AU4478299A - Tri-substituted imidazoles having multiple therapeutic properties - Google Patents

Tri-substituted imidazoles having multiple therapeutic properties Download PDF

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AU4478299A
AU4478299A AU44782/99A AU4478299A AU4478299A AU 4478299 A AU4478299 A AU 4478299A AU 44782/99 A AU44782/99 A AU 44782/99A AU 4478299 A AU4478299 A AU 4478299A AU 4478299 A AU4478299 A AU 4478299A
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Jerry Leroy Adams
Thimothy Francis Gallagher
Ravi Shanker Garigipati
Peter William Sheldrake
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SmithKline Beecham Corp
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SmithKline Beecham Corp
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Description

AUSTRALIA
PATENTS ACT 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT
(ORIGINAL)
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ft. ft f ft Name of Applicant: Actual Inventors: Address for Service: SmithKline Beecham Corporation ADAMS, Jerry Leroy, SHELDRAKE, Peter William GALLAGHER, Timothy Francis AND GARIGIPATI, Ravi Shanker DAVIES COLLISON CAVE, Patent Attorneys, 1 Little Collins Street, Melbourne, 3000 Tri-substituted imidazoles having multiple therapeutic properties Invention Title: The following statement is a full description of this invention, including the best method of performing it known to us: Q:\OPER\PDB\2209391.239 27/8/99 P:\OPER\I'DB\2209391.239 27/8/99 TRI-SUBSTITUTED IMIDAZOLES HAVING MULTIPLE THERAPEUTIC PROPERTIES This application is a divisional of application 71850/98, the entire contents of which is incorporated herein by reference.
This invention relates to a novel group of imidazole compounds, processes for the preparation thereof, the use thereof in treating cytokine mediated diseases and pharmaceutical compositions for use in such therapy.
BACKGROUND OF THE INVENTION Interleukin-1 (IL-1) and Tumor Necrosis Factor (TNF) are biological substances produced by a variety of cells, such as monocytes or macrophages. IL-1 has been demonstrated to mediate a variety of biological activities thought to be important in immunoregulation and other physiological conditions such as inflammation [See, Dinarello et al., Rev. Infect Disease, f, 51 (1984)]. The myriad of known biological activities of IL-1 include the activation of T helper cells, induction of fever, stimulation of prostaglandin or collagenase production, neutrophil chemotaxis, induction of acute phase proteins and the suppression of plasma iron levels.
There are many disease states in which excessive or unregulated ILproduction is implicated in exacerbating and/or causing the disease. These include rheumatoid arthritis, osteoarthritis, endotoxemia and/or toxic shock syndrome, other S. acute or chronic inflammatory disease states such as the inflammatory reaction induced by endotoxin or inflammatory bowel disease; tuberculosis, aterosclerosis, muscle degeneration, cachexia, psoriatic arthritis, Reiter's syndrome, rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis, and acute synovitis. Recent evidence also links IL-1 activity to diabetes and pancreatic B cells.
Dinarllo, J. Clinical Immunology, 5 287-297 (1985), reviews the biological activities which have been attributed to IL- 1. It should be noted that some of these effects have been described by others as indirect effects of IL- 1.
Excessive or unregulated TNF production has been implicated in mediating or exacerbating a number of diseases including rheumatoid arthritis, rheumatoid spondylitis. osteoarthritis, gouty arthritis and other arthritic conditions; sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distress syndrome, cerebral malaria, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcoisosis, bone resorption diseases, reperfusion injury, graft vs. host reaction, allograft rejections, fever and myalgias due to infection, such as influenza, cachexia secondary to infection or malignancy, cachexia, secondary to acquired 2 immune deficiency syndrome (AIDS), AIDS. ARC (AIDS related complex), keloid formation, scar tissue formation, Crohn's disease, ulcerative colitis, or pyresis.
AIDS results from the infection of T lymphocytes with Human Immunodeficiency Virus (HIV). At least three types or strains of HIV have been identified, HIV-1, HIV-2 and HIV-3. As a consequence of HIV infection, T-cell mediated immunity is impaired and infected individuals manifest severe opportunistic infections and/or unusual neoplasms. HIV entry into the T lymphocyte requires T lymphocyte activation. Other viruses, such as HIV-1, HIV-2 infect T lymphocytes after T Cell activation and such virus protein expression and/or replication is mediated or maintained by such T cell activation. Once an activated T lymphocyte is infected with HIV, the T lymphocyte must continue to be maintained in an activated state to permit HIV gene expression and/or HIV replication. Monokines, specifically TNF, are implicated in activated T-cell mediated HIV protein expression and/or virus replication by playing a role in maintaining T lymphocyte activation. Therefore, interference with 15 monokine activity such as by inhibition of monokine production, notably TNF, in an HIV-infected individual aids in limiting the maintenance of T cell activation, thereby reducing the progression of HIV infectivity to previously uninfected cells which results in a slowing or elimination of the progression of immune dysfunction caused by HIV infection. Monocytes, macrophages, and related cells, such as kupffer and glial cells, have also been implicated in maintenance of the HIV infection. These cells, like T-cells, are targets for viral replication and the level of viral replication is dependent upon the activation state of the cells. [See Rosenberg A The Immunopathogenesis of HIV Infection, Advances in Immunology, Vol. 57, (1989)]. Monokines, such as TNF, have been shown to activate HIV replication in monocytes and/or macrophages [See Poli, tLal Proc. Natl. Acad. Sci., 87:782-784 therefore, inhibition of monokine production or activity aids in limiting HIV progression as stated above for T-cells.
TNF has also been implicated in various roles with other viral infections, such as the cytomegalia virus (CMV), influenza virus, and the herpes virus for similar reasons as those noted.
Interleukin-8 (IL-8) is a chemotactic factor first identified and characterized in 1987. IL-8 is produced by several cell types including mononuclear cells, fibroblasts, endothelial cells, and keratinocytes. Its production from endothelial cells is induced by IL-1, TNF, or lipopolysachharide (LPS). Human IL-8 has been shown to act on Mouse, Guinea Pig, Rat, and Rabbit Neutrophils. Many different names have been applied to IL-8, such as neutrophil attractant/activation protein-1 (NAP-1), monocyte derived neutrophil chemotactic factor (MDNCF), neutrophil activating factor (NAF), and T-cell lymphocyte chemotactic factor.
3 [L-8 stimulates a number of functions in vitro. It has been shown to have chemoattractant properties for neutrophils, T-lymphocytes, and basophils. In addition it induces histamine release from basophils from both normal and atopic individuals as well as lysozomal enzyme release and respiratory burst from neutrophils. IL-8 has also been shown to increase the surface expression of Mac-I (CDI Ib/CD18) on neutrophils without de novo protein synthesis, this may contribute to increased adhesion of the neutrophils to vascular endothelial cells. Many diseases are characterized by massive neutrophil infiltration. Conditions associated with an increased in IL-8 production (which is responsible for chemotaxis of neutrophil into the inflammatory site) would benefit by compounds which are suppressive of IL-8 production.
IL-I and TNF affect a wide variety of cells and tissues and these cytodines as well as other leukocyte derived cytokines are important and critical inflammatory mediators of a wide variety of disease states and conditions. The inhibition of these cytokines is of benefit in controlling, reducing and alleviating many of these disease states.
There remains a need for treatment, in this field, for compounds which are cytokine suppressive anti-inflammatory drugs, i.e. compounds which are capable of inhibiting cytokines, such as IL-i, IL-6, IL-8 and TNF.
SUMMARY OFTHE WMIToN •.This invention relates to the novel compounds of Formula and pharmaceutical compositions comprising a compound of Formula and a pharmaceutically acceptable diluent or carrier.
z "This invention also relates to a method of inhibiting cytokines and the treatment of a cytokine mediated disease, in a mammal in need thereof, which comprises administering to said mammal an effective amount of a compound of Formula This invention more specifically relates to a method of inhibiting the production of IL-I in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula This invention more specifically relates to a method of inhibiting the production of IL-8 in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula This invention more specifically relates to a method of inhibiting the production of TNF in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula Accordingly, the present invention provides a compound of Formula I R,
R
2
R
4 N (:N RI is 4-pyridyl, pyrimidinyl, quinolyl, isoquinolinyl, quinazolin-4-yl, 1-inmidazolyl or 1-benzimidazolyl, which heteroaryl ring is optionally substituted with one or two substituents each of which is independently selected from CI.4 alkyl. halogen, hydroxyl, Cj-4 alkoxy, CI-4 alkylthio, CI-4 alcylsulfinyl, CH2ORl1 2 NR1OR20, or an N-heterocyclyl ring which ring has from 5 to 7 members and optionally contains a.i additional heteroatom selected from oxygen, sulfur or NRI R4 is phenyl, naphth- 1 -yl or naphth-2-yl,. 'which is optionally substituted by one or two substiruents, each ofwhich is independently selected, and which, for a 4-phenyl, 4-naphth-1-yl, 5-naphth-2-yl or 6-naphth-2-yl substitiuent, is halogen, cyano, nitro, -C(Z)NR7R 17 -C(Z)0R16, -(CRIOR2O)mCOR1 2 -0R12, halo-substituted-C 1 -4 alkyl. CL-4 alkyl, -ZC(Z)R 12, -NR 1OC(Z)R16. or -(CRIOR20)mNRIOR20 and which, for other positions of ~:'.substitution, is halo, cyano, -C(Z)NR13RI 4 -C(Z)0R3, -(CRIoR20)mCOR 3 -S(O)MR3, -0R3, halo-substituted-CI- 4 alkyl, -CI-4 alkyl, -(CR1 0R20)mNRIOC(Z)R3, -NR1OS(O)MR8, -NRI OS(O)m 'NR7Rl 7, -ZC(Z)R3 or -(CRIOR20)mNR13Rp4, R2 is CI-10( alkyl N3, -(CR1OR20)n 0R9, heterocyclyl, heterocyclyiC 1-10 alkyl.
C1..ioalkyl, halo-substiutedCi.io alkyl, C2-10 alkenyl, C2-1o alkynyl, C3-7 9..20 cycloalkyl, C3-7cycloalkylC I -i1 alkyl, C5-7 cycloalkenyl, C5-7cycloalkenyl-Cl1 -alkyL, aryl, aiylCl -10 aikyL, heteroaryl, (CRIOR2O)nOR II, (CR 10R20)nS(O)mR 18.
(CR 10R20)nNHS(O)2Rl8, (CRIOR2O)nNR 13R 14, CCRIoR2O)nNO2.
(CR 10R20)nCN, (CR 10R20)n'SO2Rl8, (CR1OR2)nS(O)m'NR13Rl 4 (CRIOR2O)nC(Z)Rl1. (CRIOR2o)nOC(Z)RIl, (CRIoR2o)nC(Z)QR11, (CR 10R20)nC(Z)NR1I3R 14, (CRIoR2o)nC(Z)NRll 0R9, (CR 10R20)nNR 1OC(Z)Rl I, (CR1OR20)nNR LOC(Z)NRi 3R 14.
(CRIoR2)nN(OR6)C(Z)R1 3
RI
4 (CRIoR20)nN(0R6)C(Z)RI 1, (CR1 0R20)nC(=NOR6)RI 1, (CRi oR2O)nNR IOC(=NR 19)NR13R 14.
(CR1 0R2O)nOC(Z)NR 13R 14. (CR1 0R20)nNR 1OC(Z)NRi 3R 14.
(CP. I R20)nNRlwC(Z)OR 10, 5-(Rjg)- l.
2 4 -oxadizaol-3-yI or 4 (R){RiRi,-4,5dhydro.1,2,4oxadiazo-3.y; wherein the cycloalcyl, cycloalkyl alkyl, aryl, arylalkyl, heteroaryl, hateroaryl alkyl, heterocycic and heterocyclic alkyl groups may be optionally substituted; nfis an nteger having avalue of I o 0 n' is 0, or an integer having a value of 1 to m is 0, or the integer I or 2; Z is oxygen or sulfur; m' is I or 2, R3 is heterocyclyl, heterocyclylC I -1 alkyl or RS; is hydrogen, C 1A- alkYl, C2-4 alken3'I, C2-4 alknYIiy or NR7R 17. excluding the mocities -SR5 being -SNR7R17 and -SOR5 being -SOH; R6 is hydrogen, a pharmaceutically acceptable cation, CI-10 alkYl, C 3 7 CYCloalkyl, aryl. arylC 14 alkyl, heteroaryl, heteroarylalkyl, heterocyclyl, aroyl, or C 1 alkanoyl R7 and R 17 is each independently selected from hydrogen or C 1A- alkyl or R7 and R 17 together with the nitrogen to which they are attached form a heterocyclic ring of to 7 members which ring optionally contain an additional heteroatom selected from oxygen, sulfur or R8 is C1-l1O alkyl. halo-substituted C1-10 alkyl, C2-jo alkenyl, C2- 10alkynYl, C3-7 cycloalkyl, C5-7 cycloalkenyl, aryl, aryiC I 1 alkyl, heteroaryl, heteroarylC 1. ~alkyl, (CR 10R20)nOR 11, (CR 10R20)nS(O)mRl 8. (CR 10R20)nNHS(O)2R I8, (CRiOR2O)nNRI3RI4; wherein the aryl. arylalkyl. heteroaryl, heteroaryl. alkyl may be optionally substituted; R9 is hydrogen, -C(Z)RII or optionally substituted Ci-10 alkyl, S(O)2R18, optionally substituted aryl or optionally substituted aryl-C 1 4 alkyl; R 10 and R20 is each independently selected from hydrogen or C 14 alkyl; RIIis hydrogen, Ci..loalkyl. C3-.7 cycloalkyL, hetrocyclyl, beterocyclyl Cl-. ialkyl, aryl, aryiC 1.10alkyl,hetoaryl or heterarylCi..1()alkyl;, R12 is hydrogen or RIC Ri13 and R 14 is each independently selected from hydrogen or optionally substituted CiA- alkyl. optionally substituted aryl or optionally substituted aryl-C 1 A4 alkyl, or together with the nitrogen which they are attached form a heterocyclic ring of 5 to 7 members which ring optionally contain an additional heteroatom selected from oxygen, sulfur or NR9 is RIO or C(Z)-CIA4 alkylI; Ri 16 is Ci1- alkyl, halo-substituted-C 1-4 alkyl, or C3-7 cycloalkyl;.
R 18 is C I -10 alkyl, C3-7 cycloalkyl, heterOCYClyl arYL, arylalkyl, heterocyclyl, heterocyclyl-C I oalkyl, heteroaryl or heteroarylalkyl; R 19 is hydrogen, cyano, C 1 4 alkyl, C:3.7 cycloalkyl or aryl; or a pharmaceutically acceptable salt thereof.
DETAILED DESCRIPTON OF THE INVENTION The novel compounds of Formula may also be used in association with the veterinary treatment of mammals, other than humans, in need of inhibition of cytokine inhibition or production. In particular, cytokine mediated diseases for treatment, therapeutically or prophylactically, in animals include disease states such as those noted herein in the Methods of Treatment section, but in particular viral infections.
Examples of such viruses include, but are not limited to, lentivirus infections such as, equine infectious anaemnia virus, caprine arthritis virus, visna vinis, or maedi virus or retrovirus, infections, such as but not limited to feline immunodeficiency virus (Fly), bovine imnmunodeficiency virus, or canine immunodeficiency virus or other retroviral infections.
In Formula suitable R I moieties includes 4-pyridyl, 4-pyriruidinyl, 4quinolyl, 6-isoquinolinyl, 4-quinazolinyl, 1-inlidazolyl and 1-benziinidazolyl, of which the 4-pyridyl, 4-pyrimidinyl and 4-quinolyl are preferred. More preferred is an optionally substituted 4-pyrimidinyl and an optionally substituted 4-pyridyl, and most preferred is the an optionally substituted 4-pyriinidinyl ring.
*Suitable substituients for the R I heteroaryl rings are C 1-4 alkyl, halo, OH, C 1-4 alkoxy, Cl1-4 alkylthio. C1-4 alkylsulflnyl. CH20R12, NRlIOR2O, or an N-heterocyclyl ring which ring has from 5 to 7 members and optionally contains an additional heteroatom selected from oxygen, sulfur or NR15. A preferred substituent for allRI **moieties is C 1- alkyl, in particular methyl, and NR 1OR2O, preferably where Rio and are hydrogen or methyl, more preferably RIO and R20 are hydrogen. A more preferred, substitnent is the NR~0R2o moiety. Preferred ring placement of the Ri substituent for on the 4-pyridyl derivative is the 2-position, such as 2-methyl-4-pyridyl.
Preferred ring placement on the 4-pyrimidinyl is also at the 2-position, such as 2methyl-pyrimidine or 2-amino pyrimidine.
Suitably, R4 is phenyl, naphth- I-yi or naphth-2-yl, or a heteroaryl, which is optionally substituted by one or two substituents. More preferably R4 is a phenyl or naphthyl ring. Suitable substitutions for R4 when this is a 4-phenyl, 4-naphth- 1-yl, naphth-2-yi or 6-naphth-2-yl moiety are one or two substituents each of which are independently selected from halogen, -SR5, -S0R5, -0R6, CF3, or -(CR1OR2O)mNRiOR2O, and for other positions of substitution on these rings preferred substitution is halogen, -S(O)mR3, -0R3, CF3, -(CRiOR2O)mNRi3R 14, -NRIOiC(Z)R3 and -NRIOS(O)mR3. Preferred substituents for the 4-position in phenyl and naphth-lI-yl and on the 5-position in naphth-2-yi include halogen, especially fluoro and chloro and -SR5 and -SOR5 wherein R5 is preferably a C 1-2 alkyl. more preferably methyl; of which the fluoro and chloro is more preferred, and most especially preferred is fluoro. Preferred substituents for the 3-position in phenyl and naphth- I-yl rings include: halogen, especially fluoro and chloro; -0R3, especially C 1- 7..
aikoxy; CF3, NR1OR2O, such as amino; -NRIOC(Z)R3, especially -NIHCO(C 1 10 alkyl); -NRlOS(O)mnR8, especially -NHSO2(CI..10 alkyl), and -SR3 and -SOR3 wherein R3 is preferably a Cl-2 alkyl, more preferably methyl. When the phenyl ring is disubstituted preferably it is two independent halogen moieties, such as fluoro and chloro, preferably di-chioro and more preferably in the 3,4-position.
Preferably, the R4 moiety is an unsubstituted or substituted phenyl moiety.
More preferably, R4 is phenyl or phenyl substituted at the 4-position with fluoro and/or substituted at the 3-position with fluoro, chioro, CI1- alkoxy, inethane-sulfonarnido or acetaniido, or R4 is phenyl di-substitutect at the 3,4-position with chioro or fluoro, more Preferably chioro. Most preferably R4 is a 4-tluorophenyl.
In Formula suitably Z is oxygen.
Suitably, R2 is CI-1iO alkyl N3, -(CR10R20)n' 0R9, heterocyclyl, heterocyclylCi.io alkyl, Cl-lOalyl. halo-substituted CI10Oalkyl, C2.lO alkenyl, C2- C3-7 cycloalkyl, C3-7cYcloalkYlCl..10 alkyl, C5-7 cycloalkenyl, C5-7 cycloalkenyl Cll alkyl, axyl,arylCl-loalkyl, heteroaryl, heteroarylCl.lo alkyl, (CR1 0R20)nOR 11. (CR 10R20)nS(Q)mR I 8 (CR 10R20)nNHS(O)2RI8, (CR 10R20)nNR 13R14. (CR 10R20)nNO2, (CRI 0R20)nCN, (CR lOR20)n'SO2R 18.
(CR 10R20)nS(O)m'NR13R 14. (CR1OR2O)nC(Z)RI 1, (CR 10R20)nOC(Z)RI 1, (CR 10R20)nC(Z)OR1 1.(CR1OR2O)nC(Z)NR13R14- (CR 10R20)nC(Z)NR 11 R9, (CR 10R20)nNR iOC(Z)Ri 1.(CR1OR2O)nNR 1OC(Z)NR13Rl4, 99 (CR1OR2)nN(0R6)C(Z)NRl3RI4. (CRIOR20)nN(OR6)C(Z)R1 (CR 10R20)nC(=NOR6)R 1. (CR1OR2O)nNR iOC(=NR 19)NR1 3R 14.
(CRIOR2O)nOC(Z)NR 13RI4. (CR1OR2O)nNR1OC(Z)NR13RI4, (CR 10R20)nNR IOC(Z)ORIO, 5-(Rjg)- 1,2,4-oxadizaol-3-yl or 4-(R12)-5-(RIgRt9)-4,5-dihydro-1,2,4-oxadiazol-3-yl;wherein the aryL, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclic, and heterocyclicaklyl moieties may be optionally substituted; wherein n is an integer having a value of 1 to 10, mn is 0, or the integer 1 or 2; n' is 0, or an integer having a value of 1 to 10, and mn' is 1 or 2.
Preferably n is 1 to 4.
Preferably R2 is an optionally substituted heterocyclyl ring, and optionally substituted heterocyclyiC 1- 10 alkyl, an optionally substituted C 1- 10 alkyl, an optionally substituted C3-7cYcloalkyl, an optionally substituted C3.7cycloalkyl C 1 alkyl, (CR l0R2O)nC(Z)OR 11 group, (CR 10R20)nNR 1 3R14, (CRIOR2O)nNHS(O)2R18. (CRIOR2O)nS(O)mRl8, an optionally sub~stituted aryl; an optionally substituted aryiC 1 -10 alkyl, (CRIOR20)nOR1 I, (CR loR2o)nC(Z)R1 I, or (CR 10R20)nC(=NOR6)R I I group.
More preferably R2 is an optionally substituted heterocyclyl ring, and optionally substituted heterocyclylCi.l0 alkyl, optionally substituted aryl, (CRlIOR2O)nNR 13RI14, or (CRI oR2o)nC(Z)OR I I group.
When R2 is an optionally substituted heterocyclyl the ring is preferably a ruorphoino, pyrrolidinyl, or a piperidinyl group. When the ring is optionally substituted the substituents may be directly attached to the free nitrogen, such as in the pipenidinyl group or pyrrole ring, or on the ring itself Preferably the ring is a piperidine or pyrrole, more preferably piperidine. The heterocyclyl ring may be optionally substituted one to four times independently by halogen; CI1-4 alkyl; aryl, such as phenyl; aryl alkyl, such as benzyl wherein the aryl or aryl alkyl moieties themselves may be optionally substitued (as in the definition section below); C(O)ORi1 such as the C(O)CI1- alkyl or C(O)OH moieities; C(O)H; C(O)C 1 alkyl, *hydroxy substituted C 1- alkyl, CI1A- alkoxy, S(O)mC 1 alkyl (wherein me is 0, 1, or NRIOR20 (wherein RIO and R20 are independently hydrogen or CI-4alkyl).
Preferably if the ring is a piperidine, the ring is attached to the imidazole at the 4-position, and the substituents are directly on the available nitrogen, ie. a I-Formyl-4piperidine, 1-benzyl-4-piperidine, 1-methyl-4-piperidine, l-ethoxycarbonyl-4piperidine. If the ring is substituted by an alkyl group and the ring is attached in the 4position, it is preferably substituted in the 2 or 6 position or both, such as 2,2,6,6teamethyl-4-piperidine. Siiniarly, if the ring is a pyrrole, the ring is attached to the imidazole at the 3-position, and the substituents are aldo directly on the available "eve*'nitrogen.
When R2 is an optionally substituted beterocyclyl Ci-io alkyl group, the ring is preferably a morpholino, pyrrolidinyl, or a piperidinyl group. Preferably this alkyl S. .25 moiety is from I to 4. more preferably 3 or 4, and most preferably 3, such as in a propyl group. Preferred heterocyclic aikyl groups include but are not limited to, morpholino ethyl. morpholino propyl, pyrrollidinyl propyl, and piperidinyl propyl moieties. The heterocyclic ring herein is also optionally substiuted in a similar manner to that indicated above for the direct attachement of the heterocycyl.
When R2 is an optionally substituted C3-7cycloalkyl, or an optionally substituted C3-7cycloalkyl Cl1.j1 alkyl, the cycloalkl group is preferably a C5 to C6 ring which ring may be optionally substituted I or more times independtly by halogen, such as fluorine, chlorine, bromine or iodine; hydroxy; Cl1-jo alkoxy, such as methoxy or ethoxy; S(O)m alkyl, wherein m is 0, 1, or 2, such as methyl thio, methylsulimyl or methyl sulfonyl; amino, mono di-substituted amino, such as in the NR,7R17 group; or where the R7R 17 may cyclize together with the nitrogen to which they are attached to form a 5 to 7 membered ring which optionally includes an additional heteroatom selected from OINIS; C I- 10 alkyl, such as methyl. ethyl, propyl, isopropyl, or t-butyl; halosubstitued alkyl, such as CF3; hydroxy substituted C I oalkyl; C(O)OR 11, such as the free acid or methyl ester derivative; an optionally substituted aryl, such as phenyl; an optionally substituted arylalkyl, such as benzyl of phenethyl; and further where these aryl moieties may also be substituted one to two times by halogen, hydroxy; C I alkoxy; S(O)m alkyl; amino, mono di-substituted amino, such as in the NR7R 17 group; alkyl or halosubstitued alkyl.
When R2 is (CRIOR20)flNRl3RI4, R13 and R14 are as defined in Formula that is R13 and R14 are each independently selected from hydrogen, optionally substituted C1.4 alkyl, optionally substituted aryl or an optionally substituted aryl-C alkyl, or together with the nitrogen which they are attached form a heterocyclic ring of 5 to 7 members which ring optionally contains an additional heteroazomn selected from oxygen, sulfur or NR9. It is recognized that in some instances this can yield the same ~.:moiety as a heterocyclic C I1-10 alkyl moiety noted above which is also a suitable R2 variable. Preferably R13 and RJ4 are independently hydrogen, CI..4 alkyl, preferably methyl, or benzyL. The n term is preferably I to 4, more preferably 3 or 4, and most preferably 3, such as in a propyl group. Preferred groups include, but are not limited to, aminopropyl, (N-methyl-N-benzyl)aminopropyl, (N-Phenylmethyl)aniino. I -propyl, or diethylammno propyl.
When R2 is a (CRlOR2o)nC(Z)ORll group, RI lis suitably hydrogen, Cl-4 alkyl, especially methyl. The n term is preferably 1 to 4, mote preferably 2 or 3, such as in an ethyl or propyl group. Preferred groups include, but are not limited to, .*carboxymethyl- I1-butyl. carboxy- I1-propyl, or 2-acetoxyethyL When R2 is a (CR lOR2OMnS(O)mRlS group m isO,. 1, or 2, and RI 8 is preferably aryl, especially phenyl, or C 1- 10 alkyl. especially methyl. T1he n term is preferably 1 to 4, more preferably 2 or 3, such as in in ethyl or propyl group.
~When R2 is a(CR IOR2OMnORll group, RI is suitably hydrogen, aryl, especially phenyl, or C 1-10 alkyl, especially methyl or ethyL The n term is preferably I to 4, more preferably 2 or 3. such as in an ethyl'or propyl group.
When R2 is a (CRIOR2o)nNHS(O)2R18 group, Rlg is suitably alkyl, especially methyl. The n term is preferably 1 to 4. more preferably 2 or 3, such as in an ethyl or propyl group.
When R2 is a optionally substituted aryl. the aryl is preferably phenyl. The aryl ring may be optionally substituted one or more tunes, preferably by one or two substituents, independently selected from C 1 alkyl, halogen, especially fluoro or chioro, (CRlOR2O)tORl 1, -(CRlOR20)tNRlIOR2O, especially amino or mono- or dialkylwmino -(CRlOR2o)tS(O)reRl8, wherein m is 0, 1 or 2; SH-, -(CRloR20)DNR13Rl4, -NRIOC(Z)R3 (such -NHCO(Cj..loalkyl)); -NRIOS(O)mR8 (such as -NHSO2(C I -l1 alkyl)); t is 0, or an integer of I to 4. Preferably the phenyl is substituted in the 3 or 4- position by -(CR I 0R20)tS(O)mR I and Rl18 is Preferably CI-1j 0 alkyl, especially methyl.
When R2 is an optionally substituted heteroaryl or heteroarylalcyl group the ring may be optionally substituted one or more times, preferably by one or two substituents, independently selected from one or more times, by Cl1 -4 alkyl, halogen, especially fluoro or chioro, (CRI0R2O)@OR1 1, -(CRIoR2O)tNRIOR20, especially amino or mono- or di-alkylamio -(CR 10R20)tS(O)mRl8, wherein mn is 0, 1 or 2 SH-, -(CRIoR2o)n-NRI3RI4, -NRIOC(Z)R3 (such -NHCO(CI..iO alkyl)), -NRIOS(O)mR8 (such as -NHSO2(Cl-I0 alkyl)); t is 0, oran integer of 1 to 4.
One skille in the art would readily recognize that when R2is a (CR loR2o)nOC(Z)Rl 1, or (CRlOR2)nOC(Z)NR13RI4 moiety, or any similarly substituted group that n is preferably at least 2 which will allow for the synthesis of stable compounds.
Preferably R2 is a C 1 -4 alkyl (branched and unbranched), especially methyl, a methylthio propyl, a methylsulfinyl propyl. an amino propyl, N-methyl-N-benzylamzino propyl group, diethylamino propyl, cyclopropyl methyl, morpholinyl butyl, morpholinyl propyl, a morpholinyl ethyl, a piperidine or a substituted piperidine.
More preferably R2 is a methyl, isopropyl, butyl, t-butyl, n-propyl, methyithiopropyl, -:or methyLsullinyl propyl, morpholino propyl, morpholinyl butyl, phenyl substituted by halogen, tiaikyl or sulfinyl alkyl such as a methylthia, methylsulfinyl or methylsulfonyl moiety; piperidinyl, 1 -Formyl-4-pipenidine, Il-benzyl-4-piperidine, I methyl-4-piperidine, or a I -ethoxycarbonyl-4-piperidine.
In all instances herein where theme is an alkenyl or alkynyl moiety as a substituient group, the unsaturated linkage, iLe., the vinylene or acetylene linkage is preferably not directly attached to the nitrogen, oxygen or sulfur moieties, for instance in 0R3, or for certain R2 moieties.
As used herein, "optionally substituted" unless specifically defined shall mean such groups as halogen, such as fluorine, chlorine, bromine or iodine; hydroxy; hydroxy substituted C 1. oalkyl; C 1-l1 alkoxy, such as methoxy or ethoxy, S(Q)m alkyl, wherein mn is 0, 1 or 2. such as methyl thio, methylsulfinyl or methyl sulfonyl; amino, mono di-substituted amino, such as in the NR7R17 group; or where the R7RI7 may together with the nitrogen to which they are attached cyclize to form a 5 to 7 membered ring which optionally includes an additional heteroatom selected from OINIS; Cl-l10alkyl, cycloalkyl, or cycloalkyl alkyl group, such as methyl, ethyl, propyl, isopropyl, t-butyl. etc. or cyclopropyl methyl; halosubstituted C 1- 10 alkyl, such CF3; an optionally substituted aryl, such as phenyl, or an optionally substituted ary~kalkyl, such as benzyl or phenethyl, wherein these aryl moieties may also be substituted one to two times by halogen; hydroxy; hydroxy substituted alkcyl; C 1-10o alkoxy; S(O)m alkyl;, amino, mono di-substituted amino, such as in the NR7R17 group; alkyl, or CF3.
1 1.- In a preferred subgenus of compounds of Formula RI is 4-pyridyl, 2-ailcyl- 4-pyridyl, 4-quinolyl, 4-pyrinidinyl, or 2-amino-4-pyrimidinyl, R2 is mOrpholinyl propyl, aminopropyl, piperidinyl, N-benzyl-4piperidine, or a N-methyl-4-piperidine; S and R4 is phenyl or phenyl substituted one or two times by tluoro, chioro, C 1 alkoxy, -S(O)m alkyl, rnezhanesulfonamido or acetamido.
A preferred subgrouping of compounds of Formula are those where R2 is other than methyl when RI1 is pyridyl, and R4 is an optionally substituted phenyl.
Suitable pharmaceutically acceptable salts are well known to those skilled in the art and include basic salts of inorganic and organic acids, such as hydroc:ilonic acid, hydrobromic acid, sulphuric acid, phosphoric acid, methane suiphonic acid, ethane sulphonic acid, acetic acid, maiic acid, tartaric acid, citric acid, lactic acid, oxalic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid, phenylacetic acid and mandelic acid. EI addition, pharmaceutically. acceptable salts of compounds of.
Formula may also be formed with a pharmaceutically acceptable cation, for instance, if a substituent group comprises a carboxy moiety. Suitable pharmaceutically ~acceptable cations are well known to those skilled in the art and include alkaline, alkaline earth, ammonium and quaternary amimonium cations.
The following terms, as used herein, refer to: -"halo" or "halogens", include the halogens: chioro, fluoro, bromno and iodo.
"C I lalkyl" or *alkyl* both straight and branched chain radicals of I to carbon atoms, unless the chain length is otherwise limited, including, but not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl and the like.
The term "cycloalkyl" is used herein to mean cyclic raicals, preferably of 3 to 8 carbons, including but not limited to cyclopropyl, cyclopentyl, cyclohexyl, and the like.
The term "cycloalkenyr" is used herein to mean cyclic radicals, preferably of to 8 carbons, which have at least one bond including but not limited to cyclopentenyl, cyclohexenyl, and the lie.
The term "alkenyl" is used herein at all occurrences to mean straight or branched chain radical of 2-10 carbon atoms, unless the chain length is limited thereto, including, but not limited to ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1butenyl. 2-butenyl and the like.
"aryl" phenyl and naphthyl; 12 "'eeo heteroaryl' (on its own or in any combination, such as "heteroaryloxy-, or "htraryl alkyl") a 5-10 membered aromatic ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, 0 or S, such as, but not limited, to pyrrole, pyrazole, furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine, pyrimidine, oxazole, thiazole, thiadiazole, triazole, inudazole, or benzimidazoje.
heteroyclic" (on its own or in any combination, such as nheterocyclylalkyl") -a saturated Or Partially unsaturated 4-10 memnbered ring system in which one or more rings contain one or more heteroatoms selected from the group consisting of N, 0, or S; such as, but not limited to, pyrrolidine, piperidine, piperazine, morpholine, tetrahYdro'pyran, or imidazolidine.
he term "aralkyl" or "heteroarylalkyl" or wheterocycicalkyl" is used herein 60to mean C 1-4 alkyl as defined above attached to an aryl, heteroazyl or heterocyclic moiety as also defined herein unless otherwise indicate.
*"sulflnyl* the oxide S of the correponding sulfide, the tem thiorefers to the sulfide, and the term "sulfonyl" refers to the fully oxidized S (0)2 moiety.
"aroyl" a C(O)Ar, wherein Ar is as phenyl, naphthyl, or aryl alk derivative such as defined above, such group include but are note limited to benzyl and phenethyl.
*"alkanoyl" a C(0)CI..iO alkyl wherein the alkyl is as defined above.
The compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. All of these compounds are included within the scope of the present invention.
Exemplified compounds of Formula include: 1-3(-opoiy~rpl--4fuoohnl--4prdliiaoe 1-( 3 -Chloropropyl)-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; l-(3-Azidopropyl)-4-(4-fluorophenyl)-5-(4-pyridyl)imiiazolc;.
1-(3-Aminopropyl)-4-(4-fluorophenyl)-5-(4-pyridyl)irnidazole; 1 -(3-Methylsulfonamidopropyl)-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; l-[3-(N-Phenylmethyl)aminopropyll-4-(4-fluorophenyl)-5-(4.pyridyl)imidazole; I -[3-(N-Phenylmethyl-N-methyl)aminoprpyl)4(4-fluorophenyl)..s(4pyridyl)imidazole; 1 1 (-izyaiorpl--4furohnl--4prdliiaoe l-Piperidinyl)propyl]-4-(4-fluoropheny)--(4-pyridy)imzidazole; l-[3-(Methylthio)propyl-(4-fluorophenyl)-5-(4-pyridyl)imidazoie; 1-2(-opoiy~tyl4(-loohnl--4prdliiaoe 13.
1- [3-(4-MorpholinyI)propyll-4-(3-metlylthiophenyl)-5-(4-pyridyl)inidazole; I 3(-opoiy~poyl4(-ehlsliypey)5(4prdliiaoe 1 -[3-(N-rnethyl-N-benzyl)wminopropyll-4-(3-methylthiophenyl)-5-(4pyridyl)imidazole; 1 -[3-(N-methyl-N-benzyl)aminopropylj-4-(3-methylsulfmylphenyl)--4.
pyridyl)imidazole; 1-[-(Methylthio)phenylJ-4-(4-fluorophriyl)-5-(4-pyridyl)imidazole; I -[4-{Methylsulfinyl)phenyl]-4-(4-fluorophenyl)-5-(4-pyridyl)iniidazole; I -[3-(Medyltiio)phnyl]-4-(4-fluorphenyl)-5-(4-pyridyl)imidazole; 1 -[3-(Methylsulfinyl)phenylJ-4-(4-fluorophenyl)-5-(4-pyridyl)imidazoe, 1 -[2-(Methylthio)phenyl]-4(4-fluorophenyl)-5-(4-pyridyl)inidazole; 1 -[2-(Mezhylsullinyl)phenylJ-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; 1 (4-Morpholinyl)butylJ-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; I -Cyckopropyl-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; 1 -Isopropyl-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; 1 -Cyclopropylmethyl-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; 1 -tert-Butyl-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; 1-(2,2-Diethoxyethyl)-4-4-fluorophenyl)-5-(4-pynidyl)imidazole; 1-Fonmyknethyl-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; 1 -Hydroxyiminylmethyl-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; 1 -Cyanomehyl-4-(4-fluowphenyl)-5-(4-pyridyl)imidazole; S1 -[3-(4-Morpbolinylpropyl)4(4-fluorophenyl)--(2-methylpyrid-4-yl)imidazole; 4-(4-Fluorophenyl)- 1-[3-(4-morpholinyl)propyll-5-(2-chloropyridin-4-yl) imidazole; 4-(4-Fluorophenyl)- 1-[3-(4-morpholinyl)propylJ-5-(2-amino-4-pyridinyl) imidazole; -(4-Carboxymethyl)propyl-4-(4-fluorophenyl)-5-(4-pyridyl)imidazol; 1 -(4-Carboxypropyl)-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole; 1 -(3-Carboxymediyl)ethyl-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole, 1 -(3-Carboxy)ethyl-4-(4-fluorophenyt)-5-(4-pyridyl)imidazole; 1-Benzylpiperidin-4-yI)-4-(4-fluoropbenyl)-5(4-pyridyl) imiclazole; 5-(2-Aminopyrimidin-4-yl)-4-4-fluorophenyl)- 1-(3-(4-Morpholinyl)propyl] iznidazole; 5-(2-Aminopyrunidin-4-yl)-4-(4-fluorophenyl)- I-benzylpiperidin-4-yl)imidazole; 5-(2-Aminopyrimidin-4-yI)-4-(4-fluorophenyl)- 1-(2-propyl)imidazole;, 5-(2-Aminopyrimidin-4-yl)-4-(4-fluorophenyl)- I-(cyclopropylmethyl)imnidazole; 5-(2-Aminopyrimidin4-y)--4-(4-fluoropheriyl)- 1-carboxyethyl-4piperidinyl)imidazole; 5-(2-Aminopyrinidin-4-yl,)-4-(4-fluorophenyl)- 1-(4-piperidinyl) imidazole; 1 -Methyl-4-phenyi'-5-(4-pyridyl)iznidazole; 1-Methyl-4-[3-(chlorophenyl)]-5-[4-pyridinyllimidazole; I -MethyI-4-(3-methylthiophenyl)-5-(4-pyridyl)imidazole; 14.
1 -MethyI-4-(3-methylsufnypheny)-5-(4-pyidyl)midazole; (+/-)-4-(4-Fluorophenyl)- l-( 3 -(mechylsulfmnyl)propyj-5-(4pyrdiny)midazole; 4-(4-Fluorophenyl)- I -[3mtysloy~rpl--4prdnliiaoe
I-[
3 4 -Morhoiny)propyII(4fluoropheny)(qinoly)iidaz1e; -3Peys~ypoy)--4furpey)5(4Prdniiiaoe 4-[4-(4-Rluorophenyl)- l-[ 3 4 -morphoinyl)propy1I-5-(pyrimid..2.one4-y)imiidazole; iniidazole; 4-4-Fluorophenyl)-5-[2-(methylsulfmyl).4.pynimidinyl.. -[3-(4-morpholinyl)propyljimidazole; k(IkPropeny1)-4-(4-fluorophenyl)-5-(4-pyridinyl)imidazole; I -(2-Propenyl)1-(4-fuoropeny)--(4-pyridiny)imidazD* 5-[(2-NNDimehyamino)pyimidn4yIJ-4-(4fluoropheny)- 1-[3-(4-morpholinyl)propyljimidazole; 3 -(4-Morphoflyl)propy1-5-(4-pyidinyl)-4-[3(rruoromuhy)pheyljmcjazole 1 .(Cyclopropymhy14-(3,4dthoropheny1.5(pyidinyimidawl; 1 -(CyclopropyImethy)-4-(4-fluorophenyl)-5(2me Jvypyrid..iyl)jj~dazole; 1 -[3-(4-Morphony)propy]-5-(4-pyrdiny)4(3,S-bis uoromethylpley)imidazole 5-[4-(2-Aminopyrimidinyl)J-4-(4-fluorophenyl)- 1 -(2-carboxy-2,2dimethylethyl)imidazole; 1 -onl-ieiiy)--4furpey)5(4prdnliiaoe 5-(2-Amino-4-pyriznidinyl)-4-(4-fluorophenyl). 1-(1 -rethyl-4-piperidinyl)imidazole; 1 (,-iehl3mrhli--lpoy--4fuoohnl--2Aio4 pyrunidinyl)imidazoie; 4-(-uorophny)-5-(4-pyridyl)- l-(2-acetoxyethyl)imidazole.
5-(2-Aminopyrimidin-4-yl)-4-(4-tluorophenyl)- 1-(1 -benzylpyrrolin-3-yl)irnidazole; 5-(2-Aminopyrimidin-4-y1)-4-(4-fluorophenyl)- 1 -(2.2,6,6-WAsmethylpipexidin-4- YI)iznicazole.
Preferred compounds of Formula include: 5-f4-(2-Amino)pyrimidinyl]j-4-(4-fluorophenyl)- 1-(4-N-m rpholino- 1propyl)imidazole; 4 -(2-Aminopyrimidinyl)-4-(4-fluorophenyl)- -benzyl-4-piperidinyl)imidazole; 5-( 2 -Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1 4 -piperidinyl)imidazole; 2 -Amino-4-pyrimidiny)-4-(4-fluorophenyl)-1-(1 -methyl-4-piperidinyl)imidazole.
Another aspect of the present invention is the compound, 4-phenyl-5-(4pyridyl)imidazole. Another aspect is of the present invention is a pharmaceutical composition comprising a carrier or diluent and effective amount of 4-pheny!-5-f4pyridyl)imidazole. Yet another aspect of the present invention is the novel method of treating a cytokine mediated disease state, in a mammal in need thereof, with an effective amount of 4-phenyl-5-[4-pyridyl)imidazole.
15 For purposes herein the dosage ranges, formulation details, and methods of making are analogous to the compounds of Formula The compounds of Formula may be obtained by applying synthetic procedures, some of which are illustrated in Schemes I and V. The synthesis provided for in these Schemes is applicable for the producing compounds of Formula having a variety of different Ri, R 2 and R4 groups which are reacted, employing optional substituents which are suitably protected, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, in those cases, then affords compounds of the nature generally disclosed. Once the imidazole nucleus has been established, further compounds of Formula may be prepared by applying standard techniques for functional group interconversion, well known in the art.
For instance: -C(O)NR 13Rl4 from -CO2CH3 by heating with or without catalytic metal cyanide, e.g. NaCN, and HNRI3RI4 in CH30H; -OC(O)R3 from -OH with CIC(O)R3 in pyridine; -NRlO-C(S)NRI3RI4 from -NHRIO with an alkylisothiocyante or thiocyanic acid; NR6C(O)OR6 from -NHR6 with the alkyl chloroformate; -NRIOC(O)NR13R1 4 from -NHR10 by treatment with an isocyanate, e.g. HN=C=O or RION=C=O; -NR 0-IOC(O)Rg from -NHR 10 by treatment with Cl- C(O)R3 in pyridine; -C(=NRIO)NRl3RI4 from -C(NR13RI4)SR3 with H3NR3+OAcby heating in alcohol; -C(NR13RI4)SR 3 from -C(S)NRl3RI4 with R6-I in an inert solvent, e.g. acetone; -C(S)NR13R 14 (where Ri3 or R14 is not hydrogen) from -C(S)NH2 with HNRI3Rl4-C(=NCN)-NRl 3
RI
4 from -C(=NR 13RI4)-SR3 with NH2CN by heating in anhydrous alcohol, alternatively from -C(=NH)-NRI3RI4 by treatment with BrCN and NaOEt in EtOH; -NR10-C(=NCN)SR8 from -NHRIO by treatment with (R8S)2C=NCN; -NRIOSO2R3 from -NHRIO by treatment with 16.
CISO2R3 by heaing in pyridine; -NRIOC(S)R3 from -NRIOC(O)R 8 by treatment with Lawesson's reagent [2,4-bis(4-metboxyphenyl)- I 3 2 4 -ditduadiphosphetane-2,4disulfidel; -NRIOS02CF3 from -NHR6 With triflic anhydride and base wherein R3, R6, RI, R13 and R14 are as defined in Formula herein.
R
4 CHO ArS(OH I
H
2
NCHO
CHC13 NaOH
CH
2
CI
2
H
2 0 F'rC
R
4
CH
2
NH
2
(VIII)
F\ yladn agent
R
4
CH
2
NHCHO
dehydraing agent
R
4
CH
2 NC (VI) 0@ es o 0@ 0*
S
e S.
S. S S Ar-S(O)
(IV)
NHCHO
Ar S(0) 2
L,
wherein p4 I dehydrating agent RICHO RNH2 Ar-S(O R4 NC R1N"NR 2
(M)
SCEME
Referring to Scheme I the compounds of Formula are sui'ably prepared by reacting a compound of the Formula with a compound of the Formula (III) wherein p isO, 1 or 2. R1, R2 and R4 are as definedherein,orare precrsorsof the groups R 1
R
2 and R 4 and Ar is an optionally substituted phenyl group, and thereafter if necessary converting a precursor of R 1
R
2 and R 4 to a group R 1
R
2 and R 4 17.
Suitably Ar is phenyl optionally substituted by Ci I.4alkyl, C 1 4 alkoxy or halo.
Preferably Ar is phenyl or 4-ruethyiphenyl. Suitably the reaction is performed at ambient temperature or with cooling -50* to 100 in an inert solvent such as methylene chloride, tetrahydrofuran, toluene or dimethoxyethane in the presence of an appropriate base such as 1,8-diazabicyclo undeC-7-ene (DBU) or a guanidine base such as 1,5,7- triazabicyclo (4.4.01 dec-5-ene (TBD). Suitably p is 0 or 2, preferably p is 0 in which cas the intermediates of formula (11) are very stable and capable of storage for a long time.
Precursors of the groups R 1, R2and R4 can be other R 1
R
2 and R4 groups which can be interconverted by applying standard techniques for functional group interconversion. For example a compound of the formula wherein R2 s haO -substituted C 1 1 0 alkyl can be converted to the corresponding C 1 10 alkyIN 3 derivative by reacting with a suitable azide salt, and thereafter if desired can be reduced to the corresponding C I -IOaikylNH2 compound, which in turn can be reacted with R 18 S(0) 2 X wherein X is halo chioro) to yield the corresponding
C
1 10 alkylNHS(O) 2
R
1 8 compound.
Alternatively a compound of the formnula where R 2 is halo-substituted 22 can be reacted with an amine R 13
R
1 4NH to yield the corresponding C l..l-alkYlNRl3RI4 compound, or can be reacted with an alkali metal salt of R18SH to yield the corresponding Cl..loalkylSR18 compound.
*0**In a further aspect the present invention provides compounds of the Formula @000 (1I) as defined herein with the proviso that when ArS(O)p is tosyl, then R4 is not an 'Mubtiuedoponds of Formula in Scheme 1, may be prepared by the methods of Van Leusen et al., JOC, 42, 1153 (1977). For example a compound of the Formula (D1) may be prepared by dehydrating a compound of the Formula (IV) wherein Ar, R4 arid p are as defined herein.
Suitable dehydrating agents include phosphorus oxychloride. oxalyl chloride or tosyl chloride in the presence of a suitable base such as triethylamine.
The compounds of formula (IV) may be prepared by reacting a compound of the formula R 4 CHO where R 4 is as defined herein, with ArS(0)pH and formamide under dehydrating conditions at ambient or elevated temperatur e.g. 300 to 150W, conveniently at influx, optionally in the presence of an acid catalyst. Alternatively trimethysilyichloride can be used in place of the acid catalyst. Examples of acid catalysts include camphor-lO-sulphonic acid, p-toluenesulphonic acid, hydrogen chloride or sulphuric acid.
Tie comupounds of the formula where p is 2 may also be prepared by reacting in dhe presence of a strong base a compound of the formula (VI) R 4
CH
2
NC
18..
with a compound of the formula (VII) ArSO2L1 wherein R 4 and Ar are as defined herein and LI is a leaving group such as halo, e.g. fluoro. Suitable strong bases include alkyl lithiums such as butyl lithium or lithium diisopropylamide.
The compounds of formula (VI) may be prepared by reacting a compound of the formula (VII) R 4
CH
2
NH
2 with an alkyl formate ethylformate) to yield an intermediate amide which can be converted to the desired isonitrile by reacting with a dehydrating agent such as oxalylchloride, phosphorus oxychloride or tosyl chloride in the presence of a suitable base such as triethylamine.
Alternatively a compound of the formula (VIII) may be converted to a compound of the formula (VI) by reaction with chloroform and sodium hydroxide in aqueous dichloromethane under phase transfer catalysis.
The compounds of the formula (II) may be prepared by reacting a compound of the formula RICHO with a primary amine R 2
NH
2 The amino compounds of the formula (VIII) are known or can be prepared from 15 the corresponding alcohols, oximes or amides using standard functional group interconversions.
In a further process, compounds of Formula may be prepared by coupling a suitable derivative of a compound of Formula (IX):
R
T,
N
4 X) wherein TI is hydrogen and T4 is R4, or alternatively TI is RI and T4 is H in which RI, R2 and R4 are as hereinbefore defined; with: when TI is hydrogen, a suitable derivative of the heteroaryl ring R H, under ring coupling conditions, to effect coupling of the heteroaryl ring RI to the imidazole nucleus at position 5; (ii) when T4 is hydrogen, a suitable derivative of the aryl ring R4H, under ring coupling conditions, to effect coupling of the aryl ring R4 to the imidazole nucleus at position 4.
Such aryl/heteroaryl coupling reactions are well known to those skilled in the art. In general, an organometallic synthetic equivalent of an anion of one component is coupled with a reactive derivative of the second component, in the presence of a suitable catalyst The anion equivalent may be formed from either the imidazole of Fotnula in which case the aryl/heteroaryl compound provides the reactive derivative, or the aryl/heteroaryl compound in which case the imidazole provides the reactive derivative. Accordingly, suitable derivatives of the compound of Formula (IX) 19..
or the aryl/heteroaryl rings include organornetallic derivatives such as organomagnesiuni, organozinc, organoscannane and boronic acid derivatives and suitable reactive derivatives include the bromo, iodo, fluorosulfonate and wrifluoromethanesulphonate derivatives. Suitable procedures are described in WO 91/19497, the disclosure of which is incorporated by reference herein.
Suitable organomagnesium and organozific derivatives of a compound of Formula (IX) may be reacted with a halogen, fluorosulfonate or afflate derivative of the heteroaryl or aryl ring, in the presence of a ring coupling catalyst, such as a palladium or palladium (II) catalyst, following the procedure of Kumada et al., Tetrahedron Letters, 22, 5319 (198 Suitable. such catalysts include tetraids- (triphenylphosphine)palladium and PdOl2(l,4-bisv-(diphenylphosphino)-bututeJ, .:optionally in the presence of lithium chloride and a base, such as triethylamnine. In addition, a nickel (II) catalyst, such as Ni(II)C2(1,2-biphenylphosphino)ethane, may also be used for coupling an aryl ring, following the procedure of Pridgen, J Org Chem, 1982, 47, 4319. Suitable reaction solvents include hexamethylphosphor-amide. When the heteroaryl ring is 4-pyridyl, suitable derivatives include 4-bromo- and 4-jodopyridine and the fluorosulfonate and trifte esters of 4-hydroxy pyridine. Similarly, suitable derivatives for when the aryl ring is phenyl include the brotno, fluorosulfonate, triflate and, preferably, the iodo-derivatives. Suitable organomagnesiumn and organozinc derivatives may be obtained by treating a compound of Formula (IX) or the bromo derivative thereof with an alkyllithuar compound to yield the corresponding lithium reagent by deprotonation or transmetallation, respectively. This lihium intermediate may then be trated with an excess of a magnesium halide or zinc halide to yield the corresponding organometallic reagent.
A trialkyltin derivative of the compound of Formula may be treated with a bromide, fluorosulfonate, triflate, or. preferably, iodide derivative of an aryl or heteroaryl ring compound, in an inert solvent such as tetrahydrofuran preferably containing 10% hexamethylphosphoramide. in the presence of a suitable coupling catalyst, such as a palladium catalyst, for instance tetrakis- (triphenylphosphine)palladium. by the method described in by Stike J Amer Chem Soc. 1987, 109, 5478, US Patents 4,719,218 and 5,002,942, or by using a palladium (HI) catalyst in the presence of lithium chloride optionally with an added base such as triethylamine, in an inert solvent such as dimethyl formamide. Thialkyltin derivatives may be conveniently obtained by metallaton of the corresponding compound of Formula (IX) with a lithiating agent, such as s-butyl-lithium or n-butyllithium, in an ethereal solvent, such as tetrahydrofuran, or treatment of the bromo derivative of the corresponding compound of Formula (EX) with an alkyl lithium, followed, in each case, by treatment with a trialkyltin halide. Alternatively, the bromo- derivative of a compound of Formula (IX) may be treated with a suitable heteroaryl or aryl trialkyl tin compound in the presence of a catalyst such as tetrakis-(triphenyl-phosphine)palladium, under conditions similar to those described above.
Boronic acid derivatives are also useful. Hence, a suitable derivative of a compound of Formula such as the bromo, iodo, triflate or fluorosulphonate derivative, may be reacted with a heteroaryl- or aryl-boronic acid, in the presence of a palladium catalyst such as tetrakis-(triphenylphosphine)-palladium or PdCI2l,4-bis- (diphenyl-phosphino)-butane] in the presence of a base such as sodium bicarbonate, under reflux conditions, in a solvent such as dimethoxyethane (see Fischer and Haviniga, Rec. Trav. Chim. Pays Bas, 84,439,1965, Snieckus, Tetrahedron Lett., 29,2135, 1988 and Terashimia, Chem. Pharm. Bull., 11, 4755, 1985). Nonaqueous conditions, for instance, a solvent such as DMF, at a temperature of about 100 0 C, in the presence of a Pd(I) catalyst may also be employed (see Thompson W J et al, J Org Chem, 49, 5237, 1984). Suitable boronic acid derivatives may be prepared by treating the magnesium or lithium derivative with a trialkylborate ester, such as 15 triethyl, tri-iso-propyl or tributylborate, according to standard procedures.
In such coupling reactions, it will be readily appreciated that due regard must be exercised with respect to functional groups present in the compounds of Formula (IX).
Thus, in general, amino and sulfur substituents should be non-oxidised or protected.
Compounds of Formula (IX) are imidazoles and may be obtained by any of the 20 procedures herein before described for preparing compounds of Formula In particular, an a-halo-ketone or other suitably activated ketones R4COCH2Hal (for compounds of Formula (IX) in which TI is hydrogen) or R1COCH2Hal (for compounds of Formula (IX) in which T4 is hydrogen) may be reacted with an amidine of the formula R2NH-C=NH, wherein R2 is as defineA in Formula or a salt thereof, in an inert solvent such as a halogenated hydrocarbon solvent, for instance chloroform, at a moderately elevated temperature, and, if necessary, in the presence of a suitable condensation agent such as a base. The preparation of suitable a-halo-ketones is described in WO 91/19497. Suitable reactive esters include esters of strong organic acids such as a lower alkane sulphonic or aryl sulphonic acid, for instance, methane or p-toluene sulphonic acid. The amidine is preferably used as the salt, suitably the hydrochloride salt, which may then be converted into the free amidine in situ by employing a two phase system in which the reactive ester is in an inert organic solvent such as chloroform, and the salt is in an aqueous phase to which a solution of an aqueous base is slowly added, in dimolar amount, with vigorous stirnng. Suitable amidines may be obtained by standard methods, see for instance, Garigipati R, Tetrahedron Letters, 190, 31, 1989.
Compounds of Formula may also be prepared by a process which comprises reacting a compound of Formula wherein TI is hydrogen, with an N-acyl heteroaryl salt, according to the method disclosed in US patent 4,803,279. US patent 21.
4,719,218 and US patent 5,002.942, to give an intermediate in which the heteroaryl ring is attached to the iniidazole nucleus and is present as a 1 ,-dihydro derivative thereof, which intermediate may then be subjected to oxidative-deacylation conditions (Scheme 11). The heteroaiyl salt, for instance a pyridiniumn salt, may be either preformed or, more preferably, prepared in situ by adding a substituted carbonyl halide (such as an acyl halide, an aroyl halide. an arylalkyl haloformate ester, or, preferably, an alkyl haloforruate ester, such as acetyl bromide, benzoylchloride, benzyl chioroformate. or, preferably, ethyl chioroformate) to a solution of the compound of Formula (IX) in the heteroaryl compound R 1 H or in an inert solvent such as methylene chloride to which the heteroaryl compound has been added. Suitable deacylating and :oxidising. conditions are described in U.S. Patent Nos. 4,803,279, 4,719,218 and *'**5,002,942, which references are hereby incorporated by reference in their entirety.
Suitable oxidizing systems include sulfur in an inert solvent or solvent mixture, such as decalin, decalin and diglyme, p-cymene, xylene or mesitylene, under reflux conditions, or, preferably, potassium t-butoxide in t-butanol with dry air or oxygen.
0 P2 HN Pt NRe
R
4 Rt R 4 R Re In a further process, illustrated in Scheme M[ below, compounds of Formula (I) may be prepared by treating a compound of Formula thermally or with the aid of a cyclising agent such as phosphorus oxychioride or phosphorus pentachioride (see Engel and Steglich, Liebigs Ann Chem, 1978, 1916 and Strzybny et al., J Org Chem, 1963, 28, 338 Compounds of Formula may be obtained, for instance, by acylating the corresponding a-keto-ainine with an activated formate, derivative such as the corresponding anhydride, under standard acylating conditions followed- by formation of the imine with R 2
NH
2 The aminoketone may be derived from the Parent ketone by oxamination and reduction and the requisite ketone May in turn be prepared by decarboxylation of the beta-ketoester obtained fromn the condensation of an aryl (heteroaryl) acetic ester with the RICOX componenL 22 X NOOMe O R 1.)NaNO, HO A me N 0 POC 3 4 R4 Mc4
FIBRA
4 Ri NH 2 2.)NyIIA.7~~R .I 4,o frm (X) SCHEME m In Scheme IV illustrated below, two different routes which use ketone (fonrmula XI) for preparing a compound of Formnula A heterocyclic ketone (XI) is prepared by adding the anion of the alkyl heterocycle such as 4 -methyl-quinoline (prepared by treatment thereof with an alkyl lithium, such as n-butyl lithium) to an Nalkyl-O-alkoxybenzamide, ester, or any other suitably activated derivative of the same 10 oxidation state. Alternatively, the anion may be condensed with a benzaldehyde, to give an alcohol which is then oxidised to the ketone 3 RI NHR 2 R, R X,0 0
R
4
R
4 H I F .x o o Y~ NH X1 R1.cLu.B
R
S...R
X RrCH. 0 BtN SCHEME IV In a further process, N-substituted compounds of Formula may be prepared by treating the anion of an amide of Formula (XII): RICH2NR2COH
(XII)
wherein RI and R2 with: a nitrile of the Formula (XIII): R4CN (XII) wherein R4 is as hereinbefore defined, or an excess of an acyl halide, for instance an acyl chloride, of the Formula
(XIV):
R4COHal (XIV) 23 wherein R4 is as hereinbefore defined and Hal is halogen, or a corresponding anhydride, to give a bis-acylated intermediate which is then treated with a source of ammonia, such as amimonium acetate.
R
2 HN base Ri,,C R1.) U+ -N(-i 2 RI N2 0
R~
One variation of this approach is illustrated in Scheme V above. A primary amine (R2NH2) is treated with a halomethyl heterocycle of Formula RiCH 2 X to give the secondary amnine which is then converted to the amide by standard techniques.
Alternatively the amuide may be prepared as illustrated in scheme V by alkylation of the formamide with RjCH2X. Deprotonation of this amide with a strong amide base, such as lithium di-Ws-propyl amnide or sodium bs-(zrimethylsilyl)arnjde, followed by addition of an excess of an aroyl chloride yields the bis-acylated compound which is then closed to an imidazole compound of Formula by heating in acetic acid containing ammonium acetate. Alternatively, the anion of the amide may be reacted with a substituted aryl nitrile to produce the imidazole of Formula directly.
Suitable protecting groups for use with hydroxyl groups and the imidazole nitrogen are well known in the art and described in many references. for instance, Protecting Groups in Organic Synthesis, Gen~e T W, Wiley-Interscience, New York, 1981. Suitable examples of hydroxyl protecting groups include silyl ethers, such as tbutyldimethyl or t-butyldiphenyl, and alkyl. ethiers, such as methyl connected by an alkyl chain of variable link. (CRioR~oM. Suitable examples of imidazole nitrogen protecting groups include terahydropyranyL- Pharmaceutically acid addition salts of compounds of Formula may be obtained in known manner, for example by treatment thereof with an appropriate amount of acid in the presence of a suitable solvent.
In the Examples, all temperatures are in degrees Centigrade Mass spectra were performed upon a VG Zab maws spectrometer using fast atom bombardment, unless otherwise indicated. IH-NMR (hereinafter -NMR-) spectra were recorded at 250 MHz using a Bruker AM 250 or Am 400 spectrometer. Multiplicities indicated are: s=singlet, d=doublet, t~triplet, q=quartet, m=rnulfiplet and br indicates a broad 24.
signal. Sat. indicates a saturated solution, eq indicates the proportion of a molar equivalent of reagent relative to the principal reactant.
Flash chromatography is run over Merck Silica gel 60 (230 400 mesh).
SYNTHETIC EXAMPLES The invention will now be described by reference to the following examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention. ADl temperatures are given in degrees centigrade, all solvents are highest available purity and all reactions run under anydrous conditions in an argon atmosphere unless otherwise indicated.
I-f3-(4-Moroholinvnpropvl1-4-44-fluorophenl-S-(4-pvidvn~inidazole a) 4-fl uoroplhenyl-tolylthiomethvlformaniide A solution of p-fluorobcnzaldehyde (13.1. miiiters (hereinaftermL), 122 mailimoles (hereinafter mmol) thiocresol (16.64 grams (hereinafter 122 mmol), formamide (15.0 reL, 445 mmol), and toluene (300 mL) were combined and heated to toluene reflux witha aotropic removal of H20 for 18 h. The cooled reaction was diluted with EtOAc (500 mL) and washed with satd aq Na2CO3(3 x 100 mL), satd aq NaCI (100 mL), dried (Na2SO4), and concentrated. The residue was trituated with petroleum ether, filtered and dried in vacuo to afford 28.50 g of the title compound as a white solid (85 melting point (hereinafter mp) 119 1200.
T1he compound of example 1(a) (25 g, 91 mmol) in CH2Cl2 (300 reL) was cooled to -300 and with mechanical stirring POC13 (11 mL, 110 mmcl) was added dropwise followed by the dropwise addition of Et3N (45 mL, 320 mmol) with the temperature maintained below -300. Stirred at -300 for 30 min and 50 for 2 h, diluted with CH2CI2 (300 mL) and washed with 5% aq Na2CO3 (3 x 100 mL), dried (Na2SQ4) and concentrated to 500 mL This solution was filtered through a 12 x 16 cm cylinder of silica in a large sintered glass funnel with CH2Q2 to afford 12.5 g of purified isonitrile as a light brown, waxy solid. IR (CH2CI2) 2130 cm* 1 C) ddn -croadhf -M hoiyW 3y* Pyridine-4-carboxaldehyde (2.14 g, 20 tumoL), 4-(3-aminopropyl~morpholine (2.88 g, 20 mmol), toluene (50 mL) and MgSO4 (2 g) were combined and stirred under argon for 18 h. T"he MgSO4 was filtered off and the filtrate was conmintrated and the residue was reconcentrated from CH2C12 to afford 4.52 g of the title compound as a yellow oil containing less than 5% of aldehyde based on IH NMR. IH NMR (CD3Cl): d 8.69 J 4.5 Hz, 2H), 8.28 LH), 7.58 J 4.5 Hz, 2H), 3.84 (in, 6H), 2.44 (mn, 6H), 1.91 (mn, 2H).
d) I -[3-(4-MorpholinylbpropyJ1-4-(4-fliuoropheny[)-5-(4-pyridvfliMid&oIe The compound of example I1(b) (1.41 g, 5.5 mmol), and the comnpound of example I1(c) 17 g, 5.0 mmol) and CH2CI2 (10 mL) were cooled to 5 OC. 1.5.7uiazabicyclo(4.4.0]dec-5-ene, henceforth referred to as TBD, (0.71 g 5.0 nimol) was added and the reaction was kept at 5 0 C for 16 h, diluted with EtOAc (80 mQL and washed with satd aq Na2CO3 (2 x 15 niL). The EtOAc was extracted with I N HC1 (3 x 15 mQL, and the acid phases were washed with EtOAc (2 x 25 niL), layered with EtOAc (25 mQL and made basic by the addition of solid K2C03 til pH 8.0 and then NaOH til pH 10. The phases were separated and the aq was extracted with additional EtOAc (3 x 25 mQL. The extracts were dried (K2C03) concentrated and the residue was crystalized from acetone/hexane to afford 0.94 g of the title compound. mp 149 150 0.
Eample2 1 -(3-Chloronropvfl-4-(4-fluorophenfl)-5-(4-nvidvl)irnidazole To 3-chioropropylazuine HCI (15.1 g, 0.120 moles (hereinafter mol)) and H120 (100 mQL was added pyridine-4-carboxalclehyde (9.55 ml, 0. 100 mol), then K2C03 (8.28 g, 0.060 mol) then CH2CL2 (100 niL) and the mixture was stirred for 40 min.
The phases were separated and the aq phase was extracted with additional CH2C12 (2 x mL), dried (Na2SO4) and concentrated to afford 17.1 g IH NMR (CD3Cl): d 8.69 J 4.5 Hz, 2H, 8.32 1H1), 8.28 1H), 7.58 J =4.5 Hz, 211), 3.71 (mn, 2H1), 3.63 J 6Hz, 2H), 2.24 J 6Hz, 214). The presence of 9% of the aldehyde was evident by IH NMR.
b) 1 /3-Chlo A (fA mlhn l)54-A dl~mda The compound of example 1 (6.85 g, 26.6 mmol), the compound of example 2(a) (6.32g, 34.6 nimol), CH2CI2 (70 mQL, and TBD (4.07 g, 28.4 nimol) were reacted by the procedure of example 1(d) to afford 3.19 g mp 139 1400.
30Exml3 I -(3-AzidopipyI)--(4-fluoropheny)-5-(4-vridy'~imidazole To a solution of the compound of example 2(b) (250 millgram (hereinafter mig), 0.79 nimol) and DMF (5 mQL was added NaN3 (256 mig, 3.95 mmol) and Nal (12 mg, 0.08 mmol) and the mixture was heated to 900 till the reaction was completed based on tic, analysis (19:1 CH2Cl2IMCOH). The cooled reaction was added to 5% sq Na2CO3 (20 niL) and extracted with EtOAc (3 x 25 mL). The combined extracts were washed with H20 (3 x 25 niL and flash chromatographed (2.2 x 10 cm column) with 0 1% MeOH in CH2Cl2 to afford 2.54 mg (100%) of the title compound as a white solid. nip 64 650.
27 Examlpre 9 1 -(3-Diethvlamin opropyi)-4-(4-fl uorophenyl)-5-4-pvridvl)imidazole Following the procedure of example 3 except using diethylamine as the nucleophiie and isolation of the product by extraction with diethyl ether, the title compound was prepared as a white solid (21 yield). mp 94 950.
Examle 1 -Pineridinvl)nropvll-4-(4-fluoronhenvl)-5-(4-vridvl)imidazo Following the procedure of example 3 except using piperidine as the nucleophile and purification of the crude product by trituration with hot he..ane, the title compound was prepared as a white solid (63% yield). mp 105 1080.
Example I 1 -f3-(Meththio)propvI14-(4-fluorophenyfl)-5-4-pdyl)midAZe Following the procedure of example 3 except using sodium thiomethane as the nucleophile and omitting the sodium iodide followed by purification of the crude product by trituration with hot hexane, the title compound was prepared as a white solid (50% yield). mp 85 860.
Example 12 I -r2-(4-Mrholfehi1-4-(4-hiorophenfl(4pridyfimn~ i a. a a a) I;=ij- 7-r IMoMMMT Following the procedure of example 1(c) except using 4-(2aminoethyl)morpholine as the amine, the title compound was prepaed as a light yellow oil (100%) containing less than 10% of aldehyde based on 1 H NMR. IH NMR (CD3CI): d 8.68 J 6 Hz, 2H), 8.28 1H), 7.58 J 6 Hz, 2H), 3.82 2H), 3.72 4H), 2.72 2H), 2.55 4H).
b) 14 2-(4-Moholnylcitl4-4kfuomhenl)--(4-9.yfdllaamd2ZQt Following the procedure of example 1(d) except using the compound of example 20(a) as the imine, afforded the title compound as a white solid (21 mp 114-1150.
Example 13 1-f3-(4-Mohoinvl)propyl-4-(3-methylthiopheny-5-(4-pyridimidgk a) N-f3-mthlthiohenl-(t vlthio)methylfrmnid Following the procedure of example 1(a) except using miethylthiobenzaldehyde as the aldehyde, the title compound was prepared as a white solid mp 103 1040.
28.
b) 3-methvlthioDhenvl-(colvlthio)methvuisocyanide Following the procedure of example 1(b) except using the compound of the previous step as the formamide, the title compound was prepared as a light brown oil IR (CH2CI2) 2120 cm -1 c) 1 -f3-(4-Mopolinh npronvl -4-(3-methyvthiohenvl)-5-(4-Dridvl)im idazole Following the procedure of example I(d) except using the compound of the previous step as the isoniuile, afforded the title compound as a white solid (31 mp 105- 1060.
Examle 14 1-13-(-I4-M oholjyjnroX114(3-ethlsurinypheyl)5-(-pydyl~~id7Ol The compound of example 13(c) (200 mg, 0.49 mmol) was dissolved in HOAc (4 mL). K2S208 (151 mg, 0.56 mmol) dissolved in H20 was added and the solution was stirred for 16 h, poured into 10 aq NaOH (50 mL) (the resulting solution was pH 10) and extracted with EtOAc (3 x 25 mL). The extracts were dried (K2C03), concentrted and the residual oil crystalized from acetondhexane to afford 87 mg of a white solid. mp 117 1180.
ExamULC 1-r3-(N-methyl-N-benzfl-minonropyll-4.3m-mthA ophenvl.4-(4.
pyriddy1imidazole a) Pvridin-A-&boxaldehyde 3-(N-methyl-N-benzylaminronvl)imine Following the procedure of example 1(c) except using 3(-N-Methyl.Nbenzylazino)propylamine as the amine (UedaT.; hsb'izMkiL; Chem. Pharm. Bull.
1967, 15, 228 237.), the title compound was obtained as a light yelow oil (100%) containing less than 10% of aldehyde based on IH NMR. 1H NMR (CD3C1): d 8.65 J 7 Hz, 2H), 8.21 1H), 7.54 J 4.5 Hz, 2H), 7.52 5H), 3.69 J1= 11 Hz, 2H), 3.48(s, 2H), 2.44 J 11 Hz, 2H), 2.18 3H), 91 2H).
b) I -f3-(N-methl-N-benzvaminopropyi1-4(3-methyithiophenyr.5-(4- Vyjidylimidazole Following the procedure of example I(d) except using the compound of example 13(b) as the isonitrile, and the compound prepared in the previous step as the imine afforded the title compound as a white solid mp 87 880.
Examl6 I-13-(N-Methvl-NebenZylaminopoyll-4-(3-methvlsulfinvlphenyh..544 pyddyffimidazole Following the procedure of example 14 except using the compound of example as the sulfide afforded the title compound as a white solid mp 84 850.
29- Examle 17 I-f4-(MethlthioiphenvI-4-(4-fluorophenv)-5-(4-pdridyl)imidazole a) Eyridine-4-carboxaldehdve (4-methylthiophenylimine Following the procedure of example I(c) except using 4 -(methylthio)aniline as the amine afforded (100%) of a light yellow oil with no detectable amount of aldehyde based on 1 H NMR. IH NMR (CD3CI): d 8.75 J 6 Hz, 2ff), 8.47 IH), 7.74 (d, J 6 Hz, 2H), 7.30 J 8 Hz, 2H), 7.22 J 8 Hz, 2H), 2.52(s, 3H).
b) 1 -f4-(Methvthio)phenv1-4-(4-fluomnhenl)-5(44 dyrnI l Following the procedure of example 1(d) except using the compound of the previous step as the imine afforded the title compound as a white solid mp 172- 1730.
4 S *.L 0 of 15 1 -[4-(Methvlsulfinvl)phenvll-4-(4-fluorophenvl)-5-(4-pvridyl~imidazole Following the procedure of example 14 except using the compound of example 17(b) as the sulfide afforded the title compound as a white solid mp 202 2030.
Example 19 1-r3-M etlthiolphenyl14-44-flugrnphvfl~M-5-44pyi midazole a) Evrddine4-caroxaldehdya (3-methylto vhev iminen Following the procedure of example 1(c) except using 3-(methylthio)aniline as the amine afforded of a light yellow oil with ca 2.5% of aldehyde based on IH
S
S 0 NMR. 'H NMR (CD3CI): d 8.76 (d J 6 Hz,2H), 8.44 lH), 7.74 J =6Hz, 2H), 7.30 J 8 Hz, 2H), 7.34 6.98 (ni, 4H), 2.52(s, 3H).
b) I-l-(mCththiWhenv44-4lM~henor)-5envD-5 Following the procedure of example 1(d) except using the compound of the previous step as the imine afforded the title compound as a white solid mp 155- 1560.
Ex Ult~ I 2f-(MethlsulfinyA1- hevl'-(A4 -fluor iA Following the procedure of example 14 except using the compound of example 19(b) as the sulfide afforded the title compound as a white solid mp 233 2340.
Example 71 1 -r2-(Mehvlthio~ben11 a) Pyridine-4-carboxaldehdye (2-methylthiophenyltimine Following the procedure of example I1(c) except using 2 -(methylthio)anilne as the amine afforded of a light yellow oil with ca 8% of aldehyde based on I NMR. 1 H NMR (CD3CI): d 8.75 J 6 Hz, 2H), 8.41 IH), 7.79 J 6 Hz, 2H), 7.36 7.00 (in, 4H), 2.47(s, 3H1).
b) 1 -r2-(methvlthio)phenyll-4-(4fluoohenX]-5-(4-pvridvlbznidAzole Following the procedure of example I1(d) except using the compound of the previous step as the imine and purification by flash chromatography with 0 -1I% MeOH in CH2CI2 afforded the title compound as a non-crystalline white foam mp =59 60 0 Example 2 -f -rMethvylsulfinvflplhenyl1-4-4-fluorophenvi)-5-(4-nvridvl~imidazole is Following the procedure of example 14 except using the compound of example 2 1(b) as the sulfide, and purification by flash chromatography with 0 MeOH in CH2C12 afforded the title compound as a non-crystalline white foam mp 60 0:461650. (M'e ill defied mp is probably the result of a mixture of conformational isomers 00#6 which is clearly indicated in the IH and 13 C NMR spectra of this compound.) we. -(-hgM I4(-l~ezainvJ4-M&Di.Zao (See also Example 2 above for alternative method of preparation) a) 4 FlMheynnymcflommd To a solution of toluene sulpbinic acid sodium salt hydrate (120g) in water (750 ml) was added concentrate sulphuric; acid (16m1). Dichioromethane (5001) was S. added and the organic and aqueous layers were sepaazed the aqueous layers being extracted with dichloromethan (2x20m1). The Combined organic extracts were dried (Na2SO4) and evaporated to dryness to yield the solid suiphinic acid (71.79g, 0.46 mole). This was added to p-fluorobenzaldehyde (57.04g, 0.46 mole) and formamide (62. 1g, 1.38 mole) and the resulting mixtur was stined with camphor-LO0-sulphonic acid (21.3g, 0.092 mole) at 60-65", under nitrogen for 22 hours. A solution of sodium bicarbonate (33.6g, 0.40 mole) in water (400rnl) was added to the ice-cooled solid product which was broken up and stirred for 30 minutes. The crude product was collected and washed with acetone (220m1l) and then ether (3x220i) and dried to yield the desired product, 91.5g, 64.8%.
b) 4-loohM-=Iehlsond To a suspension of the compound of the previous step (3.22g. 10.5 minole) in dimethoxyethane (21m1) stirring at 100 was added phosphorus oxychloride (2.36ml, 31.
25.3 mmole) dropwise over 5 minutes. Tniethylaniine (7.35m1, 52.5 mmole) was then added dropwise over 10 minutes and the reaction mixtur. was poured into saturated sodium bicarbonate solution (lO0rnI) and the oily product was extracted into dichioromethane (2x3ml). The organic extracts were evaporated to yield a black oil (3.5 1Ig) which was eluted from Grade MU basic alumina (60g) using dichloromethane.
The combined product fractions were evaporated and ether added to crystallize the desired product, 1.735g, 57%.
C) I -(3-Chloropyfl1-4-(4-fluoronhenl)-5-(4-nvridfl)-imidazole To a solution of the compound of the previous step 183g, 4.O9mmol) and the compound of example 2(a) 122g, 6. l5mmole) in dimethoxyethane (I 5mi) at ambient temperature wits added dropwise, over 10 minutes a solution of DBU (0.67im1, in dimethoxyethane (l1rel). The reaction mixture was stirred at ambient temperature for 1-1/2hours and then evaporated to leave an oil which was eluted from Grade il bisic alumina (100g) to yield the desired product, 1.096g, I -r4-(4-MoMholinyj)butyll-4-(4-fuomphenl)-5-(4-pyidflimida.zcle a) 4-(4-Morpholinombutyl- I-pthalimide 4-Bromobutyl-lI-pthalimide (5.0 g, 17.7 mmol) and morpholine (20 niL) were combined and stirred for 3 h, diuted with Et2O (200 niL), and filtered. The solid was washed with more Et2O and the combined filtrates were extracted wih 3 NHCl(3 x niL). The combined acid phases were washed with Et2O (3 x 5Om),Ilayered with EtOAc andi made basic by the addition of solid K2C03 til the foaming stopped and then 10% aq NaOH was added Wi the pH was 10. Extraction with EtOAc: (3x. 100 niL), drying, (K 2 C03), concentraion and flash filtration I L silica wash first with 0 -4% MeOH in CH2C12 and then elute product with 4% MeOH and 1% Et3N in CH2Cl 2 to afford 5.52 g of the title compound as a white sc-.A.
b) 4-4-gwkh~no~u~an~ T'he compound of example 24 (1.0 g, 3.47 mmol), hydrazine monohydrate (190 pl. 3.82 nimol) and CH 3 0H (20 niL) were combined and stirred at 230 overnight.
The CH 3 0H was removed in vacuo and the resudue was concentrated to dryness from EtOH. The residue was combined with 2N HQI (20 niL) and stirred for 2h, filtered and the solid was washed with H20. The combined filtrates were concentrated in vacuo and reconcentrated from EtOH twice to give a white foam which was dissolved in 3:1 CH2ClICH 3 OH, and stirred with solid K 2 C0 3 for 5 min and filtered. The filtrate was concentrated to afford 0.535 g of a brown oil. 1 H NMR (CD3Cl): 3.7 -3.2(in, 6H), 2.7 2.2 (mn, 1.6 1.3 (in, 6H).
C) Pyridine-4-carboxaldehvde [4-(4-morphglinflbutvllimine 32..
.Following the procedure of example I1(c) except using the compound of example 24(b) as the amine the title compound was prepared as a light yellow oil (100%) con taining 30% of aldehyde based on I H NMR. I1H NMR (CD3Cl): 8.60 J 6 Hz, 2H), 8.19 I 7.51 J 6 Hz. 2H), 3.7 -3.2 (mn, 6H), 2.5 2.2 (in, 1.7 1.4 (mn, 4H).
d) I -f4-(4-Morp~holinyl)butyl 1-4-(4-fluorophenyl)-5-(4pyndvl)imidazoQle Following the procedure of example I1(d) except using the compound of example 24(c) as the imine afforded the title compound mp 103 1040.
1 -Cyclopropyl-4-(4-fluorophenyfl-5-(4-pyridyflimidazole a) Eydn--abxleye=o lmin Following the procedure of example 1 except using a 100% excess of the volatile cyclopropylanune the title compound was prepared as a light yellow oil 1 fl NMR (CD3Cl): 8.65 J 6 Hz, 2H), 8.40 1H), 7.51 J 6 Hz, 152H), 3.07 (mn, 18), 1.01 (mn, 4H) b) I -Cvcloproolyl-4-(4-fluorophenfl)-5-(4-pyrddvIimidazole The compound of the previous step (20 minol), the compound of example 1 (b) (5.65g, 22 inmol), and CH 2 Cl 2 (20 ml) were cooled to 00 and TBD (2.84g, 20 minol) was added. Stirred at 50 for 2h, 230 for 48h and refluxed for 4 h. The crude reaction was flash filtered through a sintered glass funnel filled with silica (I L of silica) eluting with 0 CH30H in CH 2 C12. Crystals from hexane/acetone to afford 839 mg mp =l129.0 -129.50.
1 -Isopropvl4-(4-fluoohenyh-5-(4-yidvlimidazole a) E~ik_-abxleX s=21mn aieFollowing the procedure of example I1(c) except using isopropylainine as the *aie the title compound was prepared as a light yellow oil IH NMR (CD3C1): 8.67 I 4.4 Hz, 2H), 8.27 IH), 7.59 J 4.43 Hz, 28), 3.59 (in, 1lH), 1.o27 J 6.3 Hz, 68).
b) I -IsoprOpyl4-(4-fluorophenyll)-5-44-p.Yrdv1)iMidazole Following the procedure of example 1(d) and substituting the imine of the previous step the compound was prepared using a modified work up of flash filtration of the crude reaction through silica (0 CH 3 0H in CH2C12). Two crystallizations from hexane, /acetone afforded the title compound as tan needles (30 mp 179.0 179.5.
Exampl27
M
a) Eyridine-4-carboxaldehyde Cyclopropvlmethvlimi'_ Following the procedure of example I1(c) except using cyclopropylmethylane as the amine the title compound was prepared as a light yellow oil 1 H NMR (CD3CI): 8.69 J 4.5 Hz, 2H), 8.27 JH), 7.61 J 4.5 Hz, 3.55 (di, J 6.7 Hz, 2H), 1. 15 IH), 0.57 (mn, 2H), 0.27 (mn, 2H).
b) I -Cyclopropylnlethvl-44-fluoroohenfl-5-44-D~ridyl)iinidazole Following the procedure of example I1(d) substituting the iinine of the previous step the compound was prepared using a modified work up of flash filtration of the crude reaction through silica (0 CH30H in CH 2 C1 2 Crystallization from hexane /acetone afforded the title compound as white flakes (62 mp 162.0 162.5.
I -tert-Butvl-4-(4-fluoinphenfl)-5-(4-pvndylliiidazole a) Eydn--abxJeyetr-umn Following the procedure of example I1(c) except using tert-buryLarnine as the.
amine the tidle compound was prepared as a light yellow oil IH NMR (CD3C1): 8.67 J 4.4 Hz., 2H), 8.22 1H), 7.61 (di, J 4.4 Hz, 2H), 1.30 9H).
b) I -tert-ButyI-4-4-fluonphcyl)-5-(4-plyrdflimidazole Following the procedure of example 1 substituting the imine of the previous step the compound was prepared using a modified work up of flash filtration of the crude reaction through silica (0 CH30H in CH 2
CI
2 to afford the title compound as tan powder (16 mp 199.0 200.0.
1 -(2.2-Diethoxvethvfl)4-(4-fluomophenylI.5-(4-pvridyflimidatzole a) 167M A~ P TT ~M7 .Following the procedure of example 1(c) except using 2,2-diethoxyethylamnine as the amine the tidle compound was prepared as a light yellow oil 1 H NMR (CD3C1): 8.69 J 4.4 Hz, 2H), 8.28 I 7.60 J 4.4 Hz, 2H), 4.82 J 5.1 Hz, IHO3.82 J =5.1 Hz, IH), 3.72 3.57 211), 1.21 I 7.3 Hz, 6H).
b) I -(2.2-Dietxvethl) A -(4-fluorophenl- 5-(4-pyfidyimidao Following the procedure of example I1(d) substituting the imine of the previous step, the compound was prepared using a modified work up of flash filtration of the crude reaction through silica (0 CH30H in CHI2l2), followed by a flash chromatography through silica with 25 100% ROMc in hexane) and trituration of the resulting gum with hexane afforded the title compound as a white powder (47 mp 69.5 70.0.
34., 1 -Foamvlmethyl-4-(4-fluorophenvl )-5-(4-nvridvh)imidazple The product of example 29(b) (400 mg, 1. 13 mmol), H 2 0 (10 MiL), acetone mnL) and concd H2S0 4 (1 niL) were combined and refluxed for 24 h. Most of the acetone was removed in vacuo and the residue was combined with 5% aq Na2CO 3 and extracted with EtOAc, dried (Na2SO 4 concentrated and crystallyzed from acetone to afford the title compound as a white powder (47 nip 118.5 119.0.
Exam~l 1 The product of example 30 (317 mig, 1. 13 minol), hydroxylamine hydrochloride (317 mug), pyridine (317 IiL), and EtOH (3.8 niL) were combined and refluxed for 3h, poured into 5% aq Na 2
CO
3 and extracted with EtOAc, dried (Na2SO 4 and flash filtered in 0 4% MeOH in CH2CR2 to afford 261 mug of the title compound as a :15 white powder. nip 184.0 185.0.
I-Cvanoniethl-444.fluorophenXyw5-4-pyvimidazole The product of example 31 (250 mg, 0.84 minol), and CuSO4 were combined and refluxed for 2h. The cooled reaction was flash filtered in 0 MeOH in CH 2
CI
2 to afford 129 mng of the title compound as a white powder. mp 132.0 133.0.
I 3 4 -Monhoinylpropyl14-(4-fluoroplhenyl)-5-(2metypyid4vliumdaze *25 a) 4-Fonnyl-2-meth3Epyridin'e 4-Cyano-2-inethylpyridine was prepared from 2,6-lutidine asccording to the literature procedure (YarnanakaRH; AbeH.; Sakanot-iT.; Hidetoshi, HiranumaH.; KamataA Chem. Pham. Bull. 25(7). 1821 1826.). A solution of 4-Cyano-2niethylpyridine (0.367 g, 3.11 inioL) and toluene (3.5 rnL) was cooled to -780 and 1 M DIBAL in hexanes (3.6 inL, 3.6 nimoL.) was added dropwise via syringe pump (T -650). The reaction was warmed to 50 and stifred for 5 mini, recooded to -780 and CH3H (3.5 ml)waed< wa ed t0 nd sdfor5 mnand thn aq Rochelle's salt was added, stirred for 3 min and then acidified to pH 1.0 with aq H2S0 4 The aqueous was made basic by the addition of solid K2C03 and extracted with EtOAc. The extracts were dried (Na2SO4) concenated and filtered thijugh silica MeOH in CH 2 Cl 2 to afford 253 mg of aldehyde. HI NMR (CD3CI): d 10.05 1H1), 8.74 J 7Hz, I 7.51 LH), 7.30 J 7Hz. 111), 2.68 3H1).
b) Ecdn4crojl~je[4-o~oiypoylmn
M
The product of the previous step and 4 3 -axinopronyl)morpholine were reacted by the procedure of example I(c) to afford the title compound as a yellow oil containing no aldehyde based on 1 H NMR. I H NMR (CD3CI): d 8.57 J 5.0 Hz, IH), 8.25 7.46 IH); 7.36 J 5.0 Hz, IH), 3.71 6H), 2.60 H); 2.35 6H), 1.90 2H).
c) 1-r 3 4 M orholinvnpRrovI)-4-(4-fluorohnyl)-5-(2-methyIMXdy)imidazole The compound of the previous step, and the compound of example 1(b) were reacted by the procedure of example 1(d) to afford the title compound as a white solid [5I from mp 116 1170 ExamIZle 34 4 44-Fluoroahenyfl. I-r 3 4 mopholinyI)p2rOpyjl.(2-lcroddinyl) imidazole a) 2:Chlorovrldine.4carboxajI ehyde 0344morpholinvilproPvllimine 2-Chloropyridine4-carboxaldehyde was prepared as described in the patent literature (WPI Acc. No. 88-258820/37) whose disclosure is incorporated by reference in its entirety herein. This aldehyde was reacted with 4-(3-aminopropyl)zorpholine by the procedure of example 1(c) to afford the title compound as a yellow oil. IH NMR (CD3CI): 8 8.45 J 5.1 Hz, 18), 8.24 1H), 7.63 1H); 7.51 J 5.1 Hz, IH), 3.72 61), 2.44 6H), 1.91 21).
b) 4-(4-Fluahenyll 1- rM-14mornhoinllpmgyll-5--2-chhm-4-pyddinyd Following the procedure of example I(d) except using the compound of the pre- 0 vious step as the imine afforded the title compound as a white solid mp 97.0- 97.50.
4 4 -PluoRoheny)- I -r3-(4-morholinyl)propyll -5-(2-amin-4-nRyrdiny)l imidazole a) A (luornhenyl A fL.3-(4-moIAolinvlUwowl (-hIAvjn; 1 Ani a imidazole The compound of example 34(b) (872 mg. 2.18 mmoL) and 98% hydrazine hydrate (9 mL) was heated to 1150 (bath terup) for 20 h, cooled to 230 combined with
H
2 0 (20 inL) and extracted with EtOAc (3 x 25 mL). The combined extracts were washed with H20 (2 x 20 mL) and dried (Na2SO 4 Flash chromatography with 0 8%
CH
3 0H in CH 2 C1 2 afforded 547 mg (63) the title compound as a white solid.
b) 4(4-Fluorophenyl)- I -f 3 4 -morpholinvl)propyl-S(2-amino-4-pwinvI~ imidazole The compound of the previous step (100 mg, 0.25 mmoL), absolute EtOH i) and Raney Ni (0.4 mL) were hakrnunder H2 (45 psi) for 4 h. Flash 36 chromatography with 0-8% CH 3 0H in C-H2CI2 afforded 34 mg the title compound asa white solid. mp 186 1870.
Pyiidine-4-carboxaldehyde was reacted with methyl-4-aininobutyjzte by the procedure of example 1(c) to afford the title compound as a yellow oil. IH NM (CD3CI): 6 8.69 J 5.8 Hz, 211), 8.27 1H); 7.56 J 5.8 Hz, 2H), 3.70. (mn, 2H); 2.31 J 8.0 Hz, 211), 2.08 (mn, 2H).
b) I -3-Methoxycarbonyl- I -propyl)-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole Following the procedure of example 1 except using the compound of the previous step as the imine afforded the title compound as a white solid nip= 0 00 69.0 70.00.
Ex&aml 1 I -(3-Carboxy- 1 -propyl)-4-(4-fluorophenyl)-5-(4-pyridyl)imidazole The compound of example 36 (100 mg, 0.29 minol), CH30H (3 inL), and THF (1.5 niL) were combined and the resulting samn was trated with a soin of LiOH (62 20 mug, 1.5 mmiol) in H 2 0 (1.5 rut) and the resulting soin was stirred for 4 h. Removal of the volatile compounents in vacu, redisolving in 1120 and croatography through 0. e* HP-20 with H20 til the eluates were neutral and then with 25% aq MeOH afforded the 000 0title compound as the lithium salt; 65mg ES MS We/ 326 EUDki39 Pyridine-4-carboxaldehyde was reacted with fl-alanine methyl ester by the procedure of example 1 to afford the title compound as a yeliow oil. IH NMR (CD3C1): 8 8.68 J 4.5 Hz, 211), 8.33 1H); 7.57 J 4.5 Hz, 211), 3.93 J= 6.7 Hz, 21H); 3.69 3H1), 2.76 J =6.7 Hz, 211), b) S J I~ l 1930 Following the procedure of example I1(d) except using the compound of the previouis step as the imine afforded the title compound as a white solid (40% from the amine). up, 119.0 120.00.
FAMS~l 3 I -(2-Carboxy- -ethyl)-4-(4-fluoropheiyI)-5-(4-pyridyl)imidazole 37 The compound of example 38(b) was nydrolysed by the procedure of example 37 to afford the title compound as the lithium salt; ES MS m/e 312
(MH)
Frample 1-Cl -Benrjzvl peridm4vyl 4 4 -fluorophenyl)-5-(4-2Wi) iMidiazole a) Pvndine=4-carboxadehde (1-benzyridjn4.yJ)jmjn Pyridine-4-carboxaldehyde was reacted with 4 -ainno-N-benzylpiperidine by the procedure of example I(c) to afford the title compound as a yellow oil.
b) I-(I-Benzpl ip~idin-.v)l.(4fluorohen 1)5( 4 4 ilidazOle Following the procedure of example 1(d) except using the compound of example the previous step as the imine afforded the title compound as a white solid from the amine). ES MS mle 413 Examle 41 2 -Aminoprimidin-4..vl..4 (4lfluoronheny) -13-I4-Moohoin imidazole a) 2 -Aminy~rimidine-4-carboxal dehyde dimethyl acetal Dimethylformamide dimethyl acetal (55 m, 0.41 mol), and pyruvic aldehyde dimethyl acetal (50 nL, 0.41 mol) were combined and heated to 1000 for 18 h. Methanol was removed in vacuo to afford an oil.
:A solution of NaOH (18 g, 0.45 mol) in H20 (50 mi, was added to guanidine HO (43 g, 0.45 mol) in H 2 0 (100 ml), and the resulting solution was added to the above described oil. The resulting mixture was stirred at 230 for 48 h. Filtration afforded 25g of the title compound.
b) 2-Amino~yrmidi ncarboxxadehde The compund of the previous step (1.69 g, 10 mmol) and 3N HCI (7.3 SL, 22 mmol) were combined and heated to 480 for 14h, cooled, layered with EtOAc (50 miL) and neutralized by the addition of NaHCO 3 (2.1g. 25 mmol) in small portions. The aq phase was extracted with EtOAc (5 x 50 mL) and the extracts were dried (Na2SO4)and concentrated to afford 0.793 g of the title compound.
C) 2 -Aminonvrilidine-4-carboxaldehvde 3 4 -Momholiyl)DRDelylimine The compound of the previous step and 4 3 -amninopropyl)morpholinewere reacted by the procedure of example 1(c) to afford the title compound as a yellow oil.
d)
S-(
2 -Aminovrimidina-vl-4-4fluoophenv 14344 Moholinvrpronvi imidazole SB 216385 38.
Following the procedure of example I1(d) except using the compound of the previous step as the imine afforded the title compound as a white solid. 1
H
NMR (CD3Cl) 5 8.15(d, J 5.4 Hz, I 7,62(s, IH), 7.46 (dd, 7.00~(t, J 8.6Hz, 2H), 6..50(d, J 5.4 Hz, I 5.09(brd.s, 2H), 4.34(t, J =7.0 Hz, 2H), 3.69(m, 4H), 2.35(brd.s,4H), 2.24(t, J 4.6 Hz, 1.84(m, 2H).
ExampleA2 5-(2-Aminoovrimidin-4-fl)-4-(4-fluornphenfl)- 1-(1 -benflglipridi-4-Vflimidazgle a) 2-Aminogdvrmidine-4-carbpxaldehvde (I -benzylziperidin4-v~imille 2-Arninopyrimidine-4-carboxaldehyde and 4 -arninobenzylpiperidine were reacted by the procedure of example I1(c) to afford the title compound as a yellow oil.
b) 5S-(2-Aminonvrimidin-4-vfl--(4-fluorophenfl)- 1-(1 -benZylipridin4 yximidazole Following the procedure of example 1 except using the compound of the previous step as the imine, afforded the title compound as a white solid (31 from the amine). mp 227 -2290 (dec).
Exml 434 5-(2-AlninopyriMidin-4vfl-4-(4-fluorophenfl)- I-(2-propyflimidazole a) 2-AxninopvriMidine-4-carboxaldehy-de (2-proylimine 2 -Aminopyrimidine-4-carboxaldehycie and 2-propyl amine were reacted by the procedure of example I1(c) to afford the title compound as a yellow oil b) 5-(2-Aminopvrmidin-4-yfl-4-(4-fluorophenfl)- I-(2-prpyflimidazole Following the procedure of example 1 except using the compound of the previous step as the imine afforded the title compound as a white solid (32% from the 2-aminopyrimidine aldehyde). mp 201 2020.
Exml 440 2-Aininopyrimidine-4-carboxaldehyde and 2 -cyclopropylmethyl amine were reacted by the procedure of example I1(c) to afford the title compound as a yellow oil.
b) 5i-(2-Anminopyrimidin-4-yfl-4-(4-fluorophenvn)- 1 -(cycglopropvl- MethDiij1A7&j Foilowing the procedure of example 1 except using the compound of the previous step as the inuine afforded the title compound as a white solid (38% from the 2-aminopyrimidine aldehyde). mp 187 1880.
39 Example 5-(2-Aminopvrimidin-4-yfl-4-(4-fluoronhenvl)- I -carbo~yethyl-4piperidinyl)irnidazole a) 2-Aminopvrimidine-4-carboxaldehyde (I -carboxvetyl-4-UiperDinvlmine 2-Aminopyrimidine-4carboxaldehyde and I -carboxyezhyl-4-aminopiperidine were reacted by the procedure of example I1(c) to afford the tidle compound as a yellow oil.
b) 5-42-Aminopvrimidin-4-vfl-4-4-fluorophenvl)-lI-(I -carboxvethyvl.4- Following the procedure of example I1(d) except using the compound of the previous step as the imine afforded the title compound as a white solid (26% from the 2-axuinopyrimidine aldehyde). mp 216 2180 (dec).
5-(2-Axninonvriinidin-4-vfl-4-(4-fluorophenyfl)- I-(4--2ineridinvfl imidazole :0 a) 2-Aminopyrimidine-4-carboxaldehyde (I -t-butoxycarbonl-4amnp*9*njimn 2-Aminopyrimidine-4-carboxaldehyde prepared in Example 41 and l-tbutoxycarbonyl-4-aminopipenidine (Mach R. etal.,LJ dd Chelm., 36, p3707- 3719, 1993) were reacted by the procedure of example 1 to afford the title compound as a yellow oil.
a klmlQZy.aLk.Qlyljdin~KiDlf iiidazole Following the procedure of example I(d) except using the compound of the previous step as the imine afforded the title compound as a white solid (27% :09:from the 2-aminopynmidine aldehyde).
imidazole SB-220025 The compound of the previous step was combined with 4N HCI in dioxane (5 mL), stirred 10 min, diluted wilth EtOAc and the liquid phase was decanted. The solid was twice washed with Et 2 O (25 mL) and the liquid phase was decanted. Further trituration with EtOH (abs) and then Et: 2 O and drying in vacuo at 500 for 16 h afforded the title compound as the trihydrochioride mp 265 275 (dec) Example 4 I -Mety1-4-phen-5-(4-nvridvimidrzle Following the procedure of example 48(b) except using benzonitrile the title compound as a white solid. mp 161-1620.
Example 48 1 -Methvyl-4-r3-(chlorophenvl)1-5-r4-pyDidinYllimidazole a) N-(4-Pvridinvlmethvl)-N'-methvlformamide To a stirring, argon-purged solution of 4-picolyl chloride-HCI (15g. 91.4 mmol) and N-methylformamide (53.4 mi, 914 mmol) in 300 mi of THF at room temperature was added portionwise over a 20 minute period a suspension of 80% NaH (5.48g, 183mmol). The reaction was quenched 18h later by the addition of ice, partitioned between methylene chloride and water, washed with water and brine, dried over MgSO4, and evaporated to dryness to afford a dark oil. Flash chromatography on silica gel provided 10.5g of the titled compounds a pale yellow oil. TLC; silica gel (9:1 CHCI MeOH) Rf 0.54.
b) I-Methyl-4-f3-(chlorophenvl)l1-5-r4-pyridinvllimidazole To a stirred, argon-covered, -780 solution of lithium diisopropylamide (hereinafter LDA), (prepared from 11.2 ml of diisopropylamine in 150 ml of tetrahydrofuran (hereinafter THF) by the addition of 31.9 ml of 2.5M n-BuLi in hexanes) was added dropwise the product of the previous reaction (10g, 66.5 mmol) in 100 mL of THF. Stirring of the resultant reddish-brown solution was continued at -78o for 40 min at which point 3-chlorobenzonitrile (18.3g, 133 mmol) in 100 mL THF was 20 added dropwise over 20 min. The reaction was allowed to warm to room temperature, stirred for lh and heated to reflux for 12 h. The reaction was cooled and worked up in a manner similar to the previous reaction. Flash chromatography on silica gel provided 2.15g of an oil which was crystallized by dissolution with heating in 10 mL of ethyl acetate. Following crystallization, the solid was collected, washed, and dried (0.4 mm Hg) to afford 1.43g of the titled compound as a light tan solid. mp 119-1210.
Example 49 I -Methyl-4-(3-methylthiophenyl-5-(4-5vridvlimidaz Following the procedure of example 48(b) except using the compound of example 13(b) as the aryl nitrile the title compound was obtained as a white solid. ES m/e= 281 (MH+) Example I-Methyl-4-(3-methylsulfinylphenlh5-(4-pyridvlimidazole Following the procedure of example 14 except using the compound of example 49 as the sulfide the title compound was obtained as a white solid. ES MS m/e 297 (MH+) Example51 (+/-)-4-(4-FRuorophenvf- 1-r3-(methvlsulinvrovll-5-(4-nvridinvflimidazole 41.
Following the procedure of example 14 except using the compouna [rorn example I11 as the sulfide and quenching with saturated aq NH40H afforded the title compound as a white solid (0.87 g, mnp =122 1230.
Example 2 4(4-Pluorophenyfl.. -f( 3 -mrthlsulfonyl)ronvil-5.(4Dvlidinyl)imidazole The compound of Example 51 (0-5087 g, 1.48 nimol) was dissolved in methanol (8 ml) and cooled to 0 0 C. The addition of trifluoroacetic acid 12 ml) was followed by the dropwise addition of meta-chloroperoxybenzoic acid (0.23 g, 2.22 mmol) dissolved in CH2CI2 (10 ml). After stirring for I h the solvents were evaporated. The residue was partitioned betwe en H20 and EtOAc and the aqueous phase was made basic by the addition of 2N NAOH. The organic phase was separated, dried (MgSO4) and concentrated and the residue was pruied by flash chromatography (silica gel. 5% MeOH/CH2CI 2 to afford the title compound (0.37 g, mp =146 15 1470.
To a solution of the compound from example 2(b) (0.22 g, .70 nimol) in acetonitriie (10 ml) was added K2C03 (0.19 g, 1.40 mmol) and phenol 10 g. nimol). After stirring at W0C for 24 h the reaction was diluted with H20. The organic phase was separated and concentrated and the residue was purified by flash chromatography (silica gel 5% MeOHICH2CI2) followed by recrystalization in hexane to afford the title compound (0.02 g, as a white solid. nip 95- 960.
Following the procedure for example 3 except using thiophenol as the nucleophile, adding 2.2 eq K2C03 and omitting the NaL. The cooled reaction was diluted with 10% NaOH and the product was extracted with ether. Flash chromatography was folowed by recrystalization from hexane to afford the title compound 13 g. 53%) as white needles. nip 98- 990.
Quinoyl-4-carboxaldehyde and 4-(3-aminopropyl)mnorphohwn were reacted by the procedure of example I1(c) to afford the title compound as a yellow oil.
b) 1-3 IA k~hoTL X I'I1 A IA4-1. rphll-5(4auiov iid 42 Following the procedure of example 1(d) except using the compound of the previous step as the imine afforded the title compound as a white solid (48% from the amine). mp 139.5 140.0.
Example 56 -(3-Phenvlsulfinvlpropyvl )-4-(4-fluoronhenvl)-5-(4--ovridinvl)imidazole Following the procedure of example 14 except using the compound from example 54 as the sulfide and quenching with saturated aq NH40H afforded the title compound as a white solid. mp 146.5 1480.
Example 57 1 -(3-Ethoxpropyl)-4-(4-fluorohenyl)-5-(4-povridinvllimidazole To a solution of the compound of example 2(b) (0.40 g, 1.26 mmol) in ethanol mi) was added sodium ethoxide (0.8 ml, 21 wt% in ethanol). After refluxing 16 h the mixture was cooled, diluted with H20 and extracted with EtOAc. Concentration of the solvent and purification by flash chromatography (silica gel, 5% MeOHI CH2CI2) afforded the title compound (0.05 g, 12 mp 85 860.
Example 58 20 1-(3-Phenvlsulfonylprovl-4-(4-fluorophenvl-5-(4-pvrdinvfimidazole Following the procedure for example 52 except using the compound from example 56 as the sulfoxide and recrystalizing from hexane following the 9:chromatography afforded the title compound as white solid. mp 109 1100.
Example 59 1 -[3-(4-Morholinvlropvll-4-(3-chlorohenvll-5-(ridvilimidazole a) 3-chlorohenvl-tollthiomethAlisocvanide Following the procedure of example l(ab) except using 3-chlorobenzaldehyde as the aldehyde component the title compound was prepared.
b) 14 3-(I4-MO holinip v ll-4-(3-chlorohl-5-4pyd imidazole Following the procedure of example 1(d) substituting the isocyanide prepared in the previous step the title compound was prepared. MS-DCI NH3 383 Example 1- 3-(4-Mombolinvibronvll-4-(3.4-dichlompbenvl)-54-idvilimida: ole Following the procedure of.example 1(d) substituting the isocyanide prepared in Example 67(a) the title compound was prepared. mp 1060.
43..
4-L4-4-Fluorophenyl)- I -f3-(4-morpholinyD12ropyl 1 H-pyimid-2..OM-Wi yl .01Cazl a) 2-Methvlzhiopvrimidine-carboxuldehyde f3-(4roMinXD~L pXuliiwne Following the procedure of example I1(c) except using 2-methylthiopyrijni.
din-4-crboxaidehyde [Bredereck H. et if. Chem. Ber 1964.,3407] afforded the title compound as a Yellow oiL b) 4- IA u pmy) T:mo~i ")royl-5-(H-R3id2on..4 Concentrated aqueous ammnoniumn hydroxide (2 niL) was Added to 4-(4fluoropheayl)-5-[2-(znethylsulfmnyl)-4pyrimidiny14~3-(4-morphol)propyj..
imidazole (0.14g, 0.37 mazol) [prepared in Example 631 and the reaction mixture was heated to 150 0 for 18 h. After cooling to ambient temperature, the ammoniu hydroxide was decanted. The residue was purified by flash chromatography eluting successively with 4% and 10% methanol in dichiorothane followed by successive elutions with mixtures of 90/10/1l and 7W3W3 chiorofon imethanol/concentrated ammonium hydroxide. Trituradon with ether afforded the title compound as an off-white solid (0.035 g, 24%).
ESMS 384 +H1).
Following the procedure of 1(d) except using 2-methylthiopyrimidine-4- 25 carboxaldehyde (3-(4-morpholinyl)propyljiiine [prepared in Fxaniple 61(a)J afforded the title compound as a yelow, oil. IH NMR (CDCI3) 8 8.31(d, J 7 Hz, 111), 7.64(sIH), 7.46 (dd, 2H1), 7.05(t, J =8 Hz. 2H1), 6.8 1(d. J 5 Hz. 1H). 4.42(t.
J 7.5 Hz. 2H1), 3.7 1(t, I 5Hz, 4H), 2..59(s, 3H), 2.37(brd. s. 4H1), 2.27(t, J 6 Hz, 2H1), 1.85(m, 211).
W~flmica A solution of K2S208 (0.20 g, 0.73 nimol) in water (5 niL) was aidded to fl4-uorophenyl)-5-[2-(methylthio)-4-pyrimidinylj-1l.3-(4lDofpholiy)propylimidazole (0.20 g. 0.48 inmol) in glaia acetic acid (10 niL).
After strng at ambient temperature for 72 b, the reaction nxmitr was poured into water, neutralized with concentrted aqueous ammonium hydroxde and exracted four times with dihooehn. The organic phases were combined 44.
and evaporated. The residue was purified by flash chromatography eluting successively with 4% and 10% methanol in dichioromethane to afford the title compound as a clear oil (0.15 g, IHlNMR (CDC13) 68.57(d, J =7 Hz, 1H1), 7.77(s,IH), 7.47 (dd, 211), 7. 18(d, J 5 Hz, IHl)7.09(t, J 9 Hz, 2H1), 4.56(m, 2H1), 3.72(t, J 5Hz, 4H), 3.00(s, 3H), 2.40(brd. s, 4H), 2.33(t, J 8 Hz, 2H1), 1.94(m, 2H).
Exam I -Propenyfl..4.(4-fluorophenyl-5-(4-Dvridinyflirnidazole a) Exdn--abxleh~ 2Ro y~mn Pyridine-4-carboxaldehyde and 2-prope nyl amine, were reacted by the procedure of example 1 to afford dhe title compound as a yellow oil.
I -Prpenvl- 444-fluorophenfl)-5-(4-pyridinl)irnidazoie o h previous step as the imine afforded a mixture of the title compound and 1-(2- Propenyl)-4-(4-fluorophenyl)-5-(4-pyridinyl)imidazole. Chromatography of the mixture with 0 -50% EtOAc: in hexanes afforded the title compound mp 173.5 -174.0 Eape6 Funthur chromatography of the mixture from example 64(b) afforded the title compound mp 116.0 -117.0 Woo* Example 66 54-(2-N.N-Dimezhytimio)pimidi-4yn4M-flurophcnvI- I -r3-(4-morhoUly- =12ynmidazol Following the procedure of example 61 except using aqueous dimethylamine, afforded the title compound as a yellow glass. ESMS 411 a) 4-trifluoromethylplhenyl-tolylthiomethlsocyanide Following dhe procedure of example I1(ab) except using 4-trifuoromethylbenzaldehyde as the aldehyde, component the tide compound was prepared.
1b) 1 .r3-(4-Morpholinyl)ppv-v54-pUridinyfl-4-r[44trifluommethI~- The imine prepared in Example 1(c) was reacted with the isocyanide prepared in the previous step using the procedure of example 1(d) to prepare the title compound was prepared. mp 1330.
Fxampe 68 1 -f3-(4-Morpholin vhmrofyll-5- (4-nvridinl)4-(3-(trifluoromethvl)phenv11irnidazole a) 3-trifluamomethlhenI-tovlhioethlixsa Following the procedure of example 1(ab) except using 3-trifluoromethylbenzaldehyde as the aldehyde component the title compound was prepared.
b) I -r3-(4-Moholinvl)nropyll-5-(4-odvridinvl)-444-(hifluommethylhenllimidazole The imine prepared in Example 1(c) was reacted with the isocyanide prepared in the previous step using the procedure of example 1(d) to prepare the title compound was prepared. ESMS =417 [M+H] Examle 69 I -(CvclopLQbvlmmgbl .4-dichloroDhenvl)-5-14--bldinyl)imidazol a) 3.4-ichlorohenX-tollthiomthlisocvide Following the procedure of example 1(ab) except using 3.4-dicblorobenzaldehyde as the aldehyde component the title compound was prepared.
b) I- CvlogMIvlmthvl)4-(3 44-dichloroahenvl-5-44bvrdinvlimidazol Following the procedure of example 1(d) substituting the imne prepared in Example 27(a) and the isocyanide prepared in the previous step the title compound was prepared. mp 14550.
Example 1' -(CgiolrolimthllA:(-trfluomet4l(enyl-S-4zpjioiyommidzol Following the procedure of Example 1(d) substituting the imine prepared in Example 27(a) and the isocyanide prepared Example 68(a) the title compound was prepared. mp 105.50.
Example 71 a) 2-Methylovridine4!caeoxlddhLde (gvlooaw= tvlim~ Reaction of 4-fonnyl-2-methylpyridine [prepared in Example 33(a)] and cyclopropylmethyl amine by the procedure of example 1(c) affords the title compound as a yellow oil.
b) 1 lg=Mdmethv44fu (4-n--2-oahd4vlmdzl Following the procedure of example 1(d) except using the compound of the 46 previous step as the imine afforded the title compound was as a white solid (62% from the 2-aminopyrimidine aldehyde). mp 141.0 141.50.
Examile 72 1 -f3-(4-MoholinynDyropl11 5-(4-yi dinvn)-4-(3 a) 3--bisrifl uoromethylbhenyj-talylthiomethylisocvanW Following the procedure of example I(ab) except using benzaldehyde as the aldehyde component the title compound was prepared.
b) I -r3(4-Morphoinvl)ropvfl-5-(4-ovridiivl)-4-(3.5-bistrifluoromethynblnyc)imidazole Following the procedure of example 1(d) substituting the imine prepared in Example 1(c) and the isocyanide prepared in the previous step the title compound was prepared. mp 136.5-137.50.
Examle 73 5-r4-(2-Aminpyriiidinyl)1-4-4-fluorophen l- 1 -(2-carboxy-2.2-dimethyiethyli.
a) 2-Aminovrimidine-4-csrboxaldehvde (ethyl 3-imino-2-2-dimethylprogionate)imine 2-Aminopyrimidine-4-carboxaidehyde and ethyl 3-amino-2,2-dimethylpropionate, were reacted by the procedure of example 1(c) to afford the title compound as a yellow oil.
b) 5-r1(2-Aming vri idivlll-4 CefluoEo beevr: 1 -(2-carboxyethyl-2iehlrblmdze Following the procedure of example I(d) except using the compound of the previous step as the imine afforded the title compound as a white solid (I11% from the amine).
c) 5-r4-(2-Aminolyrimidiny)1 -444-flurophena- I 42-carbo-2-2dimethylethyjimidazole lithium salt The compound of example 73(c) was hydrolysed by the procedure of example 37 to afford the title compound as the lithium salt; ES MS mie 356.
Examle 74 I-(I -Formyl-Q4-piprdinf-4-(4-fluooohenyfl-5-(4-pvriimidazole 1-(l-Benzylpiperidin-4-yl)-4-(4-fluorophenyl-5-(4-pyridyl)imidazole (100 mg;prepared in Example 40) was dissolved in 10% formic acidhnethanol under argon and palladium black (100 mg) mixed in 10% formic acid/methanol was added. The 47 reaction was stirred under argon at room temperature for sixteen hours. The reaction mixture was evaporated and the residue mixed in H20/etiyl acetate and the PH taken -to 10. The layers were separated and the aqueous phase extracted with ethyl acetate.
The combined organic layers were evaporated and the residue was flash chrornatographed (silica gellrnethylene chloride/methanol) to yield the title compound, an off-white solid. ES MS Wue 351 (MH+) Example 7 5-(2-Amnino-4-Dvrimidinvl).4-4-.fluoronhenyji). -(l-methyj-4-DipenidinvflimidaznIe a) 4-Amino- I-methylpipgridine 1-Methylpiperidin-4-one (4.22 g, 37 mrnol) and an ice cold soln of !N HCI in (37 mil, 37 mirol) were combined. Trituration followed by evaporation of the Et 2 O at 230 under a stream of argon afforded the hydrochloride. MeOH (114 mQL, anhydrous NH4OAc (28.7, 373 nimol) and 3A molecular sieves were added. Stirred min and then NaCNBH 3 (2.33 g, 37 nimol) was added, and the mixture was Stirred for 1 h. Acidified to pH 1 with concentrated HCI and washed with Et 2 o. The resulting mixture was made basic with 50% aq NaOH and extracted with EtOAc, dried (K 2 C0 3 and distilled (bp 55 -600, 15 mm) to afford 3.88 g Of the title compound.
2-Aminopyrirnidine-4-carboxaldehyde and the compound of the previous, step were reacted by the Procedure Of example I1(c) to afford the title compound as a yellow oil.
yfiia~ Following the procedure of example 1(d) except using the compound of the Previous step as the imine afforded the title compound after purification by chromatography on silica with 0 10% MCOH and 0 Et3N in CH2Cl 2 followed by fractional preciptation from MeOH with Et20; as a yellow solid (20% from the amine).
nip 235 2370 (dcc).
EamplIC 7 glrmdnvlMmidazole a) I-Amino-2.2-dimetbvlppymorpholine 2.2-dimethyl-3-N-morpholinyl propionaldehyde (Cheney,LL J. Amer. Chem.
Soc. 1951, 73, p685 686; 855 mig, 5.0 nimol) was dissolved in Et2O (2 mL) and 1 N HCI in Et2O (5rnL, 5mmol) was added. Stirred 5min and the Et2O was evaporated in a streamn of Ar. The solid was dissolved in anhydrous MeOH (15 mQL followed by anhydrous NH4OAc (3.85 g, 50 nimol), and 3A molecular sieves. Stirred 5 min and then NaCNBH 3 (0.3 14 g, 4.0 nimoL) was added. Stirred 45 min and concd HICl was added til the reaction mixture was pH 1. The MeOH was removed in vacuo and the residual mixture was dissolved in H 2 0 (15 mL) and extracted with Et 2 0 (25 mL). The aq phase was layered with another portion of Et 2 0 and made basic by addition of aq NaOH til pH 10. Extraction with Et 2 0 (3 x 40 mL), drying (K2C0 3 and concentration afforded the title compound b) 2 -Aminnovrimidine-4-.carboxaldehyde 13-(4-Momhoinvl)-2.2dimethvioropyllimine 2 -Aminopyrimidine-4-carboxaldehyde and the product of the previous step were reacted by the procedure of example 1(c) to afford the title compound as a yellow oil.
c) Xrimidinvlimidazle Following the procedure of example 1(d) except using the compound of the previous step as the imine afforded the title compound as a white solid (16% from the amine). mp 242 245o (dec.).
Example 77 4-( 4 -Fluorohenvl-5-4-nvridvl) -1 2 -acetoxvethvilimidarole 500 mg of 4 4 -Fluorophenyl)-5-(4-pyridyl)imidazole was dried at 50' C overnight in vacuo and added to a flask containing 20 ml of dried (sieve) dimethyl formamide :(hereinafter DMF) and treated with NaH (at 0 then stirred at room temperature, and dropwise with 2-acetoxy ethylbromide. After three days, the mixture was poured into ice water, extracted into methyoene chloride, the organic phase washed with water, S* dried over sodium sulfate and stripped in vacuo. Flashed the residue on silica using CH2Cl2-acetone (85:15) and eluting with increasing CH30H from 0 to 10%. Two 'S major product fractions were obtained, the pure cuts combined to give a slower eluting :fraction and a faster eluting isomer. These isomers were stripped and recrystallized from EtOAc-hexane to give the minor isomer (slower moving) and the fast, major isomer (the titled compound). NMR (250mHz, CDCl3) shows CH2CH2 as singlet at 8 4.1 ppm, very clean, H-ortho to F, triple t at 6.9 ppm. Cal'd C:66.60, H:4.86, N:12.92; Found C:67.10, 67.03 H:5.07, 4.94 N:13.08, 13.09. IR (nujol mull) shows 1740 cm-1 (sharp, ester).
METHODS
QOFTREATMENT
The compounds of Formula or a pharmaceutically acceptable salt thereof can be used in the manufacture of a medicament for the prophylactic or therapeutic treatment of any disease state in a human, or other mammal, which is exacerbated or caused by excessive or unregulated cytokine production by such mammal's cell, such as but not limited to monocytes and/or macrophages.
49 Compounds of Formula are capable of inhibiting proinflammatory cyzokines, such as IL-1, IL-6, IL-8 and TNF and are therefore of use in therapy. IL-1, IL-6, IL-8 and TNF affect a wide variety of cells and tissues and these cytokines, as well as other leukocyte-derived cytokines, are important and critical inflammatory mediators of a wide variety of disease states and conditions. The inhibition of these pro-inflammatory cytokines is of benefit in controlling, reducing and alleviating many of these disease states.
Accordingly, the present invention provides a method of treating a cytokinemediated disease which comprises administering an effective cytokine-interfering amount of a compound of Formula or a pharmaceutically acceptable salt thereof.
In particular, compounds of Formula or a pharmaceutically acceptable salt thereof are of use in the prophylaxis or therapy of any disease state in a human, or other mammal, which is exacerbated by or caused by excessive or unregulated IL-1, IL-8 or TNF production by such mammal's cell, such as, but not limited to, monocytes and/or 15 macrophages.
Accordingly, in another aspect, this invention relates to a method of inhibiting the production of IL- in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula or a pharmaceutically acceptable salt thereof.
20 There are many disease states in which excessive or unregulated IL-1 production is implicated in exacerbating and/or causing the disease. These include rheumatoid arthrits ostorthritis, endotoxemia and/or toxic shock syndrome, other Sacute or chronic inflammatory disease states such as the inflammatory reaction induced by endotoxin or inflammatory bowel disease, tuberculosis, atherosclerosis, muscle degeneration, multiple sclerosis, cachexia, bone resorption, psoriatic arthritis, Reiter's Ssyndrome, rheumatoid arthritis, gout, traumatic arthritis, rubella arthritis and acute synovitis. Recent evidence also links IL- activity to diabetes, pancreatic 8 cells and Alzheimes disease.
In a further aspect, this invention relates to a method of inhibiting the production of TNF in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula or a pharmaceutically acceptable salt thereof.
Excessive or unregulated TNF production has been implicated in mediating or exacerbating a number of diseases including rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty arthritis and other arthritic conditions, sepsis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, adult respiratory distres syndrome, cerebral malaria, chronic pulmonary inflammatory disease, silicosis, pulmonary sarcoisosis, bone resorption diseases, such as osteoporosis, reperfusion injury, graft vs. host reaction, allograft rejections, fever and myalgias due to infection, such as influenza, cachexia secondary to infection or malignancy, cachexia secondary to acquired immune deficiency syndrome (AIDS), AIDS, ARC (AIDS related complex), keloid formation, scar tissue formation, Crohn's disease, ulcerative colitis and pyresis.
Compounds of Formula are also useful in the treatment of viral infections, where such viruses are sensitive to upregulation by TNF or will elicit TNF production in vivo. The viruses contemplated for treatment herein are those that produce TNF as a result of infection, or those which are sensitive to inhibition, such as by decreased replication, directly or indirectly, by the TNF inhibiting-compounds of Formula Such viruses include, but are not limited to HIV-1, HIV-2 and HIV-3, Cytomegalovirus (CMV), Influenza, adenovirus and the Herpes group of viruses, such as but not limited to, Herpes Zoster and Herpes Simplex. Accordingly, in a further aspect, this invention relates to a method of treating a mammal afflicted with a human immunodeficiency virus (HIV) which comprises administering to such mammal an effective TNF 15 inhibiting amount of a compound of Formula or a pharmaceutically acceptable salt thereof.
Compounds of Formula may also be used in association with the veterinary treatment of mammals, other than in humans, in need of inhibition of TNF production.
TNF mediated diseases for treatment, therapeutically or prophylactically, in animals 20 include disease states such as those noted above, but in particular viral infections.
Examples of such viruses include, but are not limited to, lentivirus infections such as, equine infectious anaemia virus, caprine arthritis virus, visna virus, or maedi virus or retrovirus infections, such as but not limited to feline immunodeficiency virus (FIV), bovine immunodeficiency virus, or canine immunodeficiency virus or other retroviral infections.
The compounds of Formula may also be used topically in the treatment or prophylaxis of topical disease states mediated by or ex erbated by excessive cytokine production, such as by IL-1 or TNF respectively, such as inflamed joints, eczema, psoriasis and other inflammatory skin conditions such as sunburn; inflammatory eye conditions including conjunctivitis; pyresis, pain and other conditions associated with inflammation.
Compounds of Formula have also been shown to inhibit the production of IL-8 (Interleukin-8, NAP). Accordingly, in a further aspect, this invention relates to a method of inhibiting the production of IL-8 in a mammal in need thereof which comprises administering to said mammal an effective amount of a compound of Formula or a pharmaceutically acceptable salt thereof.
There are many disease states in which excessive or unregulated IL8 production is implicated in exacerbating and/or causing the disease. These diseases are characterized by massive neutrophil infiltration such as, psoriasis, inflammatory bowel 51 disease, asthma, cardiac and renal reperfusion injury, adult respiratory distress syndrome, thrombosis and glomerulonephritis. All of these diseases are associated with increased IL-8 production which is responsible for the chemotaxis of neutrophils into the inflammatory site. In contrast to other inflammatory cytokines
TNF,
and IL-6), IL-8 has the unique property of promoting neutrophil chemotaxis and activation. Therefore, the inhibition of IL-8 production would lead to a direct reduction in the neutrophil infiltration.
The compounds of Formula are administered in an amount sufficient to inhibit cytokine, in particular IL-1, IL-6, IL-8 or TNF, production such that it is regulated down to normal levels, or in some case to subnormal levels, so as to ameliorate or prevent the disease state. Abnormal levels ofIL-1, IL-6, IL-R or TNF, for instance in the context of the present invention, constitute: levels of free (not cell bound) IL-, IL-6, IL-8 or TNF greater than or equal to I picogram per ml; (ii) any cell associated IL-1, IL-6, IL-8 or TNF; or (iii) the presence of IL-, IL-6. IL-8 or TNF 15 mRNA above basal levels in cells or tissues in which IL-1, IL-6, IL-8 or TNF, respectively, is produced.
The discovery that the compounds of Formula are inhibitors of cytokines, specifically IL-1, IL-6, IL-8 and TNF is based upon the effects of the compounds of Formulas on the production of the IL-1, IL-8 and TNF in in vitro assays which are 20 described herein.
As used herein, the term "inhibiting the production of IL-1 (IL-6, IL-8 or TNF)" refers to: a) a decrease of excessive in vivo levels of the cytokine (IL-1, IL-6, IL-8 or TNF) in a human to normal or sub-normal levels by inhibition of the in vivo release of the cytokine by all cells, including but not limited to monocytes or macrophages; b) a down regulation, at the genomic level, of excessive in vivo levels of the cytokine (IL-1, IL-6, IL-8 or TNF) in a human to normal or sub-normal levels; c) a down regulation, by inhibition of the direct synthesis of the cytokine (IL-1, IL-6, IL-8 or TNF) as a postranslational event; or d) a down regulation, at the translational level, of excessive in vivo levels of the cytokine (IL-1, IL-6, IL-8 or TNF) in a human to normal or sub-normal levels.
As used herein, the term "TNF mediated disease or disease state" refers to any and all disease states in which TNF plays a role, either by production of TNF itself, or by TNF causing another monokine to be released, such as but not limited to IL-1, IL-6 or IL-8. A disease state in which, for instance, IL- I is a major component, and whose production or action, is exacerbated or secreted in response to TNF, would therefore be considered a disease stated mediated by TNF.
52 As used herein, the term "cytokine" refers to any secreted polypeptide that atfects the functions of cells and is a molecule which modulates interactions between cells in the immune, inflammatory or hematopoietic response. A cytokine includes, but is not limited to, monokines and lymphokines, regardless of which cells produce them.
For instance, a monokine is generally referred to as being produced and secreted by a mononuclear cell, such as a macrophage and/or monocyte. Many other cells however also produce monokines, such as natural killer cells, fibroblasts, basophils, neutrophils, endothelial cells, brain astrocytes, bone marrow stromal cells, epideral keratinocytes and B-lymphocytes. Lymphokines are generally referred to as being produced by lymphocyte cells. Examples of cytokines include, but are not limited to, Interleukin- Interleukin-6 Interleukin-8 Tumor Necrosis Factor-alpha (TNFa) and Tumor Necrosis Factor beta (TNF-B).
As used herein, the term "cytokine interfering" or "cytokine suppressive amount" refers to an effective amount of a compound of Formula which will cause a 15 decrease in the in vivo levels of the cytokine to normal or sub-normal levels, when given to a patient for the prophylaxis or treatment of a disease state which is exacerbated by, or caused by, excessive or unregulated cytokine production.
As used herein, the cytokine referred to in the phrase "inhibition of a cytokine, for use in the treatment of a HIV-infected human" is a cytokine which is implicated in the initiation and/or maintenance of T cell activation and/or activated T cellmediated HIV gene expression and/or replication and/or any cytokine-mediated dis ese associated problem such as cachexia or muscle degeneration.
As TNF-B (also known as lymphotoxin) has close structural homology with TNF-a (also known as cachectin) and since each induces similar biologic responses and binds to the same cellular receptor, both TNF-a and TNF-8 are inhibited by the compounds of the present invention and thus are herein referred to collectively as "TNF" unless specifically delineated otherwise.
In order to use a compound of Formula or a pharmaceutically acceptable salt thereof in therapy, it will normally be Formulated into a pharmaceutical composition in accordance with standard pharmaceutical practice. This invention, therefore, also relates to a pharmaceutical composition comprising an effective, non-toxic amount of a compound of Formula and a pharmaceutically acceptable carrier or diluent.
Compounds of Formula pharmaceutically acceptable salts thereof and pharmaceutical compositions incorporating such may conveniently be administered by any of the routes conventionally used for drug administration, for instance, orally, topically, parenterally or by inhalation. The compounds of Formula may be administered in conventional dosage forms prepared by combining a compound of Formula with standard pharmaceutical carriers according to conventional procedures. The compounds of Formula may also be administered in conventional 53..
dosages in combination with a known, second therapeutically active compound. These procedures may involve mixing, granulating and compressing or dissolving the ingredients as appropriate to the desired preparation. It will be appreciated that the form and character of the pharmaceutically acceptable character or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables. The carrer(s) must be "acceptable" in the sense of being compatible with the other ingredients of the Formulation and not deleterious to the recipient thereof.
The pharmaceutical carrer employed may be, for example, either a solid or liquid. Exemplary of solid carriers are lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Exemplary of liquid carriers are syrup, peanut oil, olive oil, water and the like. Similarly, the carrer or diluent may include time delay material well known to the art, such as glyceryl mono- 0* *stearate or glyceryl disteanate alone or with a wax.
15 A wide variety of pharmaceutical forms can be employed. Thus, if a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet formi or in the form of a troche or lozenge. The amount of solid carrier will vary widely but preferably Will be from about 25mg. to about I1g. When a liquid carrier is used, the preparation will be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable liquid such as an ampule or nonaqueous liquid suspension.
S Compounds of Formula may be administered topically, that is by nonsystemic administration, Ibis includes the application of a compound of Formula (I) externally to the epidermis or the buccal cavity and the instillation of such a compound into the ear, eye and nose, such that the compound does not significantly enter the blood stream. In contrast, systemic administration refers to oral, intravenous, 11 :5 intraperitoneal and intramuscular administration.
Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site of inflammation such as liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose. The active ingredient may comprise for topical administration, from 0.001 to 10% w/w, for instance from 1% to 2% by weight of the Formulation.
It may however comprise as much as 10% w/w but preferably will comprise less than wlw, more preferably from 0. 1% to I% w/w of the Formulation.
Lotions according to the present invention include those suitable for application to the skin or eye. An eye lotion may comprise a sterile aqueous solution optionally containing a bactericide and may be prepared by methods similar to those for the preparation of drops. Lotions or liniments for application to the skin may also include 54.
an agent to hasten drying and to cool the skin, such as an alcohol or acetone, and/or a moisturizer such as glycerol or an oil such as castor oil or arachis oil.
Creams, ointments or pastes according to the present invention are semi-solid Formulations of the active ingredient for external application. They may be made by mixing the active ingredient in finely-divided or powdered form, alone or in solution or suspension in an aqueous or non-aqueous fluid, with the aid of suitable machinery, with a greasy or non-greasy base. The base may comprise hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, a metallic soap; a mucilage; an oil of natural origin such as almond, corn, arachis, castor or olive oil; wool fat or its derivatives or a fatty acid such as steric or oleic acid together with an alcohol such as propylene glycol or a macrogeL The Formulation may incorporate any suitable surface active agent such as an anionic, cationic or non-ionic surfactant such as a sorbitan ester or a polyoxyethylene derivative thereof. Suspending agents such as natural gums, cellulose derivatives or inorganic materials such as silicaceous silicas, and other ingredients such 15 as lanolin, may also be included.
-Drops according to the present invention may comprise sterile aqueous or oily solutions or suspensions and may be prepared by dissolving the active ingredient in a suitable aqueous solution of a bactericidal and/or fungicidal agent and/or any other suitable preservative, and preferably including a surface active agent The resulting 20 solution may then be clarified by filtration, transferred to a suitable container which is then sealed and sterilized by autoclaving or maintaining at 98-100 C. for half an hour.
Alternatively, the solution may be sterilized by filtration and transferred to the container by an aseptic technique. Examples of bactericidal and fungicidal agents suitable for inclusion in the drops are phenylmercuric nitrate or acetate (0.002%), benzalkonium chloride and chlorhexidine acetate Suitable solvents for the preparation of an oily solution include glycerol diiuted alcohol and propylene glycol.
Compounds of formula may be administered parenterally, that is by intravenous, intramuscular, subcutaneous intranasal, intrarectal, intravaginal or intraperitoneal administration. The subcutaneous and intramuscular forms of parenteral administration are generally preferred. Appropriate dosage forms for such administration may be prepared by conventional techniques. Compounds of Formula may also be administered by inhalation, that is by intranasal and oral inhalation administration. Appropriate dosage forms for such administration, such as an aerosol Formulation or a metered dose inhaler, may be prepared by conventional techniques.
For all methods of use disclosed herein for the compounds of Formula the daily oral dosage regimen will preferably be from about 0.1 to about 80 mg/kg of total body weight, preferably from about 0.2 to 30 mg/kg, more preferably from about mg to 15mg. The daily parenteral dosage regimen about 0.1 to about 80 mg/kg of total body weight, preferably from about 0.2 to about 30 mg/kg, and more preferably from agout 0.5 mg to 15mg/kg. The daily topical dosage regimen will preferably be from 0.1 mg to 150 mg, administered one to four, preferably two or three times daily. The daily inhalation dosage regimen will preferably be from about 0.01 mg/kg to about I mg/kg per day. It will also be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a compound of Formula or a pharmnnaceutically acceptable salt thereof will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the particular patient being treated, and that such optimums can be determined by conventional techniques. It will also be appreciated by one of skill in the art that the optimal course of treatment, the number of doses of a compound of Formula or a pharmaceutically acceptable salt thereof given per day for a defined number of days, can be ascertained by those skilled in the art using conventional course of treatment determination tests.
15 The invention will now be described by reference to the following biological examples which are merely illustrative and are not to be construed as a limitation of the scope of the present invention.
BIOLQCATL EXAMPLIp
M\
20 The cytokine-inhibiting effects of compounds of the present invention were "determined by the following in vitro assays: Interleukln. 1 (IL-) Human peripheral blood monocytes were isolated and purified from either fresh "blood preparations from volunteer donors, or from blood bank buffy coats, according to the procedure of Colotta et a, J Immunol, 132, 936 (1984). These monocytes (Ixl0 6 were plated in 24-well plates at a concentration of 1-2 million/ml per well. The cells were allowed to adhere for 2 hours, after which time non-adherent cells were removed by gentle washing. Test compounds were then. added to the cells for lh before the addition of lipopolysaccharide (50 ng/ml), and the cultures were incubated at 37 0 C for an additional 24h. At the end of this period, culture super-natants were removed and clarified of cells and all debris. Culture supenatants were then immediately assayed for IL- 1 biological activity, either by the method of Simon et al., J. ImmunoL Methods, 84, 85, (1985) (based on ability of IL- 1 to stimulate a Interleukin 2 producing cell line (EL-4) to secrete IL-2, in concert with A23187 ionophore) or the method of Lee et aL, J. ImmunoTherapy, 6 1-12 (1990) (EUSA assay). The compounds of Frrmula as evidenced by Examples 1 to 24 were shown to be inhibitors of in vitro IL-I produced by human monocytes.
56 Tumour Necrosis Factor (TNF): Human peripheral blood monocytes were isolated and purified from either blood bank buffy coats or plateletpheresis residues, according to the procedure of Colotta, R. et al., J Immunol, 132(2), 936 (1984). The monocytes were plated at a density of Ix106 cells/ml medium/well in 24-well multi-dishes. The cells were allowed to adhere for 1 hour after which time the supernatant was aspirated and fresh medium (Iml, RPMI-1640, Whitaker Biomedical Products, Whitaker, CA) containing 1% fetal calf serum plus penicillin and streptomycin (10 units/ml) added. The cells were incubated for 45 minutes in the presence or absence of a test compound at InM- 10mM dose ranges (compounds were solubilized in dimethyl sulfoxide/ethanol, such that the final solvent concentration in the culture medium was 0.5% dimethyl ethanol). Bacterial lipopoly-saccharide coli 055:B5 [LPS] from Sigma Chemicals Co.) was then added (100 ng/ml in 10 ml phosphate buffered saline) and cultures incubated for 16-18 hours at 37°C in a 5% CO 2 incubator. At the end of 15 the incubation period, culture supernatants were removed from the cells, centrifuged at 3000 rpm to remove cell debris. The supernatant was then assayed for TNF activity using either a radio-immuno or an ELISA assay, as described in WO 92/10190 and by Becker et al., J Immunol, 1991, 147, 4307. The compounds of Formula as evidenced by Examples 1 to 24 were shown to be inhibitors of in vitro TNF produced 20 by human monocytes.
IL-
1 and TNF inhibitory activity does not seem to correlate with the property of the compounds of Formula in mediating arachidonic acid metabolism inhibition.
Further the ability to inhibit production of prostaglandin and/or leukotriene synthesis, by nonsteroidal anti-inflammatory drugs with potent cyclooxygenase and/or lipoxygenase inhibitory activity does not mean that the compound will necessarily also inhibit TNF or IL-I production, at non-toxic doses.
Interleukin -8 (IL-8): Primary human umbilical cord endothelial cells (HUVEC) (Cell Systems, Kirland, Wa) are maintained in culture medium supplemented with 15% fetal bovine serum and 1% CS-HBGF consisting of aFGF and heparin. The cells are then diluted before being plated (250pl) into gelating coated 96-well plates. Prior to use, culture medium are replaced with fresh medium (200dl). Buffer or test compound (25gl, at concentrations between I and 10M) is then added to each well in quadruplicate wells and the plates incubated for 6h in a humidified incubator at 37 0 C in an atmosphere of 5% C02. At the end of the incubation period, supernatant is removed and assayed for IL-8 concentration using an IL-8 ELISA kit obtained from R&D Systems (Minneapolis, MN). All data is presented as mean value (ng/ml) of 57 multiple samples based on the standard curve. IC50's where appropriate are generated by non-linear regression analysis.
Cytokine Specific Binding Protein Assay A radiocompetitive binding assay was developed to provide a highly reproducible primary screen for structure-activity studies. This assay provides many advantages over the conventional bioassays which utilize freshly isolated human monocytes as a source of cytokines and ELISA assays to quantify them. Besides being a much more facile assay, the binding assay has been extensively validated to highly correlate with the results of the bioassay. A specific and reproducible CSAIDTh cytokine inhibitor binding assay was developed using soluble cystosolic fraction from THP.I cells and a radiolabeled compound. For instance, a suitable radiolabeled compound of this CSAID m cytokine inhibitor class is 4-(Fluorophenyl)-2-(4hydroxyphenyl-35-t2)-5-(4-pyridyl)imidazole. In brief, the THP. 1 cytosol was 15 routinely prepared from cell lysate obtained by nitrogen cavitation followed by a 10 K x g low speed and a 100 K x g high speed centrifugation, the supernatant of which was designated as the cytosolic fraction. THP.1 cytosol was incubated with appropriately diluted radioligand at room temperature for a pre-determined time to allow the binding to achieve equilibrium. The sample was added to a G-10 column and eluted with 20 mm TRN, 50mMb mercaptoethanol, NaN 3 The fraction encompassing the void volume was collected and the radioactivity was assessed by liquid scintillation counting. This was determined to reflect bound radioligand since the radioactive signal was abrogated by the presence of excess cold ligand in the incubation mixture or when there was no cytosolic fraction present. Compounds of Formula at various doses were added to the binding assay to achieve inhibition of binding of the radiolabel.
IC50s as well as Ki values were determined by regression analysis and scatchard plot analysis respectively. There is generally excellent correlation between the IC50 of compounds tested in both the binding assay and the bioassay and can be used interchangeably in many cases.
Patent Application USSN 08/123175 Lee et aL, filed September 1993 whose disclosure is incorporated by reference herein in its entirey describes the above noted method for screening drugs to identify compounds which interact with and bind to the cytokine specific binding protein (hereinafter CSBP). However, for purposes herein the binding protein may be in isolated form in solution, or in immobilized form, or may be genetically engineered to be expressed on the surface of recombinant host cells such as in phage display system or as fusion proteins. Alternatively, whole cells or cytosolic fractions comprising the CSBP may be employed in the creening protocol. Regardless of the form of the binding protein, a plurality of compounds are contacted with the binding protein under conditions sufficient to form a compound/ binding protein 58 complex and compound capable of forming, enhancing or interfering with said complexes are detected.
More specifically, the Binding Assay is performed as follows:
MAEIAS
Incubation buffer: 20 mM Tris, I MuM M9C0 2 20 mM Hepes, 0.02% NaN3, store at Elution buffer 20 mM Tmi, 50 mM 2-mercaptoethanol, NaN 3 Store at 4C.
Sephadex: add 100 g Sephadex G-10 (Phannacia, Uppsala, Sweden) to 400 ruL dd H 2 0 and allow to swell at room temperature for 2 hours. Decant fines and wash 3 times. Add NaNJ and qs with dd H 2 0 to 500 ruLs and store at 4*C.
Assemble Columns: Straw column, filter fit a nd tip (Kontes, SP.420160-000, 420162 002). Lowsorb tubes (Nunc) used in binding reaction. THP.1I cytosol spun at 15000 rpm for 5 nin to clarify. THP. 1 cytosol prepared by hypnotic treatment of cells and lysis by decompression in nitrogen. Nuclei and membrane fragments removed by :0.15 differential centrifugation (10,000 g for I hour and 100,000 g for 1 hour).
Comundi: Non-radioactive Compound I with corresponding EtOH control (dilutions made in incubation buffer) and 3 H-Compound I (dilutions in incubation buffer) A. Column Preparation I 1. Begin 30 muin before anticipated elution of reaction mixture.
2. Add 3mL of G-10 slurry to column for bed vol of 1.5 mL.
Rinse with 7 ruL elution buffer (fill to top of column) 4. Cut columns down to size.
B. Sample Incubation 1. 15 ruin incubation at4C.
Binding reaction mixture; 100 p.L cytos&A 10 uL cold Compound I or EtOH control 104Ii 3 H-Compouncl I (molar concentration depends on nature of study).
3. "Free" control 1004~± incubation buffer in lieu of cytosol preparation.
C. Sample Elution 1. Elute at 4 0
C.
2. Add total reaction volume to G- 10 column.
3. Add 400 gJ. elution buffer to column and discard eluate.
4. Add 500 p±L elution buffer to column, collecting eluted volume in ml scintillation vial.
Add 15 ruL Ready Safe scintillation fluid.
6. Vortex and count in liquid scintillation counter for 5 minutes.
59 Include a "total input counts control" (10 tL of labeled ligand).
D. Data Analysis I. Plot DPMS as ouptut in graphic form and analyze by regression analysis and "Lundon ligand binding" software for the determination of IC 50 and Kd/Ki respectively.
2. Rank order the IC50s of the tested compounds in the bioassay and compare to that generated by the binding assay and establish a correlation curve.
The binding assay was further validated by the following criteria: THP.1 cytosol demonstrated saturable and specific binding of the radiolabeled compound.
Preparation of 4-(Fluorophenyl)-2-(4-hydroxyphenyl-3,5-t2)-5-(4-pyridyl)imidazole, (Compound
I).
o 15 A 2.9 mg (0.0059 rmmol) portion of 2 3 ,5-Dibromo-4-hydroxyphenyl)-4-( 4 fluorophenyl)-5-(4-pyridyl)imidazole, Compound was dissolved in 0.95 mL of dry DMF and 0.05 mL of triethylamine in a 2.4 mL round bottom flask equipped with a small magnetic stirring bar. A 1.7 mg portion of 5% Pd/C (Engelhard lot 28845) was added, and the flask was attached to the stainless steel tritium manifold. The mixture 20 was degassed through four freeze-pump-thaw cycles, then tritium gas (5.3 Ci, 0.091 mmol) was introduced. The reaction mixture was allowed to warm to room temperature and was stirred vigorously for 20h. The mixture was frozen in liquid nitrogen, the remaining tritium gas (2.4 Ci) was removed, and the flask was removed from the manifold. The reaction mixture was transferred, using 3 x 1 mL of methanol as rinsings, into a 10 mL round bottom flask, and the solvents were removed by static vacuum transfer. A 1.5 mL portion of methanol was added to the residue, then removed by static vacuum transfer. The latter process was repeated. Finally, the residue was suspended in mL of ethanol and filtered through a syringe-tip Millipore filter (0.45 micron), along with 3 x ca. 1 mL ethanol rinsings. The total filtrate volume was determined to be 3.9 mL, and the total radioactivity, 94.2 mCi. Solution was determined to be 3.9 mL, and the total radioactivity, 94.2 mCi HPLC analysis of filtrate (Partisil 5 ODS-3, 4.6 mm I.D. x cm, 1 mUL/min of 70:30:01 water/acetonitrile/trifluoroacetic acid, Radiomatic Flo-One Beta radio detector with 3 mUmin of Ecoscint-H cocktail through a 0.75 mL cell) showed the presence of Compound I (R t 60 min. ca. 37% of total radioactivity), and a discrete intermediate presumed to be the monobromo derivative Compound la (Rt 11.8 min. ca. The filtrate solution was evaporated to near dryness with a stream of nitrogen, and the residue was dissolved in about 1.2 mL of the HPLC mobile phase. The solution was separated by HPLC as shown below, and rue peaks corresponding to Compounds
I
and la and SB collected separately.
HPLCMtho Column Altex Ulzrasphere mmIJ.D. x25 cm Mobile Phase 70:30:0.1 water/ace conitrildzrjfluoroaceti acid Flow Rate 5 m~lmin UV detection 2 l0ni Injection Volumes 0.05 0.4 mn: Retention Times 7.8 min Compound
I
24 min Compound la *.:The pooled Compound I fractions totaled 32 niL in volume and the radioactive concentration was 1.52 rnCi/inL (total 48.6 mn Ci). The pooled SB Compound la Oujn fractions (totaling 10. 1 mCi) were evaporated to dryness and the residue was transferred quantitatively into a glass vial using 3.8 niL of absolute ethanol for further analysis.
An 8 niL (12.2 mCi) portion of Compound I was evaporated to dryness in vacuo at <35"C, then redissolved in 0.5 niL of mobile phase. The whole volume was injected into the HPLC system described above, and the appropriate peak was collected. Evaporation of the collected eluate in vacuo at <35C and trnser of the yellow residue into a vial with absolute ethanol provided a solution (3.8 ml, 2.44 a iCi/mL) of Compound 1. The. portion of this solution used for NMR analyses was first evaporated to dryness using stream of nitrogen then taken up in CD 3
OD.
Analysis of 4 4 -Fuorophenyl).2-(4-hydroxyphenyl-3$>.52-5(4 pyridyl)iinidazole, Compound
L
RadiochemicAl Purity by HPLC Column Ultraspbere Octyl, 5mm, 4.6 mmn I.D. x 25 cm, Beckan Mobile Phase 350 :l50:O.5(vlvl/v) watacttied/dfuoroactic acid Flow Rate 1 .0 mLlmin Mass detection UV at 210 nm Radioactivity detection Raznona-D radioactivity flow detector Scintillator Tmu-Count CITru-Lab Supply Co.) Flow rate 5.0 mi~min Cell volume 0.75 mL Retention time 7.7 min Egul 2U2 61 Radioactive Concentration by Scintillation Countiny M2CIQ9 Scintillator Instrtiment Efficiency Resu Ready Safe (Beckman Instruments, Inc.) TM Analytic model 6881 Automated DPM calculation from quench curve 2.44- mCi/niL S~afc Activitv bV Mtqs Rrtmmmr,, Mcthod Resti CI-M,
NH
3 reagent gas 20.0 Cilminol 3Hi Distribution: Unlabeled 44% Single Label 43% Double Label 13%
S
S
*5 5 9 *5*S 5 *5 5 59 *9 3
HMR
9 Instrument Experiment Peak Referencing Solvent Brnker AM 400 Proton decoupled 3 H NMR Proton non-decoupled 3 H N1~1R Proton Don-decoupled 3 H NM[R Solvent Peak of methanol cl 3.3 Methanol-cl 4 Tritium is incorporated exclusively on the carbon atoms ortho to aromatic hydroxyl group Analvitical Rnm A Xvt~2 fL NNo-t, Radiochemical Purity determined by HPLC Radioactivity concentration determined by scintillation counting 98.7% 2.44 mCi/mL Specific activity determined by mass spewtrometzy 20.0 Ci/mmol 3 HNUR agrees with the proposed structure Representative compounds of Formula Examples 1 to 77, but for the compound of example 2- not tsted and compound of Example 72, have all demonstratedI a positive inhibitory activity in this binding away.
The above description fully discloses the invention including preferred embodiments thereof. Modifications and improvements of the embodiments specifically disclosed herein are within the scope of the following claims. Without further elaboration, it is believed that one skilled in the are can. using the preceding description, utilize the present invention to its fullest extent. Therefore the Examples herein are to be construed as merely illustrative and not a limitation of the scope of the present invention in any way. The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows.
P 'OPERMIC'73354-94 DIV- 12,6198
U
62 Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be urKierstood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.
a. a a
S.
a. a a.
*a

Claims (8)

1. A compound of the formula IHCHO R,4 IHCHO wherein R 4 is a phenyl substituted one more times independently by fluorine, SRS, SOR 5 or halo-substituted C 1 -4 alkyl; R 5 is hydrogen, C 1 -4 alkyl, C2 4 alkenyl, C2 4 alkynyl or NRyR 1 7 excluding the moieties SR 5 being SNR 7 R 1 7 and SOR 5 being SOH; R 7 and R 17 is each independently selected from hydrogen or C-4 alkyl or R 7 and RI7 together with the nitrogen to which they are attached form a heterocyclic ring of 5 to 7 members which ring optionally contains an additional heteroatom selected from oxygen, sulfur or NRIs; R 1 is hydrogen or C 1 -4 alkyl; R 12 is hydrogen or R16; R 1 5 is Rio or C(Z)-CI-4 alkyl; R 16 is CI- 4 alkyl, halo-substituted-Cl4 alkyl, or C 3 7 cycloalkyl; and Z is oxygen or sulfur.
2. The compound according to claim 1 wherein the R 4 phenyl is substituted by fluorine.
3. The compound according to claim 2 wherein the phenyl is substituted in the 4- position.
4. The compound according to Claim 1 wherein the R 4 phenyl is substituted by SR 5 or S(O)R
5 P:\OPER\PDB\2209391 .239 27/8/99 64 The compound according to claim 4 wherein R 5 is a C 1 2 alkyl.
6. The compound according to claim 5 wherein the phenyl is substituted in the 4- position.
7. The compound according to claim 1 wherein the R 4 phenyl is substituted by CF 3
8. The compound according to claim 7 wherein the phenyl is substituted in the 4- position. S S S *5 5, S* SO S *5 *S*5 S 5555 S'~S 4S5* S S .5 5.5. S S *S C *S S C S@ f4 *505 S. S $5 DATED this 27th day of August 1999 SmithKline Beecham Corporation. By its Patent Attorneys DAVIES COLLISON CAVE
AU44782/99A 1993-07-16 1999-08-27 Tri-substituted imidazoles having multiple therapeutic properties Abandoned AU4478299A (en)

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AU71850/98A AU705568C (en) 1993-07-16 1998-06-02 Tri-substituted imidazoles having multiple therapeutic properties
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