United Stat~s Patent £191
`Caufield et al.
`
`[11] Patent Number:
`[45] Date of Patent:
`
`5,023,264
`Jun. 11, 1991
`
`[54] RAPAMYCIN OXIMES
`
`[75]
`
`Inventors: Craig E. Caufield, Plainsboro;
`Amedeo A. Failli, Princeton Juncton,
`both of N.J.
`
`[73] Assignee: American Home Products
`Corporation, New York, N.Y.
`
`[21] Appl. No.: 553,720
`
`[22] Filed:
`
`Jul. 16, 1990
`
`Int. Cl.' ................... A61K 31/395; C07D 491/16
`[51]
`[52] u.s. Cl ..................................... 514/291; 540/456;
`546/90
`[58] Field of Search .......................... 540/456; 546/90;
`514/291
`
`[56]
`
`References Cited
`U.S. PATENT DOCUMENTS
`3,929,992 12/1975 Sehgal ................................. 424/122
`3,993,749 11/1976 Sehgal ................................. 424/122
`4,316,885 2/1982 Rakhit ......................... : ....... 424/122
`4,401,653 4/1983 Eng ..................................... 424/114
`4,650,803 3/1987 Stella ................................... 514/291
`4,885,171 12/1989 Surendra ............................. 424/122
`OTHER PUBLICATIONS
`J. Antibiot. 28, 721-726 (1975).
`J. Antibiot. 28, 727-732 (1975).
`J. Antibiot. 31, 539-545 (1978).
`Can. J. Physiol. Pharmacal. 55, 48 (1977).
`Faseb 3, 3411, 5256 (1989).
`Lancet, 1183, (1978).
`J. Am. Chern. Soc. 103, 3215 (1981).
`Immunology, C. V. Moseby Co., pp. 12.8-12.11 (1989).
`
`Primary Examiner-Robert T. Bond
`Attorney, Agent, or Firm-Richard K. Jackson
`
`ABSTRACf
`[57]
`A derivative of rapamycin in which the 27-position has
`the structure
`
`wherein
`R 1 is hydrogen, alkyl, or -CH2Ar;
`Ar is
`
`wherein R2, R3, and R4 are each, independently,
`hydrogen, alkyl, aralkyl, alkoxy, hydroxy, cyano,
`halo, nitro, carbalkoxy, trifluoromethyl, amino, or
`a carboxylic acid;
`X is N, 0, or S;
`or a pharmaceutically acceptable salt thereof, which is
`by virtue of its immunosuppresive activity is useful in
`treating transplantation rejection host vs. graft disease,
`autoimmune diseases, and diseases of inflammation.
`
`9 Claims, 1 Drawing Sheet
`
`BRK-EVR-005961 0
`
`NOVARTIS EXHIBIT 2117
`Par v Novartis, IPR 2016-00084
`Page 1 of 6
`
`

`
`U.S. Patent
`
`June 11, 1991
`
`5,023,264
`
`RAPAMYCIN
`
`FIG. 1
`
`BRK-EVR-0059611
`
`NOVARTIS EXHIBIT 2117
`Par v Novartis, IPR 2016-00084
`Page 2 of 6
`
`

`
`5
`
`BACKGROUND OF THE INVENTION
`This invention relates to novel oxime derivatives of
`ra_pamycin and a method for using them in the treatment
`of transplantation rejection, host vs. graft disease, auto(cid:173)
`immune diseases, and diseases of inflammation.
`Rapamycin is a macrocyclic triene antibiotic pro- 10
`duced by Streptomyces hygroscopicus, which was found
`to have antifungal activity, particularly against Candida
`albicans, both in vitro and in vivo [C. Vezina et al., J.
`Antibiot. 28, 721 (1975); S. N. Seghal et al., J. Antibiot. 15
`28, 727 (1975); H. A. Baker et al., J. Antibiot. 31, 539
`(i978); U.S. Pat. No. 3,922,992; and U.S. Pat. No.
`3,993,749].
`Rapamycin alone (U.S. Pat. No. 4,885,171) or in com(cid:173)
`bination with picibanil (U.S. Pat. No. 4,401,653) has 20
`been shown to have antitumor activity. R. Martel et al.
`[Can. J. Physiol. Pharmacal. 55, 48 (1976)] disclosed
`that rapamycin is effective in the experimental allergic
`encephalomyelitis model, a model for multiple sclerosis; 25
`in the adjuvant arthritis model, a model for rheumatoid
`arthritis; and effectively inhibited the formation of 1GB(cid:173)
`like antibodies.
`The immunosuppressive effects of rapamycin have 30
`been disclosed in FASEB 3, 3411 (1989), rapamycin has
`been shown to be effective in inhibiting transplant rejec(cid:173)
`tion (U.S. patent application Ser. No. 362,544 filed Jun.
`6, 1989). Cyclosporin A and FK-506, other macrocyclic
`molecules, also have been shown to be effective as im- 35
`munosuppressive agents, therefore useful in preventing
`transplant rejection [FASEB 3, 3411 (1989); and R. Y.
`·
`Caine et al., Lancet 1183 (1978)].
`Mono- and diacylated derivatives of rapamycin (es- 40
`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 water soluble prodrugs of rapamycin
`(U.S. Pat. No. 4,650,803). Recently, the numbering
`convention for rapamycin has been changed; therefore 45
`according to Chemical Abstracts nomenclature, the
`esters described above would be at the 31- and 42-posi(cid:173)
`·tions.
`
`so
`
`DESCRIPTION OF THE INVENTION
`This invention provides oxime derivatives of rapamy(cid:173)
`cin which are useful as immunosuppressive and anti-in(cid:173)
`flammatory agents possessing the general structure of
`rapamycin shown in FIG. 1, where the 27 position has 55
`been transformed into an oxime having the structure
`
`;;{y•'.-zd}-.,,--f\"•',oc
`~R4 N
`'x\_
`
`R3
`
`wherein R2, R3, and R4 are each, independently,
`hydrogen, alkyl of 1-6 carbon atoms, aralkyl of
`7-10 carbon atoms, alkoxy of 1-6 carbon atoms,
`hydroxy, cyano, halo, nitro, carbalkoxy of 2-7
`carbon atoms, trifluoromethyl, amino, or a carbox(cid:173)
`ylic acid;
`X is N, 0, or S;
`or a pharmaceutically acceptable salt thereof.
`Of the compounds, preferred members are those in
`which Rl is hydrogen, alkyl of 1-6 carbon atoms, or
`
`The pharmaceutically acceptable salts may be formed
`from inorganic cations such as sOdium, potassium, and
`the like.
`Although compounds of this invention can be pre(cid:173)
`pared by conventional methods that are disclosed in the
`literature, because functional groups are contained in a
`large macrocyclic ring, functional group reactivity can(cid:173)
`not be readily predicted [R. B. Woodward et al., J. Am.
`Chem. Soc. 103, 3215 (1981)]. Rapamycin has carbonyl
`groups at the 14- and 27- and 33-positions. Based on
`x-ray crystallographic analysis, the 33-position was
`predicted to be the most reactive center; unexpectedly
`however, oxime formation occurred exclusively at the
`27-position.
`The compounds of the invention can be prepared ·by
`the following route from rapamycin.
`
`~
`II
`0
`
`1
`
`RAPAMYCIN OXIMES
`
`5,023,264
`
`2
`
`27
`
`y
`N
`I
`R10
`
`wherein
`Rl is hydrogen, alkyl of 1-6 carbon atoms, or -CH-
`2Ar;
`Ar is
`
`Immunosuppressive activity was evaluated in an in
`vitro standard pharmacological test procedure to mea(cid:173)
`sure lymphocyte proliferation (LAF) and in two in vivo
`60 standard pharmacological test procedures. The first in
`vivo procedure was a popliteal lymph node (PLN) test
`procedure which measured the effect of compounds of
`this invention on a mixed lymphocyte reaction and the
`second in vivo procedure evaluated the survival time of
`a pinch skin graft.
`The comitogen-induced thymocyte proliferation pro(cid:173)
`cedure (LAP) was used as an in vitro measure of the
`immunosuppressive effects of representative com-
`
`65
`
`BRK-EVR-0059612
`
`NOVARTIS EXHIBIT 2117
`Par v Novartis, IPR 2016-00084
`Page 3 of 6
`
`

`
`3
`pounds. Briefly, cells from the thymus of normal
`BALB/c mice are cultured for 72 hours with PHA and
`IL-l and pulsed with tritiated thymidine during the last
`six hours. Cells are cultured with and without various
`concentrations of rapamycin, cyclosporin A, or test 5
`compound. Cells are harvested and incorporated; radio(cid:173)
`activity is determined. Inhibition of lymphoprolifera(cid:173)
`tion is assessed in percent change in counts per minute
`from non-drug treated controls. The results are ex(cid:173)
`pressed by the following ratio:
`
`5,023,264
`
`4
`TABLE 1-continued
`PLN•
`LAP*
`(ratio)
`(ratio)
`+
`1.0
`
`Skin Graft
`(days+ SD)
`9.5 ± 1.4
`12.0 ± 1.7
`
`Compound
`
`0.007
`Example 3
`1.0
`Rapamycin
`*Calculation of ratios was described supra.
`+ Not evaluated
`
`3H-control thymus cells H3-rapamycin-treated thymus cells
`3H-control thymus cells - H3-test compound-treated cells
`A mixed lymphocyte reaction (MLR) occurs when 15
`lymphoid cells from genetically distinct animals are
`combined in tissue culture. Each stimulates the other to
`undergo blast transformation which results in increased
`DNA synthesis that can be quantified by the incorpora- 20
`tion of tritiated thymidine. Since stimulating a MLR is a
`function of disparity at Major Histocompatibility anti(cid:173)
`gens, an in vivo popliteal lymph node (PLN) test proce(cid:173)
`dure closely correlates to host vs. graft disease. Briefly,
`irradiated spleen cells from BALB/c donors are in- 25
`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 Day 4, tritiated thymidine is given i.p.,
`b.i.d. On Day 5, the hind popliteal lymph nodes are
`removed and dissolved, and radioactivity counted. The 30
`corresponding left PLN serves as the control for the
`PLN from the injected hind foot. Percent suppression is
`calculated using the non-drug treated animals as allo(cid:173)
`genic control. Rapamycin at a dose of 6 mg!kg, p.o.
`gave 86% suppression, whereas cyclosporin A at the 35
`same dose gave 43% suppression. Results are expressed
`by the following ratio:
`
`3H-PLN cells control C3H mouse -
`3H-PLN cells rapamycin-treated C3H mouse
`3H-PLN cells control C3H mouse -
`3H-PLN cells test compound-treated C3H mouse
`
`The results of these standard pharmacological test
`10 procedures demonstrate immunosuppressive activity
`both in vitro and in vivo for the compounds of this
`invention. Positive ratios in the LAF and PLN test
`procedures indicate suppression ofT cell proliferation.
`Transplanted pinch skin grafts are typically rejected
`within 6-7 days without the use of an immunosuppres(cid:173)
`sive agent. The increased survival time of the skin graft
`when treated with the compounds of this invention
`further demonstrates their utility as immunosuppressive
`agents. While it appears that the compound disclosed by
`Example 1 may cause T cell proliferation in the PLN
`test procedure, it is believed a negative ratio in this test
`procedure coupled with an increased survival time ob(cid:173)
`served in the skin graft test procedure indicates a prolif(cid:173)
`eration of Tsuppressor cells, which are implicated in sup(cid:173)
`pressing the immune response. (see, I. Roitt et al. Immu-
`nology, C. V. Moseby Co. 1989, p 12.8-12.11).
`Antifungal activity of the compounds of this inven(cid:173)
`tion was measured against 5 strains of Candida a/bicans
`using a plate test procedure for measurement of inhibi(cid:173)
`tion. The following represents the typical procedure
`used. Compound to be tested was placed on sterile dried
`1" plate disks, and allowed to dry. Agar plates were
`seeded with fungi and allowed to solidify. The impreg(cid:173)
`nated disks were placed on the seeded Agar surface and
`incubated for the time required for the particular cul-
`ture. Results are expressed in MIC (fLg/ml) to inhibit
`growth. Surprisingly, the oxime derivatives of this in(cid:173)
`vention have substantially less antifungal activity than
`40 the parent compound, rapamycin.
`TABLE2*
`Strain of Candida a/bicans
`
`•expressed as MIC (}.Lg/ml)
`
`The second in vivo test procedure is designed to
`A TCC A TCC A TCC
`10231
`38246
`38247 ATCC 38248
`Compound
`3669
`determine the survival time of pinch skin graft from 45 -~.......;.:,__.......;..;.,;.__;.,;..:..;.:,_...;..;.:,__.;;.;..;..:..:..:.:.:.;.:..._..;;.;;.;;,;__
`o.os
`male DBA/2 donors transplanted to male BALB/c
`Example I
`0.05
`0.2
`0.1
`0.2
`> 0·4
`Example 2
`> 0·4
`recipients. The method is adapted from Billingham R.
`>0.4
`>0.4
`>0.4
`E
`0.2
`Example 3
`0.2
`0.05
`0.05
`0.2
`. and Medawar P. B., J. Exp. Bioi. 28:385-(402), 1951.
`Rapamycin
`0.003
`O.D25
`0.003
`0.006
`O.D25
`Briefly, a pinch skin graft from the donor is grafted on
`the dorsum of the recipient as a homograft, and an 50
`autograft is used as control in the same region. The
`recipients are treated with either varying concentra(cid:173)
`tions of cyclosporin A as test control or the test com(cid:173)
`pound, intraperitoneally. Untreated recipients serve as
`rejection control. The graft is monitored daily and ob- 55
`servations are recorded until the graft becomes dry and
`forms a blackened scab. This is considered as the rejec(cid:173)
`tion day. The mean graft survival time (number of
`days±S.D.) of the drug treatment group is compared
`with the control group.
`The following table summarizes the results of repre(cid:173)
`sentative compounds of this invention in these three
`standard test procedures.
`TABLE 1
`PLN*
`LAP*
`(ratio)
`(ratio)
`0.25
`0.87
`
`Based on the results of these standard pharmacologi(cid:173)
`cal test procedures, the compounds are useful in the
`treatment of transplantation rejection such as, heart,
`kidney, liver, bone marrow, and skin transplants; auto(cid:173)
`immune diseases such as, lupus, rheumatoid arthritis,
`diabetes mellitus, myasthenia gravis, and multiple scle(cid:173)
`rosis; and diseases of inflammation such as, psoriasis,
`dermatitis, eczema, seborrhea, and inflammatory bowel
`disease.
`The compounds may be administered neat or with a
`pharmaceutical carrier to a mammal in need thereof.
`The pharmaceutical carrier may be solid or liquid.
`A solid carrier can include one or more substances
`which may also act as flavoring agents, lubricants, solu(cid:173)
`bilizers, suspending agents, fillers, glidants, compression
`aids, binders or tablet-disintegrating agents; it can also
`be an encapsulating material. In powders, the carrier is
`a finely divided solid which is in admixture with the
`
`60
`
`65
`
`Compound
`Example I
`Example 2
`
`Skin Graft
`(days+ SD)
`10.0 ± 1.6
`9.3 ± 1.4
`
`-1.40
`1.39
`
`BRK-EVR-0059613
`
`NOVARTIS EXHIBIT 2117
`Par v Novartis, IPR 2016-00084
`Page 4 of 6
`
`

`
`10
`
`40
`
`5,023,264
`
`6
`in hot ethyl acetate and upon cooling, white crystals
`formed. Vacuum filtration gave 23 mg (45%) of
`isomerically pure mono-oxime, which was biologically
`inac;:tive (results not presented).
`1H NMR (CDC}J, 400 MHz) 8 8.015 (s, 1H, NOH),
`4.820 (s, lH, COH), 3.414 (s, 3H, CH30-), 3.308 (s, 3H,
`CH30-), 1.667 (s, 3H, CH3C=C), 1.652 (s, 3H,
`CH3C=C); l3C NMR (CDCh, 100 MHz) 214.09
`(C=O), 192.03 (C=O), 169.38 (C=O), 167.21 (C=O),
`158.74 (C=NOH); IR (KBr) 3450 (OH), 2960 (CH),
`2890 (CH), 1760 (C=O), 1740 (C=O), 1732 (C=O),
`1660 (C=O, C=NOH), 1465, 1390, 1205, 1100, 1005
`cm-1; MS (neg. ion FAB) 928 (M-), 896,590,546, 167
`(100); High Res. MS (neg. ion FAB) Calc. for
`Cs1HsoN2013 928.5660, Found 928,5677.
`Analysis Calcd for Cs1HsoN2013.2 H20: C63.42; H
`8.21; N 2.74. Found: C63.47; H 8.11; N 2.60.
`The filtrate obtained following the crystallization
`described above was concentrated to give a white
`foamy solid. The solid was dissolved in hot ethyl ace(cid:173)
`tate and hexane was added until slightly cloudy. Upon
`cooling, white crystals formed. Vacuum filtration gave
`13 mg (25%) of pure mono oxime which was a geomet(cid:173)
`ric isomer of the compound isolated in the previous
`example. This isomer was tested and found to be active
`in the assays described supra.
`1H NMR (CDC}J, 400 MHz) 8 7.620 (s; lH, NOH),
`5.028 (s, lH, COH), 3.409 (s, 3H, CH30-), 3.306 (s, 3H,
`CH30-), 3.172 (s, 3H, CH30-), 1.807 (s, 3H,
`CH3C=C), 1.660 (s, 3H, CH3C=C); 13C NMR
`(CDC}J, 75 MHz) 211.83 (C=O), 191.43 (C=O), 168.73
`(C=O), 167.11 (C=O), 159.94 (C=NOH); IR (KBr)
`3450 (OH), 2960 (CH), 2920 (CH), 1775. (C=O), 1745
`(C=O), 1680, 1650 (C=O, C=NOH), 1475, 1400, 1220,
`1120, 1015 cm-1; MS (neg. ion FAB) 928 (M-), 167
`(100). High Res. MS (neg ion FAB) Calc. for
`Cs1HsoN2013 928.5660, Found 928.5660.
`Analysis Calcd for' Cs1HsoN2013: C 65.93; H 8.68; N
`3.01. Found: C 66.19; H 8.93; N 2.88.
`
`5
`fmely divided active ~gredient. In tablets, the active
`ingredient is mixed with a carrier having the necessary
`compression properties in suitable proportions and
`compacted in the shape and size desired. The powders
`and tablets preferably contain up to 99% of the active 5
`ingredient. Suitable solid carriers include, for example,
`calcium phosphate, magnesium stearate, talc, sugars,
`lactose, dextrin, starch, gelatin, cellulose, methyl cellu(cid:173)
`lose, sodium carboxymethyl cellulose, polyvinylpyr(cid:173)
`rolidine, low melting waxes and ion exchange resins.
`Liquid carriers are used in preparing solutions, sus(cid:173)
`pensions, emulsions, syrups, elixirs and pressurized
`compositions. The active ingredient can be dissolved or
`suspended in a pharmaceutically acceptable liquid car(cid:173)
`rier such as water, an organic solvent, a mixture of both 15
`or pharmaceutically acceptable oils or fats. The liquid
`carrier can contain other suitable pharmaceutical addi(cid:173)
`tives such as solubilizers, emulsifiers, buffers, preserva(cid:173)
`tives, sweeteners, flavoring agents, suspending agents,
`thickening agents, colors, viscosity regulators, stabiliz- 20
`ers or osmo-regulators. Suitable examples of liquid car(cid:173)
`riers for oral and parenteral administration include
`water (partially containing additives as above, e.g. cel(cid:173)
`lulose derivatives, preferably sodium carboxymethyl
`cellulose solution), alcohols (including monohydric 25
`alcohols and poly hydric 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(cid:173)
`pyl myristate. Sterile liquid carriers are useful in sterile 30
`liquid form compositions for parenteral administration.
`The liquid carrier for pressurized compositions can be
`halogenated hydrocarbon or other pharmaceutically
`acceptable propellent.
`Liquid pharmaceutical compositions which are sterile 35
`solutions or suspensions can be utilized by, for example,
`intramuscular, intraperitoneal or subcutaneous injec(cid:173)
`tion. Sterile solutions can also be administered intrave(cid:173)
`nously. The compound can also be administered orally
`either in liquid or solid composition form.
`Preferably, the pharmaceutical composition is in unit
`dosage form, e.g. as tablets or capsules. In such form,
`EXAMPLE2
`the composition is sun-divided in unit dose containing
`Rapamycin-0-benzyl-27-oxime
`appropriate quantities of the active ingredient; the unit
`dosage forms can be packaged compositions, for exam- 45
`To a solution of 50 mg (54. 7 tJ.mol) of rapamycin in 1
`ple, packeted powders, vials, ampoules, prefilled syrin-
`mL of methanol was added 12 mg (143 tJ.mol) of anhy-
`drous sodium acetate and 23 mg (143 tJ.mol) of ben-
`ges or sachets containing liquids. The unit dosage form
`zyloxyamine hydrochloride at room temperature. After
`can be, for example, a capsule or tablet itself, or it can be
`the appropriate number of any such compositions in
`72 h stirring at room temperature, the reaction was
`package form. The dosage to be used in the treatment 50 diluted with water and extracted three times with ethyl
`must be subjectively determined by the attending physi-
`acetate. The combined organics were washed with
`cian.
`brine, dried over anhydrous sodium sulfate, decanted,
`The following examples illustrate the preparation of
`and concentrated in vacuo to give a colorless oil. 1H
`representative compounds of this invention.
`NMR indicated a mixture of E and Z isomers. The oil
`EXAMPLE 1
`55 was dissolved in hot diisopropyl ether/hexane and upon
`cooling, white crystals formed. Vacuum filtration gave
`25 mg (45%) of pure mono-oxime.
`Rapamycin-27-oxime
`1H NMR (CDC}J, 400 MHz) 5.031 (s, 2H, CH2Ph),
`To a solution of 50 mg (54.7 tJ.mo1) ofrapamycin in 1
`3.365 (s, 3H, CH30-), 3.273 (s, 3H, CH30-), 3.118 (s,
`mL of methanol was added at room temperature, 12 mg
`(143 tJ.mol) of anhydrous sodium acetate and 10 mg (143 60 3H, CH30-), 1.661 (s, 3H, CH3C=C), 1.544 (s, 3H,
`CH3C=C); 13C NMR (CDC}J, 100 MHz) 211.65
`tJ.mol) of hydroxylamine hydrochloride. After 2 h stir-
`ring at room temperature, the reaction was complete by
`(C=O), 191.76 (C=O), 168.78 (C=O), 166.94 (C=O),
`TLC. The reaction was diluted with water and ex-
`158.46 (C=NOH); IR (KBr) 3450 (OH), 2940 (CH),
`tracted three times with ethyl acetate. The combined
`2890 (CH), 1750 (C=O), 1730 (C=O), 1650, 1630
`organics were washed with brine and dried over anhy- 65 (C=O, C=NOR), 1460, 1380, 1195, 1090, 990 cm- 1;
`drous sodium sulfate, decanted, and concentrated in MS (neg. ion FAB) 1018 (M-), 590, 546, 167 (100);
`vacuo to give a white, foamy solid. 1H NMR indicated
`High Res. MS (neg. ion FAB) Calc. for CssHs6N2013
`a mixture of E and Z isomers. The solid was dissolved
`1018.6130, Found 1018.6157.
`
`BRK-EVR-0059614
`
`NOVARTIS EXHIBIT 2117
`Par v Novartis, IPR 2016-00084
`Page 5 of 6
`
`

`
`7
`Analysis Calcd for CssHs6N20n.H20: C 67.18; H
`8.49; N 2.70. Found: C 67.17; H 8.61; N 2.56.
`
`5,023,264
`
`8
`
`X is N, 0, or S;
`or a pharmaceutically acceptable salt thereof.
`2. A compound of claim 1 wherein R' is alkyl of 1-6
`carbon atoms or a pharmaceutically acceptable salt
`5 thereof.
`3. A compound of claim 1 wherein R I is
`
`EXAMPLE3
`Rapamycin-0-methyl-27-oxime
`To a solution of 750 mg (820 J.Lmol) of rapamycin in
`15 mL of methanol was added 180 mg (2.15 mmol) of
`anhydrous sodium acetate and 180 mg (2.15 mmol) of
`methoxyamine hydrochloride at room temperature. 10
`After stirring overnight at room temperature, the reac(cid:173)
`tion mixture was diluted with water and extracted three
`times with ethyl acetate. The combined organics were
`washed with brine, dried over anhydrous sodium sul-
`fate, decanted and concentrated in vacuo to give a vis- 15 or a pharmaceutically acceptable salt thereof.
`cous oil. IH NMR indicated a mixture of E and z iso-
`4. A compound of claim 1 which is rapamycin-27-
`mers. The solid was dissolved in hot diisopropyl ether/-
`oxime or a pharmaceutically acceptable salt thereof.
`hexane and upon cooling, white crystals formed. Vac-
`5. A compound of claim 1 which is rapamycin-0-ben-
`uum filtration gave 370 mg (48%) of pure mono-oxime.
`zyl-27-oxime or a pharmaceutically acceptable salt
`IH NMR (CDCh, 400 MHz) 8 4.960 (bs, 1H, COH), 20 thereof.
`.
`.
`3.794 (s, 3H, CH30N=C), 3.395 (s, 3H, CH3o....:..), 3.288
`6. A compound of claun 1 which is rapamycin-0-
`(s, 3H, CHJO-), 3.121 (s, 3H, CHJO-), 1.645 (s, 3H,
`methyl-27-oxime or a pharmaceutically acceptable salt
`CH3C=C) 1.587 (s 3H CHJC=C)· 13C NMR
`thereof.
`(CDCb, 100 MHz) i11.10 (C=O), 192.11 (C=O),
`7. A metho? of treati~g transp~antation rejt;:ction,
`168.75 (C=O), 166.84 (C=O), 158.08 (C=NOMe); IR 25 ho~t vs. graft. d1s7ase, autotmmune d1s.e~es, ~nd diseases
`~fmflammat10n m a mammal by ~dm~1stenn~ an.effec-
`(KBr) 3450 (OH), 2940 (CH), 2890 (CH), 1750 (C=O),
`t1ve am~un~ of~ compound ~~1ch IS a denvatlve of
`1655, 1635 (C=O, C=NOCHJ), 1455, 1380, 1195, 1090,
`1050, 990 cm-1; MS (neg. ion FAB) 942 (M-), 590,
`rapamycm m whtch the 27-posttton has the structure;
`546, 167 (100); High Res. MS (neg. ion FAB) Calc. for
`Cs2Hs2N20n, 942.5818, Found 942.5863.
`Analysis Calcd for Cs2Hs2N20n.H20: C 65.00; H
`8.75; N 2.91. Found: C 65.20; H 8.83; N 2.50.
`What is claimed is:
`1. A derivative ofrapamycin in which the 27-position
`has the structure
`
`30
`
`35
`
`wherein
`Rl is hydrogen, alkyl of 1-6 carbon atoms, or -CH-
`2Ar;
`Ar is
`
`wherein
`R' is hydrogen, alkyl of 1-6 carbon atoms, or -CH-
`2Ar;
`Ar is
`
`40
`
`45
`
`50
`
`55
`
`wherein R2, R3, and R4 are each, independently,
`hydrogen, alkyl of 1-6 carbon atoms, aralkyl of
`7-10 carbon atoms, alkoxy of 1-6 carbon atoms,
`hydroxy, cyano, halo, nitro, carbalkoxy of 2-7 65
`carbon atoms, trifluoromethyl, amino, or a carbox(cid:173)
`ylic acid;
`
`wherein R2, R3, and R4 are each, independently,
`hydrogen, alkyl of 1-6 carbon atoms, aralkyl of
`7-10 carbon atoms, alkoxy of 1-6 carbon atoms,
`·hydroxy, cyano, halo, nitro, carbalkoxy of 2-7
`carbon atoms, trifluoromethyl, amino, or a carbox(cid:173)
`ylic acid;
`X is N, 0, or S;
`or a pharmaceutically acceptable salt thereof.
`8. A pharmaceutical composition comprising a com(cid:173)
`pound of claim 1 or a pharmaceutically acceptable salt
`thereof and a pharmaceutically acceptable carrier.
`9. A composition as claimed in claim 8, in unit dosage
`form.
`
`BRK-EVR-0059615
`
`NOVARTIS EXHIBIT 2117
`Par v Novartis, IPR 2016-00084
`Page 6 of 6