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  • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
  • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
  • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
• • 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
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • C—CHEMISTRY; METALLURGY
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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
  • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
  • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
  • C07D231/14—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
  • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
  • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
  • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
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  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
  • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
  • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
  • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
  • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
• • C—CHEMISTRY; METALLURGY
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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
  • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
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  • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
  • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
  • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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  • C07D—HETEROCYCLIC COMPOUNDS
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  • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
  • C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
  • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
  • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
  • C07D495/04—Ortho-condensed systems

Definitions

• T-cells play an important role in regulating immune response (F. Powrie and R.L. Coffman, Immunol. Today. 14, p. 270 (1993)). Indeed, activation of T- cells is often the initiating event in many inflammatory and autoimmune diseases.
• IL-2 is an autocrine growth factor which plays an essential role in the regulation of T-cell activation and proliferation. Clinical studies have shown that interference with IL-2 activity effectively suppresses immune response in vivo (T.A. Waldmann, Immunol. Today. 14, 270 (1993)). Accordingly, agents which inhibit IL-2 production are therapeutically useful for selectively suppressing immune response in a patient in need of such immunosuppression.
• N-[4-(3,5-dimethylpyrazol-l-yl)phenyl]acetam ⁇ de (Bouchet and Coquelet. Bull. Soc. Chim. Fr. 1976, 195), N-[4-(3-methyl-5-chloropyrazol-l-yl)phenyi]acetamide (Michaelis and Behn, Chem. Ber. 1900, 33, 2602), N-[4-(3-methyl-5-(methylth ⁇ o)pyrazol-l-yl)phenyl]acetamide (Michaelis, Justus Liebigs Ann.
• Matsuo (EP 418845 Al ) are l-[4-(C,-C 6 alkylamino)phenyl]- and [4-(C,-C 6 acylamino)phenyl]pyrazoles substituted on the pyrazole on either the 3-, 4-, or 5-pos ⁇ t ⁇ on with CF 3 , halogen, dimethvlammomethvl, CN. C, 6 aikylthio, or este ⁇ fied carboxy and on another of the 3-, 4-, or 5-pos ⁇ t ⁇ ons with a sunstituted aryl or heteroaryl ⁇ ng. No mention is made of inhibition of IL-2 production
• the compounds of this invention are 1 -(4-am ⁇ nophenyl)pyrazoles optionally substituted on the 3- and 5-pos ⁇ t ⁇ ons of the pyrazole ⁇ ng and on the ammo group on the 4-pos ⁇ t ⁇ on of the phenyl ⁇ ng having antiinflammatory activity by virtue of their ability to inhibit IL-2 production m T-lymphocytes
• the invention comp ⁇ ses l-(4-am ⁇ nophenyl)pyrazoles of
• R, and R- are the same or different ana each is CF 3 , halogen, CN, C, 8 alkyl or branched alkyl or C, g alkenyl or branched alkenyl or C 3 8 cycloalkyl optionally substituted with OH, CN or methoxy, C, 8 alkoxy, C alkyloxyalkyl, C, g aikylthio.
• R 5 is C, 4 alkyl or C alkenyl optionally substituted with carbocyclyl or heterocyclyl, aryl or heterocyclyl connected to the pyrazole in any position that makes a stable bond, optionally substituted with halogen, C, 4 alkyl, C alkenyl, CN, Me : N. CO 2 Me, OMe, aryl, heterocyclyl or R s
• R 2 is H, halogen, or methyl L is -NHC(O)-, -NHC(0)0-, -NHC(0)C(0)-, -NHC(S)-, -NH-, -NHC(O)NH, NHC(S)NH, NHCH 2 , -NHCH(R «)- , where R 6 is H, CN, C, 6 alkvl, C, 6 alkyloxyoalkyl C, 6 alkythioalkyl, C 1 6 alkylsulfinylalkyl, C, 6 alkysulfonylalkyl, C 3 6 cycloalkyl, or heterocyclyl or aryl optionally substituted with a halogen, C, 4 alkyl, CN, Me,N, CO 2 Me or OMe, or -NHC(R 6 )- lower alkyl.
• R 4 is C 1 8 alkyl, C, 8 alkyloxy, C, 8 aikylthio, C, 8 alkylammo, C, 4 alkoxyalkyl, C alkylthioalkyl, C, 4 alkylaminoalkyl C dialkylalkylaminoalkyl, carbocyclyl or heterocyclyl, optionally substituted with one or more halogen, -CN, -NO 2 , SO 2 NH 2 , or R 7 where R 7 is phenyl, heterocyclyl, C 3 6 cycloalkyl, C, 6 alkyl, C 2 6 alkenyl, C, 6 alkyloxyalkyl, C, 6 alkylthioalkyl, C 1 6 alkylsulfinylalkyl, C 1 6 alkylsulfonylalkyl or C 2 6 alkynyl, optionally substituted with halogen, OH, alkyloxy, CN, COO-lower al
• phenyl or heterocylcyl C0 2 R, -N(R 7 ) 2 , -NH(R 7 ), -C(O)R 7 , -OR 7 , S(0) n R 7 where n is 0, 1 or 2, -SO : NHR 7 , -SO,N(R 7 ) 2
• BOC or t-BOC is tertiary butoxycarbonyl
• DMAP is 4-d ⁇ methylammo py ⁇ dme
• DIBAL is dnsobutylalummum hyd ⁇ de
• Oxz is oxazole
• Ph is phenyl
• Pr is propyl
• PyBOP is Benzotnazole-1-yl-oxy-t ⁇ s-pyrrohd ⁇ no-phosphon ⁇ um hexafluorophosphate
• Th is thiophene
• THF is tetrahydrofuran
• Thz is thiazole
• Rt room temperature
• EDC is l-(3-dimethylaminopropyl)-3-ethylcarbodimide hydrochloride.
• alkyl refers to a saturated aliphatic radical containing from one to ten carbon atoms.
• Alkyl refers to both branched and unbranched alkyl groups. Preferred alkyl groups are straight chain alkyl groups containing from one to eight carbon atoms and branched alkyl groups containing from three to eight carbon atoms. More preferred alkyl groups are straight chain alkyl groups containing from one to six carbon atoms and branched alkyl groups containing from three to six carbon atoms.
• Alkyl includes unsubstituted alkyl radicals, those radicals that are partially or fully halogenated and those radicals substituted with one to four, preferably one or two, substituents selected from amino, cyano, nitro, methoxy, ethoxy and hydroxy.
• cycloalkyl refers to the cyclic analog of an alkyl group, as defined above.
• Preferred cycloalkyl groups are saturated cycloalkyl groups containing from three to eight carbon atoms, and more preferably three to six carbon atoms.
• Alkyl and “cycloalkyl”, as used herein, include unsubstituted alkyl and cycloalkyl radicals.
• alkenyl and alkynyl refer to a mono- or polyunsaturated aliphatic hydrocarbon radical containing from two to twelve carbon atoms, containing at least one double or triple bond, respectively.
• alkenyl and alkynyl refer to both ranched and unbranched alkenyl and alkynyl groups.
• Preferred alkenyl and alkynyl groups are straight chain alkenyl or alkynyl groups containing from two to eight carbon atoms and branched alkenyl or alkynyl groups containing from five to ten carbon atoms.
• More prefe ⁇ ed alkenyl and alkynyl groups are straight chain alkenyl or alkynyl groups containing from two to six carbon atoms and branched alkenyl or alkynyl groups containing from five to eight carbon atoms.
• cycloalkenyl refers to the cyclic analog of an alkenyl group, as defined above.
• Preferred cycloalkenyls include cycloalkenyl rings containing from three to eight carbon atoms, and more preferably, from three to six carbon atoms.
• alkenyl include unsubstituted alkenyl or alkynyl radicals, those radicals that are partially or fully halogenated and those radicals substituted with one to four, preferably one or two, substituents selected from halo, ammo, cyano, nitro, methoxy, ethoxy and hydroxy.
• aryl refers to phenyl and naphthyl, phenyl and naphthyl that are partially or fully halogenated and phenyl and naphthyl substituted with halo, alkyl, hydroxyl, nitro, -COOH. -CO(lower alkoxy), -CO(lower alkyl), ammo, alkylamino, dialkylamino, alkoxy, - NCOH, -NCO(lower alkyl), -NSO,-Ph(halo) 0 . 3 , Ph.
• carboxy alkyl refers to an alkyl radical containing a -COOH substituent.
• halo refers to a halogen radical selected from fluoro, chloro, bromo or iodo. Preferred halo groups are fluoro. chloro and bromo.
• ''carbocyclyl refers to a stable 3-8 membered (but preferably, 5 or 6 membered) monocyclic or 7-11 membered bicyclic radical which may be either saturated or unsaturated, aromatic or non-aromatic.
• Preferred carbocycles include, for example, phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, indanyl, indenyl, dihydronaphthyl and tetrahydronaphthyl.
• Carbocyclyl refers to unsubstituted carbocyclic radicals, those radicals that are partially or fully halogenated and those radicals substituted with alkyl; hydroxyl; nitro; - COOH; -CO(lower alkoxy); -CO(lower alkyl); ammo; alkylamino; dialkylamino; alkoxy; - NCHO; -NCO(lower alkyl); -NSO : -Ph(halo) 0 3 , Ph: -O-Ph; naphthyl; -O-naphthyl; pyrrolyl; pyrrolyl substituted with lower alkyl; py ⁇ dyl; pyndmyl, pyrazinyl; py ⁇ midmyl
• heterocycle refers to a stable 5-8 membered (but preferably, 5 or 6 membered) monocyc c or 8-11 membered bicyc c heterocycle radical which may be either saturated or unsaturated, aromatic or non-aromatic, and which may be optionally benzo- or pyridofused if monocychc.
• Each heterocycle consists of carbon atoms and from 1 to 4 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur.
• nitrogen and “sulfur” include any oxidized form of nitrogen and sulfur and the quatermzed form of any basic mtrogen.
• the heterocycle may be attached by any atom of the cycle, which results m the creation of a stable structure.
• Preferred heterocycles include, for example, benzimidazolyl, furyl; lmidazolyl, lmidazohnyl, lmidazohdinyl, qumohnyl, isoqumolinyl, indolyl, oxazolyl, py ⁇ dyl, pyrrolyl, pyrrohnyl, pyrazolyl, pyrazinyl, py ⁇ midinyl, py ⁇ dazinyl, quinoxolyl, prperidmyl, morphohnyl, thiomorphohnyl, furyl, thienyl, t ⁇ azolyl, thiazolyl, ⁇ - carbolinyl, tetrazolyl, thiazolidinyl, benzofuranoyl, thiamorpholinyl sulfone, benzoxazolyl, oxopipe ⁇ dinyl, oxopyrroldinyl, oxoazepmy
• heterocycles of this invention include lmidazolyl, py ⁇ dyl, pyrrolyl, pyrazolyl, piperidinyl, morpholinyl, furyl, thienyl, thiazolyl and the benzo- and py ⁇ do-fused de ⁇ vatives thereof.
• Heterocyclyl refers to unsubstituted heterocycle radicals, those radicals that are partially or fully halogenated and those radicals substituted with alkyl; hydroxyl; nitro; - COOH, -CO(lower alkoxy), -CO(lower alkyl); amino; alkylamino; dialkylamino, alkoxy; - NCHO; -NCO(lower alkyl); -NSO 2 -Ph(halo) 0 3 , Ph; -O-Ph; naphthyl; -O-naphthyl; pyrrolyl; pyrrolyl substituted with lower alkyl; py ⁇ dyl; py ⁇ dinyl; pyrazinyl; py ⁇ midinyl and pyridazinyl.
• lower used conjunction with other terms (e.g., "alkyl”, “alkoxy” and the like) refers to a radical containing from one to six, preferably from one to five and more preferably, from one to four carbon atoms.
• a “lower alkyl” group is a branched or unbranched alkyl radical containing from one to six carbon atoms.
• patient refers to a warm-blooded animal, and preferably a human.
• prevention or “prophylaxis” refers to a measurable reduction in the likelihood of a patient acquiring a disease or disorder.
• treatment refers to either the alleviation of the physical symptoms of a disease or an improvement in the physiological markers used to measure progression of a disease state.
• pharmaceutically acceptable carrier or “pharmaceutically acceptable adjuvant” refers to a non-toxic carrier or adjuvant that may be administered to a patient together with a compound of this invention and which does not destroy the pharmacological activity of that compound.
• pharmaceutically effective amount refers to an amount effective in suppressing the immunity of a patient in need of such treatment. Suppressed immunity can be readily measured by observing the degree of inhibition of IL-2 production in human T-cells (PBLs) by known techniques.
• prophylactically effective amount refers to an amount effective in preventing or reducing the likelihood of initial onset or progression of an immune disorder in a patient susceptible to such disorder.
• any compounds of this invention containing one or more asymmetric carbon atoms may occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. All such isomeric forms of these compounds are expressly included in the present invention.
• Each stereogenic center may be in the R or S configuration, or a combination of configurations.
• the compounds of this invention are defined to include pharmaceutically acceptable derivatives thereof.
• a "pharmaceutically acceptable derivative” refers to any pharmaceutically acceptable salt, ester, or salt of an ester of a compound of this invention, or any other compound which, upon administration to a patient, is capable of providing (directly or indirectly) a compound of this invention, a pharmacologically active metabolite or pharmacologically active residue thereof.
• the compounds of this invention may be used m the form of salts de ⁇ ved from morgamc or organic acids Included among such acid salts, for example, are the following- acetate, adipate, alginate. aspartate. benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate.
• glucoheptanoate glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochlo ⁇ de, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate.
• lactate maleate, methanesuifonate, 2-naphthalenesulfonate, mcotinate, oxalate, pectinate, persulfate, 3-phenylprop ⁇ onate, picrate, pivalate, propionate, succinate, tartrate, tosylate and undecanoate.
• This invention relates to substituted l-(4-ammophenyl)pyrazoles and analogs thereof that inhibit ⁇ nterieukm-2 (IL-2) production
• this invention relates to a novel class of substituted l-(4-ammophenyl)pyrazoles and pharmaceutical compositions compnsing these compounds
• This invention also relates to methods for producing such novel substituted 1 -(4-am ⁇ nophenyl)pyrazoles Because of their selective lmmunomodulating properties, the compounds and pharmaceutical compositions of this invention are particularly well suited for preventing and treating immune disorders, including autoimmune disease, inflammatory disease, organ transplant rejection and other disorders associated with IL-2 mediated immune response.
• the substituted l-(4-ammophenvi)pyrazoles of this invention are represented by Formula I
• R, and R 3 are the same or different and each is CF 3 , halogen, CN, C, 8 alkyl or branched alkyl or C, 8 alkenyl or branched alkenvl or C 3 8 cycloalkyl optionally substituted with OH, CN or methoxy, C, 3 alkoxy, C alkvloxyalkyl, C, 8 aikylthio, C alkylthioalkyl, C, 8 dialkylamino, C dialkylammoalkyl, CO : R 5 where R, is C, 4 alkyl or C, 4 alkenyl optionally substituted with carbocyclyl or heterocyclyl, aryl or heterocyclyl connected to the pyrazole in any position that makes a stable bond, optionally substituted with halogen, C, , alkyl, C alkenyl, CN, Me 2 N, CO,Me, OMe, aryl, heterocyclyl or R 5
• R 2 is H, halogen or methyl
• L is -NHC(O)-, - HC(0)0-, -NHC(0)C(0)-, -NHC(S)-, -NH-, -NHC(0)NH, NHC(S)NH, HCH Tavern -NHCH(R 6 )- , where R 6 is H, CN, C, 6 alkvl, C, 6 alkyloxyoalkyl C, 6 alkythioalkyl, C 1 6 alkylsulfinylalkyl, C, s alkysulfonylalkyl, C 3 6 cycloalkyl, or heterocyclyl or aryl optionally substituted with a halogen, C alkyl, CN, Me 2 N, C0 2 Me or OMe, or -NHC(R 6 )- lower alkyl
• R 4 is C, 8 alkyl, C, 8 alkyloxy, C, 8 aikylthio, C 1 8 alkylamino.
• novel substituted 1 -(4-aminophenyl)pyrazoles of Formula I are those wherein:
• R is straight-chained, branched or cyclo- C 3.8 alkyl, alkenyl, or alkynyl; C,. 3 alkyloxyalkyl, C,. 5 alkyloxy, C,. 3 alkylthioalkyl, C,. 5 aikylthio, CF 3 ; heterocyclyl or aryl optionally substituted with halogen, C alkyl, CN, alkoxy or Me 2 N;
• R is H
• R 3 is halogen, Me, Et, CF 3 , CN. cyclopropyl, vinyl, SMe, OMe, heterocyclyl or aryl optionally substituted with halogen, C alkyl, CN, alkoxy or Me,N;
• L is -NHC(O)-, -NH-, -NHC(0)NH, -C(0)NH, or -NHCH(R 6 )- ) where R 6 is H, C alkyl, or CN and
• R 4 is C ,. 6 alkyl, C,. 4 alkyloxyalkyl, C alkylthioalkyl, cyclohexyl, cyclopentyl, indanyl, indolyl, phenyl, thienyl, naphthyl, isoxazolyl or pyridyl, optionally substituted with one or more halogen, -CN, -N0 2 , S0 2 NH 2 . or R-, where R 7 C,. 6 alkyl, C 2 . 6 alkenyl, C,. 6 alkyloxyalkyl, C,.
• R is Et, z-Pr, t-Bu, cyclopentyl, CF 3 , -OEt, MeOCH 2 -, 2- or 3-tetrahydrofuranyl, 2-, 3-, or 4- pyridyl or 2-pyrazinyl;
• R 2 is H;
• R 3 is Halogen, CN, CF 3 , Me. 5Me or Et;
• L is -NHC(O)-, -NH- or -NHCH 2 -;
• R 4 is alkyl, cyclohexyl, cyclopentyl, indanyl, indolyl, phenyl, thienyl, naphthyl, or pyridyl, optionally substituted with one or more halogen, -CN, or R 7 where R 7 C,. 6 alkyl, C 2.6 alkenyl, C,. 6 alkyloxyalkyl, C,. 6 alkylthioalkyl, optionally substituted with OH, CN, or heterocylcyl; - CO 2 R 7, -N(R 7 ) 2 , -NH(R 7 ), -C(O)R 7 , or -OR 7 .
• a (4-aminophenyl)-3,5-disubstituted pyrazole 1 may be reacted with a carboxylic acid 2 under suitable coupling conditions known to one skilled in the art, for example EDC and a base catalyst such as N,N-dimethylaminopyridine in a suitable solvent such as methylene chloride acetonitrile or DMF (Method A).
• a suitable solvent such as methylene chloride acetonitrile or DMF
• 1 could be coupled with an acid halide 3 in the presence of a suitable base such as triethylamine in a suitable solvent such as methylene chloride (Method B).
• these compounds could be prepared by reacting a 4-(acetamido)phenylhydrazine 4 with a 1,5- disubstituted-2,4-pentanedione 5 in a suitable solvent such as acetic acid (Method C). If R, is different from R 3 , then two different products may form using Method C, which may be separated by techniques such as chromatography known to those skilled in the art.
• Compounds of Formula I in which L is -NH- and R 4 is a heteroaryl ring may be prepared, as illustrated below, by reaction of a (4-aminophenyl)-3,5-disubstituted pyrazole 1 with a heterocycle 6 containing a labile substituent such as a halogen, which may be displaced by nucleophilic substitution (Method D).
• the reaction may be carried out in a sealed tube or an open vessel, at ambient temperature or heated to 150°C in a suitable solvent such as dioxane or THF.
• a base such as sodium bis-trimethylsilyl amide may be added to the reaction.
• X is halogen or other leaving group
• L is -NHC(0)NH-
• a suitable solvent such as methylene chloride or toluene (Method E).
• An amine such as triethylamine may be added.
• 1 could be reacted with an amine carbonyl chloride such as N-morpholine carbonyl chloride 9 in a suitable solvent such as methylene chloride (Method F).
• the intermediate imine 12 reacted with a reducing agent such as sodium cyanoborohydride or sodium triacetoxyborohydride (Method I).
• a reducing agent such as sodium cyanoborohydride or sodium triacetoxyborohydride (Method I).
• 12 could be reacted with a nucleophile such as an alkyl or aryl lithium reagent (Method J).
• Pyrazole intermediates used in the preparations of the compounds of the invention may be prepared by methods known in the chemical literature. Two general methods that may be used are illustrated below. For example, a disubstituted 1 ,3-dione 13 may be heated with 4- nitrophenylhydrazine in a suitable solvent such as ethanol to provide a 3,5-disubstituted l-(4- nitrophenyl)pyrazole. If R, and R 3 are not equivalent, a mixture of two
• Method K a 3.5-disubstituted pyrazole may be reacted with nitrobenzene substituted in the 4-position with a leaving group such as a halogen in the presence of a base (Method L).
• a reducing agent such as SnCl 2 or hydrogen or a hydrogen source such as ammonium formate in the presence of a catalyst such as palladium.
• R, and R 3 can greatly enhance the regioselectivity of the reaction to produce 14 by Method L.
• R is aryl, and preferably an electron deficient aryl. such as pyridine, or a phenyl with an electron withdrawing group such as -CN or N0 2 .
• R is alkyl, preferably ethyl or methyl
• a leaving group preferably fluorine in a suitable solvent, preferably DMSO
• a suitable base preferably t-BuOK at about 75 °C for about one hour.
• R is alkyl in a regioselective manner using Method L, by employing a substituted pyrazole where R, contains a chelating group, preferably oxygen, and then converting R, in the resulting product 14 to an alkyl group.
• Method L by employing a substituted pyrazole where R, contains a chelating group, preferably oxygen, and then converting R, in the resulting product 14 to an alkyl group.
• R is an alkylcarbonyl, preferably - C(0)CH 3 , or an ether such as CH(R)OR', where R is preferably hydrogen or methyl, and R' is a suitable protecting group, preferably tetrahydropyranyl (THP) with a nitrobenzene substituted in the 4-position with a leaving group, preferably fluorine, in a suitable solvent, such as THF or DMSO.
• THF tetrahydropyranyl
• a suitable solvent such as THF or DMSO.
• a suitable base such as t-BuOK, n- BuLi, sodium hydride, or ethyl magnesium bromide, preferably t-BuOK
• R may be converted to an alkyl group as illustrated in Scheme II
• reaction with an acid such as p- toluenesulfonic acid in a suitable solvent such as aqueous methanol provides alcohol 14b.
• a suitable oxidizing agent such as pyridinium chlorochromate provides ketone 14c.
• Wittig reaction with a phosphorous ylide derived from methyltriphenylphosphonium bromide provides olefin 14d.
• Reaction under reducing conditions, such as hydrogenation in the presence of a palladium catalyst provides 15.
• R is an alkylcarbonyl, one would proceed beginning with 14c.
• 5-Dimethylaminomethylpyrazole 16 may be prepared by a general method (Method M) described in the chemical literature (Tang and Method M
• the 1 -(4-aminophenyl)pyrazole analog of 16 may then be prepared as described in Method L.
• nitrile 17 and 4- nitrophenylhydrazine Using nitrile 17 and 4- nitrophenylhydrazine. the 5 -amino and 5-disubstituted aminopyrazoles 18 and 19 can be prepared (Method N).
• Reaction of hydrazone 20, with acrylonitrile and iodobenzene diacetate, followed by oxidation of pyrazoline 21 provides 5-cyanopyrazoles 22 (Method O).
• the nitrophenylpyrazoles described in Methods N and O can then be reduced to the aminophenylpyrazoles as described in Method L.
• ketoester 23 Reaction of ketoester 23 with 4-nitrophenylhydrazine in a suitable solvent, preferably acetic acid at about reflux temperature provides 24.
• a halogenating agent such as phosphorous tribromide or phosphorous oxychloride
• Method R. 25a may be cross coupled with a terminal acetylene 26.
• W may be. for example, hydrogen, or an alkyl group or any other group not adversely affecting the reaction, using conditions described by T. Sakamoto et al., (Synthesis, 1983, 312) to provide an alkyne at R, (14a).
• Reaction with substituted or unsubstituted aryl- or heteroarylboronic acids under conditions described by N.
• Miyaura et al. (Chem. Rev. 1995, 95, 2457) provides 14c with aryl or heteroaryl groups at R 3 (Method T).
• Alkynes 14a and alkenes 14b may be converted to the corresponding alkyl groups by reduction with a suitable reducing agent such as hydrogen in the presence of a suitable catalyst such as platinum or palladium to provide 1 (see Method L), with an alkyl group at R 3 .
• a suitable reducing agent such as hydrogen
• a suitable catalyst such as platinum or palladium
• reaction with a reducing agent that leaves alkenes and alkynes intact, such as SnCl 2 provides 1 with alkenes or alkynes at R 3 .
• R 3 aryl or heteroaryl
• Method U describes an alternate procedure for preparing compounds of Formula I where L is -NH-.
• Intermediate 1 may be heated at about 70 °C with an aryl bromide in the presence of a palladium catalyst, preferably Pd 2 (dba) 3 , 2.2'-bis(diphenylphosphino)-l,l '-binapthyl (BINAP), and a base, preferably NaOt-Bu, in a solvent such as toluene, as described by S. Buchwald et al.(J. Amer. Chem. Soc. 1993, 119, 8451).
• a palladium catalyst preferably Pd 2 (dba) 3 , 2.2'-bis(diphenylphosphino)-l,l '-binapthyl (BINAP)
• BINAP 2.2'-bis(diphenylphosphino)-l,l '-binapthyl
• a base preferably NaOt-Bu
• the 1-phenylpyrazole analogs of Formula I inhibit production of IL-2.
• the compounds of this invention inhibit IL-2 production by T cells.
• This inhibition of IL-2 production is therapeutically useful for selectively suppressing immune function.
• the result of such selectively suppressed immunity includes reduced cell proliferation of peripheral blood lymphocytes and cellular immune response.
• the inhibition of IL-2 production is an attractive means for preventing and treating a variety of immune disorders, including inflammatory diseases, autoimmune diseases, organ and bone marrow transplant rejection and other disorders associated with IL-2 mediated immune response.
• the compounds of Formula I may be used to prevent or treat acute or chronic inflammation, allergies, contact dermatitis, psoriasis, rheumatoid arthritis, multiple sclerosis, type 1 diabetes, inflammatory bowel disease. Guillain-Barre syndrome. Crohn's disease, ulcerative colitis, graft versus host disease (and other forms of organ or bone marrow transplant rejection) and lupus erythematosus. Other disorders associated with IL-2 mediated immune response will be evident to those of ordinary skill in the art and can also be treated with the compounds and compositions of this invention.
• the compounds of this invention may be administered in any conventional dosage form in any conventional manner. Such methods of treatment, including their dosage levels and other requirements, may be selected by those of ordinary skill in the art from available methods and techniques.
• a compound of this invention may be combined with a pharmaceutically acceptable carrier or adjuvant for administration to a patient in need of such treatment in a pharmaceutically acceptable manner and in an amount effective to treat (including lessening the severity of symptoms) the immune disorder.
• the compounds of this invention may be administered alone or in combination with conventional therapeutics, such as conventional immunosuppressants.
• conventional therapeutics such as conventional immunosuppressants.
• combination therapies utilize lower dosages of the conventional therapeutics, thus avoiding possible toxicity and adverse side effects incurred when those agents are used as monotherapies.
• the compounds of this invention may be physically combined with the conventional therapeutics into a single pharmaceutical composition.
• the compounds may then be administered together in a single dosage form.
• the compounds may be administered separately (either serially or in parallel). Separate dosing allows for greater flexibility in the dosing regime.
• the compounds of Formula I and the pharmaceutical compositions containing those compounds may be administered to a patient in any conventional manner and in any pharmaceutically acceptable dosage from, including, but not limited to, intravenously, intramuscularly, subcutaneously, intrasynovially, by infusion, sublingually. transdermally. orally, topically or by inhalation.
• the preferred modes of administration are oral and intravenous.
• Dosage forms of the compounds of this invention include pharmaceutically acceptable carriers and adjuvants known to those of ordinary skill in the art. These carriers and adjuvants include, for example, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, buffer substances, water, salts or electrolytes and cellulose-based substances.
• Preferred dosage forms include, tablet, capsule, caplet. liquid, solution, suspension, emulsion, lozenges, syrup, reconstitutable powder, granule, suppository and transdermal patch. Methods for preparing such dosage forms are known (see, for example, H.C. Ansel and N.G. Popovish, Pharmaceutical Dosage Forms and Drug Delivery Systems, 5th ed..
• Dosage levels and requirements are well-recognized in the art and may be selected by those of ordinary skill in the art from available methods and techniques suitable for a particular patient. Typically, dosage levels range from about 10-1000 mg/dose for a 70 kg patient. Although one dose per day may be sufficient, up to 5 doses per day may be given. For oral doses, up to 5000 mg/day may be required. As the skilled artisan will appreciate, lower or higher doses may be required depending on particular factors. For instance, specific dosage and treatment regimens will depend on factors such as the patient's general health profile, the severity and course of the patient's disorder or disposition thereto and the judgment of the treating physician. Synthetic Examples
• Example 1 Synthesis of N- ⁇ 4-[3,5-bis(trifluoromethyl)pyrazol-l-yl]phenyl ⁇ cyclo- hexanecarboxamide (Method A).
• Triethylamine (0.05 mL. 0.36 mmol) was added to ⁇ -(+)-3-aminotetrahydrofuran toluene-4- sulfonate (Fluka. 0.082 g, 0.32 mmol) and 6-8 beads of 4A molecular sieves in an oven-dried sealed tube.
• l-(4'-Isocyanatophenyl)-3,5-di(trifluoromethyl)pyrazole (0.10 g, 0.32 mmol) in toluene (3 mL) was added, and the mixture was capped and stirred at room temperature for two and a half days. The solid product was collected by filtration, and washed with toluene.
• Example 8 Synthesis of morpholine-4-carboxylic acid ⁇ 4-[3.5- bis(trifluoromethyl)pyrazol-l-yl]phenyl ⁇ amide (Method F)
• Example 13 Synthesis of l-(4'-aminophenyl)-3-t-butyl-5-trifluoromethylpyrazole and 1- (4'-aminophenyl)-5-t-butyl-3-trifluoromethylpyrazole (Method K)
• N- Alkylated material 1.5 g (68%). was also obtained. 6-(3-cyanopropyloxy)n ⁇ cotimc acid, methyl ester (1.8 mmol) was dissolved in methanol/water (16 mL:4 mL) and treated with LiOH monohydrate (2.7 mmol). After sti ⁇ ing overnight, volatiles were removed and reaction neutralized with IN sulfuric acid. The solid product was collected by filtration, washed with water and dried to give 6-(3- cyanopropyloxy)nicotinic acid (315 mg. 85%).
• Example 28 Synthesis of 6-[(3-[l,3]dioxolan-2-yl-propoxy)]-N-[4-(5-cyano-3-pyridin-3- yl-pyrazol-l-yl)phenyl]nicotinamide
• the pyrazole from above (16.2 g, 61.3 mmol) was dissolved in 1 : 1 mixture of THF and CH 2 C1 2 (300 mL). and nicotinic acid (8.3 g, 67.4 mmol) and EDC (14.1 g. 73.5 mmol) were added.
• the resulting mixture was sti ⁇ ed at room temperature for 5 hr.
• the resulting solution was concentrated and the residue was treated with water (300 mL).
• the slurry was sti ⁇ ed for 2 hr and then filtered. The cake was washed with water, dried in an oven at 40 °C under house vacuum overnight.
• Example 43 Synthesis of [4-(5-Methylthio-3-pyridin-3-yl-nyrazol-l-vQphenyll-(2- chloro-6-fluorobenzyl)amine.
• the reaction mixture was diluted with ice-water, the aqueous layer was extracted with ethyl acetate and the combined organic extracts were washed with water and dried (MgS0 ).
• the resulting dimethylthiovinyl ketone (5.2 g) was used directly in the next step.
• L is -NHC(S)NH-
• L is -NHCH
• L is -NHCH
• L is -NHCH
• L is -NHCH
• L is -NHCH
• L is -NHCH
• L is -NHCH
• the IL-2 promoter assay measures transcriptional activation of a luciferase reporter gene which has been placed under control of the IL-2 promoter/enhancer. All the known regulatory features of the IL-2 gene are contained within a -300 bp sequence immediately upstream of the open reading frame. The region -328 to +35 relative to the transcription start site of the IL-2 gene is obtained by RT-PCR of human genomic DNA and is subcloned into the promoterless luciferase reporter vector pGL2-Basic (Promega). The resulting construct, pIL2P-luc.
• pcDNA/Neo a vector containing a neomycin resistance gene, pcDNA/Neo (Invitrogen)
• Jurkat cells a human T cell line
• pcDNA/Neo a vector containing a neomycin resistance gene
• the cells are pelleted by centrifugation, washed once with PBS, resuspended in RPMI (phenol red-free) containing 5% FBS, and dispensed into 96-well, white microtiter plates (Packard) at 50,000 cells/well.
• the cells are pre-incubated with compounds (1 ⁇ g/ml) for 15 min prior to addition of ionomycin (1 ⁇ g/ml) and PMA (10 ng/ml) in a final volume of 100 ⁇ l.
• Luc-Lite lysis buffer/luciferase assay buffer (Promega) is added and luminescence measured using a Packard TopCount scintillation counter/luminometer.
• Human peripheral blood is obtained from healthy donors by venipuncture and the mononuclear cell fraction is prepared by centrifugation on Ficoll Hypaque (Phamacia) density gradients. Contaminating red blood cells are lysed and the CD3+/CD4+ cells are purified using immunoaffinity columns (R&D Systems or CellPro). The cells are resuspended and dispensed in 96 well microtiter plates. Test compounds are added to the cells approximately 15 minutes prior to stimulation with ionomycin (1 ⁇ g/ml) and PMA (10 ng/ml). The final volume of the assay is 100 ⁇ L.
• the cells are pelleted by centrifugation, and the supernatants are collected and stored at -70°C until assayed for IL-2 using a commercial ELISA kit (Genzyme).
• the ability of cells to recognize other cells from self or from another genetically different individual (non-self) is an important property in maintaining the integrity of tissue and organ structure.
• the allogeneic cell transplant response is therefore an important model for studies of transplant rejection.
• This T-cell-mediated immune response can be induced in adult mice by the injection of lymphocytes from an histoincompatible mouse strain. This response is characterized by T cell proliferation which is limited to the popliteal lymph node that receives drainage from the footpad area. No in vitro system exists that can exactly duplicate completely this in vivo response.
• the assay is commonly used to evaluate new and novel potential immunosuppressive molecules. The assay is preferred to the local GVH response in mice because the magnitude of the response is significantly greater (Kroczek et al., Immunology, 139. 3597 (1987)).
• mice Male or female mice (20-26 grams). Any histoincompatible mouse strains suffice for donor and recipient populations. Typically DBA mice are used as donors and C57B1/6 mice are used as recipients. A minimum of 1 week stabilization and conditioning period is usually required before use of the mice. Each study utilizes approximately 36 recipient mice divided into groups of 6. Previous studies suggest this is the minimum number of animals which yields statistically significant results.
• Donor mice are sacrificed by CC asphyxiation and spleens are removed and a cell suspension is prepared.
• the cell suspension (1.0 x lO '/metatarsal in 0.05 ml) is injected I.D. into the dorsal metatarsal skin of recipient mice.
• the animals are sacrificed by CO2 asphyxiation and the popliteal nodes are removed and weighed.
• Groups of mice receiving putative immunosuppressive agents are dosed subcutaneously, intraperitoneally or orally one hour prior to cell injection and daily thereafter. The tests last approximately four days.
• the assay involves no footpad swelling and only a moderate increase in the size of the popliteal lymph node.
• the Student's t test is used to determine significant differences between popliteal lymph nodes of groups of untreated mice and those mice treated with putative immunosuppressive agents.

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