CA2266039A1 - Alkylated rapamycin derivatives - Google Patents
Alkylated rapamycin derivatives Download PDFInfo
- Publication number
- CA2266039A1 CA2266039A1 CA002266039A CA2266039A CA2266039A1 CA 2266039 A1 CA2266039 A1 CA 2266039A1 CA 002266039 A CA002266039 A CA 002266039A CA 2266039 A CA2266039 A CA 2266039A CA 2266039 A1 CA2266039 A1 CA 2266039A1
- Authority
- CA
- Canada
- Prior art keywords
- alkyl
- ch2ch2o
- independently
- ch2ch2ch2o
- phenyl
- 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.)
- Abandoned
Links
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
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/06—Antiasthmatics
-
- 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
- A61P31/10—Antimycotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
Landscapes
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Pulmonology (AREA)
- Transplantation (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
Abstract
This invention relates to compounds which possess immunosuppressive and/or anti tumor and/or antiinflammatory activity in vivo and/or inhibit thymocyte proliferation in vitro. These compounds are therefore useful in the treatment of transplantation rejection, autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, multiple sclerosis and in the treatment of Candida albicans infections and also in treatment of diseases of inflammation.
These compounds are represented by formula (I), wherein W and Y are OR1 and X
and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein: R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)nCH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene; -CH2(CH2)n-OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl; and -CH2(CH2)n-X where X is F, Cl, Br or I; R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR3, -CH2(CH2)n-X where X
is F, Cl, Br or I; and -(CH2)nCH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar; n = 1-10 independently; m = 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl; 1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
These compounds are represented by formula (I), wherein W and Y are OR1 and X
and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein: R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)nCH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene; -CH2(CH2)n-OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl; and -CH2(CH2)n-X where X is F, Cl, Br or I; R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR3, -CH2(CH2)n-X where X
is F, Cl, Br or I; and -(CH2)nCH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar; n = 1-10 independently; m = 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl; 1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
Description
W 0~ 337~ PCTAJS97/15439 ALKYLATE:D RAPAMYCIN DERIVATIVES
This invention relates to compounds of formula I below or pharm~eutir~l1y acceptable salts thereof which possess immunosuppressive and/or anti tumor and/or s antiinflamrnatory activity in vivo and/or inhibit thymocyte proliferation in vilro. These compounds are therefore useful in the treatment of transplantation rejection, autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, multiple sclerosis, the treatn1ellt of Candicia ~1lbicans infectiolls, treatment of diseases of inflammation neatment of hyperproliferative vascular disease (restenosis) and in the treatrnent of certain human tumors.
BACKGROUND OF THE INVENTION
Rapamycin is a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus, which was follnd to have antifungal activity, particularly against Candida albicans, both in vitro alld in vlvo ~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 (197~); U.S. Patent 3,929,992; and U.S. Patent 3,993,749].
,~- OCH3 4% ~37 26 ,48 8 [~ 2~OH
~0 ~ CH30- ~
HO ~J~4 - 34 46--~ 013 OCH3 5(~ ~ 35 ~ 1 4 2 4; 51 20Rap~mycin (Positions numbered accordillg to Chemical Abstract~s) Rapamycin alone (U.S. Patent 4,885,171) or in combination with picibanil (U.S. Patent 4,401,653) has been shown to have antitumor activity. R. Martel et al.
I
wo g8,~597., rCT/US97/15439 [Can. J. Physiol. Ph~rmacol. 55, 48 (1977)1 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.
s The immunosuppressive effects of rapamycin have been disclosed in FASEB 3, 3411 (19~9). Rapamycin has been shown to be effective in inhibiting transplant rejection (U.S. Patent 5,10(),899). Cyclosporin A and FK-506, other macrocyclic molecules, also have been shown to be effective as immunosuppressive agents, therefore useful in preventing transplant rejection IFASEB 3, 3411 (1989); FASEB 3, o 5256 (1989); and R. Y. Calne et al., Lancet 1183 (1978)]. U. S. patent 5,321,009 discloses a method of prophylactica]ly preventing the onset, preventing the development, and arresting the progression of insulin-dependent diabetes mellitus by administration of rapamycin. U. S. patellt 5,2~,711 discloses a method of preventing or treating hyperproliferative vascular disease by administration of a combination of s rapamycin and heparin. U. S. patent 5,2~6,730 discloses a method of treatingimmunoinflammatory disease by treatlllent with rapamycin alone or in combination with cyclosporin A. U. S. patent 5,2~6,731 provides a method of treating immunoinflammatory bowel disease by administration of rapamycin alone or in combination with cyclosporin A.
Various structural features of rapamycin have been modified in efforts to increase the potency or specificity of phamlacological action. For instance, a number of U. S. patents disclose compounds where one or more of the hydroxy groups having normal stereochemistry at positions 14, 31, and 42 have been converted into acylesters, sulfonyl esters, and carballlates. U. S. patent 5,023,263 discloses 42-oxo 2s rapamycin. V. S. patent 5,258,389 discloses 31 and/or 42 O-alkyl, O-aryl, O-alkenyl, and O-alkynyl ethers of rapamycin having normal stereochemistry at the 42 position.
The PCI' published applicatioll WO 94/~)9()1() discloses 31 and/or 42 O-alkylated rapamycin analogs wherein the keto groups at positions 15 and 33 may be reduced to a hydroxyl group or a methylene group.
SUMMARY OF THE INVENTION
The compounds of this invention are represented by the following chemical formula and are novel. While a number of rapamycin analogs substituted at positions 3s 31 and 42 have been disclosed, substitution at position 29 has not been heretofore disclosed.
W O ~8J~/V PCT~US97/15439 The compounds useful in this invention are represented by the formula below:
~3 W
38 ' ~ - OCH3 3 ~ ~ z~ Z y ~~o ~ CH303 33~
HO~I~4 134 46 ~ 013 OCH3 5( ~ > 35 11 ~ ' ~ 36 4~ 51 wherein W and Y are ORl and X and Z together form a bond or W and X are oR2 and Y and Z together fonn ,t bond, wherein R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O~nCH3 where n is not 1, I O -CH2CH2CH20(CH2CH20)m-CH3, ~(CH2)n-CH2CH(oR3)cH2oR4 where R3 ;~nd R4 are H, Cl-Clo alkyl, or R3 and R4 together ale ethylene, metl~ylene or dimethylmethylene;
~CH2(CH2)n-oR3 where R3 is not H, Cl-Cl() alkyl, or C(O)C1-CIo alkyl, and -CH2(CH2)n-X where X is F, Cl, Br or l;
R2 is selected from H, Cl-Clo alkyl, Ar(CH2)n-, C3-CIo alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH20(CH2CH20)m-CH3, ~CH2(CH2)n-oR3~ -CH2(CH2)n-X
where X is F, Cl, Br or ~;
and -(CH2)n-CH~CH(ORS)CH2OR6 where RS and R6 are selected independently from H, Cl-Cl() alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -CoR7 and -Co2R7~ where R7 is Cl-C6 alkyl, C2-C6 alkellyl, or Ar;
n= 1-10 indepelldelltly: and 2s . _ W O 9~1~3~,~ PCTrUS97/15439 m= 1-5 independently;
In the above definitions of variable substituents, the term alkyl includes straight and branched chain hydrocarbons and the aryl groups (Ar) are selected independently s from phenyl, pyridinyE, quinolinyl, indolyl, furanyl, 1, 2, 3-triazolyl and tetrazolyl.
This invention also encompasses the pharmaceutica]ly acceptable acid addition salts where ehey can be formed.
The compounds of this invention exhibit immunosuppressive and/or antifungal and/or antitumor and/or antiinflammatory activity in vivo and/or inhibit thymocyte o proliferation in vitro and are therefore useful in the ùeatment or inhibition of organ or tissue transplantatioll rejection or host vs. graft disease, proliferative diseases such as restenosis following angioplasty procedures, autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, and multiple sclerosis; fungal infections, and diseases of inflamm.ltioll such as psoriasis, exzema, sebolThea, inflammatory bowel disease and pulmonary inflammation such ;lS asthma, chronic obstructive pulmonary disease, emphysema, brollchitis and the like. An invention compound was found toinhibit the growth of human breast, colon and ovarian cancer cell lines in submicromolar concentration and it is therefole expected that the invention compounds will be useful in inhibiting these and other tumors in humans.
Detailed Description of the Invention The compounds of this invention can be prepared by standard literature procedures as outlined below. Other suitable bases may be used in place of the 2,6-di-tert-butyl-4-methylpyridine and may include pyridine, lutidine, co]lidine, sodium hydride, or sodiulll carbonate.
f ~0 H ~O R
~ OMe ~~ OMe ~, '2 6 di(t buRty~l) 4-2rnetnylpyridine CH2cl2 ~ I n ¦l SSlr~ ~ ~;0 or ROC(=NH)CCI3, catalytic SSlr ~31 M e O J~ CF3SO3H, benzene o M e O' ~ ~
+ 31-substituted product and 31,42-disubstituted product W O 98/09970 PCTAUS97/lS439 Also, unexpectedly, a product was i.solated (1,3-allyl rearrangement) via triflation at C-28 followed by attack by external nucleophile.
OH ~OH
'J~ OMe ~ OMe , ~2,6-di(t-butyl)-4-methylpyridine ~" ~O R
n 1l CH2CI2 I n ~ 1 SSlr ~31or ROC(=NH)CCI3, catalytic Ss~r ~
M e O ~CF3SO3H, benzene o M e O~ J~~
~ss ~ss The compounds of this invention exhibit immunosuppressive and/or antifungal and/or antitumor and/or antiinflammatory activity in vivo andJor inhibit thymocyte proliferation in vitro and are therefore useful in the treatment of transplantation rejection, autoimmul1e dise~ces (i.e. Iupus, rheumatoid arthritis, diabetes mellitus, multiple sclerosis), Candida al~ica~Ls infections, and diseases of inflammation. An invention compound inhibits in vitro in sub-micron-olar concentrations, the growth of certain human tumor cell lines, including colon (MIP 101), breast (T47D, SKBR-3), and ovarian (A2780S) cells and therefore it is expected that compounds of this inventon will be useful in the treatment of these and other tumors.
The following procedllres are included to exemplify the preparation of inventioncompounds and employ stand.ud labor.ltory techniq~les known to those skilled in the art of organic systhesis.
Example ].
29-O-Ben~,ylI apamycin Toasolution of triflic anhydride (0.19 n-L, 1.13 mmol) in dichloromethane (5.0 rnL) in a round bottom flask (50 mL, flame dried) equipped with a magnetic stirrer at 0 ~C
was added 2,6-di-t-butyl-4-methyl pyridine (().253 g, 1.23 mmol) portionwise. The 2s reaction mixture was deg.lssed, and purged with nitrogen. To the solution was added a solution of benzyl alcohol (().1] mL, 1.06 mmol, diluted in 3 mL dichloromethane) dropwise. The solutioll became a pale white suspensioIl. The reaction was stirred at 0 ~C for 30 min. TLC analysis indicated no residllal benzyl alcohol remained. To the suspension was added another portion of 2,6-di-t-butyl-4-methyl pyridine (0.335 g, 1.63 mmol) portionwise, followed by a solution of rapamycin (0.855 g, 0.94 mmol, in 3 mL dichloromethalle) dropwise. The reaction was stirred at 0 ~C for 30 min, and warmed up to room temperature and stirred overnight. The reaction was quenched with saturated aqueous NaHC03, and the organic and aqueous layers were separated. Iheaqueous layer was extracted three times with ethyl acetate. The organic layers were combined, washed with brine, and dried over sodium sulfate. The solution was filtered and concentrated ln vac~o to afford a pale yellow foam. The product rnixture wasseparated by HPLC (40 % EtOAc/hexane, Dynan1ax 2" silica colun1n, 20 mL/min), and o four fractions were collected. The second fraction which was obtained in 12 % yield was identified as 42-O-benzylrapamycin. The third fraction which was obtained in 14 % yield was identified as the dihydr;lte of the title compound.
H NMR (4()() MHz, DMSO-d6) ~ 7.3~ (m, 5 H, PhCH2O-), 6.]() - 6.42 (m, 4 H, S vinylic), 5.48 (q, 1 H, vinylic), 4.95 (m, 2 H, PhCH20-), 3.32 (s, 3 H, -OC~13), 3.18 (s, 3 H, -OC~3), 3.()5 (s, 3 H, -OCH3); IR (KBr, cm~l): 3420, 2920, 1730, 1645, 1445; MS (neg. FAB): 1()03.4 lMl-, 652, 59(), 349; Anal. Calcd. for Cs3H83Nol3. 2 H2O: C 66.98 %, H 8.56 %, N 1.39 %; Found: C 66.53 %, H 8.51 %, N 1.70 %.
Preparation of substituted alkyl triflates.
To a solution of substituted alkanol (l.()() mmol) in dichloromethane (15.0 mL) in a round bottom flask (25 mL, flame dried) equipped with a magnetic stirrer at roomtemperature was added 2,~-di-t-butyl-4-metllyl pyridine (0.~50 g, 2.92 mmol). The 2s reaction mixture was degassed, purged with nitrogen, and cooled to -78 ~C. To the solution was added dropwise trifluoromethanes-llfonic anhydride (0.170 mL, 0.282 g, 1-.00 mmol) over a period of 5 minutes. The solution became a suspension. The reaction was stirred at () ~C for 3() minutes, then wamled up to room t~lllpe.dlul~. TLC
analysis showed completion of ttle reaction.
General Procedure for the Preparatioll of 42-substituted Derivatives of Rapamycin via Nucleophilic Substitution.
To the solution of substituted alkyl-triflate prep~ared from 1 mmol of alkanol was added at room temperature (unless otherwise noted) rap~mycin (914 mg, 1 mmol). The mixture was stirred between 4 and 12() h (monitoring of the extent of the reaction was ~, wo ~ 3~7~ PCT/USg7/I5439 carried out by TLC). When desired convelsioll has been achieved the reaction wasquenched with saturated aqueous NaHCO3, and the organic and aqueous layers were separated. The aqueous layer was extracted three times with ethyl acetate. The organic layers were combined, washed with brine, and dried over sodium sulfate. The solution s was filtered and concen~ated i~ vacuo to afford a product. Pure products were isolated by HPLC (norrnal phase- Dynamax 2" silica column, eluent EtOAc: hexane,20 mL4min; reversed ph~se -Dynamax 2" C18 column, eluent MeCN: water, 20 mL/min. Spectroscopic analyses were used to confirm the structures.
0 Example 2.
42-0-(4-chlorobutyl )rapamycin Method A. Alkanol used: 1()8 mg (I mmol) of l-chloro-butane-4-ol. Reaction tirne:
S 72 h at 25 ~C. Separatioll tech~ u~ en1ployed: 2" Dynal~ x silica column, eluent 40%
EtOAc: hexane,20 mL/min. Yield of product: 18() mg (18 %).
Spectral data follows: IH NMR (400 MHz, DMSO-d6 ~ 6.08-6.45 (m, 4 H), 3.4-3.7 (m, 8 H), 3.1S (s, 3H), 3.04 (s, 3 H). IR (KBr, cm~l) 3420, 2930, 1715, 1650, 1620, 1460. MS (neg. FAB) 1003.4 ~MI-, 590.2, 411.2. Anal. calcd. for CssHg6NO13CI: C 65.75 %, H 8.63 %, N 1.39 %; Found: C 65.14 %, H 8.64 %, N 1.18 %.
Example 3.
2s 42-0-(2-[2-(2-methoxy etho~y)-ethoxy]-ethyl)-l apamycin Method A. Alkanol used: 165 mg (I mmol) of triethylenegycol monomethyl ether.
Reaction time: 12() h ~I 1() ~C. Separation techni~lue employed: 2" Dynamax silica column, eluent 7()~c EtOAc: hexane,2() mL/min. Yield of product: 210 mg (19 %).
Spectral data follows: ~H NMR (4()() MHz, DMSO-d6 ~ 6.()5-6.45 (m, 4 H), 3.4-3.65 (m, 12 H), 3.23 (s, 3 H), 3.14 (s, 3 H), 3.()4 (s, 3 H). IR (KBr, cm~l ) 3420, 2925, 1720, 1645, 145(); MS (neg. FAB) 1()59.4 IM]-, 590.2, 467.2. Anal. Calcd.
3s for C5XHg3NOl6. H~O C 64.6() %, H 8.81 %, N 1.28 %; Found: C 63.96 %, H
8.64 %, N 1.15 %.
W O 9~ 7v PCT~US97/15439 Pharmacolo~y Immunosuppressive activity was evalu~ted in an in vitro standard s pharmacological test procedllre to measure Iymphocyte proliferation (LAF), in an in vivo procedure to evaluate the survival time of a pinch skin graft, and in an in vivo procedure to determine inhibition of T-cell mediated inflammatory response (adjuvant arthritis).
The comitogen-induced thymocyte proliferation procedure (LAF) was used as o an in vilro measure of tl-e immunos~lpplessive effects of representative compounds.
Briefly, cells from the thymus of normal BALB/c mice are cultured for 7'~ hours with PHA and IL-I 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 halvested and incolporated; radioactivity is determined.
Inhibition of Iymphoproliferation is assessed in percent change in counts per minute from non-drug treated controls. T}-e results are expressed by the following ratio, or as the percent hlhibition of Iymphoproliferation of 1 ~lM.
3H-control thymus cells - H3-rap.lmycin-trellted thymus cells 3H-control thymus cells - H3-test compound-treated cells The results for the rap;mlycin analog (ICsoallalog) alld r.tp;mlycin (ICsorapa) as well as the ratio of the IDs()s of rapamycin to the analog (R/A) are given in the table below. A
ratio less than l.0 means the analog is less potent than rapamycin.
2s The in vivo test procedure is designed to determine the survival time of pinch skin graft from male DBA/2 donors tr;ulsplanted to male BALB/c recipients. The method is adapted from Billingh;ml R.E. and Medawar P.B., J. Exp. Biol. 2~: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 compoulld, intrapelitone.llly. Untleated recipients serve as rejection control.
The graft is monitored daily and observations ale recorded until the graft becomes dry and forms a blackened scab. This is considel-ed as the rejection day. The mean graft survival time (MST -number of days + S.D.) of the drug treatment group is compared with the control group. Rap;lmycill tre.ltlllellt provides a meall graft survival (MST) of 12.0~ 1.7 days.
WO ~a~93~v PCTrUS97/15439 The in vivo adjuvant arthritis standard pharm.lcological test procedure measuresthe ability of test compo-~nds to prevent immune mediated inflammation and inhibit or treat rheumatoid arthritis. The following briefly describes the test procedure used. A
group of rats (male inbred Wistar Lewis rats) are pre-treated with the compound to be s tested (lh prior to antigen) and then injected with Freund's Complete Adjuvant (FCA) in the right hind paw to induce arthritis. The rats are then orally dosed on a Monday, Wednesday, Friday schedule from day 0-14 for a total of 7 doses. Both hind paws are measured on days 1~, 23 and 3(). The difference in paw volume (mL) from day 16 to day 0 is determined and a percent change frolll control is obtained. The left hind paw o (uninjected paw) inflammation is caused by T-cell mediated inflammation and isrecorded as percent change from control. The right hind paw inflammation, on theother hand, is caused by non-specific inflammation. Compounds were tested at a dose of 2 mg/kg. The results ~ue expressed as the percent change in the uninjected paw at day 16 versus control; the more negative the percent change, the more potent thecompound. Rapamycin provides between -7()~o and -90% change versus control, indicating that rapamycin treated rats h,lve between 70-90% less immune-induced inflammation than control rats.
The following table summarizes tlle results of the compounds of this invention in these three standard test procedures.
g W 098/09970 PCTrUS97/15439 Table 1: Summ~try of Pharmacological Test Results EvaluatiolI of ImmulIosllpplessive Activity LA F Skin G raft Adjuvant Arthritis Example ICs() analog _50 rapa R/A MST + S.D. % Change 4.2 0.5 0.12 7.2 0.04 2 2.4 0.00 3 l .47 1 . l 0.75 9.8 0.8 -88 0 3.10 0.8 ().26 9.7 0.5 The results of these standard pharnIacological test procedures demonstrate immunosuppressive activity both in vitro and in vivo for the compounds of this invention. Positive ratios in the LAF test procedure indicate suppression of T cell proliferation. As tralIsplallted pillCh skhI grafts are typically rejected with 6-7 days without the use of an imllIullosupplessive a$elIt, the increased survival time of the skin graft when treated with the compoulIds of this invention further demonstrates their utility as immunosuppressive agents. The reduction of infl~mlmatory joint swelling in the adjuvant rat model demonstrates their utility in the treatment of inflammatory diseases.
}nhibition of growtll of human tumors in VitlO by rapamycin analogs The compound of Example 3 was shown to inhibit in submicromolar concentrations breast (T47D), colon (MIP 1()1), and ovarian (A 2780S) cancer cell lines according to the following assay procedure:
Human tumor cell lines were plated in 96-well plates (250 IlL/well, 1-6 x 104 cells/mL) in RPMI 164() medium, colItaillillg ~'J/o FBS (Fetal Bovine Serum). Twenty-four hours after plating, drugs were added at five log concentrations ~0.01-100 ~g/mL). After 48 hours exposure to dr-lgs, cells were fixed with trichloroacteic acid, and stained with Sulfolhod;tllline B. After washing with trichloroacetic acid, bound dye was solubili~ed in 1() mM Tris base alld Optical Density was detennined using a plate reader. Under conditions of the ass;ly, the optical density is proportional to the number of cells hl the well. IC5()s (concelItr;ltions causing 50% inhibition of cell growth) were detemlilIed fiom the growth hIhibition plo~s. The assay is described in 3s details by Philip SkehalI et al., J. Nation;ll Cancer Institute X2, 1107-1112, 1990.
Based on the results of these stalIdald ph;lrl1lacological test procedures, the compounds are useful in the treatment of transphlntation rejection such as, heart, kidney, liver, bone marlow, and skin tr.lnsplants; autoimmune diseases such as lupus, rheumatoid arthritis, dhlbetes mellitus, myastheni;l gravis, and multiple sclerosis; and , W O 981'~J9;v PCTAUS97/15439 diseases of inflammation such as, psoriasis, dermatitis, eczema, seborrhea, infla,~ laloly bowel disease and pulnlollary inflammation such as asthma, chronic obstructive pulmon.try disease, emphysema, bronchitis and the like; proliferative diseases such as restenosis following angioplasty procedures, and fungal infections.
s The compounds are also expected to be useful in the treatrment of breast, colon, or ovarian cancers in humans.
Pharm~ceutic~1 Composition 0 The compounds may be administered neat or with a pharrnaceutical carrier to a mammal in need thereof. The ph;tml;tceutic;tl carrier may be solid or liquid and the active compound shall be a therapeutic;tlly effective amount.
A solid c;trrier can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, s 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 ingrediellt. In tablets, the active ingredient is mixed with a carrier having the necessary complession properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of 2n the active ingredient. Suitable solid carliers include, for example, calcium phosphate, magnesium ste;uate, talc, sugars, lactose, dextrill, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.
Liquid carriers are used in prep;uing solutions, suspensions, eMulsions, 2s syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutic;tlly acceptable liquid carrier such as water, an organic solvent, a mixture of both or phalmaceutic,llly acceptable oils or fats. The liquid carrier can contain other suitable phamlaceutic.ll additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers Ol osmo-regulators. Suitable examples of liquid carriers for oral and palentelal administration incltlde water (partially containing additives as above, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solutiol1), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and ti1eil deriv;ltives, and oils (e.g. fractionated 3s coconut oil and ar,lchis oil). For palentel;ll admil1istlation, the car~ier can also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are useful W O 981~337~ PCT~US97/15439 in sterile liquid fo~m compositions for p.uentel;ll administr.ltion. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.
Liquid phamlaceutical compositions which are sterile solutions or suspensions s 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 form~ teA
compound can further be administered intr.masally through insuMation of a powderformulation, rectally or vaginally via suppositories, and topically or transdermally.
0 Furthermore, the formulated invention compound can be administered alone or in combination with one or more addidional immunoregluatory agents such as a corticosteroid, cyclophosphamide, r,lpalllyLIcin, cyclosporin A, FK-506, OKT-3 or ATG as established by Stepkowski, Transplantation Proceedings 23: 5()7 (1991).
Preferably, the ph.umaceutical composition is in unit dosage form, e.g. as tablets or capsules. In such foml, the composition is sub-divided in unit doses containing appropriate quantities of the active ingredient; the unit dosage forms can be packaged compositions, for example, packeted powders, vials, ampoules, prefilledsyringes or sachets contaillillg 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 treatmellt must be subjectively deterrnined by the attending physician.
This invention relates to compounds of formula I below or pharm~eutir~l1y acceptable salts thereof which possess immunosuppressive and/or anti tumor and/or s antiinflamrnatory activity in vivo and/or inhibit thymocyte proliferation in vilro. These compounds are therefore useful in the treatment of transplantation rejection, autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, multiple sclerosis, the treatn1ellt of Candicia ~1lbicans infectiolls, treatment of diseases of inflammation neatment of hyperproliferative vascular disease (restenosis) and in the treatrnent of certain human tumors.
BACKGROUND OF THE INVENTION
Rapamycin is a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus, which was follnd to have antifungal activity, particularly against Candida albicans, both in vitro alld in vlvo ~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 (197~); U.S. Patent 3,929,992; and U.S. Patent 3,993,749].
,~- OCH3 4% ~37 26 ,48 8 [~ 2~OH
~0 ~ CH30- ~
HO ~J~4 - 34 46--~ 013 OCH3 5(~ ~ 35 ~ 1 4 2 4; 51 20Rap~mycin (Positions numbered accordillg to Chemical Abstract~s) Rapamycin alone (U.S. Patent 4,885,171) or in combination with picibanil (U.S. Patent 4,401,653) has been shown to have antitumor activity. R. Martel et al.
I
wo g8,~597., rCT/US97/15439 [Can. J. Physiol. Ph~rmacol. 55, 48 (1977)1 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.
s The immunosuppressive effects of rapamycin have been disclosed in FASEB 3, 3411 (19~9). Rapamycin has been shown to be effective in inhibiting transplant rejection (U.S. Patent 5,10(),899). Cyclosporin A and FK-506, other macrocyclic molecules, also have been shown to be effective as immunosuppressive agents, therefore useful in preventing transplant rejection IFASEB 3, 3411 (1989); FASEB 3, o 5256 (1989); and R. Y. Calne et al., Lancet 1183 (1978)]. U. S. patent 5,321,009 discloses a method of prophylactica]ly preventing the onset, preventing the development, and arresting the progression of insulin-dependent diabetes mellitus by administration of rapamycin. U. S. patellt 5,2~,711 discloses a method of preventing or treating hyperproliferative vascular disease by administration of a combination of s rapamycin and heparin. U. S. patent 5,2~6,730 discloses a method of treatingimmunoinflammatory disease by treatlllent with rapamycin alone or in combination with cyclosporin A. U. S. patent 5,2~6,731 provides a method of treating immunoinflammatory bowel disease by administration of rapamycin alone or in combination with cyclosporin A.
Various structural features of rapamycin have been modified in efforts to increase the potency or specificity of phamlacological action. For instance, a number of U. S. patents disclose compounds where one or more of the hydroxy groups having normal stereochemistry at positions 14, 31, and 42 have been converted into acylesters, sulfonyl esters, and carballlates. U. S. patent 5,023,263 discloses 42-oxo 2s rapamycin. V. S. patent 5,258,389 discloses 31 and/or 42 O-alkyl, O-aryl, O-alkenyl, and O-alkynyl ethers of rapamycin having normal stereochemistry at the 42 position.
The PCI' published applicatioll WO 94/~)9()1() discloses 31 and/or 42 O-alkylated rapamycin analogs wherein the keto groups at positions 15 and 33 may be reduced to a hydroxyl group or a methylene group.
SUMMARY OF THE INVENTION
The compounds of this invention are represented by the following chemical formula and are novel. While a number of rapamycin analogs substituted at positions 3s 31 and 42 have been disclosed, substitution at position 29 has not been heretofore disclosed.
W O ~8J~/V PCT~US97/15439 The compounds useful in this invention are represented by the formula below:
~3 W
38 ' ~ - OCH3 3 ~ ~ z~ Z y ~~o ~ CH303 33~
HO~I~4 134 46 ~ 013 OCH3 5( ~ > 35 11 ~ ' ~ 36 4~ 51 wherein W and Y are ORl and X and Z together form a bond or W and X are oR2 and Y and Z together fonn ,t bond, wherein R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O~nCH3 where n is not 1, I O -CH2CH2CH20(CH2CH20)m-CH3, ~(CH2)n-CH2CH(oR3)cH2oR4 where R3 ;~nd R4 are H, Cl-Clo alkyl, or R3 and R4 together ale ethylene, metl~ylene or dimethylmethylene;
~CH2(CH2)n-oR3 where R3 is not H, Cl-Cl() alkyl, or C(O)C1-CIo alkyl, and -CH2(CH2)n-X where X is F, Cl, Br or l;
R2 is selected from H, Cl-Clo alkyl, Ar(CH2)n-, C3-CIo alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH20(CH2CH20)m-CH3, ~CH2(CH2)n-oR3~ -CH2(CH2)n-X
where X is F, Cl, Br or ~;
and -(CH2)n-CH~CH(ORS)CH2OR6 where RS and R6 are selected independently from H, Cl-Cl() alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -CoR7 and -Co2R7~ where R7 is Cl-C6 alkyl, C2-C6 alkellyl, or Ar;
n= 1-10 indepelldelltly: and 2s . _ W O 9~1~3~,~ PCTrUS97/15439 m= 1-5 independently;
In the above definitions of variable substituents, the term alkyl includes straight and branched chain hydrocarbons and the aryl groups (Ar) are selected independently s from phenyl, pyridinyE, quinolinyl, indolyl, furanyl, 1, 2, 3-triazolyl and tetrazolyl.
This invention also encompasses the pharmaceutica]ly acceptable acid addition salts where ehey can be formed.
The compounds of this invention exhibit immunosuppressive and/or antifungal and/or antitumor and/or antiinflammatory activity in vivo and/or inhibit thymocyte o proliferation in vitro and are therefore useful in the ùeatment or inhibition of organ or tissue transplantatioll rejection or host vs. graft disease, proliferative diseases such as restenosis following angioplasty procedures, autoimmune diseases such as lupus, rheumatoid arthritis, diabetes mellitus, and multiple sclerosis; fungal infections, and diseases of inflamm.ltioll such as psoriasis, exzema, sebolThea, inflammatory bowel disease and pulmonary inflammation such ;lS asthma, chronic obstructive pulmonary disease, emphysema, brollchitis and the like. An invention compound was found toinhibit the growth of human breast, colon and ovarian cancer cell lines in submicromolar concentration and it is therefole expected that the invention compounds will be useful in inhibiting these and other tumors in humans.
Detailed Description of the Invention The compounds of this invention can be prepared by standard literature procedures as outlined below. Other suitable bases may be used in place of the 2,6-di-tert-butyl-4-methylpyridine and may include pyridine, lutidine, co]lidine, sodium hydride, or sodiulll carbonate.
f ~0 H ~O R
~ OMe ~~ OMe ~, '2 6 di(t buRty~l) 4-2rnetnylpyridine CH2cl2 ~ I n ¦l SSlr~ ~ ~;0 or ROC(=NH)CCI3, catalytic SSlr ~31 M e O J~ CF3SO3H, benzene o M e O' ~ ~
+ 31-substituted product and 31,42-disubstituted product W O 98/09970 PCTAUS97/lS439 Also, unexpectedly, a product was i.solated (1,3-allyl rearrangement) via triflation at C-28 followed by attack by external nucleophile.
OH ~OH
'J~ OMe ~ OMe , ~2,6-di(t-butyl)-4-methylpyridine ~" ~O R
n 1l CH2CI2 I n ~ 1 SSlr ~31or ROC(=NH)CCI3, catalytic Ss~r ~
M e O ~CF3SO3H, benzene o M e O~ J~~
~ss ~ss The compounds of this invention exhibit immunosuppressive and/or antifungal and/or antitumor and/or antiinflammatory activity in vivo andJor inhibit thymocyte proliferation in vitro and are therefore useful in the treatment of transplantation rejection, autoimmul1e dise~ces (i.e. Iupus, rheumatoid arthritis, diabetes mellitus, multiple sclerosis), Candida al~ica~Ls infections, and diseases of inflammation. An invention compound inhibits in vitro in sub-micron-olar concentrations, the growth of certain human tumor cell lines, including colon (MIP 101), breast (T47D, SKBR-3), and ovarian (A2780S) cells and therefore it is expected that compounds of this inventon will be useful in the treatment of these and other tumors.
The following procedllres are included to exemplify the preparation of inventioncompounds and employ stand.ud labor.ltory techniq~les known to those skilled in the art of organic systhesis.
Example ].
29-O-Ben~,ylI apamycin Toasolution of triflic anhydride (0.19 n-L, 1.13 mmol) in dichloromethane (5.0 rnL) in a round bottom flask (50 mL, flame dried) equipped with a magnetic stirrer at 0 ~C
was added 2,6-di-t-butyl-4-methyl pyridine (().253 g, 1.23 mmol) portionwise. The 2s reaction mixture was deg.lssed, and purged with nitrogen. To the solution was added a solution of benzyl alcohol (().1] mL, 1.06 mmol, diluted in 3 mL dichloromethane) dropwise. The solutioll became a pale white suspensioIl. The reaction was stirred at 0 ~C for 30 min. TLC analysis indicated no residllal benzyl alcohol remained. To the suspension was added another portion of 2,6-di-t-butyl-4-methyl pyridine (0.335 g, 1.63 mmol) portionwise, followed by a solution of rapamycin (0.855 g, 0.94 mmol, in 3 mL dichloromethalle) dropwise. The reaction was stirred at 0 ~C for 30 min, and warmed up to room temperature and stirred overnight. The reaction was quenched with saturated aqueous NaHC03, and the organic and aqueous layers were separated. Iheaqueous layer was extracted three times with ethyl acetate. The organic layers were combined, washed with brine, and dried over sodium sulfate. The solution was filtered and concentrated ln vac~o to afford a pale yellow foam. The product rnixture wasseparated by HPLC (40 % EtOAc/hexane, Dynan1ax 2" silica colun1n, 20 mL/min), and o four fractions were collected. The second fraction which was obtained in 12 % yield was identified as 42-O-benzylrapamycin. The third fraction which was obtained in 14 % yield was identified as the dihydr;lte of the title compound.
H NMR (4()() MHz, DMSO-d6) ~ 7.3~ (m, 5 H, PhCH2O-), 6.]() - 6.42 (m, 4 H, S vinylic), 5.48 (q, 1 H, vinylic), 4.95 (m, 2 H, PhCH20-), 3.32 (s, 3 H, -OC~13), 3.18 (s, 3 H, -OC~3), 3.()5 (s, 3 H, -OCH3); IR (KBr, cm~l): 3420, 2920, 1730, 1645, 1445; MS (neg. FAB): 1()03.4 lMl-, 652, 59(), 349; Anal. Calcd. for Cs3H83Nol3. 2 H2O: C 66.98 %, H 8.56 %, N 1.39 %; Found: C 66.53 %, H 8.51 %, N 1.70 %.
Preparation of substituted alkyl triflates.
To a solution of substituted alkanol (l.()() mmol) in dichloromethane (15.0 mL) in a round bottom flask (25 mL, flame dried) equipped with a magnetic stirrer at roomtemperature was added 2,~-di-t-butyl-4-metllyl pyridine (0.~50 g, 2.92 mmol). The 2s reaction mixture was degassed, purged with nitrogen, and cooled to -78 ~C. To the solution was added dropwise trifluoromethanes-llfonic anhydride (0.170 mL, 0.282 g, 1-.00 mmol) over a period of 5 minutes. The solution became a suspension. The reaction was stirred at () ~C for 3() minutes, then wamled up to room t~lllpe.dlul~. TLC
analysis showed completion of ttle reaction.
General Procedure for the Preparatioll of 42-substituted Derivatives of Rapamycin via Nucleophilic Substitution.
To the solution of substituted alkyl-triflate prep~ared from 1 mmol of alkanol was added at room temperature (unless otherwise noted) rap~mycin (914 mg, 1 mmol). The mixture was stirred between 4 and 12() h (monitoring of the extent of the reaction was ~, wo ~ 3~7~ PCT/USg7/I5439 carried out by TLC). When desired convelsioll has been achieved the reaction wasquenched with saturated aqueous NaHCO3, and the organic and aqueous layers were separated. The aqueous layer was extracted three times with ethyl acetate. The organic layers were combined, washed with brine, and dried over sodium sulfate. The solution s was filtered and concen~ated i~ vacuo to afford a product. Pure products were isolated by HPLC (norrnal phase- Dynamax 2" silica column, eluent EtOAc: hexane,20 mL4min; reversed ph~se -Dynamax 2" C18 column, eluent MeCN: water, 20 mL/min. Spectroscopic analyses were used to confirm the structures.
0 Example 2.
42-0-(4-chlorobutyl )rapamycin Method A. Alkanol used: 1()8 mg (I mmol) of l-chloro-butane-4-ol. Reaction tirne:
S 72 h at 25 ~C. Separatioll tech~ u~ en1ployed: 2" Dynal~ x silica column, eluent 40%
EtOAc: hexane,20 mL/min. Yield of product: 18() mg (18 %).
Spectral data follows: IH NMR (400 MHz, DMSO-d6 ~ 6.08-6.45 (m, 4 H), 3.4-3.7 (m, 8 H), 3.1S (s, 3H), 3.04 (s, 3 H). IR (KBr, cm~l) 3420, 2930, 1715, 1650, 1620, 1460. MS (neg. FAB) 1003.4 ~MI-, 590.2, 411.2. Anal. calcd. for CssHg6NO13CI: C 65.75 %, H 8.63 %, N 1.39 %; Found: C 65.14 %, H 8.64 %, N 1.18 %.
Example 3.
2s 42-0-(2-[2-(2-methoxy etho~y)-ethoxy]-ethyl)-l apamycin Method A. Alkanol used: 165 mg (I mmol) of triethylenegycol monomethyl ether.
Reaction time: 12() h ~I 1() ~C. Separation techni~lue employed: 2" Dynamax silica column, eluent 7()~c EtOAc: hexane,2() mL/min. Yield of product: 210 mg (19 %).
Spectral data follows: ~H NMR (4()() MHz, DMSO-d6 ~ 6.()5-6.45 (m, 4 H), 3.4-3.65 (m, 12 H), 3.23 (s, 3 H), 3.14 (s, 3 H), 3.()4 (s, 3 H). IR (KBr, cm~l ) 3420, 2925, 1720, 1645, 145(); MS (neg. FAB) 1()59.4 IM]-, 590.2, 467.2. Anal. Calcd.
3s for C5XHg3NOl6. H~O C 64.6() %, H 8.81 %, N 1.28 %; Found: C 63.96 %, H
8.64 %, N 1.15 %.
W O 9~ 7v PCT~US97/15439 Pharmacolo~y Immunosuppressive activity was evalu~ted in an in vitro standard s pharmacological test procedllre to measure Iymphocyte proliferation (LAF), in an in vivo procedure to evaluate the survival time of a pinch skin graft, and in an in vivo procedure to determine inhibition of T-cell mediated inflammatory response (adjuvant arthritis).
The comitogen-induced thymocyte proliferation procedure (LAF) was used as o an in vilro measure of tl-e immunos~lpplessive effects of representative compounds.
Briefly, cells from the thymus of normal BALB/c mice are cultured for 7'~ hours with PHA and IL-I 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 halvested and incolporated; radioactivity is determined.
Inhibition of Iymphoproliferation is assessed in percent change in counts per minute from non-drug treated controls. T}-e results are expressed by the following ratio, or as the percent hlhibition of Iymphoproliferation of 1 ~lM.
3H-control thymus cells - H3-rap.lmycin-trellted thymus cells 3H-control thymus cells - H3-test compound-treated cells The results for the rap;mlycin analog (ICsoallalog) alld r.tp;mlycin (ICsorapa) as well as the ratio of the IDs()s of rapamycin to the analog (R/A) are given in the table below. A
ratio less than l.0 means the analog is less potent than rapamycin.
2s The in vivo test procedure is designed to determine the survival time of pinch skin graft from male DBA/2 donors tr;ulsplanted to male BALB/c recipients. The method is adapted from Billingh;ml R.E. and Medawar P.B., J. Exp. Biol. 2~: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 compoulld, intrapelitone.llly. Untleated recipients serve as rejection control.
The graft is monitored daily and observations ale recorded until the graft becomes dry and forms a blackened scab. This is considel-ed as the rejection day. The mean graft survival time (MST -number of days + S.D.) of the drug treatment group is compared with the control group. Rap;lmycill tre.ltlllellt provides a meall graft survival (MST) of 12.0~ 1.7 days.
WO ~a~93~v PCTrUS97/15439 The in vivo adjuvant arthritis standard pharm.lcological test procedure measuresthe ability of test compo-~nds to prevent immune mediated inflammation and inhibit or treat rheumatoid arthritis. The following briefly describes the test procedure used. A
group of rats (male inbred Wistar Lewis rats) are pre-treated with the compound to be s tested (lh prior to antigen) and then injected with Freund's Complete Adjuvant (FCA) in the right hind paw to induce arthritis. The rats are then orally dosed on a Monday, Wednesday, Friday schedule from day 0-14 for a total of 7 doses. Both hind paws are measured on days 1~, 23 and 3(). The difference in paw volume (mL) from day 16 to day 0 is determined and a percent change frolll control is obtained. The left hind paw o (uninjected paw) inflammation is caused by T-cell mediated inflammation and isrecorded as percent change from control. The right hind paw inflammation, on theother hand, is caused by non-specific inflammation. Compounds were tested at a dose of 2 mg/kg. The results ~ue expressed as the percent change in the uninjected paw at day 16 versus control; the more negative the percent change, the more potent thecompound. Rapamycin provides between -7()~o and -90% change versus control, indicating that rapamycin treated rats h,lve between 70-90% less immune-induced inflammation than control rats.
The following table summarizes tlle results of the compounds of this invention in these three standard test procedures.
g W 098/09970 PCTrUS97/15439 Table 1: Summ~try of Pharmacological Test Results EvaluatiolI of ImmulIosllpplessive Activity LA F Skin G raft Adjuvant Arthritis Example ICs() analog _50 rapa R/A MST + S.D. % Change 4.2 0.5 0.12 7.2 0.04 2 2.4 0.00 3 l .47 1 . l 0.75 9.8 0.8 -88 0 3.10 0.8 ().26 9.7 0.5 The results of these standard pharnIacological test procedures demonstrate immunosuppressive activity both in vitro and in vivo for the compounds of this invention. Positive ratios in the LAF test procedure indicate suppression of T cell proliferation. As tralIsplallted pillCh skhI grafts are typically rejected with 6-7 days without the use of an imllIullosupplessive a$elIt, the increased survival time of the skin graft when treated with the compoulIds of this invention further demonstrates their utility as immunosuppressive agents. The reduction of infl~mlmatory joint swelling in the adjuvant rat model demonstrates their utility in the treatment of inflammatory diseases.
}nhibition of growtll of human tumors in VitlO by rapamycin analogs The compound of Example 3 was shown to inhibit in submicromolar concentrations breast (T47D), colon (MIP 1()1), and ovarian (A 2780S) cancer cell lines according to the following assay procedure:
Human tumor cell lines were plated in 96-well plates (250 IlL/well, 1-6 x 104 cells/mL) in RPMI 164() medium, colItaillillg ~'J/o FBS (Fetal Bovine Serum). Twenty-four hours after plating, drugs were added at five log concentrations ~0.01-100 ~g/mL). After 48 hours exposure to dr-lgs, cells were fixed with trichloroacteic acid, and stained with Sulfolhod;tllline B. After washing with trichloroacetic acid, bound dye was solubili~ed in 1() mM Tris base alld Optical Density was detennined using a plate reader. Under conditions of the ass;ly, the optical density is proportional to the number of cells hl the well. IC5()s (concelItr;ltions causing 50% inhibition of cell growth) were detemlilIed fiom the growth hIhibition plo~s. The assay is described in 3s details by Philip SkehalI et al., J. Nation;ll Cancer Institute X2, 1107-1112, 1990.
Based on the results of these stalIdald ph;lrl1lacological test procedures, the compounds are useful in the treatment of transphlntation rejection such as, heart, kidney, liver, bone marlow, and skin tr.lnsplants; autoimmune diseases such as lupus, rheumatoid arthritis, dhlbetes mellitus, myastheni;l gravis, and multiple sclerosis; and , W O 981'~J9;v PCTAUS97/15439 diseases of inflammation such as, psoriasis, dermatitis, eczema, seborrhea, infla,~ laloly bowel disease and pulnlollary inflammation such as asthma, chronic obstructive pulmon.try disease, emphysema, bronchitis and the like; proliferative diseases such as restenosis following angioplasty procedures, and fungal infections.
s The compounds are also expected to be useful in the treatrment of breast, colon, or ovarian cancers in humans.
Pharm~ceutic~1 Composition 0 The compounds may be administered neat or with a pharrnaceutical carrier to a mammal in need thereof. The ph;tml;tceutic;tl carrier may be solid or liquid and the active compound shall be a therapeutic;tlly effective amount.
A solid c;trrier can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, s 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 ingrediellt. In tablets, the active ingredient is mixed with a carrier having the necessary complession properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of 2n the active ingredient. Suitable solid carliers include, for example, calcium phosphate, magnesium ste;uate, talc, sugars, lactose, dextrill, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.
Liquid carriers are used in prep;uing solutions, suspensions, eMulsions, 2s syrups, elixirs and pressurized compositions. The active ingredient can be dissolved or suspended in a pharmaceutic;tlly acceptable liquid carrier such as water, an organic solvent, a mixture of both or phalmaceutic,llly acceptable oils or fats. The liquid carrier can contain other suitable phamlaceutic.ll additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers Ol osmo-regulators. Suitable examples of liquid carriers for oral and palentelal administration incltlde water (partially containing additives as above, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solutiol1), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and ti1eil deriv;ltives, and oils (e.g. fractionated 3s coconut oil and ar,lchis oil). For palentel;ll admil1istlation, the car~ier can also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are useful W O 981~337~ PCT~US97/15439 in sterile liquid fo~m compositions for p.uentel;ll administr.ltion. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.
Liquid phamlaceutical compositions which are sterile solutions or suspensions s 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 form~ teA
compound can further be administered intr.masally through insuMation of a powderformulation, rectally or vaginally via suppositories, and topically or transdermally.
0 Furthermore, the formulated invention compound can be administered alone or in combination with one or more addidional immunoregluatory agents such as a corticosteroid, cyclophosphamide, r,lpalllyLIcin, cyclosporin A, FK-506, OKT-3 or ATG as established by Stepkowski, Transplantation Proceedings 23: 5()7 (1991).
Preferably, the ph.umaceutical composition is in unit dosage form, e.g. as tablets or capsules. In such foml, the composition is sub-divided in unit doses containing appropriate quantities of the active ingredient; the unit dosage forms can be packaged compositions, for example, packeted powders, vials, ampoules, prefilledsyringes or sachets contaillillg 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 treatmellt must be subjectively deterrnined by the attending physician.
Claims (11)
1. A compound having the formula:
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n-X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3, -CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n = 1-10 independently;
m = 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n-X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3, -CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n = 1-10 independently;
m = 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
2. A compound according to claim 1 which is 29-O-benzylrapamycin.
3. A compound according to claim 1 which is 42-O-(4-chlorobutyl)rapamycin.
4. A compound according to claim 1 which is 42-O-(2-[2-(2-methoxyethoxy)-ethoxylethyl)rapamycin
5. A method of treating transplantation rejection or host vs. graft disease in amammal by administering thereto an immunosuppressing effective amount of a compound having the formula:
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n -Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n -CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n -X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3, -CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one call be formed.
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n -Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n -CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n -X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3, -CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one call be formed.
6. A method of treating asthma in a mammal by administering thereto an asthma inhibiting effective amount of a compound having the formula wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n -Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n -CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n -X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3, -CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkellyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
R1 is selected from -(CH2)n -Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n -CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n -X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3, -CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkellyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
7. A method of treating rheumatoid arthritis in a mammal by administering thereto an arthritis inhibiting effective amount of a compound having the formula:
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n -Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n -CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n -X whele X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3,-CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n -Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n -CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n -X whele X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3,-CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
8. A method of treating fungal infections in a mammal by administering thereto an antifungal effective amount of a compound having the formula:
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n -Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n -CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n -X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3, -CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n -Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n -CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n -OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n -X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)n CH3, -CH2CH2CH2O(CH2CH2O)m -CH3, -CH2(CH2)n -OR3, -CH2(CH2)n -X
where X is F, Cl, Br or I;
and -(CH2)n -CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n -Ar, -CONH(CH2)n -Ar or COC(CH3)2-(CH2)n -Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1,2,3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
9. A method of inhibiting restenosis in a mammal by administering thereto a restenosis inhibiting effective amount of a compound having the formula:
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n-OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n-X where X is F, Cl, Br or 1;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR3, -CH2(CH2)n-X
where X is F, Cl, Br or I;
and -(CH2)n-CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR7 and -CO2R7, where R7 is C1-6 alkyl, C2-6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1, 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)n CH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n-OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n-X where X is F, Cl, Br or 1;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR3, -CH2(CH2)n-X
where X is F, Cl, Br or I;
and -(CH2)n-CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR7 and -CO2R7, where R7 is C1-6 alkyl, C2-6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1, 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
10. A method of treating breast cancer, colon cancer, or ovarian cancer which comprises administration to a person having such a cancer of a cancer-inhibitingamount of a compound having the formula:
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)nCH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n-OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n-X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR3, -CH2(CH2)n-X
where X is F, Cl, Br or I;
and -(CH2)n-CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1, 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)nCH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m-CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n-OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n-X where X is F, Cl, Br or I;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR3, -CH2(CH2)n-X
where X is F, Cl, Br or I;
and -(CH2)n-CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1, 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
11. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of the formula:
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)nCH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n-OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n-X where X is F, Cl, Br or l;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR3, -CH2(CH2)n-X
where X is F, Cl, Br or 1;
and -(CH2)n,-CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1, 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
wherein W and Y are OR1 and X and Z together form a bond or W and X are OR2 and Y and Z together form a bond, wherein:
R1 is selected from -(CH2)n-Ar where Ar is not phenyl, -(CH2CH2O)nCH3 where n is not 1, -CH2CH2CH2O(CH2CH2O)m -CH3, -(CH2)n-CH2CH(OR3)CH2OR4 where R3 and R4 are H, C1-C10 alkyl, or R3 and R4 together are ethylene, methylene or dimethylmethylene;
-CH2(CH2)n-OR3 where R3 is not H, C1-C10 alkyl, or C(O)C1-C10 alkyl;
and -CH2(CH2)n-X where X is F, Cl, Br or l;
R2 is selected from H, C1-C10 alkyl, Ar(CH2)n-, C3-C10 alkenyl, -(CH2CH2O)nCH3, -CH2CH2CH2O(CH2CH2O)m-CH3, -CH2(CH2)n-OR3, -CH2(CH2)n-X
where X is F, Cl, Br or 1;
and -(CH2)n,-CH2CH(OR5)CH2OR6 where R5 and R6 are selected independently from H, C1-C10 alkyl, -(CH2)n-Ar, -CONH(CH2)n-Ar or COC(CH3)2-(CH2)n-Ar, -COR7 and -CO2R7, where R7 is C1-C6 alkyl, C2-C6 alkenyl, or Ar;
n= 1-10 independently;
m= 1-5 independently; and Ar is selected independently from phenyl, pyridinyl, quinolinyl, indolyl, furanyl;
1, 2, 3-triazolyl and tetrazolyl, and a pharmaceutically acceptable acid addition salt where one can be formed.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US70959196A | 1996-09-09 | 1996-09-09 | |
| US08/709,591 | 1996-09-09 | ||
| PCT/US1997/015439 WO1998009970A2 (en) | 1996-09-09 | 1997-09-03 | Alkylated rapamycin derivatives |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2266039A1 true CA2266039A1 (en) | 1998-03-12 |
Family
ID=24850507
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002266039A Abandoned CA2266039A1 (en) | 1996-09-09 | 1997-09-03 | Alkylated rapamycin derivatives |
Country Status (6)
| Country | Link |
|---|---|
| EP (1) | EP0927182A2 (en) |
| JP (1) | JP2001500126A (en) |
| CN (1) | CN1235608A (en) |
| AU (1) | AU4176897A (en) |
| CA (1) | CA2266039A1 (en) |
| WO (1) | WO1998009970A2 (en) |
Families Citing this family (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6890546B2 (en) | 1998-09-24 | 2005-05-10 | Abbott Laboratories | Medical devices containing rapamycin analogs |
| US6015815A (en) | 1997-09-26 | 2000-01-18 | Abbott Laboratories | Tetrazole-containing rapamycin analogs with shortened half-lives |
| TW557297B (en) * | 1997-09-26 | 2003-10-11 | Abbott Lab | Rapamycin analogs having immunomodulatory activity, and pharmaceutical compositions containing same |
| US7455853B2 (en) | 1998-09-24 | 2008-11-25 | Abbott Cardiovascular Systems Inc. | Medical devices containing rapamycin analogs |
| EP2783686B1 (en) | 2001-02-19 | 2017-06-21 | Novartis AG | Combination of a rapamycin derivative and letrozole for treating breast cancer |
| DE60206512T2 (en) | 2001-08-22 | 2006-06-22 | Wyeth | RAPAMYCIN 29-ENOLE |
| US6680330B2 (en) | 2001-08-22 | 2004-01-20 | Wyeth | Rapamycin dialdehydes |
| KR101004224B1 (en) | 2002-02-01 | 2010-12-27 | 어리어드 파마슈티칼스, 인코포레이티드 | Phosphorus-containing compounds & uses thereof |
| WO2004060283A2 (en) | 2002-12-16 | 2004-07-22 | Nitromed, Inc. | Nitrosated and nitrosylated rapamycin compounds, compositions and methods of use |
| JP4716280B2 (en) * | 2004-03-01 | 2011-07-06 | テルモ株式会社 | Process for producing O-alkylated rapamycin derivatives |
| EP2583678A3 (en) | 2004-06-24 | 2013-11-13 | Novartis Vaccines and Diagnostics, Inc. | Small molecule immunopotentiators and assays for their detection |
| CN104854112A (en) * | 2012-11-30 | 2015-08-19 | 杭州归创生物医药有限公司 | Rafamycin analogs and methods for making same |
| US20160130280A1 (en) * | 2013-06-20 | 2016-05-12 | Novartis Ag | Alkylation with an alkyl fluoroalkyl sulfonate |
| HUE067446T2 (en) | 2018-06-15 | 2024-10-28 | Janssen Pharmaceutica Nv | Rapamycin analogs and uses thereof |
| EP4069223A4 (en) | 2019-12-05 | 2023-12-20 | Janssen Pharmaceutica NV | Rapamycin analogs and uses thereof |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5151413A (en) * | 1991-11-06 | 1992-09-29 | American Home Products Corporation | Rapamycin acetals as immunosuppressant and antifungal agents |
| ZA935111B (en) * | 1992-07-17 | 1994-02-04 | Smithkline Beecham Corp | Rapamycin derivatives |
| ZA935112B (en) * | 1992-07-17 | 1994-02-08 | Smithkline Beecham Corp | Rapamycin derivatives |
| GB9221220D0 (en) * | 1992-10-09 | 1992-11-25 | Sandoz Ag | Organic componds |
| US5258389A (en) * | 1992-11-09 | 1993-11-02 | Merck & Co., Inc. | O-aryl, O-alkyl, O-alkenyl and O-alkynylrapamycin derivatives |
| GB9307491D0 (en) * | 1993-04-08 | 1993-06-02 | Sandoz Ltd | Organic compounds |
-
1997
- 1997-09-03 CN CN97199423A patent/CN1235608A/en active Pending
- 1997-09-03 WO PCT/US1997/015439 patent/WO1998009970A2/en not_active Application Discontinuation
- 1997-09-03 AU AU41768/97A patent/AU4176897A/en not_active Abandoned
- 1997-09-03 EP EP97939749A patent/EP0927182A2/en not_active Withdrawn
- 1997-09-03 JP JP10512815A patent/JP2001500126A/en active Pending
- 1997-09-03 CA CA002266039A patent/CA2266039A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| JP2001500126A (en) | 2001-01-09 |
| WO1998009970A2 (en) | 1998-03-12 |
| CN1235608A (en) | 1999-11-17 |
| WO1998009970A3 (en) | 1998-04-16 |
| AU4176897A (en) | 1998-03-26 |
| EP0927182A2 (en) | 1999-07-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5922730A (en) | Alkylated rapamycin derivatives | |
| US5023263A (en) | 42-oxorapamycin | |
| US5100883A (en) | Fluorinated esters of rapamycin | |
| AU641319B2 (en) | Amide esters of rapamycin | |
| US5233036A (en) | Rapamycin alkoxyesters | |
| US5221670A (en) | Rapamycin esters | |
| US5378696A (en) | Rapamycin esters | |
| EP0666861B1 (en) | Rapamycin arylcarbonyl and alkoxycarbonyl carbamates as immunosuppressive and antifungal agents | |
| US5256790A (en) | 27-hydroxyrapamycin and derivatives thereof | |
| CA2067541C (en) | Reduction products of rapamycin | |
| US5130307A (en) | Aminoesters of rapamycin | |
| RU2143434C1 (en) | Derivatives of rapamycin and pharmaceutical composition | |
| US5164399A (en) | Rapamycin pyrazoles | |
| US5120842A (en) | Silyl ethers of rapamycin | |
| KR100444003B1 (en) | Water-soluble rapamycin esters and pharmaceutical compositions comprising them | |
| AU639819B2 (en) | Hydrogenated rapamycin derivatives | |
| US6432973B1 (en) | Water soluble rapamycin esters | |
| AU653175B2 (en) | Carboxylic acid esters of rapamycin | |
| CA2266039A1 (en) | Alkylated rapamycin derivatives | |
| US5358944A (en) | Rapamycin esters for treating transplantation rejection | |
| WO1998009972A1 (en) | Rapamycin derivatives with unnatural stereochemistries | |
| SK281787B6 (en) | Rapamycin hydroxyesters, process for their preparation and pharmaceutical compositions containing them | |
| WO1993005046A1 (en) | Aminodiesters of rapamycin | |
| EP0467606A1 (en) | Rapamycin derivatives | |
| AU647616B2 (en) | Reduction products of rapamycin as immunosuppressants, antiinflammatory or antifungal agents |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Dead |