WO1998009970A2

WO1998009970A2 - Alkylated rapamycin derivatives

Info

Publication number: WO1998009970A2
Application number: PCT/US1997/015439
Authority: WO
Inventors: David Cheng Hu; Alexander Aleksey Grinfield; Thomas Joseph Caggiano; Craig Eugene Caufield
Original assignee: American Home Products Corporation
Priority date: 1996-09-09
Filing date: 1997-09-03
Publication date: 1998-03-12
Also published as: JP2001500126A; CN1235608A; EP0927182A2; AU4176897A; WO1998009970A3; CA2266039A1

Abstract

This invention relates to compounds which possess immunosuppressive and/or anti tumor and/or antiinflammatory activity in vivo and/or inhibit thymocyte proliferation in vitro. These compounds are therefore useful in the treatment of transplantation rejection, autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, multiple sclerosis and in the treatment of Candida albicans infections and also in treatment of diseases of inflammation. These compounds are represented by formula (I), wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein: R1 is selected from -(CH¿2?)n-Ar where Ar is not phenyl, -(CH2CH2O)nCH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n-CH2CH(OR?3)CH¿2OR4 where R?3 and R4¿ are H, C¿1?-C10 alkyl, or R?3 and R4¿ together are ethylene, methylene or dimethylmethylene; -CH¿2?(CH2)n-OR?3¿ where R3 is not H, C¿1?-C10 alkyl, or C(O)C1-C10 alkyl; and -CH2(CH2)n-X where X is F, Cl, Br or I; R?2¿ is selected from H, C¿1?-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR?3, -CH¿2(CH2)n-X where X is F, Cl, Br or I; and -(CH2)nCH2CH(OR5)CH2OR6 where R?5 and R6¿ are selected independently from H, C¿1?-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR?7¿ and -CO¿2R?7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar; n = 1-10 independently; m = 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl; 1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.

Description

ALKYLATED RAPAMYCIN DERIVATIVES

This invention relates to compounds of formula I below or pharmaceutically acceptable salts thereof which possess immunosuppressive and/or anti tumor and/or antiinflammatory activity in vivo and/or inhibit thymocyte proliferation in vitro. These compounds are therefore useful in the treatment of transplantation rejection, autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, multiple sclerosis, the treatment of Candida albicans infections, treatment of diseases of inflammation treatment of hyperproliferative vascular disease (restenosis) and in the treatment of certain human tumors.

BACKGROUND OF THE INVENTION

Rapamycin is a tnacrocyclic 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 3,993,749J.

Rapamycin

(Positions numbered according to Chemical Abstracts)

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 5,100,899). 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. Calne et al., Lancet 1 183 (1978)]. U. S. patent 5,321 ,009 discloses a method of prophylactically preventing the onset, preventing the development, and arresting the progression of insulin-dependent diabetes mellitus by administration of rapamycin. U. S. patent 5,288,71 1 discloses a method of preventing or treating hyperproliferative vascular disease by administration of a combination of rapamycin and heparin. U. S. patent 5,286,730 discloses a method of treating immunoinflammatory disease by treatment with rapamycin alone or in combination with cyclosporin A. U. S. patent 5,286,731 provides a method of treating immunoinflammatory bowel disease by administration of rapamycin alone or in combination with cyclosporin A. Various structural features of rapamycin have been modified in efforts to increase the potency or specificity of pharmacological action. For instance, a number of U. S. patents disclose compounds where one or more of the hydroxy groups having normal stereochemistry at positions 14, 31 , and 42 have been converted into acyl esters, sulfonyl esters, and carba ates. U. S. patent 5,023,263 discloses 42-oxo rapamycin. U. S. patent 5,258,389 discloses 31 and/or 42 O-alkyl, O-aryl, O-alkenyl, and O-alkynyl ethers of rapamycin having normal stereochemistry at the 42 position. The PCT published application WO 94/09010 discloses 31 and/or 42 O-alkylate rapamycin analogs wherein the keto groups at positions 15 and 33 may be reduced to a hydroxyl group or a methylene group.

SUMMARY OF THE INVENTION

The compounds of this invention are represented by the following chemical formula and are novel. While a number of rapamycin analogs substituted at positions 31 and 42 have been disclosed, substitution at position 29 has not been heretofore disclosed. The compounds useful in this invention are represented by the formula below:

wherein W and Y are OR' and X and Z together form a bond or W and X are OR² and Y and Z together form a bond, wherein R¹ is selected from -(CH2)_n-Ar where Ar is not phenyl, -(CH₂CH₂O)_nCH3 where n is not 1, -CH₂CH₂CH₂O(CH2CH₂O)_m-CH3,

-(CH₂)n-CH2CH(OR³)CH₂OR⁴ where R³ and R⁴ are H, -Cio alkyl, or R3 and R⁴ together are ethylene, methylene or dimethylmethylene; -CH₂(CH2)_n-OR3 where R³ is not H, Cι-C_l0 alkyl, or C(O)Cι-Cιo alkyl; and -CH₂(CH₂)_n-X where X is F, Cl, Br or I;

R² is selected from H, Cj-Cio alkyl, Ar(CH₂)_n-, C3-C₁0 alkenyl, -(CH₂CH₂O)nCH3, -CH₂CH2CH2θ(CH2CH₂O)_m-CH3, -CH₂(CH₂)_n-OR3, -CH₂(CH₂)_n-X where X is F, Cl, Br or I; and -(CH₂)_n-CH₂CH(OR⁵)CH₂OR⁶ where R⁵ and R⁶ are selected independently from H, Ci-Cjn alkyl, -(CH₂)_n-Ar, -CONH(CH₂)_n-Ar or COC(CH3)2-(CH₂)_n-Ar, -COR7 and -CO₂R⁷, where R? is Cι~C₆ alkyl, C₂-C6 alkenyl, or Ar;

n= 1-10 independently; and m= 1-5 independently;

In the above definitions of variable substituents, the term alkyl includes straight and branched chain hydrocarbons and the aryl groups (Ar) are selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl, 1 , 2, 3-triazolyl and tetrazolyl. This invention also encompasses the pharmaceutically acceptable acid addition salts where they can be formed.

The compounds of this invention exhibit immunosuppressive and/or antifungal and/or antitumor and/or antiinflammatory activity in vivo and/or inhibit thymocyte proliferation in vitro and are therefore useful in the treatment or inhibition of organ or tissue transplantation rejection or host vs. graft disease, proliferative diseases such as restenosis following angioplasty procedures, autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, and multiple sclerosis; fungal infections, and diseases of inflammation such as psoriasis, exzema, seborrhea, inflammatory bowel disease and pulmonary inflammation such as asthma, chronic obstructive pulmonary disease, emphysema, bronchitis and the like. An invention compound was found to inhibit the growth of human breast, colon and ovarian cancer cell lines in submicromolar concentration and it is therefore expected that the invention compounds will be useful in inhibiting these and other tumors in humans.

Detailed Description of the Invention

The compounds of this invention can be prepared by standard literature procedures as outlined below. Other suitable bases may be used in place of the 2,6-di- tert-butyl-4-methylpyridine and may include pyridine, lutidine, collidine, sodium hydride, or sodium carbonate.

+ 31 -substituted product and 31 ,42-disubstituled product Also, unexpectedly, a product was isolated (1,3-allyl rearrangement) via triflation at C-28 followed by attack by external nucleophile.

The compounds of this invention exhibit immunosuppressive and/or antifungal and/or antitumor and/or antiinflammatory activity in vivo and/or inhibit thymocyte proliferation in vitro and are therefore useful in the treatment of transplantation rejection, autoimmune diseases (i.e. lupus, rheumatoid arthritis, diabetes mellitus, multiple sclerosis), Candida albicans infections, and diseases of inflammation. An invention compound inhibits in vitro in sub-micromolar concentrations, the growth of certain human tumor cell lines, including colon (MIP 101), breast (T47D, SKBR-3), and ovarian (A2780S) cells and therefore it is expected that compounds of this inventon will be useful in the treatment of these and other tumors.

The following procedures are included to exemplify the preparation of invention compounds and employ standard laboratory techniques known to those skilled in the art of organic systhesis. Example 1. 29-O-Benzylraμamyein

To a solution of triflic anhydride (0.19 mL, 1.13 mmol) in dichloromethane (5.0 mL) in a round bottom flask (50 mL, flame dried) equipped with a magnetic stirrer at 0 *C was added 2,6-di-t-butyl-4-methyl pyridine (0.253 g, 1.23 mmol) portionwise. The reaction mixture was degassed, and purged with nitrogen. To the solution was added a solution of benzyl alcohol (0.1 1 mL, 1.06 mmol, diluted in 3 mL dichloromethane) dropwise. The solution became a pale white suspension. The reaction was stirred at 0 °C for 30 min. TLC analysis indicated no residual benzyl alcohol remained. To the suspension was added another portion of 2,6-di-t-butyl-4-methyl pyridine (0.335 g, 1.63 mmol) portionwise, followed by a solution of rapamycin (0.855 g, 0.94 mmol, in 3 mL dichloromethane) dropwise. The reaction was stirred at 0 °C for 30 min, and warmed up to room temperature and stirred overnight. The reaction was quenched with saturated aqueous NaHCO3, and the organic and aqueous layers were separated. The aqueous layer was extracted three times with ethyl acetate. The organic layers were combined, washed with brine, and dried over sodium sulfate. The solution was filtered and concentrated in vacuo to afford a pale yellow foam. The product mixture was separated by HPLC (40 % EtOAc/hexane, Dynamax 2" silica column, 20 mL/min), and four fractions were collected. The second fraction which was obtained in 12 % yield was identified as 42-O-benzylrapaιnycin. The third fraction which was obtained in 14 % yield was identified as the dihydrate of the title compound.

- NMR (400 MHz, DMSO-d₆) δ 7.35 (m, 5 H, P/zCH₂O-), 6.10 - 6.42 (m, 4 H , vinylic), 5.48 (q, 1 H, vinylic), 4.95 (m, 2 H, PhCH₂O-), 3.32 (s, 3 Η, -OC ₃), 3.18 (s, 3 Η, -OCH ), 3.05 (s, 3 Η, -OCHj); IR (KBr, cπr^l): 3420, 2920, 1730, 1645, 1445; MS (neg. FAB): 1003.4 [Ml^", 652, 590, 349; Anal. Calcd. for C53Η83NO13. 2 H₂O: C 66.98 %, H 8.56 , N 1.39 %; Found: C 66.53 %, H 8.51 %, N 1.70 %.

Preparation of substituted alkyl triflates.

To a solution of substituted alkanol (1.00 mmol) in dichloromethane (15.0 mL) in a round bottom flask (25 mL, flame dried) equipped with a magnetic stirrer at room temperature was added 2,6-di-t-butyl-4-methyl pyridine (0.60 g, 2.92 mmol). The reaction mixture was degassed, purged with nitrogen, and cooled to -78 °C. To the solution was added dropwise trifluoromethanesulfonic anhydride (0.170 mL, 0.282 g, 1.00 mmol) over a period of 5 minutes. The solution became a suspension. The reaction was stirred at 0 °C for 30 minutes, then wanned up to room temperature. TLC analysis showed completion of the reaction.

General Procedure for the Preparation of 42-substituted Derivatives of Rapamycin via Nucleophilic Substitution.

To the solution of substituted alkyl-triflate prepared from 1 mmol of alkanol was added at room temperature (unless otherwise noted) rapamycin (914 mg, 1 mmol). The mixture was stirred between 4 and 120 h (monitoring of the extent of the reaction was carried out by TLC). When desired conversion has been achieved the reaction was quenched with saturated aqueous NaHCO3, and the organic and aqueous layers were separated. The aqueous layer was extracted three times with ethyl acetate. The organic layers were combined, washed with brine, and dried over sodium sulfate. The solution was filtered and concentrated in vacuo to afford a product. Pure products were isolated by HPLC (normal phase- Dynamax 2" silica column, eluent EtOAc: hexane, 20 mL/min; reversed phase -Dynamax 2" Ci8 column, eluent MeCN: water, 20 mL/min. Spectroscopic analyses were used to confirm the structures.

Example 2.

42-O-(4-chlorobutyl)rapamycin

Method A. Alkanol used: 108 mg (1 mmol) of l-chloro-butane-4-ol. Reaction time: 72 h at 25 °C. Separation technique employed: 2" Dynamax silica column, eluent 40% EtOAc: hexane, 20 mL/min. Yield of product: 180 mg (18 %).

Spectral data follows: 1H NMR (400 MHz, DMSO-d6 δ 6.08-6.45 (m, 4 H), 3.4-3.7

( , 8 H), 3.15 (s, 3H), 3.04 (s, 3 H). IR (KBr, cm^"1) 3420, 2930, 1715, 1650, 1620, 1460. MS (neg. FAB) 1003.4 [Ml^", 590.2, 41 1.2. Anal, calcd. for

C55H86NO1 CI: C 65.75 %, H 8.63 %, N 1.39 %; Found: C 65.14 %, H 8.64 %,

N 1.18 %.

Example 3.

42-O-(2-[2-(2-methoxy ethoxy)-ethoxy]-ethyl)-rapamycin

Method A. Alkanol used: 165 mg (1 mmol) of triethylenegycol monomethyl ether. Reaction time: 120 h at 10 °C. Separation technique employed: 2" Dynamax silica column, eluent 70% EtOAc: hexane, 20 mL/min. Yield of product: 210 mg (19 %).

Spectral data follows: ^]H NMR (400 MHz, DMSO-d6 δ 6.05-6.45 (m, 4 H), 3.4-

3.65 (m, 12 H), 3.23 (s, 3 H), 3.14 (s, 3 H), 3.04 (s, 3 H). IR (KBr, cm^{" 1} ) 3420,

2925, 1720, 1645, 1450; MS (neg. FAB) 1059.4 fM]^", 590.2, 467.2. Anal. Calcd. for C58H93NO16. H2O C 64.60 %, H 8.81 %, N 1.28 %; Found: C 63.96 %, H

8.64 %, N 1.15 %. Pharmacology

Immunosuppressive activity was evaluated in an in vitro standard

5 pharmacological test procedure to measure lymphocyte proliferation (LAF), in an in vivo procedure to evaluate the survival time of a pinch skin graft, and in an in vivo procedure to determine inhibition of T-cell mediated inflammatory response (adjuvant arthritis).

The comitogen-induced thymocyte proliferation procedure (LAF) was used as K) an in vitro measure of the immunosuppressive effects of representative compounds. Briefly, cells from the thymus of noπnal 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. 15 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 μM.

³H-controI thymus cells - H³-rapamycin-treated thymus cells 20 ³H-control thymus cells - H³-test compound-treated cells

The results for the rapamycin analog (ICsnanalog) and rapamycin (ICsrjrapa) as well as the ratio of the ID50S of rapamycin to the analog (R/A) are given in the table below. A ratio less than 1.0 means the analog is less potent than rapamycin.

25 The 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. 28:385- 402, (1951). Briefly, a pinch skin graft from the donor is grafted on the dorsum of the recipient as a ho ograft, and an autograft is used as control in the same region. The

30 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 (MST -number of days + S.D.) of the drug treatment group is compared

35 with the control group. Rapamycin treatment provides a mean graft survival (MST) of 12.0± 1.7 days. The in vivo adjuvant arthritis standard pharmacological test procedure measures the ability of test compounds to prevent immune mediated inflammation and inhibit or treat rheumatoid arthritis. The following briefly describes the test procedure used. A group of rats (male inbred Wistar Lewis rats) are pre-rreated with the compound to be tested (lh prior to antigen) and then injected with Freund's Complete Adjuvant (FCA) in the right hind paw to induce arthritis. The rats are then orally dosed on a Monday, Wednesday, Friday schedule from day 0-14 for a total of 7 doses. Both hind paws are measured on days 16, 23 and 30. The difference in paw volume (mL) from day 16 to day 0 is determined and a percent change from control is obtained. The left hind paw (uninjected paw) inflammation is caused by T-cell mediated inflammation and is recorded as percent change from control. The right hind paw inflammation, on the other hand, is caused by non-specific inflammation. Compounds were tested at a dose of 2 g/kg. The results are expressed as the percent change in the uninjected paw at day 16 versus control; the more negative the percent change, the more potent the compound. Rapamycin provides between -70% and -90% change versus control, indicating that rapamycin treated rats have between 70-90% less immune-induced inflammation than control rats.

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

Table 1 : Summary of Pharmacological Test Results

Evaluation of Immunosuppressive Activity

LAF Skin Graft Adjuvant Arthritis

% Change

2 2.4 0.00

3 1.47 1.1 0.75 9.8 0.8 -88 3.10 0.8 0.26 9.7 0.5

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 test procedure indicate suppression of T cell proliferation. As transplanted pinch skin grafts are typically rejected with 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. The reduction of inflammatory joint swelling in the adjuvant rat model demonstrates their utility in the treatment of inflammatory diseases. Inhibition of growth of human tumors in vitro by rapamycin analogs

The compound of Example 3 was shown to inhibit in submicromolar concentrations breast (T47D), colon (MIP 101), and ovarian (A 2780S) cancer cell lines according to the following assay procedure:

Human tumor cell lines were plated in 96-well plates (250 μL/well, 1-6 x 10⁴ cells/mL) in RPMI 1640 medium, containing 5% FBS (Fetal Bovine Serum). Twenty- four hours after plating, drugs were added at five log concentrations (0.01 -100 μg mL). After 48 hours exposure to drugs, cells were fixed with trichloroacteic acid, and stained with Sulforhodamine B. After washing with trichloroacetic acid, bound dye was solubilized in 10 mM Tris base and Optical Density was determined using a plate reader. Under conditions of the assay, the optical density is proportional to the number of cells in the well. IC50S (concentrations causing 50% inhibition of cell growth) were deteπnined from the growth inhibition plots. The assay is described in details by Philip Skehan et al., J. National Cancer Institute 82, 1107-11 12, 1990.

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 ellitus, yasthenia gravis, and multiple sclerosis; and diseases of inflammation such as, psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease and pulmonary inflammation such as asthma, chronic obstructive pulmonary disease, emphysema, bronchitis and the like; proliferative diseases such as restenosis following angioplasty procedures, and fungal infections. The compounds are also expected to be useful in the treatment of breast, colon, or ovarian cancers in humans.

Pharmaceutical Composition

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 and the active compound shall be a therapeutical ly effective amount.

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 can 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 propellant.

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. The formulated compound can further be administered intranasally through insufflation of a powder formulation, rectally or vaginally via suppositories, and topically or transdermally. Furthermore, the formulated invention compound can be administered alone or in combination with one or more addidional immunoregluatory agents such as a corticosteroid, cyclophosphamide, rapainyucin, cyclosporin A, FK-506, OKT-3 or ATG as established by Stepkowski, Transplantation Proceedings 23: 507 (1991).

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 doses 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 neatment must be subjectively determined by the attending physician.

Claims

What is claimed is:

1. A compound having the formula:

wherein W and Y are OR¹ and X and Z together form a bond or W and X are OR² and Y and Z together form a bond, wherein:

R¹ is selected from -(CH2)_n-Ar where Ar is not phenyl, -(CH₂CH2O)_nCH3 where n is not 1 , -CH2CH2CH₂O(CH2CH2O)_m-CH3, -(CH₂)n-CH2CH(OR )CH₂OR⁴ where R³ and R⁴ are H, C]-Cι₀ alkyl, or R³ and R⁴ together tire ethylene, methylene or dimethylmethylene; -CH2(CH₂)_n-OR³ where R³ is not H, Cj-Cio alkyl, or C(O)C]-Cι₀ alkyl; and -CH₂(CH₂)_n-X where X is F, Cl, Br or I;

R² is selected from H, Cj-Cio alkyl, Ar(CH₂)_n-, C3-C10 alkenyl, -(CH₂CH₂O)_nCH3, -CH2CH₂CH₂O(CH2CH2O)_m-CH3, -CH₂(CH₂)n-OR³, -CH₂(CH₂)_n-X where X is F, Cl, Br or I; and -(CH₂)_n-CH₂CH(OR⁵)CH₂OR⁶ where R5 and R6 are selected independently from H, C]-Cιo alkyl, -(CH₂)_n-Ar, -CONH(CH₂)_n-Ar or COC(CH3)₂-(CH₂)n-Ar, -COR? and -CO₂R⁷, where R⁷ is C]-C₆ alkyl, C2-C6 alkenyl, or Ar; n = 1 -10 independently; m = 1-5 independently; and

Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;

1, 2, 3-triazolyI and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.

2. A compound according to claim 1 which is 29-O-benzylrapamycin.

3. A compound according to claim 1 which is 42-O-(4-chlorobutyl)rapamycin.

4. A compound according to claim 1 which is 42-O-(2-[2-(2-methoxyethoxy)- ethoxy]ethyl)rapamycin

5. A method of treating transplantation rejection or host vs. graft disease in a mammal by administering thereto an immunosuppressing effective amount of a compound having the formula:

R¹ is selected from -(CH2)_n-Ar where Ar is not phenyl, -(CH₂CH₂O)_nCH3 where n is not 1 , -CH₂CH2CH2θ(CH₂CH2θ)_m-CH3,

-(CH₂)_n-CH₂CH(OR³)CH₂OR⁴ where R³ and R⁴ are H, Ci-Cm alkyl, or R³ and R⁴ together are ethylene, methylene or dimethylmethylene; -CH₂(CH₂)_n-OR³ where R³ is not H, Cj-Cio alkyl, or C(O)Cι-Cιo alkyl; and -CH₂(CH₂)n-X where X is F, Cl, Br or I;

R² is selected from H, C I -C ]Q alkyl, Ar(CH₂)_n-, C3-C_{] 0} alkenyl, -(CH2CH₂O)_nCH3, -CH₂CH2CH2θ(CH2CH2θ)_m-CH₃, -CH₂(CH₂)n-OR³, -CH₂(CH₂)_n-X where X is F, Cl, Br or I; and -(CH₂)n-CH₂CH(OR5)CH2OR⁶ where R⁵ and R⁶ are selected independently from H, Cj-Cio alkyl, -(CH2)_n-Ar, -CONH(CH₂)n-Ar or COC(CH3)2-(CH₂)_n-Ar, -COR⁷ and -CO₂R⁷, where R⁷ is C]-C₆ alkyl, C2-C6 alkenyl, or Ar; n= 1-10 independently; m= 1-5 independently; and

1 , 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.

6. A method of treating asthma in a mammal by administering thereto an asthma inhibiting effective amount of a compound having the formula

wherein W and Y are OR' and X and Z together form a bond or W and X are OR² and Y and Z together form a bond, wherein:

R¹ is selected from -(CH2)_n-Ar where Ar is not phenyl,

-(CH₂CH₂O)nCH3 where n is not 1,

-CH2CH₂CH2θ(CH2CH2θ)_n CH3,

-(CH₂)_n-CH₂CH(OR )CH₂OR⁴ where R³ and R⁴ are H, Ci-Cio alkyl, or R³ and R⁴ together are ethylene, methylene or dimethylmethylene; -CH₂(CH2)n-OR³ where R³ is not H, Cj-Cio alkyl, or C(O)C]-Cιo alkyl; and -CH₂(CH2)_n-X where X is F, Cl, Br or I;

R² is selected from H, Ci-Cio alkyl, Ar(CH₂)_n-, C -CH) alkenyl, -(CH CH2θ)_nCH3, -CH₂CH₂CH2O(CH2CH₂O)_m-CH3, -CH2(CH₂)_n-OR³, -CH₂(CH₂)n-X where X is F, Cl, Br or I; and -(CH₂)n-CH2CH(OR5)CH₂OR⁶ where R⁵ and R⁶ are selected independently from H, C₁-C₁0 alkyl, -(CH )_n-Ar, -CONH(CH₂)_n-Ar or COC(CH3)2-(CH₂)_n-Ar, -COR⁷ and -CO R⁷, where R⁷ is C1-C6 alkyl, C2-C6 alkenyl, or Ar; n= 1-10 independently; m= 1-5 independently; and

7. A method of treating rheumatoid arthritis in a mammal by administering thereto an arthritis inhibiting effective amount of a compound having the formula:

R- is selected from -(CH2)_n-Ar where Ar is not phenyl, -(CH₂CH2O)„CH3 where n is not 1, -CH2CH₂CH2θ(CH2CH2θ)m-CH , -(CH₂)_n-CH₂CH(OR³)CH2θR⁴ where R³ and R⁴ are H, Ci-Cin alkyl, or R³ and R⁴ together are ethylene, methylene or dimethylmethylene; -CH₂(CH₂)_n-OR³ where R³ is not H, Ci-Cjo alkyl, or C(O)Cι-Cκ) alkyl; and -CH2(CH₂)n-X where X is F, Cl, Br or I;

R² is selected from H, Ci-Cin alkyl, Ar(CH₂)_n-, C3-C10 alkenyl, -(CH2CH₂O)_nCH3, -CH₂CH2CH2O(CH2CH₂O)nrCH3, -CH₂(CH₂)n-OR³, -CH₂(CH )_n-X where X is F, Cl, Br or I; and -(CH₂)_n-CH₂CH(OR^<5)CH₂OR⁶ where R⁵ and R⁶ are selected independently from H, Cj-Cio alkyl, -(CH )_n-Ar, -CONH(CH₂)_n-Ar or COC(CH3)2-(CH₂)_n-Ar, -COR⁷ and -CO₂R⁷, where R⁷ is Cj-C₆ alkyl, C2-C6 alkenyl, or Ar; n= 1-10 independently; m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl; 1, 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.

8. A method of treating fungal infections in a mammal by administering thereto an antifungal effective amount of a compound having the formula:

wherein W and Y are OR' and X and Z together foπn a bond or W and X arc OR² and Y and Z together form a bond, wherein:

R- is selected from -(CT r where Ar is not phenyl, -(CH2CH O)_nCH₃ where n is not 1 , -CH2CH2CH2θ(CH₂CH2θ) -CH3, -(CH₂)_n-CH₂CH(OR³)CH₂OR⁴ where R³ and R⁴ are H, C]-C₁₀ alkyl, or R³ and R⁴ together are ethylene, methylene or dimethylmethylene; -CH2(CH₂)_ιrOR³ where R³ is not H, Ci-Cio alkyl, or C(O)Cι-Cιo alkyl; and -CH₂(CH₂)_n-X where X is F, Cl, Br or I;

R² is selected from H, Cι-Cιo alkyl, Ar(CH₂)_n-, C₃-C₁₍) alkenyl, -(CH2CH₂O)_nCH₃, -CH2CH₂CH2O(CH2CH₂O)_m-CH3, -CH₂(CH₂)_n-OR³, -CH₂(CH2)_n-X where X is F, Cl, Br or I; and -(CH₂)„-CH₂CH(OR⁵)CH₂OR⁶ where R⁵ and R⁶ are selected independently from H, Cj-Cio alkyl, -(CH₂)_n-Ar, -CONH(CH₂)_n-Ar or COC(CH3)₂-(CH2)n-Ar, -COR⁷ and -CO2R⁷, where R⁷ is -Cό alkyl, C2-Q, alkenyl, or Ar; n= 1-10 independently; m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl; 1 , 2, 3-triazolyI and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.

9. A method of inhibiting restenosis in a mammal by administering thereto a restenosis inhibiting effective amount of a compound having the formula:

R¹ is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH₂O)_nCH3 where n is not 1 , -CH2CH2CH2θ(CH₂CH2θ)„_rCH3,

-(CH₂)_n-CH₂CH(OR³)CH2θR⁴ where R³ and R⁴ are H, CJ-CJO alkyl, or R³ and R⁴ together are ethylene, methylene or dimethylmethylene; -CH₂(CH2)_n-OR³ where R³ is not H, C1-C10 alkyl, or C(O)Cι-Cιo alkyl; and -CH2(CH2)n-X where X is F, Cl, Br or I; R² is selected from H, Ci-Cio alkyl, Ar(CH₂)_n-, C3-C10 alkenyl, -(CH₂CH₂O)nCH3, -CH2CH2CH2θ(CH2CH2θ)_m-CH3, -CH₂(CH₂)n-OR³, -CH₂(CH₂)_n-X where X is F, Cl, Br or I; and -(CH₂)_n-CH₂CH(OR⁵)CH₂OR⁶ where R⁵ and R⁶ are selected independently from H, C₁-C10 alkyl, -(CH₂)_n-Ar, -CONH(CH₂)_n-Ar or COC(CH₃)₂-(CH2)n-Ar, -COR⁷ and -CO₂R⁷, where R⁷ is Cι-C₆ alkyl, C2-C6 alkenyl, or Ar; n= 1-10 independently; m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;

10. A method of treating breast cancer, colon cancer, or ovarian cancer which comprises administration to a person having such a cancer of a cancer-inhibiting amount of a compound having the formula:

R¹ is selected from -(CH2)_n-Ar where Ar is not phenyl, -(CH₂CH₂O)_nCH3 where n is not 1, -CH₂CH2CH2θ(CH2CH₂O)_m-CH3,

-(CH₂)_n-CH₂CH(OR³)CH₂OR⁴ where R³ and R⁴ are H, C_\-C_]0 alkyl, or R³ and R⁴ together are ethylene, methylene or dimethylmethylene; -CH₂(CH₂)_n-OR³ where R³ is not H, C]-CJO alkyl, or C(O)C]-Cιo alkyl; and -CH₂(CH₂)_n-X where X is F, Cl, Br or I;

R² is selected from H, Cj-Cio alkyl, Ar(CH₂)_n-, C3-C10 alkenyl, -(CH₂CH2θ)_nCH3, -CH2CH2CH2O(CH₂CH2O)m-CH3, -CH₂(CH₂)_n-OR³, -CH₂(CH₂)_n-X where X is F, Cl, Br or I; and -(CH₂)_n-CH₂CH(OR⁵)CH₂OR⁶ where R⁵ and R⁶ are selected independently from H, C]-CIQ alkyl, -(CH₂)n- Ar, -CONH(CH₂)_n-Ar or COC(CH₃)2-(CH₂)n-Ar, -COR⁷ and -CO₂R⁷, where R⁷ is Cι-C₆ alkyl, C2- alkenyl, or Ar; n= 1-10 independently; m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;

1 1. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of the formula:

R¹ is selected from -(CH2)_n-Ar where Ar is not phenyl,

-(CH₂CH₂O)_nCH3 where n is not 1,

-CH₂CH₂CH₂O(CH₂CH2O)_m-CH3,

-(CH₂)n-CH₂CH(OR³)CH2θR⁴ where R³ and R⁴ are H, CJ-CJO alkyl, or R³ and R⁴ together are ethylene, methylene or dimethylmethylene; -CH2(CH₂)_n-OR³ where R³ is not H, Ci-Cio alkyl, or C(O)Cι-Cιo alkyl; and -CH₂(CH₂)n-X where X is F, Cl, Br or 1;

R² is selected from H, C|-Cιo alkyl, Ar(CH₂)_n-, C3-C10 alkenyl, -(CH₂CH₂O)nCH3, -CH2CH2CH2θ(CH2CH₂O)_m-CH3, -CH₂(CH₂)n-OR³, -CH₂(CH₂)_n-X where X is F, Cl, Br or l; and -(CH₂)_n-CH2CH(OR⁵)CH₂OR⁶ where R⁵ and R⁶ are selected independently from H, CI-CJO alkyl, -(CH₂)_n-Ar, -CONH(CH₂)_n-Ar or COC(CH₃)2-(CH₂)_n-Ar, -COR⁷ and -CO R⁷, where R⁷ is C1-C6 alkyl, C₂-C6 alkenyl, or Ar; n= 1 - 10 independently; m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;

1, 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.