached to the 6-position of the critically sulfated CD was determined, requiring ten or more sulphate groups that occupied the sites of the 2- position. and 3- of the sugar units, by the comparative solubilities of said compounds as compared to the cyclodextrin polysulfide of the prior art which lacked hydrophobic substitutions. EXAMPLE 5. The aqueous solubilities of the beta-cyclodextrins observed were as follows: Number of hydrophobic solubility compound (grams / 100 ml) chain groups 27 ° C 0 ° C CD 0 ca. 1 less than 1.0 CDS 0 ca. 127 ca. 95 A 4 66 52 C 7 ca. 58 25 B 10 11.3 8.2 CD is beta-cyclodextrin without sulfation CDS is cyclodextrin polysulfate of the prior art It is noted that by providing a previously determined length of a hydrophobic substitute chain, a desired reduction in solubility can be obtained. In this case, agents of solubilities greater than about 20 to 30 grams / milliliters (measured at 0 ° C) are achieved by chain lengths less than about seven; therefore solubilities less than about 20 grams / 100 milliliters are obtained with chain lengths greater than seven or eight. Of course, additional elongations of said chains can be achieved routinely. Very low solubilities may be achieved with chain lengths of 20 or more units of hydrophobic atom, providing estimated solubilities below about 0.2 grams / 100 milliliters or less. For negligible solubilities, one can synthesize oligomers or polymers containing said units as taught in this invention. EXAMPLE 6 It was important to ensure that the addition of a hydrophobic group to the cyclodextrins would not interfere with or destroy the cellular biological activity of the CDS. The above compound was tested with the greatest number of hydrophobic (lipophilic) groups in the molecule namely compound B, which had octyl groups, and indeed as many as seven such chains, to see its activity to inhibit growth (proliferation ) of soft muscle cells (smc). The test is the one used by some of us previously (see H.C. Herrmann et al., Arteriosclerosis and Thrombosis 13, 924, 1993). The following example is briefly described, along with the resulting observations. EXAMPLE 7. Human umbilical vein soft muscle cells were allowed to join microtiter well plates 96 coated with fibronectin, incubated for 72 hours with varying concentrations of the CD test sample and 10 percent fetal calf serum. . The cells were fixed, stained with blue-black naphthol, lysis was caused, and quantified by light absorption at 530 nm. ID50 is the concentration of the test sample that results in the maximum average inhibition of proliferation.

Sample ID50 (mg / ml) CDS 1.0 ± 0.2 B ca. 0.1 to 1.0 It was evident that the highly substituted sample was at least as active as, with strong evidence for considerably higher activity, than the cyclodextrin polysulfate of the prior art. It should be noted that a smaller ID50 corresponds to correspondingly lower dose requirements to achieve the same inhibitory effect. It should also be noted that the total number of hydrophobic (lipophilic) atomic groups placed in sample B was actually approaching 8 times (C chain) 7 (number of substituents added at position -6) or a total of fifty-six, or if we count the S atom and the secondary C atoms in the complete chain, we are dealing with a total of 60 non-polar groups that have been added without interfering, and, in fact, increasing the biological activity of the cyclodextrin polysulfide. The invention provides "one-sided" polyanionic CDs that have a critical number of anionic substituents. These "single-sided" CDs admit host molecules freely within the internal cavity for complexation. This provides the resulting multiple potential for storage and pharmacological application of the host molecules. The following examples will illustrate said ability of the compounds of this invention, in contrast to the CDS composition of the prior art. EXAMPLE 8. When the water phase containing compound A was contacted with toluene and subsequently examined to see the absorption of ultraviolet rays in the ultraviolet absorption band by the bne ring, the molar amount of toluene found approached that of the content of the cyclodextrin-compound A. The same result was observed when the compound C was similarly tested. In contrast, the repetition of the unsubstituted CDS test of the prior art showed no evidence of ultraviolet rays of toluene associated with that compound. EXAMPLE 9. 2.3 mM of the "single-sided" cyclodextrin sample of Example 4 was contacted with a suspension of 2.9 mM of a steroid, 19-norandrost-4-ene3, 17-dione. A change in adsorption maximum of 244 to 240 nm was observed, indicating formation of the inclusion complex. No such change could be observed with the ordinary CDS of the prior art. The following example describes the generation of a polysulfonated cyclodextrin with hydrophobic substituents (lipophilic) or substituents that have pharmacological activity themselves. It is about a method for producing a bound hydrocortisone-cyclodextrin polysulfate, thus combining the two activities required for the anti-angiogenesis activity: a mimic of heparin and an angiostat. EXAMPLE 10. The preparation of a beta-cyclodextrin hydrocortisone linked covalently with the treatment of the cyclodextrin with tosyl chloride in pyridine was initiated to obtain the monotosylate in the 6- position of the cyclodextrin (2 in Figure 5). The displacement of the tosylate group with potassium iodide at 95 ° C for 3 hours in the dark compound produced 3 in Figure 5. Independently, the hydrocortisone was protected (5S-pregnane-3oi, 17a, 20-triol, 11, 20 dione) with 2, 2-dimethoxypropane in dimethylformamide, at 60 ° C, to obtain 5β-pregnane-3a-ol, 17Oi, 20-acetonide, 11,20 dione) (compound 4 in Figure 5). The side chain was prepared by protecting 4- (4-aminophenyl) butyric acid with a Z group to give 4- (4-carbobloxy-aminophenyl) butyric acid (5 in Figure 5). Condensation was achieved between compound 4 and 5 in dimethylformamide at room temperature, in the presence of DCC and DMAP for 19 hours, and the product purified by chromatography on silica gel to provide compound 6 in 67 percent yield, followed by hydrogenation over palladium on activated carbon to provide 1-hydrocortisone-4- (4-aminophenyl) butyric acid ester (compound 7 in Figure 5). The covalent bond of compound 4 with compound 7 was achieved by reaction in dimethylformamide at 60 ° C for 48 hours to obtain the hydrocortison-cyclodextrin linkage, compound 8. This compound is subjected to sulfation as shown in the last step of Figure 5. Similar and other steroids can be linked to the cyclodextrin structure. Variations, improvements and simplifications are possible in the synthetic steps and can be obtained by the person skilled in the technique of organic synthesis. It should be understood that the foregoing detailed description and the accompanying examples are merely illustrative and should not be construed as limitations on the scope of the invention, which is defined exclusively by the appended claims and their equivalents. Various changes and modifications may be made, including without limitation, those relating to the substituents, derivatives, syntheses, formulations and / or methods of use of the invention, without departing from the spirit and scope of the present invention.

Claims (57)

1. A substituted polyanionic cyclodextrin compound having cell growth modulation properties, wherein at least ten substituents per molecule of cyclodextrin are anions of a strong acid selected from the group consisting of sulfate, nitrate, sulfonate or phosphate, the compound being associated with a physiologically acceptable cation, and at least one additional substituent being a hydrophobic compound of a linker chain of at least three carbons in length.

2. The substituted polyanionic cyclodextrin compound of Claim 1, wherein the hydrophobic substituent is selected from the group consisting of alkyl, aryl, ester, ether, thioester and thioether.

3. The substituted polyanionic cyclodextrin compound of Claim 1, wherein the number and length of the hydrophobic substituent is of a size sufficient to reduce the water solubility of the cyclodextrin compound by at least about thirty percent.

4. The substituted polyanionic cyclodextrin compound of Claim 3, wherein the number and length of the hydrophobic substituent is of a size sufficient to substantially increase the absorption of the cyclodextrin compound into the bloodstream of a mammal, after administration oral.

5. The substituted polyanionic cyclodextrin compound of Claim 1, wherein at least one of the hydrophobic substituents is pharmacologically active.

6. The substituted polyanionic cyclodextrin compound of Claim 5, wherein the pharmacologically active substituent has angiostatic, antiviral or antibiotic activity.

7. The substituted polyanionic cyclodextrin compound of Claim 6, wherein the pharmacologically active substituent is an angiostat.

8. The substituted polyanionic cyclodextrin compound of Claim 7, wherein the angiostat is a steroid.

9. A "one-sided" substituted polyanionic cyclodextrin compound having cell growth modulation properties, wherein at least ten substituents per molecule of cyclodextrin are anions of a strong acid selected from the group consisting of sulfate, nitrate, sulfonate or phosphate, the compound being associated with a physiologically acceptable cation, the anions being located substantially only in the 2- and 3- positions of the cyclodextrin, and at least one additional substituent being a hydrophobic compound of a linking chain of at least three carbons in length, hydrophobic substituent which is located substantially only in the 6-positions of the cyclodextrin.

The "one-sided" substituted polyanionic cyclodextrin compound of claim 9, wherein the hydrophobic substituent is selected from the group consisting of alkyl, aryl, ester, ether, thioester and thioether.

The substituted polyanionic cyclodextrin compound of Claim 10, wherein the number and length of the hydrophobic substituent is of a size sufficient to reduce the water solubility of the cyclodextrin compound by at least about thirty percent.

The substituted polyanionic cyclodextrin compound of Claim 10, wherein the number and length of the hydrophobic substituent is of a size sufficient to substantially increase the absorption of the cyclodextrin compound within the bloodstream of a mammal, after administration oral.

13. The substituted polyanionic cyclodextrin compound of Claim 9, wherein at least one of the hydrophobic substituents is pharmacologically active.

14. The substituted polyanionic cyclodextrin compound of Claim 13, wherein the pharmacologically active substituent has angiostatic, antiviral or antibiotic activity.

15. The substituted polyanionic cyclodextrin compound of Claim 14, wherein the pharmacologically active substituent is an angiostat.

16. The substituted polyanionic cyclodextrin compound of Claim 15, wherein the angiostat is a steroid.

17. An inclusion complex of a "one-sided" substituted polyanionic cyclodextrin of Claim 9, and a host molecule having a hydrophobic structure capable of penetrating into and complexing with the cyclodextrin.

The inclusion complex of a "one-sided" substituted polyanionic cyclodextrin of Claim 17, wherein the host molecule is a pharmacologically active compound of limited solubility in water.

19. The inclusion complex of Claim 18, wherein the host molecule is selected from the group consisting of a steroid, vitamin A, beta-carotene, or structurally related compounds.

20. A method for modulating the growth behavior of living cells, comprising contacting the cells with a substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per molecule of Cyclodextrin are anions of strong acids selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, and one or more additional substituents comprise hydrophobic elements of a linking chain of at least three carbons in length .

21. A method as in Claim 21, wherein the living cells are soft muscle cells.

22. A method as in Claim 20, wherein the contact comprises the substituted polyanionic cyclodextrin and at least other pharmacologically active compound.

23. A method as in Claim 22, wherein the living cells are endothelial cells.

24. A method as in Claim 22, wherein the added active compound is one of the group of steroids, vitamin A, beta-carotene, or structurally related compounds.

25. A method for modulating the growth behavior of living cells, comprising contacting the cells with a "one-sided" substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per molecule of cyclodextrin are strong acid anions selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, the anions being located substantially only in positions 2- and 3 - of the sugar units, and one or more additional substituents comprise hydrophobic elements of a linking chain of at least three carbons in length, which substituents are substantially located only in the 6- positions of the sugar units.

26. An improved drug application method to a mammal, comprising the steps of contacting (1) a "one-sided" substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per molecule of cyclodextrin are anions of strong acids selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, the anions being located substantially only in the 2- and 3- positions of the sugar units, and one or more additional substituents comprise hydrophobic elements of a linking chain of at least three carbons in length and whose substituents are located substantially only in the 6- positions of the sugar units with (2) a pharmacologically molecule active that has a structure sufficiently hydrophobic to be able to p Entering inside and ending with the hydrophobic internal cavity of the cyclodextrin molecule, for a period of time sufficient to create an inclusion complex of (2) in (1), and apply the inclusion complex to the mammal.

27. An improved drug application method to a mammal, according to Claim 26, wherein the method of application to the mammal is by intravenous injection of the inclusion complex contained in a physiologically acceptable solvent.

28. An improved drug application method to a mammal, according to Claim 26, wherein the method of application to the mammal is by injection into the sinuvial fluid of the inclusion complex contained in a physiologically acceptable solvent.

29. An improved drug application method to a mammal, according to Claim 26, wherein the method of application to the mammal is by topical application of the inclusion complex contained in a physiologically acceptable medium, to a surface of cellular tissue of the mammal.

30. A method of therapeutic intervention in pathologies caused by soft tissue cell proliferation at a cell tissue site, wrapped in this condition a physiologically acceptable composition comprising a substituted polyanionic cyclodextrin compound having cell growth modulation properties, in wherein at least about ten substituents per cyclodextrin molecule are strong acid anions selected from the group of sulfate, nitrate, sulfonate or phosphate, the compound being associated with a physiologically acceptable cation, and one or more additional substituents comprising hydrophobic elements of a link chain of at least three carbons in length.

31. A method as in Claim 30, wherein the pathology to be treated is the prevention of restenosis after angioplasty in a blood vessel.

32. A method of therapeutic intervention in cardiovascular pathologies caused or accompanied by hypercholesterolemia in a mammal, including a human, which comprises applying to the cardiovascular system a "one-sided" substituted polyanionic cyclodextrin compound having cell growth modulation properties, wherein at least about ten substituents per molecule of cyclodextrin are strong acid anions selected from the group of sulfate, nitrate, sulfonate or phosphate, the compound being associated with a physiologically acceptable cation, the anions being located substantially only at the 2-position. - and 3- of the sugar units of the cyclodextrin.

A method as in Claim 32, wherein the polyanionic cyclodextrin is a "one-sided" substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per molecule of cyclodextrin are anions of strong acids selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, the anions being located substantially only in the 2- and 3- positions of the sugar units, and one or more additional substituents comprise hydrophobic elements of a linking chain of at least three carbons in length, and whose substituents are located substantially only in the 6-positions of the sugar units.

34. A method of assisting in the wound healing process, which comprises applying to the cellular material surrounding the injured or damaged tissue or organ, including bone, a physiologically acceptable carrier comprising (1) a substituted polyanionic cyclodextrin compound that has pronounced cell growth modulation properties, wherein at least about ten substituents per cyclodextrin molecule are strong acid anions selected from the sulfate group, nitrate, sulfone-to or phosphate, associated with a physiologically acceptable cation, and (2) a growth factor protein.

35. A method of assisting in the wound healing process, comprising applying to the cellular material surrounding the injured or damaged tissue or organ, including bone, a physiologically acceptable carrier comprising (1) a substituted polyanionic cyclodextrin compound that has pronounced cell growth modulation properties, wherein at least about ten substituents per cyclodextrin molecule are strong acid anions selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, and one or more additional substituents comprising hydrophobic elements with a linker chain of at least three carbons, and (2) a growth factor protein.

36. A method of assisting in the healing process of lesions, as in Claim 32, wherein (1) is a "one-sided" substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per molecule of cyclodextrin are anions of strong acids selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, the anions being located substantially only at the 2- positions and 3- of the sugar units, and one or more additional substituents comprise hydrophobic elements of a linking chain of at least three carbons in length, and whose substituents are located substantially only in the 6- positions of the sugar units, and (2) a growth factor protein.

37. A method of assisting in the process of transplantation of cellular material such as cells, organelles, tissue, or organs, in order to assist in the acceptance of cellular material by the host environment, which comprises contacting said cellular material before and / or after implantation, with a physiologically acceptable composition comprising (1) a substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per molecule of cyclodextrin are selected strong acid anions from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, and one or more additional substituents comprising hydrophobic elements with a linking chain of at least three carbons, and (2) a protein of growth factor .

38. A method of assisting in the process of transplantation of cellular material such as cells, organelles, tissue, or organs, in order to assist in the acceptance of cellular material by the host environment, which comprises contacting said cellular material before and / or after implantation, with a physiologically acceptable composition comprising (1) a "one-sided" substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per cyclodextrin molecule are anions of strong acids selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, said anions being located substantially only in the 2- and 3- positions of the sugar units, and one or more additional substituents comprise hydrophobic elements of a linking chain of at least three carbons of length, and whose substituents are located substantially only in positions 6- of the sugar units, and (2) a growth factor protein.

39. A method of aid for the revascularization of the vasculature in the condition of ischemia, which comprises contacting the environment of the tissue involved in ischemia, with a physiologically acceptable composition comprising (1) a substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per cyclodextrin molecule are strong acid anions selected from the sulfate group , nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, and one or more additional substituents comprising hydrophobic elements with a linker chain of at least three carbons, and (2) a growth factor protein.

40. A method of aid for revascularization of the vasculature in the condition of ischemia, comprising contacting the environment of the tissue involved in ischemia, with a physiologically acceptable composition comprising (1) a substituted polyanionic cyclodextrin compound "of a only side "having pronounced cell growth modulation properties, wherein at least about ten substituents per cyclodextrin molecule are strong acid anions selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically cation acceptable, said anions being located substantially only in the 2- and 3- positions of the sugar units, and one or more additional substituents comprise hydrophobic elements of a linking chain of at least three carbons in length, and whose substituents are substantially localized only in positions 6- of the units of sugar, and (2) a growth factor protein.

41. A method of inhibiting the biological process of neovascularization comprising contacting the wrapped tissue, with a physiologically acceptable composition comprising (1) a substituted polyanionic cyclodextrin compound having pronounced cell growth modulation properties, wherein at least about ten substituents per molecule of cyclodextrin are anions of strong acids selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, and one or more additional substituents comprising hydrophobic elements with a linking chain of at least three carbons, and (2) an angiostatic compound.

42. A method of inhibiting the biological process of neovascularization, as in claim 41, wherein the angiostatic compound is one selected from the group of an angiostatic steroid, an angiostatic protein, an angiostatic derivative of the Fumagillin family, an angiostatic derivative of retinoic acid.

43. A method of inhibiting the biological process of neovascularization comprising contacting the wrapped tissue with a physiologically acceptable composition comprising (1) a "one-sided" substituted polyanionic cyclodextrin compound having cell growth modulating properties pronounced, wherein at least about ten substituents per cyclodextrin molecule are strong acid anions selected from the group of sulfate, nitrate, sulfonate or phosphate, associated with a physiologically acceptable cation, said anions being located substantially only at the 2- positions and 3- of the sugar units, and one or more additional substituents comprise hydrophobic elements of a linking chain of at least three carbons in length, and whose substituents are located substantially only in the 6-positions of the sugar units, and (2) is an angiostatic compound.

44. A method of inhibiting the biological process of neovascularization, as in claim 43, wherein the angiostatic compound is one selected from the group of an angiostatic steroid, an angiostatic protein, an angiostatic derivative of the Fumagillin family, an angiostatic derivative of retinoic acid.

45. A method of inhibiting the biological process of neovascularization comprising contacting the wrapped tissue, with a physiologically acceptable composition comprising a "one-sided" anionic cyclodextrin complex, as in Claim 17, wherein the host molecule is an angiostatic compound selected from the group of an angiostatic steroid, an angiostatic protein, an angiostatic derivative of the Fumagillin family, an angiostatic derivative of retinoic acid.

46. A method of inhibiting the biological process of neovascularization, according to claim 45, for the therapeutic purpose of inhibiting the growth of tumors, wherein the contact is made by injecting said composition into a growing tumor or its immediate tissue environment.

47. A method of inhibiting the biological process of neovascularization, according to claim 45, for the therapeutic purpose of inhibiting the pathological growth of blood capillaries, inside the cornea of a mammal, where contact is made by topical application of the composition in an aqueous formulation of eye drops applied topically to the eye.

48. A method of inhibiting the biological process of neovascularization, according to claim 45, for the therapeutic purpose of inhibiting the pathological growth of blood capillaries, inside the skin of a mammal, as a result of psoriasis or other angiogenic dermatological disease. , wherein the contact is made by the topical application of said composition in an aqueous or otherwise physiologically acceptable medium to the skin.

49. A method of inhibiting the biological process of neovascularization, according to Claim 45, for the purpose of the therapeutic treatment of an angiostatic disease, wherein the contact is made by intravenous injection of the composition into the circulating bloodstream of the mammal to be treated.

50. A method of producing a sulfated inclusion complex of a fully or partially hydrophobic compound as the host, and a polyanionic cyclodextrin as the host, comprising the steps of 1) forming an inclusion complex by contacting the polyanionic cyclodextrin with the compound to be included, and then 2) sulfatar the inclusion complex obtained in step 1.

51. The method of Claim 50, wherein the host compound is a steroid.

52. An inclusion complex obtained by the method of Claim 50.

53. The inclusion complex of the Claim 52, wherein the included compound is a steroid.

54. The inclusion complex of the Claim 53, wherein the steroid is hydrocortisone.

55. The inclusion complex of the Claim 52, wherein the compound is an antiviral.

56. The inclusion complex of Claim 55, wherein the antiviral is AZT.

57. The inclusion complex of Claim 52, wherein the included compound is a retinoid.