EP0549727A1 - Carboxylic acid esters of rapamycin - Google Patents

Carboxylic acid esters of rapamycin

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Publication number
EP0549727A1
EP0549727A1 EP19910919248 EP91919248A EP0549727A1 EP 0549727 A1 EP0549727 A1 EP 0549727A1 EP 19910919248 EP19910919248 EP 19910919248 EP 91919248 A EP91919248 A EP 91919248A EP 0549727 A1 EP0549727 A1 EP 0549727A1
Authority
EP
European Patent Office
Prior art keywords
carbon atoms
alkyl
rapamycin
compound
acceptable salt
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP19910919248
Other languages
German (de)
French (fr)
Inventor
Craig Eugene Caufield
Amedeo Arturo Failli
Robert John Steffan
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wyeth LLC
Original Assignee
American Home Products Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US07/657,294 external-priority patent/US5130307A/en
Application filed by American Home Products Corp filed Critical American Home Products Corp
Publication of EP0549727A1 publication Critical patent/EP0549727A1/en
Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
    • C07D498/18—Bridged systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/01—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing oxygen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06026—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to novel esters of rapamycin and a method for using them in the treatment of transplantation rejection, host vs. graft disease, autoimmune diseases, diseases of inflammation, and fungal infections.
  • Rapamycin is a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus. which was found to have antifungal activity, particularly against Candida albicans. both in vitro and in vivo [C. Vezina et al., J. Antibiot. 28, 721 (1975); S.N. Seghal et al., J. Antibiot. 28, 727 (1975); H. A. Baker et al., J. Antibiot. 31, 539 (1978); U.S. Patent 3,929,992; and U.S. Patent 3,993,749].
  • Rapamycin alone (U.S. Patent 4,885,171) or in combination with picibanil (U.S. Patent 4,401,653) has been shown to have antitumor activity.
  • R. Mattel 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.
  • rapamycin has been shown to be effective in inhibiting transplant rejection (U.S. Patent Application Ser. No. 362,544 filed June 6, 1989).
  • Cyclosporin A and FK-506, other macrocyclic molecules also have been shown to be effective as immunosuppressive agents, therefore useful in preventing transplant rejection [FASEB 3, 3411 (1989); FASEB 3, 5256 (1989); and R. Y. Calne et al., Lancet 1183 (1978)].
  • Mono- and diacylated derivatives of rapamycin (esterified at the 28 and 43 positions) have been shown to be useful as antifungal agents (U.S.
  • Patent 4,316,885 and used to make water soluble prodrugs of rapamycin (U.S. Patent 4,650,803). Recently, the numbering convention for rapamycin has been changed; therefore according to Chemical Abstracts nomenclature, the esters described above would be at the 31- and 42- positions. DESCRIPTION OF THE INVENTION
  • This invention provides derivatives of rapamycin which are useful as immunosuppressive, anti-inflammatory, and antifungal agents having the structure
  • R 1 , R 2 > and R 3 are each, independently, hydrogen, or R 4 ;
  • R 4 is -[C(CH 2 ) ni CH(CH 2 ) n N] p C0 2 R 7 , -C- (CH 2 ) t X(CH 2 ) u CO 2 R 11 , or
  • R 5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms,
  • R 6 and R 9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms;
  • R 7 , R 8 , and R 10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri- substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
  • R 1 ! and R 12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
  • R 13 and R 14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms; Y is CH or N; m is 0 - 4; n is 0 - 4; p is 1 - 2 q is 0 - 4 r is 0 - 4; t is O - 4; u is 0 - 4;
  • R5 R 6 subunits when p 2; or a pharmaceutically acceptable salt thereof, with the proviso that R 1 , R 2 * and R 3 are not all hydrogen, further provided that R 1 , R 2 - and R 3 are not all
  • R 5 R 6 both 0 when X is O or S.
  • R 4 is — [C(CH 2 ) m CH(CH 2 )-.N] p CO 2 R 7
  • Preferred compounds also include those
  • the pharmaceutically acceptable salts may be formed from inorganic cations such as sodium, potassium, and the like; mono-, di-, and trialkyl amines of 1-6 carbon atoms, per alkyl group and mono-, di-, and trihydroxyalkyl amines of 1-6 carbon atoms per alkyl group; and organic acids such as acetic, lactic, citric, tartaric, succinic, maleic, malonic, gluconic, and the like.
  • Preferred basic salts are formed from sodium cations and tris(hydroxymethyl)methyl-.mine.
  • the compounds of this invention can be prepared by acylating rapamycin with an acylating agent having the general structures
  • the compounds of this invention also can be prepared using an anhydride or a mixed anhydride of the above described carboxylic acid as the acylating species.
  • the acylating species can be an acid halide, where Z can be Cl, Br, or I.
  • the acylating groups used to prepare the compounds of this invention are commercially available or can be prepared by methods that are disclosed in the literature.
  • acyl derivatives having two or three different R 4 groups sequential acylation may be performed using appropriate acylating agents as defined above, if necessary isolating the desired product by appropriate purification techniques.
  • the 42-position is acylated first and such a monoacylated product may be isolated prior to the second acylation and so forth.
  • Appropriate protecting groups may be used to block any position where acylation is not required.
  • Immunosuppressive activity was evaluated in an in vitro standard pharmacological test procedure to measure lymphocyte proliferation (LAF) and in two in vivo standard pharmacological test procedures.
  • the first in vivo procedure was a popliteal lymph node (PLN) test procedure which measured the effect of compounds of this invention on a mixed lymphocyte reaction and the second in vivo procedure evaluated the survival time of a pinch skin graft.
  • the comitogen-induced thymocyte proliferation procedure (LAF) was used as an in vitro measure of the immunosuppressive effects of representative compounds. Briefly, cells from the thymus of normal B ALB/c mice are cultured for 72 hours with PHA and IL-1 and pulsed with tritiated thymidine during the last six hours.
  • Cells are cultured with and without various concentrations of rapamycin, cyclosporin A, or test compound. Cells are harvested and incorporated; radioactivity is determined. Inhibition of lymphoproliferation is assessed in percent change in counts per minute from non-drug treated controls. The results are expressed by the following ratio, or as the percent inhibition of lymphoproliferation of 1 ⁇ M. 3 H-control thvmus cells - H 3 -rapamvcin-treated thvmus cells
  • a mixed lymphocyte reaction occurs when lymphoid cells from genetically distinct animals are combined in tissue culture. Each stimulates the other to undergo blast transformation which results in increased DNA synthesis that can be quantified by the incorporation of tritiated thymidine. Since stimulating a MLR is a function of disparity at Major Histocompatibility antigens, an in vivo popliteal lymph node (PLN) test procedure closely correlates to host vs. graft disease. Briefly, irradiated spleen cells from B ALB/c donors are injected into the right hind foot pad of recipient C3H mice. The drug is given daily, p.o. from Day 0 to Day 4.
  • tritiated thymidine is given i.p., b.i.d.
  • the hind popliteal lymph nodes are removed and dissolved, and radioactivity counted.
  • the corresponding left PLN serves as the control for the PLN from the injected hind foot.
  • Percent suppression is calculated using the non-drug treated animals as allogenic control. Rapamycin at a dose of 6 mg kg, p.o. gave 86% suppression, whereas cyclosporin A at the same dose gave 43% suppression.
  • Results are expressed by the following ratio: 3 H-PLN cells control C3H mouse - 3 H-PLN cells rapamvcin-treated C3H mouse 3 H-PLN cells control C3H mouse - 3 H-PLN cells test compound-treated C3H mouse
  • the second in vivo test procedure is designed to determine the survival time of pinch skin graft from male DBA/2 donors transplanted to male BALB/c recipients.
  • the method is adapted from Billingham R.E. and Medawar P.B., J. Exp. Biol. 28:385- 402, (1951). Briefly, a pinch skin graft from the donor is grafted on the dorsum of the recipient as a homograft, and an autograft is used as control in the same region.
  • the recipients are treated with either varying concentrations of cyclosporin A as test control or the test compound, intraperitoneally. Untreated recipients serve as rejection control.
  • the graft is monitored daily and observations are recorded until the graft becomes dry and forms a blackened scab. This is considered as the rejection day.
  • the mean graft survival time (number of days ⁇ S.D.) of the drug treatment group is compared with the control group.
  • Antifungal activity of the compounds of this invention was measured against 5 strains of Candida albicans using a plate test procedure for measurement of inhibition. The following represents the typical procedure used. Compound to be tested was placed on sterile dried 1/4" plate disks, and allowed to dry. Agar plates were seeded with fungi and allowed to solidify. The impregnated disks were placed on the seeded Agar surface and incubated for the time required for the particular culture. Results are expressed in MIC ( ⁇ g/ml) to inhibit growth. The results of this test procedure showed that the compounds of this invention have antifungal activity; however, it was surprising that the compounds of this invention were less active than the parent compound, rapamycin.
  • the compounds are useful in the treatment of transplantation rejection such as, heart, kidney, liver, bone marrow, and skin transplants; autoimmune diseases such as, lupus, rheumatoid arthritis, diabetes mellitus, myasthenia gravis, and multiple sclerosis; and diseases of inflammation such as, psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease; and fungal infections.
  • transplantation rejection such as, heart, kidney, liver, bone marrow, and skin transplants
  • autoimmune diseases such as, lupus, rheumatoid arthritis, diabetes mellitus, myasthenia gravis, and multiple sclerosis
  • diseases of inflammation such as, psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease
  • fungal infections e.g., osis, dermatitis, eczema, seborrhea,
  • 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.
  • the carrier is a finely divided solid which is in admixture with the finely divided active ingredient.
  • 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).
  • the carrier can also be an oily ester such as ethyl oleate and isopropyl myristate.
  • Sterile liquid carriers are useful in sterile liquid form compositions for parenteral administration.
  • the liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellent.
  • Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by, for example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously. The compound can also be administered orally either in liquid or solid composition form.
  • the pharmaceutical composition is in unit dosage form, e.g. as tablets or capsules.
  • the composition is sub-divided in unit dose containing appropriate quantities of the active ingredient;
  • the unit dosage forms can be packaged compositions, for example, packeted powders, vials, ampoules, prefilled syringes or sachets containing liquids.
  • the unit dosage form can be, for example, a capsule or tablet itself, or it can be the appropriate number of any such compositions in package form.
  • the dosage to be used in the treatment must be subjectively determined by the attending physician.
  • the compounds of this invention may be employed as a solution, cream, or lotion by formulation with pharmaceutically acceptable vehicles containing 0.1- 5 percent, preferably 2%, of active compound which may be administered to a fungally affected area.
  • the following representative compounds can be prepared from rapamycin and the appropriately terminally-N-substituted amino diacid monoester by employing the method used to prepare the title compound in Example 5.
  • Example 11 HPLC analysis showed that the monoester is a 7.5 :1 mixture of two conformers.
  • the title compound was prepared (0.83 g, 12.8%) along with the 42- monoester as described in Example 10. HPLC analysis showed that the diester is a 7.7:1 mixture of two conformers.
  • Rapamycin-14,31,42-tris (monomethylsuccinate) Rapamycin-14,31,42-tris (monophenyl-3',3'-dimethylglutarate)
  • Rapamycin-14,31,42-tris (mono t-butyl-3'-methylglutarate)
  • Rapamycin-14,31,42-tris (monobenzylthiodiglycolate)
  • Rapamycin-14,31,42-tris (monohexyldiglycolate)
  • Rapamycin-14,31,42-tris (monopropylphthalate) Rapamycin-14,31,42-tris (monoethyl-2',6'-pyridinedicarboxylate) Example 15.
  • Rapamycin-31 ,42-bis (monomethylsuccinate) Rapamycin-31 ,42-bis (monophenyl-3',3'-dimethylglutarate)
  • Rapamycin-31,42-bis (mono t-butyl-3'-methylglutarate)
  • Rapamycin-31 ,42-bis (monopropylphthalate) Rapamycin-31,42-bis (monoethyl-2',6'-pyridinedicarboxylate)
  • the crude product was purified via reverse phase HPLC on a Cig column eluting starting with 60 % acetonitrile/water. Collected, after, concentration, 586 mg (24 %) of rapamycin-31, 42-bishemiglutarate. J H NMR (CDCI3.
  • Rapamycin-31 ,42-bishemi-3'-methylglutarate Rapamycin-31 ,42-bishemi-3',3'-dimethylglutarate Rapamycin-31 ,42-bishemi-3'-oxoglutarate
  • ion FAB 1112 (M-l, free acid), 994, 589, 475, 297, 167, 148, 117, 99 (100); High Res. MS (neg. ion FAB) Calcd for C6iH 8 9 ⁇ 9 NNa (M-Na) 1162.5926, Found 1162.5899.
  • the crude product was purified via reverse phase HPLC on a Cis column eluting starting with 60 % acetonitrile/water. After concentration, 870 mg ( 26 %) of rapamycin-42-hemi-3'-oxoglutarate and 500 mg (13 %) of rapamycin-31,42-bishemi-3'oxoglutarate were isolated.
  • the following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 20.
  • the crude product was purified via reverse phase HPLC on a C ⁇ 8 column gradient eluting starting with 20 % acetonitrile/water to 60 % acetonitrile/water. Collected, after, concentration, 770 mg (31 %) of rapamycin-31,42-bishemisuccinate.
  • the purified bis-31,42 hemisuccinate of rapamycin (770 mg, 686 umol) was dissolved in 10 mL of 95 % ethanol and 166 mg (1.37 mmol) of tris(hydroxymethyl)- methylamine was added. Water (1 mL) was added to completely dissolve the amine. Once dissolved, the yellow solution was concentrated in vacuo to give a foamy yellow solid. The very hygroscopic foam was dried in a drying pistol for 24 h, refluxing over acetone at reduced pressure to give 890 mg (95 %) of the bistromethamine salt. The bistromethane salt was evaluated in the standard pharmacological test procedures.

Abstract

L'invention se rapporte à un composé représenté par la structure (I); où: R1, R2 et R3 représentent chacun séparément hydrogène ou R4; R4 représente (a), (b) ou (c); R5 représente hydrogène, alkyle, aralkyle, -(CH2)qCO2R8, -(CH2)rNR9CO2R10, carbamylalkyle, aminoalkyle, hydroxyalkyle, guanylalkyle, mercaptoalkyle, alkylthioalkyle, indolylméthyle, hydroxyphénylméthyle, imidazoylméthyle, ou phényle, éventuellement monosubstitué, disubstitué ou trisubstitué par un substituant choisi parmi alkyle, alkoxy, hydroxy, cyano, halo, nitro, carbalkoxy, trifluorométhyle, amino ou un acide carboxylique, R6 et R9 représentent chacun séparément hydrogène, alkyle ou aralkyle; R7, R8 et R10 représente chacun séparément alkyle, aralkyle, fluorénylméthyle ou phényle, éventuellement monosubstitué, disubstitué ou trisubstitué; R11 et R12 représentent chacun séparément alkyle, aralkyle ou phényle, éventuellement monosubstitué, disubstitué ou trisubstitué; X représente (d), O ou S; R13 et R14 représentent chacun séparément hydrogène ou alkyle; Y représente C, H ou N;; m est compris entre 0 et 4; n est compris entre 0 et 4; p est compris entre 1 et 2; q est compris entre 0 et 4; r est compris entre 0 et 4; t est compris entre 0 et 4; et u est compris entre 0 et 4; R5, R6, m et n étant indépendants dans chacune des sous-unités (e), lorsque p est égal à 2; ou à un sel pharmaceutiquement acceptable du composé de la structure (I), à condition que R1, R2 et R3 ne représentent pas tous hydrogène, à condition que R1, R2 et R3 ne représentent pas tous (a), et à condition en outre que t et u ne soient pas tous deux égaux à 0, lorsque X représente O ou S. Grâce à son action immunosuppressive, un tel composé est utile dans le traitement des phénomènes de rejet de la greffe, dans les affections dues aux réactions de l'hôte contre le greffon, dans les maladies auto-immunes et dans les maladies inflammatoires et grâce à son action fongicide, un tel composé est utile dans le traitement des mycoses.The invention relates to a compound represented by structure (I); where: R1, R2 and R3 each independently represent hydrogen or R4; R4 represents (a), (b) or (c); R5 represents hydrogen, alkyl, aralkyl, - (CH2) qCO2R8, - (CH2) rNR9CO2R10, carbamylalkyl, aminoalkyl, hydroxyalkyl, guanylalkyl, mercaptoalkyl, alkylthioalkyl, indolylmethyl, hydroxyphenylmethyl, imidazoylmethyl, disubstituted, or disubstituted, or disubstituted, or disubstituted selected from alkyl, alkoxy, hydroxy, cyano, halo, nitro, carbalkoxy, trifluoromethyl, amino or a carboxylic acid, R6 and R9 each independently represent hydrogen, alkyl or aralkyl; R7, R8 and R10 each independently represents alkyl, aralkyl, fluorenylmethyl or phenyl, optionally monosubstituted, disubstituted or trisubstituted; R11 and R12 each independently represent alkyl, aralkyl or phenyl, optionally monosubstituted, disubstituted or trisubstituted; X represents (d), O or S; R13 and R14 each independently represent hydrogen or alkyl; Y represents C, H or N ;; m is between 0 and 4; n is between 0 and 4; p is between 1 and 2; q is between 0 and 4; r is between 0 and 4; t is between 0 and 4; and u is between 0 and 4; R5, R6, m and n being independent in each of the subunits (e), when p is equal to 2; or to a pharmaceutically acceptable salt of the compound of structure (I), provided that R1, R2 and R3 do not all represent hydrogen, provided that R1, R2 and R3 do not all represent (a), and further provided that t and u are not both equal to 0, when X represents O or S. Thanks to its immunosuppressive action, such a compound is useful in the treatment of the phenomena of rejection of the graft, in the affections due to the reactions of the host against the graft, in autoimmune diseases and in inflammatory diseases and thanks to its fungicidal action, such a compound is useful in the treatment of yeast infections.

Description

CARBOXY TC ACTD ESTERS OF RAPAMYCTN
BACKGROUND OF THE INVENTION
This invention relates to novel esters of rapamycin and a method for using them in the treatment of transplantation rejection, host vs. graft disease, autoimmune diseases, diseases of inflammation, and fungal infections.
Rapamycin is a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus. which was found to have antifungal activity, particularly against Candida albicans. both in vitro and in vivo [C. Vezina et al., J. Antibiot. 28, 721 (1975); S.N. Seghal et al., J. Antibiot. 28, 727 (1975); H. A. Baker et al., J. Antibiot. 31, 539 (1978); U.S. Patent 3,929,992; and U.S. Patent 3,993,749].
Rapamycin alone (U.S. Patent 4,885,171) or in combination with picibanil (U.S. Patent 4,401,653) has been shown to have antitumor activity. R. Mattel et al. [Can. J. Physiol. Pharmacol. 55, 48 (1977)] disclosed that rapamycin is effective in the experimental allergic encephalomyelitis model, a model for multiple sclerosis; in the adjuvant arthritis model, a model for rheumatoid arthritis; and effectively inhibited the formation of IgE-like antibodies.
The immunosuppressive effects of rapamycin have been disclosed in FASEB 3, 3411 (1989), rapamycin has been shown to be effective in inhibiting transplant rejection (U.S. Patent Application Ser. No. 362,544 filed June 6, 1989). Cyclosporin A and FK-506, other macrocyclic molecules, also have been shown to be effective as immunosuppressive agents, therefore useful in preventing transplant rejection [FASEB 3, 3411 (1989); FASEB 3, 5256 (1989); and R. Y. Calne et al., Lancet 1183 (1978)]. Mono- and diacylated derivatives of rapamycin (esterified at the 28 and 43 positions) have been shown to be useful as antifungal agents (U.S. Patent 4,316,885) and used to make water soluble prodrugs of rapamycin (U.S. Patent 4,650,803). Recently, the numbering convention for rapamycin has been changed; therefore according to Chemical Abstracts nomenclature, the esters described above would be at the 31- and 42- positions. DESCRIPTION OF THE INVENTION
This invention provides derivatives of rapamycin which are useful as immunosuppressive, anti-inflammatory, and antifungal agents having the structure
wherein R1, R2> and R3 are each, independently, hydrogen, or R4;
O
H ?
R4 is -[C(CH2)niCH(CH2)nN]pC02R7 , -C- (CH2)tX(CH2)uCO2R11 , or
I I
R5 R6
R5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms,
-(CH2)qCO2R8, -(CH2)rNR9CO2R10, carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazolylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R6 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms; R7, R8, and R10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri- substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R1 ! and R12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R13 and R14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms; Y is CH or N; m is 0 - 4; n is 0 - 4; p is 1 - 2 q is 0 - 4 r is 0 - 4; t is O - 4; u is 0 - 4;
O wherein R >5 , R , , and n are independent in each of the[C(CH2)mCH(CH2)nN]
R5 R6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R1, R2* and R3 are not all hydrogen, further provided that R1, R2- and R3 are not all
O
II
— [C(CH2)mCH(CH2)nN]pCO2R7 , and still further provided that t and u are not
I I
R5 R6 both 0 when X is O or S. O
II
Of the compounds when R4 is — [C(CH2)mCH(CH2)-.N]pCO2R7
I I
R5 R6 preferred members are those in which m = 0, n = 0, and p = l; m = 0, n = 0, and p = 2; n = 0, and R5 is -(CH2)qCO2R8; m = 0, n = 0, and R5 is -(CH2)rNR9CO2R10; and m = 0, n = 0, and R5 is hydrogen. Preferred compounds also include those
O
Δ. 'I 11 members in which R4 is -C- (CH2)tX(CH2)uCO2Rn .
The pharmaceutically acceptable salts may be formed from inorganic cations such as sodium, potassium, and the like; mono-, di-, and trialkyl amines of 1-6 carbon atoms, per alkyl group and mono-, di-, and trihydroxyalkyl amines of 1-6 carbon atoms per alkyl group; and organic acids such as acetic, lactic, citric, tartaric, succinic, maleic, malonic, gluconic, and the like. Preferred basic salts are formed from sodium cations and tris(hydroxymethyl)methyl-.mine.
The compounds of this invention can be prepared by acylating rapamycin with an acylating agent having the general structures
, Z- VC-(CH2)tX(CH2)uCO2Rn , or
where Z is OH in the presence of a coupling reagent, such as dicyclohexyl- carbodiimide. The compounds of this invention also can be prepared using an anhydride or a mixed anhydride of the above described carboxylic acid as the acylating species. Alternatively, the acylating species can be an acid halide, where Z can be Cl, Br, or I. The acylating groups used to prepare the compounds of this invention are commercially available or can be prepared by methods that are disclosed in the literature.
Where it is desired to prepare acyl derivatives having two or three different R4 groups then sequential acylation may be performed using appropriate acylating agents as defined above, if necessary isolating the desired product by appropriate purification techniques. In general the 42-position is acylated first and such a monoacylated product may be isolated prior to the second acylation and so forth. Appropriate protecting groups may be used to block any position where acylation is not required.
Immunosuppressive activity was evaluated in an in vitro standard pharmacological test procedure to measure lymphocyte proliferation (LAF) and in two in vivo standard pharmacological test procedures. The first in vivo procedure was a popliteal lymph node (PLN) test procedure which measured the effect of compounds of this invention on a mixed lymphocyte reaction and the second in vivo procedure evaluated the survival time of a pinch skin graft. The comitogen-induced thymocyte proliferation procedure (LAF) was used as an in vitro measure of the immunosuppressive effects of representative compounds. Briefly, cells from the thymus of normal B ALB/c mice are cultured for 72 hours with PHA and IL-1 and pulsed with tritiated thymidine during the last six hours. Cells are cultured with and without various concentrations of rapamycin, cyclosporin A, or test compound. Cells are harvested and incorporated; radioactivity is determined. Inhibition of lymphoproliferation is assessed in percent change in counts per minute from non-drug treated controls. The results are expressed by the following ratio, or as the percent inhibition of lymphoproliferation of 1 μM. 3H-control thvmus cells - H3-rapamvcin-treated thvmus cells
3H-control thymus cells - H3-test compound-treated cells
A mixed lymphocyte reaction (MLR) occurs when lymphoid cells from genetically distinct animals are combined in tissue culture. Each stimulates the other to undergo blast transformation which results in increased DNA synthesis that can be quantified by the incorporation of tritiated thymidine. Since stimulating a MLR is a function of disparity at Major Histocompatibility antigens, an in vivo popliteal lymph node (PLN) test procedure closely correlates to host vs. graft disease. Briefly, irradiated spleen cells from B ALB/c donors are injected into the right hind foot pad of recipient C3H mice. The drug is given daily, p.o. from Day 0 to Day 4. On Day 3 and Day 4, tritiated thymidine is given i.p., b.i.d. On Day 5, the hind popliteal lymph nodes are removed and dissolved, and radioactivity counted. The corresponding left PLN serves as the control for the PLN from the injected hind foot. Percent suppression is calculated using the non-drug treated animals as allogenic control. Rapamycin at a dose of 6 mg kg, p.o. gave 86% suppression, whereas cyclosporin A at the same dose gave 43% suppression. Results are expressed by the following ratio: 3H-PLN cells control C3H mouse - 3H-PLN cells rapamvcin-treated C3H mouse 3H-PLN cells control C3H mouse - 3H-PLN cells test compound-treated C3H mouse
The second in vivo test procedure is designed to determine the survival time of pinch skin graft from male DBA/2 donors transplanted to male BALB/c recipients. The method is adapted from Billingham R.E. and Medawar P.B., J. Exp. Biol. 28:385- 402, (1951). Briefly, a pinch skin graft from the donor is grafted on the dorsum of the recipient as a homograft, and an autograft is used as control in the same region. The recipients are treated with either varying concentrations of cyclosporin A as test control or the test compound, intraperitoneally. Untreated recipients serve as rejection control. The graft is monitored daily and observations are recorded until the graft becomes dry and forms a blackened scab. This is considered as the rejection day. The mean graft survival time (number of days ± S.D.) of the drug treatment group is compared with the control group.
The following table summarizes the results of representative compounds of this invention in these three standard test procedures.
TABLE 1
TABLE 1 (Continued)
Compound Example 16 Example 17 Example 18 Example 19 Example 20 Example 21 Example 22 Example 23 Example 24 Rapamycin
** Result expressed as percent inhibition of lymphoproliferation at 1 μM. + Not evaluated ++ Results obtained using cremophore/ethanol as a vechicle for administration. Ratios of 0.33 and 1.07 were also obtained using carboxymethyl cellulose as a vehicle for administration. ## Results obtained using cremophore/ethanol as a vechicle for administration. Ratios of 0.20 and 1.08 also were obtained using carboxymethyl cellulose as a vehicle for administration. ### A ratio of 0.42 also was obtained for this compound.
The results of these standard pharmacological test procedures demonstrate immunosuppressive activity both in vitro and in vivo for the compounds of this invention. Positive ratios in the LAF and PLN test procedures indicate suppression of T cell proliferation. As a transplanted pinch skin grafts are typically rejected within 6-7 days without the use of an immunosuppressive agent, the increased survival time of the skin graft when treated with the compounds of this invention further demonstrates their utility as immunosuppressive agents. While it appears that the compound disclosed by Examples 12 and 21 may cause T cell proliferation in the PLN test procedure, it is believed a negative ratio in this test procedure coupled with an increased survival time observed in the skin graft test procedure indicates a proliferation of TSUppressor cells, which are implicated in suppressing the immune response, (see, I. Roitt et al. Immunology, CV.Moseby Co. 1989, p 12.8-12.11).
Antifungal activity of the compounds of this invention was measured against 5 strains of Candida albicans using a plate test procedure for measurement of inhibition. The following represents the typical procedure used. Compound to be tested was placed on sterile dried 1/4" plate disks, and allowed to dry. Agar plates were seeded with fungi and allowed to solidify. The impregnated disks were placed on the seeded Agar surface and incubated for the time required for the particular culture. Results are expressed in MIC ( μg/ml) to inhibit growth. The results of this test procedure showed that the compounds of this invention have antifungal activity; however, it was surprising that the compounds of this invention were less active than the parent compound, rapamycin.
Table 2*
Strain of Candida albicans
Table 2* (Continued) Strain of Candida albicans
* expressed as MIC (μg/ml) + not evaluated
Based on the results of these standard pharmacological test procedures, the compounds are useful in the treatment of transplantation rejection such as, heart, kidney, liver, bone marrow, and skin transplants; autoimmune diseases such as, lupus, rheumatoid arthritis, diabetes mellitus, myasthenia gravis, and multiple sclerosis; and diseases of inflammation such as, psoriasis, dermatitis, eczema, seborrhea, inflammatory bowel disease; and fungal infections. The compounds may be administered neat or with a pharmaceutical carrier to a mammal in need thereof. The pharmaceutical carrier may be solid or liquid.
A solid carrier can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in admixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of the active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.
Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid carrier 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 propellent.
Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by, for example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously. The compound can also be administered orally either in liquid or solid composition form.
Preferably, the pharmaceutical composition is in unit dosage form, e.g. as tablets or capsules. In such form, the composition is sub-divided in unit dose containing appropriate quantities of the active ingredient; the unit dosage forms can be packaged compositions, for example, packeted powders, vials, ampoules, prefilled syringes or sachets containing liquids. The unit dosage form can be, for example, a capsule or tablet itself, or it can be the appropriate number of any such compositions in package form. The dosage to be used in the treatment must be subjectively determined by the attending physician. In addition, the compounds of this invention may be employed as a solution, cream, or lotion by formulation with pharmaceutically acceptable vehicles containing 0.1- 5 percent, preferably 2%, of active compound which may be administered to a fungally affected area.
The following examples illustrate the preparation of representative compounds of this invention. Example 1
Rapamycin-42-ester with N-fH . l-dimethylethoxy)carbonyl l-glycylglycine
Under anhydrous conditions, a solution of rapamycin (3 g, 3.28 mmole) and
N-[(l,l-dimethylethoxy)carbonyl]-glycylglycine (3.04 g, 13.1 mmole) in 40 mL of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide (1.35 g, 6.56 mmole) followed by 4-dimethylaminopyridine (0.8 g, 6.56 mmole). After stirring at ambient temperature for 48 hours, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were absorbed directly onto silica gel Merck 60 by adding the gel and evaporation to dryness. Flash chromatography of the preabsorbed material (using a gradient elution with ethylacetate-toluene from 2:1 to 1:0 v/v) afforded 1.05 g (28.3 %) of the title compound isolated as a three quarter toluene solvate, along with the 31,42-diester of Example 2. HPLC analysis showed that the monoester is a 8.3: 1 mixture of two conformers. H NMR (CDCI3, 400 MHz): δ 1.46 (m, 9H, COOBu1), 1.654 (s, 3H,
CH3C=C), 1.751 (s, 3H, CH3C=C), 3.14 (s, 3H, CH3O), 3.33 (s, 3H, CH3O), 3.36 (s, 3H, CH3O), 4.18 (d, 1H, CtfOH), 4.75 (m, 1H, 42-CHO ), 4.79 (s, 1H, OH); High Res. MS (neg. ion FAB) Calcd for C60H93N3O1 : 1127.6504, measured mass 1127.6474.
Anal. Calcd for C60H93N3O17 • 0.75 PI1CH3: C, 65.45; H, 8.33; N, 3.51
Found: C, 65,23; H, 8.32; N, 3.86
The following representative compounds can be prepared from rapamycin and the appropriate terminally-N-substituted amino acid by employing the method used to prepare the title compound in Example 1.
Rapamycin-42-ester with N-[(fluorenylmethoxy)carbonyl]-alanylserine Rapamycin-42-ester with N-[(fluorenylmethoxy)carbonyl]-glycylglycine Rapamycin-42-ester with N-[(ethoxy)carbonyl]-arginylmethionine Rapamycin-42-ester with N-[(4l-chlorophenoxy)carbonyl]-histidylarginine Rapamycin-42-ester with N-[(phenoxy)carbonyl]-tryptophanylleucine Rapamycin-42-ester with N-[(phenylmethoxy)carbonyl)]-N-methylglycyl-N- ethylalanine Rapamycin-42-ester with N-[(phenylmethoxy)carbonyl]-N-methyl-β- alanylphenylalanine
Rapamycin-42-ester with N-[(l,l-d-methylethoxy)carbonyl]-cysteinylglycine
Example 2
Rapamydn-31.42-diesterwith N-r(l.l-dimethylethoxy')carbonyl1-glycylglycine
The title compound (1.85 g, 42%) was separated from the 42-monoester as described in Example 1 and isolated as a three quarter toluene solvate. HPLC analysis showed that the diester is a 8.1: 1 mixture of conformers.
!H NMR (CDCI3, 400 MHz): δ 1.452 (m, 18H, COOBu*), 1.6612 (s, 3H, CH3C=C), 1.7815 (s, 3H, CH3C=C), 3.14 (s, 3H, OCH3), 3.34 (s, 3H, OCH3), 3.35 (s, 3H, OCH3), 4.52 (s, IH, OH), 4.79 (m, IH, 42- ΑO ); High Res. MS (neg. ion FAB): Calcd for C69H107N5O21 1341.7458, measured mass: 1341.7463.
Anal. Calcd for C69H107N5O21 • 0.75 PI1CH3: C, 63.17; H, 8.06; N, 4.96 Found: C, 62.83; H, 8.09; N, 5.00
The following representative compounds can be prepared from rapamycin and the appropriate terminally-N-substituted amino acid by employing the method used to prepare the title compound in Example 2.
Rapamycin-31 ,42-diester with N-[(fluorenylmethoxy)carbonyl]-alanylserine Rapamycin-31.42-diester with N-[(fluorenylmethoxy)carbonyl]-glycylglycine Rapamycin-31,42-diester with N-[(ethoxy)carbonyl]-arginylmethionine Rapamycin-31,42-diester with N-[(4'-chlorophenoxy)c-trbonyl]-histidylarginine Rapamycin-31,42-diester with N-[(phenoxy)carbonyl]-tryptophanylleucine Rapamycin-31,42-diester with N-[(phenylmethoxy)carbonyl)]-N-methylglycyl-N- ethyl-alanine
Rapamycin-31,42-diester with N-[(phenylmethoxy)carbonyl]-N-methyl-β- alanylphenyl- alanine Rapamycin-31 ,42-diester with N-[( 1 , 1 -dimethylethoxy)carbonyl] -cysteinylglycine Example 3
Rapamycin-31 ,42-diester with N-IY1.1 -dimethylethoxy)carbonyl1-N-methylglycine
Under anhydrous conditions, an ice cold solution of rapamycin (2 g, 2.18 mmole) and Nα-Boc sarcosine (1.65 g, 8.75 mmole) in 20 ml of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide (1.8 g, 8.7 mmole) followed by 4-dimethylaminopyridine (1 g, 8.7 mmole). After stirring overnight at ambient temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were evaporated to dryness to give an amorphous amber solid (3 g). The crude product was purified by flash chromatography ( on silica Merck 60, elution with hexane-ethylacetate 1:1, v/v) to provide the title compound (0.75 g, 27.4%) along with the 42-monoester of Example 4. HPLC analysis showed that the diester is a 19.8:1 mixture of two conformers. The multiplicity of the NMR peaks suggests the presence of amide rotamers.
IH NMR (CDC13, 400 MHz): δ 1.411, 1.438, 1.448 and 1.474 (m, 18 H, COOBut), 2.91 (m, 6H, NCH3), 3.14 (s, 3H, CH3O), 3.34 (s, 3H, CH3O), 3.37 (s, 3H, CH3O), 4.73 (broad, IH, 42-CHO), 4.82 (2s, IH, OH); High Res. MS (neg. ion FAB): Calcd. for C67H105N3O19 1255.7342, measured mass 1255.7289. Anal. Calcd for C67H105N3O19: C, 64.04; H, 8.42; N, 3.34
Found: C, 64.14; H, 8.74; N, 3.63
The following representative compounds can be prepared from rapamycin and the appropriate terminally-N-substituted amino acid by employing the method used to prepare the title compound in Example 3.
Rapamycin-31,42-diester with N-[(ethoxy)carbonyl]-tyrosine
Rapamycin-31 ,42-diester with N-[(fluorenylmethoxy)carbonyl]-phenylalanine
Rapamycin-31 ,42-diester with N-[(3',4',5'-trihydroxyphenoxy)carbonyl]-isoleucine Rapamycin-31,42-diester with N-[(l,l-dimethylethoxy)carbonyl)-glutamine Rapamycin-31,42-diester with N-[(phenoxy)carbonyl]-N-methylalanine Rapamycin-31,42-diester with N-[(propyloxy)carbonyl]-4-aminobutryic acid Rapamycin-31,42-diester with N-[(phenylmethoxy)carbonyl]-7-aminoheptanoic acid Rapamycin-31,42-diester with N-[(fluorenylmethoxy)carbonyl]-serine Example 4
Rapamycin-42-esterwith N-r(l.l-dimethylethoxy')carbonyn-N-methylglvcine
Under anhydrous conditions, an ice cold solution of rapamycin (0.95 g, 1.02 mmole) and Nα-Boc sarcosine (0.21 g, 1.1 mmole) in 20 mL of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide 0.21 g, 1 mmole) followed by 4-dimethylaminopyridine (0.12 g, 1 mmole). After stirring for 4 hours at ambient temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were concentrated in vacuo to give an amorphous amber solid. Flash chromatography of the crude product (on silica Merck 60, elution with hexane- ethylacetate 1:1 v/v to remove the diester of Example 3, followed by chloroform- ethylacetate-methanol 75:25:1 v/v) provided partially purified title compound (0.38 g, 35%). Pure product was obtained by preparative HPLC (Waters Prep 500, silica gel, chloroforai-ethylacetate-methanol 75:25: 1 v/v, flow rate 250 mL min). HPLC analysis showed that the ester is a 6.6:1 mixture of two conformers. The multiplicity of NMR peaks suggests the presence of amide rotamers.
IH NMR (CDCI3, 400 MHz): δ 1.42-1.46 (ds, 9H, COOBu*), 2.91 (ds, 3H, NCH3), 1.644 (s, 3H, CH3C=C), 1.738 (s, 3H, CH3C=C), 3.12 (s, 3H, CH3O), 3.32 (s, 3H, CH3O), 3.35 (s. 3H, CH3O), 4.18 (d, IH, CHOH), 4.71 (broad, IH, 42-CHO), 4.78 (broad s, IH, OH); High Res. MS (neg. ion FAB): Calcd for C59H92N2O16 1084.6446, measured mass 1084.6503.
Anal. Calcd for C59H92N2O16- C, 65.29; H, 8.54; N, 2.58 Found: C, 65.25; H, 8.52; N, 2.42
The following representative compounds can be prepared from rapamycin and the appropriate terminally-N-substituted amino acid by employing the method used to prepare the title compound in Example 4.
Rapamycin-42-ester with N-[(ethoxy)carbonyl]-tyrosine
Rapamycin-42-ester with N-[(fluorenylmethoxy)carbonyl]-phenylalanine Rapamycin-42-ester with N-[(3',4',5'-trihydroxyphenoxy)carbonyl]-isoleucine Rapamycin-42-ester with N-[(l,l-dimethylethoxy)carbonyl)-glutamine Rapamycin-42-ester with N-[(phenoxy)carbonyl]-N-methylalanine Rapamycin-42-ester with N-[(propyloxy)carbonyl]-4-aminobutryic acid
Rapamycin-42-ester with N-[(phenylmethoxy)carbonyl]-7-aminoheptanoic acid Rapamycin-31,42-diester with N-[(fluorenylmethoxy)carbonyl]serine
Example 5
Rapamycin-31.42-diester with 5-(l.l-dimethylethoxy)-2-rr(l.l-dimethylethoxy)- carbonynaminol-5-oxopentanoic acid
Under anhydrous conditions, an ice cold solution of rapamycin (4 g, 4.37 mmole) and L-glutamic acid Nα~Boc-γ-tert-butylester (4.9 g, 16.1 mmole) in 40 mL of dry dichloromethane was treated with dicyclohexylcarbodiimide (1.8 g, 8.7 mmole) followed by 4-dimethylaminopyridine (1 g, 8.7 mmole). After stirring overnight at room temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were concentrated in vacuo to provide 11 g of an amorphous amber solid. The crude product was purified by flash chromatography (on silica Merck 60, gradient elution with hexane-ethylacetate from 2:1 to 1:1, v/v) to yield 4.52 g (69.6%) of the title compound along with the 42-monoester of Example 6. HPLC analysis showed that the diester consists of a 6.6: 1 mixture of two conformers. H NMR (CDCI3, 400 MHz): δ 1.42 (m, 36 H, COOBu1), 1.646 (s, 3H,
CH3C=C), 1.701 (s, 3H, CH3C=C), 3.13 (s, 3H, CH3O), 3.34 (s, 3H, CH3O), 3.36 (s, 3H, CH3O), 4.735 (m, 2H, OH+42-CH-O); High Res. MS (neg. ion FAB): calc. for C79H125N3O23 1483.8715, measured mass 1483.8714.
Anal. Calcd for C79H125N3O23: C, 63.90; H, 8.49; N, 2.83 Found: C, 63.63 H, 8.41; N, 2.44
The following representative compounds can be prepared from rapamycin and the appropriately terminally-N-substituted amino diacid monoester by employing the method used to prepare the title compound in Example 5.
Rapamycin-31,42-diester with 6-(phenylmethoxy)-2-[[fluorenylmethoxy)carbonyl]- amino]-6-oxohexanoic acid
Rapamycin-31 ,42-diester with 6-(4'-methylphenoxy)-3-[[(phenylmethoxy)carbonyl]- amino-6-oxohexanoic acid
Rapamycin-31,42-diester with 6-(ethoxy)-4-[[(phenoxy)carrx)nyl]amino]-6-oxo- hexanoic acid Rapamycin-31,42-diester with 6-(methoxy)-5-[[(ethoxy)carbonyl]amino]-6-oxo- hexanoic acid
Rapamycin-31,42-diester with 4-(phenoxy)-2-[N-[(l,l-dimethylethoxy)carbonyl]-N- methylamino]-4-oxobutanoic acid Rapamycin-31,42-diester with 4-(phenylmethoxy)-3-[N-[(methoxy)carbonyl]-N- ιι_ethylamino]-4-oxobutanoic acid
Example 6
Rapamycin-42-ester with 5-(l . l-dimethylethoxy)-2-rr(l . l-dimethylethoxyV carbonynaminol-5-oxopentanoic acid
The title compound (1.14 g, 20.6%) was separated from the 31,42-diester as described in Example 5 and isolated as the quarter hydrate/mono-ethyl acetate solvate. HPLC analysis showed that the monoester is a 11.5: 1 mixture of two conformers.
!H NMR (CDCI3, 400 MHz): δ 1.425 (m, 18H, COOBu1), 1.643 (s, 3H,
CH3C=C), 1.737 (s, 3H, CH3C=C), 3.13 (s, 3H, CH3O), 3.32 (s, 3H, CH3O),
3.36 (s, 3H, CH3O), 4.17 (d, IH, CHOH), 4.71 (M, IH, 42-CHO), 4.785 (s, IH, OH); High Resolution MS ( neg. ion FAB): Calc. for C65Hχo2N2θι 1198.7127, measured mass 1198.7077.
Anal. Calcd for C65H102N2O18 • CH3COOEt • 0.25 H2O: C, 64.13, H, 8.60; N, 2.17
Found: C, 64.18; H, 8.52: N, 2.01
The following representative compounds can be prepared from rapamycin and the appropriately terminally-N-substituted amino diacid monoester by employing the method used to prepare the title compound in Example 6.
Rapamycin-42-esterwit 6-(phenylmethoxy)-2-[[flucM«nylmethoxy)c--rbonyl]-a-i-ino]- 6-oxohexanoic acid
Rapamycin-42-ester with 6-(4'-methylphenoxy)-3-[[(phenylmethoxy)carbonyl]-amino- 6-oxohexanoic acid Rapamycin-42-iester with 6-(ethoxy)-4-[[(phenoxy)carbonyl]amino]-6-oxo- hexanoic acid
Rapamycin-42-ester with 6-(methoxy)-5-[[(ethoxy)carbonyl]amino]-6-oxo- hexanoic acid Rapamycin-42-ester with 4-(phenoxy)-2-[N-[(l,l-dimethylethoxy)carbonyl]-N- methylamino]-4-oxobutanoic acid
Rapamycin-42-ester with 4-(phenylmethoxy)-3-[N-[(methoxy)carbonyl]-N- me ylainino]-4-oxobutanoic acid
Example 7
Rapamycin-31.42-diester with 2-rr(l.l-dimethylethoxy')carbonynamino1-4-oxo-4- (phenylmethoxy) butanoic acid
Under anhydrous conditions, 295mg (1.21mmol) of 2,4,6 trichlorobenzoyl chloride was added to a solution of 391mg(1.21mmol) of Nα-Boc-L-aspartic acid-β- benzyl ester and 170μL (1.21mmol) of Et3N in 1 mL of THF at room temperature. After stirring for 30 minutes, 500 mg (0.55mmol) of rapamycin and 295 mg ( 2.42 mmol) of dimethylaminopyridine was added and the reaction was left to stir overnight. The reaction mixture was then filtered and the filtrate concentrated in vacuo. Pure product (200 mg, 25%) was obtained by preparative HPLC (5 cm column, 40 % ethyl acetate-hexane). The product was isolated as the heptahydrate.
*H NMR (CDC13, 400 MHz) δ 7.347 (s, 10 H, Ar), 6.223, 5.126 (s, 4 H, CH2Ph), 4.698 (m, 1 H, CH-CO2), 4.587 (m, 2 H, NH), 3.353 (s, 3 H, CH3O), 3.337 (s, 3
H, CH3O), 3.301 (s, 3 H, CH3O), 2.775 (m, 4 H, CH2C02); IR (KBr) 3420 (OH),
2935 (CH), 2920 (CH), 1730 (C=O), 1650, 1500, 1455, 1370, 1170 cm"1; MS (neg. ion FAB) 1523 {Mr), 1433, 297, 248, 205, 148, 44, 25 (100).
Anal. Calcd for C83H1 ι7N3θ23-7H20 C, 60.40; H, 7.09; N, 2.54 Found: C, 60.54; H, 7.28; N, 2.56 Example 8
Rapamycin-31.42-diester with 3-rrfLl-dimethylethoxy)carbonyllamino1-4-oxo-4- (phenylmethoxy. butanoic acid
Under anhydrous conditions, 532 mg (2.18 mmol) of 2,4,6 trichlorobenzoyl chloride in 1 mL THF was added to a solution of 704 mg (2.18 mmol) of Nα-Boc-L- aspartic acid-α-benzyl ester and 303 μL (2.18 mmol) of Et3N in 5 mL of THF at room temperature. After stirring for 20 minutes, the reaction mixture was filtered over sintered glass, and the precipitate was washed with THF. The filtrate was concentrated in vacua to give a thick oil. The oil was dissolved in 5 mL of benzene and 1.00 g (1.09 mmol) of rapamycin and 532 mg (4.36 mmol) of dimethylaminopyridine in 1 mL of benzene was added dropwise. The reaction was stirred for 2 hr, poured into ethyl acetate, and washed consecutively with 0.5 N HC1 and brine. The solution was dried over sodium sulfate, decanted, concentrated in vacua to give a white foamy solid, which was purified via flash chromatography on a 60 mm x 100 mm silica column (20- 40 % ethyl acetate/hexane as eluant) to give 532 mg (33 %) of the title compound which was isolated as the hydrate.
!H NMR (CDC13, 400 MHz) δ 7.362 (s, 10 H, Ar), 5.193 (s, 4 H, CH2Ph), 4.596 (m, 1 Η, CH-CO2), 4.586 (m, 2 Η, NH), 3.336 (s, 3 Η, CH5O), 3.306 (s, 3 Η, CH5O), 3.145 (s, 3 Η, CH3O); IR (KBr) 3410 (OΗ), 2950 (CΗ), 2920 (CΗ), 1735 (C=0), 1710 (C=0), 1640, 1490, 1445, 1350, 1150 cm -1; MS (neg. ion FAB) 1524 (M-), 1434, 297, 248, 232, 214, 205, 167, 148, 42 (100), 26.
Anal. Calcd for C83Η117Ν3O23 • H2O: C, 65.38; H, 7.73; N, 2.76 Found: C, 64.85; H, 7.67; N, 2.56
Example 9
Rapamvcin-42-ester with 3-rKl .l-dimethylethoxy)carbonyl1amino]-4-oxo-4- (phenylmethoxy) butanoic acid
The title compound (374 mg, 23%) was prepared by the method described in the previous Example and separated from the compound described in the previous Example by flash chromatography (20-40% ethyl acetate/hexane as the eluant) and isolated as the sesquihydrate. *H NMR (CDCI3, 400 MHz) δ 7.356 (s, 5 H, Ar), 5.185 (s, 2H, CH2Ph),
4.635 (m, 1 Η, CH-CO2), 4.582 (m, 1 Η, NH), 3.330 (s, 6Η, CH3O), 3.135 (s, 3
Η, CH3O); IR (KBr) 3410 (OΗ), 2950 (CΗ), 2920 (CΗ), 1735 (C=O), 1710 (C=O),
1640, 1490, 1445, 1350, 1150cm-1; MS (neg. ionFAB) 1218 (M-), 1127, 590, 168, 42, 25, 17 (100).
Anal. Calcd for Q57Η98Ν2O18 • 1.5 H2O: C, 63.64; H, 8.21; N, 2.22 Found: C, 63.64; H, 7.51; N, 2.13
Example 10
Rapamycin-42-ester with 5-(l.l-dimethyloxy)-4-rr(l.l-dimethylethoxy carbonyn aminol-5-oxopentanoic acid
Under anhydrous conditions, an ice cold solution of rapamycin (4 g, 4.37 mmole) and L-glutamic acid Nα-Boc-α-tert-butylester (4.9 g, 16.1 mmole) in 40 mL of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide (1.8 g, 8.7 mmole) followed by 4-dimethylamino pyridine (1 g, 8.7 mmole). After stirring overnight at ambient temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were concentrated in vacua to give 9 g of an amorphous amber solid. The crude product was purified by flash chromatography (on silica Merck 60, gradient elution with hexane-ethylacetate from 2:1 to 3:2, v/v) to provide 1.35 g (25.7%) of the title compound along with the 31,42-diester of
Example 11. HPLC analysis showed that the monoester is a 7.5 :1 mixture of two conformers.
JH NMR (CDCI3, 400 MHz): δ 1.43 (s, 9H, COOBu1) and 1.46 (s, 9H, COOBut), 1.65 (s, 3H, CH3C=C), 1.75 (s, 3H, CH3C=C), 3.14 (s, 3H, CH3O), 3.34 (s, 3H, CH3O), 3.38 (s, 3H, CH3O), 4.18 (d, IH, CH-OH), 4.65 (m, IH, 42- CHO), 4.80 (s, IH, OH);
High Res. MS (neg. ion FAB): Calc. for C65H102N2O18. 1198.7126, measured mass 1198.7135.
Anal. Calcd for C65Hι02N2θι8: C, 65.09; H, 8.57; N, 2.34
Found C, 65.04; H, 8.33; N, 2.64 Example 11
Rapamvcin-31.42-diester with 5-(l.l-dimethylethoxyV4-rr(l.l-dimethylethox .- carbonyll- aminol-5-oxopentanoic acid
The title compound was prepared (0.83 g, 12.8%) along with the 42- monoester as described in Example 10. HPLC analysis showed that the diester is a 7.7:1 mixture of two conformers.
*H NMR (CDCI3, 400 MHz): δ 1.43 (s, 18H, COOBu*), 1.46 (s, 18H,
COOBu1), 1.659 (s, 3H, CH3C=C), 1.759 (s, 3H, CH3C=C), 3.14 (s, 3H, CH3O),
3.34 (s, 3H, CH3O), 3.38 (s, 3H, CH3O), 4.66 (m, IH, 42-CHO), 4.72 (s, IH,
OH); High Res. MS (neg. ion FAB): Calcd for C79H125N3O23: 1483.8704, measured mass 1483.8636.
Anal. Calcd for C79H125N3O23: C, 63.90; H, 8.49; N, 2.83 Found: C, 63.68; H, 8.60; N, 3.20
Example 12
Rapamvcin-42-ester with N-3-. NS-bisr(l.l-dimethylethoxy)carbonyn-L-lvsine
Under anhydrous conditions, a solution of rapamycin (3 g, 3.28 mmole) and
Nα, ISP-bis-Boc-L-lysine (4.5 g, 13 mmole) in 40 mL of anhydrous dichloromethane was treated with dicyclohexylcarbodiimide (1.35 g, 6.56 mmole) followed by 4- dimethylaminopyridine (0.8 g, 6.56 m mole). After stirring overnight at ambient temperature, the precipitated solid was collected and washed with dichloromethane. The combined filtrates were concentrated in vacuo to give an amorphous amber solid. Flash chromatography of the crude product (on silica Merck 60, elution with hexane- ethylacetate 1:1 v/v) gave partially purified title compound. Pure product (0.8 g, 19.6%) was obtained by preparative HPLC (Waters Prep 500, silica gel, hexane- ethylacetate 3:2 v/v, flow rate 250 mL min). HPLC analysis showed that the monoester is a 9: 1 mixture of two conformers. !H NMR (CDCI3, 400 MHz): δ 1.438 (m, 9H, COOBu1), 1.455 (s, 9H, COOBut), 652 (S, 3H, CH3C=C), 1.752 (s, 3H, CH3C=C), 3.14 (s, 3H, CH3O), 3.33(s, 3H, CH3O), 3.37 (s, 3H, CH3O), 4.18 (d, IH, CHOH), 4.72 (m, IH, 42- CHO), 4.79 (s, IH, OH); High Res. MS (neg. ion FAB): Calcd for C67Hκ)7N3θi8:
1241.7549, measured mass 1241.7604.
Anal. Calcd for C67H107N3O18 : C, 64.76; H, 8.68; N, 3.38
Found: C, 64.58; H, 9.01; N, 3.10
Example 13
Rapamycin-31.42-diester with N&. N----bisr(l.l-dimethylethoxy)c_trbonyl.-L-lvsine
Under a nitrogen atmosphere, a solution of Nα,Nε bis-Boc-L-lysine (1.038 g, 3 mmole) and triethylamine (0.42 mL, 3 mmmole) in 10 mL of anhydrous THF was treated in one portion with 2,4,6-trichlorobenzoyl chloride (0.73 g, 3 mmole). After stirring for 20 minutes at ambient temperature, the precipitated solid was collected and the filtrate was concentrated in vacuo . The resulting mixed anhydride was dissolved in 5 mL of benzene and added to a stirred solution of rapamycin (1 g, 1.09 mmole) containing 4-dimethylamino pyridine (0.59 g, 4.8 mmole) in 10 mL of benzene. After stirring at ambient temperature overnight, the precipitated solid was collected and the filtrate was evaporated to dryness (yellow foam). The crude product was purified by flash chromatography ( on silica Merck 60, elution with hexane-ethylacetate 1:1) to provide title compound (1.15 g, 67%). HPLC analysis shows that the diester is a 9:1 mixture of two conformers.
IH NMR (CDCI3, 400 MHz): δ 1.426 (m, 9H, COOBut), 1.438 (s, 9H, COOBut),
1.443 (s, 9H, COOBut), 1.446 (s, 9H, COOBut), 3.141 (s, 3H, CH3O), 3.36 (s, 3H,
CH3O), 3.378 (s, 3H, CH3O), 4.68-4.76 (m, 2H, OH and 42-CHO); High res. MS
(neg. ion FAB): Calcd. for C83H135N5O23 1569.9526, measured mass 1569.9537. Anal. Calcd. for C83H135N5O23: C, 63.46; H, 8.66; N, 4.46
Found: C, 63.06; H, 8.84; N, 4.09 Example 14.
Rapamycin- 14.31 -42-tris(monobenzylsuccinate)
To a solution of 5.0 g (5.47 mmol) of rapamycin, 3.41 g (16.41 mmol) of monobenzylsuccinate, and 3.15 g (16.41 mmol) of l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride in 20 mL of dry dichloromethane was added 200 mg of 4-dimethylaminopyridine. The solution was stirred at room temperature for 3 days. The reaction mixture was poured into 2 N HCl and extracted three times with ethyl acetate. The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a light yellow foam. Hash chromatography on a 60 mm x 150 mm silica gel column eluting with 20 % ethyl acetate/hexane to 75 % ethyl acetate/hexane gave three fractions. Fraction #1, upon concentration, gave 330 mg (4.1 %) of pure rapamycin- 14,31, 42-tris- (monobenzylsuccinate).
*H NMR (CDC13, 400 MHz) δ 7.353 (bs, 15 H, arom), 5.168 (d, J = 2.0 Hz, 1 H, CH-O2C), 5.148 (m, 6 Η, CH2Ph), 4.672 (m, 1 Η, CO2CH-CΗOMe), 3.355 (s, 3 H, CH5O-), 3.337 (s, 3 Η, CH3O-), 3.327 (s, 3 Η, CH3O-), 2.697 ( m, 12 Η, θ2CCH2CH2Cθ2CΗ2Ph), 1.745 (s, 3 H, CH3C=C), 1.655 (s, 3 H, CH3C=C); IR (KBr) 3450 (OH), 2950 (CH), 1745 (C=0), 1650, 1460, 1385, 1360, 1160, 1105, 995 cm-1.
Analysis Calcd for C84Hi09NO2ι - 3 H20 C 66.27; H 7.56; N 0.92 Found C 65.96; H 7.24; N 1.00
The following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 14.
Rapamycin-14,31,42-tris (monomethylsuccinate) Rapamycin-14,31,42-tris (monophenyl-3',3'-dimethylglutarate)
Rapamycin-14,31,42-tris (mono t-butyl-3'-methylglutarate)
Rapamycin-14,31,42-tris (monobenzylthiodiglycolate)
Rapamycin-14,31,42-tris (monohexyldiglycolate)
Rapamycin-14,31,42-tris (monopropylphthalate) Rapamycin-14,31,42-tris (monoethyl-2',6'-pyridinedicarboxylate) Example 15.
Rapamycin-31.42-bis(monobenzylsuccinate)
Fraction # 2, obtained from the procedure employed in Example 14, gave
1.25 g (17.7 %) of pure rapamycin-31,42-bis(monobenzylsuccinate) upon concen¬ tration. lH NMR (CDC13, 400 MHz) δ 7.351 (bs, 10 H, arom). 5.168 (d, J = 2.0 Hz, 1 H, CH-O2C), 5.125 (m, 4 Η, CH2Ph), 4.680 (m, 1 Η, CO2CH-CΗOMe), 3.356 (s, 3 H, CH3O-), 3.329 (s, 3 H, CH3O-), 3.146 (s, 3 Η, CH3O-), 2.639 ( m, 8 Η, O2CCH2CH2CO2CΗ2P--), 1.748 (s, 3 H, CHiC=C), 1.654 (s, 3 Η, CH3C=C); IR (KBr) 3450 (OΗ), 2940 (CΗ), 1740 (C=O), 1650, 1455, 1380, 1355, 1160, 1105, 995 cm-1; MS (neg. ion FAB) 1294 (M-), 1202, 1103, 1012, 590, 511, 475, 297, 207, 167, 148, 99 (100); High Res. MS (neg. ion FAB) Calcd for C73H99NO19 1293.68108, found 1293.6811.
Analysis Calcd for C73H99NO19 Η2O C 66.82; H 7.70; N 1.07
Found C 67.17; H 7.67; N 1.23
The following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 15.
Rapamycin-31 ,42-bis (monomethylsuccinate) Rapamycin-31 ,42-bis (monophenyl-3',3'-dimethylglutarate)
Rapamycin-31,42-bis (mono t-butyl-3'-methylglutarate)
Rapamycin-31 ,42-bis (monobenzylthiodiglycolate)
Rapamycin-31 ,42-bis (monohexyldiglycolate)
Rapamycin-31 ,42-bis (monopropylphthalate) Rapamycin-31,42-bis (monoethyl-2',6'-pyridinedicarboxylate)
Example 16.
Rapamycin-42-(monobenzylsuccinate)
Fraction # 3, obtained from the procedure employed in Example 14, gave 930 mg (15.4 %) of pure rapamycin-42-monobenzylsuccinate upon concentration. iH NMR (CDCI3, 400 MHz) δ 7.355 (bs, 5 H, arom), 5.141 (m, 2 H, CH2Ph), 4.680 (m, 1 Η, CC^CH-CΗOMe), 3.364 (s, 3 H, CH3O-), 3.333 (s, 3 Η, CH3O-), 3.141 (s, 3 Η, CH5O-), 2.698 ( m, 4 Η, O2CCH2CH2CO2CΗ2PI1), 1.751 (s, 3 H, CH3C=C), 1.655 (s, 3 Η, CH3C=C); IR (KBr) 3450 (OΗ), 2940 (CΗ), 1740 (C=0), 1645, 1455, 1380, 1165, 1105, 990 cm-1; MS (neg. ion FAB) 1103 (M-), 1045, 1012, 624, 590, 167, 99 (100); High Res. MS (neg. ion FAB) Calcd for C 2H89NO16 1103.6181, found 1103.6048.
Analysis Calcd for C62H89NOi6 - H20 C 66.36; H 8.02; N 1.24
Found C 66.02; H 7.69; N 1.26
The following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 16.
Rapamycin-42-(monomethylsuccinate)
Rapamycin-42-monophenyl-3',3'-dimethylglutarate)
Rapamycin-42-(mono t-butyl-3'-methylglutarate)
Rapamycin-42-(monobenzylthiodiglycolate)
Rapamycin-42-(monohexyldiglycolate) Rapamycin-42-(monopropylphthalate)
Rapamycin-42-(monoethyl-2,,6'-pyridinedicarboxylate)
Example 17.
Rapamycin-31.42-bishemi lutarate
To a solution of 2.0 g (2.2 mmol) of rapamycin in 10 mL of dry dichloromethane was added 1.24 g (10.9 mmol) of glutaric anhydride followed by 881 uL (861 mg, 10.9 mmol) of pyridine. To this was added 200 mg of 4-dimethylaminopyridine and the reaction mixture was allowed to reflux for 8 h. The solution was cooled to room temperature, poured into 2 N HCl, and extracted three times with dichloromethane. The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow foam. The crude product was purified via reverse phase HPLC on a Cig column eluting starting with 60 % acetonitrile/water. Collected, after, concentration, 586 mg (24 %) of rapamycin-31, 42-bishemiglutarate. JH NMR (CDCI3. 400 MHz) δ 5.398 (m, 1 H, -Cθ2CHCHOMe), 4.683 (m, 1 H, -CO2CHCHOMe), 3.364 (s, 3 H, CH30-), 3.362 (s, 3 H, CH3O-), 3.106 (s, 3 H, CH3O-), 2.407 (m, 8 H, -O2CCH2CH2CH2CO2H), 1.960 (m, 4 H, -O2CCH2CH2CH2CO2H), 1.770 (s, 3 H, CH3C--C), 1.653 (s, 3 H, CH3C=C); 1 C NMR (CDCI3, MHz) 211.45 (C=O), 206.84 (C=O), 200.44 (C=O), 177.83 (C=0), 177.04 (C=0), 172.43 (C=0), 171.20 (C=0), 165.27 (C=0), 159.08 (C=0); IR (KBr) 3430 (OH), 2940 (CH), 2880 (CH), 1745 (C=O), 1685, 1625, 1580, 1450, 1385, 1330, 1200, 1140, 1100, 990 cm-1; MS (neg. ion FAB) 1140 (M-H), 1122, 1026, 990, 946, 913, 590, 475, 435, 321, 167, 148, 131 (100), 113; High Res. MS (neg. ion FAB) Calcd for C6ιH9oOι9N (M-H) 1140.6107, Found 1140.6106. Analysis Calcd for Q51H91O19N • H2O C 63.15; H 8.02; N 1.20
Found C 63.35; H 7.88; N 1.40
The following representative compounds can be prepared from rapamycin and the appropriate anhydride by employing the method used to prepare the title compound in Example 17.
Rapamycin-31 ,42-bishemi-3'-methylglutarate Rapamycin-31 ,42-bishemi-3',3'-dimethylglutarate Rapamycin-31 ,42-bishemi-3'-oxoglutarate
Rapamycin-31,42-bishemi-3'-thioglutarate Rapamycin-31 ,42-bishemi-phthalate Rapamycin-31 ,42-bishemi-2',3'-pyridine dicarboxylate
Example 18.
Rapamycin-31.42-hemiglutarate bissodium salt
Purified bis-31,42-hemiglutarate of rapamycin (740 mg, 649 umol), prepared as described in Example 17, was dissolved in 5 mL of 95 % ethanol and 107 mg (1.27 mmol) of sodium bicarbonate was added. Water (1 mL) was added to completely dissolve the salt. Once dissolved, the light yellow solution was concentrated in vacuo to give a foamy yellow solid. The foam was dried in a drying pistol for 24 h, refluxing over acetone at reduced pressure to give 520 mg of the bissodium salt. *H NMR (d6-DMSO, 400 MHz) δ 5.235 (m, 1 H, -CHO2C), 4.498 (m, 1 Η, MeOCΗCHO2C-), 3.287 (s, 6 Η, 2 CH3O-), 3.236 (s, 3 Η, CH3O-), 2.245 (m, 8 Η, O2CCH2CΗ2CH2CO2-), 1.712 (s, 3 Η, CH3C=C), 1.593 (s, 3 Η, CH3C=C); IR (KBr) 3420 (OΗ), 2920 (CΗ), 1725 (C=O), 1675, 1620, 1560, 1450, 1400, 1375, 1230, 1195, 1130, 1090, 980 cm"1; MS (neg. ion FAB) 1112 (M-l, free acid), 994, 589, 475, 297, 167, 148, 117, 99 (100); High Res. MS (neg. ion FAB) Calcd for C6iH89θι9NNa (M-Na) 1162.5926, Found 1162.5899.
Analysis Calcd for C6iHs9θi9NNa2 - H2θ C 60.85; H 7.56; N 1.16 Found C 60.67; H 7.36; N 1.58
Example 19.
Rapamycin-31.42-bishemiglutarate bistromethamine salt
Purified bis-31,42 hemiglutarate of rapamycin (950 mg, 833 umol), prepared as described in Example 17, was dissolved in 5 mL of 95 % ethanol and 197 mg (1.63 mmol) of tris(hydroxymethyl)methylamine was added. Water (1 mL) was added to completely dissolve the amine. Once dissolved, the yellow solution was concentrated in vacuo to give a foamy yellow solid. The very hygroscopic foam was dried in a drying pistol for 24 h, refluxing over acetone at reduced pressure to give 900 mg (78 %) of the bistromethamine salt.
lH NMR (d6-DMSO, 400 MHz) δ 5.253 (m, 1 H, -CHO2C), 4.523 (m, 1 Η,
MeOCΗCH02C-), 3.347 (s, 6 Η, 2 CH3O-), 3.276 (s, 3 Η, CH3O-), 2.289 (m, 8 Η, O2CCH2CΗ2CH2CO2-), 1.681 (s, 3 Η, CH3C=C), 1.595 (s, 3 Η, CH3C=C); IR (KBr) 3400 (OΗ), 2920 (CΗ), 1730 (C=O), 1620, 1555, 1450, 1400, 1370, 1185, 1060, 980 cm-1; MS (neg. ion FAB) 1140 (M-Η, free acid), 1028, 167, 148, 131 (100), 113; High Res. MS (neg. ion FAB) Calcd for C 1H90O19N (M-H, free acid) 1140.6107, Found 1140.6069.
Analysis Calcd for C69Hιo3θ25N3 - 2 H2O C 58.77; H 7.58; N 2.98 Found C 58.47; H 7.94; N 3.58 Example 20.
Rapamvcin-42-hemi-3'-oxoglutarate
To a solution of 3.0 g (3.3 mmol) of rapamycin in 20 mL of dry dichloromethane was added 1.90 g (16.4 mmol) of diglycolic anhydride followed by 1.32 mL (1.29 g, 16.4 mmol) of pyridine. To this was added 200 mg of 4-dimethylaminopyridine and the reaction mixture was allowed to stir at room temperature for 2 days. The solution was cooled to room temperature, poured into 2 N HCl, and extracted three times with dichloromethane. The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow foam. The crude product was purified via reverse phase HPLC on a Cis column eluting starting with 60 % acetonitrile/water. After concentration, 870 mg ( 26 %) of rapamycin-42-hemi-3'-oxoglutarate and 500 mg (13 %) of rapamycin-31,42-bishemi-3'oxoglutarate were isolated. l NMR (CDC13, 400 MHz) δ 4.768 (m, 1 H, CO2CH-CHOMe), 4.250 (m, 4 H, O2CCH2OCH2CO2), 3.356 (s, 3 Η, CH3O-), 3.331 (s, 3 Η, CH3O-), 3.139 (s, 3 Η, CH3 -), 1.759 (s, 3 Η, CH3C--C), 1.653 (s, 3 Η, CH3C=C); IR (KBr) 3420 (OΗ), 2920 (CΗ), 2875 (CΗ), 1740 (C=O), 1720 (C=O), 1640, 1625, 1445, 1370, 1320, 1200, 1135, 1095, 980 cm-1; MS (neg. ion FAB) 1028 (M - Η), 327, 167 (100), 148, 133, 115; High Res. MS (neg. ion FAB) Calcd for C55H82O17N (M - H) 1028.5597, Found 1028.5599.
Analysis Calcd for C55H83θι7N - 3 H2O C 60.97; H 8.22; N 1.29 Found C 61.33; H 7.74; N 1.69
The following representative compounds can be prepared from rapamycin and the appropriate half acid-ester by employing the method used to prepare the title compound in Example 20.
Rapamycin-42-hemi-3'-methylglutarate
Rapamycin-42-hemi-3l,3'-dimethylglutarate Rapamycin-42-hemi-3'-thioglutarate Rapamycin-42-hemi-phthalate Rapamycin-42-hemi-2',3,-pyridine dicarboxylate Example 21.
Rapamycin-31.42-bishemi-3'-oxoglutarate
To a solution of 5.0 g (5.47 mmol) of rapamycin in 20 mL of dry dichloromethane was added 3.17 g (27.3 mmol) of diglycolic anhydride followed by 2.17 mL (2.12 g, 27.3 mmol) of pyridine. To this was added 400 mg of 4-dimethylaminopyridine and the reaction mixture was allowed to stir at reflux for 24 h. The solution was cooled to room temperature, poured into 2 N HCl, and extracted three times with dichloromethane. The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow foam. The crude product was purified via reverse phase HPLC on a Ci8 column eluting starting with 60 % acetonitrile/water. After concentration, 1.75 g ( 28 %) of rapamycin-31,42-bishemi-3'-oxoglutarate was isolated. *H NMR (CDC13, 400 MHz) δ 4.785 (m, 1 H, CO2CHCHOMe), 4.260
(m, 8 H, O2CCH2OCH2CO2), 3.360 (s, 3 Η, CH3O-), 3.343 (s, 3 Η, CH3O-), 3.143 (s, 3 Η, CH3O-), 1.775 (s, 3 Η, CH3C=C), 1.656 (s, 3 Η, CH?C=C); 13C NMR (CDCI3, MHz) 211.12 (C=O), 207.73 (C=O), 193.11 (C=O), 171.90 (C=O), 171.59 (C=O), 170.15 (C=O), 169.35 (C=O), 168.83 (C=O), 166.63 (C=O); IR (KBr) 3420 (OH), 2920 (CH), 2850 (CH), 1740 (C=0), 1645, 1625, 1440, 1370, 1190, 11300, 980 cm"1; MS (neg. ion FAB) 1140 (M-H), 1122, 1026, 990, 946, 913, 590, 475, 435, 321, 167, 148, 131 (100), 113; High Res. MS (neg. ion FAB) Calcd for C59H86O21N (M - H) 1144.5701, Found 1144.5702.
Analysis Calcd for C59H87O21N C 61.82; H 7.65; N 1.22 Found C 61.59; H 7.36; N 1.84
Example 22.
Rapamycin-31.42-bishemi-3'-oxoglutarate disodium salt
Purified bis-31,42 hemi-3'-oxoglutarate of rapamycin (720 mg, 629 umol) , prepared by the procedure employed in Example 21, was dissolved in 10 mL of 95 % ethanol and 106 mg (1.26 mmol) of sodium bicarbonate was added. Water (1 mL) was added to completely dissolve the salt. Once dissolved, the light yellow solution was concentrated in vacuo to give a foamy yellow solid. The foam was dried in a drying pistol for 48 h, refluxing over dichloromethane at reduced pressure to give 435 mg (58 %) of the disodium salt.
*H NMR (d6-DMSO, 400 MHz) δ 4.975 (m, 1 H, -CH02C), 4.593 (m, 1 H, MeOCHCHO2C-), 4.135 (s, 2 Η, -O2CCΗ2OCH2CO2R), 3.617 (s,2 Η, O2CCH2OCΗ2CO2R), 3.299 (s, 6 H, 2 CH3O-), 3.232 (s, 3 Η, CH3O-), 1.614
(s, 3 Η, CH3C=C), 1.553 (s, 3 Η, CH3C=C); IR (KBr) 3420 (OΗ), 2920 (CΗ), 1735 (C=O), 1615, 1445, 1395, 1380, 1320, 1220, 1130, 1090, 980 cm"1; MS (neg. ion FAB) 1188 (M-l), 1166 (M-Na), 1144, 1051, 1028, 590, 459, 167, 155 (100), 148, 133, 115. Analysis Calcd for C59Η85θ2iNNa2 • 2Η2O C 57.79; H 7.26; N 1.14
Found C 57.94; H 7.11; N 1.26
Example 23.
Raparnycin-31.42-bishemi-3'-oxoglutarate bistromethamine salt
Purified bis-31,42 hemi-3'-oxoglutarate of rapamycin (1.01 g, 882 umol), prepared by the procedure employed in Example 21, was dissolved in 10 mL of 95 % ethanol and 213 mg (1.76 mmol) of tris(hydroxymethyl)- methylamine was added. Water (1 mL) was added to completely dissolve the amine. Once dissolved, the yellow solution was concentrated in vacuo to give a foamy yellow solid. The very hygroscopic foam was dried in a drying pistol for 48 h, refluxing over dichloromethane at reduced pressure to give 805 mg (66 %) of the bistromethamine salt.
*H NMR (d6-DMSO, 400 MHz) δ 4.955 (m, 1 H, -CHO2Q, 4.600 (m, 1 H, MeOCHCHO2C-), 4.149 (s, 2 Η, -O2CCΗ2OCH2CO2R), 3.770 (s, 2 Η, -O2CCH2OCΗ2CO2R), 3.407 (s, 6 H, 2 CH3O-), 3.257 (s, 3 Η, CH3O-), 1.806 (s, 3 Η, CH3C=C), 1.614 (s, 3 Η, CH3C=C); IR (KBr) 3400 (OΗ), 2920 (CΗ), 1730 (C=O), 1620, 1550, 1450, 1395, 1370, 1200, 1060, 985 cm"1; MS (neg. ion FAB) 1144 (M-Η, free acid), 1028, 167, 148, 133 (100), 115.
Analysis Calcd for C67Η109O27N3 • Η20 C 57.22; H 7.90; N 2.98 Found C 57.26; H 7.90; N 3.15 Example 24.
Rapamycin-31.42-bishemisuccinate.
To a solution of 2.0 g (2.2 mmol) of rapamycin in 10 mL of dry dichloro¬ methane was added 1.19 g (10.9 mmol) of succinic anhydride followed by 881 uL (861 mg, 10.9 mmol) of pyridine. To this was added 200 mg of 4-dimethylamino- pyridine and the reaction mixture refluxed for 24 h. The solution was cooled to room temperature, poured into 2 N HCl, and extracted three times with dichloromethane. The combined organic extracts were washed with brine, dried over anhydrous sodium sulfate, decanted, and concentrated in vacuo to give a yellow foam. The crude product was purified via reverse phase HPLC on a Cχ8 column gradient eluting starting with 20 % acetonitrile/water to 60 % acetonitrile/water. Collected, after, concentration, 770 mg (31 %) of rapamycin-31,42-bishemisuccinate.
The purified bis-31,42 hemisuccinate of rapamycin (770 mg, 686 umol) was dissolved in 10 mL of 95 % ethanol and 166 mg (1.37 mmol) of tris(hydroxymethyl)- methylamine was added. Water (1 mL) was added to completely dissolve the amine. Once dissolved, the yellow solution was concentrated in vacuo to give a foamy yellow solid. The very hygroscopic foam was dried in a drying pistol for 24 h, refluxing over acetone at reduced pressure to give 890 mg (95 %) of the bistromethamine salt. The bistromethane salt was evaluated in the standard pharmacological test procedures.
*H NMR (d6-DMSO, 400 MHz) 5.231 (m, 1 H, -CHO2C), 4.554 (m, 1 Η, MeOCΗCHO2C-), 3.426 (s, 6 Η, 2 CH3O-), 3.249 (s, 3 Η, CH3O-), 2.431 (m, 8 Η, O2CCΗ2CΗ2CO2-), 1.700 (s, 3 H, CH3C=C), 1.554 (s, 3 Η, CH3C=C); 13C NMR (d6-DMSO, ) 211.28 (C=0), 205.23 (C=0), 199.59 (C=0), 174.86 (C=0), 173.62 (C=O), 171.72 (C=O), 171.50 (C=0), 166.56 (C=O), 166.53 (C=0); IR (KBr) 3420 (OΗ), 2940 (CΗ), 1735 (C=0), 1630, 1580, 1460, 1400, 1380, 1170, 1070, 990 cm-1; MS (neg. ion FAB) 1112 (M-l, free acid), 994, 589, 475, 297, 167, 148, 117, 99 (100).
Analysis Calcd for C 7Ηιo9θ25N3 - 2 Η2O C 57.80; H 8.12; N 3.01 Found C 57.91; H 8.21; N 2.37

Claims

CLAIMSWhat is claimed is:
1. A compound of the structure
wherein R1, R2> and R3 are each, independently, hydrogen, or R4;
O
II O II
R4 is [C(CH2)mCH(CH2)nN]pCO2R7 -C- (CH2)tX(CH2)uCO2Rπ or
I I
R5 R6
R5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms,
-(CH2)qC02R8, -(CH2)rNR9Cθ2R10, carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazoylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R6 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms;
R7, R8, and R10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri- substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R11 and R12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R13
X is -C-, O, or S;
I R14
R13 and R14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms;
Y is CH orN; m is 0 - 4; n is 0 - 4; p is 1 - 2; q is 0 - 4; r is 0 - 4; tis O - 4; u is 0 - 4;
O
II wherein R5, R6, m, and n are independent in each of the[C(CH2)mCH(CH2)nN]
I I
R5 R6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R1, R2> and R3 are not all hydrogen, further provided that R1, R2> and R3 are not all O
II
- [C(CH2)mCH(CH2)nN]pCO2R7 , and still further provided that t and u are not
I I
R5 R6 both 0 when X is O or S.
O
II
2. A compound of claim 1 where R4 is — [C(CH2)mCH(CH2)nN]pCO2R7
I I
R5 R6 m = 0, n = 0, and p = 1 or a pharmaceutically acceptable salt thereof.
O
II
3. A compound of claim 1 where R4 is - [C(CH2)rnCH(CH2)nN]pCO2R7
I I
R5 R6 m = 0, n = 0, and p = 2 or a pharmaceutically acceptable salt thereof.
O
II
4. A compound of claim 1 where R4 is — [C(CH2)mCH(CH2)nN]pCO2R7
I I R5 R6 n = 0, and R5 is -(CH2)qCθ2R8 or a pharmaceutically acceptable salt thereof.
O
II
5. A compound of claim 1 where R4 is - [C(CH2)mCH(CH2)nN]pC02R7
I I
R5 R6 m = 0, n = 0, and R5 is -(CH2)rNR9CO2R10 or a pharmaceutically acceptable salt thereof.
O
II
6. A compound of claim 1 where R4 is - [C(CH2)mCH(CH2)nN]pC02R7
I I
R5 R6 m = 0, n = 0, and R5 is hydrogen or a pharmaceutically acceptable salt thereof. O
__ " 11
7. A compound of claim 1 where R4 is -C- (CH2)tX(CH2)uC02Rn or a pharmaceutically acceptable salt thereof.
8. A compound of claim 1 which is rapamycin-42-ester with N-[(l,l-dimethyleth- oxy)carbonyl]-glycylglycine or a pharmaceutically acceptable salt thereof.
9. A compound of claim 1 which is rapamycin-31,42-diester with N-[(l,l-di- methyl- ethoxy)carbonyl]-glycylglycine or a pharmaceutically acceptable salt thereof.
10. A compound of claim 1 which is rapamycin-31,42-diester with N-[(l,l-di- methylethoxy)carbonyl]-N-methylglyc_ne or a pharmaceutically acceptable salt thereof.
11. A compound of claim 1 which is rapamycin-42-ester with N-[(l,l-di- methylethoxy)carbonyl]-N-methylglycine or a pharmaceutically acceptable salt thereof.
12. A compound of claim 1 which is rap amycin-31,42-diester with 5-(l,l- dimethylethoxy)-2-[[(l,l-dimethylethoxy)carbonyl]amino]-5-oxopentanoic acid or a pharmaceutically acceptable salt thereof.
13. A compound of claim 1 which is rapamycin-42-ester with 5-(l,l- dimethylethoxy)-2-[[(l,l-dimethylethoxy)carbonyl]amino]-5-oxopentanoic acid or a pharmaceutically acceptable salt thereof.
14. A compound of claim 1 which is rapamycin-31,42-diester with 2-[[(l,l- dimethylethoxy)carbonyl]amino]-4-oxo-4-(phenylmethoxy) butanoic acid or a pharma¬ ceutically acceptable salt thereof.
15. A compound of claim 1 which is rapamycin-31,42-diester with 3- [[(l,l-dimethylethoxy)carbonyl]amino]-4-oxo-4-(phenylmethoxy) butanoic acid or a pharmaceutically acceptable salt thereof.
16. A compound of claim 1 which is rapamycin-42-ester with 3- [[(l,l-dimethylethoxy)carbonyl]amino]-4-oxo-4-(phenylmethoxy) butanoic acid or a pharmaceutically acceptable salt thereof.
17. A compound of claim 1 which is rapamycin-42-ester with 5-(l,l-dimethyl- oxy)-4-[[(l,l-dimethylethoxy)carbonyl]amino]-5-oxopentanoic acid or a pharma¬ ceutically acceptable salt thereof.
18. A compound of claim 1 which is rapamycin-31,42-diester with 5-(l,l- dimethylethoxy)-4-[[(l,l-dimethylethoxy)carbonyl]amino]-5-oxopentanoic acid or a pharmaceutically acceptable salt thereof.
19. A compound of claim 1 which is rapamycin-42-ester with Nα, Nε-bis[(l,l- dimethylethoxy)carbonyl]-L-lysine or a pharmaceutically acceptable salt thereof.
20. A compound of claim 1 which is rapamycin-31,42-diester with Nα, Nε bis[(l,l-dimethylethoxy)carbonyl]-L-lysine or a pharmaceutically acceptable salt thereof.
21. A compound of claim 1 which is rapamycin- 14,31, 42-tris(monobenzyl- succinate) or a pharmaceutically acceptable salt thereof.
22. A compound of claim 1 which is rapamycin-31 ,42-bis(monobenzylsuccinate) or a pharmaceutically acceptable salt thereof.
23. A compound of claim 1 which is rapamycin-42-(monobenzylsuccinate) or a pharmaceutically acceptable salt thereof.
24. A compound of claim 1 which is rapamycin-31, 42-bishemiglutarate or a pharmaceutically acceptable salt thereof.
25. A compound of claim 1 which is rapamycin-31,42-hemiglutarate bissodium salt.
26. A compound of claim 1 which is rapamycin-31, 42-bishemiglutarate bistromethamine salt
27. A compound of claim 1 which is rapamycin-42-hemi-3'-oxoglutarate or a pharmaceutically acceptable salt thereof.
28. A compound of claim 1 which is rapamycin-31 ,42-bishemi-3'-oxoglutarate or a pharmaceutically acceptable salt thereof.
29. A compound of claim 1 which is rapamycin-31,42-bishemi-3'-oxoglutarate disodium salt.
30. A compound of claim 1 which is rapamycin-31,42-bishemi-3'-oxoglutarate bistromethamine salt.
31. A compound of claim 1 which is rapamycin-31,42-bishemisuccinate or a pharmaceutically acceptable salt thereof.
32. A compound of claim 1 which is rapamycin-31,42-bishemisuccinate bistromethane salt
33. A method of treating transplantation rejection, host vs. graft disease, autoimmune diseases, and diseases of inflammation in a mammal by administering an immunosuppressive amount of a compound having the structure
wherein R1, R2> and R3 are each, independently, hydrogen, or R4; o
II O
II
R4 is - [C(CH2)mCH(CH2)nN]pCO2R7 , -C- (CH2)tX(CH2)uCO2Rπ , or
I I
R5 R6
R5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, -(CH2)qC02R8, -(CH2)rNR9C02R10, carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazoylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R6 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms; R7, R8, and R10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri- substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R11 and R12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
ϊ13 X is -C-, O, or S;
I R14
R13 and R14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms; Y is CH or N; m is 0 - 4; n is 0 - 4; p is 1 - 2; q is 0 - 4; r is 0 - 4; tis O - 4; u is 0 - 4;
O wherein R »5 , r R«6°, m, and n are independent in each of the[C(CH2)mCH(CH2)nN]
I I
R5 R6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R1, R2> and R3 are not all hydrogen, further provided that R1, R2- and R3 are not all
O
II
- [C(CH2)mCH(CH2)nN]pCO2R7 , and still further provided that t and u are not
I I
R5 R6 both 0 when X is O or S.
34. A method of treating fungal infections which comprises administering an antifungal amount of a compound having the structure
wherein R1, R2> and R3 are each, independently, hydrogen, or R4; O
II °
R4 is -[C(CH2)mCH(CH2)nN]pC02R7 , -C-(CH2)tX(CH2)uC02R11 , or
I I
R5 R6
R5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms,
-(CH2)qC02R8, -(CH2)rNR9C02R10, carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazoylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R6 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms;
R7, R8, and R10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri- substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R11 and R12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R13 and R14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms; Y is CH or N; m is 0 - 4; n is 0 - 4; p is 1 - 2; q is 0 - 4; ris O - 4; t is O - 4; u is 0 - 4;
O II wherein R5, R6, m, and n are independent in each of the[C(CH2)mCH(CH2)nN]
I I
R5 R6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R1, R2> and R3 are not all hydrogen, further provided that R1, R2> and R3 are not all
O
II
- [C(CH2)mCH(CH2)-.N]pCO2R7 , and still further provided that t and u are not
I I
R5 R6 both 0 when X is O or S.
35. A pharmaceutical composition for the use in treating transplantation rejection, host vs. graft disease, autoimmune diseases, and diseases of inflammation in a mammal which comprises, an immunosuppressive amount of a compound having the structure
wherein R1, R2> and R3 are each, independently, hydrogen, or R4;
O
II O II
R4 is - [C(CH2)mCH(CH2)nN]pCO2R7 , -C-(CH2)tX(CH2)uCO2Rn , or
I I R5 R6
R5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms,
-(CH2)qC02R8, -(CH2)rNR9C02R10, carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazoylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R6 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms;
R7, R8, and R10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri- substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R11 and R12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R13 X is -C-, O, or S;
I R14
R13 and R14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms;
Y is CH or N; m is 0 - 4; n is 0 - 4; p is 1 - 2; q is 0 - 4; r is 0 - 4; tis 0 -4; u is 0 - 4;
O
II wherein R5, R6, m, and n are independent in each of the[C(CH2)mCH(CH2)nN]
I I
R5 R6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R1, R2- and R3 are not all hydrogen, further provided that R1, R2- and R3 are not all
O
II
— [C(CH2)mCH(CH2)nN]pCO2R7 , and still further provided that t and u are not
I I
R5 R6 both 0 when X is O or S.
36. A pharmaceutical composition for the use in treating fungal infections, which comprises an antifungal amount of a compound having the structure
wherein R1, R2- and R3 are each, independently, hydrogen, or R4;
O
II O
II
R4 is -[C(CH2)mCH(CH2)nN]pCO2R7 , -C- (CH2)tX(CH2)uCO2R11 or
I I
R5 R6
R5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms,
-(CH2)qC02R8, -(CH2)rNR9Cθ2R10, carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazoylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R6 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms; R7, R8, and R10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri- substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R11 and R12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R13 Xis -C-, O, or S;
I R14
R13 and R14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms; Y is CH orN; m is 0 - 4; n is 0 - 4; p is 1 - 2; q is 0 - 4; r is 0 - 4; t is O - 4; u is 0 - 4;
O
II wherein R5, R6, m, and n are independent in each of the[C(CH2)mCH(CH2)nN]
I I
R5 R6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R1, R2- and R3 are not all hydrogen, further provided that R1, R2> and R3 are not all
O
II
- [C(CH2)mCH(CH2)nN]pCO2R7 , and still further provided that t and u are not
I I
R5 R6 both 0 when X is O or S.
37. A process for preparing a compound of the formula
wherein R1, R2> and R3 are each, independently, hydrogen, or R4; O
II ?
R4 is - [C(CH2)mCH(CH2)nN]pCO2R7 , -C- (CH2)tX(CH2)uCO2Rn , or
I I R5 R6
R5 is hydrogen, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms,
-(CH2)qCO2R8, -(CH2)rNR9CO2R10, carbamylalkyl of 2-3 carbon atoms, aminoalkyl of 1-4 carbon atoms, hydroxyalkyl of 1-4 carbon atoms, guanylalkyl of 2-4 carbon atoms, mercaptoalkyl of 1-4 carbon atoms, alkylthioalkyl of 2-6 carbon atoms, indolylmethyl, hydroxyphenylmethyl, imidazoylmethyl or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R6 and R9 are each, independently, hydrogen, alkyl of 1-6 carbon atoms, or aralkyl of 7-10 carbon atoms;
R7, R8, and R10 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, fluorenylmethyl, or phenyl which is optionally mono-, di-, or tri- substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid; R11 and R12 are each, independently, alkyl of 1-6 carbon atoms, aralkyl of 7-10 carbon atoms, or phenyl which is optionally mono-, di-, or tri-substituted with a substituent selected from alkyl of 1-6 carbon atoms, alkoxy of 1-6 carbon atoms, hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 carbon atoms, trifluoromethyl, amino, or a carboxylic acid;
R13
X is -C-, O, or S;
I R14
R13 and R14 are each, independently, hydrogen or alkyl of 1-6 carbon atoms; Y is CH orN; m is 0 - 4; n is 0 - 4; p is 1 - 2; q is 0 - 4; r is 0 - 4; tis O - 4; u is 0 - 4;
O
II wherein R5, R6, m, and n are independent in each of the[C(CH2)mCH(CH2)nN]
I I
R5 R6 subunits when p = 2; or a pharmaceutically acceptable salt thereof, with the proviso that R1, R2- and R3 are not all hydrogen, further provided that R1, R2> and R3 are not all
O
II
- [C(CH2)mCH(CH2)nN]pC02R7 , and still further provided that t and u are not
I I
R5 R6 both 0 when X is O or S; which comprises (a) acylating rapamycin with an acylating agent or (b) sequentially acylating rapamycin with one or more acylating agents, said acylating agent(s) being selected from acids of formula: (CH2)tX(CH2)uCO2Rn , or
where Z is OH or reactive derivatives thereof, if desired protecting any of 42, 31 and 14 positions of rapamycin with an appropriate protecting group and removing said group as required, and further if desired isolating the product as a pharmaceutically acceptable salt
EP19910919248 1990-09-19 1991-09-19 Carboxylic acid esters of rapamycin Ceased EP0549727A1 (en)

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Families Citing this family (163)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0427680B1 (en) * 1989-11-09 1995-08-23 Sandoz Ltd. Heteroatoms-containing tricyclic compounds
NO921449L (en) * 1991-04-17 1992-10-19 American Home Prod CARBAMATES OF RAPAMYCIN
US5565560A (en) * 1991-05-13 1996-10-15 Merck & Co., Inc. O-Aryl,O-alkyl,O-alkenyl and O-alkynylmacrolides having immunosuppressive activity
US5262533A (en) * 1991-05-13 1993-11-16 Merck & Co., Inc. Amino O-aryl macrolides having immunosuppressive activity
US5247076A (en) * 1991-09-09 1993-09-21 Merck & Co., Inc. Imidazolidyl macrolides having immunosuppressive activity
US5284840A (en) * 1992-06-12 1994-02-08 Merck & Co., Inc. Alkylidene macrolides having immunosuppressive activity
US5284877A (en) * 1992-06-12 1994-02-08 Merck & Co., Inc. Alkyl and alkenyl macrolides having immunosuppressive activity
SG47580A1 (en) * 1992-10-13 1998-04-17 American Home Prod Carbamates of rapamycin
US5262423A (en) * 1992-10-29 1993-11-16 American Home Products Corporation Rapamycin arylcarbonyl and alkoxycarbonyl carbamates as immunosuppressive and antifungal agents
US5258389A (en) * 1992-11-09 1993-11-02 Merck & Co., Inc. O-aryl, O-alkyl, O-alkenyl and O-alkynylrapamycin derivatives
US5260300A (en) 1992-11-19 1993-11-09 American Home Products Corporation Rapamycin carbonate esters as immuno-suppressant agents
US5310901A (en) * 1993-03-05 1994-05-10 Merck & Co., Inc. O-heteroaryl, O-alkylheteroaryl, O-alkenylheteroaryl and O-alkynlheteroarylrapamycin derivatives
US5310903A (en) * 1993-03-05 1994-05-10 Merck & Co., Inc. Imidazolidyl rapamycin derivatives
GB9307491D0 (en) * 1993-04-08 1993-06-02 Sandoz Ltd Organic compounds
USRE40596E1 (en) * 1993-04-08 2008-12-02 Novartis Ag Rapamycin assay
US5504091A (en) * 1993-04-23 1996-04-02 American Home Products Corporation Biotin esters of rapamycin
ES2295093T3 (en) 1993-04-23 2008-04-16 Wyeth CONJUGATES AND RAPAMYCIN ANTIBODIES.
AU6711994A (en) * 1993-04-23 1994-11-21 American Home Products Corporation Rapamycin conjugates and antibodies
US7279561B1 (en) 1993-04-23 2007-10-09 Wyeth Anti-rapamycin monoclonal antibodies
US5527907A (en) * 1993-11-19 1996-06-18 Abbott Laboratories Macrolide immunomodulators
EP0729471A1 (en) * 1993-11-19 1996-09-04 Abbott Laboratories Semisynthetic analogs of rapamycin (macrolides) being immunomodulators
CN1046944C (en) * 1993-12-17 1999-12-01 山道士有限公司 Rapamycin derivatives useful as immunosuppressants
US5389639A (en) * 1993-12-29 1995-02-14 American Home Products Company Amino alkanoic esters of rapamycin
US5362718A (en) * 1994-04-18 1994-11-08 American Home Products Corporation Rapamycin hydroxyesters
US5693648A (en) * 1994-09-30 1997-12-02 Merck & Co., Inc. O-aryl, O-alkyl, O-alkenyl and O-alkynyl-macrolides having immunosuppressive activity
MY129435A (en) 1994-10-26 2007-04-30 Novartis Ag Pharmaceutical microemulsion preconcentrates
US6187757B1 (en) 1995-06-07 2001-02-13 Ariad Pharmaceuticals, Inc. Regulation of biological events using novel compounds
AU712193B2 (en) * 1995-06-09 1999-10-28 Novartis Ag Rapamycin derivatives
US5780462A (en) * 1995-12-27 1998-07-14 American Home Products Corporation Water soluble rapamycin esters
US7399480B2 (en) 1997-09-26 2008-07-15 Abbott Laboratories Methods of administering tetrazole-containing rapamycin analogs with other therapeutic substances using medical devices
US6015815A (en) * 1997-09-26 2000-01-18 Abbott Laboratories Tetrazole-containing rapamycin analogs with shortened half-lives
US6890546B2 (en) 1998-09-24 2005-05-10 Abbott Laboratories Medical devices containing rapamycin analogs
US20030129215A1 (en) 1998-09-24 2003-07-10 T-Ram, Inc. Medical devices containing rapamycin analogs
US7357942B2 (en) 1997-09-26 2008-04-15 Abbott Laboratories Compositions, systems, and kits for administering zotarolimus and paclitaxel to blood vessel lumens
US8257725B2 (en) 1997-09-26 2012-09-04 Abbott Laboratories Delivery of highly lipophilic agents via medical devices
US8394398B2 (en) 1997-09-26 2013-03-12 Abbott Laboratories Methods of administering rapamycin analogs with anti-inflammatories using medical devices
US8057816B2 (en) 1997-09-26 2011-11-15 Abbott Laboratories Compositions and methods of administering paclitaxel with other drugs using medical devices
US8257726B2 (en) 1997-09-26 2012-09-04 Abbott Laboratories Compositions, systems, kits, and methods of administering rapamycin analogs with paclitaxel using medical devices
US7378105B2 (en) 1997-09-26 2008-05-27 Abbott Laboratories Drug delivery systems, kits, and methods for administering zotarolimus and paclitaxel to blood vessel lumens
US8257724B2 (en) 1998-09-24 2012-09-04 Abbott Laboratories Delivery of highly lipophilic agents via medical devices
US7960405B2 (en) 1998-09-24 2011-06-14 Abbott Laboratories Compounds and methods for treatment and prevention of diseases
US7455853B2 (en) 1998-09-24 2008-11-25 Abbott Cardiovascular Systems Inc. Medical devices containing rapamycin analogs
US8124630B2 (en) 1999-01-13 2012-02-28 Bayer Healthcare Llc ω-carboxyaryl substituted diphenyl ureas as raf kinase inhibitors
EP2298311B1 (en) 1999-01-13 2012-05-09 Bayer HealthCare LLC w-Carboxy aryl substituted diphenyl ureas as p38 kinase inhibitors
AU6643200A (en) * 1999-08-18 2001-03-13 American Home Products Corporation Water soluble sdz-rad esters
DE60010098T2 (en) 1999-08-24 2005-03-31 Ariad Gene Therapeutics, Inc., Cambridge 28-EPIRAPALOGE
US7067526B1 (en) 1999-08-24 2006-06-27 Ariad Gene Therapeutics, Inc. 28-epirapalogs
GB0008785D0 (en) 2000-04-10 2000-05-31 Novartis Ag Organic compounds
ES2313983T3 (en) * 2000-09-19 2009-03-16 Wyeth RAPAMYCIN HYDROSOLUBLE ESTERS.
MXPA04007832A (en) 2002-02-11 2005-09-08 Bayer Pharmaceuticals Corp Aryl ureas with angiogenesis inhibiting activity.
CN100349627C (en) 2002-09-06 2007-11-21 艾博特公司 Medical devices containing rapamycin analogs
AU2003293529A1 (en) 2002-12-16 2004-07-29 Nitromed, Inc. Nitrosated and nitrosylated rapamycin compounds, compositions and methods of use
JP2006516548A (en) 2002-12-30 2006-07-06 アンジオテック インターナショナル アクツィエン ゲゼルシャフト Drug delivery from rapidly gelled polymer compositions
US7160867B2 (en) 2003-05-16 2007-01-09 Isotechnika, Inc. Rapamycin carbohydrate derivatives
CA2526617C (en) 2003-05-20 2015-04-28 Bayer Pharmaceuticals Corporation Diaryl ureas with kinase inhibiting activity
TW200517114A (en) 2003-10-15 2005-06-01 Combinatorx Inc Methods and reagents for the treatment of immunoinflammatory disorders
US20060211752A1 (en) 2004-03-16 2006-09-21 Kohn Leonard D Use of phenylmethimazoles, methimazole derivatives, and tautomeric cyclic thiones for the treatment of autoimmune/inflammatory diseases associated with toll-like receptor overexpression
US8431145B2 (en) 2004-03-19 2013-04-30 Abbott Laboratories Multiple drug delivery from a balloon and a prosthesis
EP1737869A1 (en) * 2004-04-14 2007-01-03 Wyeth a Corporation of the State of Delaware Regiospecific synthesis of rapamycin 42-ester derivatives
WO2005105812A1 (en) 2004-04-14 2005-11-10 Wyeth Process for preparing rapamycin 42-esters and fk-506 32-esters with dicarboxylic acid, precursors for rapamycin conjugates and antibodies
EP1768662A2 (en) 2004-06-24 2007-04-04 Novartis Vaccines and Diagnostics, Inc. Small molecule immunopotentiators and assays for their detection
KR101387456B1 (en) 2005-02-09 2014-04-21 산텐 세이야꾸 가부시키가이샤 Liquid formulations for treatment of diseases or conditions
JP2008533007A (en) 2005-03-07 2008-08-21 ワイス Oxepane isomer of 42-O- (2-hydroxy) ethyl-rapamycin
JP5242374B2 (en) 2005-03-23 2013-07-24 アボット・ラボラトリーズ Composition and method of administering rapamycin analogs using medical devices for long-term efficacy
EP1868663B1 (en) 2005-03-23 2011-11-16 Abbott Laboratories Delivery of highly lipophilic agents via medical devices
KR20130087058A (en) 2005-07-20 2013-08-05 노파르티스 아게 Combination of a pyrimidylaminobenzamide and an mtor kinase inhibitor
JP2009514969A (en) 2005-11-09 2009-04-09 コンビナトアールエックス インコーポレーティッド Methods, compositions, and kits for treating medical conditions
EP1954685A4 (en) 2005-11-16 2009-11-11 Nitromed Inc Furoxan compounds, compositions and methods of use
CA2629245C (en) 2005-11-21 2016-07-12 Novartis Ag Neuroendocrine tumor treatment
US7700614B2 (en) 2005-12-14 2010-04-20 Abbott Laboratories One pot synthesis of tetrazole derivatives of rapamycin
EP2001438A2 (en) 2006-02-09 2008-12-17 Macusight, Inc. Stable formulations, and methods of their preparation and use
US7678901B2 (en) 2006-02-28 2010-03-16 Wyeth Rapamycin analogs containing an antioxidant moiety
US7622477B2 (en) 2006-02-28 2009-11-24 Cordis Corporation Isomers and 42-epimers of rapamycin alkyl ether analogs, methods of making and using the same
US20070203169A1 (en) * 2006-02-28 2007-08-30 Zhao Jonathon Z Isomers and 42-epimers of rapamycin ester analogs, methods of making and using the same
DK2001466T3 (en) 2006-03-23 2016-02-29 Santen Pharmaceutical Co Ltd LOW-DOSAGE RAPAMYCINE FOR TREATMENT OF VASCULAR PERMEABILITY-RELATED DISEASES
US7883855B2 (en) 2006-07-21 2011-02-08 Abbott Laboratories Immunosuppressant drug extraction reagent for immunoassays
WO2008042216A2 (en) 2006-09-28 2008-04-10 Follica, Inc. Methods, kits, and compositions for generating new hair follicles and growing hair
US10265407B2 (en) 2007-02-15 2019-04-23 Yale University Modular nanodevices for smart adaptable vaccines
EP2232264B1 (en) 2007-12-19 2015-12-02 Abbott Laboratories Immunosuppressant drug extraction reagent for immunoassays
EP2245165A4 (en) 2008-01-11 2011-06-01 Massachusetts Eye & Ear Infirm Conditional-stop dimerizable caspase transgenic animals
US20100048912A1 (en) 2008-03-14 2010-02-25 Angela Brodie Novel C-17-Heteroaryl Steroidal CYP17 Inhibitors/Antiandrogens, In Vitro Biological Activities, Pharmacokinetics and Antitumor Activity
US20110312916A1 (en) 2009-02-05 2011-12-22 Tokai Pharmaceuticals, Inc. Novel prodrugs of steroidal cyp17 inhibitors/antiandrogens
US10485792B2 (en) 2009-08-26 2019-11-26 Hadasit Medical Research Services & Development Limited Sustained release delivery systems for the prevention and treatment of head and neck cancers
US8951595B2 (en) 2009-12-11 2015-02-10 Abbott Cardiovascular Systems Inc. Coatings with tunable molecular architecture for drug-coated balloon
JP5918214B2 (en) 2010-04-13 2016-05-18 ノバルティス アーゲー Combinations comprising a cyclin dependent kinase 4 or cyclin dependent kinase (CDK4 / 6) inhibitor and an MTOR inhibitor for treating cancer
NZ602807A (en) 2010-04-16 2014-02-28 Novartis Ag 4-amino-5-fluoro-3-[6-(4-methylpiperazin-1-yl)-1h-benzimidazol-2-yl]-1h-quinolin-2-one and an mtor inhibitor
KR101253399B1 (en) 2010-10-26 2013-04-11 영남대학교 산학협력단 Rapamycin formulation encapsulated in reconstituted high-density lipoprotein containing V156K-apoA-I
PT2683406T (en) 2011-03-11 2019-07-08 Beth Israel Deaconess Medical Ct Inc Anti-cd40 antibodies and uses thereof
JP6086902B2 (en) 2011-04-25 2017-03-01 ノバルティス アーゲー Combination of phosphatidylinositol-3-kinase (PI3K) inhibitor and mTOR inhibitor
CN104519907A (en) 2012-03-23 2015-04-15 昆士兰大学 Immunomodulatory agent and uses therefor
US9597385B2 (en) 2012-04-23 2017-03-21 Allertein Therapeutics, Llc Nanoparticles for treatment of allergy
WO2013192367A1 (en) 2012-06-22 2013-12-27 Novartis Ag Neuroendocrine tumor treatment
EP2919759A4 (en) 2012-11-14 2016-07-20 Ohio State Innovation Foundation Materials and methods useful for treating glioblastoma
SG11201507093WA (en) 2013-03-14 2015-10-29 Univ Maryland Baltimore Office Of Technology Transfer Androgen receptor down-regulating agents and uses thereof
AU2014248090B2 (en) 2013-04-03 2018-08-02 N-Fold Llc Novel nanoparticle compositions
WO2014184734A1 (en) 2013-05-14 2014-11-20 Novartis Ag Markers associated with mtor inhibition
US9593356B2 (en) 2013-06-11 2017-03-14 Takara Bio Usa, Inc. Protein enriched microvesicles and methods of making and using the same
RU2016104643A (en) 2013-08-12 2017-09-19 Токай Фармасьютикалз, Инк. BIOMARKERS FOR THE TREATMENT OF NEOPLASTIC DISEASES WITH THE APPLICATION OF METHODS TREATED AT ANDROGEN
WO2015044854A1 (en) 2013-09-24 2015-04-02 Novartis Ag Markers associated with mtor inhibition
EP3104890A1 (en) 2014-02-11 2016-12-21 Novartis AG Pharmaceutical combinations comprising a pi3k inhibitor for the treatment of cancer
AU2015237200A1 (en) 2014-03-27 2016-10-06 The Brigham And Women's Hospital, Inc. Metabolically-activated drug conjugates to overcome resistance in cancer therapy
WO2015171723A1 (en) 2014-05-06 2015-11-12 Research Development Foundation Methods for treating insulin resistance and for sensitizing patients to glp1 agonist therapy
CN105461738B (en) * 2014-06-03 2019-03-08 中国人民解放军军事医学科学院毒物药物研究所 A kind of rapamycin derivative, preparation method, its pharmaceutical composition and purposes
CN113620978A (en) 2014-09-11 2021-11-09 加利福尼亚大学董事会 mTORC1 inhibitors
CN116063481A (en) 2015-09-04 2023-05-05 普里玛托普医疗股份有限公司 Humanized anti-CD 40 antibodies and uses thereof
US20190290631A1 (en) 2016-05-27 2019-09-26 Nippon Kayaku Kabushiki Kaisha Pharmaceutical composition comprising rapamycin or derivative thereof
US10682340B2 (en) 2016-06-30 2020-06-16 Durect Corporation Depot formulations
CA2972296A1 (en) 2016-06-30 2017-12-30 Durect Corporation Depot formulations
RS62456B1 (en) 2016-12-22 2021-11-30 Amgen Inc Benzisothiazole, isothiazolo[3,4-b]pyridine, quinazoline, phthalazine, pyrido[2,3-d]pyridazine and pyrido[2,3-d]pyrimidine derivatives as kras g12c inhibitors for treating lung, pancreatic or colorectal cancer
EA201990127A1 (en) 2016-12-30 2020-08-18 Дьюрект Корпорейшн DEPO-PREPARATION
JOP20190272A1 (en) 2017-05-22 2019-11-21 Amgen Inc Kras g12c inhibitors and methods of using the same
SG11202001499WA (en) 2017-09-08 2020-03-30 Amgen Inc Inhibitors of kras g12c and methods of using the same
AU2019262979B2 (en) 2018-05-01 2023-07-06 Revolution Medicines, Inc. C26-linked rapamycin analogs as mTOR inhibitors
US20190336609A1 (en) 2018-05-01 2019-11-07 Revolution Medicines, Inc. C40-, C28-, and C-32-Linked Rapamycin Analogs as mTOR Inhibitors
EP3788038B1 (en) 2018-05-04 2023-10-11 Amgen Inc. Kras g12c inhibitors and methods of using the same
US11045484B2 (en) 2018-05-04 2021-06-29 Amgen Inc. KRAS G12C inhibitors and methods of using the same
WO2019217691A1 (en) 2018-05-10 2019-11-14 Amgen Inc. Kras g12c inhibitors for the treatment of cancer
US11096939B2 (en) 2018-06-01 2021-08-24 Amgen Inc. KRAS G12C inhibitors and methods of using the same
EP3802537A1 (en) 2018-06-11 2021-04-14 Amgen Inc. Kras g12c inhibitors for treating cancer
MX2020012261A (en) 2018-06-12 2021-03-31 Amgen Inc Kras g12c inhibitors encompassing a piperazine ring and use thereof in the treatment of cancer.
JP2020090482A (en) 2018-11-16 2020-06-11 アムジエン・インコーポレーテツド Improved synthesis of key intermediate of kras g12c inhibitor compound
CA3117222A1 (en) 2018-11-19 2020-05-28 Amgen Inc. Kras g12c inhibitors and methods of using the same
JP7377679B2 (en) 2018-11-19 2023-11-10 アムジエン・インコーポレーテツド Combination therapy comprising a KRASG12C inhibitor and one or more additional pharmaceutically active agents for the treatment of cancer
MA54546A (en) 2018-12-20 2022-03-30 Amgen Inc HETEROARYL AMIDES USEFUL AS KIF18A INHIBITORS
PE20211475A1 (en) 2018-12-20 2021-08-05 Amgen Inc KIF18A INHIBITORS
ES2953821T3 (en) 2018-12-20 2023-11-16 Amgen Inc KIF18A inhibitors
MA54547A (en) 2018-12-20 2022-03-30 Amgen Inc HETEROARYL AMIDES USEFUL AS KIF18A INHIBITORS
JP2022522777A (en) 2019-03-01 2022-04-20 レボリューション メディシンズ インコーポレイテッド Bicyclic heteroaryl compounds and their use
US20230096028A1 (en) 2019-03-01 2023-03-30 Revolution Medicines, Inc. Bicyclic heterocyclyl compounds and uses thereof
EP3738593A1 (en) 2019-05-14 2020-11-18 Amgen, Inc Dosing of kras inhibitor for treatment of cancers
EP3972973A1 (en) 2019-05-21 2022-03-30 Amgen Inc. Solid state forms
US20220372018A1 (en) 2019-08-02 2022-11-24 Amgen Inc. Kif18a inhibitors
WO2021026098A1 (en) 2019-08-02 2021-02-11 Amgen Inc. Kif18a inhibitors
AU2020324963A1 (en) 2019-08-02 2022-02-24 Amgen Inc. KIF18A inhibitors
CN114401953A (en) 2019-08-02 2022-04-26 美国安进公司 KIF18A inhibitors
WO2021081212A1 (en) 2019-10-24 2021-04-29 Amgen Inc. Pyridopyrimidine derivatives useful as kras g12c and kras g12d inhibitors in the treatment of cancer
IL292438A (en) 2019-10-28 2022-06-01 Merck Sharp & Dohme Small molecule inhibitors of kras g12c mutant
US20230023023A1 (en) 2019-10-31 2023-01-26 Taiho Pharmaceutical Co., Ltd. 4-aminobut-2-enamide derivatives and salts thereof
CA3159559A1 (en) 2019-11-04 2021-05-14 Revolution Medicines, Inc. Ras inhibitors
EP4054719A1 (en) 2019-11-04 2022-09-14 Revolution Medicines, Inc. Ras inhibitors
TW202132314A (en) 2019-11-04 2021-09-01 美商銳新醫藥公司 Ras inhibitors
PE20230249A1 (en) 2019-11-08 2023-02-07 Revolution Medicines Inc BICYCLIC HETEROARYL COMPOUNDS AND THEIR USES
WO2021097212A1 (en) 2019-11-14 2021-05-20 Amgen Inc. Improved synthesis of kras g12c inhibitor compound
AR120456A1 (en) 2019-11-14 2022-02-16 Amgen Inc ENHANCED SYNTHESIS OF KRAS G12C INHIBITOR COMPOUND
JP2023505100A (en) 2019-11-27 2023-02-08 レボリューション メディシンズ インコーポレイテッド Covalent RAS inhibitors and uses thereof
WO2021106231A1 (en) 2019-11-29 2021-06-03 Taiho Pharmaceutical Co., Ltd. A compound having inhibitory activity against kras g12d mutation
BR112022010086A2 (en) 2020-01-07 2022-09-06 Revolution Medicines Inc SHP2 INHIBITOR DOSAGE AND CANCER TREATMENT METHODS
US20230174518A1 (en) 2020-04-24 2023-06-08 Taiho Pharmaceutical Co., Ltd. Kras g12d protein inhibitors
US20230181536A1 (en) 2020-04-24 2023-06-15 Taiho Pharmaceutical Co., Ltd. Anticancer combination therapy with n-(1-acryloyl-azetidin-3-yl)-2-((1h-indazol-3-yl)amino)methyl)-1h-imidazole-5-carboxamide inhibitor of kras-g12c
AU2021308045A1 (en) 2020-07-15 2023-02-23 Taiho Pharmaceutical Co., Ltd. Pyrimidine compound-containing combination to be used in tumor treatment
AU2021344830A1 (en) 2020-09-03 2023-04-06 Revolution Medicines, Inc. Use of SOS1 inhibitors to treat malignancies with SHP2 mutations
KR20230067635A (en) 2020-09-15 2023-05-16 레볼루션 메디슨즈, 인크. Indole derivatives as RAS inhibitors in the treatment of cancer
CN117396472A (en) 2020-12-22 2024-01-12 上海齐鲁锐格医药研发有限公司 SOS1 inhibitors and uses thereof
CN117500811A (en) 2021-05-05 2024-02-02 锐新医药公司 Covalent RAS inhibitors and uses thereof
CR20230570A (en) 2021-05-05 2024-01-22 Revolution Medicines Inc Ras inhibitors
WO2022235870A1 (en) 2021-05-05 2022-11-10 Revolution Medicines, Inc. Ras inhibitors for the treatment of cancer
EP4347041A1 (en) 2021-05-28 2024-04-10 Taiho Pharmaceutical Co., Ltd. Small molecule inhibitors of kras mutated proteins
AR127308A1 (en) 2021-10-08 2024-01-10 Revolution Medicines Inc RAS INHIBITORS
TW202340214A (en) 2021-12-17 2023-10-16 美商健臻公司 Pyrazolopyrazine compounds as shp2 inhibitors
EP4227307A1 (en) 2022-02-11 2023-08-16 Genzyme Corporation Pyrazolopyrazine compounds as shp2 inhibitors
WO2023172940A1 (en) 2022-03-08 2023-09-14 Revolution Medicines, Inc. Methods for treating immune refractory lung cancer
WO2023240263A1 (en) 2022-06-10 2023-12-14 Revolution Medicines, Inc. Macrocyclic ras inhibitors
WO2024081916A1 (en) 2022-10-14 2024-04-18 Black Diamond Therapeutics, Inc. Methods of treating cancers using isoquinoline or 6-aza-quinoline derivatives

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4316885A (en) * 1980-08-25 1982-02-23 Ayerst, Mckenna And Harrison, Inc. Acyl derivatives of rapamycin
US4650803A (en) * 1985-12-06 1987-03-17 University Of Kansas Prodrugs of rapamycin
JPH04230389A (en) * 1990-07-16 1992-08-19 American Home Prod Corp Rapamycin derivative
JP3140228B2 (en) * 1992-02-17 2001-03-05 ファイザー製薬株式会社 Novel macrocyclic lactone and its producing bacteria

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9205179A1 *

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