MXPA96006716A - Soluble rapamycin steres in a - Google Patents

Abstract

DU composed of the structure in which R1 and R2 are each independently hydrogen or -COCH2-S-CH2CH2-O-CH2- (CH2OCH2) n -CH2-O-CH2CH2-OCH3; and n = 8-450: with the proviso that R1 and R2 are not both hydrogen, which compound is useful as an immunosuppressant, anti-inflammatory, antifungal, antiproliferative and antitumor agent

Description

STERES DE RAPMCG-fflft fíOT.TmT.Kfi ftffl? Ft This invention relates to methoxypoly (ethylene glycol) water-soluble rapamycin esters, and to a method for using them to induce immunosuppression, and in the treatment of transplant rejection, graft-versus-host disease, autoimmune diseases, inflammation diseases, leukemia. T-cell lymphoma, solid tumors, fungal infections and hyperproliferative vascular disorders. Rapamycin is a macrocyclic trieno antibiotic produced by Streptomyces hisroscopicus. which is found to have antifungal activity, particularly against Candida albicans, both in vi-tro and in vivo [C. Vezinaet al., J. Antibiot. 28, 721 (1975); S.N. Sehgal et al., J. Antibiot. 28, 727 (1975); H. Baker A. et al., J. ntibiot. 31,539 (1978); U.S. Patent Number 3,929,992; and North American Patent Number 3,993,749]. Rapamycin alone (American Patent Number 4,885,171) or in combination with picibanil (Patent North American Number 4,401,653) has been shown to have antitumor activity. R. Martel et al., [Can. J. Physiol. REF: 23693 Pharmacol. 55, 48 (1977)] describes that rapamycin is effective in the model of experimental allergic encephalomyelitis, a model for multiple sclerosis; in the adjuvant arthritis model, a model for rheumatoid arthritis; and effectively inhibits the formation of IgE-like antibodies. The immune-suppressive effects of rapamycin have been described in FASEB 3, 3411 (1989). Cyclosporin A and FK-506, other macrocyclic molecules, have also been shown to be effective as immunosuppressive agents, therefore, they are useful in avoiding rejection of transplants [FASEB 3, 3411 (1989); FASEB 3, 5256 (1989); R. Y. Calne et al., Lancet 1183 (1978); and North American Patent Number ,100,899]. It has also been shown that rapamycin is useful for preventing or treating systemic lupus erythematosus [Patent American Number 5,078,999], pulmonary inflammation [U.S. Patent Number 5,080,899], insulin dependent diabetes mellitus [Fifth Int. Conf. Inflamm. Res. Assoc. 121 (Abstract), (1990)], proliferation of smooth muscle cells and thickening of the intima after vascular damage [Morris, R.J. Heart Lung Transplant 11 (pt.2): 197 (1992)], leukem, ia / lympholas of adult T cells [European Patent Application 525,960 Al], and ocular inflammation [European Patent Application 532,862 Al] It has been demonstrated that the monoacylated and diacylated derivatives of rapamycin (esterified in the positions 28 and 43) are useful as antifungal agents (U.S. Patent Number 4,316,885) and are used to make water soluble aminoacyl prodrugs of rapamycin (U.S. Patent Number 4,650,803). Recently, the numbering convention of rapamycin has been changed; therefore, according to the Chemical Abstracts nomenclature, the esters described above would be in positions 31 and 42. U.S. Patent No. 5,023,263 describes the preparation and use of 42-oxorapamycin and U.S. Patent Number 5,023,264 describes the preparation and use of the 27-oximes of rapamycin. Polyethylene glycol (PEG) is a neutral, linear or branched polymer, available in a variety of molecular weights and is soluble in water and in most organic solvents. At molecular weights less than 1000 are colorless and viscous liquids; the highest molecular weight PEGs are waxy and white solids. The melting point of the solid is proportional to the molecular weight, reaching a plateau at 67 ° C. Molecular weights range from a few hundred to approximately 20,000 and are commonly used in biological and biotechnological applications. Of great interest in the biomedical areas is the fact that PEG is non-toxic and has been approved by the FDA for internal consumption.

DBSCRIPTION OF Li This invention provides methoxypoly-ethylene glycol) rapamycin esters having the structure wherein R1 and R2 are each independently, hydrogen or -COCH2-S-CH2CH2-0-CH2- (CH: OCH2) n-CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen.

The compounds of this invention are water soluble prodrugs of rapamycin which are useful as suppressant, anti-inflammatory, antifungal, antiproliferative and antitumor agents. Of the compounds of this invention, it is preferred that n = 8-200; it is further preferred that n = 8-135. The most preferred mes are those in which n = 8-20, and those in which n = 90-120. The compounds of this invention which are esterified at positions 42 or 31, 42, can be prepared by initially acylating positions 42 or 31 and 42 of rapamycin with an acylating agent having the general structure X-CH2C02H, where X is a suitable leaving group such as iodine, in the presence of a coupling reagent , such as dicyclohexylcarbodiimide (DCC) and a base such as dimethylaminopyridine (DMAP) to provide the acylated rapamycin at position 42 or 31, 42, having the following structure: The mixtures of the esters 42 and 31, 42 can be separated by chromatography. The reaction of rapamycin acylated with monomethylpoly (ethylene glycol) thiol in the presence of a base such as PROTON SPONGE ([1, 8- (bis (dimethylamino) -naphthalene, N, N, N ', N' -tetramethyl-l-8 -naphthalenediamine]) or sodium bicarbonate provides the desired esters 42 or 31, 32 of this invention.The 31-esters of this invention can be prepared by protecting the 42 -alcohol of rapamycin with a protecting group, such as a tert-butyldimethylsilyl group, followed by esterification of position 31 by the procedures described above The preparation of the 42-silylethers of rapamycin is described in US Patent Nu Bl 5,120,842, which is incorporated herein by reference. Removal of the protecting group provides the compounds esterified in position 31. In the case of the tert-butyldimethylsilyl protecting group, the deprotection can be carried out under moderately acidic conditions, such as acetic acid / water / THF. The method of deprotection is described in Example 15 of U.S. Patent No. 5,118,678 which is incorporated herein by reference. Having the position 31 esterified and the position 42 deprotected, the position 42 can be esterified using a different acylating agent that reacts with the alcohol in the position 31 to provide compounds having different esters in positions 31 and 42. Alternately , the compounds esterified at position 42, are prepared as described above, can be reacted with different acylating agents to provide compounds having different esters at positions 31 and 42. This invention. it also encompasses analogous esters of other rapamycins such as, but not limited to, 29-demethoxyrapamycin [U.S. Patent Nu 4,375,464, 32-demethoxyrapamycin under the nomenclature of C.A.]; rapamycin derivatives in which double bonds at positions 1, 3 and / or 5 have been reduced [U.S. Patent Nu 5,023,262]; 29-desmethylrapamycin [U.S. Patent No. 5,093,339, 32-desmethylrapamycin under the nomenclature of C.A.]; 7,29-bisdesmethylrapamycin [U.S. Patent Nu 5,093,338, 7, 32-desmethylrapamycin under the nomenclature of C.A.]; 27-hydroxyrapamycin [U.S. Patent Nu 5,256,790] and 15-hydroxyrapamycin [U.S. Patent Nu 5,102,876]. This invention also encompasses esters at position 31 of 42-oxorapamycin [U.S. Patent Nu 5,023,263]. The descriptions of the North American Patents mentioned in the foregoing are incorporated herein by reference. The reagents used in the preparation of the compounds of this invention can be obtained commercially or they can be prepared by conventional or standard procedures described in the literature. The immunosuppressive activity for the representative compounds of this invention is established in a standard pharmacological test procedure of skin graft in pinch-ja that measures the immunosuppressive activity of the compound tested as well as the ability of the compound tested to inhibit or treat the rejection of transplants. The procedure for this standard pharmacological test procedure and the results obtained are given in the following. The representative compounds of this invention are evaluated by a live test procedure designed to determine the survival time of a pinched skin graft from male BALB / c donors to female C3H (H-2K) receptors. The method is adapted from Billingham R.E. and Medawar P.B., J. Exp. Biol. 28: 385-402, (1951). Briefly, a donor graft is pinched on the back of the recipient as an allograft, and an isograft is used as a control in the same region. The receptors are treated with varying concentrations of test compounds intraperitoneally or orally. Rapamycin is used as a test control. Untreated receivers serve as rejection control. The graft is checked daily and observations are recorded until the graft dries and forms a black scab. This is considered as the day of rejection. The mean graft survival time (number of days ± SD) of the treatment group is compared with the control group. The results are expressed as the average survival time, in days. Untreated skin grafts (control) are usually rejected in the following 6-7 days. A survival time of 11.67 ± 0.63 for rapamycin at 4 mg / kg i.p. Since the compounds of the invention are prodrugs of rapamycin, the doses provided in the following are provided in equivalent doses of rapamycin (6.2 mg of the compound of Example 2 contains the equivalent of 1 mg of rapamycin). The results obtained for the compound of Example 2, PEG-5000, and untreated control are given in the following table.

Survival Time Compound Dosage * I & (Average t D.E.) Example 2 20 mg / kg p.o. 12.50 ± 0.22 Example 2 5 mg / kg p.o. 11.33 + 0.33 Example 2 1.25 mg / kg p.o. 8.67 ± 0.21 Example 2 4 mg / kg i.p. 12.67 ± 0.21 Example 2 1 mg / kg i.p. 11.33 ± 0.21 Example 2 0.25 mg / kg i.p. 9.5 ± 0.22 Control 7.00 ± 0.00 PEG-5000 7.00 ± 0.00 * The dose of the compound of Example 2 is provided in equivalent doses of rapamycin. The results of this standard pharmacological test procedure demonstrate immunosuppressive activity for the compounds of this invention. Additionally, the results obtained in the skin graft test procedure demonstrate the ability of the compounds of this invention to treat or inhibit transplant rejection. Based on the results of these standard pharmacological test procedures, the compounds are useful in the treatment or inhibition of transplant rejection such as kidney grafts, heart, liver, lung, bone marrow, pancreas (islet cells), cornea, small intestine and cutaneous allografts as well as heart valve grafts; in the treatment or inhibition of graft-versus-host disease; in the treatment or inhibition of immune diseases such as lupus, rheumatoid arthritis, diabetes ellitus, myasthenia gravis, and multiple sclerosis and inflammation diseases such as psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease, lung inflammation (including asthma, chronic obstructive pulmonary disease, emphysema, acute respiratory distress syndrome, bronchitis and the like) and ocular uveitis. Due to the profile of activity obtained, the compounds of the invention are also considered to have anti tumor, anti-fungal and antiproliferative activities. Therefore, the compounds of this invention are useful for treating solid tumors including sarcomas and carcinomas, such as astrocytomas, prostate cancer, breast cancer, small lung cancer and ovarian cancer, leukemia / adult T-cell lymphoma; Mycotic infections and hyperproliferative vascular diseases such as restenosis and atherosclerosis. When used for restenosis, it is preferred that the compounds of this invention be used to treat restenosis that occurs after the angioplasty procedure. When used for this purpose, the compounds of this invention may be administered prior to the procedure, during the procedure, post-procedure or in any combination of the foregoing. When administered for the treatment or inhibition of previous disease states, the compounds of this invention can be administered to a mammal per day oral, parenteral, intranasal, intrabronchial, transdéprica, topical, intravaginal or rectal. The compounds of this invention are particularly advantageous as immunosuppressive, anti-inflammatory, antifungal, antiproliferative and antitumor agents due to their water solubility. For example, rapamycin has a solubility of 1.2 μg / ml in water, while the compound of Example 2 has a solubility of >100 mg / ml, which facilitates simplicity in terms of formulation and administration. It is contemplated that when the compounds of the invention are used as an immunosuppressant or anti-inflammatory agent, they may be administered together with one or more additional immunoregulatory agents. Such additional immunoregulatory agents include, but are not limited to, azatropiopine, corticosteroids, such as prednisone and methylprednisolone, cyclophosphamide, rapamycin, cyclosporin A, FK-506, OKT-3 and ATG. By combining the compounds of this invention with other medicaments or agents to induce immunosuppression or to treat inflammatory conditions, amounts of each of the agents are required to obtain the desired effect. The basis for such combination therapy is established by Stepkowski whose results show that the use of a combination of rapamycin and cyclosporin A at therapeutic dose significantly prolongs the survival time of a cardiac allograft [Transplantation Proc. 23: 507 (1991)]. The compounds of this invention can be formulated in a pure manner or with a pharmaceutical carrier to a mammal in need thereof. The pharmaceutical carrier can be solid or liquid. A solid carrier can include one or more substances which also act as flavoring agents, lubricants, solubilizers, suspension improving agents, fillers, fluidizing agents, compression aids, binders or tablet disintegrating agents; It can also be an encapsulating material. In powders, the carrier is a finely divided solid which is mixed with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary compression properties in suitable proportions and the desired shape and size are shared. The tablets and powders preferably contain up to 99% of the active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidine, low melting point waxes, and ion exchange resins. .

Liquid carriers are used to prepare solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both, or pharmaceutically acceptable oils or fats. The liquid carrier may contain other suitable pharmaceutical additives such as solubilizers, buffers, preservatives, sweeteners, flavoring agents, suspension improving agents, thickening agents, colors, viscosity regulators, stabilizers and osmo-regulators. Suitable examples of liquid carriers for oral and parenteral administration include water (which partially contains additives as in the above, for example cellulose derivatives, preferably a solution of sodium carboxymethylcellulose), alcohols (including monohydric alcohols and polyhydric alcohols, example glycols) and its derivatives, lecithins and oils (for example fractionated coconut oil and peanut oil). For parenteral administration, the carrier can be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are useful in sterile liquid form compositions for parenteral administration. The liquid carrier for pressurized compositions may be a halogenated hydrocarbon or other pharmaceutically acceptable propellant. Liquid pharmaceutical compositions which are sterile solutions or suspensions may be used by, for example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be given intravenously. The compound can also be administered orally either in the form of a liquid or solid composition. The compounds of this invention can be administered rectally in the form of a conventional suppository. For administration by inhalation or intranasal or intrabronchial insufflation, the compounds of this invention can be formulated in aqueous or partially aqueous solution, which can then be used in the form of an aerosol. The compounds of this invention can also be administered transdermally by the use of a transdermal patch containing the active compound and a carrier that is inert to the active compound, which is not toxic to the skin and which allows the release of the agent for systemic absorption in the bloodstream via the skin. The carrier can take any of the various forms such as creams and ointments, pastes, gels and occlusion devices. Creams and ointments may be viscous liquid or semi-solid emulsions of the oil-in-water or water-in-oil type. Pastes consisting of absorptive powders dispersed in petroleum or hydrophilic petroleum containing the active ingredient may also be suitable. A variety of occlusion devices may be used to release the active ingredient into the bloodstream such as a semipermeable membrane that covers a reservoir containing the active ingredient with or without a carrier, or a matrix containing the active ingredient. Other occlusion devices are known in the literature. In addition, the compounds of this invention can be used as a solution, cream or lotion by formulation with pharmaceutically acceptable carriers containing 0.1-5 percent, preferably 2% of the active compound, which can be administered in an area affected by mushrooms. Dosage requirements vary with the particular compositions used, the route of administration, the severity of the symptoms presented and the particular subject in question. Based on the results obtained in the standard pharmacological test procedures, the projected daily dosages of active compound would be 0.1 μg / kg 100 mg / kg, preferably between 0.001-25 mg / kg, and more preferably between 0.01. - 5 mg / kg. Treatment will usually start with small dosages less than the optimal dose of the compound. Subsequently, the dosage is increased until the optimum effect is reached: under the circumstances; precise dosages for oral, parenteral, nasal or ír administration. rabronquial will be determined by the doctor who makes the administration based on the experience with the individual subject treated. Preferably, the pharmaceutical composition is in unit dosage form, for example, as tablets or capsules. In such form, the composition is subdivided into unit doses containing the appropriate quantities of the active ingredient; the unit dosage forms may be packaged compositions, for example packets, vials, ampules, pre-filled syringes or sacks containing liquids. The unit dosage form can be, for example, a capsule or tablet itself, or it can be an appropriate amount of such compositions in package form. The following examples illustrate the preparation and biological activities of the representative compounds of this invention.

Example 1 Ester di 42- p oacetato cift rapap-icina Rapamycin (0.5 g, 5.5 x 10"4 moles) and DMAP (3.0 mg) are dissolved in 15 ml of anhydrous methylene chloride, iodoacetic acid (0.123 g, 6.6 x 10" 4 moles) and DCC (0.136 g, 6.6 x 104 moles) in 20 ml of anhydrous methylene chloride and the mixture is transferred to a dropping funnel; this mixture is added slowly to the rapamycin solution for a period of 30 minutes, with agitation. The reaction mixture is stirred at room temperature for an additional hour. The resulting solution is filtered through a sintered glass filter. The filtrate is transferred to a separatory funnel and washed first with two 40 ml portions of a sodium bicarbonate solution (5.5 g / 100 ml) and then with water (2 x 50 ml). The methylene chloride layer is dried with 3 g of anhydrous sodium sulfate for 5 hours. Subsequently, the sodium sulfate is removed by filtration and the methylene chloride is evaporated, which gives 0.53 g of a pale yellow solid compound. CLAP shows the presence of 42-ester (55%), 31-ester (9.2%), 31, 42-diester (17%) and unreacted rapamycin (17%).

The pure 42-iodoacetate is isolated by preparative CLAP on a Primesphere column (250 x 50 mm, 10 microns). Rapamycin 42-iodoacetate elutes at 8.1 minutes with the use of 80% methylene chloride (solution A) and 20% solution B. Solution B consists of 85% methylene chloride and 15% solution C (2 : 1 = methanol: isopropanol). The eluate is evaporated and the residue is treated in methylene chloride, dried and evaporated, which gives 0.206 g of a solid compound. (+) Ionic MS m / z 1099.5 (M * NH4K; (-) ionic MS m / z 1080.5. "H NMR (400 MHz, DMSO-d6) ß 3.78 (s, 2H, CO-CH2-I), 4.54 (m, 1H, H-42) Analysis calculated for C53H80NO4I: C, 58.83; H, 7. Four. Five; N, 1.29. Found: C, 58.97; H, 7.64; N, 1.36.

Ejßpplo 2 42-áatar flfl «CU" "^" * «" m-at-rreipc-li 1.wngl nnl) t-Ác > ? 5QQ0 Method 1 The rapamycin 42-iodoacetate ester (0.1 g, 9.2 x 10"5 mol) is dissolved in methylene chloride (15 mL) and methanol (15 mL), then mPEG-SH 5000 is added. (0.6 g, 1.2 x 10'4 moles) and PROTON SPONGE (20 mg, 9.3 x 105 moles) to this reaction solution, which is stirred at room temperature overnight. Then, 10 mg of PROTON SPONGE are again added and the reaction solution is again stirred at room temperature overnight.

The reaction is suspended by adding ether (200 ml). The white precipitate is filtered and washed with ether (3 x 20 ml), which gives 0.59 g of the crude or untreated product. The crude product is further purified by preparative CLAP on a Zorbax C8 column (250 x 20 mm) by using a gradient of solution A with 30-80% solution B. Solution A consists of 90% 0.1% TEAA ( tetraethylammonium acetate), pH 4.5, buffered and 10% acetonitrile. Solution B consists of 10% TEAA at 10%, buffered pH 4.5 and 90% acetonitrile. The ester of 42-mPEG-S 5000 rapamycin acetate elutes at 21 minutes. The aqueous phase is extracted with methylene chloride (2 x 50 mL). The organic layer is dried with anhydrous sodium sulfate for 14 hours, and concentrated to a volume of 10 ml under reduced pressure. The product precipitates upon addition of 100 ml of ether. The white precipitate is collected on a sintered glass filter and washed with ether (3 x 20 ml), which gives 109.6 mg of the product.

Method 2 The rapamycin 42-iodoacetate ester is dissolved (0.5 g 4.6 x 10 ~ 4 moles), 130 ml of 50% acetonitrile-containing solution and 50% aqueous NaHCO3 solution (0.1 M) are dissolved. The solution is purged with N2 for 10 minutes. In order to verify the initial reactive condition, samples of 20 μl are extracted and added to 1 ml of acetonitrile. The solution is filtered in 10 μl of the sample undergoing analysis by CLAP. MPEG-SH 5000 (3.15 g) is added, 6.3 x 10"4 moles) to the reaction solution over a period of 1.5 h, and the reaction is stirred at room temperature for another 1.5 hrs. Another 20 μl is removed from the sample, mixed with 1 ral of acetonitrile, They are filtered and injected into the CLAP system The results of the CLAP analysis show that the rapamycin iodoacetate is quantitatively converted to rapamycin 42-MPEG S 5000 acetate ester.The reaction mixture is extracted with methylene chloride (2 x 500 ml) After the organic layer is dried with anhydrous NasS04 and filtered, the filtrate is concentrated to a volume of about 20 ml.The final crude product is precipitated by adding 250 ml of ether; this suspension is then filtered and dried under vacuum, which provides 3.13 g of dry white material. Unreacted mPEG-SH is removed by preparative CLAP as described in method 1. EM (MADI / TOF shows a primary molecular weight of 5877.47 for the product and 4923.66 for initial mPEG-SH 5000. The difference in mass (953.81) exactly matches the rapamycin 42-acetate moiety (953.6) The ester side chain can be represented by the formula -COCH2-S-CH2CH2-0-CH2- (CH.OCH2) n-CH2-0- CH2CH2-OCH3, where n is an average of 108 repetitive units XH NMR (400 MHz, CDCl3) d 2.84 (t, '2H, S-CH2 £ H2), 3.27 (s, 2H, CO- £ H2- S), 3.36 (s, 3H, -0 £ H3), 3.64 (m, 4H,? - £ H2-C02-O), 4.69 (m, 1H, H-42) .MS (MALDI / TOF) m / z 5877.47 (average molecular weight) UV (CH3CN)? max 268, 278, 290 nm.

Example 3 31. 32-diiodo --- rapamicitm catato Rapamycins (0.5 g, 5.5. X 10"4 moles), DCC (0.28 g, 1.4 x 10" 3 moles) and DMAP (30 mg) are dissolved in anhydrous methylene chloride (15 ml). Iodoacetic acid (0.25 g, 1.4 x 10"3 moles) is added to the reaction solution, and the reaction mixture is stirred for one hour at room temperature, then the solution is filtered through a sintered glass filter. The filtrate is washed with two portions of a solution of sodium bicarbonate (5.5 g / 100 ml) and with water (2 x 50 ml), the methylene chloride layer is dried with 3 g of anhydrous sodium sulfate for 5 hours. The sodium sulfate is then removed by filtration and the methylene chloride is evaporated, yielding 0.63 g of a pale yellow solid material The CLAP data indicates that 99.4% of rapamycin 31,42-diiodoacetate has been formed. (+) Ionic MS m / z 1272.3 (M + Na) *. X H RN (400 MHz, CDCl 3) d 3.77 (c, 2 H, C 0- £ H2-I, 31-ester), 3,784 (s, 2 H, CO -fiHj-I, 42-ester), 4.31 (d, 1H, H-31), 4.54 (m, 1H, H-4.2).

Ejespío 4 tiQl 5Q0Q It dissolves rapamycin 31, 42-diiodoacetate (5.99 mg, 4.8 x 10"s moles) in 70 ml of a 50% solution of CH3CN-50% NaHCO3 (0.1 M) .The solution is purged with nitrogen for 10 minutes.MPEG is added to the reaction solution -SH 5000 (0.778 g, 1.56 x 10"4 moles). After the reaction, the solution is stirred for 30 minutes, a 30 μl sample is extracted, mixed with 1 ml of acetonitrile and filtered. The sample (10 μl) is subjected to analysis by CLAP. The data indicate that rapamycin iodoacetate is 100% converted to rapamycin ester 31, 42 -di (mPEG-S-5000 acetate). The reaction mixture is extracted with dry methylene chloride (2 x 300 mL). The methylene chloride layer is dried with anhydrous sodium sulfate and filtered. The filtrate is concentrated to a volume of 20 ml. The product is precipitated by adding 250 ml of ether, filtered and dried under vacuum, which gives 0.22 g of a white material. MS (MALDI / TOF) shows an average molecular weight of 10983.6 Da. The ester side chains can be represented by the formula -COCH2-S-CH2CH2-0-CH2- (CH2OCH2) n-CH2-0-CH2CH2-OCH3, wherein n is an average of 108 repetitive units. 'H NMR (400 MHz, CDC13) d 3.23 (c, 2H, C0- £ H2-S, 31-ester), 3.25 (s, 2H, CO- £ H2-S, 42-ester), 4.65 (m, 1H, H-42), 5.25 (d, 1H, H-31).

Ejespío 5 Lpoli (atilenglicol) thiol 750 The rapamycin 42-iodoacetate ester (100 mg, 9.2 x 10"5 moles) is dissolved in 30 ml of a 50% solution of CH3CN-50% NaHCO3 (0.1 M) .The solution is purged with nitrogen for 10 minutes mPEG-SH-750 (1.25 g, 1.67 x 10"3 moles) is added to the reaction solution. After the reaction solution is stirred for 30 minutes, 30 μl of the sample is extracted, added with 1 ml of CH 3 CN and filtered. The sample (10 μl) is subjected to analysis by CLAP. The data indicates that rapamycin 42-iodoacetate is converted quantitatively to the 42-mPEG-S-750 acetate ester of rapamycin. The reaction mixture is extracted with dry methylene chloride (2 x 300 mL). The methylene chloride layer is dried with anhydrous sodium sulfate and filtered. The filtrate is concentrated to a volume of 20 ml. The product is precipitated by adding 250 ml of ether, filtered and dried under vacuum, which provides 80 g of a viscous oil liquid material. The side chain ester can be represented by the formula -COCH2-S-CH2CH2-0-CH: - (CH; 0CH2) n-CH2-0-CH2CH2-0CH3, where n is an average of 14 repetitive units.

ESI-MS (M + NH4) * m / z 1460.1 (n = 10), 1548.1 (n = 12), 1592.2 (n = 13), 1636.2 (n = 14), 1680.1 (n = 15), 1724. 0 (n = 16), 1769.0 (n = 17), 1812.9 (n = 18); (M + NH4) 2 * m / z 871.3 (n = 16), 893.5 (n = 17), 915.5 (n = 18), 937.0 (n = 19), 959.4 (n = 20), 981.4 (n = 21) ). It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects to which it relates. Having described the invention as above, property is claimed as contained in the following:

Claims (16)

1. A composite of the structure characterized in that R1 and R2 are each independently hydrogen or -C0CH2-S-CH2CH2-0-CH2- (CH20CH2) n -CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen.
2. 2. The compound according to claim 1, characterized in that n = 8-200.
3. 3. The compound according to claim 1, characterized in that n = 8-135.
4. 4. The compound according to claim 1, characterized in that n = 8-20.
5. 5. The compound according to claim 1, characterized in that n = 90-120.
6. 6. The compound according to claim 1, characterized in that it is the 31,42-diester of methoxypoly (ethylene glycol) thiol 5000.
7. 7. The compound according to claim 1, characterized in that it is the 42-ester with methoxypoly (ethylene glycol) thiol 5000.
8. 8. The compound according to claim 1, characterized in that it is the 42-ester with methoxypoly (ethylene glycol) thiol 750.
9. 9. A method for treating rejection of transplants or graft-versus-host disease in a mammal in need thereof, characterized in that it comprises administering to the mammal an anti-rejection effective amount of a compound of the structure wherein R1 and R2 are each independently hydrogen or -COCH2-S-CH2CH2-0-CH2- (CH2OCH2) n-CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen.
10. 10. A method for treating a fungal infection in a mammal in need thereof, characterized in that it comprises administering to the mammal an antimycotic effective amount of a compound of the structure wherein R1 and R2 are each independently hydrogen or -COCH2-S-CH2CH2-0-CH3- (CH2OCH2) n-CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen.
11. 11. A method for treating rheumatoid arthritis of a mammal in need thereof, characterized in that it comprises administering to the mammal an antiarthritic effective amount of a compound of the structure wherein R1 and R2 are each independently hydrogen or -COCH2-S-CH2CH2-0-CH2- (CH2OCH2) n-CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen.
12. 12. A method for treating restenosis in a mammal in need thereof, characterized in that it comprises administering to the mammal an antiproliferative effective amount of a compound of the structure wherein R1 and R2 are each independently hydrogen or -COCH2-S-CH2CH2-0-CH2- (CH2OCH2) n-CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen.
13. 13. A method for treating lung inflammation in a mammal in need thereof, characterized in that it comprises administering to the mammal an anti-inflammatory effective amount of a compound of the structure wherein R1 and R2 are each independently hydrogen or -COCH2-S-CH2CH2-0-CH2- (CH2OCH2) n-CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen.
14. 14. A method for treating solid tumors in a mammal in need thereof, characterized in that it comprises administering to the mammal an antitumor effective amount of a compound of the structure wherein R1 and R2 are each independently hydrogen or -COCH2-S-CH2CH2-0-CH2- (CH2OCH2) n-CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen.
15. 15. The method according to claim 14, characterized in that the solid tumor is a carcinoma or sarcoma.
16. 16. A pharmaceutical composition, characterized in that it comprises a compound of the structure wherein R1 and R2 are each independently hydrogen or -COCH2-S-CH2CH2-0-CH2- < CH2OCH2) n-CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen, and a pharmaceutic carrier. A composite of the structure wherein R1 and R2 are each independently hydrogen or -C0CH2-S-CH2CH2-0-CH2- (CH20CH2) n -CH2-0-CH2CH2-OCH3; and n = 8 - 450; with the proviso that R1 and R2 are not both, hydrogen, compound which is useful as an immunosuppressive, anti-inflammatory, antimyotic, antiproliferative and antitumor agent.