IE913302A1

IE913302A1 - Aminoesters of rapamycin

Info

Publication number: IE913302A1
Application number: IE330291A
Authority: IE
Original assignee: American Home Prod
Priority date: 1990-09-19
Filing date: 1991-09-19
Publication date: 1992-02-25
Also published as: AU8659991A; FI931203A; EP0549727A1; AU653175B2; HUT65763A; MX9101139A; HU9300776D0; JPH06501012A; WO1992005179A1; FI931203A0; PT98990A

Abstract

A compound of structure (I), wherein R<1>, R<2>, and R<3> are each, independently, hydrogen, or R<4>; R<4> is (a), (b), or (c); R<5> is hydrogen, alkyl, aralkyl, -(CH2)qCO2R<8>, -(CH2)rNR<9>CO2R<10>, carbamylalkyl, aminoalkyl, hydroxyalkyl, guanylalkyl, mercaptoalkyl, alkylthioalkyl, indolylmethyl, hydroxypehnylmethyl, imidazoylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl, alkoxy, hydroxy, cyano, halo, nitro, carbalkoxy, trifluoromethyl, amino, or a carboxylic acid; R<6> and R<9> are each, independently, hydrogen, alkyl, or aralkyl; R<7>, R<8>, and R<10> are each, independently, alkyl, aralkyl, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri-substituted; R<11> and R<12> are each, independently, alkyl, aralkyl, or phenyl which is optionally mono-, di-, or tri-substituted; X is (d), O, or S; R<13? and R?14> are each, independently, hydrogen or alkyl; Y is CH or N; m is 0-4; n is 0-4; p is 1-2; q is 0-4; r is 0-4; t is 0-4; u is 0-4; wherein R<5>, R<6>, m, and n are independent in each of (e) subunits when p=2; or a pharmaceutically acceptable salt thereof, with the proviso that R<1>, R<2>, and R<3> are not all hydrogen, further provided that R<1>, R<2> and R<3> are not all (a), and still further provided that t and u are not both 0 when X is O or S, which by virtue of its immuno-suppressive activity is useful in treating transplantation rejection, host vs. graft disease, autoimmune diseases, and diseases of inflammation, and by virtue of its antifungal activity is useful in treating fungal infections.

Description

BACKGROUND OF THE INVENTION This invention relates to novel esters of rapamycin and a method for using them in the treatment of transplantation rejection, host vs. graft disease, autoimmune diseases, diseases of inflammation, and fungal infections.

Rapamycin is a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus. which was found to have antifungal activity, particularly against Candida albicans, both in vitro and in vivo [C. Vezina et al., J. Antibiot. 28, 721 (1975); S.N. Seghal et al., J. Antibiot. 28, 727 (1975); H. A. Baker et al., J. Antibiot. 31, 539 (1978); U.S. Patent 3,929,992; and U.S. Patent 3,993,749], Rapamycin alone (U.S. Patent 4,885,171) or in combination with picibanil (U.S. Patent 4,401,653) has been shown to have antitumor activity. R. Martel et al.

[Can. J. Physiol. Pharmacol. 55, 48 (1977)] disclosed that rapamycin is effective in the experimental allergic encephalomyelitis model, a model for multiple sclerosis; in the adjuvant arthritis model, a model for rheumatoid arthritis; and effectively inhibited the formation of IgE-like antibodies.

The immunosuppressive effects of rapamycin have been disclosed in FASEB 3, 3411 (1989), rapamycin has been shown to be effective in inhibiting transplant rejection (U.S. Patent Application Ser. No. 362,544 filed June 6, 1989). Cyclosporin A and FK-506, other macrocyclic molecules, also have been shown to be effective as immunosuppressive agents, therefore useful in preventing transplant rejection [FASEB 3, 3411 (1989); FASEB 3, 5256 (1989); and R. Y. Caine et al., Lancet 1183 (1978)].

Mono- and diacylated derivatives of rapamycin (esterified at the 28 and 43 positions) have been shown to be useful as antifungal agents (U.S. Patent 4,316,885) and used to make water soluble prodrugs of rapamycin (U.S. Patent 4,650,803). Recently, the numbering convention for rapamycin has been changed; therefore according to Chemical Abstracts nomenclature, the esters described above would be at the 31-and 42-positions.

AHP-9580/9675-1-N1 -2DESCRIPTION OF THE INVENTION This invention provides derivatives of rapamycin which are useful as immunosuppressive, anti-inflammatory, and antifungal agents having the structure wherein R1, R2· and R3 are each, independently, hydrogen, or R4; R4 is O II [C(CH2)mCH(CH2)nN]pCO2R7 , o 11 11 -C-(CH2)lX(CH2)uCO2R11 , or R5 R6 COoR12R5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, -(CH2)qCO2R8, -(CH2)rNR9CO2R10, carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazolylmethyl or phenyl which is optionally mono-, di, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R6 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms; AHP-958O/9675-1-N1 -3R7, R8, and R10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or trisubstituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R11 and R12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R13 Xis -C-, 0, orS; R14 R13 and R14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms; YisCHorN; m is 0 - 4; n is 0 - 4; p is 1 - 2; q is 0 - 4; r is 0 - 4; t is 0 - 4; u is 0 - 4; O II wherein R5, R6, m, and n are independent in each of the[C(CH2)mCH(CH2)nN] I I R5 R6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R1, R2· and R3 are not all hydrogen, further provided that R1, R2· and R3 are not all O II — [C(CH2)mCH(CH2)nN]pCO2R7 , and still further provided that t and u are not I I R5 R6 both 0 when X is O or S.

AHP-9580/9675-1-Nl -4o II Of the compounds when R4 is — [C(CH2)mCH(CH2)nN]pCO2R7 , ι ι R5 R6 preferred members are those in which m = 0, n = 0, and p=l;m = 0, n = 0, and p = 2; n = 0, and R5 is -(CH2)qCO2R8; m = 0, n = 0, and R5 is -(CH2)rNR9CO2R10; and m = 0, n = 0, and R5 is hydrogen. Preferred compounds also include those O members in which R4 is —C-(CH2)tX(CH2)uCO2R .

The pharmaceutically acceptable salts may be formed from inorganic cations such as 10 sodium, potassium, and the like; mono-, di-, and trialkyl amines of 1-6 carbon atoms, per alkyl group and mono-, di-, and trihydroxyalkyl amines of 1-6 carbon atoms per alkyl group; and organic acids such as acetic, lactic, citric, tartaric, succinic, maleic, malonic, gluconic, and the like. Preferred basic salts are formed from sodium cations and tris(hydroxymethyl)methylamine.

The compounds of this invention can be prepared by acylating rapamycin with an acylating agent having the general structures where Z is OH;in the presence of a coupling reagent, such as dicyclohexy1-carbodiimide . The compounds of this invention also can be prepared using an anhydride or mixed anhydride Qrf the above described carboxylic acid as the acylating species. Alternatively, the acylating species can be an acid halide, where X can be Cl, Br or I. The acylating groups used to prepare the compounds of this invention are commercially available or can be prepared by methods that are disclosed in the literature .

Where it is desired to prepare acyl derivatives having two or three different R4 groups then sequential acylation may be performed using appropriate acylating agents as defined above, if necessary isolating the AHP-9580/9675-1-Nl -5desired product by appropriate purification techniques In general the 42 -position is acylated first and such a monoacylated product may be isolated prior to the second acylation and so forth. Appropriate protecting groups may be used to block any position where acylation is not required.

Immunosuppressive activity was evaluated in an in vitro standard 5 pharmacological test procedure to measure lymphocyte proliferation (LAF) and in two in vivo standard pharmacological test procedures. The first in vivo procedure was a popliteal lymph node (PLN) test procedure which measured the effect of compounds of this invention on a mixed lymphocyte reaction and the second in vivo procedure evaluated the survival time of a pinch skin graft.

The comitogen-induced thymocyte proliferation procedure (LAF) was used as an in vitro measure of the immunosuppressive effects of representative compounds. Briefly, cells from the thymus of normal BALB/c mice are cultured for 72 hours with PHA and IL-1 and pulsed with tritiated thymidine during the last six hours. Cells are cultured with and without various concentrations of rapamycin, cyclosporin A, or test compound. Cells are harvested and incorporated; radioactivity is determined. Inhibition of lymphoproliferation is assessed in percent change in counts per minute from non-drug treated controls. The results are expressed by the following ratio, or as the percent inhibition of lymphoproliferation of 1 μΜ. 3H-control thymus cells - HLrapamycin-treated thymus cells 3H-control thymus cells - H3-test compound-treated cells A mixed lymphocyte reaction (MLR) occurs when lymphoid cells from genetically distinct animals are combined in tissue culture. Each stimulates the other to undergo blast transformation which results in increased DNA synthesis that can be quantified by the incorporation of tritiated thymidine. Since stimulating a MLR is a function of disparity at Major Histocompatibility antigens, an in vivo popliteal lymph node (PLN) test procedure closely correlates to host vs. graft disease. Briefly, irradiated spleen cells from BALB/c donors are injected into the right hind foot pad of recipient C3H mice. The drug is given daily, p.o. from Day 0 to Day 4. On Day 3 and Day 4, tritiated thymidine is given i.p., b.i.d. On Day 5, the hind popliteal lymph nodes are removed and dissolved, and radioactivity counted. The corresponding left PLN serves as the control for the PLN from the injected hind foot. Percent suppression is calculated using the non-drug treated animals as allogenic control.

Rapamycin at a dose of 6 mg/kg, p.o. gave 86% suppression, whereas cyclosporin A at the same dose gave 43% suppression. Results are expressed by the following ratio: AHP-9580/9675-1-N1 -63H-PLN cells control C3H mouse - 3H-PLN cells rapamvcin-treated C3H mouse 3H-PLN cells control C3H mouse - 3H-PLN cells test compound-treated C3H mouse The second in vivo test procedure is designed to determine the survival time of pinch skin graft from male DBA/2 donors transplanted to male BALB/c recipients. The method is adapted from Billingham R.E. and Medawar P.B., J. Exp. Biol. 28:385402, (1951). Briefly, a pinch skin graft from the donor is grafted on the dorsum of the recipient as a homograft, and an autograft is used as control in the same region. The recipients are treated with either varying concentrations of cyclosporin A as test control or the test compound, intraperitoneally. Untreated recipients serve as rejection control. The graft is monitored daily and observations are recorded until the graft becomes dry and forms a blackened scab. This is considered as the rejection day. The mean graft survival time (number of days ± S.D.) of the drug treatment group is compared with the control group.

The following table summarizes the results of representative compounds of this invention in these three standard test procedures.

TABLE 1 Compound LAF* (ratio) PLN* (ratio) Skin Graft (davs + SD) Example 1 1.8 0.61 12.0 ±_1.6 Example 2 0.33 0.62 11.5 + 0.6 Example 3 0.20 + 9.0 ± 0.9 Example 4 4.9 0.18 12.3 ±0.5 Example 5 0.006 + 8.8 ± 0.9 Example 6 5.4 0.33 11.5 ±3.5 Example 7 3% at ΙμΜ** + 7.7 ± 1.5 Example 8 0.03 0.41 + Example 9 0.96 1.34 10.3 ± 0.8 Example 10 2.0 0.96++ 12.7 ± 1.2 Example 11 0.004 + 10.5 ± 1.3 Example 12 19.8 -2.87 12.0 ±2.0 Example 13 22% at ΙμΜ** + 7.0 ±0.6 Example 14 0.37 + 8.2 ± 1.2 Example 15 0.9 0.69 10.7 ± 1.2 AHP-9580/9675-I-N1 -7- Compound TABLE LAF* (ratio) 1 (Continued) PLN* (ratio) Skin Graft (davs + SE Example 16 3.27 1.04## 12.7 ± 0.9 Example 17 0.56 1.68### 10.2 ± 1.7 Example 18 0.02 1.11## 8.0 ± 1.7 Example 19 0.01 0.48 8.0 ± 0.9 Example 20 0.97 0.70 9.3 ± 1.6 Example 21 0.22 -1.93 12.0 + 1.7 Example 22 0.22 0.41 10.2+1.2 Example 23 0.18 0.39 10.8 +.0.8 Example 24 0.00 0.09 7.8 +_1.7 Rapamycin 1.0 1.0 12.0 ± 1.7 * Calculation of ratios was described supra.

** Result expressed as percent inhibition of lymphoproliferation at 1 μΜ.

+ Not evaluated ++ Results obtained using cremophore/ethanol as a vechicle for administration.

Ratios of 0.33 and 1.07 were also obtained using carboxymethyl cellulose as a vehicle for administration.

## Results obtained using cremophore/ethanol as a vechicle for administration. Ratios of 0.20 and 1.08 also were obtained using carboxymethyl cellulose as a vehicle for administration.

### A ratio of 0.42 also was obtained for this compound.

The results of these standard pharmacological test procedures demonstrate immunosuppressive activity both in vitro and in vivo for the compounds of this invention. Positive ratios in the LAF and PLN test procedures indicate suppression of T cell proliferation. As a transplanted pinch skin grafts are typically rejected within 6-7 days without the use of an immunosuppressive agent, the increased survival time of the skin graft when treated with the compounds of this invention further demonstrates their utility as immunosuppressive agents. While it appears that the compound disclosed by Examples 12 and 21 may cause T cell proliferation in the PLN test procedure, it is believed a negative ratio in this test procedure coupled with an increased survival time observed in the skin graft test procedure indicates a proliferation of Tsuppressor cells, AHP-9580/9675-1-N1 -8which are implicated in suppressing the immune response, (see, I. Roitt et al. Immunology, C.V.Moseby Co. 1989, p 12.8-12.11).

Antifungal activity of the compounds of this invention was measured against 5 5 strains of Candida albicans using a plate test procedure for measurement of inhibition. The following represents the typical procedure used. Compound to be tested was placed on sterile dried 1/4 plate disks, and allowed to dry. Agar plates were seeded with fungi and allowed to solidify. The impregnated disks were placed on the seeded Agar surface and incubated for the time required for the particular culture. Results are 10 expressed in MIC (pg/ml) to inhibit growth. The results of this test procedure showed that the compounds of this invention have antifungal activity; however, it was surprising that the compounds of this invention were less active than the parent compound, rapamycin.

Table 2* Strain of Candida albicans Compound ATCC 10231 ATCC 38246 ATCC 38247 ATCC 38248 2669 Example 1 >0.4 >0.4 >0.4 >0.4 >0.4 Example 2 0.1 0.2 0.2 0.2 0.1 20 Example 3 0.4 >0.4 >0.4 >0.4 0.4 Example 4 0.1 0.4 0.1 0.1 0.2 Example 5 >0.4 >0.4 >0.4 >0.4 >0.4 Example 6 0.1 >0.4 0.2 0.4 >0.4 Example 7 + + + + + 25 Example 8 >0.4 >0.4 >0.4 >0.4 >0.4 Example 9 0.4 >0.4 0.4 >0.4 >0.4 Example 10 0.2 >0.4 0.2 0.4 0.4 Example 11 >0.4 >0.4 >0.4 >0.4 >0.4 Example 12 0.2 >0.4 0.1 0.2 0.4 30 Example 13 >0.4 >0.4 >0.4 >0.4 >0.4 Example 14 >0.4 >0.4 >0.4 >0.4 >0.4 Example 15 >0.4 0.4 >0.4 0.4 0.4 Example 16 0.2 0.1 0.4 0.1 0.1 Example 17 >0.4 0.2 >0.4 0.2 0.4 35 Example 18 0.4 >0.4 >0.4 >0.4 >0.4 Example 19 0.4 >0.4 0.4 >0.4 >0.4 AHP-9580/9675-1-N1 -9Table 2* (Continued) Strain of Candida albicans Compound ATCC 10231 ATCC 38246 ATCC 38247 ATCC 38248 3669 Example 20 0.1 0.4 0.1 0.1 0.2 Example 21 0.4 >0.4 0.4 >0.4 >0.4 Example 22 0.2 >0.4 0.2 0.4 >0.4 Example 23 0.1 >0.4 0.2 0.4 >0.4 Example 24 >0.4 >0.4 >0.4 >0.4 >0.4 Rapamycin 0.003 0.025 0.003 0.006 0.025 * expressed as MIC (gg/ml) + not evaluated Based on the results of these standard pharmacological test procedures, the compounds are useful in the treatment of transplantation rejection such as, heart, kidney, liver, bone marrow, and skin transplants; autoimmune diseases such as, lupus, rheumatoid arthritis, diabetes mellitus, myasthenia gravis, and multiple sclerosis; and diseases of inflammation such as, psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease; and fungal infections.

The compounds may be administered neat or with a pharmaceutical carrier to a mammal in need thereof. The pharmaceutical carrier may be solid or liquid.

A solid carrier can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, 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 in admixture 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 compacted in the shape and size desired. The powders and tablets 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, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.

Liquid carriers are used in preparing 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 AHP-9580/9675-1-N1 - 10can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators. Suitable examples of liquid carriers for oral and parenteral administration include water (partially containing additives as above, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and their derivatives, and oils (e.g. fractionated coconut oil and arachis oil). For parenteral administration, the carrier can also 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 can be halogenated hydrocarbon or other pharmaceutically acceptable propellent.

Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by, for example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously. The compound can also be administered orally either in liquid or solid composition form.

Preferably, the pharmaceutical composition is in unit dosage form, e.g. as tablets or capsules. In such form, the composition is sub-divided in unit dose containing appropriate quantities of the active ingredient; the unit dosage forms can be packaged compositions, for example, packeted powders, vials, ampoules, prefilled syringes or sachets containing liquids. The unit dosage form can be, for example, a capsule or tablet itself, or it can be the appropriate number of any such compositions in package form. The dosage to be used in the treatment must be subjectively determined by the attending physician.

In addition, the compounds of this invention may be employed as a solution, cream, or lotion by formulation with pharmaceutically acceptable vehicles containing 0.1-5 percent, preferably 2%, of active compound which may be administered to a fungally affected area.

Accordingly this invention also provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier· The following examples illustrate the preparation of representative compounds of this invention.

AHP-9580/9675-1-N1 - 11 Example 1 Rapamycin-42-ester with N-IY 1.1 -dimethylethoxy)carbonyll-glycylglycine a 7 Under anhydrous conditions, a solution of rapamycin (3 g, 3.28 mmole) and N-[(l,l-dimethylethoxy)carbonyl]-glycylglycine (3.04 g, 13.1 mmole) in 40 mL of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide (1.35 g, 6.56 mmole) followed by 4-dimethylaminopyridine (0.8 g, 6.56 mmole). After stirring at ambient temperature for 48 hours, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were absorbed directly onto silica gel Merck 60 by adding the gel and evaporation to dryness. Flash chromatography of the preabsorbed material (using a gradient elution with ethylacetate-toluene from 2:1 to 1:0 v/v) afforded 1.05 g (28.3 %) of the title compound isolated as a three quarter toluene solvate, along with the 31,42-diester of Example 2. HPLC analysis showed that the monoester is a 8.3:1 mixture of two conformers. !h NMR (CDC13, 400 MHz): δ 1.46 (m, 9H, COOBu*), 1.654 (s, 3H, CH3C=C), 1.751 (s, 3H, CH3C=C), 3.14 (s, 3H, CH3O), 3.33 (s, 3H, CH3O), 3.36 (s, 3H, CH3O), 4.18 (d, IH, CHOH), 4.75 (m, IH, 42-CHO ), 4.79 (s, IH, OH); High Res. MS (neg. ion FAB) Calcd for C6qH93N3O[7: 1127.6504, measured mass 1127.6474.

Anal. Calcd for C6oH93N30I7 · 0.75 PhCH3: C, 65.45; H, 8.33; N, 3.51 Found: C, 65,23; H, 8.32; N, 3.86 The following representative compounds can be prepared from rapamycin and the appropriate terminally-N-substituted amino acid by employing the method used to prepare the title compound in Example 1. b )Rapamycin-42-ester with N-[(fluorenylmethoxy)carbonyl]-alanylserine c) Rapamycin-42-ester with N-[(fluorenylmethoxy)carbonyl]-glycylglycine d) Rapamycin-42-ester with N-[(ethoxy)carbonyl]-arginylmethionine e) Rapamycin-42-ester with N-[(4'-chlorophenoxy)carbonyl]-histidylarginine f) Rapamycin-42-ester with N-[(phenoxy)carbonyl]-tryptophanylleucine g) Rapamycin-42-ester with N-[(phenylmethoxy)carbonyl)]-N-methylglycyl-Nethylalanine AHP-9580/9675-1-N1 - 12h)Rapamycin-42-ester with N-[(phenylmethoxy)carbonyl]-N-methyl-3alanylphenylalanine Rapamycin-42-ester with N-[( 1,1 -dimethylethoxy)carbonyl]-cysteinylglycine Example 2 Rapamvcin-31.42-diester with N-id. l-dimethylethoxv Icarbonyll-glycylglvcine a) The title compound (1.85 g, 42%) was separated from the 42-monoester as described in Example 1 and isolated as a three quarter toluene solvate. HPLC analysis showed that the diester is a 8.1:1 mixture of conformers. !h NMR (CDCI3, 400 MHz): δ 1.452 (m, 18H, COOBu*), 1.6612 (s, 3H, CH3C=C), 1.7815 (s, 3H, CH3C=C), 3.14 (s, 3H, OCH3), 3.34 (s, 3H, OCH3), 3.35 (s, 3H, OCH3), 4.52 (s, 1H, OH), 4.79 (m, 1H, 42-CHO ); High Res. MS (neg. ion FAB): Calcd for C69H107N5O21 1341.7458, measured mass: 1341.7463.

Anal. Calcd for C69H107N5O2I · 0.75 PhCH3: C, 63.17; H, 8.06; N, 4.96 Found: C, 62.83; H, 8.09; N, 5.00 The following representative compounds can be prepared from rapamycin and the appropriate terminally-N-substituted amino acid by employing the method used to prepare the title compound in Example 2. b) Rapamycin-31,42-diester with N-[(fluorenylmethoxy)carbonyl]-alanylserine c) Rapamycin-31,42-diester with N-[(fluorenylmethoxy)carbonyl]-glycylglycine d) Rapamycin-31,42-diester with N-[(ethoxy)carbonyl]-arginylmethionine e) Rapamycin-31,42-diester with N-[(4'-chlorophenoxy)carbonyl]-histidylarginine f) Rapamycin-31,42-diester with N-[(phenoxy)carbonyl]-tryptophanylleucine g) Rapamycin-31,42-diester with N-[(phenylmethoxy)carbonyl)]-N-methylglycyl-Nethyl-alanine h) Rapamycin-31,42-diester with N-[(phenylmethoxy)carbonyl]-N-methyl-βalanylphenyl- alanine i) Rapamycin-31,42-diester with N-[(l,l-dimethylethoxy)carbonyl]-cysteinylglycine AHP-9580/9675-1-N1 - 13Example 3 Rapamycin-31,42-diester with N-i(l,l-dimethylethoxy)carbonyll-N-methylglycine a) Under anhydrous conditions, an ice cold solution of rapamycin (2 g, 2.18 mmole) and Na-Boc sarcosine (1.65 g, 8.75 mmole) in 20 ml of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide (1.8 g, 8.7 mmole) followed by 4-dimethylaminopyridine (1 g, 8.7 mmole). After stirring overnight at ambient temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were evaporated to dryness to give an amorphous amber solid (3 g). The crude product was purified by flash chromatography ( on silica Merck 60, elution with hexane-ethylacetate 1:1, v/v) to provide the title compound (0.75 g, 27.4%) along with the 42-monoester of Example 4. HPLC analysis showed that the diester is a 19.8:1 mixture of two conformers. The multiplicity of the NMR peaks suggests the presence of amide rotamers. !h NMR (CDCI3, 400 MHz): δ 1.411, 1.438, 1.448 and 1.474 (m, 18 H, COOBu1), 2.91 (m, 6H, NCH3), 3.14 (s, 3H, CH3O), 3.34 (s, 3H, CH3O), 3.37 (s, 3H, CH3O), 4.73 (broad, 1H, 42-CHO), 4.82 (2s, 1H, OH); High Res. MS (neg. ion FAB): Calcd. for C67H105N3O19 1255.7342, measured mass 1255.7289.

Anal. Calcd for C67H105N3O19: C, 64.04; H, 8.42; N, 3.34 Found: C, 64.14; H, 8.74; N, 3.63 The following representative compounds can be prepared from rapamycin and the appropriate terminally-N-substituted amino acid by employing the method used to prepare the title compound in Example 3. b) Rapamycin-31,42-diester with N-[(ethoxy)carbonyl]-tyrosine c) Rapamycin-31,42-diester with N-[(fluorenylmethoxy)carbonyl]-phenylalanine d) Rapamycin-31,42-diester with N-[(3',4',5'-trihydroxyphenoxy)carbonyl]-isoleucine e) Rapamycin-31,42-diester with N-[(l,l-dimethylethoxy)carbonyl)-glutamine f) Rapamycin-31,42-diester with N-[(phenoxy)carbonyl]-N-methylalanine g) Rapamycin-31,42-diester with N-[(propyloxy)carbonyl]-4-aminobutryic acid h) Rapamycin-31,42-diester with N-[(phenylmethoxy)carbonyl]-7-aminoheptanoic acid i) Rapamycin-31,42-diester with N-[(fIuorenyImethoxy)carbonyl]-serine AHP-9580/9675-1-N1 - 14Example 4 Rapamycin-42-ester with N-f(L 1-dimethylethoxv icarbonyll-N-methylglycine a) Under anhydrous conditions, an ice cold solution of rapamycin (0.95 g, 1.02 mmole) and Na-Boc sarcosine (0.21 g, 1.1 mmole) in 20 mL of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide 0.21 g, I mmole) followed by 4-dimethylaminopyridine (0.12 g, 1 mmole). After stirring for 4 hours at ambient temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were concentrated in vacuo to give an amorphous amber solid. Flash chromatography of the crude product (on silica Merck 60, elution with hexaneethylacetate 1:1 v/v to remove the diester of Example 3, followed by chloroformethylacetate-methanol 75:25:1 v/v) provided partially purified title compound (0.38 g, 35%). Pure product was obtained by preparative HPLC (Waters Prep 500, silica gel, chloroform-ethylacetate-methanol 75:25:1 v/v, flow rate 250 mL/min). HPLC analysis showed that the ester is a 6.6:1 mixture of two conformers. The multiplicity of NMR peaks suggests the presence of amide rotamers. lH NMR (CDCI3, 400 MHz): δ 1.42-1.46 (ds, 9H, COOBut), 2.91 (ds, 3H, NCH3), 1.644 (s, 3H, CH3C=C), 1.738 (s, 3H, CH3C=C), 3.12 (s, 3H, CH3O), 3.32 (s, 3H, CH3O), 3.35 (s. 3H, CH3O), 4.18 (d, IH, CHOH), 4.71 (broad, IH, 42-CHO), 4.78 (broad s, IH, OH); High Res. MS (neg. ion FAB): Calcd for C59H92N2O16 1084.6446, measured mass 1084.6503.

Anal. Calcd for C59H92N2O16: C, 65.29; H, 8.54; N, 2.58 Found: C, 65.25; H, 8.52; N, 2.42 The following representative compounds can be prepared from rapamycin and the appropriate terminally-N-substituted amino acid by employing the method used to prepare the title compound in Example 4. b) Rapamycin-42-ester with N-[(ethoxy)carbonyl]-tyrosine c) Rapamycin-42-ester with N-[(fluorenylmethoxy)carbonyl]-phenylalanine d) Rapamycin-42-ester with N-[(3',4',5'-trihydroxyphenoxy)carbonyl]-isoleucine e) Rapamycin-42-ester with N-[(l,l-dimethylethoxy)carbonyl)-glutamine f) Rapamycin-42-ester with N-[(phenoxy)carbonyl]-N-methylalanine g) Rapamycin-42-ester with N-[(propyloxy)carbonyl]-4-aminobutryic acid h) Rapamycin-42-ester with N-l(phenylmethoxy)carbonyl]-7-aminoheptanoic acid AHP-9580/9675-1-NI - 15i) Rapamycin-31,42-diester with N-[(fluorenylmethoxy)carbonyl]serine Example 5 Rapamycin-31.42-diester with 5-(T.l-dimethylethoxy)-2-flYl,l-dimethylethoxy)carbonvllaminol-5-oxopentanoic acid a) Under anhydrous conditions, an ice cold solution of rapamycin (4 g, 4.37 mmole) and L-glutamic acid Na_Boc-y-tert-butylester (4.9 g, 16.1 mmole) in 40 mL of dry dichloromethane was treated with dicyclohexylcarbodiimide (1.8 g, 8.7 mmole) followed by 4-dimethylaminopyridine (1 g, 8.7 mmole). After stirring overnight at room temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were concentrated in vacuo to provide 11 g of an amorphous amber solid. The crude product was purified by flash chromatography (on silica Merck 60, gradient elution with hexane-ethylacetate from 2:1 to 1:1, v/v) to yield 4.52 g (69.6%) of the title compound along with the 42-monoester of Example 6. HPLC analysis showed that the diester consists of a 6.6:1 mixture of two conformers. !h NMR (CDCI3, 400 MHz): δ 1.42 (m, 36 H, COOBu1), 1.646 (s, 3H, CH3C=C), 1.701 (s, 3H, CH3C=C), 3.13 (s, 3H, CH3O), 3.34 (s, 3H, CH3O), 3.36 (s, 3H, CH3O), 4.735 (m, 2H, OH+42-CH-O); High Res. MS (neg. ion FAB): calc, for C79Hj25N3O23 1483.8715, measured mass 1483.8714.

Anal. Calcd for C79H125N3O23: C, 63.90; H, 8.49; N, 2.83 Found: C, 63.63; H, 8.41; N, 2.44 The following representative compounds can be prepared from rapamycin and the appropriately terminally-N-substituted amino diacid monoester by employing the method used to prepare the title compound in Example 5. b) Rapamycin-31,42-diester with 6-(phenylmethoxy)-2-[[fluorenylmethoxy)carbonyl]amino]-6-oxohexanoic acid c) Rapamycin-31,42-diester with 6-(4'-methylphenoxy)-3-[[(phenylmethoxy)carbonyl]amino-6-oxohexanoic acid d) Rapamycin-31,42-diester with 6-(ethoxy)-4-[[(phenoxy)carbonyl]amino]-6-oxo35 hexanoic acid AriP-9580/9675-l-Nl - 16e) Rapamycin-31,42-diester with 6-(methoxy)-5-L[(ethoxy)carbonyl]amino]-6-oxohexanoic acid f) Rapamycin-31,42-diester with 4-(phenoxy)-2-[N-[( 1,1 -dimethylethoxy)carbonyl]-Nmethy lamino]-4-oxobutanoic acid g) Rapamycin-31,42-diester with 4-(phenylmethoxy)-3-[N-[(methoxy)carbonyl]-Nmethy lamino]-4-oxobutanoic acid Example 6 Rapamycin-42-ester with 5-(Ll-dimethylethoxy)-2-[f(l.l-dimethylethoxy)carbonynaminol-5-oxopentanoic acid a) The title compound (1.14 g, 20.6%) was separated from the 31,42-diester as 15 described in Example 5 and isolated as the quarter hydrate/mono-ethyl acetate solvate.

HPLC analysis showed that the monoester is a 11.5:1 mixture of two conformers. iH NMR (CDCI3, 400 MHz): δ 1.425 (m, 18H, COOBu1), 1.643 (s, 3H, CH3C=C), 1.737 (s, 3H, CH3C=C), 3.13 (s, 3H, CH3O), 3.32 (s, 3H, CH3O), 3.36 (s, 3H, CH3O), 4.17 (d, 1H, CtfOH), 4.71 (Μ, 1H, 42-CHO), 4.785 (s, 1H, OH); High Resolution MS ( neg. ion FAB): Calc, for C65H1Q2N2O18 1198.7127, measured mass 1198.7077.

Anal. Calcd for ¢¢5^02^018 · CH3COOEt · 0.25 H2O: C, 64.13, H, 8.60; N, 2.17 Found: C, 64.18; H, 8.52: N, 2.01 The following representative compounds can be prepared from rapamycin and the appropriately terminally-N-substituted amino diacid monoester by employing the method used to prepare the title compound in Example 6. b) Rapamycin-42-ester with 6-(phenylmethoxy)-2-[[fluorenylmethoxy)carbonyl]-amino]6-oxohexanoic acid c) Rapamycin-42-ester with 6-(4’-methylphenoxy)-3-[[(phenylmethoxy)carbonyl]-amino35 6-oxohexanoic acid AHP-9580/9675-1-N1 - 17 d )Rapamycin-42-iester with 6-(ethoxy)-4-[[(phenoxy)carbonyl]amino]-6-oxo- hexanoic acid e) Rapamycin-42-ester with 6-(methoxy)-5-[[(ethoxy)carbonyl]amino]-6-oxo- hexanoic acid f) Rapamycin-42-ester with 4-(phenoxy)-2-[N-[(l,l-dimethyIethoxy)carbonyl]-Nmethylamino]-4-oxobutanoic acid g) Rapamycin-42-ester with 4-(phenylmethoxy)-3-[N-[(methoxy)carbonyl]-Nmethylamino]-4-oxobutanoic acid Example 7 Rapamycin-31.42-diester with 2-ii(l.l-dimethylethoxy)carbonvnaminol-4-oxo-4(phenylmethoxv) butanoic acid Under anhydrous conditions, 295mg (1.21mmol) of 2,4,6 trichlorobenzoyl chloride was added to a solution of 39 lmg( 1.21 mmol) of Na-Boc-L-aspartic acid-βbenzyl ester and 170pL (1.21 mmol) of Et3N in 1 mL of THF at room temperature. After stirring for 30 minutes, 500 mg (0.55mmol) of rapamycin and 295 mg ( 2.42 mmol) of dimethylaminopyridine was added and the reaction was left to stir overnight.

The reaction mixture was then filtered and the filtrate concentrated in vacuo. Pure product (200 mg, 25%) was obtained by preparative HPLC (5 cm column, 40 % ethyl acetate-hexane). The product was isolated as the heptahydrate.

JH NMR (CDC13, 400 MHz) δ 7.347 (s, 10 H, Ar), 6.223, 5.126 (s, 4 H, CH2Ph), 4.698 (m, 1 H, CH-CO2), 4.587 (m, 2 H, NH), 3.353 (s, 3 H, CH3O), 3.337 (s, 3 H, CH3O), 3.301 (s, 3 H, CH3O), 2.775 (m, 4 H, CH2CO2); IR (KBr) 3420 (OH), 2935 (CH), 2920 (CH), 1730 (C=O), 1650, 1500, 1455, 1370, 1170 cm-i; MS (neg. ion FAB) 1523 (M‘), 1433, 297, 248, 205, 148, 44, 25 (100).

Anal. Calcd for C83Hi 17N3O23-7H2O C, 60.40; H, 7.09; N, 2.54 Found: C, 60.54; H, 7.28; N, 2.56 AHP-9580/9675-1-N1 - 18Example 8 Rapamycin-31.42-diester with 3-11( Ll-dimethylethoxy)carbonyllaminol-4-oxo-4(phenylmethoxy) butanoic acid 5 Under anhydrous conditions, 532 mg (2.18 mmol) of 2,4,6 trichlorobenzoyl chloride in 1 mL THF was added to a solution of 704 mg (2.18 mmol) of Na-Boc-Laspartic acid-a-benzyl ester and 303 pL (2.18 mmol) of Et3N in 5 mL of THF at room temperature. After stirring for 20 minutes, the reaction mixture was filtered over sintered glass, and the precipitate was washed with THF. The filtrate was concentrated in vacuo to give a thick oil. The oil was dissolved in 5 mL of benzene and LOO g (1.09 mmol) of rapamycin and 532 mg (4.36 mmol) of dimethylaminopyridine in 1 mL of benzene was added dropwise. The reaction was stirred for 2 hr, poured into ethyl acetate, and washed consecutively with 0.5 N HCl and brine. The solution was dried over sodium sulfate, decanted, concentrated in vacuo to give a white foamy solid, which was purified via flash chromatography on a 60 mm x 100 mm silica column (2040 % ethyl acetate/hexane as eluant) to give 532 mg (33 %) of the title compound which was isolated as the hydrate.

!H NMR (CDC13, 400 MHz) δ 7.362 (s, 10 H, Ar), 5.193 (s, 4 H, CH2Ph), 4.596 (tn, 1 H, Ctf-CO2), 4.586 (m, 2 H, NH), 3.336 (s, 3 H, CH3O), 3.306 (s, 3 H, CH3O), 3.145 (s, 3 H, CtfjO); IR (KBr) 3410 (OH), 2950 (CH), 2920 (CH), 1735 (C=O), 1710 (C=O), 1640, 1490, 1445, 1350, 1150 cm -1; MS (neg. ion FAB) 1524 (M-), 1434, 297, 248, 232, 214, 205, 167, 148, 42 (100), 26.

Anal. Calcd for C83Hi 17N3O23 · H2O: C, 65.38; H, 7.73; N, 2.76 Found: C, 64.85; H, 7.67; N, 2.56 Example 9 Rapamycin-42-ester with 3Ti(l.l-dimethylethoxy)carbonynaminol-4-oxo-4(phenylmethoxy) butanoic acid The title compound (374 mg, 23%) was prepared by the method described in the previous Example and separated from the compound described in the previous Example by flash chromatography (20-40% ethyl acetate/hexane as the eluant) and isolated as the sesquihydrate.

AHP-9580/9675-1-N1 - 19iH NMR (CDCI3, 400 MHz) δ 7.356 (s, 5 H, Ar), 5.185 (s, 2 H, CH2Ph), 4.635 (m, 1 H, C//-CO2), 4.582 (m, 1 H, NH), 3.330 (s, 6 H, CH3O), 3.135 (s, 3 H, CtfjO); IR (KBr) 3410 (OH), 2950 (CH), 2920 (CH), 1735 (C=O), 1710 (C=O), 1640, 1490, 1445, 1350, 1150 cm -1; MS (neg. ion FAB) 1218 (M‘), 1127, 590, 168, 42, 25, 17 (100).

Anal. Calcd for C67H98N2Oi8 · 1.5 H2O: C, 63.64; H, 8.21; N, 2.22 Found: C, 63.64; H, 7.51; N, 2.13 Example 10 Rapamycin-42-ester with 5-(1,1 -dimethyloxy)-4-ff(1.1 -dimethylethoxylcarbonvllaminol-5-oxopentanoic acid Under anhydrous conditions, an ice cold solution of rapamycin (4 g, 4.37 15 mmole) and L-glutamic acid Na-Boc-a-tert-butylester (4.9 g, 16.1 mmole) in 40 mL of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide (1.8 g, 8.7 mmole) followed by 4-dimethylamino pyridine (1 g, 8.7 mmole). After stirring overnight at ambient temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were concentrated in vacuo to give 9 g of an amorphous amber solid. The crude product was purified by flash chromatography (on silica Merck 60, gradient elution with hexane-ethylacetate from 2:1 to 3:2, v/v) to provide 1.35 g (25.7%) of the title compound along with the 31,42-diester of Example 11. HPLC analysis showed that the monoester is a 7.5 :1 mixture of two conformers. !h NMR (CDCI3, 400 MHz): δ 1.43 (s, 9H, COOBu1) and 1.46 (s, 9H, COOBu1), 1.65 (s, 3H, CH3C=C), 1.75 (s, 3H, CH3C=C), 3.14 (s, 3H, CH3O), 3.34 (s, 3H, CH3O), 3.38 (s, 3H, CH3O), 4.18 (d, IH, C//-OH), 4.65 (m, IH, 4230 CHO), 4.80 (s, IH, OH); High Res. MS (neg. ion FAB): Calc, for θ65Ηιο2Ν2θ18: 1198.7126, measured mass 1198.7135.

Anal. Calcd for C65H102N2O18: C, 65.09; H, 8.57; N, 2.34 Found C, 65.04; H, 8.33; N, 2.64 AHP-9580/9675-1-N1 -20Example 11 Rapamycin-31.42-diester with 5-(i.l-dimethylethoxy)-4-ri(l.l-dimethylethoxy')carbonvll- aminol-5-oxopentanoic acid 5 The title compound was prepared (0.83 g, 12.8%) along with the 42monoester as described in Example 10. HPLC analysis showed that the diester is a 7.7.T mixture of two conformers.

JH NMR (CDC13, 400 MHz): δ 1.43 (s, 18H, COOBu1), 1.46 (s, 18H, COOBu1), 1.659 (s, 3H, CH3C=C), 1.759 (s, 3H, CH3C=C), 3.14 (s, 3H, CH3O), 3.34 (s, 3H, CH3O), 3.38 (s, 3H, CH3O), 4.66 (m, 1H, 42-CHO), 4.72 (s, 1H, OH); High Res. MS (neg. ion FAB): Calcd for θ79Ηΐ25Ν3θ23: 1483.8704, measured mass 1483.8636.

Anal. Calcd for C79Hi25N3O23: C, 63.90; H, 8.49; N, 2.83 Found: C, 63.68; H, 8.60; N, 3.20 Example 12 Rapamycin-42-ester with Nfi. NE-bisKLl-dimethylethoxykarbonyll-L-lysine Under anhydrous conditions, a solution of rapamycin (3 g, 3.28 mmole) and Na, Ne-bis-Boc-L-lysine (4.5 g, 13 mmole) in 40 mL of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide (1.35 g, 6.56 mmole) followed by 4dimethylaminopyridine (0.8 g, 6.56 m mole). After stirring overnight at ambient temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were concentrated in vacuo to give an amorphous amber solid. Flash chromatography of the crude product (on silica Merck 60, elution with hexaneethylacetate 1:1 v/v) gave partially purified title compound. Pure product (0.8 g, 19.6%) was obtained by preparative HPLC (Waters Prep 500, silica gel, hexaneethylacetate 3:2 v/v, flow rate 250 mL/min). HPLC analysis showed that the monoester is a 9:1 mixture of two conformers.

AHP-9580/9675-1-N1 -21 iH NMR (CDCI3, 400 MHz): δ 1.438 (m, 9H, COOBu1), 1.455 (s, 9H, COOBu1), 1.652 (s, 3H, CH3C=C), 1.752 (s, 3H, CH3C=C), 3.14 3.33(s, 3H, CH3O), 3.37 (s, 3H, CH3O), 4.18 (d, 1H, CHOH), 4.72 (m, 1H, 42CHO), 4.79 (s, 1H, OH); High Res. MS (neg. ion FAB): Calcd for C67H1Q7N3O18: 1241.7549, measured mass 1241.7604.

Anal. Calcd for C^H^^Ois : C, 64.76; H, 8.68; N, 3.38 Found: C, 64.58; H, 9.01; N, 3.10 Example 13 Rapamvcin-31.42-diester with NS, N£-bisl(l,l-dimethylethoxy)carbonyl1-L-lysine Under a nitrogen atmosphere, a solution of Να,Νε bis-Boc-L-lysine (1.038 g, 15 3 mmole) and triethylamine (0.42 mL, 3 mmmole) in 10 mL of anhydrous THF was treated in one portion with 2,4,6-trichlorobenzoyl chloride (0.73 g, 3 mmole). After stirring for 20 minutes at ambient temperature, the precipitated solid was collected and the filtrate was concentrated in vacuo . The resulting mixed anhydride was dissolved in 5 mL of benzene and added to a stirred solution of rapamycin (1 g, 1.09 mmole) containing 4-dimethylamino pyridine (0.59 g, 4.8 mmole) in 10 mL of benzene. After stirring at ambient temperature overnight, the precipitated solid was collected and the filtrate was evaporated to dryness (yellow foam). The crude product was purified by flash chromatography (on silica Merck 60, elution with hexane-ethylacetate 1:1) to provide title compound (1.15 g, 67%). HPLC analysis shows that the diester is a 9:1 mixture of two conformers. iH NMR (CDCI3, 400 MHz): δ 1.426 (m, 9H, COOBuO, 1.438 (s, 9H, COOBu1), 1.443 (s, 9H, COOBu1), 1.446 (s, 9H, COOBu1), 3.141 (s, 3H, CH3O), 3.36 (s, 3H, CH3O), 3.378 (s, 3H, CH3O), 4.68-4.76 (m, 2H, OH and 42-CHO); High res. MS (neg. ion FAB): Calcd. for C83H135N5O23 1569.9526, measured mass 1569.9537.

Anal. Calcd. for C83H135N5O23: C, 63.46; H, 8.66; N, 4.46 Found: C, 63.06; H, 8.84; N, 4.09 AHP-9580/9675-1-N1 -22Example 14.

Rapamycin-14.31.42-tris(monobenzylsuccinate) To a solution of 5.0 g (5.47 mmol) of rapamycin, 3.41 g (16.41 mmol) of monobenzylsuccinate, and 3.15 g (16.41 mmol) of l-(3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride in 20 mL of dry dichloromethane was added 200 mg of 4-dimethylaminopyridine. The solution was stirred at room temperature for 3 days. The reaction mixture was poured into 2 N HCl and extracted three times with ethyl acetate. The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a light yellow foam. Flash chromatography on a 60 mm x 150 mm silica gel column eluting with 20 % ethyl acetate/hexane to 75 % ethyl acetate/hexane gave three fractions. Fraction #1, upon concentration, gave 330 mg (4.1 %) of pure rapamycin-14,31,42-tris15 (monobenzylsuccinate).

NMR (CDC13, 400 MHz) δ 7.353 (bs, 15 H, arom), 5.168 (d, J = 2.0 Hz, 1 H, C//-O2C), 5.148 (m, 6 H, CH2Ph), 4.672 (m, 1 H, CO2C//-CHOMe), 3.355 (s, 3 H, CH3O-), 3.337 (s, 3 H, CH3O-), 3.327 (s, 3 H, CH3O-), 2.697 ( m, 12 H, O2CC7/2C//2CO2CH2Ph), 1.745 (s, 3 H, CH3C=C), 1.655 (s, 3 H, CH3C=C)·, IR (KBr) 3450 (OH), 2950 (CH), 1745 (C=O), 1650, 1460, 1385, 1360, 1160, 1105, 995 cm’1.

Analysis Calcd for C84Hio9N02i-3 H20 C 66.27; H 7.56; N 0.92 Found C 65.96; H 7.24; N 1.00 The following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 14. b) Rapamycin-14,31,42-tris (monomethylsuccinate) c) Rapamycin-14,31,42-tris (monophenyl-3',3'-dimethylglutarate) d) Rapamycin-14,31,42-tris (mono t-butyl-3'-methylglutarate) e ) Rapamycin-14,31,42-tris (monobenzylthiodiglycolate) f) Rapamycin-14,31,42-tris (monohexyldiglycolate) 9) Rapamycin-14,31,42-tris (monopropylphthalate) Rapamycin-14,31,42-tris (monoethyl-2',6'-pyridinedicarboxylate) AHP-9580/9675-1-N1 -23Example 15.

Rapamycin-31.42-bis(monobenzylsuccinate) a 1 Fraction # 2, obtained from the procedure employed in Example 14, gave 1.25 g (17.7 %) of pure rapamycin-3l,42-bis(monobenzylsuccinate) upon concentration. Ή NMR (CDC13, 400 MHz) δ 7.351 (bs, 10 H, arom), 5.168 (d, J = 2.0 Hz, 1 H, C//-O2C), 5.125 (m, 4 H, CH2Ph), 4.680 (m, 1 H, CChC/Z-CHOMe), 3.356 (s, 3 H, CH3O-), 3.329 (s, 3 H, CH3O-), 3.146 (s, 3 H, CH3O-), 2.639 ( m, 8 H, O2CCH2CH2CO2CH2Ph), 1.748 (s, 3 H, C7/jC=C), 1.654 (s, 3 H, C//jC=C); IR (KBr) 3450 (OH), 2940 (CH), 1740 (C=O), 1650, 1455, 1380, 1355, 1160, 1105, 995 cm'1; MS (neg. ion FAB) 1294 (M-), 1202, 1103, 1012, 590, 511, 475, 297, 207, 167, 148, 99 (100); High Res. MS (neg. ion FAB) Calcd for C73H99NO19 1293.68108, found 1293.6811.

Analysis Calcd for C73H99NO19 H20 C 66.82; H 7.70; N 1.07 Found C 67.17; H 7.67; N 1.23 The following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 15. b) Rapamycin-31,42-bis (monomethylsuccinate) c) Rapamycin-31,42-bis (monophenyl-3',3'-dimethylglutarate) d) Rapamycin-31,42-bis (mono t-butyl-3'-methylglutarate) e ) Rapamycin-31,42-bis (monobenzylthiodiglycolate) f) Rapamycin-31,42-bis (monohexyldiglycolate) g) Rapamycin-31,42-bis (monopropylphthalate) h) Rapamycin-31,42-bis (monoethyl-2',6'-pyridinedicarboxylate) Example 16.

Rapamvcin-42-(monobenzylsuccinate) a) Fraction # 3, obtained from the procedure employed in Example 14, gave 930 mg (15.4 %) of pure rapamycin-42-monobenzylsuccinate upon concentration.

AHP-9580/9675-1-N1 -24Hl NMR (CDCI3, 400 MHz) δ 7.355 (bs, 5 H, arom), 5.141 (m, 2 H, CW2Ph), 4.680 (m, 1 H, CO2C//-CHOMe), 3.364 (s, 3 H, CH3O-), 3.333 (s, 3 H, CH3O-), 3.141 (s, 3 H, CH3O-), 2.698 ( m, 4 H, O2CCH2CH2CO2CH2Ph), 1.751 (s, 3 H, CH3C=C), 1.655 (s, 3 H, C//jC=C); IR (KBr) 3450 (OH), 2940 (CH), 1740 (C=O), 1645, 1455, 1380, 1165, 1105, 990 cm’1; MS (neg. ion FAB) 1103 (M-), 1045, 1012, 624, 590, 167, 99 (100); High Res. MS (neg. ion FAB) Calcd for C62H89NO16 1103.6181, found 1103.6048.

Analysis Calcd for C62H89NO16 H20 C 66.36; H 8.02; N 1.24 Found C 66.02; H 7.69; N 1.26 The following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 16. b) Rapamycin-42-(monomethylsuccinate) c) Rapamycin-42-monophenyl-3',3'-dimethylglutarate) d) Rapamycin-42-(mono t-butyl-3'-methylglutarate) e) Rapamycin-42-(monobenzylthiodiglycolate) f 1 Rapamycin-42-(monohexyldiglycolate) 9) Rapamycin-42-(monopropylphthalate) h 1 Rapamycin-42-(monoethyl-2',6'-pyridinedicarboxylate) Example 17.

Rapamycin-31.42-bishemiglutarate a) To a solution of 2.0 g (2.2 mmol) of rapamycin in 10 mL of dry dichloromethane was added 1.24 g (10.9 mmol) of glutaric anhydride followed by 881 uL (861 mg, 10.9 mmol) of pyridine. To this was added 200 mg of 4-dimethylaminopyridine and the reaction mixture was allowed to reflux for 8 h. The solution was cooled to room temperature, poured into 2 N HCI, and extracted three times with dichloromethane. The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow foam. The crude product was purified via reverse phase HPLC on a Ci8 column eluting starting with 60 % acetonitrile/water. Collected, after, concentration, 586 mg (24 %) of rapamycin-31,42-bishemiglutarate.

AHP-9580/9675-1-N1 -25NMR (CDC13, 400 MHz) δ 5.398 (m, 1 H, -CC^CHCHOMe), 4.683 (m, 1 H, -CO2CHCHOMe), 3.364 (s, 3 H, C//3O-), 3.362 (s, 3 H, CH3O-), 3.106 (s, 3 H, C//3O-), 2.407 (m, 8 H, -O2CC//2CH2CH2CO2H), 1.960 (m, 4 H, -O2CCH2C//2CH2CO2H), 1.770 (s, 3 H, CH3C=O, 1.653 (s, 3 H, CH3C=C); 13C NMR (CDCI3, MHz) 211.45 (C=O), 206.84 (C=O), 200.44 (C=O), 177.83 (C=O), 177.04 (C=O), 172.43 (C=O), 171.20 (C=O), 165.27 (C=O), 159.08 (C=O); IR (KBr) 3430 (OH), 2940 (CH), 2880 (CH), 1745 (C=O), 1685, 1625, 1580, 1450, 1385, 1330, 1200, 1140, 1100, 990 cm-1; MS (neg. ion FAB) 1140 (M-H), 1122, 1026, 990, 946, 913, 590, 475, 435, 321, 167, 148, 131 (100), 113; High Res.

MS (neg. ion FAB) Calcd for C61H90O19N (M-H) 1140.6107, Found 1140.6106.

Analysis Calcd for C6iH9iO19N H2O C 63.15; H 8.02; N 1.20 Found C 63.35; H 7.88; N 1.40 The following representative compounds can be prepared from rapamycin and the appropriate anhydride by employing the method used to prepare the title compound in Example 17. b) Rapamycin-31,42-bishemi-3'-methylglutarate c ) Rapamycin-3l,42-bishemi-3',3'-dimethylglutarate d) Rapamycin-3 l,42-bishemi-3'-oxoglutarate e) Rapamycin-3 l,42-bishemi-3'-thioglutarate f) Rapamycin-31,42-bishemi-phthalate g) Rapamycin-31,42-bishemi-2',3'-pyridine dicarboxylate Example 18.

Rapamvcin-31.42-hemiglutarate bissodium salt Purified bis-31,42-hemiglutarate of rapamycin (740 mg, 649 umol), prepared as described in Example 17, was dissolved in 5 mL of 95 % ethanol and 107 mg (1.27 mmol) of sodium bicarbonate was added. Water (1 mL) was added to completely dissolve the salt. Once dissolved, the light yellow solution was concentrated in vacuo to give a foamy yellow solid. The foam was dried in a drying pistol for 24 h, refluxing over acetone at reduced pressure to give 520 mg of the bissodium salt.

AHP-9580/9675-1-N1 -26>H NMR (d^-DMSO, 400 MHz) δ 5.235 (m, 1 H, -C//O2C), 4.498 (m, 1 H, MeOCHC7/O2C-), 3.287 (s, 6 H, 2 C//3O-), 3.236 (s, 3 H, CW3O-), 2.245 (m, 8 H, O2CCW2CH2C//2CO2-), 1.712 (s, 3 H, CtfjC=C), 1.593 (s, 3 H, Ctf?C=C); IR (KBr) 3420 (OH), 2920 (CH), 1725 (C=O), 1675, 1620, 1560, 1450, 1400, 1375, 1230, 1195, 1130, 1090, 980 cm'1; MS (neg. ion FAB) 1112 (M-l, free acid), 994, 589, 475, 297, 167, 148, 117, 99 (100); High Res. MS (neg. ion FAB) Calcd for C6iH89Oi9NNa (M-Na) 1162.5926, Found 1162.5899.

Analysis Calcd for C6iH89Oi9NNa2-H2O C 60.85; H 7.56; N 1.16 Found C 60.67; H 7.36; N 1.58 Example 19.

Rapamycin-31.42-bishemiglutarate bistromethamine salt Purified bis-31,42 hemiglutarate of rapamycin (950 mg, 833 umol), prepared as described in Example 17, was dissolved in 5 mL of 95 % ethanol and 197 mg (1.63 mmol) of tris(hydroxymethyl)methylamine was added. Water (1 mL) was added to completely dissolve the amine. Once dissolved, the yellow solution was concentrated in vacuo to give a foamy yellow solid. The very hygroscopic foam was dried in a drying pistol for 24 h, refluxing over acetone at reduced pressure to give 900 mg (78 %) of the bistromethamine salt. Ή NMR (d6-DMSO, 400 MHz) δ 5.253 (m, 1 H, -CHO2C), 4.523 (m, 1 H, MeOCHC7/O2C-), 3.347 (s, 6 H, 2 C//3O-), 3.276 (s, 3 H, C//3O-), 2.289 (m, 8 H, O2CCH2CH2CH2CO2-), 1.681 (s, 3 H, CH3C=C), 1.595 (s, 3 H, CH3C=Cy IR (KBr) 3400 (OH), 2920 (CH), 1730 (C=O), 1620, 1555, 1450, 1400, 1370, 1185, 1060, 980 cm'1; MS (neg. ion FAB) 1140 (M-H, free acid), 1028, 167, 148, 131 (100), 113; High Res. MS (neg. ion FAB) Calcd for C6iH9oOi9N (M-H, free acid) 1140.6107, Found 1140.6069.

Analysis Calcd for C69H103O25N3 · 2 H2O C 58.77; H 7.58; N 2.98 Found C 58.47; H 7.94; N 3.58 AHP-9580/9675-1-N1 -27 Example 20.

Rapamycin-42-hemi-3'-oxoglutarate a) To a solution of 3.0 g (3.3 mmol) of rapamycin in 20 mL of dry dichloromethane was added 1.90 g (16.4 mmol) of diglycolic anhydride followed by 1.32 mL (1.29 g, 16.4 mmol) of pyridine. To this was added 200 mg of 4-dimethylaminopyridine and the reaction mixture was allowed to stir at room temperature for 2 days. The solution was cooled to room temperature, poured into 2 N HCl, and extracted three times with dichloromethane. The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow foam. The crude product was purified via reverse phase HPLC on a Cis column eluting starting with 60 % acetonitrile/water. After concentration, 870 mg ( 26 %) of rapamycin-42-hemi-3'-oxoglutarate and 500 mg (13 %) of rapamycin-31,42-bishemi-3Oxoglutarate were isolated.

!H NMR (CDC13, 400 MHz) δ 4.768 (m, 1 H, CO2C//-CHOMe), 4.250 (m, 4 H, O2CCH2OCH2CO2), 3.356 (s, 3 H, CH3O-), 3.331 (s, 3 H, C//3O-), 3.139 (s, 3 H, CH3O-), 1.759 (s, 3 H, CHjC=C), 1.653 (s, 3 H, C//?C=C); IR (KBr) 3420 (OH), 2920 (CH), 2875 (CH), 1740 (C=O), 1720 (C=O), 1640, 1625, 1445, 1370, 1320, 1200, 1135, 1095, 980 cm'1; MS (neg. ion FAB) 1028 (M - H), 327, 167 (100), 148, 133, 115; High Res. MS (neg. ion FAB) Calcd for C55H82O17N (M - H) 1028.5597, Found 1028.5599.

Analysis Calcd for C55H83O17N 3 H2O C 60.97; H 8.22; N 1.29 Found C 61.33; H 7.74; N 1.69 The following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 20. b) Rapamycin-42-hemi-3'-methylglutarate c) Rapamycin-42-hemi-3’,3'-dimethylglutarate d) Rapamycin-42-hemi-3'-thioglutarate e) Rapamycin-42-hemi-phthalate f) Rapamycin-42-hemi-2',3'-pyridine dicarboxylate AHP-9580/9675-1-N1 -28Example 21.

Rapamvcin-31.42-bishemi-3’-oxoglutarate To a solution of 5.0 g (5.47 mmol) of rapamycin in 20 mL of dry dichloromethane was added 3.17 g (27.3 mmol) of diglycolic anhydride followed by 2.17 mL (2.12 g, 27.3 mmol) of pyridine. To this was added 400 mg of 4-dimethylaminopyridine and the reaction mixture was allowed to stir at reflux for 24 h. The solution was cooled to room temperature, poured into 2 N HCl, and extracted three times with dichloromethane. The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow foam. The crude product was purified via reverse phase HPLC on a Cis column eluting starting with 60 % acetonitrile/water. After concentration, 1.75 g (28 %) of rapamycin-31,42-bishemi-3'-oxoglutarate was isolated.

!H NMR (CDC13, 400 MHz) δ 4.785 (m, 1 H, CO2CHCHOMe), 4.260 (m, 8 H, O2CCH2OCH2CO2), 3.360 (s, 3 H, CH3O-), 3.343 (s, 3 H, CH3Q-), 3.143 (s, 3 H, C//3O-), 1.775 (s, 3 H, CH3C=C), 1.656 (s, 3 H, C7/jC=C); 13C NMR (CDCI3, MHz) 211.12 (C=O), 207.73 (C=O), 193.11 (C=O), 171.90 (C=O), 171.59 (C=O), 170.15 (C=O), 169.35 (C=O), 168.83 (C=O), 166.63 (C=O); IR (KBr) 3420 (OH), 2920 (CH), 2850 (CH), 1740 (C=O), 1645, 1625, 1440, 1370, 1190, 11300, 980 cm'1; MS (neg. ion FAB) 1140 (M-H), 1122, 1026, 990, 946, 913, 590, 475, 435, 321, 167, 148, 131 (100), 113; High Res. MS (neg. ion FAB) Calcd for C59H86O21N (M-H) 1144.5701, Found 1144.5702.

Analysis Calcd for C59H87O2iN C 61.82; H 7.65; N 1.22 Found C 61.59; H 7.36; N 1.84 Example 22.

Rapamycin-31.42-bishemi-3'-oxoglutarate disodium salt Purified bis-31,42 hemi-3'-oxoglutarate of rapamycin (720 mg, 629 umol) , prepared by the procedure employed in Example 21, was dissolved in 10 mL of 95 % ethanol and 106 mg (1.26 mmol) of sodium bicarbonate was added. Water (1 mL) was added to completely dissolve the salt. Once dissolved, the light yellow solution was concentrated in vacuo to give a foamy yellow solid. The foam was dried in a drying AHP-9580/9675-1-N1 -29pistol for 48 h, refluxing over dichloromethane at reduced pressure to give 435 mg (58 %) of the disodium salt.

JH NMR (d^-DMSO, 400 MHz) δ 4.975 (m, 1 H, -CHO2C), 4.593 (m, 1 H, MeOCHC//O2C-), 4.135 (s, 2 H, -O2CCH2OCH2CO2R), 3.617 (s,2 H, O2CCH2OCH2CO2R), 3.299 (s, 6 H, 2 CH3O-), 3.232 (s, 3 H, C//3O-), 1.614 (s, 3 H, CH3C=C), 1.553 (s, 3 H, C//jC=C); IR (KBr) 3420 (OH), 2920 (CH), 1735 (C=O), 1615, 1445, 1395, 1380, 1320, 1220, 1130, 1090, 980 era'1; MS (neg. ion FAB) 1188 (M-l), 1166 (M-Na), 1144, 1051, 1028, 590, 459, 167, 155 (100), 148, 133, 115.

Analysis Calcd for C59H85O2iNNa2 · 2H2O C 57.79; H 7.26; N 1.14 Found C 57.94; H 7.11; N 1.26 Example 23.

Rapamycin-31.42-bishemi-3'-oxoglutarate bistromethamine salt Purified bis-31,42 hemi-3'-oxoglutarate of rapamycin (1.01 g, 882 umol), prepared by the procedure employed in Example 21,was dissolved in 10 mL of 95 % ethanol and 213 mg (1.76 mmol) of tris(hydroxymethyl)- methylamine was added. Water (1 mL) was added to completely dissolve the amine. Once dissolved, the yellow solution was concentrated in vacuo to give a foamy yellow solid. The very hygroscopic foam was dried in a drying pistol for 48 h, refluxing over dichloromethane at reduced pressure to give 805 mg (66 %) of the bistromethamine salt.

*H NMR (d6-DMSO, 400 MHz) δ 4.955 (m, 1 H, -CHO2C), 4.600 (m, 1 H, MeOCHC//O2C-), 4.149 (s, 2 H, -O2CCH2OCH2CO2R), 3.770 (s, 2 H, -O2CC//2OCH2CO2R), 3.407 (s, 6 H, 2 C//3O-), 3.257 (s, 3 H, C//3O-), L806 (s, 3 H, CH3C=C), 1.614 (s, 3 H, CH3C=C)·, IR (KBr) 3400 (OH), 2920 (CH), 1730 (C=O), 1620, 1550, 1450, 1395, 1370, 1200, 1060, 985 cm'1; MS (neg. ion FAB) 1144 (M-H, free acid), 1028, 167, 148, 133 (100), 115.

Analysis Calcd for C67H109O27N3 · H2O C 57.22; H 7.90; N 2.98 Found C 57.26; H 7.90; N 3.15 AHP-9580/9675-1-N1 -30Example 24.

Rapamycin-31.42-bishemisuccinate.

To a solution of 2.0 g (2.2 mmol) of rapamycin in 10 mL of dry dichloromethane was added 1.19 g (10.9 mmol) of succinic anhydride followed by 881 uL (861 mg, 10.9 mmol) of pyridine. To this was added 200 mg of 4-dimethylaminopyridine and the reaction mixture refluxed for 24 h. The solution was cooled to room temperature, poured into 2 N HCI, and extracted three times with dichloromethane.

The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow foam. The crude product was purified via reverse phase HPLC on a Cjs column gradient eluting starting with 20 % acetonitrile/water to 60 % acetonitrile/water. Collected, after, concentration, 770 mg (31 %) of rapamycin-31,42-bishemisuccinate.

The purified bis-31,42 hemisuccinate of rapamycin (770 mg, 686 umol) was dissolved in 10 mL of 95 % ethanol and 166 mg (1.37 mmol) of tris(hydroxymethyl)methylamine was added. Water (1 mL) was added to completely dissolve the amine. Once dissolved, the yellow solution was concentrated in vacuo to give a foamy yellow solid. The very hygroscopic foam was dried in a drying pistol for 24 h, refluxing over acetone at reduced pressure to give 890 mg (95 %) of the bistromethamine salt. The bistromethane salt was evaluated in the standard pharmacological test procedures.

JH NMR (d6-DMSO, 400 MHz) 5.231 (m, 1 H, -CHO2C), 4.554 (m, 1 H, MeOCHCHO2C-), 3.426 (s, 6 H, 2 CH3O-), 3.249 (s, 3 H, C//3O-), 2.431 (m, 8 H, O2CCH2CH2CO2-), L700 (s, 3 H, C//jC=C), 1.554 (s, 3 H, CHjC=C); 13C NMR (d6-DMSO, ) 211.28 (C=O), 205.23 (C=O), 199.59 (C=O), 174.86 (C=O), 173.62 (C=O), 171.72 (C=O), 171.50 (C=O), 166.56 (C=O), 166.53 (C=O); IR (KBr) 3420 (OH), 2940 (CH), 1735 (C=O), 1630, 1580, 1460, 1400, 1380, 1170, 1070, 990 cm-1; MS (neg. ion FAB) 1112 (M-l, free acid), 994, 589, 475, 297, 167, 148, 117, 99 (100).

Analysis Calcd for C67H109O25N3 · 2 H2O C 57.80; H 8.12; N 3.01 Found C 57.91; H 8.21; N 2.37

Claims

1. A compound of the structure wherein R 1 , R 2 ’ and R 3 are each, independently, hydrogen, or R 4 ; O II ? R 4 is — [C(CH 2 ) m CH(CH 2 ) n N] p CO 2 R 7 , -C-fCH^XtCH^CC^R 11 , or I I R 5 R 6 O ll -C-ί- -J—co 2 r 12 . R 5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, -(CH 2 )qCO 2 R 8 , -(CH2)rNR 9 CO2R 10 , carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxy alkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazctylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; AHP-9580/9675-1-N1 -32R 6 and R 9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms; R 7 , R 8 , and R 10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or trisubstituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R 11 and R 12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R 13 Xis -C-, O, orS; R 14 R 13 and R 14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms; Y is CH or N; m is 0 - 4; n is 0 - 4; p is 1 - 2; q is 0 - 4; r is 0 - 4; t is 0 - 4; u is 0 - 4; O II wherein R 5 , R 6 , m, and n are independent in each of the[C(CH 2 ) m CH(CH2) n N] I I R 5 R 6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R 1 , R 2 - and R 3 are not all hydrogen, further provided that R 1 , R 2 > and R 3 are not all AHP-9580/9675-1-N1 -33O II ~[C(CH) CH(CFL) N] CO„R , and still further that 2 m 2 η ι p 2 R 5 R 6 t and u are not both 0 whe»X is 0 or S.

2. A compound of Claim 1 where R is O CH(CH„) N] CO_R 7 , 2 m | 2 η p 2 K R 6 m=0 and n=0, or a pharmaceutically acceptable salt thereof .

3. A compound of Claim 1 or Claim 2 where n=0, and 5 8 R is -(CH 2 ) CO 2 R or a pharmaceutically acceptable salt thereof.

4. A compound of Claim 2 where R^ is 9 10 -(CH^^NR CC^R or a pharmaceutically acceptable salt thereof .

5. A compound of Claim 2 where R^ is hydrogen or a pharmaceutical 1y acceptable salt thereof. 4 .

6. A compound of Claim 1 where R is O Il ii -C-(CH_).X(CH_) CO-R 2 t 2 u 2 or a pharmaceutically acceptable salt thereof.

7. Rapamycin-42-ester with N-[(1,1-dimethylethoxy)carbony1 ]g1ycy1g1ycine or a pharmaceutica11y acceptable salt thereof . ΑΗΡ-9580/9675-1-NI -348. Rapamycin-31,42-diester with N-[(l,ldimethylethoxy1carbony1]-glycy1g1ycine or a pharmaceutically acceptable salt thereof.

8. 9. Rapamycin-31,42-diester with N-[(l,ldimethylethoxylcarbonyl]-N-methylglycine or a pharmaceutically acceptable salt thereof.

9. 10. Rapamycin-42-ester with N-[(l,ldimethylethoxy 1carbony1]-N-methylg1ycine or a pharmaceutically acceptable salt thereof.

10. 11. Rapamycin-3 1,42-diester with 5-(1,1dimethylethoxy)-2-[[(1,l-dimethylethoxy)carbonyl]ami no]-5-oxopentanoic acid or a pharmaceutically acceptable salt thereof.

11. 12. Rapamycin-42-ester with 5-(1,1-dimethylethoxy )2-[[(1,1-dimethylethoxylcarbonyl]amino]-5-oxopentanoic acid or a pharmaceutically acceptable salt thereof.

12. 13. Rapamycin-31,42-diester with 2-[[(1,1dime thylethoxy)carbony1 ]amino]-4-oxo-4 -Cpheny1 methoxy ) butanoic acid or a pharmaceutically acceptable salt thereof .

13. 14. Rapamycin-31,42-diester with 3—E[(1,1— dimethyl ethoxy )carbony1 ]amino]-4-oxo-4-Cphenyl methoxy 1 butanoic acid or a pharmaceutically acceptable salt thereof .

14. 15. Rapamycin-4 2-ester with 3 — Ε E(1,1-dimethy 1 ethoxylcarbonyl]amino]-4-oxo-4(pheny1 methoxy 1 butanoic acid or a pharmaceutically acceptable salt thereof. AHP-9580/9675-1-Nl -3516. Rapamycin-42-ester with 5-(1,1-dimethy1oxy)4 -[[(1,1-dimethylethoxy ) carbonyl ]amino]- 5-oxopentanoic acid or a pharmaceutically acceptable salt thereof.

15. 17. Rapamycin-31,42-diester with 5-(1,1dimethylethoxy )-4-[[(1,1-dimethylethoxylcarbonyl Jamino]-5-oxopentanoic acid or a pharmaceutically acceptable salt thereof.

16. 18. Rapamycin-42-ester with N a , N^bis[(l,ldimethy1 ethoxy )carbonyl ]-L-1ysine or a pharmaceutically acceptable salt thereof.

17. 19. Rapamycin-31,42-diester with Ν α , N c bis[(l,ldimethylethoxyIcarbonyl]-L-lysine or a pharmaceutically acceptable salt thereof.

18. 20. Rapamycin-14,31,42-tris(monobenzy1succinate) or a pharmaceutically acceptable salt thereof.

19. 21. Rapamycin-31,42-bis(monobenzy1succinate1 or a pharmaceutically acceptable salt thereof.

20. 22. Rapamycin-4 2-(monobenzylsuccinate1 or a pharmaceutically acceptable salt thereof.

21. 23. Rapamycin-31,42-bishemiglutarate or a pharmaceutically acceptable salt thereof.

22. 24. Rapamycin-32,42-hemiglutarate bissodium salt.

23. 25· Rapamycin-31,42-bishemiglutarate bistromethamine salt.

24. 26. Rapamycin-42-hemi-3'-oxog1utarate or a pharmaceutically acceptable salt thereof. ΑΗΡ-9580/9675-1-N1 -3627. Rapamycin-31,42-bishemi-3'-oxog1utarate or a pharmaceutically acceptable salt thereof.

25. 28. Rapamycin-31,42-bishemi-3'-oxog]utarate disodium salt.

26. 29. Rapamyc in-31,4 2-bishemi-3'-oxoglutarate bistromethamine salt.

27. 30. Rapamycin-31,42-bishemisuccin ate or a pharmaceutically acceptable salt thereof.

28. 31. Rapamycin-31,42-bishemisuccinate bistromethane salt.

29. 32. A compound as claimed in any one of Claims 1 to 23, 26, 27 and 30 when in the form of a salt of an inorganic cation; a mono-, di- or tri-alkyl amine of 1 to 6 carbon atoms per alkyl group; a mono-, di- or tri-hydroxya 1ky1 amine of 1 to 6 carbon atoms per alkyl group or an acid selected from acetic, lactic, citric, tartaric, succinic, maleic, malonic and gluconic.

30. 33. A process for preparing a compound of formula I as claimed in Claim 1 which comprises Ca) acylating rapamycin with an acylating agent or Cb) sequentially acylating rapamycin with one or more acylating agents, said acylating agentCs) being selected from acids of formula: Z-tCCCH,)CHCCH Q )N] C0 o R 7 2 m. 2 n J p 2 O |l j 1 !i H p p Z-C-(CH_) X(CH_) CClR 11 2 t 2 u 2 and CO 2 R AHP-9580/9675-1-N1 -37wherein Z is OH and the other variables are as defined in Claim 1, or reactive derivatives thereof, if desired protecting any of 42, 31 and 14 positions of rapamycin with an appropriate protecting group and removing said group as required, and further if desired isolating the product as a pharmaceutically acceptable salt.

31. 34. A process as claimed in Claim 33 in which the acylation is carried out using a coupling agent.

32. 35. A process as claimed in Claim 33 in which the reactive derivative in the anhydride, mixed anhydride derived from 2,4,6-trichlorobenzoyl chloride or the acid chloride, bromide or iodide.

33. 36. A process for preparing a compound according to Claim 1 or a pharmaceutically acceptable salt thereof substantially as hereinbefore described and illustrated in any one of Examples 1 a) to il; 2 a) to il; 3 a) to i); 4 al to i); 5 a) to g); 6 a) to gl; 7 to 13; 14 a) to hl; 15 al to h); 16 al to hl; 17 a) to gl; 18, 19, 20 al to fl and 21 to 24.

34. 37. A compound of formula I whenever prepared by a process according to any one of Claims 33 to 36.

35. 38. A pharmaceutical composition comprising a compound of formula I or a-pharmaceutical1y acceptable salt thereof as claimed in any one of Claims 1 to 32 and a pharmaceutically acceptable carrier.