`:
`5,100,883
`[11] Patent Number:
`United States Patent
`Schiehser
`[45] Date of Patent: Mar. 31, 1992
`
`
`[19]
`
`(54) FLUORINATED ESTERS OF RAPAMYCIN
`
`(75]
`
`Inventor: Guy A. Schiehser, Yardley, Pa.
`
`{73] Assignee: American Home Products
`Corporation, New York, N.Y.
`
`[21] Appl. No.: 682,793
`
`[22] Filed:
`
`Apr. 8, 1991
`
`(Sil Ent. CS wo. A61K 35/74; CO7D 491/14
`[$2] U.S. Ch. wees §14/183; 514/321;
`540/456
`[58] Field of Search ................. 540/456; 514/183, 321
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`3,929,992 12/1975 Sehgal et al. oe 424/122
`
`3,993,749 11/1976 Sehgal et al.
`ee $24/122
`2/1982 Rakhit ......cssscsssssessenseeen 424/122
`4,316,885
`
`ceceseeseeeenceeee 424/114
`4,401,653
`8/1983 Eng oo
`3/1987 Stella et al. ......
`vee 546/90
`4,650,803
`4,885,171 12/1989 Surendra et al. oe 424/122
`
`OTHER PUBLICATIONS
`
`J. Antibiot. 28, 721-726, 727-732 (1975).
`J. Antibiot. 31, 539-545 (1978).
`Can. J. Physiol. Pharmacol. 55, 48 (1977).
`FASEB3, 3411, 5256 (1989).
`Lancet pp. 1183-1185 (1978).
`Med. Sci. Res. 17:877 (1989).
`
`Primary Examiner—Robert T. Bond
`Attorney, Agent, or Firm—Arnold S. Milowsky
`
`[57]
`
`ABSTRACT
`
`A compound ofthe structure
`
`oR!
`
`OMe
`
`.
`
`—
`
`
`
`Ox
`HO
`
`Wherein
`:
`2
`1
`R' and R* are each,independently, hydrogen or
`
`1
`CR},
`
`R3 is a mono-, di-, poly-, or per-fluorinated alkyl
`group with the proviso that R! and R2are both not
`hydrogen, which by virtue of its immunosuppres-
`sive activity is useful
`in treating transplantation
`rejection, host vs. graft disease, autoimmune dis-
`eases and diseases of inflammation; by virtue ofits
`antitumoractivity is useful in treating solid tumors;
`and by virtue of its antifungal activity is useful in
`treating fungal infections.
`
`11 Claims, No Drawings
`
`West-Ward Pharm.
`Exhibit 1043
`Page 001
`
`West-Ward Pharm.
`Exhibit 1043
`Page 001
`
`
`
`1
`
`FLUORINATED ESTERS OF RAPAMYCIN
`
`5,100,883
`
`2
`R! and R2 are each, independently, hydrogen or
`
`BACKGROUNDOF THE INVENTION
`
`This invention relates to novel esters of rapamycin
`and a method for using them in the treatment oftrans-
`plantation rejection, host vs. graft disease, autoimmune
`diseases, diseases of inflammation, solid tumors, and
`fungal infections.
`Rapamycin is a macrocyclic triene antibiotic pro-
`duced by Streptomyces hygroscopicus, which was found
`to have antifungal activity, particularly against Candida
`albicans, both in vitro and in vivo [C. Vezinaetal., J.
`Antibiot. 28, 721 (1975); S. N. Sehgal et al., J. Antibiot.
`28, 727 (1975); H. A. Baker et al., J. Antibiot. 31, 539
`(1978); U.S. Pat. No. 3,929,992; and U.S. Pat. No.
`3,993,749].
`Rapamycin alone (U.S. Pat. No. 4,885,171) or in com-
`bination with picibanil (U.S. Pat. No. 4,401,653) has
`been shownto have antitumoractivity. R. Martel et al.
`{Can. J. Physiol. Pharmacol. 55, 48 (1977)] disclosed
`that rapamycin is effective in the experimental allergic
`encephalomyelitis model, a mode! 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). Cyclosporin A
`and FK-506, other macrocyclic molecules, also have
`been shown to be effective as immunosuppressive
`agents, therefore useful in preventing transplant rejec-
`tion [FASEB3, 3411 (1989); FASEB 3, 5256 (1989); and
`R. Y. Calne et al., Lancet 1183 (1978)].
`Mono-and diacylated derivatives of rapamycin (es-
`terified at the 28 and 43 positions) have been shown to
`be useful as antifungal agents (U.S. Pat. No. 4,316,885)
`and used to make watersoluble prodrugs of rapamycin
`(U.S. Pat. No. 4,650,803). Recently,
`the numbering
`convention for rapamycin has been changed; therefore
`according to Chemical Abstracts nomenclature,
`the
`esters described above would beat the 31- and 42-posi-
`tions.
`
`DESCRIPTION OF THE INVENTION
`
`This invention provides derivatives of rapamycin
`which are useful as immunosuppressive, anti-inflamma-
`tory, and antifungal agents having the structure
`
`or!
`
`
`
`wherein
`
`i
`—CR?.
`
`10
`
`R3 is a mono-, di-, poly-, or per-fluorinated alkyl
`group of 1-10 carbon atoms; with the proviso that
`R! and R2 are both not hydrogen.
`Of these compounds, preferred members are those in
`which R?is hydrogen;those in which R3is a mono-, di-,
`poly-, or per-fluorinated alkyl group of 1-6 carbon
`atoms; and those in which R? is hydrogen and R3 is a
`mono-, di-, poly-, or per-fluorinated alkyl group of 1-6
`carbon atoms.
`The compounds of this invention acylated at the
`42-position can be prepared by acylating rapamycin
`with an acylating agent having the general structure
`
`W
`XCR3
`
`where X is OH,in the presence of a coupling reagent,
`such as CMC (1-cyclohexyl-3-(2-morpholinoethyl)car-
`bodimide metho-para-toluenesulphonate). The com-
`poundsof this invention also can be prepared by using
`an anhydride of the above described carboxylic acid as
`the acylating species. In addition, the acylating species
`can be an acid halide, where X can be Cl, Br, or I.
`Alternatively,
`reagents such as Ishikawa’s Reagent
`(N,N-diethy!-1,1-2,3,3,3-hexafluoropropylamine)
`can
`be used as an acylating reagent to give compounds of
`this invention.
`The compoundsofthis invention acylated at both the
`31- and 42-positions can be prepared by the methods
`described abovebyincreasing variables such as reaction
`time, temperature, and quantity of acylating agent.
`The 31-acylated compoundsof this invention can be
`prepared by protecting the 42-alcohol of rapamycin
`with a protecting group, such as with a tert-butyl dime-
`thylsilyl group in the presence of a base, such as imidaz-
`ole, followed by acylation of the 31-position with an
`acylating agent having the general structure shown
`above. Removal of the protecting group provides the
`31-acylated compounds. In the case of the tert-butyl
`dimethylsily] protecting group, deprotection can be
`accomplished under mildly acidic conditions, such as
`with a mixture of aqueous acetic acid and THF.
`Having the 31-position acylated and the 42-position
`deprotected, the 42-position can be reacted with a dif-
`ferent acylating agent than was reacted with the 31-
`alcohol, to give compounds having different acyl moi-
`eties at the 31- and 42-positions. Alternatively, the 42-
`acyl compounds, prepared as described above, can be
`reacted with an acylating agent having a different struc-
`ture to provide compoundshaving different acy] moi-
`eties at the 31- and 42-positions.
`The acylating groups used to prepare the compounds
`of the invention are commercially available or can be
`prepared by methodsthat are disclosed in the literature.
`Immunosuppressive activity was evaluated in an in
`vitro standard pharmacological test procedure to mea-
`sure lymphocyte proliferation (LAF) and in two in vivo
`standard pharmacological test procedures. Thefirst in
`vivo procedure was a popliteal lymph node (PLN) test
`procedure which measured the effect of compoundsof
`this invention on a mixed lymphocyte reaction and the
`
`30
`
`35
`
`45
`
`50
`
`55
`
`65
`
`West-Ward Pharm.
`Exhibit 1043
`Page 002
`
`West-Ward Pharm.
`Exhibit 1043
`Page 002
`
`
`
`5,100,883
`
`4
`The following table summarizes the results of repre-
`sentative compounds of this invention in these three
`standard test procedures.
`TABLE1
`Skin Graft
`LAF*
`PLN*
`(days + SD)
`(ratio)
`(ratio)
`Compound
`10.33 + 1.0
`40
`1.28
`Example |
`12.0 + 1.3
`2.3
`1.03
`Example 2
`+
`27
`0.23
`Example 3
`+
`14.8
`0.96
`Example 4
`
`Rapamycin 12.0 + 1.7 1 1
`
`
`*Calculation of ratios was described supra.
`+ Not evaluated
`
`3
`second in vivo procedure evaluated the survival time of
`a pinch skin graft.
`The comitugen-induced thymocyte proliferation pro-
`cedure (LAF) was used as an in vitro measure of the
`immunosuppressive effects of
`representative com-
`pounds. Briefly, cells from the thymus of normal
`BALB/c miceare cultured for 72 hours with PHA and
`IL-1 and pulsed with tritiated thymidine duringthelast
`six hours. Cells are cultured with and without various
`concentrations of rapamycin, cyclosporin A, or test
`compound. Cells are harvested and incorporated radio-
`activity is determined. Inhibition of lymphoprolifera-
`tion is assessed as percent change in counts per minute
`from non-drug treated controls. The results are ex-
`pressed by the following ratio, or as the percent inhibi-
`tion of lymphoproliferation at 1 uM.
`
`3H-contro} thymus cells — H}-rapamycin-treated thymuscells
`3H-control thymus cells — H}-test compound-treated cells
`
`20
`
`A mixed lymphocyte reaction (MLR) occurs when
`lymphoid cells from genetically distinct animals are
`combinedin tissue culture. Each stimulates the other to
`undergoblast transformation whichresults in increased
`DNAsynthesis that can be quantified by the incorpora-
`tion oftritiated thymidine. Since stimulating a MLRisa
`function of disparity at Major Histocompatibility anti-
`gens,an in vivo popliteal lymph node (PLN)test proce-
`dure closely correlates to host vs. graft disease. Briefly,
`irradiated spleen cells from BALB/c donors are in-
`jected 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 Day4, tritiated thymidineis giveni-p.,
`b.i.d. On Day 5, the hind popliteal lymph nodes are
`removed anddissolved, and radioactivity counted. The
`corresponding left PLN serves as the control for the
`PLNfrom the injected hind foot. Percent suppression is
`calculated using the non-drug treated animals as allo-
`genic 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
`bythe following ratio:
`
`3H-PLNcells contro! C3H mouse —
`*H-PLN cells rapamycin-treated C3H mouse
`H-PLNcells 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 MedawarP. B., J. Exp. Biol. 28:385-402, (1951).
`Briefly, a pinch skin graft from the donoris 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 concentra-
`tions of cyclosporin A as test control or the test com-
`pound, intraperitoneally. Untreated recipients serve as
`rejection control. The graft is monitored daily and ob-
`servations are recorded until the graft becomes dry and
`forms a blackened scab. This is considered as the rejec-
`tion day. The mean graft survival
`time (number of
`days+S.D.) of the drug treatment group is compared
`with the control group.
`
`35
`
`40
`
`45
`
`50
`
`60
`
`65
`
`The results of these standard pharmacological test
`procedures demonstrate immunosuppressive activity
`both in vitro and in vivo for the compounds ofthis
`invention. Positive ratios in the LAF and PLNtest
`procedures indicate suppression of T cell proliferation.
`As a transplanted pinch skin grafts are typically re-
`jected within 6-7 days without the use of an immuno-
`suppressive agent, the increased survival time of the
`skin graft when treated with the compounds ofthis
`invention further demonstrates their utility as immuno-
`suppressive agents.
`Because the compoundsofthis invention are structur-
`ally similar to rapamycin and have a similar activity
`profile to rapamycin, the compoundsofthis invention
`also are considered to have antitumor and antifungal
`activities.
`Based on the results of these standard pharmacologi-
`cal test procedures, the compoundsare useful
`in the
`treatment of transplantation rejection such as, heart,
`kidney, liver, bone marrow,andskin transplants; auto-
`immune diseases such as, lupus, rheumatoid arthritis,
`diabetes mellitus, myasthenia gravis, and multiple scle-
`rosis; and diseases of inflammation such as, psoriasis,
`dermatitis, eczema, seborrhea,
`inflammatory bowel
`disease; solid tumors; 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 mayalso act as flavoring agents, lubricants, solu-
`bilizers, suspending agents,fillers, glidants, compression
`aids, binders or tablet-disintegrating agents; it can also
`be an encapsulating material. In powders, the carrieris
`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% ofthe active
`ingredient. Suitable solid carriers include, for example,
`calcium phosphate, magnesium stearate,
`talc, sugars,
`lactose, dextrin, starch, gelatin, cellulose, methy] cellu-
`lose, sodium carboxymethy! cellulose, polyvinylpyr-
`rolidine, low melting waxes and ion exchangeresins.
`Liquid carriers are used in preparing solutions, sus-
`pensions, emulsions, syrups, elixirs and pressurized
`compositions. The active ingredient can be dissolved or
`suspended in a pharmaceutically acceptable liquid car-
`rier such as water, an organic solvent, a mixture of both
`or pharmaceutically acceptable oils or fats. The liquid
`carrier can contain other suitable pharmaceutical addi-
`tives such as solubilizers, emulsifiers, buffers, preserva-
`tives, sweeteners, flavoring agents, suspending agents,
`thickening agents, colors, viscosity regulators, stabiliz-
`
`West-Ward Pharm.
`Exhibit 1043
`Page 003
`
`West-Ward Pharm.
`Exhibit 1043
`Page 003
`
`
`
`5
`ers or osmo-regulators. Suitable examples of liquid car-
`riers for oral and parenteral administration include
`water (partially containing additives as above,e.g. cel-
`Julose derivatives, preferably sodium carboxymethy]
`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 isopro-
`pyl 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 whicharesterile
`solutions or suspensions can be utilized by, for example,
`intramuscular,
`intraperitoneal or subcutaneous injec-
`tion. Sterile solutions can also be administered intrave-
`nously. The compound can also be administered orally
`either in liquid or solid composition form.
`Preferably, the pharmaceutical compositionis 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 exam-
`ple, packeted powders, vials, ampoules, prefilled syrin-
`ges or sachets containing liquids. The unit dosage form
`can be, for example, a capsule ortabletitself, 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 physi-
`cian.
`
`In addition, the compounds ofthis invention may be
`employedas a solution, cream, or lotion by formulation
`with pharmaceutically acceptable vehicles containing
`0.1-5 percent, preferably 2%, of active compound
`which maybe administered to a fungally affected area.
`The following examplesillustrate the preparation of
`representative compoundsofthis invention.
`EXAMPLE1
`
`Rapamycin, 42-ester with trifluoroacetic acid
`Method A
`
`To a solution of 274 mg (0.3 mmol) of rapamycin in
`10 ml of methylene chloride is added 63 mg (0.3 mmol,
`43 y)) oftrifluoroacetic anhydride and then 39 mg (0.3
`mmol) of diisopropylethylamine (Hunig’s base). The
`mixture is stirred for 30 min and then an additional 78
`mg (0.6 mmol, 106 pl) of Hunig’s base is added followed
`by 63 mg (43 yl, 0.3 mmol)oftrifluoroacetic anhydride.
`After 30 min the reaction mixture is diluted with
`ethyl ether and aqueous sodium bicarbonate. The mix-
`ture is extracted with ethyl ether (3 times) and the com-
`bined organic extracts are washed with aqueous sodium
`bicarbonate. The resulting extract is dried over magne-
`sium sulfate and removedofsolvent in vacuo to give a
`solid foam.
`Flash chromatography on silica gel using ethy!
`ether:hexane (4:1) as the eluant gave 96 mg of product
`which is recrystallized from cyclohexane:hexane (1:1)
`to give 68 mg ofthe title compound: m.p. 87°-91° C.
`Anal Caled. for Cs53H7gF3NOj}4: C, 63.02; H, 7.78; N,
`1.39. Found: C, 62.85; H, 7.84; N, 1.41.
`
`Method B
`
`A solution of 914 mg (1 mmol) of rapamycin, 423 mg
`(1 mmol) of CMC (l-cyclohexyl-3-(2-morpholinoe-
`thyl)carbodiimide metho-para-toluenesulphonate), 20
`
`ay 0
`
`20
`
`25
`
`45
`
`55
`
`65
`
`5,100,883
`
`6
`1
`mg of dimethylaminopyridine and 101 mg (140 yl,
`mmol) of triethylamine in 50 ml] of methylene chloride
`is treated dropwise with 114 mg (1 mmol, 77 pl) of
`trifluoroacetic acid. The mixture is maintained with
`stirring for 17 h. An additional 1 mmol of CMC,20 mg
`of dimethylaminopyridine, 1 mmolof triethylamine and
`1 mmol!of trifluoroacetic acid is added. Additional
`trifluoroacetic acid (1 mmol) is added at 0.5 h intervals
`until] conversion is complete.
`The mixtureis diluted with water and ethyl ether and
`is extracted with ethy! ether. The ethereal] extract is
`washed with water, 0.5N hydrochloric acid andfinally
`with saturated aqueous sodium bicarbonate. The solu-
`tion is dried over magnesium sulfate, filtered and evapo-
`rated to give 870 mg of a white foam. The crude prod-
`uct is subjected to flash chromatography using hex-
`ane:ethy] ether (1:4) as the eluant. The obtained product
`is recrystallized from hexane:cyclohexane (1:1) to af-
`ford 397 mg ofthe title compound: m.p. 88°-91° C.
`Anal. Calcd. for Cs3H7gF3NQO}4: C, 63.02; H, 7.78; N,
`1.39. Found: C, 63.21; H, 7.67; N, 1.34.
`The following representative compoundscan be pre-
`pared from rapamycin and the appropriate carboxylic
`acid by employing Method B used to preparethetitle
`compound in Example 1.
`Rapamycin, 42-ester with perfluorovaleric acid
`Rapamycin, 42-ester with perfluorodecanoic acid
`Rapamycin, 42-ester with perfluorooctanoic acid
`Rapamycin, 42-ester with 9-H perfluorononanoic acid
`EXAMPLE2
`
`Rapamycin, 42-ester with 2,3,3,3-tetrafluoropropanoic
`acid
`
`To a solution of 914 mg (1 mmol) of rapamycin in 20
`ml of methylene chloride is added 245 mg (1.1 mmol) of
`Ishikawa Reagent (N,N-diethyl 1,1,2,3,3,3-hexafluoro-
`propylamine). After 1 hr, an additional 84 mg (0.4
`mmol) of Ishikawa Reagent is added. The mixture is
`stirred for 2 h and then is diluted with ethyl ether and
`aqueous sodium bicarbonate. The ethyl ether layer is
`separated and the aqueous layer is reextracted with
`ethyl ether (2 times). The combined ethereal extracts
`are dried over magnesium sulfate, filtered and removed
`of solvent in vacuo to give 762 mg of a solid foam.
`A portion of the crude productis subjected tosilica
`gel chromatography using hexane:ethy! ether (1:4) as
`the eluant to give 310 mg of product (Rf=0.28 hex-
`ane:ethy] ether(1:4)). Recrystallization from hexane:cy-
`clohexane (3:1) gives, after high vacuum drying over
`phosphorus pentoxide, 246 mg of the title compound:
`m.p. 87°-90° C.;
`Anal. Calcd. for Cs4H79F4NO}4: C, 62.23; H, 7.64, N,
`1.34. Found: C, 62.97; H, 7.69; N, 1.07.
`
`EXAMPLE3
`
`Rapamycin, 42-ester with difluoroacetic acid
`A solution of 914 mg (1 mmol) of rapamycin, 846 mg
`(2 mmol) of CMC, 40 mg of dimethylaminopyridine and
`202 mg (280 pl, 2 mmol) of triethylamine in 50 ml of
`methylene chloride is treated dropwise with 192 mg
`(126 yl, 2 mmol) of difluoroacetic acid. The mixtureis
`maintained with stirring for 1 h and additional triethyl-
`amine and difluoroacetic acid is added until the conver-
`sion is complete.
`The mixtureis diluted with water and ethyl ether and
`is extracted with ethyl! ether. The ethereal extract is
`
`West-Ward Pharm.
`Exhibit 1043
`Page 004
`
`West-Ward Pharm.
`Exhibit 1043
`Page 004
`
`
`
`7
`washed with water, 0.5N hydrochloric acid and finally
`with saturated aqueous sodium bicarbonate. The solu-
`tion is dried over magnesium sulfate, filtered and evapo-
`rated to give a white foam. The crude productis crys-
`tallized from cyclohexane to give 703 mg product.
`Flash chromatography using hexane:ethyl ether(1:4) as
`the eluant followed by recrystallization from cyclohex-
`ane affords the title compound: m.p. 87°-89° C.
`Anal. Calcd. for Cs3H79F2NO}a: C, 64.16; H, 8.03; N,
`1.41. Found: C, 64.46; H, 8.05; N, 1.42.
`EXAMPLE4
`
`Rapamycin, 42-ester with pentafluoropropionic acid
`A solution of 914 mg (1 mmol) of rapamycin, 846 mg
`(2 mmol) of CMC,40 mg of dimethylaminopyridine and
`202 mg (280 yl, 2 mmol) of triethylamine in 50 ml of
`methylene chloride is treated dropwise with 328 mg
`(210 pl, 2 mmol) of pentafluoropropionic acid. The
`mixture is maintained with stirring for 1 h and addi-
`tional
`triethylamine and difluoroacetic acid is added
`until the conversion is complete.
`The mixture is diluted with water and ethyl ether and
`is extracted with ethyl ether. The ethereal extract is
`washed with water, 0.5N hydrochloric acid and finally
`with saturated aqueous sodium bicarbonate. The solu-
`tion is dried over magnesium sulfate, filtered and evapo-
`rated to give a white foam. The crude productis crys-
`tallized from cyclohexane to give 703 mg product.
`Flash chromatography using hexane:ethyl ether (1:4) as
`the eluant followed by recrystallization from hexane:cy-
`clohexane(1:1) affords the title compound: m.p. 90°-97°
`Cc.
`
`Anal. Calcd. for CsgH7gFsNOya: C, 61.18; H, 7.42; N,
`1.32. Found: C, 61.45; H, 7.42; N, 1.33.
`Whatis claimedis:
`1. A compoundofthe formula
`
`OR!
`
` Ox
`
`HO
`
`wherein
`R! and R? are each, independently, hydrogen or
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`45
`
`50
`
`55
`
`1
`—crR3.
`
`R3 is a mono-, di-, poly-, or per-fluorinated alkyl
`group of 1-10 carbon atoms; with the proviso that
`R! and R? are both not hydrogen.
`2. A compoundof claim 1 where R?2is hydrogen.
`
`65
`
`5,100,883
`
`8
`3. A compound of claim 1 where R3is a mono-, di-,
`poly-, or per-fluorinated alkyl group of 1-6 carbon
`atoms.
`4. A compoundofclaim 1 where R? is hydrogen and
`R3is a mono-, di-, poly-, or per-fluorinated alkyl group
`of 1-6 carbon atoms.
`5. A compound of claim 1 which is rapamycin, 42-
`ester with trifluoroacetic acid.
`6. A compound of claim 1 which is rapamycin, 42-
`ester with 2,3,3,3-tetrafluoropropanoic acid.
`7. A compound of claim 1 which is rapamycin, 42-
`ester with difluoroacetic acid.
`8. A compound of claim 1 which is rapamycin, 42-
`ester with pentafluoropropionic acid.
`9. A method of treating transplantation rejection,
`host vs. draft disease, autoimmunediseases, and diseases
`ofinflammation in a mammal by administering an effec-
`tive amount of a compound having the formula
`
`OR!
`
`
`
`/
`wherein
`R! and R? are each, independently, hydrogen or
`
`OR!
`
`
`
`R3is a mono., di-, poly-, or per-fluorinated alkyl group
`of 1-10 carbon atoms; with the proviso that R! and R2
`are both not hydrogen.
`10. A pharmaceutical composition for use as an im-
`munosuppressive agent comprising an immunosuppres-
`sive amount of a compoundofclaim 1.
`11. A composition as claimed in claim 10 in unit dos-
`age form.
`*
`s
`2
`*
`*
`
`West-Ward Pharm.
`Exhibit 1043
`Page 005
`
`West-Ward Pharm.
`Exhibit 1043
`Page 005
`
`