JP2001503061A - Synergistic composition comprising rapamycin and calcitriol - Google Patents

Abstract

The present invention is useful for inducing immunosuppression, including transplant rejection, graft-versus-host disease, autoimmune disease, inflammatory disease, adult T-cell leukemia / lymphoma, tumor, fungal infection and useful rapamycin and 1,25-dihydroxy the treatment of hyperproliferative vascular disorders - provides a combination of cholecalciferol _{(1,25 (OH) 2 D 3} ).

Description

DETAILED DESCRIPTION OF THE INVENTION         Synergistic composition comprising rapamycin and calcitriol   Background of the Invention   In recent years, the use of immunosuppressants can prevent or treat many diseases or disorders. Was recognized. In particular, the use of immunosuppressants can result in organ transplantation It has become a successful treatment for many serious conditions that can be fatal. Or What used to be an experimental method used only in emergency life-threatening situations, It has been used early in the treatment of patients with severe chronic illness. Currently, kidney, heart, Allogeneic transplantation between lung, liver, bone marrow, pancreas (islet cells), cornea, small intestine and skin It is done conventionally. In addition, xenografts are currently performed using porcine heart valves .   Despite advances in organ allotransplantation, transplant rejection can lead to transplant failure Powerful factors remain. Overall, transplant rejection is a complex process involving the immune system is there. This process is briefly outlined below. Rejection moves from allograft Allograft blood-derived antigen presenting cells (dendrites expressing class II MHC molecules) Cells and monocytes). Antigen recognition by antigen presenting cells Production of IL-1 and CD-1⁺Causes activation of T cells, thereby Initiate an immune response leading to eventual transplant rejection. Activated CD4⁺T cells are C D8⁺Produces IL-2, a growth factor required for activation of both T cells and B cells Live. Is clonal expansion and maturation of alloantigen-reactive cells a host lymphoid tissue? Effector T cells that migrate from and infiltrate the transplanted tissue (cytotoxic CD8⁺T cells And CD4⁺T cells). The infiltration of T lymphocytes into the vascular endothelium Attachment, migration through the vessel wall, migration within the graft, selection of activated cells within the graft Includes alternative retention and local proliferation of cells. Antigen presenting transplanted cells are cytotoxic CD8⁺ Directly destroyed by T cells. In addition, CD4⁺T cells also contribute to graft destruction Such as interferon-γ (IFN-γ), IL-4 and IL-5 Produces other lymphokines. IFN-γ is used in transplanted tissues for HLA-A, Effector mechanism causing increased expression of B and -DR More vulnerable to attacks. IFN-γ also activates macrophages. To initiate a delayed sensitive reaction that causes non-specific damage to the graft. I L-4 and IL-5 show antibody production by plasma cells leading to antibody-mediated damage of the graft Related to triggering. [Hutchinson, I., Transplantation 3: 722 (1991); Garovoy , M .; R., Basic and Clinical Immunology, ed. Stites, 7thed., 747 (1991)]   Currently, allograft rejection includes prednisone, methylprednisolone, azathiop Phosphorus, cyclophosphamide, cyclosporin, anti-lymphocyte globulin, monochrome It is controlled using an internal antibody and an agent that suppresses the immune response, such as irradiation. Shi Crossporin A is the most efficacious and most frequent chemotherapeutic agent currently used. Although commonly used, it has unsatisfactory side effects of nephrotoxicity in humans, May cause kidney damage.   Recently, rapamycin, Streptomyces hygroscopicus (Streptomyc es hygroscopicus) produced by macrocyclic triene antibiotics [rice No. 3,929,992] responds to an albumin allergic attack. To prevent the formation of humoral (IgE-like) antibodies [Martel, R., Can. J. Physio l. Pharm. 55:48 (1977)], which inhibits murine T cell activation [Staruch, M., FASEB 3: 3411 (1989)], extending the survival time of organ transplantation in tissue-incompatible rodents [Morris, R., Med. Sci. Res. 17: 877 (1989)], transplant rejection in mammals [US Patent No. 5,100,999]. Rapamycin Is also known as pulmonary inflammation [US Pat. No. 5,080,899], systemic lupus erythematosus Des [US Pat. No. 5,078,899], immunoinflammatory skin disorders such as dry Iris [US Pat. No. 5,286,730], Immune inflammatory bowel disorder [US Pat. 86,731], eye inflammation [US Pat. No. 5,387,589], hyperproliferative blood Vascular disorders, such as restenosis [U.S. Patent Nos. 5,512,781 and 5,288, 711], carcinoma [US Pat. Nos. 5,206,018 and 4,885,171. No.] and treatment of cardiac inflammatory disease [US Pat. No. 5,496,832]; Prevention of insulin-dependent diabetes mellitus [US Patent No. 5,321,009] Has been shown to be useful. In addition, rapamycin is It has been shown to be useful in the treatment of human T-cell leukemia / lymphoma [European Patent Application No. 525, 960 A1].   Vitamin D, 1,25-dihydroxycholecalciferol (1,25 (OH )_TwoD_Three), Its active form in bone metabolism and calcium / phosphate homeostasis In addition to its typical role, it also shows non-calcemic effects [Walters, M .; R. , Endocr. Rev. 13: 719 (1992)]. For example, 1,25 (OH)_TwoD_ThreeProbably, Immunity acted through specific receptors present on both monocytes and activated T lymphocytes It has a strong influence on the epidemic system [Rigby, W.F.C., Immunol. Today 9:54 (198 8)].   1,25 (OH)_TwoD_ThreeIs immune at macrophage and T helper levels Reduce cell interactions [Rigby, W.F.C., Blood 76: 189 (1990)]. Regulates paracrine function [Manolagas, S.C., Semin. Nephrol. 14: 129 (1994) It seems. IL-2 [Muller, K., Immunol. Lett. 35: 177 (1993)], IF N-γ [Reichel, H., Proc. Natl. Acad. Sci. USA 84: 3385 (1987)] or IL -12 [Lemire, J .; M., in Vitamin D, a pluripotent steroid hormone: struc tural studies, molecular endocrinology, and clinical applications, A. W . Norman ed, de Druyter, W.S. Berlin, p. 534 (194)] -1,25 (OH) of leukin secretion_TwoD_ThreeDirect inhibition by And recruitment of other immune effectors. These in vitro data are , And further supported by animal studies. Actually, 1,25 (OH)_TwoD_Threeand Some of the analogs prevent the severity of some autoimmune diseases or Can be reduced and skin or heart allograft survival can be prolonged [ Lemire, J.M., J. Cell Biochem. 49: 326 (1992)]. However, therapeutically effective Dose treatment with vitamin D-related drugs is accompanied by toxic hypercalcemic effects [Binderup, L., Biochem. Pharmacol. 43: 1885 (1992)].   Description of the invention   The present invention is useful for inducing immunosuppression, transplant rejection, graft-versus-host disease, autoimmune Epidemics, inflammatory diseases, adult T-cell leukemia / lymphoma, carcinomas, fungal infections and hyperplasia And 1,25-dihydroxycholeca useful for the treatment of reproductive vascular disorders Luciferol (1,25 (OH)_TwoD_Three) To provide a combination.   When administered to treat or inhibit the above conditions, rapamycin and 1, 25 (OH)_TwoD_ThreeIs orally, parenterally, It can be administered intranasally, intrabronchially, transdermally, topically, vaginally or rectally.   The present invention also relates to rapamycin and 1,25 (OH)_TwoD_ThreeAnd a pharmaceutical carrier A pharmaceutical composition comprising:   Rapamycin and 1,25 (OH)_TwoD_ThreeCombined immunosuppressants The ability to use in vitro and in vivo standard pharmacology It has been established. In vitro procedures include rapamycin and 1,25 (OH)_TwoD_Three Were evaluated for their ability to inhibit PHA-stimulated PMBC proliferation. Multiple hard In vivo test methods that mimic keratosis and autoimmune diseases Mycin and 1,25 (OH)_TwoD_ThreeCombination of experimental allergic cerebrospinal cord The ability to inhibit the development of flame was measured. The methods used and the results obtained are described below. Will be described briefly.   Using the PHA-stimulated PMBC proliferation standard pharmacology test method, rapamycin and 1,25 (OH)_TwoD_ThreeThe immunosuppressive activity of the combination was evaluated. Put simply, Human PBMCs were healthy by Ficoll-Hypaque density gradient centrifugation. Isolated from freshly collected heparin-inhibited venous blood of healthy volunteers. Cells at a density of 10⁶/ Ml 10% heat-inactivated fetal bovine serum, L-glutamine ( 2 mM), penicillin (100 U / ml), streptomycin (100 μg / ml) and mycostatin (100 U / ml) in RPMI medium. Suspended, 5% CO_TwoPhytohemagglutinin (PHA, 0.1 μg / ml), 1,25 (OH)_TwoD_Three(10^-9Or 3.125 × 10^-6 M), rapamycin (0.32-1000 ng / ml, ie 3.50 1x10^-TenTo 1.094x10^-6M), vehicle control or 1,25 (OH )_TwoD_ThreeAnd a combination of serial dilutions of rapamycin. 0.2ml -Bottom multi-well plates containing the final cell suspension in each well (NUNC, Denm ark). Rapamycin and 1,25 (OH)_TwoD_ThreeIs always with PHA Added to PBMC culture 20-30 minutes before addition. Each individual concentration or pair The combination as well as the control were tested four times. After 48 hours of incubation, each well Is 0.5 pCi [^ThreeH] -Thymidine. Plate for another 18 hours , Reincubated and then harvested using a microwell harvester. The radioactivity was measured using a β-scintillation counter and the percent growth was counted. Calculated.   The results obtained with the standard pharmacology test method were compared to the specified PHA-stimulated PMBC proliferation. The following table shows the dosages required to produce a percent inhibition of                       Percent inhibition of PMBC proliferation   These results indicate that rapamycin, 1,25 (OH)_TwoD_ThreeAnd rapamycin And 1,25 (OH)_TwoD_ThreeAll combinations of PHA-stimulated PMBC proliferation Indicates inhibition. Rapamycin alone has an IC of 180 ng / ml₅₀Has , 1,25 (OH)_TwoD_Three2.2x10 alone^-FiveM IC₅₀But with Rapama Isin and 1,25 (OH)_TwoD_Three1.7 ng / m 1 rapamycin and 5.2 x 10^-9M1, 25 (OH)_TwoD_ThreeIC at dose of₅₀ Reached. The combinations were rapamycin and 1,25 (OH), respectively._TwoD_ThreeAlone More than 100 times less than 4000 times treatment gave 50% inhibition . These results indicate that rapamycin and 1,25 (OH)_TwoD_ThreeCombination free It indicates that it acted synergistically as an epidemic inhibitor.   Experimental allergic encephalomyelitis (EAE) using standard pharmacological test methods Isin and 1,25 (OH)_TwoD_ThreeTo further evaluate the immunosuppressive activity of the combination Was. The test method also evaluated the activity of the combination against autoimmune diseases. Briefly, mouse spinal cord isolated from NMRI mice was lyophilized into powder. And stored at + 4 ° C. Mouse spinal cord and mycobacterium in PBS (55 mg / ml) Complete adjuvant of Freund's Freund of Terium tuberculosis strain H37-Ra (CFA, 4 mg / Ml) of a 50 μl emulsion of the equal mixture in Injected. The day of induction was considered day 0. On days 0 and +2, 2 in a 50 μl PBS volume 00 ng of pertussis toxin was given intravenously on the day of induction and repeated 2 days later.   Control mice (group CTR = 42 mice) were induced and 100 μl of Rapamy Symvehicle (0.2% carboxymethylcellulose in phosphate buffered saline) ) Every day, 1,25 (OH)_TwoD_ThreeInject vehicle (100 μl peanut oil) every 2 days Fired. The next three groups start on day -3 (three days prior to challenge) and start every other day with 1 1 μg / kg (Group D1 = 21 mice), 21 μg / kg (Group D2 = 23 mice) and 5 μg / kg (group D5 = 20 mice) at 1,25 ( OH)_TwoD_ThreeOnly received. The group treated with rapamycin alone received 0,0 daily. 1 mg / kg (group RP 0.1 = 25 mice), 0.3 mg / kg (group RP RP 0.3 = 36 mice), 0.6 mg / kg (group RP 0.6 = 32 mice) Mouse) and 1 mg / kg (group RP1 = 28 mice) or every 2 days 0.6 mg / kg (Group RP 0.6 '= 35 mice) Was. The last two groups are 2 pg / kg of 1,25 (OH) every 2 days_TwoD_Threeand Although treated with the same dose of rapamycin combination, the daily dose of 0.3 mg / kg ( Group MIX-1, 25 mice) or 0.6 mg / kg for 2 days (Group MIX-2, 23 mice). Of the last group Rapamycin administration started rapamycin in -3 days and was 1,25 (OH)_TwoD_Threeof Alternating with dosing. Three to seven urine samples were collected from each mouse and urinary calcium And collagen crosslink content (deoxypyridinoline, DPD). All mice were killed on day 20 or earlier if severely paralyzed. blood Fluid was collected by cardiac puncture under ether paralysis. Calcium and osteo Serum was stored at -20 <0> C for further measurement of calcin. Duodenum, 1c The pylorus is isolated and rinsed, and calbindin D-9K content Were stored at -20 ° C until assayed. Remove the brain and spinal cord and For immobilization in Bouin's solution.   On day 9, the mice were evaluated for clinical signs of paralysis. 0 to 4 EAE signatures were recorded daily on a scale of. Score is two independent observers Was blindly evaluated by To be positive for clinical EAE Mice must score 0.5 or higher for two consecutive days. No. In the case of a mouse that has died after giving the disease sign in advance, or In the case of moribund mice killed before 20 days, a score of 4 from their death date It was considered until the follow-up was over. When comparing disease development in different groups, Disease free survival indicating the number of disease free mice in a group at the day of sacrifice, And the following parameters of disease severity: 1 group for 11-20 days of follow-up Cumulative score defined as the sum of daily average scores for Integrated disease score calculated by daily score average of all individuals for the next 10 days And the average of the highest disease scores achieved by all mice in one group The average maximum disease score was used.   The result of the standard pharmacological test method is 1,25 (OH)_TwoD_ThreeAnd rapamycin alone Both treatments in Germany showed significantly reduced disease severity (ANOVA P <0.0001 by test). All but one control mouse showed signs of paralysis Indicated. Disease onset was observed 9-15 days after induction (mean 12.1 ± 0.3 days ). The disease is generally more severe until the peak score is reached on days 1 to 4. Mice then either died or recovered slowly. A similar rating is the best Also a small dose of 1,25 (OH)_TwoD_ThreeAnd rapamycin treated groups (Group D1 and RPO.1). Higher drug dose Revealed a more substantial protective effect. 1,25 (2 μg / kg every two days OH)_TwoD_Three8 of 23 mice were followed up in the group treated with No disease at the end of the day (30%, group D2) every 5 days at 5 μg / kg The mice treated with 防止 prevent the appearance of disease in 65% of the treated mice (25 13 of 13 mice, group D5), disease in mice that developed disease Onset was delayed (p <0.01). Rapamycin (glue) at 1 mg / kg / day RP1) gave almost absolute protection (only 2 out of 28 animals) Developed milder illness), and lower doses were only partially protected. Was. For example, rapamycin at 0.3 mg / kg / day is found in 45% of animals. To prevent disease development (Group RP0.3), but given the same amount of drug every two days Then (0.6 mg / kg every 2 days, group RP 0.06 '), somehow Less protection (31% disease-free animals at the end of the observation period).   Subtherapeutic dose of 1,25 (OH)_TwoD_ThreeAnd rapamycin combination (Groups MIX-1 and MIX-2) were obtained in group RP1 Provided almost absolute protection comparable to protection. Achieved with combination therapy The protection was much more effective than obtained with each drug separately.   These results are summarized in the table below.                           Clinical evaluation of EAE  The severity of brain and spinal cord inflammation is generally determined by the clinical Corresponding to the core. Alleviation of severe monocyte infiltration was reduced in 7 of 8 control mice Observed, mild inflammation was still present in the eighth animal. Mild central nervous system The number of mice without inflammation or central nervous system inflammation were group D2, RP0.3 and RP 0.6 'was 4, 5 and 3 out of 8 respectively. Both sets Of the 8 mice in the combined treatment group and group RP1 Seven had no inflammation and only one showed monocyte infiltration. The average histological score is Significantly lower than group D2 scores in both combination groups . The score of group MIX-2 is also higher than the score of group RP0.6 '. Although significantly lower, the difference between the scores of groups MIX-1 and RP0.3 was The significance limit was reached (p = 0.06).   These results are summarized in the table below.                       Mean histological score and inflammation* Number of animals with CNS inflammation   The results obtained in the standard pharmacology test method were rapamycin and 1,25 (OH)_TwoD_ThreeIs a combination of immunosuppressants, experimental allergic encephalomyelitis, Demonstrates synergy in inhibiting the formation of standard models of autoimmune disease did.   Based on the results obtained in standard pharmacological test methods, rapamycin and 1 , 25 (OH)_TwoD_ThreeCombination for the treatment of diseases or disorders involving the immune system or Useful for prevention. These include, but are not limited to, kidney, heart, liver, lung, bone marrow , Pancreas (islet cells), cornea, small intestine and skin allograft, and heart valve xenograft Treatment or inhibition of transplant rejection, such as chemotherapy; systemic lupus erythematosus, rheumatoid arthritis Gusset, diabetes, myasthenia gravis, aplastic anemia, pure erythrocyte anemia, idiopathic platelets Hypotension, polychondritis, palmar dermatosis, Wegener's granulomatosis, chronic active hepatitis, bile liver cirrhosis Strange Sarcoidosis, nephrotic syndrome, multiple sclerosis, Steven Soo John Song syndrome, psoriasis, dermatitis, eczema, seborrhea, idiopathic sprue, Crohn's disease, inflammation Autoimmune diseases such as genital bowel disease, Graves' eye disease, and interstitial pulmonary fibrosis; Inflammation diseases; eye inflammation such as uveitis; asthma, chronic obstructive pulmonary disease, emphysema, sudden Inflammation of the lungs, such as dyspnea syndrome and bronchitis; and, in particular, the intima Restenosis following biological or mechanically mediated vascular injury leading to smooth muscle cell proliferation Includes treatment or inhibition of hyperproliferative vascular disorders such as stenosis, vascular occlusion. Lapamai Syn and 1,25 (OH)_TwoD_ThreeAre also fungal infections and adult T Useful as an antitumor agent for the treatment of cell leukemia / lymphoma.   Rapamycin and 1,25 (OH)_TwoD_ThreeCombination achieves the desired effect Diseases or disorders involving the immune system to further reduce the dosage required to Other drugs useful in the treatment or prevention of azathioprine, corticostero , Such as prednisone and methylprednisolone, cyclophosphamide , Cyclosporin A, FK-506, OKT-3, and ATG in combination It is expected that it may be used.   Rapamycin and 1,25 (OH)_TwoD_ThreeInduces immunosuppression using a combination of If fired, the mammal, in need thereof, in a straight or pharmaceutical carrier Can be prescribed with Pharmaceutical carriers can be solid or liquid.   Solid carriers include flavoring agents, lubricants, solubilizing agents, suspending agents, fillers, lubricants, Shrinkage aids, binders or tablets-including one or more substances that can also act as disintegrants Can also be an encapsulation material. In powders, the carrier is fine powder It is a finely divided solid mixed with the active ingredient. In tablets, the active ingredient is present in suitable proportions It is mixed with a carrier having the required compression properties and formed into the desired shape and size. Scattered The agents and tablets preferably contain up to 99% of the active ingredient. Suitable solid carrier For example, calcium phosphate, magnesium stearate, talc, sugar, la Kutose, dextrin, starch, gelatin, cellulose, methylcellulose , Sodium carboxymethylcellulose, polyvinylpyrrolidine, low melting wax And ion-exchange resins.   Liquid carriers include solutions, suspensions, emulsions, syrups, elixirs and pressurized Used to prepare the composition. The active ingredient is a pharmaceutically acceptable liquid carrier, such as water , An organic solvent, a mixture of both pharmaceutically acceptable oils or fats, or either Can be dissolved or suspended in water. Liquid carriers may contain other suitable pharmaceutical excipients, for example, solubilized Agents, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickeners, coloring It may contain additives, viscosity modifiers, stabilizers or penetration modifiers. Oral and parenteral A suitable example of a liquid carrier for administration is water (excipient such as a cellulose derivative, Preferably, it partially contains a sodium carboxymethylcellulose solution. Alcohols (monohydric and polyhydric alcohols such as glycols) And their derivatives, and oils (eg, fractionated coconut oil and Peanut oil). For parenteral administration, the carrier may also be ethyl oleate or the like. And oily esters such as isopropyl myristate. Sterile liquid carrier Is useful in sterile liquid form compositions for parenteral administration. For pressurized compositions The liquid carrier can be a halogenated hydrocarbon or other pharmaceutically acceptable propellant .   Liquid pharmaceutical compositions that are sterile solutions or suspensions can be, for example, intramuscular, intraperitoneal or It can be used by subcutaneous injection. Sterile solutions may also be administered by intravenous injection. You can also. The compounds may also be in the form of either liquid or solid compositions. It can be administered orally.   Rapamycin and 1,25 (OH)_TwoD_ThreeCombination of traditional suppository forms May be administered rectally. Intranasal or intrabronchial inhalation (inhalation or insuff lation), the compounds of the invention are administered in aqueous or partially aqueous solution. And then available in aerosol form. Rapamycin and 1, 25 (OH)_TwoD_ThreeIs also a combination for the active compound and the active compound Inert, non-toxic to the skin, for systemic absorption through the skin and into the bloodstream By using a transdermal patch containing a carrier that enables the delivery of the drug, It may be administered transdermally. Carriers include creams and ointments, pastes, gels, and It can take many forms, such as an occlusive device. Creams and ointments may be used as viscous liquids or May be an oil-in-water or water-in-oil semi-solid emulsion. Contains absorbent powder dispersed in petroleum containing active ingredients or hydrophilic petroleum A paste consisting of the same may also be suitable. To release the active ingredient into the bloodstream, Storage tank containing active ingredient with or without carrier or containing active ingredient A variety of occlusive devices may be used, such as semipermeable membranes that cover the matrices to be formed. Other occlusive devices are known in the literature.   Rapamycin and 1,25 (OH)_TwoD_ThreeAre pharmaceutically acceptable 0.1-5%, preferably 2% activation by formulation with vehicle Topically administered as a solution, cream, or lotion containing the compound. No.   Dosage requirements will depend on the particular composition used, the route of administration, the severity of the existing condition, and the And with the individual being treated. Results obtained in standard pharmacological test methods And rapamycin and 1,25 (OH)_TwoD_ThreeThe combination is planned Daily intravenous doses were 0.001-25 mg / kg rapamycin and 0.1 mg / kg. 001-50 μg 1,25 (OH)_TwoD_ThreeWill. Rapamycin and 1,2 5 (OH)_TwoD_ThreeThe planned daily oral dose of the combination of mg / kg rapamycin and 0.01-100 μg 1,25 (OH)_TwoD_ThreeIn Would.   Generally, treatment will be initiated with smaller dosages which are less than the optimum dose of the compound. U. Thereafter, the dosage is increased until the optimum effect under the circumstances is reached; Precise dosages for oral, nasal, intrabronchial, transdermal or rectal administration may vary Will be determined by the administering physician based on the experience of the subject being treated. General Rapamycin and 1,25 (OH)_TwoD_ThreeThe combination of Provide effective results without causing any harmful or psychological side effects Will be most desirably administered.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 29/00 A61P 29/00 31/10 31/10 35/00 35/00 35/02 35/02 37 / 06 37/06 (81) Designated countries EP (AT, BE, CH, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA ( BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AP (GH, KE, LS, MW, SD, SZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, I, GB, GE, GH, HU, IL, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, UZ, VN, YU, ZW (72) Invention Matthew, Chantal Belgium, Be-1501 Weisingen, Klabos 38

Claims (1)

[Claims]   1. The mammal in need may include rapamycin and 1,25-dihydroxy Oral, parenteral, intranasal, intrabronchial combination of calciferol Induces immunosuppression in mammals consisting of transdermal or rectal administration How to emit.   2. The mammal in need may include rapamycin and 1,25-dihydroxy Oral, parenteral, intranasal, intrabronchial combination of calciferol Organ or set in a mammal, comprising administering transdermally or rectally A method of inhibiting or treating tissue transplant rejection.   3. The transplanted organ or tissue is kidney, heart, liver, lung, bone marrow, pancreas (islet cells), 3. The method of claim 2, wherein the method is selected from the group consisting of cornea, small intestine, skin and heart valve.   4. The mammal in need may include rapamycin and 1,25-dihydroxy Oral, parenteral, intranasal, intrabronchial combination of calciferol Autoimmune disease in mammals, comprising administering transdermally or rectally A method of treating or inhibiting the disease.   5. Combination of rapamycin and 1,25-dihydroxycholecalciferol The combination is administered to the mammal orally, parenterally, intranasally, intrabronchially, or transdermally. Treats or inhibits an inflammatory disease or disorder consisting of oral or rectal administration Method.   6. Combination of rapamycin and 1,25-dihydroxycholecalciferol The combination is administered to the mammal orally, parenterally, intranasally, intrabronchially, or transdermally. For treating or inhibiting hyperproliferative vascular disorders, comprising administering to a patient or the rectum .   7. Combination of rapamycin and 1,25-dihydroxycholecalciferol The combination is administered to the mammal orally, parenterally, intranasally, intrabronchially, or transdermally. Or a method for treating or inhibiting a tumor, comprising administering to the rectum.   8. Rapamycin, 1,25-dihydroxycholecalciferol and pharmaceutical A pharmaceutical composition comprising a carrier.