MXPA97003893A

MXPA97003893A - Un-oxido-impaired esters of rapamycin and its uses as a medicine

Info

Publication number: MXPA97003893A
Application number: MXPA/A/1997/003893A
Authority: MX
Inventors: Alan Schiehser Guy; Christy Nelson Frances
Original assignee: American Home Products Corporation
Priority date: 1994-11-28
Filing date: 1997-05-27
Publication date: 1997-08-01

Abstract

A compound of structure (I) is described, wherein R and R1 are each independently, a), b), or hydrogen: R2 and R3 are each, independently, alkyl, arylalkyl, or R2 and R3 can be taken together to form a cycloalkyl ring, R4 is a heterocyclic N-oxide radical, which may be optionally substituted, R5 is alkyl or arialkyl, R6 and R7 are taken together to form a saturated N-alkyl-heterocyclic N-oxide, which may be be optionally substituted, k = 0-1, m = 0-1, n = 1-6, with the proviso that R and R1 are not both hydrogen, which is useful as an immunosuppressant, anti-inflammatory, antifungal, antiproliferative and antitumor agent

Description

N-OXIDE - IMPAIRED THEMES OF RAPAMYCIN AND ITS USES AS MEDICINES BACKGROUND OF THE INVENTION This invention relates to the hindered N-oxide-esters of rapa icin and to a method for using them for the induction of immunosuppression, and in the treatment of rejection to transplantation, graft versus host disease, autoimmune diseases, inflammation diseases, leukemia. adult T-cell lymphoma, solid tumors, fungal infections, and hyperproliferative vascular disorders. Rapamycin is a macrocyclic trienic antibiotic produced by Streptoinyces hygroscopicus, which was found to have antifungal activity, particularly against Candida albi cans, either in vi tro e 'in vi vo [C. Vezina et al., J. Antibiot. 28, 721 (1975); S.N. Sehgal et al., J. Antibiot. 28, 727 (1975); HE HAS. Baker et al., J. Antibiot. 31, 539 (1978); U.S. Patent No. 3,929,992; and North American patent No. 3,993,749] REF: 24725 Rapamycin alone (U.S. Patent No. 4,885,171) or in combination with picibanil (U.S. Patent No. 4,401,653) has been shown to have antitumor activity. R. Martel et al [Can. J. Physiol. Pharmacol. 55, 48 (1977)] reported that rapamycin is effective in the experimental model of allergic encephalomyelitis, 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 described in FASEB 3, 3411 (1989).

Cyclosporin A and FK-506, other macrocyclic molecules, have also been shown to be effective as immunosuppressive agents, therefore useful in the prevention of rejection to transplantation [FASEB 3, 3411 (1989); FASEB 3, 5256 (1989); R. Y. Calne et al., Lancet 1183 (1978); and U.S. Patent No. 5,100,899]. It has also been shown that rapamycin is useful in the prevention or treatment of systemic lupus erythematosus [U.S. Patent No. 5,078,999], pulmonary inflammation [U.S. Patent No. 5,080,899], insulin-dependent diabetes mellitus [Fifth Int. Conf. Inflamm . Res. Assoc. 121 (Abstract), (1990)], smooth muscle cell proliferation and thickening of the intima after vascular damage [Morris, RJ Heart Lung Transplant 11 (pt.2): 197 (1992)], leukemia / cell lymphoma Adult T [European Patent Application No. 525,960 Al], and ocular inflammation [European Patent Application 532, 862 Al]. The mono- and diacylated derivatives of rapamycin (esterified at positions 28 and 43) have been shown to be useful as antimicrobial agents (U.S. Patent No. 4,316,885), and used to make water-soluble aminoacylated prodrugs of rapamycin (U.S. Pat. 4,650,803). Recently, the numbering convention for rapamycin has been changed; therefore, according to the Chemical Abstacts nomenclature, the esters described above could be in positions 31 and 42.

DESCRIPTION OF THE INVENTION This invention provides rapamycin derivatives which are useful as immunosuppressive, anti-inflammatory, antifungal, antiproliferative and antitumor agents, which have the structure wherein R and R: are each, independently, or hydrogen; R2 and R3 are each, independently, alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, or R2 and R3 can be taken together to form a cycloalkyl ring of 3 to 8 carbon atoms; R4 is a heterocyclic N-oxide radical of 5 to 12 carbon atoms, which may be optionally mono-, di-, or tri- substituted with a group selected from alkyl of 1 to 6 carbon atoms, arylalkyl of 7 to 10. carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7. carbon atoms, trifluoromethyl, trifluoromethoxy, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 carbon atoms, -S03H, -P03H2, and -C02H; R5 is alkyl of 1 to 6 carbon atoms or arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms; R6 and R7 are taken together to form an N- (C 1-6 -alkyl) -N-oxide-heterocyclic ring of 5 to 8 atoms, which may be optionally mono- or tri- substituted. by a group selected from alkyl of 1 to 6 carbon atoms, aroyl of 3 to 11 carbon atoms, and perfluoroalkyl of 1 to 6 carbon atoms; k-0-1; m = 0-1; n-1 - 6; with the proviso that R and R1 are not both hydrogen. It is preferred that the heterocyclic N-oxide radical defined in R 4 be an unsaturated or partially saturated heterocyclic N-oxide radical of 5 to 12 carbon atoms, having one ring or two fused rings. Preferred heterocyclic N-oxide radicals include the unsaturated heterocyclic N-oxide radicals such as 1-methyl-pyrazolyl-2-N-oxide, imidazolyl-3-N-oxide, 2- or 3-N-oxido of 1, 2, 3-triazolyl, 2- or 4-N-oxide of 1,2,4-triazolyl, N-oxide of 1, 2, 5, -oxadiazolyl, N-oxide of 1, 2, 3, 5-oxatriazolyl, pyridinyl N-oxide, pyridazinyl N-oxide, pyrimidinyl N-oxide, pyrazinyl N-oxide, 1,3,5-triazinyl N-oxide, N-oxide 1, 2, 4-triazinyl, 1,2,3-triazinyl N-oxide, 1,2,4-diazepinyl N-oxide, 2-isobenzazolyl N-oxide, 1,5-pyrindinyl N-oxide , Benzopyrazolyl N-oxide, Benzisoxazolyl N-oxide, benzoxazolyl N-oxide, quinolinyl N-oxide, isoquinolinyl N-oxide, cinnolinyl N-oxide, quinazolinyl N-oxide, naphthyridinyl N-oxide, pyrido-N-oxide [3,4 -b] pyridinyl, pyrido [4, 3-b] pyridinyl N-oxide, pyrido [2, 3-b] pyridinyl N-oxide, 1,4,2-benzoxazinyl N-oxide, N-2-oxide , 3, 1-benzoxazinyl, carbazolyl N-oxide, purinyl N-oxide, and partially saturated heterocyclic N-oxide radicals selected from the above list. All preferred heterocyclic N-oxide radicals contain at least one double bond. When the heterocyclic N-oxide radical is partially saturated, one or more of the olefins in the unsaturated ring system is saturated; the partially saturated heterocyclic N-oxide radical contains at least one double bond. The side chain - (CH2) n- may be coupled at any position of the heterocyclic N-oxide radical containing a carbon or nitrogen atom capable of forming a bond with the side chain - (CH2) n-. The most preferred heterocyclic N-oxide radicals are pyridinyl N-oxide, pyrazinyl N-oxide, triazinyl N-oxide, pyrimidinyl N-oxide, pyridazinyl N-oxide, imidazolyl N-oxide, pyrazolyl N-oxide , Quinolinyl N-oxide, and isoquinolinyl N-oxide. The pyridinyl N-oxide is the most preferred heterocyclic N-oxide radical. It is preferred that saturated N-alkyl heterocyclic of 1 to 6 carbon atoms, N-oxide of 5 to 8 ring atoms as defined by R6 and R7, are an N-oxide of N- (alkyl of 1 to 6 carbon atoms) -piperidine, N-oxide of N- (alkyl of 1 to 6 carbon atoms) -morpholine, N-oxide of N- (alkyl of 1 to 6 carbon atoms) -piperazine, N-oxide of N- (C 1-6 alkyl) -pyrazolidine, N- (C 1-6 alkyl) -imidazolidine N-oxide, or N-oxide N- (C 1-6 alkyl) -pyrrolidine. Methyl is the preferred alkyl group. Aroyl is defined as the radical Ar-CO-, where Ar is an aryl radical. The term "aryl" as a group or part of a group such as arayl or arylalkyl includes any carbocyclic aromatic group of 6 to 10 carbon atoms or heteroaromatic group of 5 to 10 ring atoms, of which up to 3 ring atoms they are heteroatoms selected from the group consisting of oxygen, nitrogen- and sulfur. When the aryl group is substituted, the examples of substituents are one or more, the same or different from the following: alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, C 1 -C 6 alkoxy, cyano, halo, hydroxyl, hydroxy-alkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 carbon atoms, and dialkylamino-alkyl of 3 to 12 carbon atoms, nitro, carbalkoxy from 2 to 7 carbon atoms, trifluoromethyl, amino, mono- or di-alkylamino of 1 to 6 carbon atoms per alkyl, aminocarbonyl, alkylthio group of 1 to 6 carbon atoms, -S03H, -P03H and -C03H. The aryl group can be mono- or bicyclic. It is preferred that the aryl portion of the arylalkyl group and the aroyl group be a phenyl, naphthyl, pyridinyl, quinolinyl, isoquinolinyl, furanyl, benzofuranyl, benzodioxyl, benzoxazolyl, benzoisoxazolyl, indolyl, isoxazolyl, pyrimidinyl, pyrazinyl, benzopyranyl, or benzylol ideolyl group, which may optionally be mono-, di-, or tri-substituted with a group selected from alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7 carbon atoms, trifluoromethyl, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 carbon atoms, -S03H, -P03H2, and -C02H. It is more preferred that the aryl portion is a phenyl group which may be optionally substituted as described above. The term alkyl of 1 to 6 carbon atoms includes the straight chain as well as the branched carbon chains. Examples of alkyl as a group or part of a group, for example, arylalkyl, alkoxy or alkanoyl (alkylcarbonyl) are straight or branched chains of 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms, for example, methyl , ethyl, propyl, isopropyl and n-butyl. Of the compounds of this invention, the preferred members are those in which R1 is hydrogen; those in which R1 is hydrogen and R is When R is JL -? CH »j¡ * =" «> and R1 is. hydrogen, the preferred compounds are those in which R4 is pyridinyl N-oxide, pyrazinyl N-oxide, triazinyl N-oxide, pyrimidinyl N-oxide, pyridazinyl N-oxide, imidazolyl N-oxide, N- pyrazolyl oxide, quinolinyl N-oxide, or isoquinolinyl N-oxide; those in which R 4 is pyridinyl N-oxide; those in which R 4 is pyridinyl N-oxide, and k = 0; those in which R 4 is pyridinyl N-oxide, k = 0, and R 2 and R 3 are alkyl of 1 to 6 carbon atoms or are taken together to form a cycloalkyl ring of 3 to 8 carbon atoms. This invention provides a process for the preparation of the rapamycin compounds of this invention. In particular, this invention provides a process for the preparation of N-oxide hindered esters of rapamycin, including those of the formula I as defined above which comprises: a) the acylation of rapamycin or a functional derivative thereof with an acylating agent or b) the sequential acylation of rapamycin or a functional derivative thereof with two acylating agents said acylating agents are selected from the acids of the formula (IIa) (Hb) wherein n, k, m and R2-R7 are as defined above, or a reactive derivative thereof, if desired protecting the 42 position of rapamycin with an appropriate protecting group and eliminating the same as is required The reaction can be carried out in the presence of a coupling agent, such as a suitably substituted carbodiimide coupling reagent. The above-mentioned compounds of this invention can also be prepared by acylation using reactive derivatives of the acid of formula Ia and Ilb, such as an anhydride, a mixed anhydride, or an acid halide such as the chloride.

The compounds that contain the ester group JL ¡? CCHH j), - ^ ° H = > "k7 at positions 42- or 31, 42, can be prepared by treatment of a suitably substituted carboxylic acid, with a hindered base such as LDA, followed by alkylation with a heterocycle containing haloalkyl-nitrogen. containing nitrogen can be oxidized to the corresponding N-oxide using an oxidation agent such as m-chloroperbenzoic acid (MCPBA) The resulting alkylated acid can then be activated as a mixed anhydride, with an acylating group such as 2, 4 chloride, 6-trichlorobenzoyl The treatment of rapamycin with the mixed anhydride under slightly basic condition provides the desired compounds.The mixtures of the 42- and 31,42-esters can be separated by chromatography.This scheme is described below. and the haloalkyl-heterocyclics are either commercially available or can be prepared by the standard procedures of the to literature.

R4 > pyridyl MCPBA / CH2CI2 12. p «rp¿am.ciir DMAP" * - Compounds that contain the ester group in positions 42- or 31, 42 can be prepared analogously. The mixtures of the -esters at positions 42- and 31, 42 can be separated by chromatography. The esters in theposition31 of this invention can be prepared by protecting the 42-alcohol of rapamycin with a protecting group, such as with a tert-butyl-dimethylsilyl group, followed by esterification of the 31-position by the procedures described above. The preparation of the 42-silyl ethers of rapamycin is described in U.S. Patent No. Bl 5,120,842, which is incorporated by reference herein. Removal of the protecting group provides the 31-esterified compounds. In the case of the tert-butyl-dimethylsilyl protecting group, the deprotection can be achieved under mildly acidic conditions, such as acetic acid / water / tetrahydrofuran. The deprotection process is described in Example 15 of U.S. Patent No. 5, 118,678, which is incorporated by reference herein. Having the 31-position and the deprotected position 42 esterified, the 42-position can be esterified using a different acylating agent that was reacted with the 31-alcohol, to give the compounds having different esters at the 31 and 42 positions. Alternatively, 42-esterified compounds, prepared as described above, can be reacted with a different acylating agent to provide the compounds having different esters at positions 31 and 42. This invention also covers the hindered analogs of functional derivatives of rapamycin such as, but not limited to, 29-demethoxyrapamycin, [U.S. Patent No. 4,375,464, 32-demethoxyrapamycin under the CA nomenclature]; rapamycin derivatives in which the double bonds at positions 1-, 3-, and / or 5 have been reduced [U.S. Patent No. 5,023,262]; 29-desmethylrapamycin [U.S. Patent No. 5,093,339, 32-demethylrapamycin under the nomenclature of C.A.]; 7, 29-bisdesmethylrapamycin [U.S. Patent No. 5,093,338, 7, 32-desmethylrapamycin under the nomenclature of C.A.]; and 15-hydroxyrapamycin [U.S. Patent No. 5,102,876]. This invention also covers hindered esters at position 31 of 42-oxorapamycin [U.S. Patent No. 5,023,263]. The descriptions in the aforementioned US patents are incorporated by reference herein. The immunosuppressant activity for the representative compounds of this invention was evaluated in a standard pharmacological test procedure, in vi tro to measure the proliferation of lymphocytes (LAF) and in a standard in vitro pharmacological test procedure, which measures the immunosuppressive activity of the tested compound, as well as the ability of the compound tested to inhibit or treat rejection of the transplant. The procedures for these standard pharmacological test procedures are provided below. The procedure of proliferation of thymocyte-induced comitogen (LAF) was used as a measure of the immunosuppressive effects of the 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 varying concentrations of rapamycin, cyclosporin A, or the test compound. The cells are harvested and the incorporated radioactivity is determined. Inhibition of lymphoproliferation is evaluated as the percentage change in counts per minute from controls not treated with the drug. For each compound evaluated, rapamycin was also evaluated for comparison purposes. An IC50 value was obtained for each test compound, as well as for rapamycin. When evaluated as a comparator for the representative compounds of this invention, rapamycin had an IC50 of 2.4 nM. The results obtained are given as an IC50.

The representative compounds of this invention were also evaluated in an in vivo test procedure designed to determine the survival time of the skin graft by puncture from male BALB / c donors transplanted into male C3H (H-2K) vessels. The method is adapted from Billingham R.E. and Medawar P.B., J. Exp. Biol. 28: 385-402, (1951). In summary, a skin graft by puncture from the donor was grafted onto the back of the recipient as a halograft, and an isograft was used as a control in the same region. The containers were treated either with varying concentrations of the test compounds intraperitoneally or orally. Rapamycin was used as a test control. Untreated containers serve as the rejection control. The graft was checked periodically on a daily basis and observations were recorded until the graft became dry and a blackened scab formed. This was considered as the day of rejection. The mean survival time of the graft (number of days ± S.D.) of the treatment group with the drug was compared with the control group. The following table shows the results that were obtained. The results are expressed as the average survival time in days. Skin grafts by puncture, untreated (control) are usually rejected within 6 to 7 days. The compounds were tested using a dose of 4 mg / kg administered intraperitoneally or using a dose of 40 mg / kg administered p.o. The results obtained in these standard pharmacological test procedures are provided after the procedure for the elaboration of the specific compounds that were tested. The results of these standard pharmacological test procedures demonstrate the immunosuppressive activity either in. vi tro and vi vi for the compounds of this invention. The results obtained in the LAF test procedure indicate the suppression of T cell proliferation, thereby demonstrating the immunosuppressive activity of the compounds of this invention.

Further demonstration of the utility of the compounds of this invention as immunosuppressive agents was shown by the results obtained in the standard skin graft pharmacological test procedure. Additionally, the results obtained in the skin graft test procedure further demonstrate the ability of the compounds of this invention to treat or inhibit rejection to transplantation. Based on the results of these standard pharmacological test procedures, the compounds are useful in the treatment or inhibition of rejection to transplantation such as kidney, heart, liver, lung, bone marrow, pancreas (islet cells), cornea, intestine. thin, and skin allografts, and heart valve xenografts; in the treatment or inhibition of autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, myasthenia gravis, and multiple sclerosis; and inflammation diseases such as psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease, and ophthalmic uveitis. Due to the profile of activity obtained, the compounds of this invention are also considered as possessing antitumor and anti-fungal activities, and antiproliferative activities. The compounds of this invention are therefore also useful in the treatment of solid tumors, adult T-cell leukemia / lymphoma, fungal infections and hyperproliferative vascular diseases such as restenosis and atherosclerosis.

When administered for the treatment or inhibition of the above disease states, the compounds of this invention can be administered to a mammal orally, parenterally, intranasally, intrabronchially, transdermally, topically, intravaginally or rectally. It is contemplated that when the compounds of this invention are used as an immunosuppressive or anti-inflammatory agent, these can be administered in conjunction with one or more other immunoregulatory agents. Such immunoregulatory agents include, but are not limited to azathioprine, corticosteroids, such as prednisone and methylprednisolone, cyclophosphamide, rapamycin, cyclosporin A, FK-506, OKT-3, and ATG. By combining the compounds of this invention with other drugs or such agents, to induce immunosuppression or treatment of inflammatory conditions, lesser amounts of each of the agents are required to achieve the desired effect. The basis for such combination therapy was established by StepkOwski, whose results showed that the use of a combination of rapamycin and cyclosporin A at subtherapeutic doses significantly prolonged the prolonged survival time of the cardiac halograft [Transplantation Proc. 23: 507 (1991)]. The compounds of this invention can be formulated pure or with a pharmaceutical carrier to a mammal in need thereof. The pharmaceutical carrier can be solid or liquid. When formulated orally, it has been found that 0.01% Tween 80 in PHOSAL PG-50 (phospholipid concentrate with 1,2-propylene glycol, A. Nattermann & amp;; Company GmbH) provides an acceptable oral formulation. A solid carrier may include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending, fillers, glidants, compression aids, binding agents or tablet disintegrators; This can also be an encapsulation material. In the form of 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 appropriate proportions, and compacted in the desired shape and size. The powders and tablets preferably contain 99% active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, low melting point waxes, and ion exchange resins. . Liquid carriers are used in the repair of solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid carrier may contain other suitable pharmaceutical derivatives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, savoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators.

Suitable examples of liquid carriers for oral and parenteral administration include water (which partially contains additives such as the above, for example, cellulose derivatives, preferably sodium carboxymethylcellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, for example glycols) and their derivatives, and oils (for example fractionated coconut oil and arachis oil). For parenteral administration, the carrier can also be an oily ester such as oleate. ethyl and isopropyl myristate. Sterile liquid carriers are useful in sterile liquid form compositions for parenteral administration. The liquid carrier for the pressurized compositions may be halogenated hydrocarbon or other pharmaceutically acceptable propellant. Liquid pharmaceutical compositions which are sterile solutions or suspensions may be used by, for example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously. The compound can also be administered orally either in the form of a liquid or solid composition. The compounds of this invention can be administered rectally in the form of a conventional suppository. For administration by intranasal or intrabronchial inhalation or insufflation, the compounds of this invention can be formulated in an aqueous or partially aqueous solution, which can then be used in the form of an aerosol. The compounds of this invention can also be administered transdermally through the use of a transdermal patch containing the active compound and a carrier which is inert to the active compound, which is non-toxic to the skin, and which allows agent distribution. for systemic absorption into the bloodstream via the skin. The carrier can take any number of forms such as creams and ointments, pastes, gels, and occluder devices. The creams and ointments can be liquid viscous or semi-solid emulsions, either oil-in-water or water-in-oil. Pastes comprised of absorptive powders dispersed in petroleum or hydrophilic petroleum containing the active ingredient, may also be appropriate. A variety of occluder devices can be used to release the active ingredient into the bloodstream, such as a semipermeable membrane that covers a reservoir containing the ingredient with or without a carrier, or a matrix containing the active ingredient. Other occluder devices are known in the literature. In addition, the compounds of this invention can be employed as a solution, cream, or lotion by formulating with pharmaceutically acceptable carriers containing 0.1-5%, preferably 2%, of the active ingredient, which can be administered to an affected area by mushrooms. The dosage requirements vary with the particular compositions employed, the route of administration, the severity of the symptoms presented in the particular subject being treated. Based on the results obtained in the standard pharmacological test procedures, the projected daily doses of the active compound could be 0.1 μg / kg - 100 mg / kg, preferably between 0.001 - 25 mg / kg, and more preferably between 0.01 - 5 mg / kg. The treatment will generally be initiated with small doses less than the optimum dose of the compound. After this, the dose is increased until it reaches the optimum effect under the circumstances; The precise doses for oral, parenteral, nasal, or intrabronchial administration will be determined by the doctor who administers based on the experience with the individual subject treated. Preferably, the pharmaceutical composition is a unit dose form, for example, as tablets or capsules. In such form, the composition is subdivided into unit doses containing appropriate amounts of the active ingredient; the unit dosage forms may be packaged compositions, for example, packaged powders, flasks, ampoules, pre-filled syringes or sacks containing liquids. The unit dosage form may be, for example, a capsule or tablet itself, or this may be the appropriate number of any such compositions in the packaged form. The following examples illustrate the preparation and biological amounts of the representative compounds of this invention.

Example 1 2, 2-dimethyl-3- (3-pyridinyl) propionic acid Sodium hydride (4.38 g, 110 mmol, 60% dispersion, washed twice with hexanes and dried under nitrogen atmosphere) in tetrahydrbfuran (140 mL) was suspended. To this suspension was added diisopropylamine (15.4 ml, 110 mmol). Isobutyric acid (9.27 ml, 100 mmol) was slowly added dropwise. The resulting thick white suspension was heated to a gentle reflux for 20 minutes and then cooled to 0 ° C. Then n-butyllithium (40 ml, 2.5 M in hexanes) was added dropwise. The reaction was warmed to room temperature and then to 35 ° C for 30 minutes. The reaction was again cooled to 0 ° C and 3-picolyl chloride was added quickly. (The 3-picolyl chloride was obtained by neutralization of the hydrochloride with NaHCO 3 and extracted 3 times with hexane The hexane solution was dried over sodium sulfate and concentrated to provide the free base (caution: lachrymator) All hexane that it was not retired as the free base is something unstable in a concentrated form). The reaction was allowed to warm slowly to room temperature and stirred overnight. The reaction was quenched with H20, the aqueous layer was separated and washed twice with ether. The aqueous layer was then acidified to pH 3 with 6N HCl and again washed 2 times with ether. The aqueous phase was neutralized with sodium hydrogen carbonate and extracted 4 times with ethyl acetate. The organic extracts were combined, dried over magnesium sulfate, filtered and concentrated to give a sticky solid, which was triturated with ethyl acetate to provide 1.02 g of the desired product as a color-burned solid.

Example 2 2, 2-dimethyl-3- (3-pyridinyl) propionic acid N-oxide To a solution of 2,2-dimethyl-3- (3-pyridinyl) propionic acid (2.0 g, 11.17 mmol) in chloroform (48 ml) MCPBA (4.1 g, 13.07 mmol, 50% by weight) was added. The solution was stirred for 2.5 hours and then concentrated in vacuo. The residue was purified by means of flash column chromatography using 2-20% methanol in methylene chloride as eluent to give the 2,2-dimethyl-3- (3-pyridinyl) propionic acid N-oxide (1.88 g, 86%) as a white powder. NMR lH (200 MHz, DMSO) d 1.0 (s, 6H), 2.60 (s, 2H), 7.10 (m, 1H), 7.30 (m, 1H), 8.00 (s, 1H), 8. 05 (d, 1H), 12.5 (s, 1H). mp - 177-180 ° C Example 3 42-ester of rapamycin with 2,2-dimethyl-3- (3-pyridinyl) propionic acid N-oxide To a solution of 2,2-dimethyl-3- (3-pyridinyl) propionic acid N-oxide (1.02 g, 5.25 mmol) in tetrahydrofuran (36 ml) was added N, N-diisopropylethylamine (0.67 g, 5.25 mmol) followed by trichlorobenzoyl chloride (1.22 g, 5.02 mmol). The solution was stirred for 2 hours, and the solvent was removed by means of a stream of N2. Benzene (35 ml) was added followed by rapamycin (3.0 g, 3.28 mmol) and DMAP (0.64 g, 5.25 mmol). The reaction was stirred overnight and then quenched with sodium acid carbonate (sat). The aqueous solution was extracted with ethyl acetate, dried over sodium sulfate, concentrated and purified by flash column chromatography using 1-5% methanol in methylene chloride as eluent, followed by recrystallization from ethanol. water to provide 1.35 g, 38% of the title compound, mp = 183 ° C. IR (KBr) 1100 (m), 1160 (m), 1190 (m), 1275 (m), 1300 (m), 1325 (m), 1375 (m), 1450 (s), 1630 (s), 1725 (s), 2920 (s), 3420 (s); NMR XH (400 MHz, CDC13) d 0.80-1.95 (comp m, 21H), 0.91 (d, superimposed on comp m, J = 6.81 Hz, 3H), 0.95 (d, superimposed on comp m, J »6.37 Hz, 3H), 0.99 (d, superimposed on comp m, J = 6.37 Hz, 3H), 1.05 (d, superimposed on comp m, J - 6.59 Hz, 3H), 1.09 (d, superimposed on comp m, J = 6.81 Hz , 3H), 1.17 (s, superimposed on comp m, 3H), 1.24 (s, superimposed on comp m, 3H), 1.65 (s, superimposed on comp m, 3H), 1.76 (s, superimposed on comp m, 3H ), 2.11 (m, 4H), 2.32 (m, 3H), 2.59 (d, J = 6.37 Hz, 1H), 2.76 (m, 2H), 2.87 (m, 1H), 3.12-3.42 (comp m, 3H), 3.14 (s, superimposed on comp m, 3H), 3.33 (s, superimposed on comp m, 6H), • 3.57 (m, 1H), 3.68 (m, 1H), 3.75 (d, J - 5.71 Hz, 1H), 3.86 (m, 1H ), 4.19 (d, J - 5.93 Hz, 1H), 4.66 (m, 1H), 4.77 (s, 1H), 5.16 (m, 1H), 5.29 (m, 1H), 5.41 (m, 1H), 5.41 (d, J = 10.11 Hz, 1H), 5.53 (dd, J -8.79, 15.16 Hz, 1H), 5.97 (d, J - 10.55 Hz, 1H), 6.13 (dd, J = 9.89, 15.16 Hz, 1H) , 6.33 (m, 2H), 7.18 (s, 2H), 8.10 (s, 2H); 13C (100 MHz, CDC13) d 10.16, 13.22, 13.66, 15.87, 15.92, 16.24, 20.66, 21.49, 24.77, 25.58, 27.23, 29.63, 31.19, 32.81, 33.17, 33.77, 35.10, 35.70, 38.31, 38.94, 40.19, 40.49, 40.89, 41.51, 42.74, 43.33, 44.22, 46.60, 51.27, 55.89, 56.26, 57.09, 59.29, 67.18, 75.38, 76.36, 77.16, 80.97, 84.30, 84.40, 84.71, 86.34, 98.46, 125.25, 126.53, 128.13, 129.47, 130.17, 133.57, 135.67, 136.04, 137.34,. 140.09, 140.34, 140.71, 166.74, 169.25, 175.81, 192.63, 208.22, 215.29; high resolution mass spectrum (negative ion FAB) m / z 1090.7 [(M- «); calculated for C6? H9oN2O? 5: 1091.39].

Results obtained in standard pharmacological test procedures: LA5 IC5o: 2.9 nM Skin graft survival: i.p .: 11.17 ± 0.98 days; oral: 11 ± 0.89 days.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Having described the invention as above, property is claimed as contained in the following:

Claims

1. A composite of the structure characterized in that R and R1 are each, independently, , or hydrogen; R2 and R3 are each, independently, alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, or R2 and R3 can be taken together to form a cycloalkyl ring of 3 to 8 carbon atoms; R4 is a heterocyclic N-oxide radical of 5 to 12 carbon atoms, which may be optionally mono-, di-, or tri- substituted with a group selected from alkyl of 1 to 6 carbon atoms, arylalkyl of 7 to 10. carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7 carbon atoms, trifluoromethyl, trifluoromethoxy, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 carbon atoms carbon, -S03H, -P03H2, and -C02H; R5 is alkyl of 1 to 6 carbon atoms or arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms; R6 and R7 are taken together to form an N- (C 1-6 -alkyl) -N-oxide-heterocyclic ring of 5 to 8 atoms, which may optionally be mono- or tri-substituted. with a group selected from alkyl of 1 to 6 carbon atoms, aroyl of 3 to 11 carbon atoms, and perfluoroalkyl of 1 to 6 carbon atoms; k-0-1; m-0-1; n = 1-6; with the proviso that R and R1 are not both hydrogen.

2. The compound according to claim 1, characterized in that R1 is hydrogen.

3. The compound according to claim 2, characterized in that R is

4. The compound according to claim 3, characterized in that R4 is pyridinyl N-oxide, pyrazinyl N-oxide, triazinyl N-oxide, pyrimidinyl N-oxide, pyridazinyl N-oxide, imidazolyl N-oxide, N pyrazolyl oxide, quinolinyl N-oxide, or isoquinolinyl N-oxide.

5. The compound according to claim 4, characterized in that R 4 is pyridinyl N-oxide.

6. The compound according to claim 5, characterized in that k = 0.

7. The compound according to claim 1, characterized in that it is the 42-ester of rapamycin with the N-oxide of 2,2-dimethyl-3- (3-pyridinyl) propionic acid.

8. A method for the treatment of transplant rejection or graft versus host disease in a mammal in need thereof, characterized in that it comprises administering an anti-rejection effective amount of a compound of the structure where R and R1 are each, independently, , or hydrogen; R2 and R3 are each, independently, alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, or R2 and R3 can be taken together to form a cycloalkyl ring of 3 to 8 carbon atoms; R4 is a heterocyclic N-oxide radical of 5 to 12 carbon atoms, which may be optionally mono-, di-, or tri- substituted with a group selected from alkyl of 1 to 6 carbon atoms, arylalkyl of 7 to 10. carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7 carbon atoms, trifluoromethyl, trifluoromethoxy, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 carbon atoms carbon, -S03H, -P03H2, and -C02H; R5 is alkyl of 1 to 6 carbon atoms or arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms; R6 and R7 are taken together to form an N- (C 1-6 -alkyl) -N-oxide-heterocyclic ring of 5 to 8 atoms, which may optionally be mono- or tri-substituted. by a group selected from alkyl of 1 to 6 carbon atoms, aroyl of 3 to 11 carbon atoms, and perfluoroalkyl of 1 to 6 carbon atoms; k = 0 - 1. m = 0 - 1 n = 1 - 6, with the proviso that R and R1 are not both hydrogen.

9. A method for the treatment of rheumatoid arthritis in a mammal in need thereof, characterized in that the method comprises administering an effective anti-arthritis amount of a compound of the structure R and R1 are each, independently, O R - (CH ^ or hydrogen; R2 and R3 are each, independently, alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, or R2 and R3 can be taken together to form a cycloalkyl ring of 3 to 8 carbon atoms; R4 is a heterocyclic N-oxide radical of 5 to 12 carbon atoms, which may be optionally mono-, di-, or tri- substituted with a group selected from alkyl of 1 to 6 carbon atoms, arylalkyl of 7 to 10. carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7 carbon atoms, trifluoromethyl, trifluoromethoxy, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 carbon atoms carbon, -S03H, -P03H2, and -C02H; R5 is alkyl of 1 to 6 carbon atoms or arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms; R6 and R7 are taken together to form an N- (C 1-6 -alkyl) -N-oxide-heterocyclic ring of 5 to 8 atoms, which may optionally be mono- or tri-ring. substituted by a group selected from alkyl of 1 to 6 carbon atoms, aroyl of 3 to 11 carbon atoms, and perfluoroalkyl of 1 to 6 carbon atoms; k-0-1; m = 0-1; n-1 - 6; with the proviso that R and R1 are not both hydrogen.

10. A method for the treatment of pulmonary inflammation in a mammal in need thereof, characterized in that the method comprises administering to said mammal, an anti-inflammatory effective amount of a compound of the structure characterized in that R and R1 are each, independently, , or hydrogen; R and R are each, independently, alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, or R2 and R3 can be taken together to form a cycloalkyl ring of 3 to 8 carbon atoms; R4 is a heterocyclic N-oxide radical of 5 to 12 carbon atoms, which may be optionally mono-, di-, or tri- substituted with a group selected from alkyl of 1 to 6 carbon atoms, arylalkyl of 7 'to 10 carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7 carbon atoms, trifluoromethyl, trifluoromethoxy, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 atoms of carbon, -S03H, -P03H2, and -C02H; R5 is alkyl of 1 to 6 carbon atoms or arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms; R6 and R7 are taken together to form an N- (C 1-6 -alkyl) -heterocyclic N-oxide. from 5 to 8 atoms in the ring, saturated which may be optionally mono- or tri-substituted with a group selected from • alkyl of 1 to 6 carbon atoms, aroyl of 3 to 11 carbon atoms, and perfluoroalkyl of 1 to 6 carbon atoms; k = 0-1; m = 0-1; n = 1-6; with the proviso that R and R1 are not both hydrogen.

11. A method for the treatment of restenosis in a mammal in need thereof, characterized in that it comprises the administration to said mammal of an antiproliferative effective amount of a compound of the structure characterized in that R and R1 are each, independently, , or hydrogen; R2 and R3 are each, independently, alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, or R2 and R3 can be taken together to form a cycloalkyl ring of 3 to 8 carbon atoms; R4 is a heterocyclic N-oxide radical of 5 to 12 carbon atoms, which may be optionally mono-, di-, or tri- substituted with a group selected from alkyl of 1 to 6 carbon atoms, arylalkyl of 7 to 10. carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7 carbon atoms, trifluoromethyl, trifluoromethoxy, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 carbon atoms carbon, -S03H, -P03H2, and -C02H; R5 is alkyl of 1 to 6 carbon atoms or arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms; R6 and R7 are taken together to form an N- (C 1-6 -alkyl) -N-oxide-heterocyclic ring of 5 to 8 atoms, which may optionally be mono- or tri-substituted. with a group selected from alkyl of 1 to 6 carbon atoms, aroyl of 3 to 11 carbon atoms, and perfluoroalkyl of 1 to 6 carbon atoms; k-0-1, m-0-1 n-1-6, with the proviso that R and R1 are not both hydrogen.

12. A method for the treatment of a fungal infection in a mammal in need thereof, characterized in that the method comprises administering to said mammal an effective antimycotic amount of a compound of the structure characterized in that R and R are each, independently, , or hydrogen; R2 and R3 are each, independently, alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, or R2 and R3 can be taken together to form a cycloalkyl ring of 3 to 8 carbon atoms; R4 is a heterocyclic N-oxide radical of 5 to 12 carbon atoms, which may be optionally mono-, di-, or tri- substituted with a group selected from alkyl of 1 to 6 carbon atoms, arylalkyl of 7 to 10. carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7 carbon atoms, trifluoromethyl, trifluoromethoxy, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 atoms of carbon, -S03H, -P03H2, and -C02H; R3 is alkyl of 1 to 6 carbon atoms or arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms; R6 and R7 are taken together to form an N- (C 1-6 -alkyl) -N-oxide-heterocyclic ring of 5 to 8 atoms, which may optionally be mono- or tri-substituted. with a group selected from alkyl of 1 to 6 carbon atoms, aroyl of 3 to 11 carbon atoms, and perfluoroalkyl of 1 to 6 carbon atoms; k = O-1; m = O-1; n = 1-6; with the proviso that R and R1 are not both hydrogen.

13. A pharmaceutical composition, characterized in that it comprises a compound of the structure characterized in that R and R1 are each, independently, , or hydrogen; R2 and R3 are each, independently, alkyl of 1 to 6 carbon atoms, arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms, or R2 and R3 can be taken together to form a cycloalkyl ring of 3 to 8 carbon atoms; R4 is a heterocyclic N-oxide radical of 5 to 12 carbon atoms, which may be optionally mono-, di-, or tri- substituted with a group selected from alkyl of 1 to 6 carbon atoms, arylalkyl of 7 to 10. carbon atoms, alkoxy of 1 to 6 carbon atoms, cyano, halo, hydroxyl, nitro, carbalkoxy of 2 to 7 carbon atoms, trifluoromethyl, trifluoromethoxy, hydroxyalkyl of 1 to 6 carbon atoms, alkoxyalkyl of 2 to 12 atoms of carbon, -S03H, -P03H2, and -C02H; R5 is alkyl of 1 to 6 carbon atoms or arylalkyl in which the alkyl portion contains from 1 to 6 carbon atoms; R6 and R7 are taken together to form an N- (C 1-6 -alkyl) -N-oxide-heterocyclic ring of 5 to 8 atoms, which may optionally be mono- or di-substituted. with a group selected from alkyl of 1 to 6 carbon atoms, aroyl of 3 to 11 carbon atoms, and perflubroalkyl of 1 to 6 carbon atoms; J - 0 - 1. m - 0 - 1 n - 1 - 6, with the proviso that R and R1 are not both hydrogen.

14. A process for the preparation of the hindered N-oxide-esters of rapamycin, including those of the formula I according to claim 1, characterized the process because it comprises: a) the acylation of rapamycin or a functional derivative thereof with an acylating agent or b) the sequential acylation of rapamycin or a functional derivative thereof with two acylating agents said acylating agents are selected from the acids of the formula or < IIa > (Ilb) wherein n, k, m and R2-R7 are as defined above, or a reactive derivative thereof, if desired protecting the 42 position of rapamycin with an appropriate protecting group and eliminating it as required. SUMMARY OF THE INVENTION A compound of structure (I) is described, wherein R and R1 are each, independently, a), b), or hydrogen; R2 and R3 are each, independently, alkyl, arylalkyl, or R2 and R3 can be taken together to form a cycloalkyl ring; R 4 is a heterocyclic N-oxide radical, which may be optionally substituted; R5 is alkyl or arialkyl; R6 and R7 are taken together to form a saturated N-alkyl-heterocyclic N-oxide, which may be optionally substituted; k = »0-1, -0-1; n-1 - 6; with the proviso that R and R1 are not both hydrogen, which is useful as an immunosuppressive, anti-inflammatory, antimycotic, antiproliferative and oral antitumor agent. (to) (b) «Aßn» ^ na »npn» »«?