WO2004031401A2 - Novel tyrosine kinases inhibitors - Google Patents

Novel tyrosine kinases inhibitors Download PDF

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WO2004031401A2
WO2004031401A2 PCT/US2003/030931 US0330931W WO2004031401A2 WO 2004031401 A2 WO2004031401 A2 WO 2004031401A2 US 0330931 W US0330931 W US 0330931W WO 2004031401 A2 WO2004031401 A2 WO 2004031401A2
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Prior art keywords
methyl
phenyl
hydroxy
pyridin
ethylamino
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PCT/US2003/030931
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French (fr)
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WO2004031401A3 (en
Inventor
Mark D. Wittman
Neelakantan Balasubramanian
Upender Velaparthi
Kurt Zimmermann
Mark G. Saulnier
Peiying Liu
Xiaopeng Sang
David B. Frennesson
Karen M. Stoffan
James G. Tarrant
Anne Marinier
Stephan Roy
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Bristol-Myers Squibb Company
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Priority to AU2003282891A priority Critical patent/AU2003282891A1/en
Priority to EP03774510A priority patent/EP1545543A4/en
Publication of WO2004031401A2 publication Critical patent/WO2004031401A2/en
Publication of WO2004031401A3 publication Critical patent/WO2004031401A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
    • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
    • C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings

Definitions

  • the present invention relates generally to the field of tyrosine kinase enzyme inhibition using novel small molecules.
  • Tyrosine Kinases are a class of enzymes, which catalyze the transfer of the terminal phosphate of adenosine triphosphate to the phenolic hydroxyl group of a tyrosine residue present in the target protein.
  • Tyrosine kinases play a critical role in signal transduction for several cellular functions including cell proliferation, carcinogenesis, apoptosis, and cell differentiation (Plowman, G. D.; Ullrich, A.; Shawver, L. K.: Receptor Tyrosine Kinases As Targets For Drug Intervention. DNSP (1994) 7: 334-339). Therefore inhibitors of these enzymes would be useful for the treatment or prevention of proliferative diseases which are dependent on these enzymes.
  • Tyrosine kinases that have been implicated in these processes include Abl, CDK's, EGF, EMT, FGF, FAK, Flk-1/KDR, HER-2, IGF-1R, IR, LCK, MET, PDGF, Src, and VEGF (Traxler, P.M. Protein Tyrosine Kinase Inhibitors in Cancer Treatment. Exp. Opin. Ther. Patents (1997) 7: 571-588; incorporated herein by reference). Hence, there is an ongoing need to investigate novel compounds that can be used to regulate or inhibit tyrosine kinase enzymes.
  • the present invention relates to compounds which inhibit tyrosine kinase enzymes, compositions which contain tyrosine kinase inhibiting compounds and methods of using inhibitors of tyrosine kinase enzymes to treat diseases which are characterized by an overexpression or upregulation of tyrosine kinase activity such as cancer, diabetes, restenosis, arteriosclerosis, psoriasis, angiogenic diseases and immunologic disorders (Powis, G.; Workman, P. Signaling targets For The Development of Cancer Drugs. Anti-Cancer Drug Design (1994), 9: 263-211; Merenmies, J.; Parada, L. E; Henkemeyer, M.
  • X is selected from the group consisting of N, C, C ⁇ -C 3 alkyl, C ⁇ -C alkyl substituted with one or more R 7 , and a direct bond;
  • Y is selected from the group consisting of O and S ;
  • W is selected from the group consisting of N, C, O, and S; provided that if W is O or S, R 9 is absent;
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R , R 9 are each independently selected from the group consisting of H, C 1-6 alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, -OR 60 , -NO 2 , -OH, -SR 60 , -NR 60 R 61 , -CN, -CO 2 R 60 , -CONR 60 R 61 , OCONR 60 R 61 , -NR 62 CONR 60 R 61 , -NR 60 SO 2 R 61 , -SO 2 NR 60 R 61 , -SO 2 R 63 , - C(NR 62 )NR 60 R 61 , aryl, heteroaryl, -(CH 2
  • Z is selected from the group consisting of - C 4 alkyl, alkenyl, and alkynyl chain; Z having one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR 60 SO 2 R 61 groups; Z optionally incorporating one or more groups selected from the group consisting of CO, CNOH, CNOR 60 , CNNR 60 , CNNCOR 60 and CNNSO 2 R 60 ; and
  • R 60 , R 61 , R 62 , and R 63 are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy, alkoxy, aryl, heteroaryl, heteroarylalkyl, and alkyl-R 25 ;
  • R 25 is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, - NR 30 COOR 31 , -NR 30 C(O)R 31 , -NR 30 SO 2 R 31 , -C(O)NR 30 R 31 , heteroaryl or heterocycloalkyl; and R 30 and R 31 are, independently, hydrogen, alkyl, or cycloalkyl.
  • R 1 , R 7 , R 8 and R 9 are H; R 2 and R 4 are H or F; Y is O; X is selected from the group consisting of N and CH; W is N; R 5 is selected from the group consisting of H, methyl, ethyl, isopropyl, secondary butyl, cyclopropyl, F, and CF 3 ; R 6 is selected from the group consisting of H, 2-aminomethylpyridine,
  • R 3 is selected from the group consisting of OR 60 , C(NH)NHR 60 , C(O)NHR 60 imidazole, imidazoline, tetrahydropyrimidine, piperazine, morpholine, homomorpholine, piperidine, pyrrolidine, homopiperazine and amino; wherein R 60 is selected from the group consisting of H, alkyl, cycloalkyl, heterocycloalkyl, and alkyl-R 25 .
  • R 3 is -OR 60 and R 60 is alkyl, or -alkyl-R 25 , such as methyl, - (CH 2 ) complicatCH 2 OH, or -(CH 2 ) n CH 2 N(CH 2 CH 2 ) 2 O, and n is 0, 1, or 2.
  • R 25 is morpholine,
  • R 33 , R 34 , R 35 , R 36 , and R 37 are hydrogen, alkyl, or substituted alkyl.
  • R 3 is piperazine, homopiperazine, 3-methylpiperazine, or 3,5-dimethylpiperazine being optionally substituted at the 4-N position with a compound selected from the group consisting of alkyl, cycloalkyl, cycloalkylalkyl, alkyl-R 25 , -C(O)-R 15 , or -CO 2 R 15 wherein R 15 is hydrogen, alkyl, aryl, alkyl-R 25 , amino or aryl.
  • Preferred piperazines are substituted with methyl, ethyl, CH 2 -cyclopropyl, hydroxyethyl, 2-dimethylaminoethyl, 2- diethylaminoethyl, 2-aminoethyl, 2-methylaminoethyl, 2-ethylaminoethyl, methoxyethyl, ethoxyethyl, thiomethoxyethyl, morpholine, and morpholinylethyl.
  • R may be an amino group selected from the group consisting of hydroxyalkylamino, aminoalkylamino, dialkylaminoalkylamino, and heterocycloalkylalkylamino.
  • Preferred amino groups include NHCH 2 CH 2 OH, NMeCH 2 CH 2 OH, NEtCH 2 CH 2 OH, NHCH 2 CH 2 NH 2 , NMeCH 2 CH 2 NH 2 , NEtCH 2 CH 2 NH 2 , NHCH 2 CH 2 NMe 2 , NMeCH 2 CH 2 NMe 2 , NEtCH 2 CH 2 NMe 2 , NHCH 2 CH 2 NEt 2 , NMeCH 2 CH 2 NEt 2 , NEtCH 2 CH 2 NEt 2 , NHCH 2 CH 2 N(CH 2 CH 2 ) 2 O, NMeCH 2 CH 2 N(CH 2 CH 2 ) 2 O, and NEtCH 2 CH 2 N(CH 2 CH 2 ) 2 O.
  • R 3 is piperidine, optionally substituted with hydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, 1,3 dioxolane, 1,3 dioxane, -NHC(O)R 15 , -NHCO 2 R 15 , wherein R 15 is hydrogen, alkyl, aryl or alkyl-R 25 or morpholine, thiomorpholine, sulfoxymorpholine, sulfonylmorpholine, or homomorpholine, or a substituted morpholine, thiomorpholine, sulfoxymorpholine, sulfonylmorpholine, or homomorpholine.
  • the morpholine, thiomorpholine, sulfoxymorpholine, sulfonyl morpholine, or homomorpholine is optionally substituted with hydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, alkyl-R 25 , -NHC(O)R 15 , -NHCO 2 R 15 , wherein R 15 is hydrogen, alkyl, aryl or alkyl-R 25 .
  • R 3 is an optionally substituted pyrrolidine that may be selected from the group consisting of 3-hydroxyl pyrrolidine, 3- alkoxy pyrrolidine, and 3-alkylamino pyrrolidine, 3-dialkylamino pyrrolidine.
  • R 6 is selected from the group consisting of H, 2-aminomethylpyridine, NHCH 2 CH(OH)aryl, and
  • aryl groups include substituted phenyl wherein the phenyl is substituted with at least one Br, CI, F, -CN, methoxy, or-NHSO 2 CH 3 .
  • R is
  • R 17 is H, Br, CI, or F and R 40 is H or alkyl.
  • R 12 is hydrogen, methyl, hydroxymethyl, methoxymethyl, CH 2 F, CH 2 CN, CO 2 H, or -CONR 30 R 31 ;
  • R 13 is H or methyl;
  • R 17 is Br, F, or CI;
  • R 18 is methoxy or fluoro; and
  • R 19 is H; or R 18 and R 19 together form 4-O, 5 dihydrofuranyl.
  • compounds have the formula: wherein R 51 is hydrogen, alkyl, aryl, or -alkyl-R 25 and the remaining substituents are as defined above.
  • R 51 is hydrogen, methyl, ethyl, or (CH 2 ) n CH 2 -R 25 wherein R 25 is OH, OMe, F, CN, CF 3 , SOCH 3 or SO 2 CH 3 , wherein n is 0 or 1.
  • R 51 is methoxyethyl and R 6 is -NHCH 2 CHOH-aryl.
  • R 51 is cyanoethyl, hydroxyethyl, CH 2 CH 2 SOCH 3 , CH 2 CH 2 CH 2 F, CH 2 CH 2 CH 2 CN, or CH 2 CH 2 CF 3;
  • R 16 and R 19 are H;
  • R 17 is Br, or CI;
  • R 18 is hydrogen or methoxy.
  • R' 5 is hydrogen, alkyl, aryl or alkyl-R 25 ; each R 16 is independently hydrogen or methyl and the remaining substituents are as defined above.
  • R 15 is hydrogen or methyl;
  • R 17 is bromo, chloro or fluoro;
  • R 18 is hydrogen or methoxy; and
  • R 19 is hydrogen .
  • R 15 is hydrogen, methyl, ethyl, or -(CH 2 ) n CH 2 -R 25 wherein n is 0, 1, or 2; and R 25 is OH, OMe, F, CN, CF 3 , SOCH 3 or SO 2 CH 3 , -NR 30 COR 31 , -NR 30 COOR 31 , -NR 30 SO 2 R 31 , C(O)NR 30 R 31 , or has the formula:
  • R 33 , R 34 , R 35 , R 36 , and R 37 are hydrogen, alkyl, or substituted alkyl.
  • R 15 is ethyl, methoxyethyl, CH 2 CH 2 F, or CH 2 CH 2 CN;
  • R 17 is bromo or chloro;
  • R 18 is methoxy or hydrogen; and
  • R 19 is hydrogen.
  • R 15 is -(CH 2 )nCH 2 R 25 n is 0, 1, 2 or 3 and R 25 is OH, OMe, F, CN, CF 3 , SOCH 3 or SO 2 CH 3 , -NR 30 COOR 31 , - NR 30 C(O)R 31 , -NR 30 SO 2 R 31 , -C(O)NR 31 R 32 or has the formula:
  • R 33 , R 34 , R 35 , R 36 , and R 37 are hydrogen, alkyl, or substituted alkyl. More preferably, R 15 is methyl, ethyl, CH 2 F, methoxyethyl, CH 2 CH 2 F, or
  • R 17 is bromo
  • R 18 is hydrogen, methoxy, or fluoro
  • R 19 is H.
  • Y is selected from the group consisting of O and S;
  • W is selected from the group consisting of N, C, O, and S; provided that if W is O or S, R 9 is absent;
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 8 , R 9 are each independently selected from the group consisting of H, C 1- alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, -OR 60 , -NO 2 , -OH, -SR 60 , -NR 60 R 61 , -CN, -CO 2 R 60 , -CONR 60 R 61 , OCONR 60 R 61 ,
  • n 1 to 3; and Z is selected from the group consisting of - C 4 alkyl, alkenyl, and alkynyl chain; Z having one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR 60 SO 2 R 61 groups;
  • R is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, - NR 30 COOR 31 , -NR 30 C(O)R 31 , -NR 30 SO 2 R 31 , -C(O)NR 30 R 31 , heteroaryl or heterocycloalkyl; and
  • R 30 and R 31 are, independently, hydrogen, alkyl, or cycloalkyl.
  • tyrosine kinase inhibitors can enhance the activity of cytotoxic or cytostatic treatments when used in combination with standard therapies known in the art.
  • the invention also provides a pharmaceutical composition comprising a compound of formula I, as defined above, and a pharmaceutically acceptable carrier.
  • the invention further provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of formula I, as defined above, in combination with pharmaceutically acceptable carrier and at least one other anti-cancer agent optionally formulated as a fixed dose.
  • a method of treating a condition associated with at least one tyrosine kinase enzyme comprising administering to a mammalian species in need of such treatment an effective amount of a compound of formula I, as defined above.
  • the invention provides a method of treating a condition associated with at least one tyrosine kinase enzyme comprising administering to a mammalian species at least one other anti-cancer agent in combination with a compound of formula I, as defined above.
  • alkyl herein alone or as part of another group refers to a monovalent alkane (hydrocarbon) derived radical containing from 1 to 12 carbon atoms unless otherwise defined.
  • An alkyl group is an optionally substituted straight, branched or cyclic saturated hydrocarbon group. When substituted, alkyl groups may be substituted with up to four substituent groups, R as defined, at any available point of attachment.
  • alkyl group When the alkyl group is said to be substituted with an alkyl group, this is used interchangeably with "branched alkyl group".
  • branched alkyl group Exemplary unsubstituted such groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, and the like.
  • substituents may include but are not limited to one or more of the following groups: hydroxy, halo (such as F, CI, Br, I), haloalkyl (such as CCI3 or CF3), alkoxy, alkylthio, cyano, carboxy (-COOH), alkylcarbonyl (-C(O)R), alkoxycarbonyl (-OCOR), amino, carbamoyl (-NHCOOR or -OCONHR), urea (-NHCONHR), thiol, (-SH), sulfoxy, sulfonyl, aryl, heteroaryl, and heterocycloalkyl.
  • Alkyl groups as defined may also comprise one or more carbon to carbon double bonds or one or more carbon to carbon triple bonds.
  • Alkyl groups may also be represented by the formula alkyl-R 25 .
  • the alkyl group is a methyl, ethyl, propyl or butyl group and include substituted methyl, ethyl, propyl or butyl groups.
  • alkenyl herein alone or as part of another group refers to a hydrocarbon radical straight, branched or cyclic containing from 2 to 12 carbon atoms and at least one carbon to carbon double bond.
  • An alkenyl group may be optionally substituted in the same manner as described for an alkyl group.
  • alkynyl herein alone or as part of another group refers to a hydrocarbon radical straight, branched or cyclic containing from 2 to 12 carbon atoms and at least one carbon to carbon triple bond.
  • An alkynyl group may be optionally substituted in the same manner as described for an alkyl group.
  • alkoxy refers to a straight or branched chain alkyl group covalently bonded to the parent molecule through an oxygen atom linkage containing from one to ten carbon atoms and the terms "C 1-6 alkoxy” and “lower alkoxy” refer to such groups containing from one to six carbon atoms, examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy and the like.
  • alkoxy substituent when used in connection with an alkoxy substituent refers to the replacement of up to two hydrogens, preferably on different carbon atoms with a radical selected form the group of lower alkyl, phenyl, cyano, halo, u ⁇ fluoromethyl, nitro, hydroxy, alkanoyl, amino, monoalkyl amino and dialkylamino.
  • Alkoxy groups may be substituted in the same manner that alkyl groups can be substituted as described above.
  • sulfoxy herein alone or as part of a group refers to -SO and may be substituted with, for example, alkyl or aryl groups.
  • sulfonyl herein alone or as part of a group refers to -SO 2 and may be substituted with alkyl or aryl groups.
  • amino herein alone or as part of another group refers to -NH 2 .
  • An “amino” may optionally be substituted with one or two substituents, which may be the same or different, such as alkyl, aryl, arylalkyl, alkenyl, alkynyl, heteroaryl, heteroarylalkyl, cycloheteroalkyl, cycloheteroalkylalkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl or thioalkyl.
  • substituents include alkylamino and dialkylamino, such as methylamino, ethyl amino, dimethylamino, and diethylamino. These substituents may be further substituted with a carboxylic acid or any of the alkyl or aryl substituents set out herein.
  • amino substituents may be taken together with the nitrogen atom to which they are attached to form 1- pyrrolidinyl, 1-piperidinyl, 1-azepinyl, 4-morpholinyl, 4-thiamorpholinyl, 4- sulfoxymorpholine, 4-sulfonylmorpholine, 1-piperazinyl, 4-alkyl-l-piperazinyl, 4- arylalkyl-1-piperazinyl, 4-diarylalkyl- 1-piperazinyl , 1-homopiperazinyl, 4-alkyl-l- homopiperazinyl, 4-arylalkyl- 1-homopiperazinyl, 4-diarylalkyl-l-homopiperazinyl; 1- pyrrolidinyl, 1-piperidinyl, or 1-azepinyl, optionally substituted with alkyl, alkoxy, alkylthio, halo, trifluoromethyl or hydroxy .
  • aryl herein alone or as part of another group refers to monocyclic or bicyclic aromatic rings, e.g. phenyl, substituted phenyl and the like, as well as groups which are fused, e.g., napthyl, phenanthrenyl and the like.
  • An aryl group thus contains at least one ring having at least 6 atoms, with up to five such rings being present, containing up to 22 atoms therein, with alternating (resonating) double bonds between adjacent carbon atoms or suitable heteroatoms.
  • Aryl groups may also be substituted with heterocycloalkyl and heterocycloaryl groups to form fused rings, such as dihydiobenzfuranyl, oxindolyl, indolyl, indolinyl, oxindolyl, benzoxazolidinonyl, benzoxazolinyl and benzoxazolidinone.
  • cycloalkyl herein alone or as part of another group refers to fully saturated and partially unsaturated hydrocarbon rings of 3 to 9, preferably 3 to 7 carbon atoms. Further, a cycloalkyl may be substituted.
  • R' and R" is independently selected from hydrogen, alkyl, substituted alkyl, and cycloalkyl, or R' and R" together form a heterocyclo or heteroaryl ring.
  • Cycloalkyl groups may also be substituted with hetero atoms such as O, N, and S to form heterocycloalkyl groups.
  • Preferred heterocycloalkyl groups include optionally substituted morpholine, homomorpholine (7 membered ring), thiomorpholine, piperazine, homopiperazine (7 membered ring), and piperidine.
  • heteroaryl herein alone or as part of another group refers to substituted and unsubstituted aromatic 5 or 6 membered monocyclic groups, 9 or 10 membered bicyclic groups, and 11 to 14 membered tricyclic groups which have at least one heteroatom (O, S or N) in at least one of the rings.
  • Each ring of the heteroaryl group containing a heteroatom can contain one or two oxygen or sulfur atoms and/or from one to four nitrogen atoms provided that the total number of heteroatoms in each ring is four or less and each ring has at least one carbon atom.
  • the fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or unsaturated.
  • the nitrogen and sulfur atoms may optionally be oxidized and the nitrogen atoms may optionally be quatemized.
  • Heteroaryl groups which are bicyclic or tricyclic must include at least one fully aromatic ring but the other fused ring or rings may be aromatic or non- aromatic.
  • the heteroaryl group may be attached at any available nitrogen or carbon atom of any ring.
  • Exemplary monocyclic heteroaryl groups include pyrrolyl, pyrrolidinyl, imidazolinyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, and the like.
  • Exemplary bicyclic heteroaryl groups include indolyl, indolinyl, oxindolyl, benzoxazolidinone, benzothiazolyl, benzodioxolyl, benzoxazolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, fiiropyridinyl, dihydroisoindolyl, tetrahydroquinolinyl and the like.
  • Exemplary tricyclic heteroaryl groups include carbazolyl, benzindolyl, phenanthrollinyl, acridinyl, phenanthridinyl, xanthenyl and the like.
  • halogen or halo
  • hydroxy herein alone or as part of another group refers to -OH.
  • thioalkoxy herein alone or as part of another group refers to an alkyl group as defined herein attached to the parent molecular group through a sulfur atom.
  • examples of thioalkoxy include, but are not limited to, thiomethoxy, thioethoxy, and the like.
  • an "anti-cancer agent” as used herein includes known anti-cancer treatments such as radiation therapy or with cytostatic or cytotoxic agents, such as for example, but not limited to, DNA interactive agents, such as cisplatin or doxorubicin; topoisomerase II inhibitors, such as etoposide; topoisomerase I inhibitors such as CPT-11 or topotecan; tubulin interacting agents, such as paclitaxel, docetaxel or the epothilones; hormonal agents, such as tamoxifen; thymidilate synthase inhibitors, such as 5-fluorouracil; anti-metabolites, such as methotrexate; tyrosine kinase inhibitors such as Iressa and OSI-774; angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDK inhibitors; Herl/2 inhibitors and monoclonal antibodies directed against growth factor receptors such as erbitux (EGF) and herceptin (Her2).
  • protecting groups for the compounds of the present invention will be recognized from the present application taking into account the level of skill in the art, and with reference to standard textbooks, such as Greene, T. W. et al., Protective Groups in Organic Synthesis, Wiley, N.Y. (1991).
  • alkenyl, alkynyl, cycloalkyl are substituted, they are preferably substituted with one or more hydroxy, cyano, carbamoyl, hydroxy, alkoxy, thiol, alkenyl, thioalkoxy, amino, alkylamino, amido, sulfonyl, sulfoxy, sulfonamido, halo, heterocycloalkyl, aryl or heteroaryl.
  • aryl or heteroaryl When aryl or heteroaryl are substituted, they are preferably substituted with one or more alkyl, alkenyl, alkynyl, cyano, carbamoyl, hydroxy, alkoxy, thioalkoxy, amino, amido, sulfonamido, halo or with R', R" wherein R', R" form a ring that is fused to the aryl group.
  • CH 2 aryl or heteroaryl When CH 2 aryl or heteroaryl are substituted, they are preferably substituted with one or more alkyl, alkyenyl, alkynyl, cyano, carbamoyl, hydroxy, alkoxy, thioalkoxy, amino, amido, sulfonamido, or halogen.
  • NH-Z-aryl or NH-Z-heteroaryl groups are substituted, they are preferably substituted with one or more alkyl, alkenyl, alkynyl, hydroxy, alkoxy, thioalkoxy, amino, halogen, nitro, nitrile, carboxylate, alkoxycarbonyl, carbamoyl, ester, amide, aryl, or heteroaryl
  • alkyl alkenyl, alkynyl, hydroxy, alkoxy, thioalkoxy, amino, halogen, nitro, nitrile, carboxylate, alkoxycarbonyl, carbamoyl, ester, amide, aryl, or heteroaryl
  • C ⁇ _ alkyl means a straight or branched saturated carbon chain having from one to six carbon atoms; examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t- butyl, n-pentyl, sec-pentyl, isopentyl, and n-hexyl.
  • Cj.g alkyl can also refer to C . alkylene which bridges two groups; examples include propane- 1, 3 -diyl, butane- 1,4-diyl, 2-methyl-butane-l,4-diyl, etc.
  • C2-6 alkenyl means a straight or branched carbon chain having at least one carbon-carbon double bond, and having from two to six carbon atoms; examples include ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, pentenyl, and hexenyl.
  • C2-6 alkenyl can also refer to C2-6 alkenediyl which bridges two groups; examples include ethylene-l,2-diyl (vinylene), 2-methyl-2-butene- 1,4-diyl, 2-hexene-l,6-diyl, etc.
  • C2-6 alkynyl means a straight or branched carbon chain having at least one carbon-carbon triple bond, and from two to six carbon atoms; examples include ethynyl, propynyl, butynyl, and hexynyl.
  • alkyl-R 25 includes optionally substituted alkyl groups such as methyl, ethyl, propyl, and butyl, attached to an R 25 group.
  • R 25 generally includes hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, -NHCOOH, -NHC(O)-, -NHSO 2 -, -C(O)NH 2 , heteroaryl or heterocycloalkyl groups such as morpholinyl or group having the formula:
  • imidazole and imidazoline herein alone or as part of another group includes substituted imidazoles and substituted imidazolines.
  • tetrahydropyrimidine includes substituted tetrahydropyrimidines.
  • piperazine include substituted piperazines, substituted piperidines, substituted mo ⁇ holines, substituted homomo ⁇ holines and substituted pyrrolidines, respectively.
  • X is selected from the group consisting of N, C, C ⁇ -C 3 alkyl, -C 3 alkyl substituted with one or more R 7 , and a direct bond;
  • Y is selected from the group consisting of O and S ;
  • W is selected from the group consisting of N, C, O, and S, provided that when W is O or S, R 9 is absent;
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 are each independently selected from the group consisting of H, C 1-6 alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, OR 60 , NO 2 , OH, SR 60 , NR 60 R 61 , CN, CO 2 R 60 , CONR 60 R 61 , CO 2 NR 60 R 61 , NR 62 CONR 60 R 61 , NR 60 SO 2 R 61 , SO 2 NR 60 R 61 , C(NR 62 )NR 60 R 61 , aryl, heteroaryl, (CH 2 ) n OR 60 , (CH 2 ) n NR 60 R 61 , (CH 2 ) n SR 60 , (CH 2 ) n aryl, (CH 2 ) n heteroaryl, , (CH 2
  • Z is selected from the group consisting of - C 4 alkyl, alkenyl, and alkynyl chain; Z having one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR 60 SO 2 R groups; Z optionally inco ⁇ orating one or more groups selected from the group consisting of CO, CNOH, CNOR 60 , CNNR 60 , CNNCOR 60 and CNNSO 2 R 60 ; and R 60 , R 61 , R 62 and R 63 are independently selected from the group consisting of
  • alkyl alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy, alkoxy, aryl, heteroaryl, heteroarylalkyl, and alkyl-R wherein
  • R 25 is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, - NR 30 COOR 31 , -NR 30 C(O)R 31 , -NR 30 SO 2 R 31 , -C(O)NR 30 R 31 , heteroaryl or heterocycloalkyl; and
  • R and R are, independently, hydrogen, alkyl, or cycloalkyl.
  • R 3 is -OR 60 .
  • R 60 is alkyl, or - alkyl-R , wherein R is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, cyano, alkylsulfoxy, alkylsulfonyl, -R 30 COOR 31 , -NR 30 C(O)R 31 , - NR 30 SO 2 R 31 , -C(O)NR 30 R 31 , heteroaryl or heterocycloalkyl; and R 30 and R 3l are, independently, hydrogen, alkyl, or cycloalkyl.
  • R 60 is methyl, -(CH 2 ) n CH 2 OH, or -(CH 2 ) n CH 2 N(CH 2 CH 2 ) 2 O, and n is 0, 1, or 2.
  • R is piperazine, homopiperazine, 3-methylpiperazine, or 3,5-dimethylpiperazine being optionally substituted at the 4-N position with a compound selected from the group consisting of alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, alkyl-R 25 , -C(O)-R 15 , or -CO 2 R 15 wherein R 15 is hydrogen, alkyl, aryl, alkyl-R 25 , amino or aryl; and R 25 is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, cyano, halo, sulfoxy, sulfonyl, arylsulfonyl, -NR 30 COOR 31 , -NR 30 C(O)R 31 , -NR 30 SO 2 R 31 , -C(O)NR 30 R
  • piperazine is substituted with Me, CH 2 cyclopropyl, CH 2 CH 2 NMe 2 , CH 2 CH 2 NEt 2 , CH 2 CH 2 NH 2 , CH 2 CH 2 NHMe, CH 2 CH 2 NHEt, N-CH 2 CH 2 N(CH 2 CH 2 ) 2 O, (CH 2 ) n CH 2 -R 25 wherein R 25 is OH, OMe, F, CN, CF 3 , SOCH 3 or SO 2 CH 3 , wherein n is 0, 1 , or 2.
  • R 3 is an amino group.
  • Preferred amino groups include NHCH 2 CH 2 OH, NMeCH 2 CH 2 OH, NEtCH 2 CH 2 OH, NHCH 2 CH 2 NH 2 , NMeCH 2 CH2NH 2 , NEtCH 2 CH 2 NH 2 , NHCH 2 CH 2 NMe 2 , NMeCH 2 CH 2 NMe 2 , NEtCH 2 CH 2 NMe2, NHCH 2 CH 2 NEt 2 , NMeCH 2 CH2NEt 2 , NEtCH 2 CH 2 NEt 2 , NHCH2CH2N(CH2CH 2 )2 ⁇ , NMeCH2CH2N(CH 2 CH2) 2 ⁇ , NEtCH 2 CH 2 N(CH2CH2) 2 ⁇ .
  • R 3 is an optionally substitued piperidine.
  • Preferred substituents are selected from the group consisting of hydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, 1,3 dioxolane (-OCHR 15 ) 2 , 1,3 dioxane (-OCHR 15 CHR 15 CHR 15 O-) -NHC(O)R 15 , -NHCO 2 R 15 , wherein R 15 is hydrogen, alkyl or alkyl-R 25 .
  • R is an optionally substituted mo ⁇ holine, homomo ⁇ holine, thiomo ⁇ holine, sulfoxymo ⁇ holine, or sulfonylmo ⁇ holine.
  • Preferred substituents include hydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, alkyl-R 25 , -NHC(O)R 15 , -NHCO 2 R 15 , wherein R 15 is hydrogen, alkyl or alkyl-R 25 wherein R 25 .
  • R 3 is a pyrrolidine.
  • Preferred pyrrolidines include, 3- hydroxyl pyrrolidine, 3- alkoxy pyrrolidine, and 3-alkylamino pyrrolidine.
  • R 3 is an optionally substituted N-tetrahydropyrimidine or N-imidazoline wherein the substituents are, preferably, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cyanoalkyl, carboxyl, or carboxamide.
  • R 6 is is selected from the group consisting of H, 2- aminomethylpyridine, NHCH 2 CH(OH)aryl, and NHCH(CH 2 OH)CH 2 aryl, wherein the aryl group is optionally substituted.
  • the aryl group is substituted with Br, CI, F, CN, or methoxy.
  • R 6 has one of the following formulae:
  • R 40 is hydrogen or alkyl, preferably methyl, and R 17 is hydrogen or halogen, such as Br, CI or F.
  • Preferred compounds of the present invention have one of the following formulae:
  • R 12 and R 13 are, independently, hydrogen, alkyl, or alkyl-R 25 ;
  • R 15 is hydrogen, alkyl, or alkyl-R 25 ;
  • R 16 is independently, hydrogen or methyl
  • R 17 , R 18 and R 19 are, independently, hydrogen, halogen, or alkoxy,or R 18 and
  • R .19. together form a heterocycloalkyl or heteroaryl group
  • R 25 is hydrogen, cycloalkyl, hydroxy, thiol, alkenyl, alkoxy, thioalkoxy, amino, halo, cyano, sulfoxy, sulfonyl, -NR 30 COOR 31 , -NR 30 C(O)R 31 , -NR 30 SO 2 R 31 ,
  • R ,30 and R > 31 are, independently, hydrogen, alkyl, or cycloalkyl.
  • R 12 is hydrogen, methyl, hydroxymethyl, methoxymethyl, alkyl-R 25 , CH 2 F, CH CN, CO 2 H, or -CONR 30 R 31 ;
  • R 13 is H;
  • R 17 is Br, Cl or F;
  • R R is halo or methoxy
  • R 19 is H.
  • Y is selected from the group consisting of O and S;
  • W is selected from the group consisting of N, C, O, and S; provided that if W is O or S, R 9 is absent;
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 8 , R 9 are each independently selected from the group consisting of H, C 1-6 alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, -OR 60 , -NO 2 , -OH, -SR 60 , -NR 60 R 61 , -CN, -CO 2 R 60 , -CONR 60 R 61 , OCONR 60 R 61 ,
  • n 1 to 3; and Z is selected from the group consisting of C ⁇ - C 4 alkyl, alkenyl, and alkynyl chain; Z having one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR 60 SO 2 R 61 groups
  • R is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, - NR 30 COOR 31 , -NR 30 C(O)R 31 , -NR 30 SO 2 R 31 , -C(O)NR 30 R 31 , heteroaryl or heterocycloalkyl; and
  • R and R are, independently, hydrogen, alkyl, or cycloalkyl.
  • R 1 is -CO 2
  • R 60 is -CO 2
  • R 2 , R 4 ,R 5 and R 8 are H or methyl
  • R 3 is a heterocycloalkyl
  • R 6 is -NH(CH) 2 ⁇ H-aryl.
  • Preferred heterocycles include mo ⁇ holine or homomo ⁇ holine.
  • Suitable examples of salts of the compounds according to the invention include inorganic or organic acids. These include, but are not limited to, hydrochloride, hydrobromide, sulfate, methanesulfonate, maleate, fumarate, phosphate and other pharmaceutically acceptable salts. Salts which are unsuitable for pharmaceutical uses but which can be employed, for example, for the isolation or purification of free compounds of formula I or their pharmaceutically acceptable salts, are also included.
  • All stereoisomers of the compounds of the instant invention are contemplated, either in admixture or in pure or substantially pure form.
  • the definition of the compounds according to the invention embraces all possible stereoisomers and their mixtures. It very particularly embraces the racemic forms and the isolated optical isomers having the specified activity.
  • the racemic forms can be resolved by physical methods, such as, for example, fractional crystallization, separation or crystallization of diastereomeric derivatives or separation by chiral column chromatography.
  • the individual optical isomers can be obtained from the racemates by conventional methods, such as, for example, salt formation with an optically active acid followed by crystallization.
  • prodrug forms of the compounds of formula I includes prodrug forms of the compounds of formula I.
  • Various forms of prodrugs are well known in the art.
  • prodrug derivatives see: (a) Design of Prodrugs, edited by H. Bundgaard (Elsevier, 1985); and Methods in Enzymology, Vol. 42, pp. 309-396, edited by K.
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of formula I, as defined above, and a pharmaceutically acceptable carrier and at least one other anti-cancer agent formulated as a fixed dose.
  • Preferred anti- cancer agents are selected from the group consisting of: tamoxifen, toremifen, raloxifene, droloxifene, iodoxyfene, megestrol acetate, anastrozole, letrazole, borazole, exemestane, flutamide, nilutamide, bicalutamide, cyproterone acetate, goserelin acetate, luprolide, finasteride, herceptin, methotrexate, 5-fluorouracil, cytosine arabinoside, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin, cisplatin, carboplatin, melphalan, chlorambucil, busulphan, cyclophosphamide, ifosfamide, nitrosoureas, thiotephan
  • the invention provides a method of treating a condition via modulation of at least one tyrosine kinase enzyme comprising administering to a mammalian species in need of such treatment an effective amount of a compound of formula I, as defined above, in combination (simultaneously or sequentially) with at least one other anti-cancer agent.
  • a preferred condition, treated by said methods of the instant invention, is cancer.
  • the tyrosine kinase enzyme may include (but is not limited to): Abl, CDK's, EGF, EMT, FGF, FAK, Flk-1/KDR, HER-2, IGF-1R, IR, LCK, MET, PDGF, Src, and VEGF.
  • the invention also provides a method for treating cancer, comprising administering to a mammalian species in need of such treatment, a therapeutically effective amount of at least one of the pharmaceutical compositions defined above.
  • the invention further provides a method for treating proliferative diseases, comprising administering to a mammalian species in need of such treatment a therapeutically effective amount of at least one of the pharmaceutical compositions defined above.
  • Certain compounds of formula I may generally be prepared according to the following schemes and the knowledge of one skilled in the art. Solvates (e.g., hydrates) of the compounds of formula I are also within the scope of the present invention. Methods of solvation are generally known in the art. Accordingly, the compounds of the instant invention may be in the free or hydrate form, and may be obtained by methods exemplified by the following schemes below.
  • Schemes I-V ⁇ illustrate the preparation of compounds claimed in this invention.
  • the examples, which follow, illustrate the compounds that can be synthesized by these schemes. The schemes are not limited by the examples listed or by any substituents employed for illustrative pu ⁇ oses.
  • Scheme I describes the preparation of the benzimidazoles.
  • the starting diamines 1 are readily available using literature methods or are obtained commercially.
  • the diamine is then condensed with an aldehyde 2 to provide the benzimidazole 3. Further modification of the functional groups on the aryl group of the benzimidazole or heterocycle of 3 are then envisioned.
  • the benzimidazole could be formed in a step-wise manner (see Scheme H) by amide formation using the acid chloride of 5 or any of the commonly used peptide coupling reagents such as DCC (dicyclohexylcarbodiimide), EDCI (l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), etc.
  • DCC dicyclohexylcarbodiimide
  • EDCI l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
  • the amide 6 was formed the nitro group could be reduced using catalytic hydrogenation, transfer hydrogenation or chemical reduction such as SnC ⁇ or iron powder or other methods known in the art for reduction of aryl nitro groups. Treatment of the aniline with acid would then form the benzimidazole.
  • Scheme HI illustrates the use of 4-iodo-2-methoxy-pyridine-3- carbaldehyde 7 to provide the functionalized benzimidazole 8.
  • Hydrolysis of the methoxy group using protic acid conditions, TMSI (trimethylsilyl iodide), BBr 3 , or other conditions known in the art for cleaving a methyl ether would provide the halopyridone 9.
  • Addition of heteroatom nucleophiles using amines, alcohols or thiols would then provide the substituted pyridones 10.
  • Other functionality could be inco ⁇ orated into the aldehyde and the above example is included for illustrative pu ⁇ oses only.
  • aryl ring of the benzimidazole prepared using Schemes I or II can be modified.
  • introduction of a cyano group for R 3 on the benzimidazole allows for the formation of heterocycles such as imidazole, imidazolines, oxazolines, thiazolines, amides, or amidines.
  • Scheme IV illustrates such transformations. Starting from the cyano-substituted benzimidazole 11 the heterocycle can be modified as illustrated in Scheme IV to provide 12. Imidate formation preferably using ethanol and acid provides intermediate 13.
  • Imidate 13 can be transformed using diamines to form imidazolines, amino alcohols to form oxazolines, amino acetals to form imidazoles, and amino thiols to form thiazolines 14.
  • the imidate can be hydrolyzed to the acid and coupled with amines using any of the standard amide formation reagents (DCC, EDCI, etc.) to form amides 15.
  • Imidate 13 is also a useful intermediate for the preparation of amidines 16 by reacting with amines.
  • Scheme V illustrates further transformation of benzimidazoles that bear a halogen atom using palladium catalysis using conditions developed by Suzuki [Yang et al. Acta Chem. Scand. (1993) 221; Suzuki et al. Synth. Commun. (1981) 11: 513] or Buchwald/Hartwig [Buchwald et al. J. Am. Chem. Soc. (1994) 116: 7901; Hartwig et al. J. Am. Chem. Soc. (1994) 116: 5969; Hartwig. Angew. Chem., Int. Ed. Engl. (1998) 37: 2046] and variations of these methods.
  • bromide substituted benzimidazole 17 could be envisioned to provide a substrate for Suzuki coupling with aryl, vinyl, and heterocyclic boronic acids to provide benzimidazoles 18.
  • amines and heterocycles such as piperazine or mo ⁇ holine derivatives 19 can be prepared from the same bromide using amines under conditions described by Buchwald and Hartwig or variations thereof.
  • amine and heterocyclic derivatives such as 19 can be prepared using intermediate 6 described in Scheme II.
  • the halogen can be displaced with amines, alcohols, heterocyclic amines and other nitrogen containing heterocycles such as piperazine, piperidine, 4-amino piperidine, mo ⁇ holine, imidazole, etc (Scheme VI).
  • the terminal nitrogen of piperazine or 4-amino piperidine can then be alkylated using standard alkylation conditions or reacted with aldehydes in a reductive amination reaction to provide alkylated derivatives.
  • terminal nitrogen atom of piperazine or 4- amino piperidine can be acylated or carbamoylated using any number of conditions that are routine for someone skilled in the art of organic synthesis.
  • Scheme JJ compounds such as 19 could be prepared.
  • amines, heterocycles, and alcohols can be introduced at R 3 using a nucleophilic aromatic substitution reaction started from an intermediate 20 were R 3 is halogen, preferably F, the halogen can be displaced with amines, alcohols, heterocyclic amines and other nitrogen containing heterocycles such as piperazine, piperidine, 4-amino piperidine, mo ⁇ holine, imidazole, etc (Scheme VH).
  • the terminal nitrogen of piperazine or 4-amino piperidine can then be alkylated using standard alkylation conditions or reacted with aldehydes in a reductive animation reaction to provide alkylated derivatives.
  • the terminal nitrogen atom of piperazine or 4-amino piperidine can be acylated or carbamoylated using any number of conditions that are routine for someone skilled in the art of organic synthesis.
  • the resulting nitro aniline could be reduced to the diamine 21 and processed as illustrated in Scheme DI.
  • Cyclization may then be performed using a base such as sodium hydride, potassium tert-butoxide, lithium bis(trimethylsilyl)amide or the like to give compounds 26 which will then be converted to the corresponding sulfonates 27 by reaction with various alkylsulfonyl halides or alkylsulfonic anhydrides such as methanesulfonyl chloride, p- toluenesulfonyl chloride or preferably trifluoromethanesulfonic anhydride in presence of a weak base such as pyridine, 2,6-di-tert-butyl-4-methyl-pyridine or triethylamine. Substitution with heteroatom nucleophiles such as amines, thiols or alcohols provides the desired 5-, 6- or 7-membered cyclic cores 28.
  • a base such as sodium hydride, potassium tert-butoxide, lithium bis(trimethylsilyl)amide or the like to give compounds 26
  • the compounds according to the invention have pharmacological properties; in particular, the compounds of formula I are tyrosine kinase enzyme inhibitors.
  • the novel compounds of formula I are thus useful in the therapy of a variety of proliferative diseases (including but not limited to diseases associated with tyrosine kinase enzymes) such as cancer, autoimmune diseases, viral diseases, fungal diseases, neurodegenerative disorders and cardiovascular disease.
  • the compounds of formula I are useful in the treatment of a variety of cancers, including, but not limited to, the following: a) carcinoma, including that of the bladder, breast, colon, kidney, liver, lung, including small cell lung cancer, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, including squamous cell carcinoma; b) hematopoietic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma and Burkett's lymphoma; c) hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias, myelodysplastic syndrome and promyelocytic leukemia; d) tumors of me
  • inhibitors could act as reversible cytostatic agents which may be useful in the treatment of any disease process which features abnormal cellular proliferation, e.g., benign prostatic hype ⁇ lasia, familial adenomatosis polyposis, neuro-fibromatosis, atherosclerosis, pulmonary f ⁇ brosis, arthritis, psoriasis, glomerulonephritis, restenosis following angioplasty or vascular surgery, hypertrophic scar formation, inflammatory bowel disease, transplantation rejection, endotoxic shock, and fungal infections.
  • Compounds of formula I may induce apoptosis.
  • apoptotic response is aberrant in a variety of human diseases.
  • Compounds of formula I, as modulators of apoptosis, will be useful in the treatment of cancer (including but not limited to those types mentioned herein above), viral infections (including but not limited to he ⁇ esvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus), prevention of AIDS development in HlV-infected individuals, autoimmune diseases (including but not limited to systemic lupus, erythematosus, autoimmune mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes mellitus), neurodegenerative disorders (including but not limited to Alzheimer's disease, AIDS -related dementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, spinal muscular atrophy and cerebellar degeneration), myelodysplastic syndromes, aplastic anemia,
  • Compounds of formula I may modulate the level of cellular RNA and DNA synthesis. These agents would therefore be useful in the treatment of viral infections (including but not limited to HIV, human papilloma virus, he ⁇ esvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus).
  • viral infections including but not limited to HIV, human papilloma virus, he ⁇ esvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus.
  • Chemoprevention is defined as inhibiting the development of invasive cancer by either blocking the initiating mutagenic event or by blocking the progression of pre- malignant cells that have already suffered an insult or inhibiting tumor relapse.
  • Compounds of formula I may also be useful in inhibiting tumor angiogenesis and metastasis.
  • the compounds of this invention may also be useful in combination (administered together or sequentially) with known anti-cancer treatments such as radiation therapy or with cytostatic or cytotoxic agents, such as for example, but not limited to, DNA interactive agents, such as cisplatin or doxorubicin; topoisomerase II inhibitors, such as etoposide; topoisomerase I inhibitors such as CPT-11 or topotecan; tubulin interacting agents, such as paclitaxel, docetaxel or the epothilones; hormonal agents, such as tamoxifen; thymidilate synthase inhibitors, such as 5-fluorouracil; and anti-metabolites, such as methotrexate; tyrosine kinase inhibitors such as Iressa and OSI-774 (Tarceva 1 " 1 ) ; Avastin, Herceptin, angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDK inhibitors; Herl/2 inhibitors
  • Such combination products employ the compounds of this invention within the dosage range described below and the other pharmaceutically active agent or treatment within its approved dosage range.
  • Compounds of formula I may also be administered sequentially with known anticancer or cytotoxic agents when a combination formulation is inappropriate. The invention is not limited in the sequence of administration; compounds of formula I may be administered either prior to or after administration of the known anticancer or cytotoxic agent(s).
  • compositions for use as described above, including controlling cancer, inflammation and arthritis, which contain at least one compound of the formula I as defined above or at least one of its pharmacologically acceptable acid addition salts, and the use of a compound of the formula I as defined above for the preparation of a pharmaceutical having activity against proliferative diseases as described previously including against cancer, inflammation and/or arthritis.
  • the pharmacological properties of the compounds of this invention may be confirmed by a number of pharmacological assays.
  • the exemplified pharmacological assays which follow have been carried out with the compounds according to the invention and their salts.
  • kinase reactions consisted of 5 ng of baculovirus expressed GST- CDK2/cyclin E complex, 0.5 ⁇ g GST-RB fusion protein (amino acids 776-928 of retinoblastoma protein), 0.2 ⁇ Ci 33 P ⁇ -ATP, 25 ⁇ M ATP in 50 ⁇ l kinase buffer (50 mM Hepes, pH 8.0, 10 mM MgCl 2) 1 mM EGTA, 2 mM DTT). Reactions were incubated for 45 minutes at 30° C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration of 15%.
  • TCA cold trichloroacetic acid
  • TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT).
  • Dose response curves were generated to determine the concentration required inhibiting 50% of kinase activity (IC 50 ).
  • Compounds were dissolved at 10 mM in dimethylsulfoxide (DMSO) and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equaled 2%.
  • a filter-based kinase assay measuring the phosphorylation of Gst-SLP76 by Gst-Emtk, was employed to determine the compound inhibitory activity against Emt.
  • the kinase reaction was performed in a 96-well plate at room temperature for 15 min before being terminated by adding 100 ⁇ l of 20% trichloroacetic acid (TCA) containing 0.1 M sodium pyrophosphate.
  • TCA trichloroacetic acid
  • the kinase reaction mixture (60 ⁇ l ) contained 25 mM HEPES, pH 7.0, 0.1 mg/ml BSA, 5 mM MgCl 2 , 5 mM MnCl 2 , 8 ng of enzyme (Gst-Emtk), 5 ⁇ g of the substrate protein (Gst-SLP76), 1 ⁇ M ATP, 0.4 ⁇ Ci of [ ⁇ -P 33 ] ATP and the tested compound (at various concentrations). After termination, the proteins were allowed to precipitate in the presence of TCA for 1 hr at 4 °C.
  • the precipitated proteins were then harvested on a filter plate (UniFilter-96, GF/C, Packard Instrument) and washed to remove excess [ ⁇ -P 33 ]ATP.
  • the radioactivity was determined using a TopCount NXT (Packard Instrument) after adding 35 ⁇ l of Microscint 20 (Packard instrument).
  • the Focal Adhesion kinase was assayed using the synthetic polymer poly(Glu/Tyr) (Sigma Chemicals) as a phosphoacceptor substrate.
  • Each reaction mixture consisted of a total volume of 50 ul and contained 100 ng of baculovirus- expressed enzyme, 2 ⁇ g of ⁇ oly(Glu/Tyr), l ⁇ M of ATP, and 0.2 ⁇ Ci of [ ⁇ - 33 P]ATP.
  • the mixtures also contained 40 mM Tris.HCl, pH 7.4, 1 mM MnCl 2 , 0.5 mM DTT, and 0.1 mg/ml bovine serum albumin.
  • kinase activity was determined by quantitation of the amount of radioactive phosphate transferred to the poly(Glu/Tyr) substrate.
  • Inco ⁇ oration was measured by the addition of cold trichloroacetic acid (TCA) precipitation of the proteins which were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT).
  • TCA cold trichloroacetic acid
  • kinase reactions consisted of 10 ng of baculovirus expressed GST- HER1, 100 ng of HER2, 100 ng/ml poly(Glu/Tyr) (Sigma), 0.2 ⁇ Ci 33P ⁇ -ATP, 1 ⁇ M ATP in 50 ⁇ l kinase buffer (50 mM Tris, pH 7.5, 10 mM MnC12, 0.5 mM DTT). Reactions were incubated for lh at 27 C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration 15%.
  • TCA cold trichloroacetic acid
  • TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co.,
  • IC 50 concentration required to inhibit 50% of kinase activity
  • Compounds were dissolved at 10 mM in dimethylsulfoxide (DMSO) and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equaled 1%.
  • IGF- Receptor Tyrosine Kinase Assay The IGF-1 receptor tyrosine kinase was assayed using the synthetic polymer poly(Glu Tyr) (Sigma Chemicals) as a phosphoacceptor substrate. Each reaction mixture consisted of a total volume of 50 ul and contained 125 ng of baculovirus expressed enzyme, 2.5 ⁇ g of poly(Glu/Tyr), 25 ⁇ M of ATP, and 0.1 ⁇ Ci of [ ⁇ - 33 P]ATP. The mixtures also contained 20 mM MOPS, pH 7.0, 5 mM MnCl 2 , 0.5 mM DDT, and 0.1 mg/ml bovine serum albumin. The reaction mixtures were incubated at
  • kinase activity was determined by quantitation of the amount of radioactive phosphate transferred to the poly(Glu/Tyr) substrate.
  • Inco ⁇ oration was measured by the addition of cold trichloroacetic acid (TCA) precipitation of the proteins which were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT).
  • TCA cold trichloroacetic acid
  • the Insulin Receptor Tryrosine kinase was assayed using the synthetic polymer poly(Glu/Tyr) (Sigma Chemicals) as a phosphoacceptor substrate.
  • Each reaction mixture consisted of a total volume of 50 ul and contained 90 ng of baculovirus-expressed enzyme, 2.5 ⁇ g of poly(Glu/Tyr), 25 ⁇ M of ATP, and 0.1 ⁇ Ci of [ ⁇ - 33 P]ATP.
  • the mixtures contained also 20 mM Tris.HCl, pH 7.4, 5 mM MnCl 2 , 0.5 mM DTT, and 0.1 mg/ml bovine serum.
  • kinase activity was determined by quantitation of the amount of radioactive phosphate transferred to the poly(Glu/Tyr) substrate.
  • Inco ⁇ oration was measured by the addition of cold trichloroacetic acid (TCA) precipitation of the proteins which were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT).
  • TCA cold trichloroacetic acid
  • kinase reactions consisted of 10 ng of baculovirus expressed lOng GST-Lck, 100 ng/ml poly(Glu/Tyr) (Sigma), 0.2 ⁇ Ci 33P ⁇ -ATP, 1 ⁇ M ATP in 50 ⁇ l kinase buffer (50 mM Tris, pH 7.5, 10 mM MnC12, 0.5 mM DTT). Reactions were incubated for lh at 27 C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration 15%.
  • TCA cold trichloroacetic acid
  • TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co.,
  • IC 5 Q concentration required to inhibit 50% of kinase activity
  • Compounds were dissolved at 10 mM in dimethyl sulfoxide (DMSO) and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equaled 1%.
  • MET Kinase Assay Kinase reactions consisted of lOng of baculovirus expressed GST-Met, 2.5ug poly(Glu/Tyr) (Sigma), 0.2 ⁇ Ci 33P ⁇ -ATP, 10 ⁇ M ATP in 50 ⁇ l kinase buffer (40mM Tris, pH 7.5, ImM MnC12, 0.50 mM DTT). Reactions were incubated for lh at 27 C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration 3.5%.
  • TCA cold trichloroacetic acid
  • TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT).
  • Dose response curves were generated to determine the concentration required to inhibit 50% of kinase activity (IC 50 ).
  • Compounds were dissolved at 10 mM in dimethylsulfoxide (DMSO) and evaluated at seven concentrations, each in triplicate.
  • biotinylated poly(Glu/Tyr) (Sigma), in 50 ⁇ l kinase buffer (20 mM Hepes, pH 7.5, 0.7uM ATP, lOmM MnC12, 0.5mM DTT, 0.15mM NaCl, O.lmg/ml BSA).
  • Reactions were incubated for 30 minutes at room temperature with shaking and stopped by the addition of lOul of 0.2M EDTA, ⁇ H8.0. 150ul of HTRF detection buffer was added and incubated for 1 hour and 30 minutes at room temperature.
  • kinase reactions consisted of 7.5ng of baculovirus expressed GST-KDR, 1.5ug poly(Glu/Tyr) (Sigma), 0.04 ⁇ Ci 33P ⁇ -ATP, 2.5 ⁇ M ATP in 50 ⁇ l kinase buffer (25 mM Tris, pH 7.5, 1.8 mM MnC12, 0.0.625 mM DTT). Reactions were incubated for lh at 27 C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration 15%.
  • TCA cold trichloroacetic acid
  • TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co.,
  • IC 50 concentration required to inhibit 50% of kinase activity
  • Compounds were dissolved at 10 mM in dimethylsulfoxide (DMSO) and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equaled 1%.
  • K. Cytotoxicity assay (HT-29-colon; Colo205, MCF-7-breast)
  • Tumor cell lines are maintained in McCoy's 5 A medium (GIBCO) and 10% heat inactivated fetal bovine serum (GIBCO).
  • the in vitro cytotoxicity is assessed in tumor cells by a tetrazolium-based colorimetric assay which takes advantage of the metabolic conversion of MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulphenyl)-2H-tetrazolium, inner salt) (Promega) to a reduced form that absorbs light at 492 nm (1). Cells are seeded 24 hr prior to drug addition.
  • Biological Activity compounds of the present invention had kinase activity of ⁇ 25uM against one or more of the following kinases CDK, EMT, FAK, Herl, Her2, IGF, IR, LCK, MEK, MET, PDGF, VEGF.
  • HT-29 and Colo205 are human colon tumor cell lines, and MCF-7 is a human breast tumor cell line.
  • 5-Chloroacetyl-7-chlorooxindole To a suspension of A1C1 3 (13.4g, 0.10 mol) in methylene chloride (40mL) is added a solution of 7-Chlorooxindole (0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution is warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washing with saturated bicarbonate solution, brine, and drying over MgSO 4 would provide the desired chloroketone.
  • 6-Chloroacetyl-4-chloro-2-benzooxazolinone To a suspension of A1C1 3 (13.4g, 0.10 mol) in methylene chloride (40mL) is added a solution of 4-chloro-2- benzooxazolinone (0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution is warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washing with saturated bicarbonate solution, brine, and drying over MgSO 4 would provide the desired chloroketone.
  • N-Methyl-7-chloroindoline To a solution of the 7-Chloroindoline (0.10 mol) in 500 mL of acetone is added K 2 CO 3 (0.15mol) and Mel (0.15mol) and refluxed until the starting material is consumed. The reaction is filtered and washed with water and saturated bicarbonate solution, drying over MgSO 4 would provide the N-Me -7- chloroindoline
  • N-Methyl-5-chloroacetyl-7-chloro-indoline To a suspension of A1C1 3 (13.4g, 0.10 mol) in methylene chloride (40mL) is added a solution of N-Me-7-Chloroindoline (0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution is warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washing with saturated bicarbonate solution, brine, and drying over MgSO 4 provides the desired chloroketone.
  • 6-Chloroacetyl-4-chloro-2-methyl-benzooxazole To a suspension of A1C1 3 (13.4g, 0.10 mol) in methylene chloride (40mL) is added a solution of 2-Methyl-4-chloro- benzooxazole (0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution is warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washing with saturated bicarbonate solution, brine, and drying over MgSO 4 would provide the desired chloroketone.
  • N-(4-Imidazol-l-yl-2-methyl-6-nitro-phenyl)-acetamide To a solution of 4-imidazol- l-yl-2-methyl-phenylamine (1 g, 5.78 mmol) in CH 2 C1 2 (20 mL) was added Ac 2 O (0.7 mL, 7.28 mmol) at 0°C. The reaction mixture was stir at room temperature for 14 h and diluted with water. The aqueous layer was extracted with CH 2 CI 2 and the combined organic layers were washed with saturated ⁇ aHCO 3 and brine, dried over Na 2 SO 4 , and concentrated in vacuo to give a white solid. The white solid was suspended in H 2 SO 4 (cone.) (15 mL).
  • HNO 3 (cone.) (0.312 mL) was added to the suspension at 0°C.
  • the reaction mixture was slowly warmed to room temperature and stirred at room temperature for 4 h. After cooling to -10°C, the reaction mixture was neutralized with ammonium hydroxide and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over Na 2 SO 4 , and concentrated. The residue was purified by flash chromatography (1:9:5 MeOH / THF / hexane) to yield the title compound (0.61 g, 41%).
  • N-formyl-N, N', N'- trimethylethylenediamine (176 mL, 1.37 mol) dropwise at -78°C.
  • the reaction mixture was stirred for ca. 30 min at -78°C before warming to -23°C over ca. 30 min.
  • ethylene glycol dimethyl ether (1 L) followed by n-butyllifhium (2.5 M solution in hexane) (800 mL, 2.0 mol). The resulting mixture was stirred for ca. 2 h during which time the reaction mixture turned deep green.
  • a 12-L 4-necked round flask was charged with iodine (571 g, 2.25 mol) and ethylene glycol dimethyl ether (2 L) and the resultant solution was cooled to -78°C.
  • the contents of the 5-L flask were transferred via canula to the mixture of iodine and ethylene glycol dimethyl ether in the 12-L flask at -78°C.
  • the reaction mixture was stirred for an additional 1 h at -78°C.
  • the cooling bath was removed and the mixture was allowed to warm to about 0°C and treated with 2 L of water and 2 L of 1 N hydrochloric acid. Methyl t-butyl ether (2L) was added and the layers were separated.
  • 6-Imidazol-l-yl-2-(4-iodo-2-methoxy-pyridin-3-yl)-4-methyl-lH-benzimidazole To a solution of 5-imidazol-l-yl-3-methyl-benzene-l,2-diamine (175 mg, 0.93 mmol) in methanol (8 mL) was added a solution of 4-iodo-2-methoxy-pyridine-3-carbaldehyde (245 mg, 0.93 mmol) in methanol (5 mL) at 0°C. The reaction mixture was stirred at 0°C for 1.5 h and then at room temperature for 2 h.
  • N-(4-Cyano-2-methyl-6-nitro-phenyl)-2,2,2-trifluoro-acetamide To the ice-cold trifluoroacetic anhydride (60 mL) was added 4-amino-3-methyl-benzonitrile (14.33 g, 0.108 mol) in portion. The resulting white slurry was stirred at 0°C for 30 min. Then ammonium nitrate (17.28 g, 0.216 mol) was added. The reaction mixture was allowed to stir at 0°C for 1 h and at room temperature for 14 h. After removal of most solvent, the reaction mixture was cooled with ice and quenched with ice.
  • 6-Bromo-2-(4-iodo-2-methoxy-pyridin-3-yl)-4-methyl- lH-benzimidazole To a solution of 5-bromo-3-methyl-l,2-phenylenediamine (4 g, 19.9 mmol) in methanol (80 mL) was added 4-iodo-2-methoxy-pyridine-3-carbaldehyde (5.23 g, 19.9 mmol) in methanol (20 mL) dropwise at 0°C. The resulting slurry was stirred for 30 min at room temperature. Then iodine (2.53 g, 9.95 mmol) in methanol (20 mL) was added via a dropping funnel.
  • the reaction mixture was heated to reflux for 14 h under nitrogen and then cooled to room temperature. After removal of the solvent, the residue was diluted with ethyl acetate and washed with water. The aqueous fraction was extracted with ethyl acetate and the combined organic layers were washed with water and brine, dried over Na 2 SO 4 . After concentration in vacuo, the residue was purified by flash column chromatography (30% EtOAc / hexane) to yield the title compound (1 g, 73%) as a white solid.
  • the reaction mixture was heated to 80°C for 12 h, cooled to room temperature and concentrated with high vacuum.
  • the residue was purified by flash column chromatography (10% MeOH / CH 2 C1 2 ) to yield the title compound (0.90 g, 69%).
  • 2-(3-Amino-5-fluoro-2-nitro-phenyl)-malonic acid di-tert-butyl ester To the crude 2- (3,5-Difluoro-2-nitro-phenyl)-malonic acid di-tert-butyl ester (62g, 0.42 mol) was added 700 mL of 2M ammonia in methanol in a pressure bottle. The vessel was sealed and heated to 85°C for 18 hours. The reaction mixture was cooled and the vessel opened carefully and the methanol solution concentrated to provide 140 g of crude material.
  • 3-Amino-5-fluoro-2-nitro phenyl acetic acid To the 2-(3-Amino-5-fluoro-2-nitro- phenyl)-malonic acid di-tert-butyl ester (140g) in 500 mL of 4N HCl in dioxane was added 50 mL of water and heated to 40°C for 2 days. The solution was extracted with ethyl acetate (3X's) and the ethyl acetate washed with water (3X's) and brine.
  • 2-(4-Iodo-2-methoxy-pyridin-3-yl)-4-methyl-6-mo ⁇ holin-4-yl-lH-benzoimidazole 2-Methyl-4-mo ⁇ holin-4-yl-6-nitro-phenylamine (15.2 g, 64 mmol) was suspended in methanol (200 ml) in a PARR flask. Palladium on carbon (1.0 g, 10% Pd) was added and the suspension shaken under 60 psi of hydrogen overnight.
  • the mixture was filtered through a pad of celite (under argon) into a 3-neck flask, the celite rinsed with methanol and the filtrate diluted with methanol to a total volume of 500 ml and cooled to 0°C.
  • a solution of 4-Iodo-2-methoxy-pyridine-3-carbaldehyde (14.6 g, 55.5 mmol) in methanol (500 ml) was added slowly (during 3 hours). After addition of - X A of the solution the system was opened to air and stirred over the weekend, thereby reaching room temperature.
  • the resulting solution was then stirred at 0 °C for an additional 30 min,. the cooling bath was removed, and the reaction mixture was allowed to stir at room temperature in the presence of air for 72 h.
  • the resulting solution was concentrated in vacuo and the residue was dissolved in dichloromethane (1500 mL) and the solvent removed in vacuo (repeated 3x). The resulting dark foamy solid was dried under high vacuum.
  • the crude amine was dissolved in N,N-dimethylformamide (70 mL) and treated with the hydrochloride salt of ethyl mono-imido malonate (3.04 g, 15.48 mmol) and this mixture was stirred at 70°C for 2 hours.
  • the solvent was evaporated and the residue was dissolved in dichloromethane (100 mL) and treated with triethylamine (2.16 mL, 15.48 mmol), di-tert-butyl dicarbonate (3.38 g, 15.48 mmol) and dimethylaminopyridine (2 crystals).
  • the reaction was stirred at RT for 1.5 hours, then saturated ammonium chloride was added and the mixture was extracted with ethyl acetate (3x).
  • Examples 29-35 were prepared from commercially or readily available diamines which were prepared and condensed with 4-iodo-2-methoxy-pyridine-3-carbaldehyde as described in Scheme HI.
  • Examples 45 - 383 were prepared according to the general methods described above (Scheme HT)
  • LCMS conditions a) YMC C18 S5 4.6 x 50 mm; 0 - 100% gradient over 4 min*; 4 rnL/min flow rate b) YMC ODS-A C18 S7 3.0 x 50 mm; 0 - 100% gradient over 2 min*; 5 mL min flow rate c) YMC C18 S5 4.5 x 50 mm; 0 - 100% gradient over 8 min*; 2.5 mL/min flow rate d) YMC C18 S7 3.0 x 50 mm; 0 - 100% gradient over 3 min*; 5 mL/min flow rate e) YMC ODSA S3 6.0 xl50 mm; 0 - 100% gradient over 5 min*; 1.5 mL min flow rate f) PHS-PRJJVIESPHERE C184.6 x 30 mm; 0 - 100% gradient over 2 min*; 5 mL min flow rate g) YMC C18 S7 3.0 x 50 mm; 0
  • Example 520 (General procedure for examples 520-522)
  • the tiifluoroaceticacid salt of the pure title compound was dissolved in methanol and applied to a Narian Mega Bond-Elute SCX cartridge. Elution with methanol followed by 2.0 M ⁇ H3 / MeOH gave the free base. This material was suspended in MeOH and 1.00 N aqueous HCl (2 equiv.) was added.

Abstract

The present invention provides compounds of formula (I) and pharmaceutically acceptable salts thereof. The formula (I) compounds inhibit tyrosine kinase enzymes thereby making them useful as anti-cancer agents.

Description

NOVEL TYROSINE KINASE INHIBITORS
FIELD OF INVENTION The present invention relates generally to the field of tyrosine kinase enzyme inhibition using novel small molecules.
BACKGROUND OF THE INVENTION
Tyrosine Kinases are a class of enzymes, which catalyze the transfer of the terminal phosphate of adenosine triphosphate to the phenolic hydroxyl group of a tyrosine residue present in the target protein. Tyrosine kinases play a critical role in signal transduction for several cellular functions including cell proliferation, carcinogenesis, apoptosis, and cell differentiation (Plowman, G. D.; Ullrich, A.; Shawver, L. K.: Receptor Tyrosine Kinases As Targets For Drug Intervention. DNSP (1994) 7: 334-339). Therefore inhibitors of these enzymes would be useful for the treatment or prevention of proliferative diseases which are dependent on these enzymes. Strong epidemiologic evidence suggests that the overexpression or activation of receptor protein tyrosine kinases leading to constitutive mitogenic signaling is an important factor in a growing number of human malignancies. Tyrosine kinases that have been implicated in these processes include Abl, CDK's, EGF, EMT, FGF, FAK, Flk-1/KDR, HER-2, IGF-1R, IR, LCK, MET, PDGF, Src, and VEGF (Traxler, P.M. Protein Tyrosine Kinase Inhibitors in Cancer Treatment. Exp. Opin. Ther. Patents (1997) 7: 571-588; incorporated herein by reference). Hence, there is an ongoing need to investigate novel compounds that can be used to regulate or inhibit tyrosine kinase enzymes.
SUMMARY OF THE INVENTION
The present invention relates to compounds which inhibit tyrosine kinase enzymes, compositions which contain tyrosine kinase inhibiting compounds and methods of using inhibitors of tyrosine kinase enzymes to treat diseases which are characterized by an overexpression or upregulation of tyrosine kinase activity such as cancer, diabetes, restenosis, arteriosclerosis, psoriasis, angiogenic diseases and immunologic disorders (Powis, G.; Workman, P. Signaling targets For The Development of Cancer Drugs. Anti-Cancer Drug Design (1994), 9: 263-211; Merenmies, J.; Parada, L. E; Henkemeyer, M. Receptor Tyrosine Kinase Signaling in Vascular Development. Cell Growth Differ (1997) 8: 3-10; Shawver, L. K.; Lipsosn, K. E.; Fong, T. A. T.; McMahon, G.; Plowman, G. D.; Strawn, L. M. Receptor Tyrosine Kinases As Targets For Inhibition of Angiogenesis. Drug Discovery Today (1997) 2: 50-63; all herein incorporated by reference). A compound according to formula I
Figure imgf000003_0001
its enantiomers, diastereomers, pharmaceutically acceptable salts, hydrates, prodrugs and solvates thereof; wherein X is selected from the group consisting of N, C, Cι-C3 alkyl, Cι-C alkyl substituted with one or more R7, and a direct bond;
Y is selected from the group consisting of O and S ;
W is selected from the group consisting of N, C, O, and S; provided that if W is O or S, R9 is absent; R1, R2, R3, R4, R5, R6, R7, R , R9 are each independently selected from the group consisting of H, C1-6 alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, -OR60, -NO2, -OH, -SR60, -NR60R61 , -CN, -CO2R60, -CONR60R61, OCONR60R61, -NR62CONR60R61, -NR60SO2R61 , -SO2NR60R61, -SO2R63, - C(NR62)NR60R61, aryl, heteroaryl, -(CH2)nOR60, -(CH2)nNRo0R61, -(CH2)πSR60, - (CH2)n aryl, -(CH2)n heteroaryl, -(CH2)n heterocycloalkyl, -NH-Z-aryl, and -NH-Z- heteroaryl; wherein n is 1 to 3; and
Z is selected from the group consisting of - C4 alkyl, alkenyl, and alkynyl chain; Z having one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR60SO2R61 groups; Z optionally incorporating one or more groups selected from the group consisting of CO, CNOH, CNOR60, CNNR60 , CNNCOR60 and CNNSO2R60 ; and
R60, R61, R62, and R63 are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy, alkoxy, aryl, heteroaryl, heteroarylalkyl, and alkyl-R25;
R25 is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, - NR30COOR31, -NR30C(O)R31, -NR30SO2R31, -C(O)NR30R31, heteroaryl or heterocycloalkyl; and R30 and R31 are, independently, hydrogen, alkyl, or cycloalkyl.
In preferred embodiments, R1, R7, R8and R9 are H; R2 and R4 are H or F; Y is O; X is selected from the group consisting of N and CH; W is N; R5 is selected from the group consisting of H, methyl, ethyl, isopropyl, secondary butyl, cyclopropyl, F, and CF3; R6 is selected from the group consisting of H, 2-aminomethylpyridine,
NHCH2CH(OH)aryl, and NHCH(CH2OH)CH2aryl; and R3 is selected from the group consisting of OR60, C(NH)NHR60, C(O)NHR60 imidazole, imidazoline, tetrahydropyrimidine, piperazine, morpholine, homomorpholine, piperidine, pyrrolidine, homopiperazine and amino; wherein R60 is selected from the group consisting of H, alkyl, cycloalkyl, heterocycloalkyl, and alkyl-R25.
Preferably, R3 is -OR60 and R60 is alkyl, or -alkyl-R25, such as methyl, - (CH2)„CH2OH, or -(CH2)nCH2N(CH2CH2)2O, and n is 0, 1, or 2. In some embodiments, R25 is morpholine,
Figure imgf000004_0001
wherein R33, R34, R35, R36, and R37 are hydrogen, alkyl, or substituted alkyl. According to some embodiments of the present invention, R3 is piperazine, homopiperazine, 3-methylpiperazine, or 3,5-dimethylpiperazine being optionally substituted at the 4-N position with a compound selected from the group consisting of alkyl, cycloalkyl, cycloalkylalkyl, alkyl-R25, -C(O)-R15 , or -CO2R15 wherein R15 is hydrogen, alkyl, aryl, alkyl-R25, amino or aryl. Preferred piperazines are substituted with methyl, ethyl, CH2-cyclopropyl, hydroxyethyl, 2-dimethylaminoethyl, 2- diethylaminoethyl, 2-aminoethyl, 2-methylaminoethyl, 2-ethylaminoethyl, methoxyethyl, ethoxyethyl, thiomethoxyethyl, morpholine, and morpholinylethyl. R may be an amino group selected from the group consisting of hydroxyalkylamino, aminoalkylamino, dialkylaminoalkylamino, and heterocycloalkylalkylamino. Preferred amino groups include NHCH2CH2OH, NMeCH2CH2OH, NEtCH2CH2OH, NHCH2CH2NH2, NMeCH2CH2NH2, NEtCH2CH2NH2, NHCH2CH2NMe2, NMeCH2CH2NMe2, NEtCH2CH2NMe2, NHCH2CH2NEt2, NMeCH2CH2NEt2, NEtCH2CH2NEt2, NHCH2CH2N(CH2CH2)2O, NMeCH2CH2N(CH2CH2)2O, and NEtCH2CH2N(CH2CH2)2O.
According to some embodiments of the present invention, R3 is piperidine, optionally substituted with hydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, 1,3 dioxolane, 1,3 dioxane, -NHC(O)R15, -NHCO2R15, wherein R15 is hydrogen, alkyl, aryl or alkyl-R25 or morpholine, thiomorpholine, sulfoxymorpholine, sulfonylmorpholine, or homomorpholine, or a substituted morpholine, thiomorpholine, sulfoxymorpholine, sulfonylmorpholine, or homomorpholine. The morpholine, thiomorpholine, sulfoxymorpholine, sulfonyl morpholine, or homomorpholine is optionally substituted with hydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, alkyl-R25, -NHC(O)R15, -NHCO2R15, wherein R15 is hydrogen, alkyl, aryl or alkyl-R25.
In some embodiments of the present invention, R3 is an optionally substituted pyrrolidine that may be selected from the group consisting of 3-hydroxyl pyrrolidine, 3- alkoxy pyrrolidine, and 3-alkylamino pyrrolidine, 3-dialkylamino pyrrolidine.
According to one embodiment of the present invention, R6 is selected from the group consisting of H, 2-aminomethylpyridine, NHCH2CH(OH)aryl, and
NHCH(CH OH)CH2aryl. Preferred aryl groups include substituted phenyl wherein the phenyl is substituted with at least one Br, CI, F, -CN, methoxy, or-NHSO2CH3. According to some embodiments of the present invention, R is
Figure imgf000006_0001
wherein R17 is H, Br, CI, or F and R40 is H or alkyl.
Compounds of the present invention may have the formula:
Figure imgf000006_0002
In preferred embodiments, R12 is hydrogen, methyl, hydroxymethyl, methoxymethyl, CH2F, CH2CN, CO2H, or -CONR30R31; R13 is H or methyl; R17 is Br, F, or CI; R18 is methoxy or fluoro; and R19 is H; or R18 and R19 together form 4-O, 5 dihydrofuranyl.
According to one embodiment of the present invention, compounds have the formula:
Figure imgf000007_0001
wherein R51 is hydrogen, alkyl, aryl, or -alkyl-R25 and the remaining substituents are as defined above. In some embodiments, R51 is hydrogen, methyl, ethyl, or (CH2)nCH2-R25 wherein R25 is OH, OMe, F, CN, CF3, SOCH3 or SO2CH3, wherein n is 0 or 1. Preferably, R51 is methoxyethyl and R6 is -NHCH2CHOH-aryl.
According to some embodiments of the present invention, R51 is cyanoethyl, hydroxyethyl, CH2CH2SOCH3, CH2CH2CH2F, CH2CH2CH2CN, or CH2CH2CF3; R16 and R19are H; R17 is Br, or CI; and R18 is hydrogen or methoxy.
According to one embodiment of the present invention, compounds have the formula:
Figure imgf000007_0002
wherein
R'5 is hydrogen, alkyl, aryl or alkyl-R25 ; each R16 is independently hydrogen or methyl and the remaining substituents are as defined above. In preferred embodiments, R15 is hydrogen or methyl; R17 is bromo, chloro or fluoro; R18 is hydrogen or methoxy; and R19 is hydrogen .
According to one embodiment of the present invention, compounds have the formula:
Figure imgf000008_0001
wherein the substituents are as defined above. In preferred embodiments, R15 is hydrogen, methyl, ethyl, or -(CH2)nCH2-R25 wherein n is 0, 1, or 2; and R25 is OH, OMe, F, CN, CF3, SOCH3 or SO2CH3, -NR30COR31, -NR30COOR31, -NR30SO2R31, C(O)NR30R31, or has the formula:
Figure imgf000008_0002
wherein R33, R34, R35, R36, and R37 are hydrogen, alkyl, or substituted alkyl. In some preferred embodiments, R15 is ethyl, methoxyethyl, CH2CH2F, or CH2CH2CN; R17 is bromo or chloro; R18 is methoxy or hydrogen; and R19 is hydrogen.
According to some embodiments of the present invention, compounds have the formula:
Figure imgf000009_0001
wherein the substituents are as defined above. Preferably, R15 is -(CH2)nCH2R25 n is 0, 1, 2 or 3 and R25 is OH, OMe, F, CN, CF3, SOCH3 or SO2CH3 , -NR30COOR31, - NR30C(O)R31, -NR30SO2R31, -C(O)NR31R32 or has the formula:
Figure imgf000009_0002
wherein R33, R34, R35, R36, and R37 are hydrogen, alkyl, or substituted alkyl. More preferably, R15 is methyl, ethyl, CH2F, methoxyethyl, CH2CH2F, or
CH2CH2CH2SOCH3; R17 is bromo; R18 is hydrogen, methoxy, or fluoro; and R19 is H.
According to some embodiments of the present invention compounds have the formula:
Figure imgf000010_0001
In one preferred embodiment of the present invention, compounds have the following formula:
Figure imgf000010_0002
wherein
Y is selected from the group consisting of O and S; W is selected from the group consisting of N, C, O, and S; provided that if W is O or S, R9 is absent;
R1, R2, R3, R4, R5, R6, R8, R9 are each independently selected from the group consisting of H, C1- alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, -OR60, -NO2, -OH, -SR60, -NR60R61 , -CN, -CO2R60, -CONR60R61, OCONR60R61,
-NR62CONR60R61, -NR60SO2R61 , -SO2NR60R61, -SO2R63, -C(NR62)NR60R61, aryl, heteroaryl, -(CH2)nOR60, -(CH2)nNR60R61, -(CH2)nSR60, -(CH2)„ aryl, -(CH2)» heteroaryl, -(CH2)n heterocycloalkyl, -NH-Z-aryl, and -NH-Z-heteroaryl; wherein n is 1 to 3; and Z is selected from the group consisting of - C4 alkyl, alkenyl, and alkynyl chain; Z having one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR60SO2R61 groups; Z optionally incorporating one or more groups selected from the group consisting of CO, CNOH, CNOR60, CNNR60 , CNNCOR60 and CNNSO2R60 ; and R °, R61, R62, and R63 are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy, alkoxy, aryl, heteroaryl, heteroarylalkyl, and alkyl-R25;
R is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, - NR30COOR31, -NR30C(O)R31, -NR30SO2R31, -C(O)NR30R31, heteroaryl or heterocycloalkyl; and
R30 and R31 are, independently, hydrogen, alkyl, or cycloalkyl.
In addition to being used as single agents, it is contemplated that tyrosine kinase inhibitors can enhance the activity of cytotoxic or cytostatic treatments when used in combination with standard therapies known in the art. The invention also provides a pharmaceutical composition comprising a compound of formula I, as defined above, and a pharmaceutically acceptable carrier.
The invention further provides a pharmaceutical composition comprising a compound of formula I, as defined above, in combination with pharmaceutically acceptable carrier and at least one other anti-cancer agent optionally formulated as a fixed dose.
Additionally provided is a method of treating a condition associated with at least one tyrosine kinase enzyme comprising administering to a mammalian species in need of such treatment an effective amount of a compound of formula I, as defined above. Furthermore, the invention provides a method of treating a condition associated with at least one tyrosine kinase enzyme comprising administering to a mammalian species at least one other anti-cancer agent in combination with a compound of formula I, as defined above.
DESCRIPTION
The present invention provides for compounds of formula I, as defined above, pharmaceutical compositions employing such compounds and methods of using such compounds. Listed below are definitions of various terms used to describe the compounds of the instant invention. These definitions apply to the terms as they are used throughout the specification (unless they are otherwise limited in specific instances) either individually or as part of a larger group. The term "alkyl" herein alone or as part of another group refers to a monovalent alkane (hydrocarbon) derived radical containing from 1 to 12 carbon atoms unless otherwise defined. An alkyl group is an optionally substituted straight, branched or cyclic saturated hydrocarbon group. When substituted, alkyl groups may be substituted with up to four substituent groups, R as defined, at any available point of attachment. When the alkyl group is said to be substituted with an alkyl group, this is used interchangeably with "branched alkyl group". Exemplary unsubstituted such groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, and the like. Exemplary substituents may include but are not limited to one or more of the following groups: hydroxy, halo (such as F, CI, Br, I), haloalkyl (such as CCI3 or CF3), alkoxy, alkylthio, cyano, carboxy (-COOH), alkylcarbonyl (-C(O)R), alkoxycarbonyl (-OCOR), amino, carbamoyl (-NHCOOR or -OCONHR), urea (-NHCONHR), thiol, (-SH), sulfoxy, sulfonyl, aryl, heteroaryl, and heterocycloalkyl. Alkyl groups as defined may also comprise one or more carbon to carbon double bonds or one or more carbon to carbon triple bonds. Alkyl groups may also be represented by the formula alkyl-R25. In preferred embodiments, the alkyl group is a methyl, ethyl, propyl or butyl group and include substituted methyl, ethyl, propyl or butyl groups.
The term "alkenyl" herein alone or as part of another group refers to a hydrocarbon radical straight, branched or cyclic containing from 2 to 12 carbon atoms and at least one carbon to carbon double bond. An alkenyl group may be optionally substituted in the same manner as described for an alkyl group.
The term "alkynyl" herein alone or as part of another group refers to a hydrocarbon radical straight, branched or cyclic containing from 2 to 12 carbon atoms and at least one carbon to carbon triple bond. An alkynyl group may be optionally substituted in the same manner as described for an alkyl group. The term "alkoxy" as used alone or in combination herein refers to a straight or branched chain alkyl group covalently bonded to the parent molecule through an oxygen atom linkage containing from one to ten carbon atoms and the terms "C1-6 alkoxy" and "lower alkoxy" refer to such groups containing from one to six carbon atoms, examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy and the like. The term "optionally substituted" when used in connection with an alkoxy substituent refers to the replacement of up to two hydrogens, preferably on different carbon atoms with a radical selected form the group of lower alkyl, phenyl, cyano, halo, uϊfluoromethyl, nitro, hydroxy, alkanoyl, amino, monoalkyl amino and dialkylamino. Alkoxy groups may be substituted in the same manner that alkyl groups can be substituted as described above.
The term "sulfoxy" herein alone or as part of a group refers to -SO and may be substituted with, for example, alkyl or aryl groups.
The term "sulfonyl" herein alone or as part of a group refers to -SO2 and may be substituted with alkyl or aryl groups.
The term "amino" herein alone or as part of another group refers to -NH2. An "amino" may optionally be substituted with one or two substituents, which may be the same or different, such as alkyl, aryl, arylalkyl, alkenyl, alkynyl, heteroaryl, heteroarylalkyl, cycloheteroalkyl, cycloheteroalkylalkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, hydroxyalkyl, alkoxyalkyl or thioalkyl. Preferred substituents include alkylamino and dialkylamino, such as methylamino, ethyl amino, dimethylamino, and diethylamino. These substituents may be further substituted with a carboxylic acid or any of the alkyl or aryl substituents set out herein. In addition, the amino substituents may be taken together with the nitrogen atom to which they are attached to form 1- pyrrolidinyl, 1-piperidinyl, 1-azepinyl, 4-morpholinyl, 4-thiamorpholinyl, 4- sulfoxymorpholine, 4-sulfonylmorpholine, 1-piperazinyl, 4-alkyl-l-piperazinyl, 4- arylalkyl-1-piperazinyl, 4-diarylalkyl- 1-piperazinyl , 1-homopiperazinyl, 4-alkyl-l- homopiperazinyl, 4-arylalkyl- 1-homopiperazinyl, 4-diarylalkyl-l-homopiperazinyl; 1- pyrrolidinyl, 1-piperidinyl, or 1-azepinyl, optionally substituted with alkyl, alkoxy, alkylthio, halo, trifluoromethyl or hydroxy .
The term "aryl" herein alone or as part of another group refers to monocyclic or bicyclic aromatic rings, e.g. phenyl, substituted phenyl and the like, as well as groups which are fused, e.g., napthyl, phenanthrenyl and the like. An aryl group thus contains at least one ring having at least 6 atoms, with up to five such rings being present, containing up to 22 atoms therein, with alternating (resonating) double bonds between adjacent carbon atoms or suitable heteroatoms. Aryl groups may optionally be substituted with one or more groups including, but not limited to halogen, alkyl, alkenyl, alkynyl, alkoxy, hydroxy, carboxy, carbamoyl, alkyloxycarbonyl, alkylaminocarbonyl, nitro, trifluoromethyl, amino, cycloalkyl, cyano, alkyl S(O)m (m=O, 1, 2), or thiol. Aryl groups may also be substituted with heterocycloalkyl and heterocycloaryl groups to form fused rings, such as dihydiobenzfuranyl, oxindolyl, indolyl, indolinyl, oxindolyl, benzoxazolidinonyl, benzoxazolinyl and benzoxazolidinone.
The term "cycloalkyl" herein alone or as part of another group refers to fully saturated and partially unsaturated hydrocarbon rings of 3 to 9, preferably 3 to 7 carbon atoms. Further, a cycloalkyl may be substituted. A substituted cycloalkyl refers to such rings having one, two, or three substituents, preferably one, selected from the group consisting of halo, alkyl, substituted alkyl, alkenyl, alkynyl, nitro, cyano, oxo (=O), hydroxy, alkoxy, thioalkyl, -CO2H, -OC(=O)H, CO2-alkyl, - OC(=O)alkyl, =N-OH, =N-O-alkyl, aryl, heteroaryl, heterocyclo, a five or six membered ketal (i.e. 1,3-dioxolane or 1,3-dioxane), -NR'R", -C(=O)NR'R", - OC(=O)NR'R", -NR'CO2'R", -NR'C(=O)R", -SO2NR'R", and -NR'SO2'R", wherein each of R' and R" is independently selected from hydrogen, alkyl, substituted alkyl, and cycloalkyl, or R' and R" together form a heterocyclo or heteroaryl ring. Cycloalkyl groups may also be substituted with hetero atoms such as O, N, and S to form heterocycloalkyl groups. Preferred heterocycloalkyl groups include optionally substituted morpholine, homomorpholine (7 membered ring), thiomorpholine, piperazine, homopiperazine (7 membered ring), and piperidine.
The term "heteroaryl" herein alone or as part of another group refers to substituted and unsubstituted aromatic 5 or 6 membered monocyclic groups, 9 or 10 membered bicyclic groups, and 11 to 14 membered tricyclic groups which have at least one heteroatom (O, S or N) in at least one of the rings. Each ring of the heteroaryl group containing a heteroatom can contain one or two oxygen or sulfur atoms and/or from one to four nitrogen atoms provided that the total number of heteroatoms in each ring is four or less and each ring has at least one carbon atom. The fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or unsaturated. The nitrogen and sulfur atoms may optionally be oxidized and the nitrogen atoms may optionally be quatemized. Heteroaryl groups which are bicyclic or tricyclic must include at least one fully aromatic ring but the other fused ring or rings may be aromatic or non- aromatic. The heteroaryl group may be attached at any available nitrogen or carbon atom of any ring. The heteroaryl ring system may contain zero, one, two or three substituents selected from the group consisting of halo, alkyl, substituted alkyl, alkenyl, alkynyl, nitro, cyano, hydroxy, alkoxy, thioalkyl, -CO2H, -OC(=O)H, -CO2- alkyl, -OC(=O)alkyl, phenyl, benzyl, phenylethyl, phenyloxy, phenylthio, cycloalkyl, substituted cycloalkyl, heterocyclo, heteroaryl, -NR'R", -C(=O)NR'R", - OC(=O)NR'R", -NR'CO2'R", -NR'C(=O)R", -SO2NR'R", and -NR'SO2R", wherein each of R' and R" is independently selected from hydrogen, alkyl, substituted alkyl, and cycloalkyl, or R' and R" together form a heterocyclo or heteroaryl ring.
Exemplary monocyclic heteroaryl groups include pyrrolyl, pyrrolidinyl, imidazolinyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, and the like. Exemplary bicyclic heteroaryl groups include indolyl, indolinyl, oxindolyl, benzoxazolidinone, benzothiazolyl, benzodioxolyl, benzoxazolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, fiiropyridinyl, dihydroisoindolyl, tetrahydroquinolinyl and the like.
Exemplary tricyclic heteroaryl groups include carbazolyl, benzindolyl, phenanthrollinyl, acridinyl, phenanthridinyl, xanthenyl and the like.
The term "halogen" or "halo" herein alone or as part of another group refers to chlorine, bromine, fluorine or iodine selected on an independent basis. The term "hydroxy" herein alone or as part of another group refers to -OH.
The term "thioalkoxy" herein alone or as part of another group refers to an alkyl group as defined herein attached to the parent molecular group through a sulfur atom. Examples of thioalkoxy include, but are not limited to, thiomethoxy, thioethoxy, and the like.
Abbreviations: "Ph" represents phenyl; "Me" represents methyl; and "Et" represents ethyl.
An "anti-cancer agent" as used herein includes known anti-cancer treatments such as radiation therapy or with cytostatic or cytotoxic agents, such as for example, but not limited to, DNA interactive agents, such as cisplatin or doxorubicin; topoisomerase II inhibitors, such as etoposide; topoisomerase I inhibitors such as CPT-11 or topotecan; tubulin interacting agents, such as paclitaxel, docetaxel or the epothilones; hormonal agents, such as tamoxifen; thymidilate synthase inhibitors, such as 5-fluorouracil; anti-metabolites, such as methotrexate; tyrosine kinase inhibitors such as Iressa and OSI-774; angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDK inhibitors; Herl/2 inhibitors and monoclonal antibodies directed against growth factor receptors such as erbitux (EGF) and herceptin (Her2).
When a functional group is termed "protected", this means that the group is in modified form to preclude undesired side reactions at the protected site. Suitable protecting groups for the compounds of the present invention will be recognized from the present application taking into account the level of skill in the art, and with reference to standard textbooks, such as Greene, T. W. et al., Protective Groups in Organic Synthesis, Wiley, N.Y. (1991).
When C1-6 alkyl, alkenyl, alkynyl, cycloalkyl are substituted, they are preferably substituted with one or more hydroxy, cyano, carbamoyl, hydroxy, alkoxy, thiol, alkenyl, thioalkoxy, amino, alkylamino, amido, sulfonyl, sulfoxy, sulfonamido, halo, heterocycloalkyl, aryl or heteroaryl.
When aryl or heteroaryl are substituted, they are preferably substituted with one or more alkyl, alkenyl, alkynyl, cyano, carbamoyl, hydroxy, alkoxy, thioalkoxy, amino, amido, sulfonamido, halo or with R', R" wherein R', R" form a ring that is fused to the aryl group. When CH2aryl or heteroaryl are substituted, they are preferably substituted with one or more alkyl, alkyenyl, alkynyl, cyano, carbamoyl, hydroxy, alkoxy, thioalkoxy, amino, amido, sulfonamido, or halogen. When NH-Z-aryl or NH-Z-heteroaryl groups are substituted, they are preferably substituted with one or more alkyl, alkenyl, alkynyl, hydroxy, alkoxy, thioalkoxy, amino, halogen, nitro, nitrile, carboxylate, alkoxycarbonyl, carbamoyl, ester, amide, aryl, or heteroaryl The numbers in the subscript after the symbol "C" define the number of carbon atoms a particular group can contain. For example "C\_ alkyl" means a straight or branched saturated carbon chain having from one to six carbon atoms; examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, t- butyl, n-pentyl, sec-pentyl, isopentyl, and n-hexyl. Depending on the context, "Cj.g alkyl" can also refer to C . alkylene which bridges two groups; examples include propane- 1, 3 -diyl, butane- 1,4-diyl, 2-methyl-butane-l,4-diyl, etc. "C2-6 alkenyl" means a straight or branched carbon chain having at least one carbon-carbon double bond, and having from two to six carbon atoms; examples include ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, pentenyl, and hexenyl. Depending on the context, "C2-6 alkenyl" can also refer to C2-6 alkenediyl which bridges two groups; examples include ethylene-l,2-diyl (vinylene), 2-methyl-2-butene- 1,4-diyl, 2-hexene-l,6-diyl, etc. "C2-6 alkynyl" means a straight or branched carbon chain having at least one carbon-carbon triple bond, and from two to six carbon atoms; examples include ethynyl, propynyl, butynyl, and hexynyl. The term "alkyl-R25" includes optionally substituted alkyl groups such as methyl, ethyl, propyl, and butyl, attached to an R25group. R25generally includes hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, -NHCOOH, -NHC(O)-, -NHSO2-, -C(O)NH2, heteroaryl or heterocycloalkyl groups such as morpholinyl or group having the formula:
Figure imgf000017_0001
The terms "imidazole" and "imidazoline" herein alone or as part of another group includes substituted imidazoles and substituted imidazolines. Similarly, the term "tetrahydropyrimidine" includes substituted tetrahydropyrimidines. Likewise, the terms "piperazine", "piperidine" "morpholines", "homopiperazines", "homomoφholines" and "pyrrolidine" include substituted piperazines, substituted piperidines, substituted moφholines, substituted homomoφholines and substituted pyrrolidines, respectively.
Compounds of the present invention have the general formula I:
Figure imgf000018_0001
I its enantiomers, diastereomers, pharmaceutically acceptable salts, hydrates, prodrugs and solvates thereof;
wherein
X is selected from the group consisting of N, C, Cι-C3 alkyl, -C3 alkyl substituted with one or more R7, and a direct bond;
Y is selected from the group consisting of O and S ;
W is selected from the group consisting of N, C, O, and S, provided that when W is O or S, R9 is absent;
R1, R2, R3, R4, R5, R6, R7, R8, R9 are each independently selected from the group consisting of H, C1-6 alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, OR60, NO2, OH, SR60, NR60R61 , CN, CO2R60, CONR60R61, CO2NR60R61 , NR62CONR60R61, NR60SO2R61 , SO2NR60R61, C(NR62)NR60R61, aryl, heteroaryl, (CH2)nOR60, (CH2)nNR60R61 , (CH2)nSR60, (CH2)n aryl, (CH2)n heteroaryl, , (CH2)n heterocycloalkyl, NH-Z-aryl, and NH-Z-heteroaryl; wherein n is 1 to 3; and
Z is selected from the group consisting of - C4 alkyl, alkenyl, and alkynyl chain; Z having one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR60SO2R groups; Z optionally incoφorating one or more groups selected from the group consisting of CO, CNOH, CNOR60, CNNR60 , CNNCOR60 and CNNSO2R60 ; and R60, R61, R62 and R63 are independently selected from the group consisting of
H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy, alkoxy, aryl, heteroaryl, heteroarylalkyl, and alkyl-R wherein
R25 is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, - NR30COOR31, -NR30C(O)R31, -NR30SO2R31, -C(O)NR30R31, heteroaryl or heterocycloalkyl; and
R and R are, independently, hydrogen, alkyl, or cycloalkyl.
In some embodiments of the present invention, R3 is -OR60. R60 is alkyl, or - alkyl-R , wherein R is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, cyano, alkylsulfoxy, alkylsulfonyl, -R30COOR31, -NR30C(O)R31, - NR30SO2R31, -C(O)NR30R31, heteroaryl or heterocycloalkyl; and R30 and R3l are, independently, hydrogen, alkyl, or cycloalkyl. In preferred embodiments, R60 is methyl, -(CH2)nCH2OH, or -(CH2)nCH2N(CH2CH2)2O, and n is 0, 1, or 2.
In some embodiments, R is piperazine, homopiperazine, 3-methylpiperazine, or 3,5-dimethylpiperazine being optionally substituted at the 4-N position with a compound selected from the group consisting of alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, alkyl-R25, -C(O)-R15, or -CO2R15 wherein R15 is hydrogen, alkyl, aryl, alkyl-R25, amino or aryl; and R25 is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, cyano, halo, sulfoxy, sulfonyl, arylsulfonyl, -NR30COOR31, -NR30C(O)R31, -NR30SO2R31, -C(O)NR30R31, heteroaryl or heterocycloalkyl and R30 and R31 are, independently, hydrogen, alkyl, or cycloalkyl. In preferred embodiments, piperazine is substituted with Me, CH2cyclopropyl, CH2CH2NMe2, CH2CH2NEt2, CH2CH2NH2, CH2CH2NHMe, CH2CH2NHEt, N-CH2CH2N(CH2CH2)2O, (CH2)nCH2-R25 wherein R25 is OH, OMe, F, CN, CF3, SOCH3 or SO2CH3, wherein n is 0, 1 , or 2.
In some embodiments, R3 is an amino group. Preferred amino groups include NHCH2CH2OH, NMeCH2CH2OH, NEtCH2CH2OH, NHCH2CH2NH2, NMeCH2CH2NH2, NEtCH2CH2NH2, NHCH2CH2NMe2, NMeCH2CH2NMe2, NEtCH2CH2NMe2, NHCH2CH2NEt2, NMeCH2CH2NEt2, NEtCH2CH2NEt2, NHCH2CH2N(CH2CH2)2θ, NMeCH2CH2N(CH2CH2)2θ, NEtCH2CH2N(CH2CH2)2θ. In some embodiments, R3 is an optionally substitued piperidine. Preferred substituents are selected from the group consisting of hydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, 1,3 dioxolane (-OCHR15)2, 1,3 dioxane (-OCHR15CHR15CHR15O-) -NHC(O)R15, -NHCO2R15, wherein R15 is hydrogen, alkyl or alkyl-R25 .
In some embodiments R is an optionally substituted moφholine, homomoφholine, thiomoφholine, sulfoxymoφholine, or sulfonylmoφholine. Preferred substituents include hydroxy, thiol, amino, alkylamino, dialkylamino, alkoxy, thioalkoxy, alkyl-R25, -NHC(O)R15, -NHCO2R15, wherein R15 is hydrogen, alkyl or alkyl-R25 wherein R25.
In some embodiments, R3 is a pyrrolidine. Preferred pyrrolidines include, 3- hydroxyl pyrrolidine, 3- alkoxy pyrrolidine, and 3-alkylamino pyrrolidine.
According to one embodiment of the present invention, R3 is an optionally substituted N-tetrahydropyrimidine or N-imidazoline wherein the substituents are, preferably, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, cyanoalkyl, carboxyl, or carboxamide. In some embodiments, R6 is is selected from the group consisting of H, 2- aminomethylpyridine, NHCH2CH(OH)aryl, and NHCH(CH2OH)CH2aryl, wherein the aryl group is optionally substituted. In preferred embodiments, the aryl group is substituted with Br, CI, F, CN, or methoxy. In some embodiments, R6 has one of the following formulae:
Figure imgf000021_0001
wherein R40 is hydrogen or alkyl, preferably methyl, and R17 is hydrogen or halogen, such as Br, CI or F.
Preferred compounds of the present invention have one of the following formulae:
Figure imgf000022_0001
wherein
R12 and R13 are, independently, hydrogen, alkyl, or alkyl-R25;
R15is hydrogen, alkyl, or alkyl-R25;
R16 is independently, hydrogen or methyl;
R17, R18 and R19 are, independently, hydrogen, halogen, or alkoxy,or R18 and
R .19. together form a heterocycloalkyl or heteroaryl group; R25 is hydrogen, cycloalkyl, hydroxy, thiol, alkenyl, alkoxy, thioalkoxy, amino, halo, cyano, sulfoxy, sulfonyl, -NR30COOR31, -NR30C(O)R31, -NR30SO2R31,
C(O)NR )3j0υRr>31 , heteroaryl or heterocycloalkyl; and
R ,30 and R > 31 are, independently, hydrogen, alkyl, or cycloalkyl. In preferred embodiments, R12 is hydrogen, methyl, hydroxymethyl, methoxymethyl, alkyl-R25, CH2F, CH CN, CO2H, or -CONR30R31; R13 is H; R17 is Br, Cl or F;
1 R R is halo or methoxy; and
R19 is H.
According to one embodiment of the present invention, compounds have the formula:
Figure imgf000023_0001
wherein
Y is selected from the group consisting of O and S; W is selected from the group consisting of N, C, O, and S; provided that if W is O or S, R9 is absent;
R1, R2, R3, R4, R5, R6, R8, R9 are each independently selected from the group consisting of H, C1-6 alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, halo, amino, -OR60, -NO2, -OH, -SR60, -NR60R61 , -CN, -CO2R60, -CONR60R61, OCONR60R61,
-NR62CONR60R61, -NR60SO2R61 , -SO2NR60R61, -SO2R63, -C(NR62)NR60R61, aryl, heteroaryl, -(CH2)nOR60, -(CH2)nNR60R61, -(CH2)„SR60, -(CH2)n aryl, -(CH2)„ heteroaryl, -(CH2)n heterocycloalkyl, -NH-Z-aryl, and -NH-Z-heteroaryl; wherein n is 1 to 3; and Z is selected from the group consisting of C\ - C4 alkyl, alkenyl, and alkynyl chain; Z having one or more hydroxy, thiol, alkoxy, thioalkoxy, amino, halo, NR60SO2R61 groups; Z optionally incoφorating one or more groups selected from the group consisting of CO, CNOH, CNOR60, CNNR60 , CNNCOR60 and CNNSO2R60 ; and R60, R61, R62, and R63 are independently selected from the group consisting of H, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, hydroxy, alkoxy, aryl, heteroaryl, heteroarylalkyl, and alkyl-R25;
R is hydrogen, alkenyl, hydroxy, thiol, alkoxy, thioalkoxy, amino, alkylamino, dialkylamino, aryl, heteroaryl, cyano, halo, sulfoxy, sulfonyl, - NR30COOR31, -NR30C(O)R31, -NR30SO2R31, -C(O)NR30R31, heteroaryl or heterocycloalkyl; and
R and R are, independently, hydrogen, alkyl, or cycloalkyl. In preferred embodiments, R1 is -CO2R60, R2 , R4 ,R5 and R8 are H or methyl, R3 is a heterocycloalkyl and R6 is -NH(CH)2θH-aryl. Preferred heterocycles include moφholine or homomoφholine.
Suitable examples of salts of the compounds according to the invention include inorganic or organic acids. These include, but are not limited to, hydrochloride, hydrobromide, sulfate, methanesulfonate, maleate, fumarate, phosphate and other pharmaceutically acceptable salts. Salts which are unsuitable for pharmaceutical uses but which can be employed, for example, for the isolation or purification of free compounds of formula I or their pharmaceutically acceptable salts, are also included.
All stereoisomers of the compounds of the instant invention are contemplated, either in admixture or in pure or substantially pure form. The definition of the compounds according to the invention embraces all possible stereoisomers and their mixtures. It very particularly embraces the racemic forms and the isolated optical isomers having the specified activity. The racemic forms can be resolved by physical methods, such as, for example, fractional crystallization, separation or crystallization of diastereomeric derivatives or separation by chiral column chromatography. The individual optical isomers can be obtained from the racemates by conventional methods, such as, for example, salt formation with an optically active acid followed by crystallization.
It should be understood that the present invention includes prodrug forms of the compounds of formula I. Various forms of prodrugs are well known in the art. For examples of such prodrug derivatives, see: (a) Design of Prodrugs, edited by H. Bundgaard (Elsevier, 1985); and Methods in Enzymology, Vol. 42, pp. 309-396, edited by K.
Widder et al., (Academic Press, 1985);
(b) A Textbook of Drug Design and Development, edited by Krosgaard-Larsen and H. Bundgaard, Chapter 5, "Design and
Application of Prodrugs," by H. Bundgaard, pp. 113-191 (1991);
(c) H. Bundgaard, Advanced Drug Deliver Reviews, 8, pp. 1-38 (1992);
(d) H. Bundgaard et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); and
(e) N. Kayeka et al., Chem. Phar. Bull., 32, 692 (1984). The invention also provides a pharmaceutical composition comprising a compound of formula I, as defined above, and a pharmaceutically acceptable carrier and at least one other anti-cancer agent formulated as a fixed dose. Preferred anti- cancer agents are selected from the group consisting of: tamoxifen, toremifen, raloxifene, droloxifene, iodoxyfene, megestrol acetate, anastrozole, letrazole, borazole, exemestane, flutamide, nilutamide, bicalutamide, cyproterone acetate, goserelin acetate, luprolide, finasteride, herceptin, methotrexate, 5-fluorouracil, cytosine arabinoside, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin, cisplatin, carboplatin, melphalan, chlorambucil, busulphan, cyclophosphamide, ifosfamide, nitrosoureas, thiotephan, vincristine, taxol, taxotere, etoposide, teniposide, amsacrine, irinotecan, topotecan, an epothilone; a tyrosine kinase inhibitor such as Iressa or OSI-774; an angiogenesis inhibitor; an EGF inhibitor; a VEGF inhibitor; a CDK inhibitor; a Herl/2 inhibitor and monoclonal antibodies directed against growth factor receptors such as erbitux (EGF) and herceptin (Her2). The invention further provides a method of treating a condition via modulation of at least one tyrosine kinase enzyme comprising administering to a mammalian species in need of such treatment an effective amount of a compound of formula I, as defined above.
Additionally, the invention provides a method of treating a condition via modulation of at least one tyrosine kinase enzyme comprising administering to a mammalian species in need of such treatment an effective amount of a compound of formula I, as defined above, in combination (simultaneously or sequentially) with at least one other anti-cancer agent.
A preferred condition, treated by said methods of the instant invention, is cancer. Additionally, the tyrosine kinase enzyme may include (but is not limited to): Abl, CDK's, EGF, EMT, FGF, FAK, Flk-1/KDR, HER-2, IGF-1R, IR, LCK, MET, PDGF, Src, and VEGF.
The invention also provides a method for treating cancer, comprising administering to a mammalian species in need of such treatment, a therapeutically effective amount of at least one of the pharmaceutical compositions defined above. The invention further provides a method for treating proliferative diseases, comprising administering to a mammalian species in need of such treatment a therapeutically effective amount of at least one of the pharmaceutical compositions defined above. Certain compounds of formula I may generally be prepared according to the following schemes and the knowledge of one skilled in the art. Solvates (e.g., hydrates) of the compounds of formula I are also within the scope of the present invention. Methods of solvation are generally known in the art. Accordingly, the compounds of the instant invention may be in the free or hydrate form, and may be obtained by methods exemplified by the following schemes below.
More specifically, Schemes I-Vπ illustrate the preparation of compounds claimed in this invention. The examples, which follow, illustrate the compounds that can be synthesized by these schemes. The schemes are not limited by the examples listed or by any substituents employed for illustrative puφoses. Scheme I describes the preparation of the benzimidazoles. The starting diamines 1 are readily available using literature methods or are obtained commercially. The diamine is then condensed with an aldehyde 2 to provide the benzimidazole 3. Further modification of the functional groups on the aryl group of the benzimidazole or heterocycle of 3 are then envisioned.
Scheme I.
Figure imgf000027_0001
Alternatively, the benzimidazole could be formed in a step-wise manner (see Scheme H) by amide formation using the acid chloride of 5 or any of the commonly used peptide coupling reagents such as DCC (dicyclohexylcarbodiimide), EDCI (l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), etc. Once the amide 6 was formed the nitro group could be reduced using catalytic hydrogenation, transfer hydrogenation or chemical reduction such as SnC^ or iron powder or other methods known in the art for reduction of aryl nitro groups. Treatment of the aniline with acid would then form the benzimidazole.
Scheme II.
Figure imgf000027_0002
For example, Scheme HI illustrates the use of 4-iodo-2-methoxy-pyridine-3- carbaldehyde 7 to provide the functionalized benzimidazole 8. Hydrolysis of the methoxy group using protic acid conditions, TMSI (trimethylsilyl iodide), BBr3, or other conditions known in the art for cleaving a methyl ether would provide the halopyridone 9. Addition of heteroatom nucleophiles using amines, alcohols or thiols would then provide the substituted pyridones 10. Other functionality could be incoφorated into the aldehyde and the above example is included for illustrative puφoses only.
Scheme III
Figure imgf000029_0001
9 10
Likewise the aryl ring of the benzimidazole prepared using Schemes I or II can be modified. For example introduction of a cyano group for R3 on the benzimidazole allows for the formation of heterocycles such as imidazole, imidazolines, oxazolines, thiazolines, amides, or amidines. Scheme IV illustrates such transformations. Starting from the cyano-substituted benzimidazole 11 the heterocycle can be modified as illustrated in Scheme IV to provide 12. Imidate formation preferably using ethanol and acid provides intermediate 13. Imidate 13 can be transformed using diamines to form imidazolines, amino alcohols to form oxazolines, amino acetals to form imidazoles, and amino thiols to form thiazolines 14. Alternatively the imidate can be hydrolyzed to the acid and coupled with amines using any of the standard amide formation reagents (DCC, EDCI, etc.) to form amides 15. Imidate 13 is also a useful intermediate for the preparation of amidines 16 by reacting with amines.
Scheme IV
Figure imgf000030_0001
15 X= NH, O, S
Scheme V illustrates further transformation of benzimidazoles that bear a halogen atom using palladium catalysis using conditions developed by Suzuki [Yang et al. Acta Chem. Scand. (1993) 221; Suzuki et al. Synth. Commun. (1981) 11: 513] or Buchwald/Hartwig [Buchwald et al. J. Am. Chem. Soc. (1994) 116: 7901; Hartwig et al. J. Am. Chem. Soc. (1994) 116: 5969; Hartwig. Angew. Chem., Int. Ed. Engl. (1998) 37: 2046] and variations of these methods. Preparation of a bromide substituted benzimidazole 17 could be envisioned to provide a substrate for Suzuki coupling with aryl, vinyl, and heterocyclic boronic acids to provide benzimidazoles 18. Likewise, amines and heterocycles such as piperazine or moφholine derivatives 19 can be prepared from the same bromide using amines under conditions described by Buchwald and Hartwig or variations thereof. Scheme V
Figure imgf000031_0001
17 HNYZ, Pd
Figure imgf000031_0002
Figure imgf000031_0003
18 19
X = NH, 0, S
Alternatively amine and heterocyclic derivatives such as 19 can be prepared using intermediate 6 described in Scheme II. When the R3 of 6 is a halogen, preferably F, the halogen can be displaced with amines, alcohols, heterocyclic amines and other nitrogen containing heterocycles such as piperazine, piperidine, 4-amino piperidine, moφholine, imidazole, etc (Scheme VI). The terminal nitrogen of piperazine or 4-amino piperidine can then be alkylated using standard alkylation conditions or reacted with aldehydes in a reductive amination reaction to provide alkylated derivatives. Alternatively the terminal nitrogen atom of piperazine or 4- amino piperidine can be acylated or carbamoylated using any number of conditions that are routine for someone skilled in the art of organic synthesis. Following the example illustrated in Scheme JJ compounds such as 19 could be prepared.
Scheme VI.
Figure imgf000031_0004
19 Alternatively amines, heterocycles, and alcohols can be introduced at R3 using a nucleophilic aromatic substitution reaction started from an intermediate 20 were R3 is halogen, preferably F, the halogen can be displaced with amines, alcohols, heterocyclic amines and other nitrogen containing heterocycles such as piperazine, piperidine, 4-amino piperidine, moφholine, imidazole, etc (Scheme VH). The terminal nitrogen of piperazine or 4-amino piperidine can then be alkylated using standard alkylation conditions or reacted with aldehydes in a reductive animation reaction to provide alkylated derivatives. Alternatively the terminal nitrogen atom of piperazine or 4-amino piperidine can be acylated or carbamoylated using any number of conditions that are routine for someone skilled in the art of organic synthesis. The resulting nitro aniline could be reduced to the diamine 21 and processed as illustrated in Scheme DI.
Scheme VH
Figure imgf000032_0001
21
20
Alternatively, a completely different approach can be envisioned for the synthesis of some of the compounds of Formula I as illustrated in Scheme VIE. The same diamines 1 can be condensed with imidate 22 to give esters 23. Acylation of compound 23 (R10 = H) with various amino-esters of type 24 under acylating conditions known in the art may afford compounds 25. Cyclization may then be performed using a base such as sodium hydride, potassium tert-butoxide, lithium bis(trimethylsilyl)amide or the like to give compounds 26 which will then be converted to the corresponding sulfonates 27 by reaction with various alkylsulfonyl halides or alkylsulfonic anhydrides such as methanesulfonyl chloride, p- toluenesulfonyl chloride or preferably trifluoromethanesulfonic anhydride in presence of a weak base such as pyridine, 2,6-di-tert-butyl-4-methyl-pyridine or triethylamine. Substitution with heteroatom nucleophiles such as amines, thiols or alcohols provides the desired 5-, 6- or 7-membered cyclic cores 28.
Scheme VHI
Figure imgf000033_0001
Utility
The compounds according to the invention have pharmacological properties; in particular, the compounds of formula I are tyrosine kinase enzyme inhibitors. The novel compounds of formula I are thus useful in the therapy of a variety of proliferative diseases (including but not limited to diseases associated with tyrosine kinase enzymes) such as cancer, autoimmune diseases, viral diseases, fungal diseases, neurodegenerative disorders and cardiovascular disease.
More specifically, the compounds of formula I are useful in the treatment of a variety of cancers, including, but not limited to, the following: a) carcinoma, including that of the bladder, breast, colon, kidney, liver, lung, including small cell lung cancer, esophagus, gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, including squamous cell carcinoma; b) hematopoietic tumors of lymphoid lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkins lymphoma, non-Hodgkins lymphoma, hairy cell lymphoma and Burkett's lymphoma; c) hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias, myelodysplastic syndrome and promyelocytic leukemia; d) tumors of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; e) tumors of the central and peripheral nervous system, including astrocytoma, neuroblastoma, glioma and schwannomas; and f) other tumors, including sacroma, melanoma, seminoma, teratocarcinoma, osteosarcoma, xenoderoma pigmentosum, keratoctanthoma, thyroid follicular cancer and Kaposi's sarcoma.
Due to the key role of tyrosine kinases in the regulation of cellular proliferation in general, inhibitors could act as reversible cytostatic agents which may be useful in the treatment of any disease process which features abnormal cellular proliferation, e.g., benign prostatic hypeφlasia, familial adenomatosis polyposis, neuro-fibromatosis, atherosclerosis, pulmonary fϊbrosis, arthritis, psoriasis, glomerulonephritis, restenosis following angioplasty or vascular surgery, hypertrophic scar formation, inflammatory bowel disease, transplantation rejection, endotoxic shock, and fungal infections. Compounds of formula I may induce apoptosis. The apoptotic response is aberrant in a variety of human diseases. Compounds of formula I, as modulators of apoptosis, will be useful in the treatment of cancer (including but not limited to those types mentioned herein above), viral infections (including but not limited to heφesvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus), prevention of AIDS development in HlV-infected individuals, autoimmune diseases (including but not limited to systemic lupus, erythematosus, autoimmune mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease, and autoimmune diabetes mellitus), neurodegenerative disorders (including but not limited to Alzheimer's disease, AIDS -related dementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentosa, spinal muscular atrophy and cerebellar degeneration), myelodysplastic syndromes, aplastic anemia, ischemic injury associated with myocardial infarctions, stroke and reperfusion injury, arrhythmia, atherosclerosis, toxin-induced or alcohol related liver diseases, hematological diseases (including but not limited to chronic anemia and aplastic anemia), degenerative diseases of the musculoskeletal system (including but not limited to osteoporosis and arthritis) aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple sclerosis, kidney diseases and cancer pain.
Compounds of formula I may modulate the level of cellular RNA and DNA synthesis. These agents would therefore be useful in the treatment of viral infections (including but not limited to HIV, human papilloma virus, heφesvirus, poxvirus, Epstein-Barr virus, Sindbis virus and adenovirus).
Compounds of formula I may also be useful in the chemoprevention of cancer. Chemoprevention is defined as inhibiting the development of invasive cancer by either blocking the initiating mutagenic event or by blocking the progression of pre- malignant cells that have already suffered an insult or inhibiting tumor relapse.
Compounds of formula I may also be useful in inhibiting tumor angiogenesis and metastasis.
The compounds of this invention may also be useful in combination (administered together or sequentially) with known anti-cancer treatments such as radiation therapy or with cytostatic or cytotoxic agents, such as for example, but not limited to, DNA interactive agents, such as cisplatin or doxorubicin; topoisomerase II inhibitors, such as etoposide; topoisomerase I inhibitors such as CPT-11 or topotecan; tubulin interacting agents, such as paclitaxel, docetaxel or the epothilones; hormonal agents, such as tamoxifen; thymidilate synthase inhibitors, such as 5-fluorouracil; and anti-metabolites, such as methotrexate; tyrosine kinase inhibitors such as Iressa and OSI-774 (Tarceva1"1) ; Avastin, Herceptin, angiogenesis inhibitors; EGF inhibitors; VEGF inhibitors; CDK inhibitors; Herl/2 inhibitors and monoclonal antibodies directed against growth factor receptors such as erbitux (EGF) and herceptin (Her2).
If formulated as a fixed dose, such combination products employ the compounds of this invention within the dosage range described below and the other pharmaceutically active agent or treatment within its approved dosage range. Compounds of formula I may also be administered sequentially with known anticancer or cytotoxic agents when a combination formulation is inappropriate. The invention is not limited in the sequence of administration; compounds of formula I may be administered either prior to or after administration of the known anticancer or cytotoxic agent(s).
Further subject matter of the invention also includes pharmaceuticals for use, as described above, including controlling cancer, inflammation and arthritis, which contain at least one compound of the formula I as defined above or at least one of its pharmacologically acceptable acid addition salts, and the use of a compound of the formula I as defined above for the preparation of a pharmaceutical having activity against proliferative diseases as described previously including against cancer, inflammation and/or arthritis.
The pharmacological properties of the compounds of this invention may be confirmed by a number of pharmacological assays. The exemplified pharmacological assays which follow have been carried out with the compounds according to the invention and their salts.
Biological Assays A. CDK 2/cyclin E Kinase Assay
Kinase reactions consisted of 5 ng of baculovirus expressed GST- CDK2/cyclin E complex, 0.5 μg GST-RB fusion protein (amino acids 776-928 of retinoblastoma protein), 0.2 μCi 33P γ-ATP, 25 μM ATP in 50 μl kinase buffer (50 mM Hepes, pH 8.0, 10 mM MgCl2) 1 mM EGTA, 2 mM DTT). Reactions were incubated for 45 minutes at 30° C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration of 15%. TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT). Dose response curves were generated to determine the concentration required inhibiting 50% of kinase activity (IC50). Compounds were dissolved at 10 mM in dimethylsulfoxide (DMSO) and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equaled 2%. IC50 values were derived by non-linear regression analysis and have a coefficient of variance (SD/mean, n=6) = 14%.
B. EMT Kinase Assay
A filter-based kinase assay, measuring the phosphorylation of Gst-SLP76 by Gst-Emtk, was employed to determine the compound inhibitory activity against Emt. The kinase reaction was performed in a 96-well plate at room temperature for 15 min before being terminated by adding 100 μl of 20% trichloroacetic acid (TCA) containing 0.1 M sodium pyrophosphate. The kinase reaction mixture (60 μl ) contained 25 mM HEPES, pH 7.0, 0.1 mg/ml BSA, 5 mM MgCl2, 5 mM MnCl2, 8 ng of enzyme (Gst-Emtk), 5 μg of the substrate protein (Gst-SLP76), 1 μM ATP, 0.4 μCi of [γ-P33] ATP and the tested compound (at various concentrations). After termination, the proteins were allowed to precipitate in the presence of TCA for 1 hr at 4 °C. The precipitated proteins were then harvested on a filter plate (UniFilter-96, GF/C, Packard Instrument) and washed to remove excess [γ-P33]ATP. The radioactivity was determined using a TopCount NXT (Packard Instrument) after adding 35 μl of Microscint 20 (Packard instrument).
C. FAK Tyrosine Kinase Assay
The Focal Adhesion kinase was assayed using the synthetic polymer poly(Glu/Tyr) (Sigma Chemicals) as a phosphoacceptor substrate. Each reaction mixture consisted of a total volume of 50 ul and contained 100 ng of baculovirus- expressed enzyme, 2μg of ρoly(Glu/Tyr), lμM of ATP, and 0.2 μCi of [γ-33P]ATP. The mixtures also contained 40 mM Tris.HCl, pH 7.4, 1 mM MnCl2, 0.5 mM DTT, and 0.1 mg/ml bovine serum albumin. The reaction mixtures were incubated at 26°C for 1 hour and kinase activity was determined by quantitation of the amount of radioactive phosphate transferred to the poly(Glu/Tyr) substrate. Incoφoration was measured by the addition of cold trichloroacetic acid (TCA) precipitation of the proteins which were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT). Compounds were dissolved in dimethyl sulfoxide to a concentration of 10 mM and were evaluated at six concentrations, each in triplicate. The final concentration of DMSO added to the kinase assays was 0.5%, which has been shown to have no effect on kinase activity. IC50 values were derived non-linear regression analysis and have a coefficient of variance (SD/mean, n=6) = 16%.
D. HER-l/HER-2 Kinase Assay
Kinase reactions consisted of 10 ng of baculovirus expressed GST- HER1, 100 ng of HER2, 100 ng/ml poly(Glu/Tyr) (Sigma), 0.2 μCi 33P γ-ATP, 1 μM ATP in 50 μl kinase buffer (50 mM Tris, pH 7.5, 10 mM MnC12, 0.5 mM DTT). Reactions were incubated for lh at 27 C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration 15%. TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co.,
Meriden, CT). Dose response curves were generated to determine the concentration required to inhibit 50% of kinase activity (IC50). Compounds were dissolved at 10 mM in dimethylsulfoxide (DMSO) and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equaled 1%. IC5Q values were derived by non-linear regression analysis and have a coefficient of variance (SD/mean, n=6) = 16%.
E. IGF- Receptor Tyrosine Kinase Assay The IGF-1 receptor tyrosine kinase was assayed using the synthetic polymer poly(Glu Tyr) (Sigma Chemicals) as a phosphoacceptor substrate. Each reaction mixture consisted of a total volume of 50 ul and contained 125 ng of baculovirus expressed enzyme, 2.5μg of poly(Glu/Tyr), 25μM of ATP, and 0.1 μCi of [γ- 33P]ATP. The mixtures also contained 20 mM MOPS, pH 7.0, 5 mM MnCl2, 0.5 mM DDT, and 0.1 mg/ml bovine serum albumin. The reaction mixtures were incubated at
30 C for 45 minutes and kinase activity was determined by quantitation of the amount of radioactive phosphate transferred to the poly(Glu/Tyr) substrate. Incoφoration was measured by the addition of cold trichloroacetic acid (TCA) precipitation of the proteins which were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT). Compounds were dissolved in dimethyl sulfoxide to a concentration of 10 mM and were evaluated at six concentrations, each in triplicate. The final concentration of DMSO added to the kinase assays was 0.5%, which has been shown to have no effect on kinase activity. IC50 values were derived by non-linear regression analysis and have a coefficient of variance (SD/mean, n=6) = 16%.
F. Insulin Receptor Tyrosine Kinase Assay
The Insulin Receptor Tryrosine kinase was assayed using the synthetic polymer poly(Glu/Tyr) (Sigma Chemicals) as a phosphoacceptor substrate. Each reaction mixture consisted of a total volume of 50 ul and contained 90 ng of baculovirus-expressed enzyme, 2.5μg of poly(Glu/Tyr), 25μM of ATP, and 0.1 μCi of [γ-33P]ATP. The mixtures contained also 20 mM Tris.HCl, pH 7.4, 5 mM MnCl2, 0.5 mM DTT, and 0.1 mg/ml bovine serum. The reaction mixtures were incubated at 26°C for 1 hour and kinase activity was determined by quantitation of the amount of radioactive phosphate transferred to the poly(Glu/Tyr) substrate. Incoφoration was measured by the addition of cold trichloroacetic acid (TCA) precipitation of the proteins which were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT). Compounds were dissolved in dimethyl sulfoxide to a concentration of 10 mM and were evaluated at six concentrations, each in triplicate. The final concentration of DMSO added to the kinase assays was 0.5%, which has been shown to have no effect on kinase activity. IC50 values were derived non-linear regression analysis and have a coefficient of variance (SD/mean, n=6) = 16%.
G. LCK Kinase Assay
Kinase reactions consisted of 10 ng of baculovirus expressed lOng GST-Lck, 100 ng/ml poly(Glu/Tyr) (Sigma), 0.2 μCi 33P γ-ATP, 1 μM ATP in 50 μl kinase buffer (50 mM Tris, pH 7.5, 10 mM MnC12, 0.5 mM DTT). Reactions were incubated for lh at 27 C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration 15%. TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co.,
Meriden, CT). Dose response curves were generated to determine the concentration required to inhibit 50% of kinase activity (IC5Q). Compounds were dissolved at 10 mM in dimethyl sulfoxide (DMSO) and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equaled 1%. IC5o values were derived by non-linear regression analysis and have a coefficient of variance (SD/mean, n=6) = 16%.
H. MET Kinase Assay Kinase reactions consisted of lOng of baculovirus expressed GST-Met, 2.5ug poly(Glu/Tyr) (Sigma), 0.2μCi 33P γ-ATP, 10 μM ATP in 50 μl kinase buffer (40mM Tris, pH 7.5, ImM MnC12, 0.50 mM DTT). Reactions were incubated for lh at 27 C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration 3.5%. TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co., Meriden, CT). Dose response curves were generated to determine the concentration required to inhibit 50% of kinase activity (IC50). Compounds were dissolved at 10 mM in dimethylsulfoxide (DMSO) and evaluated at seven concentrations, each in triplicate.
The final concentration of DMSO in the assay equaled 1%. IC50 values were derived by non-linear regression analysis and have a coefficient of variance (SD/mean, n=6) =
16%.
I. PDGF Receptor Kinase Assay
Kinase reactions consisted of 70ng of baculovirus expressed GST-PDGFbR,
0.3ug biotinylated poly(Glu/Tyr) (Sigma), in 50 μl kinase buffer (20 mM Hepes, pH 7.5, 0.7uM ATP, lOmM MnC12, 0.5mM DTT, 0.15mM NaCl, O.lmg/ml BSA).
Reactions were incubated for 30 minutes at room temperature with shaking and stopped by the addition of lOul of 0.2M EDTA, ρH8.0. 150ul of HTRF detection buffer was added and incubated for 1 hour and 30 minutes at room temperature.
Counts were quantitated on Discovery HTRF Packard Instrument.
J. VEGFR-2 (KDR) Kinase Assay
Kinase reactions consisted of 7.5ng of baculovirus expressed GST-KDR, 1.5ug poly(Glu/Tyr) (Sigma), 0.04μCi 33P γ-ATP, 2.5 μM ATP in 50 μl kinase buffer (25 mM Tris, pH 7.5, 1.8 mM MnC12, 0.0.625 mM DTT). Reactions were incubated for lh at 27 C and stopped by the addition of cold trichloroacetic acid (TCA) to a final concentration 15%. TCA precipitates were collected onto GF/C unifilter plates (Packard Instrument Co., Meriden, CT) using a Filtermate universal harvester (Packard Instrument Co., Meriden, CT) and the filters were quantitated using a TopCount 96-well liquid scintillation counter (Packard Instrument Co.,
Meriden, CT). Dose response curves were generated to determine the concentration required to inhibit 50% of kinase activity (IC50). Compounds were dissolved at 10 mM in dimethylsulfoxide (DMSO) and evaluated at six concentrations, each in triplicate. The final concentration of DMSO in the assay equaled 1%. IC50 values were derived by non-linear regression analysis and have a coefficient of variance (SD/mean, n=6) = 16%. K. Cytotoxicity assay (HT-29-colon; Colo205, MCF-7-breast)
Tumor cell lines are maintained in McCoy's 5 A medium (GIBCO) and 10% heat inactivated fetal bovine serum (GIBCO). The in vitro cytotoxicity is assessed in tumor cells by a tetrazolium-based colorimetric assay which takes advantage of the metabolic conversion of MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulphenyl)-2H-tetrazolium, inner salt) (Promega) to a reduced form that absorbs light at 492 nm (1). Cells are seeded 24 hr prior to drug addition. Following a 72 hour incubation at 37°C with serially diluted test compound, MTS (Riss, T.L, et al., Comparison of MTT, XTT, and a novel tetrazolium compound MTS for in vitro proliferation and chemosensitivity assays.," Mol. Biol. Cell 3 (Suppl.):184a, 1992), in combination with the electron coupling agent phenazine methosulfate, is added to the cells. The incubation is continued for 3 hours, then the absorbency of the medium at 492 nm is measured with a spectrophotometer to obtain the number of surviving cells relative to control populations. The results are expressed as median cytotoxic concentrations (IC50 values).
Biological Activity (uM); compounds of the present invention had kinase activity of <25uM against one or more of the following kinases CDK, EMT, FAK, Herl, Her2, IGF, IR, LCK, MEK, MET, PDGF, VEGF. HT-29 and Colo205 are human colon tumor cell lines, and MCF-7 is a human breast tumor cell line.
General Procedure for the Preparation of 2-Hydroxy-2-(substituted-phenyl)- ethylamines:
Figure imgf000042_0001
l-(3-Chloro-phenyl)-2-nitro-ethanol: To a solution of 3-chloro-benzaldehyde (20 g, 0.142 mol) in nitromethane (100 mL) were added magnesium sulfate (37.6 g, 0.312 mol) and phosphazene base Prt-bu-tris etramethylene) (4.43 g, 0.014 mol). The reaction mixture was stirred at room temperature for 2 h. After concentration in vacuo, the residue was purified by flash chromatography (25% EtOAc / hexane) to yield the title compound (26.4 g, 100%) as a green-yellow oil. 1H NMR (300 MHz, DMSO-d6) 57.53 (IH, s), 7.35 - 7.42 (3H, m), 6.23 (IH, broad s), 5.32 - 5.33 (IH, m), 4.90 (IH, dd, /= 3.2, 12.4 Hz), 4.60 (IH, dd, J = 1.2, 12.4 Hz).
Figure imgf000043_0001
[l-(3-Chloro-phenyl)-2-nitro-ethoxy]-triethyl-silane: To a solution of l-(3-chloro- phenyl)-2-nitro-ethanol (26 g, 0.14 mol) in DMF (50 mL) were added imidazole (28.6 g, 0.42 mol) and chlorotriethylsilane (25.3 g, 0.17 mol). The reaction mixture was stirred at room temperature for 2 h. After quenching with water, the mixture was extracted with ethyl acetate. The combined organic layers were washed with water and brine, dried over Na2SO4, and filtered. After removal of solvent, the crude product was purified by flash chromatography (2% EtOAc / hexane) to yield the title compound (37 g, 91%) as a colorless oil. *H NMR (300 MHz, CDC13) δ 7.40 (IH, s), 7.27 - 7.32 (3H, m), 5.40 (IH, dd, J = 3.2, 9.5 Hz), 4.51 (IH, dd, J = 9.5, 12.1 Hz), 4.36 (IH, dd, /= 3.3, 12.1 Hz), 0.85 (9H, t, J = 7.5 Hz), 0.54 (6H, q, J = 7.5 Hz).
Figure imgf000043_0002
2-(3-Chloro-phenyl)-2-triethylsilanyloxy-ethylamine: Raney nickel (1 g) was washed with distilled water five times and methanol three times. [l-(3-Chloro-phenyl)-2- nitro-ethoxy]-triethyl-silane (10 g, 0.032 mol) and Raney nickel in methanol (100 mL) was hydrogenated (35 psi) at room temperature for 14 h. The reaction mixture was filtered through a pad of celite and rinsed with methanol. Concentration of the filtrate to dryness gave the title compound (5.6 g, 62%) as a colorless oil which was used for the next step without purification. 1H NMR (300 MHz, CDC13) δ 7.32 (IH, s), 7.18-7.26 (3H, m), 4.70 (IH, t, /= 5.8 Hz), 2.86 (2H, m), 0.89 (9H, t, /= 7.9 Hz), 0.56 (6H, q, /= 7.8 Hz). LRMS (M+H)+ m/z 286.
General Procedure for the Preparation of 2-Hydroxy-2-(substituted-phenyl)- ethylamines:
Figure imgf000044_0001
4-methoxy-3-bromophenyl chloroacetophenone: To a suspension of A1C13 (13.4g, 0.10 mol) in methylene chloride (40mL) was added a solution of 2-bromoanisole (12.5 mL, 0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution was warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washed with saturated sodium bicarbonate solution, brine, and dried over MgSO4. The solution was filtered, concentrated and crystalized from EtOH to give 15.37g of white solid. LRMS [M-H]- 260.8; IR (KBr) 1697, 1048, 1255 cm"1; 1H NMR (300MHz, CDC13) δ 8.18 (s, IH), 7.94 (dd, J= 8.67 Hz, IH), 6.96 (d,
J=8.67 Hz, IH), 4.62 (s, 2H), 3.98 (s, 3H); BC NMR (CDC13, 75.5 Hz) δ 188.8,
160.3, 134.1, 129.9, 128.2, 112.4, 111.3, 56.6, 45.3.
General Procedure for chiral reduction of chloroketones and ammonolysis :
OH
Figure imgf000044_0002
(S)-l-[4-methoxy-3-bromophenyl] -2-chloro ethanol : To a solution of (S)-Methyl- CBS-oxazaborolidine (1M in toluene, 0.745 mL, 0.745 mmol) and BH3-THF (8mL, 8 mmol) was added at the same time a solution of BH3-THF (19mL, 19 mmol) and a solution of the chloroketone (10.03g, 37.98 mmol) in 19 mL of THF. Both solutions were added dropwise over 30 minutes. The solution was stirred for 1 hour and quenched with the slow addition of methanol (50mL). The solution was concentrated and the residue chromatographed over a short silica gel column (1:1 hexane/ethyl acetate) to give a quantitative yield (lO.Og) of chlorohydrin as a clear oil. IR (KBr) 1053, 1258, 3406 cm"1; H NMR (300MHz, CDC13) δ 7.59 (s, IH), 7.30 (dd, J=2.16
Hz, IH), 6.90 (d, J=8.46 Hz, IH), 4.83 (dd, J= 3.57 Hz, IH), 3.90 (s, 3H), 3.64 (ddd,
J= 3.6, 11.1, 8.7, 2H), 2.04 (b s, IH). °C NMR (CDC13, 75.5 MHz) δ 155.9, 133.5,
131.1, 126.3, 111.9, 73.1, 60.4, 56.3, 50.7. OH
Figure imgf000045_0001
(S) 2-Amino-l-[3-chloro-4-methoxyphenyl]ethanol Hydrochloride : To a solution of the chlorohydrin (lO.Og, 37.9 mmol) in 120 mL of methanol at - 0°C was added 100 grams of ammonia. The solution was sealed in a pressure bottle and warmed to ambient temperature and stirred for 48 hours. The solution was cooled and opened. The ammonia was allowed to evaporate and solution concentrated. The residue was crystalized from ethanol/ethyl acetate to give 3.83g of white solid (35%). The material was reacted with B0C2O in ethyl acetate and saturated sodium bicarbonate and analyzed by chiral HPLC using a chiralcel OJ column using 95% hexane/ethanol as elutant and determined to by 98%ee. Additional crops were collected - 2.96g and 1.41g for a total of 75% yield. LRMS [M+H]+ 246; IR (cm"1, KBr) 1055, 1261,
3001, 2948, 3356; H NMR (500MHz, DMSO) δ 8.09 (b s, 2 H), 7.58 (s, IH), 7.36
(dd, J=2.05, 6.45 Hz, IH), 7.11 (d, J=8.5 Hz, IH) 6.10 (s, IH), 4.80 (m, IH), 3.84 (s, 3 3HH)),, 33..0000 ((dddddd,, JJ== 22..77,, 1122..66,, 99..55 HHzz,, 22HH));; 1133CC N NMMIR (DMSO, 75.5 MHz) δ 154.8, 135.4, 130.4, 126.6, 112.4, 110.4, 67.9, 56.2, 45.4.
OH
..NH2-HCI
(S) 2-Amino-l-[3-chlorophenyl]ethanol Hydrochloride : was prepared according to the general procedure outlined above. LRMS [M+H]+ 172; IR (KBr, cm-1) 3048,
3351, 2952; *H NMR (300MHz, MeOD) δ 7.48 (s, IH), 7.35 (m, 3H), 3.31 (ddd, J= 1.5, 3.12, 9.15 Hz 2H).
OH
Figure imgf000045_0002
(S)-2- Amino- 1- [3 -bromophenyl] ethanol Hydrochloride : was prepared according to the general procedure outlined above. LRMS [MH]+ 217.9; IR (KBr, cm-1) 3025, 3443, 2891; *H NMR (500MHz, DMSO) δ 7.93 (b s, 2H), 7.60 (s, IH), 7.52 (d, IH), 7.41 (s, IH), 7.35 (d, J=7.7 Hz, IH) 6.17 (s, IH), 4.82 (m, IH), 3.08 (ddd, J= 2.6, 12.7, 9.6 Hz, 2H); "c NMR (DMSO, 75.5 MHz) δ 144.4, 130.5, 128.7, 125.0, 121.6, 68.3, 45.1.
OH
,NH2-HCI
MeS ΛJ
(S)-2-Amino-l-[3-chloro-4-methylthiophenyl]ethanol Hydrochloride : was prepared according to the general procedure outlined above. LRMS [M+H]+ 217.9; IR (KBr, cm-1) 3007, 3358; *H NMR (500MHz, DMSO) δ 8.12 (b s, 2H), 7.46 (s, IH), 7.37 (s, IH), 7.35 (d, IH) 6.19 (d, IH), 4.83 (m, IH), 3.01 (ddd, J= 3.2, 12.8, 9.3 Hz, 2H); 13C NMR (DMSO, 75.5 MHz) δ 139.6, 136.5, 129.8, 126.6, 125.4, 68.0, 45.2, 14.2.
.OH Cl. ^. .A. .NH2-HCI
(S)-2-Amino-l-[3-chloro-4-fluoro-phenyl]ethanol Hydrochloride : was prepared according to the general procedure outlined above. LRMS [M+H]+ 189.9; IR (KBr, cm-1) 1509, 3008, 3359; H NMR (500MHz, DMSO) δ 8.21 (b s, 2H), 7.61 (d,
J=7.85 Hz, IH), 7.42 (m, 2H), 6.29 (s, IH), 4.88 (m, IH), 3.03 (ddd, J= 3.4, 12.8, 9.2 H Hzz,, 22HH));; 1 B3CC NNMMRR ((DDJMV SO, 75.5 MHz) δ 157.5, 155.5, 139.7, 128.1, 126.7, 119.3,
116.7, 109.0, 67.8, 45.2.
Figure imgf000046_0001
(S)-2- Amino- l-[3-chloro-4-methoxyphenyl]ethanol Hydrochloride : was prepared according to the general procedure outlined above. LRMS [M+H]+ 202; IR (KBr, cm-1) 3354, 3003, 2949, 1288, 1064; H NMR (500MHz, DMSO) δ 8.18 (brs, 3H), 7.43 (d, J=2.0Hz, IH), 7.31 (dd, J=8.5, 2.0Hz, IH), 7.14 (d, J=5.1Hz, IH), 6.11 (s, IH), 4.81 (m, IH), 3.84 (s, 3H), 2.99 (dd, J=13, 3.5Hz, IH), 2.83 (dd, J=12.5, 9Hz,
IH); BC NMR (DMSO, 125MHz) δ 153.9, 135.0, 127.3, 125.8, 120.8, 112.6, 68.0, 56.1, 45.5; Elemental Analysis Calcd for C9H12C1N02-HC1: C, 45.39; H, 5.50; N, 5.88. Found: C, 45.38; H, 5.43; N, 5.70.
OH
Figure imgf000047_0001
(S)-2-Amino-l-(7-bromo-2,3-dihydrobenzfuran-5-yl)-2-aminoethanol Hydrochloride : was prepared according to the general procedure outlined above. LRMS [M+H]+
258; IR (KBr, cm-1) 3349, 3006, 2928, 1485, 1045, 983; H NMR (500MHz, DMSO) δ 8.13 (brs, 3H), 7.29 (s, IH), 7.23 (s, IH), 6.08 (d, J=4 Hz, IH), 4.76 (m, IH), 4.61 (t, J=9Hz, 2H), 3.29 (t, J=9Hz, 2H), 2.96 (dd, J=13, 3.5Hz, IH), 2.82 (dd, J=13,9.5Hz, IH); 13C NMR (DMSO, 125MHz) δ 156.3, 135.9, 129.1, 128.1, 122.1, 100.9, 71.5, 68.2, 45.6, 29.9; Elemental Analysis Calcd for C10H12BrNO2-HCl: C, 40.77; H, 4.44; N, 4.75. Found: C, 40.77; H, 4.63; N, 4.63.
General Procedure for the Preparation of 2-Amino-3-(substituted-phenyl)-propanol:
Figure imgf000047_0002
(S)-[2-(3-Bromo-phenyl)-l-hydroxymethyl-ethyl]-carbamic acid tert-butyl ester: To a solution of (S)-3-(3-bromo-phenyl)-2-tert-butoxycarbonylamino-propinic acid (500 mg, 1.45 mmol) in THF (30 mL) was added borane-tetrahydrofuran complex (1.0 M solution) (4.35 mL, 4.35 mmol). The reaction mixture was stirred at room temperature for 14 h and quenched with acetic acid (1 mL). After removal of most solvent, the residue was extracted with EtOAc, washed with brine, dried over Na2SO4. After concentration, the crude product (400 mg, 83%) was used for the next step without purification. LCMS (M+H)+ m/z 330 (t = 1.61 min
Figure imgf000048_0001
(S)-2-Amino-3-(3-bromo-phenyl)-propan-l-ol: To a solution of (S)-[2-(3-bromo- phenyl)-l-hydroxymethyl-ethyl]-carbamic acid tert-butyl ester (400 mg, 1.21 mmol) in methanol (30 mL) was added 4 M HC1 in dioxane (2 mL, excess). The reaction mixture was stirred at room temperature for 14 h. After concentration in vacuo, the residue was used for the next step without purification. LCMS (M+H)+ m/z 230 (t = 0.78 min.)
Figure imgf000048_0002
5-Chloroacetyl-7-chlorooxindole: To a suspension of A1C13 (13.4g, 0.10 mol) in methylene chloride (40mL) is added a solution of 7-Chlorooxindole (0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution is warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washing with saturated bicarbonate solution, brine, and drying over MgSO4 would provide the desired chloroketone.
Figure imgf000048_0003
(S)-7-Chloro-5-(2-chloro-l-hydroxy-ethyl)-2-oxoindole : To a solution of (S)-Methyl- CBS-oxazaborolidine (1M in toluene, 0.745 mL, 0.745 mmol) and BH3-THF (8mL, 8 mmol) is added at the same time a solution of BH3-THF (19mL, 19 mmol) and a solution of the 5-Chloroacetyl-7-chlorooxindole (37.98 mmol) in 19 mL of THF. Both solutions are added dropwise over 30 minutes. The solution is stirred for 1 hour and quenched with the slow addition of methanol (50mL). The solution is concentrated and the residue chromatographed over a short silica gel column (1:1 hexane/ethyl acetate).
Figure imgf000049_0001
(S)-2-Amino-l-(7-chlorooxindole-5-yl)-ethanol Hydrochloride : To a solution of the chlorohydrin (37.9 mmol) in 120 mL of methanol at -40°C is added 100 grams of ammonia. The solution is sealed in a pressure bottle and warmed to ambient temperature and stirred for 48 hours. The solution is cooled and opened. The ammonia is allowed to evaporate and solution concentrated to provide the hydrochloride salt, which can be crystallized from ethanol ethyl acetate.
Figure imgf000049_0002
6-Chloroacetyl-4-chloro-2-benzooxazolinone: To a suspension of A1C13 (13.4g, 0.10 mol) in methylene chloride (40mL) is added a solution of 4-chloro-2- benzooxazolinone (0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution is warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washing with saturated bicarbonate solution, brine, and drying over MgSO4 would provide the desired chloroketone.
Figure imgf000049_0003
(S)-6-(2-Chloro-l-hydroxy-ethyl)-4-chloro-2-benzooxazolinone: To a solution of (S)- Methyl-CBS-oxazaborolidine (IM in toluene, 0.745 mL, 0.745 mmol) and BH3-THF (8mL, 8 mmol) is added at the same time a solution of BH3-THF (19mL, 19 mmol) and a solution of the 6-Chloroacetyl-4-chloro-2-benzooxazolinone (37.98 mmol) in 19 mL of THF. Both solutions are added dropwise over 30 minutes. The solution is stirred for 1 hour and quenched with the slow addition of methanol (50mL). The solution is concentrated and the residue chromatographed over a short silica gel column (1:1 hexane/ethyl acetate).
OH
Figure imgf000050_0001
(S)-2- Amino- l-(4-chloro-2-benzooxazolinone-6-yl)-ethanol Hydrochloride : To a solution of the chlorohydrin (37.9 mmol) in 120 mL of methanol at - 0°C is added 100 grams of ammonia. The solution is sealed in a pressure bottle and warmed to ambient temperature and stirred for 48 hours. The solution is cooled and opened. The ammonia is allowed to evaporate and solution concentrated to provide the hydrochloride salt, which can be crystallized from ethanol/ethyl acetate.
Figure imgf000050_0002
N-Methyl-7-chloroindoline: To a solution of the 7-Chloroindoline (0.10 mol) in 500 mL of acetone is added K2CO3 (0.15mol) and Mel (0.15mol) and refluxed until the starting material is consumed. The reaction is filtered and washed with water and saturated bicarbonate solution, drying over MgSO4 would provide the N-Me -7- chloroindoline
Figure imgf000050_0003
N-Methyl-5-chloroacetyl-7-chloro-indoline: To a suspension of A1C13 (13.4g, 0.10 mol) in methylene chloride (40mL) is added a solution of N-Me-7-Chloroindoline (0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution is warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washing with saturated bicarbonate solution, brine, and drying over MgSO4 provides the desired chloroketone.
OH
Figure imgf000051_0001
(S)-N-Methyl-5-(2-chloro-l-hydroxylethyl)-7-chloro-indoline: To a solution of (S)- Methyl-CBS-oxazaborolidine (IM in toluene, 0.745 mL, 0.745 mmol) and BH3-THF (8mL, 8 mmol) is added at the same time a solution of BH3-THF (19mL, 19 mmol) and a solution of N-Methyl-5-chloroacetyl-7-chloro-indoline (37.98 mmol) in 19 mL of THF. Both solutions are added dropwise over 30 minutes. The solution is stirred for 1 hour and quenched with the slow addition of methanol (50mL). The solution is concentrated and the residue chromatographed over a short silica gel column (1:1 hexane/ethyl acetate).
Figure imgf000051_0002
(S)-2-Amino-l-(7-chloro-N-methyl-indoline-5-yl)-ethanol Hydrochloride : To a solution of the chlorohydrin (37.9 mmol) in 120 mL of methanol at -40°C is added 100 grams of ammonia. The solution is sealed in a pressure bottle and warmed to ambient temperature and stirred for 48 hours. The solution is cooled and opened. The ammonia is allowed to evaporate and solution concentrated to provide the hydrochloride salt, which can be crystallized from ethanol/ethyl acetate.
Figure imgf000051_0003
(S)-2-Chloro-l-(7-chloro-N-methyl-indol-5-yl)-ethanol: A solution (S)-N-Methyl-5- (2-chloro-l-hydroxylethyl)-7-chloro-indoline (0.10 mmol) in lOOmL of t-butyl methyl ether is treated with o-chloroanil (O.lOmmol) at ambient temperature. The solution is concentrated and the residue chromatographed over silica gel (1:1 hexane/ethyl acetate) to provide the corresponding indole.
Figure imgf000052_0001
(S)-2-Amino-l-(7-chloro-N-methyl-indol-5-yl)-ethanol Hydrochloride : To a solution of the chlorohydrin (37.9 mmol) in 120 mL of methanol at -40°C is added 100 grams of ammonia. The solution is sealed in a pressure bottle and warmed to ambient temperature and stirred for 48 hours. The solution is cooled and opened. The ammonia is allowed to evaporate and solution concentrated to provide the hydrochloride salt, which can be crystalized from ethanol/ethyl acetate.
Figure imgf000052_0002
4-Chloro-2-methyl-benzooxazole : To a solution of the 4-chloro-2-benzooxazolinone (0.10 mol) in 200 mL ethanol is added LiOH (0.20 mol) in 100 mL of water. The solution is refluxed for 8 hr and cooled. The solution is neutralized with IN HC1 and extracted with ethyl acetate followed by drying over MgSO4. The solution is concentrated and taken up in 200 mL of toluene and 0.10 mol of acetic acid. The solution is refluxed in a Dean Stark trap for 12 hours, concentrated and purified by flash chromatography.
Figure imgf000052_0003
6-Chloroacetyl-4-chloro-2-methyl-benzooxazole: To a suspension of A1C13 (13.4g, 0.10 mol) in methylene chloride (40mL) is added a solution of 2-Methyl-4-chloro- benzooxazole (0.10 mol) and chloroacetyl chloride (8 mL, 0.10 mol) at 0°C. The solution is warmed to ambient temperature for two hours and poured onto ice and extracted with methylene chloride, washing with saturated bicarbonate solution, brine, and drying over MgSO4 would provide the desired chloroketone.
Figure imgf000053_0001
(S )-6-(2-chloro- 1 -hydroxy-ethyl)-4-chloro-2-methyl-benzooxazole : To a solution of (S)-Methyl-CBS-oxazaborolidine (IM in toluene, 0.745 mL, 0.745 mmol) and BH3- THF (8mL, 8 mmol) is added at the same time a solution of BH3-THF (19mL, 19 mmol) and a solution of 6-Chloroacetyl-4-chloro-2-methyl-benzooxazole (37.98 mmol) in 19 mL of THF. Both solutions are added dropwise over 30 minutes. The solution is stirred for 1 hour and quenched with the slow addition of methanol
(50mL). The solution is concentrated and the residue chromatographed over a short silica gel column (1:1 hexane/ethyl acetate).
Figure imgf000053_0002
(S)-2-Amino-l-(4-chloro-2-methyl-benzooxazol-6-yl)-ethanol Hydrochloride : To a solution of the chlorohydrin (37.9 mmol) in 120 mL of methanol at -40°C is added 100 grams of ammonia. The solution is sealed in a pressure bottle and warmed to ambient temperature and stirred for 48 hours. The solution is cooled and opened. The ammonia is allowed to evaporate and solution concentrated to provide the hydrochloride salt, which can be crystallized from ethanol/ethyl acetate. Preparation of 3-(6-Imidazol- 1 -yl-4-methyl- 1 H-benzimidazol-2-yl)-4-iodo- 1 H- pyridin-2-one:
Figure imgf000054_0001
l-(3-Methyl-4-nitro-phenyl)-lH-imidazole: To a solution of 4-fluoro-2-methyl-l- nitro-benzene (300 mg, 1.84 mmol) in DMSO (2 mL) were added KOH (20 mg, 3.87 mmol) and imidazole (263 mg, 3.88 mmol). The reaction mixture was heated to 100°C for 3.5 h, cooled to room temperature, and diluted with ice-cold water. The resulting precipitate was filtered, washed with ice-cold water, and dried under vacuum to give the title compound (310 mg, 80%) as a yellow powder. 1H NMR (300 MHz, DMSO-d6) δ 8.46 (IH, s), 8.16 (IH, d, J = 8.9 Hz), 7.90 - 7.92 (2H, m), 7.78 (IH, dd, 7= 2.5, 8.9 Hz), 7.17 (IH, s), 2.61 (3H, s). LRMS (M+H)+ m/z 204.
Figure imgf000054_0002
4-Imidazol-l-yl-2-methyl-phenylamine: To l-(3-methyl-4-nitro-phenyl)-lH- imidazole (200 mg, 0.98 mmol) and 10% Palladium on carbon (35 mg) was added degassed methanol (3 mL). The suspension was flushed and evacuated with hydrogen / vacuum line. The suspension was allowed to stir at room temperature for 14 h under hydrogen atmosphere (hydrogen balloon). The dark reaction mixture was filtered through a pad of celite and rinsed with methanol. Concentration of the filtrate gave the title compound (166 mg, 98%) which was used for the next step without purification. 1H NMR (300 MHz, DMSO-de) δ 7.95 (IH, s), 7.48 (IH, s), 7.16 (IH, narrow d, / = 2.5 Hz), 7.09 (IH, dd, /= 2.5, 8.4 Hz), 7.01 (IH, s), 6.67 (IH, d, /= 8.4 Hz), 5.03 (2H, broad s), 2,10 (3H, s).
Figure imgf000055_0001
N-(4-Imidazol-l-yl-2-methyl-6-nitro-phenyl)-acetamide: To a solution of 4-imidazol- l-yl-2-methyl-phenylamine (1 g, 5.78 mmol) in CH2C12 (20 mL) was added Ac2O (0.7 mL, 7.28 mmol) at 0°C. The reaction mixture was stir at room temperature for 14 h and diluted with water. The aqueous layer was extracted with CH2CI2 and the combined organic layers were washed with saturated ΝaHCO3 and brine, dried over Na2SO4, and concentrated in vacuo to give a white solid. The white solid was suspended in H2SO4 (cone.) (15 mL). Then HNO3 (cone.) (0.312 mL) was added to the suspension at 0°C. The reaction mixture was slowly warmed to room temperature and stirred at room temperature for 4 h. After cooling to -10°C, the reaction mixture was neutralized with ammonium hydroxide and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated. The residue was purified by flash chromatography (1:9:5 MeOH / THF / hexane) to yield the title compound (0.61 g, 41%). 'H NMR (300 MHz, CD3OD) δ 8.11 (IH, s), 7.45 - 7.56 (2H, m), 7.38 (IH, dd, J = 2.4, 8.4 Hz), 7.14 (IH, s), 2.33 (3H, s), 2.18 (3H, s).
Figure imgf000055_0002
4-Imidazol-l-yl-2-methyl-6-nitro-phenylamine: To a suspension of N-(4-imidazol-l- yl-2-methyl-6-nitro-phenyl)-acetamide (279 mg, 1.07 mmol) in ethanol (3 mL) was added 2 Ν HC1 (2 mL). The reaction mixture was heated to reflux for 14 h, cooled to room temperature, and neutralized with saturated ΝaHCO3. The resulting bright orange solid was filtered and dried under vacuum. The title compound (179 mg, 76%) was obtained as an orange solid. 1H NMR (300 MHz, CD3OD) δ 8.78 (IH, s), 8.24 (IH, s), 7.78 (IH, s), 7.64 (IH, s), 7.46 (IH, s), 2.36 (3H, s).
Figure imgf000056_0001
5-Imidazol-l-yl-3-methyl-benzene-l,2-diamine: To 4-imidazol-l-yl-2-methyl-6-nitro- phenylamine (350 mg, 1.61 mmol) and 10% Palladium on carbon (40 mg) were added degassed methanol (5 mL) and TFA (5 drops). The reaction mixture was flushed and evacuated with hydrogen / vacuum line, stirred at room temperature for 14 h under hydrogen atmosphere (hydrogen balloon). The dark reaction mixture was filtered through a pad of celite and rinsed with methanol. Concentration of the filtrate gave the residue, which was diluted with water, extracted with ethyl acetate. The combined organic layers were washed with saturated NaHCO3, brine, dried over Na2SO4.
Concentration to dryness gave the title compound (275 mg, 91%) as a solid. 1H NMR (300 MHz, CD3OD) δ 7.87 (IH, s), 7.34 (IH, s), 7.05 (IH, s), 6.72 (IH, d, J = 2.4 Hz), 6.65 (IH, d, J= 2.4 Hz) 2.21 (3H, s). LCMS (M+H)+ m/z 189 (t = 0.23 min.).
Figure imgf000056_0002
4-Iodo-2-methoxy-pyridine-3-carbaldehyde (WO 95/29917): A 5-liter three-necked round bottom flask was equipped with an overhead mechanical stirrer under nitrogen, the flask was charged with THF (1 L) and cooled to -78°C. To this stirred solution was added tert-butyllithium (1.7 M solution in pentane) (800 mL, 1.36 mol) via canula followed by 2-methoxypyridine (132.2 g, 1.21 mol) at -78°C. The mixture was stirred for 1 h at -78°C. To the mixture was added N-formyl-N, N', N'- trimethylethylenediamine (176 mL, 1.37 mol) dropwise at -78°C. The reaction mixture was stirred for ca. 30 min at -78°C before warming to -23°C over ca. 30 min. To the mixture at -23°C was added ethylene glycol dimethyl ether (1 L) followed by n-butyllifhium (2.5 M solution in hexane) (800 mL, 2.0 mol). The resulting mixture was stirred for ca. 2 h during which time the reaction mixture turned deep green. A 12-L 4-necked round flask was charged with iodine (571 g, 2.25 mol) and ethylene glycol dimethyl ether (2 L) and the resultant solution was cooled to -78°C. The contents of the 5-L flask were transferred via canula to the mixture of iodine and ethylene glycol dimethyl ether in the 12-L flask at -78°C. After the addition was complete, the reaction mixture was stirred for an additional 1 h at -78°C. The cooling bath was removed and the mixture was allowed to warm to about 0°C and treated with 2 L of water and 2 L of 1 N hydrochloric acid. Methyl t-butyl ether (2L) was added and the layers were separated. The aqueous layer was extracted with 2 x 1 L of methyl t-butyl ether. The combined organic layers were washed with saturated Na2S2θ3 (1.2 L), brine (1.2 L), dried over Na2SO4. After concentration in vacuo, the thick slurry was diluted with hexane (1 L). The mixture was cooled with an ice / water bath for ca. 30 min. The precipitate was filtered and dried in vacuum to yield the title compound as a light yellow solid. 1H NMR (300 MHz, CDC13) δ 10.22 (s, IH), 7.86 (IH, d, J = 5.3 Hz), 7.54 (IH, d, J = 5.3 Hz), 4.06 (3H, s). LCMS (M+H)+ m/z 364 (t = 2.26 min.).
Figure imgf000057_0001
6-Imidazol-l-yl-2-(4-iodo-2-methoxy-pyridin-3-yl)-4-methyl-lH-benzimidazole: To a solution of 5-imidazol-l-yl-3-methyl-benzene-l,2-diamine (175 mg, 0.93 mmol) in methanol (8 mL) was added a solution of 4-iodo-2-methoxy-pyridine-3-carbaldehyde (245 mg, 0.93 mmol) in methanol (5 mL) at 0°C. The reaction mixture was stirred at 0°C for 1.5 h and then at room temperature for 2 h. After concentration, the residue was purified by flash column chromatography (10% MeOH / CH2CI2) to give the title compound (291 mg, 60%). 1H NMR (300 MHz, CD3OD) δ 8.13 (IH, s), 7.98 (IH, d, 7= 5.4 Hz), 7.62 (IH, d, / = 5.4 Hz), 7.59 (2H, s), 7.33 (IH, s), 7.16 (IH, s), 3.90 (3H, s), 2.67 (3H, s). LCMS (M+H)+ m/z 432 (t = 0.99 min.).
Figure imgf000057_0002
3-(6-Imidazol- 1 -yl-4-methyl- 1 H-benzimidazol-2-yl)-4-iodo- 1 H-pyridin-2-one: The suspension of 6-imidazol-l -yl-2-(4-iodo-2-methoxy-pyridin-3-yl)-4-methyl-lH- benzimidazole in 1 N HC1 (6 mL) was heated to 70°C for 3 days, cooled to room temperature, and diluted with ethyl acetate. After extraction, the combined organic layers were washed with brine, dried over Na2SO4 and concentrated. The residue was purified by flash chromatography (1% NHUOH / 10% MeOH / CH2C12) to yield the title compound (78 mg, 81%) as a solid. 1H NMR (300 MHz, CD3OD) δ 8.12 (IH, s), 7.58 (2H, s), 7.29 - 7.31 (2H, m), 7.16 (IH, s), 7.01 (IH, J = 6.8 Hz), 2.66 (3H, s). LCMS (M+H)+ m/z 418 (t = 0.75 min.)
Preparation of 2-(4-Chloro-2-oxo- 1 ,2-dihydro-pyridin-3-yl)-7-methyl-3H- benzimidazole-5-carbonitrile:
Figure imgf000058_0001
4-Amino-3-methyl-benzonitrile: To a solution of 3-methyl-4-nitro-benzonitrile (20 g, 0.123 mol) in HOAc (200 mL) was added iron powder (17.55 g, 0.309 mol). After 10 min, the reaction was exothermic and turned to dark color. The reaction mixture was allowed to stir at room temperature for 14 h and then diluted with EtOAc (200 mL). The brown precipitate was filtered through a pad of celite and the filtercake was rinsed with EtOAc. The filtrate was concentrated in vacuo and the residue was purified by flash chromatography (40% EtOAc / hexane) to yield the title compound (15.3 g, 92%). 1H NMR (300 MHz, CDC13) δ 7.30 - 7.34 (2H, m), 6.64 (IH, d, / = 8.7 Hz), 2.16 (3H, s). LCMS (M+H)+ m/z 133 (t = 0.93 min).
Figure imgf000058_0002
N-(4-Cyano-2-methyl-6-nitro-phenyl)-2,2,2-trifluoro-acetamide: To the ice-cold trifluoroacetic anhydride (60 mL) was added 4-amino-3-methyl-benzonitrile (14.33 g, 0.108 mol) in portion. The resulting white slurry was stirred at 0°C for 30 min. Then ammonium nitrate (17.28 g, 0.216 mol) was added. The reaction mixture was allowed to stir at 0°C for 1 h and at room temperature for 14 h. After removal of most solvent, the reaction mixture was cooled with ice and quenched with ice. The yellow precipitate was filtered, washed with cold water, and dried under vacuum. The crude product (15.5 g, 52% yield, and ca. 80% pure) was used for the next step without purification. *H NMR (300 MHz, CD3OD) δ 8.05 (IH, s), 7.74 (IH, s), 2.30 (3H, s). LRMS (neg. ESI, (M-H)") m z 272.
Figure imgf000059_0001
4-Amino-3-methyl-5-nitro-benzonitrile: A mixture of N-(4-cyano-2-methyl-6-nitro- phenyl)-2,2,2-trifluoro-acetamide (5 g, 18.3 mmol) and 2 M ammonia in methanol (80 mL) was heated to reflux for 14 h and then cooled to room temperature. After concentration in vacuo, the residue was purified by flash chromatography (20%
EtOAc / hexane) to yield the title compound (3.24 g, 100%, ca 80% pure). 1H ΝMR
(300 MHz, CDC13) δ 8.40 (IH, s), 7.47 (IH, s), 6.6 - 6.8 (2H, broad s), 2.89 (3H, s).
Figure imgf000059_0002
3,4-Diamino-5-methyl-benzonitrile: To a solution of 4-amino-3-methyl-5-nitro- benzonitrile (3.24 g, 18.3 mmol) in ethanol (80 mL) was added tin chloride dihydrate (8.67 g, 49.75 mmol). The reaction mixture was heated to reflux for 14 h, then cooled to room temperature, and concentrated in vacuum. The residue was diluted with ethyl acetate (100 mL) and treated with triethylamine (20 mL). The resulting slurry was filtered through a pad of celite and the filtercake was rinsed with three-portion ethyl acetate (50 mL). The filtrate was washed with saturated NaHCO3, water, and brine, then dried over Na2SO4 and filtered. After removal of solvent, the residue was purified by flash chromatography on silica gel (30% - 50% EtOAc / hexane) to yield the title compound (2.17 g, 81%) as a light yellow solid. 1H NMR (300 MHz, CDC13) δ 6.94 (IH, s), 6.85 (IH, s), 2.16 (3H, s). LCMS (M+H)+ m/z 148 (t = 0.67 min.).
Figure imgf000060_0001
2-(4-Iodo-2-methoxy-pyridin-3-yl)-7-methyl-3H-benzimidazole-5-carbonitrile: To a solution of 3,4-diamino-5-methyl-benzonitrile (2.00 g, 13.6 mmol) in MeOH (40 L) was added 4-iodo-2-methoxy-pyridine-3-carbaldehyde (3.6 g, 13.6 mmol) in MeOH (20 mL) at 0°C. The resulting slurry was stirred at 0°C for 1 h. Iodine (1.73 g, 8.8 mmol) in MeOH (10 mL) was added dropwise via a dropping funnel to the reaction mixture at 0°C. The reaction mixture was stirred at room temperature for 14 h. After removal of MeOH, the residue was diluted with saturated Na2S2θ3 and extracted with EtOAc. The combined organic layers were washed with water and brine, dried over Na2SO4. The crude product was purified by flash column chromatography (3% MeOH / CH2C12) to yield the title compound ( 1.81 g, 46%). 1H NMR (300 MHz, CDC13) δ 7.90 (IH, s), 7.49 (IH, d, J = 5.4 Hz), 7.46 (IH, s), 7.41 (IH, d, J = 5.3 Hz), 3.78 (3H, s), 2.68 (3H, s). LCMS (M+H)+ m z 391 (t = 1.27 min.).
Figure imgf000060_0002
2-(4-Chloro-2-oxo-l,2-dihydro-pyridin-3-yl)-7-methyl-3H-benzimidazole-5- carbonitrile: A suspension of 2-(4-iodo-2-methoxy-pyridin-3-yl)-7-methyl-3H- benzimidazole-5-carbonitrile (1.8 g, 4.63 mmol) in 4 M HC1 dioxane (40 mL) was heated to 80°C for 6 h and cooled to room temperature. The precipitate was filtered and dried. The crude product (1.08 g, 82%) was used for the next step without purification. LRMS (neg. ESI, (M-H)") m/z 283. Preparation of (S)-4-(l-Benzyl-2-trityloxy-ethylamino)-3-(6-bromo-4-methyl-lH- benzimidazol-2-yl)-lH-pyridin-2-one:
Figure imgf000061_0001
5-Bromo-3-methyl-benzene-l,2-diamine: To a suspension of 4-bromo-2-methyl-6- nitro-phenylamine (20 g, 0.086 mol) in ethanol (200 mL) was added tin chloride dihydrate (49.2 g, 0.258 mol). The reaction mixture was heated to reflux for 14 h, cooled to room temperature, and concentrated in vacuo. The residue was diluted with ethyl acetate (150 mL) and treated with triethylamine (40 mL). The resulting slurry was filtered through a pad of celite, and the filtercake was rinsed with three portions ethyl acetate (50 mL). The filtrate was washed with saturated NaHCO3, water, and brine, then dried over Na2SO and filtered. After removal of the solvent, the residue was purified by flash chromatography on silica gel (30% EtOAc / hexane, then 5% MeOH / CH2C12) to yield the title compound (10.26 g, 59%) as a yellow oil. 1H NMR (300 MHz, CDC13) δ 6.77 (IH, d, J = 2.0 Hz), 6.74 (IH, d, /= 2.0 Hz), 2.16 (3H, s). LCMS (M+H)+ m/z 201. (t = 0.83 min.).
Figure imgf000061_0002
6-Bromo-2-(4-iodo-2-methoxy-pyridin-3-yl)-4-methyl- lH-benzimidazole: To a solution of 5-bromo-3-methyl-l,2-phenylenediamine (4 g, 19.9 mmol) in methanol (80 mL) was added 4-iodo-2-methoxy-pyridine-3-carbaldehyde (5.23 g, 19.9 mmol) in methanol (20 mL) dropwise at 0°C. The resulting slurry was stirred for 30 min at room temperature. Then iodine (2.53 g, 9.95 mmol) in methanol (20 mL) was added via a dropping funnel. After 14 h, the reaction mixture was concentrated in vacuo, diluted with 5% Na2S2θ3, and extracted with ethyl acetate. The combined organic layers were washed with water, brine, and dried over Na2SO4. After removal of solvent, the residue was purified by careful flash chromatography (20% EtOAc / hexane) to yield the title compound (4.05 g, 46%) as a yellow foam. !H NMR (300 MHz, CDC13) δ 7.86 (IH, d, J= 5.31 Hz), 7.53 (IH, d, J= 5.3 Hz), 7.26 (2H, broad s), 3.91 (3H, s), 2.63 (3H, s). LCMS (M+H)+ m z 444 (t = 1.39 min.).
Figure imgf000062_0001
3-(6-Bromo-4-methyl-lH-benzimidazol-2-yl)-4-chloro-lH-pyridin-2-one: The suspension of 6-bromo-2-(4-iodo-2-methoxy-pyridin-3-yl)-4-methyl-lH- benzimidazole (4 g, 9.03 mmol) and 60 mL of 4 M HC1 in dioxane was heated to 80°C for 6 h and cooled to room temperature. The precipitate was filtered and dried to yield the title compound (3.0 g, 100%) as a brown powder. The crude product was used for the next step without purification. Η NMR (300 MHz, CD3OD) δ 7.55 (IH, s), 7.42 (IH, d, J = 6.0 Hz), 7.17 (IH, s), 6.91 (IH, d, J= 6.0 Hz), 2.55 (3H, s). LC MS (M+H)+ m/z 338 (t = 1.33 min.).
Figure imgf000062_0002
(S)-4-(l-Benzyl-2-hydroxy-ethylamino)-3-(6-bromo-4-methyl-lH-benzimidazol-2- yl)-lH-pyridin-2-one: To a solution of 3-(6-bromo-4-methyl-lH-benzimidazol-2-yl)- 4-chloro-lH-pyridin-2-one (1.42 g, 3.78 mmol) in DMF (15 mL) were added (S)-(-)- 2-amino-3-phenyl-l-propanol (1.43 g, 9.45 mmol) and N-methyl moφholine (1.5 mL). The reaction mixture was heated to 80°C for 6 h and cooled to room temperature. The solvent was removed with high vacuum and the residue was purified by flash chromatography (5% MeOH / CH2CI2) to yield the title compound (1.26 g, 74%) as a yellow foam. 1H ΝMR (300 MHz, CDC13) δ 6.9 - 7.2 (8H, m), 5.86 (IH, d, J= 7.1 Hz), 3.7-3.9 (3H, m), 2.9-3.1(2H, m), 2.57 (3H, s). LCMS (M+H)+ m/z 453 (t = 2.03 min.).
Figure imgf000063_0001
(S)-4-(l-Benzyl-2-trityloxy-ethylamino)-3-(6-bromo-4-methyl-lH-benzimidazol-2- yl)-lH-pyridin-2-one: To a solution of (S)-4-(l-benzyl-2-hydroxy-ethylamino)-3-(6- bromo-4-methyl-lH-benzimidazol-2-yl)-lH-pyridin-2-one (0.9 g, 1.98 mmol) in THF (30 mL), was added Cs2CO3 (1.29 g, 3.96 mmol) followed by triphenylmethyl chloride (1.10 g, 3.96 mmol). The reaction mixture was heated to reflux for 14 h under nitrogen and then cooled to room temperature. After removal of the solvent, the residue was diluted with ethyl acetate and washed with water. The aqueous fraction was extracted with ethyl acetate and the combined organic layers were washed with water and brine, dried over Na2SO4. After concentration in vacuo, the residue was purified by flash column chromatography (30% EtOAc / hexane) to yield the title compound (1 g, 73%) as a white solid. 1H NMR (300 MHz, DMSO-d6) δ 11.77 (IH, broad s), 11.73 (IH, d, J = 5.2 Hz), 11.46 (IH, broad s), 7.13 - 7.54 (23H, m), 5.87 (IH, d, J = 4.5 Hz), 4.09 - 4.14 (IH, m), 3.07 - 3.42 (4H, m), 2.54 (3H, s). LCMS (M+H)+ m/z 695 (t = 2.79 min.).
Preparation of 3 - [6-(4- Acetyl-piperazin- 1 -yl)-4-methyl- 1 H-benzimidazol-2-yl] -4- chloro- 1 H-pyridin-2-one:
Figure imgf000063_0002
l-[4-(4-Amino-3-methyl-5-nitro-phenyl)-piperazin-l-yl]-ethanone: A mixture of 4-bromo- 2-methyl-6-nitro-phenylamine (5g, 21.64 mmol), 1-acetylpiperazine (4.2g, 32.46 mmol), palladium acetate (244 mg, 1.08 mmol), tri-tert-butylphosphine (440 mg, 2.16 mmol) and sodium tert-butoxide (4.2 g, 43.29 mmol) in toluene (70 mL) was heated to 100°C for 14 h under nitrogen. The reaction mixture was cooled to room temperature and diluted with EtOAc. After extraction, the combined organic layers were washed with water, brine, dried over Na2SO4. Concentration gave a brownish residue which was purified by flash column chromatography (10% MeOH / CH2C12) to yield the title compound (4.21 g, 70%). 1H NMR (400 MHz, CD3OD) δ 7.42 (IH, d, 7 = 2.8 Hz), 7.23 (IH, d, 7 = 2.8 Hz), 3.71 (2H, t, 7= 5.1 Hz), 3.67 (2H, t, 7 = 5.1 Hz), 3.04 (2H, t, 7 = 5.2 Hz), 2.98 (2H, t, 7= 5.2 Hz), 2.24 (3H, s), 2.13 (3H, s). LCMS (M+H)+ m/z 279 (t = 1.46 min.).
Figure imgf000064_0001
1 -[4-(3,4-Diamino-5-methyl-phenyl)-piperazin- 1 -yl]-ethanone: To 1 -[4-(4-amino-3- methyl-5-nitro-phenyl)-piperazin-l-yl]-ethanone (4.5 g, 16.2 mmol) and 10% palladium on carbon (400 mg) were added methanol (50 mL) and acetic acid (5 mL) under nitrogen. The reaction mixture was stirred under hydrogen atmosphere (hydrogen balloon) for 14 h. The dark solution was filtered through a pad of celite and the fϊltercake was washed with methanol. Concentration of the filtrate gave the title compound (4.00 g, 100%) which was used for the next step without purification. LCMS (M+H)+ m/z 207 (t = 0.41 min.).
Figure imgf000064_0002
1 - { 4- [2-(4-Iodo-2-methoxy-pyridin-3 -yl)-7-methyl-3H-benzimidazol-5-yl] -piperazin- 1 - yl}-ethanone: To a solution of l-[4-(3,4-diamino-5-methyl-phenyl)-piperazin-l-yl]- ethanone (4.00 g, 16.18 mmol) in methanol (100 mL) was added 4-iodo-2-methoxy- pyridine-3-carbaldehyde (4.25 g, 16.18 mmol). The reaction mixture was stirred at room temperature for 14 h. After concentration, the residue was purified by flash column chromatography (10% MeOH / CH2C12) to yield the title compound (5.25 g, 66%). 1H NMR (400 MHz, CD3OD) δ 7.81 (IH, d, 7 = 5.4 Hz), 7.48 (IH, d, 7 = 5.4 Hz), 7.26 (IH, s), 6.85 (IH, s), 3.85 (3H, s), 3.78 (2H, t, 7 = 5.0 Hz), 3.64 (2H, t, 7 = 5.0 Hz), 3.16 (2H, t, 7= 5.2 Hz), 3.11 (2H, t, 7 = 5.2 Hz), 2.62 (3H, s), 2.13 (3H, s). LCMS (M+H)+ m z 492 (t = 1.71 min.).
Figure imgf000065_0001
3 - [6-(4- Acetyl-piperazin- 1 -yl)-4-methyl- 1 H-benzimidazol-2-yl] -4-chloro- 1 H-pyridin- 2-one: To a solution of l-{4-[2-(4-Iodo-2-methoxy-pyridin-3-yl)-7-methyl-3H- benzimidazol-5-yl]-piperazin-l-yl}-ethanone (5.2 g, 10.6 mmol) in 4 M HC1 in dioxane (60 mL) was added water (5 mL). The reaction mixture was stirred at room temperature for 14 h. Concentration of the reaction mixture gave the title compound (4.02 g, 100%) which was used for the next step without purification. LCMS (M+H)+ m/z 486 (t = 1.55 min.).
Preparation of (S)-4-(l-Hydroxymethyl-2-phenyl-ethylamino)-3-(4-methyl-6-piperazin-l- yl- 1 H-benzimidazol-2-yl)- 1 H-pyridin-2-one :
Figure imgf000065_0002
(S)-3-[6-(4-Acetyl-piperazin-l-yl)-4-methyl-lH-benzimidazol-2-yl]-4-(l-hydroxymethyl- 2-phenyl-ethylamino)-lH-pyridin-2-one: To a solution of 3-[6-(4-acetyl-piperazin-l-yl)-4- methyl-lH-benzimidazol-2-yl]-4-chloro-lH-pyridin-2-one (1 g, 2.6 mmol) in DMF (10 mL) was added (S)-(-)-2-amino-3-phenyl-propanol (0.78 mg, 5.2 mmol) and N-methyl moφholine (2 mL). The reaction mixture was heated to 80°C for 12 h, cooled to room temperature and concentrated with high vacuum. The residue was purified by flash column chromatography (10% MeOH / CH2C12) to yield the title compound (0.90 g, 69%). 1H NMR (400 MHz, CD3OD) δ 7.36 (IH, s), 7.02 - 7.23 (6H, m), 6.80 (IH, s), 5.98 (IH, d, 7 = 7.5 Hz), 4.10 - 4.12 (3H, m), 3.67 - 3.78 (6H, m), 3.06 - 3.11 (3H, m), 2.90 (IH, dd, 7 = 7.8, 13.6 Hz), 2.54 (3H, s), 2.12 (3H, s). LCMS (M+H)+ m/z 501 (t = 1.30 min.).
Figure imgf000066_0001
(S)-4-( 1 -Hydroxymethyl-2-phenyl-ethylamino)-3-(4-methyl-6-piperazin- 1 -yl- 1 H- benzimidazol-2-yl)-lH-pyridin-2-one: To a solution of (S)-3-[6-(4-acetyl-piperazin-l- yl)-4-methyl- 1 H-benzimidazol-2-yl]-4-( 1 -hydroxymethyl-2-phenyI-ethylamino)- 1 H- pyridin-2-one (900 mg, 18 mmol) in 4 M HC1 in dioxane (10 mL) was added water (1 mL). The reaction mixture was heated to 80°C for 14 h and cooled to room temperature. Concentration with high vacuum gave the title compound (0.83 g, 100%) which was used for the next step without purification. LCMS (M+H)+ m z 459 (t = 1.13 min.).
Preparation of 2-(4,6-Dichloro-pyrimidin-5-yl)-6-imidazol- 1 -yl-4-methyl- 1H- benzimidazole:
Figure imgf000066_0002
4,6-Dichloro-pyrimidine-5-carbaldehyde: DMF (7 mL, 0.09 mol) was added to POCl3 (21 mL, 0.23 mol) at 0°C. The reaction mixture was stirred at room temperature for 0.5 h. 4,6-Dihydroxy-pyrimidine-5-carbaldehyde (5 g, 0.045 mol) was added in small portions. The reaction mixture was heated to 90°C for 6 h and cooled to room temperature. A large excess of crushed ice was added to the reaction mixture very slowly under ice-bath. The mixture was extracted with CH2CI2. The combined organic layers were washed with water, brine, and dried over Na2SO4. After concentration, the residue was purified by column chromatography (20% EtOAc / hexane) to yield the title compound (4 g, 50%). 1H NMR (400 MHz, CDC13) δ 8.91 (IH, s), 7.87 (IH, s). LRMS (M+H)+ m/z 177.
Figure imgf000067_0001
2-(4,6-Dichloro-pyrimidin-5-yl)-6-imidazol-l-yl-4-methyl-lH-benzimidazole: To a solution of 5-imidazol-l-yl-3-methyl-benzene-l,2-diamine (180 mg, 0.96 mmol) in methanol (4 mL) was added a solution of 4,6-dichloro-pyrimidine-5-carbaldehyde ( 183 mg, 0.96 mmol) in methanol (1 mL). The reaction mixture was stirred at room temperature for 14 h. After concentration, the residue was purified by flash column chromatography (5% methanol / CH2CI2) to yield the title compound (180 mg, 55%). LCMS (M+H)+ m z 344 (t = 1.31 min.).
Procedure for the Preparation of 2-Amino-4-Fluoro-6-methyl nitrobenzene:
Figure imgf000067_0002
2-(3,5-Difluoro-2-nitro-phenyl)-malonic acid di-tert-butyl ester: To a suspension of NaH (54.6g, 60%, 1.365 mol) in 600mL of DMF was added di-t-Butyl malonate (118g, 0.546 mol) at 0°C and stirred for 30 min. 2,4,6 trifluoronitrobenzene was added as a solution in 400 mL of DMF (75 g, 0.42mol) over 3 hours a d the solution stirred at ambient temperature for 12 hours. The reaction mixture was extracted with ethyl acetate (3X's). The ethyl acetate was washed with water (3X's) and with brine and dried over MgSO and concentrated to give 62 g of crude product. LCMS [M+Na] - 396; 1H NMR (500MHz, DMSO) δ 7.81 (m, IH), 7.27 (m, IH), 5.00 (s, IH), 1.41 (m, 18H).
Figure imgf000068_0001
2-(3-Amino-5-fluoro-2-nitro-phenyl)-malonic acid di-tert-butyl ester: To the crude 2- (3,5-Difluoro-2-nitro-phenyl)-malonic acid di-tert-butyl ester (62g, 0.42 mol) was added 700 mL of 2M ammonia in methanol in a pressure bottle. The vessel was sealed and heated to 85°C for 18 hours. The reaction mixture was cooled and the vessel opened carefully and the methanol solution concentrated to provide 140 g of crude material. LCMS [M+Na] - 393; 1H NMR (500MHz, DMSO) δ 6.76 (dd, J=10.8 2.8Hz, IH), 6.29 (dd, J=10.8, 2.8Hz, IH), 4.99 (brs, 2H), 4.80 (s, IH), 1.40 (m, 18H).
Figure imgf000068_0002
3-Amino-5-fluoro-2-nitro phenyl acetic acid: To the 2-(3-Amino-5-fluoro-2-nitro- phenyl)-malonic acid di-tert-butyl ester (140g) in 500 mL of 4N HCl in dioxane was added 50 mL of water and heated to 40°C for 2 days. The solution was extracted with ethyl acetate (3X's) and the ethyl acetate washed with water (3X's) and brine. The organic fraction was dried over MgSO4 and was concentrated to give 78g of crude (66% pure by LC/MS); *H NMR (500MHz, DMSO) δ 12.40 (brs, IH), 7.04 (s, 2H), 6.68 (dd, J=10.9 2.8Hz, IH), 6.47 (dd, J=10.9, 2.8Hz, IH), 3.80 (s, 2H).
Figure imgf000068_0003
2-Amino-4-fluoro-6-methyl nitrobenzene: To the crude 3-Amino-5-fluoro-2-nitro phenyl acetic acid (3.6g, 16.8 mmol) was added Q12O (10. lg, 70.6 mmol) in 120 mL of acetonitrile along with 50uL of methanol and the suspension was refluxed for 12 hours. The reaction mixture was filtered through Celite and the Celite pad washed with water and ethyl acetate. The filtrate was extracted with ethyl acetate, washed with water and brine, dried over Na2SO4 and concentrated to give 2.95 g of material which by 1H NMR was 80% pure. ESIMS [M+Na] - 193; 1H NMR (500MHz, DMSO) δ 6.67 (s, 2H), 6.56 (dd, J=l l, 2.8Hz, IH), 6.39 (dd, J=l l, 2.8Hz, IH), 2.50 (s, 3H).
Procedure for the preparation of 4-Chloro-3-(4-methyl-6-moφholin-4-yl-lH- benzoimidazol-2-yl)- 1 H-pyridin-2-one and 4-Iodo-3-(4-methyl-6-moφholin-4-yl- 1 H- benzoimidazol-2-yl)- 1 H-pyridin-2-one
Figure imgf000069_0001
2-Methyl-4-moφholin-4-yl-6-nitro-phenylamine: To a 800 ml pressure flask was added Tris(dibenzylideneacetone)dipalladium (2.64 g, 2.88 mmol), 2-(Di-t- butylphosphino)biphenyl (1.42 g, 4.75 mmol) and sodium tert-butoxide (17.5 g, 182 mmol). Then dry THF (500 mL), 4-bromo-2-methyl-6-nitroaniline (30.0 g, 130 mmol) and moφholine (34 ml, 390 mmol) were added. Argon was bubbled through the solution for 1 minute and the flask was sealed. The reaction mixture was stirred at 85 °C for 3 days. THF was evaporated in vacuo and the crude product was preabsorbed on silica and this then transferred on top of a silica gel column. Elution with hexane-ethyl acetate (6:4 to 4:6 to 0:1 gradient) gave, after evaporation of solvents, the title compound (15.2 g red-brown solid, 49.3%). LCMS (M+H)+ m/z 238 (t = 0.64 min.) 1H NMR (500 MHz, DMSO-d*) δ 7.32 (IH, s), 7.22 (IH, s), 6.96 (2H, s), 3.72 (4H, broad s), 2.96 (4H, broad s), 2.21 (3H, s).
Figure imgf000069_0002
2-(4-Iodo-2-methoxy-pyridin-3-yl)-4-methyl-6-moφholin-4-yl-lH-benzoimidazole: 2-Methyl-4-moφholin-4-yl-6-nitro-phenylamine (15.2 g, 64 mmol) was suspended in methanol (200 ml) in a PARR flask. Palladium on carbon (1.0 g, 10% Pd) was added and the suspension shaken under 60 psi of hydrogen overnight. The mixture was filtered through a pad of celite (under argon) into a 3-neck flask, the celite rinsed with methanol and the filtrate diluted with methanol to a total volume of 500 ml and cooled to 0°C. A solution of 4-Iodo-2-methoxy-pyridine-3-carbaldehyde (14.6 g, 55.5 mmol) in methanol (500 ml) was added slowly (during 3 hours). After addition of - XA of the solution the system was opened to air and stirred over the weekend, thereby reaching room temperature. The reaction mixture was concentrated in vacuo, filtered through a pad of silica (eluent: methylenechloride - ethyl acetate - methanol 55-40-5) then crystallized from ethyl acetate. The title compound was isolated as brown solid (12.68 g, 51%). Flash column chromatography of the mother liquor (gave additional 2.90 g (12%). [pack column with methylene chloride, elute compound with methylene chloride - ethyl acetate 6-4, then methylenechloride - ethyl acetate - methanol 58-40- 2]. LCMS (M+H)+ m/z 451 (t = 1.03 min.). 1H NMR (500 MHz, CDC13) δ 7.76 (IH, d, 7 = 5.3 Hz),), 7.42 (IH, d, 7= 5.3 Hz), 6.85 (IH, broad s), 6.82 (IH, s), 3.86 (4H, t, 7 = 4.5 Hz), 3.79 (3H, s), 3.12 (4H, t, 7 = 4.5 Hz), 2.60 (3H, s), 2.21 (3H, s).
Figure imgf000070_0001
4-Chloro-3-(4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one and 4-Iodo-3-(4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one: 2-(4-Iodo-2-methoxy-pyridin-3-yl)-4-methyl-6-moφholin-4-yl-lH-benzoimidazole (15.58 g, 34.6 mmol) was suspended in 1,4-dioxane (300 ml) and cone, aqueous HCl (50 ml) was added. The mixture was stirred at ambient temperature overnight, then 3 hours at 50°C. The mixture was cooled to room temperature and poured into an ice cold solution of NaHCO3 (67 g, 0.8 mol) and filtered. The aqueous phase was extracted with ethyl acetate. The solid material was dissolved in CH2C12 with some methanol, then extracted between water and CH2CI2 . The combined organic layers were dried over Na2SO4, and concentrated in vacuo to give an inseparable mixture of the 4-Chloro- and 4-Iodo- title compounds. The product was used without further purification. LCMS (M+H)+ m z 437 and m/z 345 (both t = 0.92 min.).
Procedure for the preparation of 4-[2-(4-Chloro-2-oxo-l,2-dihydro-pyridin-3-yl)-7- methyl-3H-benzoimidazol-5-yl]-piperazine-l-carboxylic acid tert-butyl ester and 4- [2-(4-Iodo-2-oxo-l,2-dihydro-pyridin-3-yl)-7-methyl-3H-benzoimidazol-5-yl]- piperazine-1 -carboxylic acid tert-butyl ester
Figure imgf000071_0001
4-(3-Amino-5-methyl-4-nitro-phenyl)-piperazine-l-carboxylic acid tert-butyl ester: To a stirred solution of 3-fluoro-5-amino-6-nitrotoluene (10 g, 58.79 mmol) in anhydrous NMP (160 mL) under nitrogen was addded BOC-piperazine (39 g, 209.4 mmol) and 4-methylmoφholine (25.9 mL). The resulting dark solution was heated to reflux for 72 h, cooled to room temperature and diluted with ethyl acetate (4000 mL). The organic layer was washed with water (8 x 1500 mL), brine (1 x 1500 mL), dried over sodium sulfate and evaporated in vacuo. The resulting dark oil was dissolved in boiling absolute ethanol (800 mL) and concentrated to a total volume of 400 mL and left to stand overnight at room temperature. The solution was further cooled to -20 °C for 5 h and the resulting solid was filtered off and dried in vacuo to give 16.3 g (83%) of a light yellow solid. 1H NMR (500 MHz, CDC13) δ 6.16 (brs, IH), 6.04 (brs, IH), 3.70 - 3.60 (m,4H), 3.38 - 3.25 (m, 4H), 2.53 (s, 3H), 1.48 (s, 9H); LCMS (M+H)+ m/z 337.
Figure imgf000071_0002
4-(3,4-Diamino-5-methyl-phenyl)-piperazine-l -carboxylic acid tert-butyl ester: To a stirred solution of 4-(3-Amino-5-methyl-4-nitro-phenyl)-piperazine-l-carboxylic acid tert-butyl ester (15 g, 44.6 mmol) in methanol (2200 mL) was added 20% Pd(OH)2/C (1.6 g) and the suspension flushed well with nitrogen, followed by hydrogen. The resulting suspension was stirred overnight at room temperature under an atmosphere of hydrogen (ca. 1 atm). The resulting suspension was filtered under nitrogen through a pad of Celite and washed with methanol (400 - 500 mL). The resulting material was used immediately. LCMS (M+H)+ m/z 307.
Figure imgf000072_0001
4-[2-(4-Iodo-2-methoxy-pyridin-3-yl)-7-methyl-3H-benzoimidazol-5-yl]-piperazine- 1 -carboxylic acid tert-butyl ester: To a stirred solution of 4-(3,4-Diamino-5-methyl- phenyl)-piperazine-l -carboxylic acid tert-butyl ester (44.6 mmol - assuming 100% conversion in previous step) in methanol (ca 2700 mL) at 0 oC under a nitrogen atmosphere was slowly added (2 h) via addition funnel, a solution of 4-iodo-2- methoxy-pyridine-3-carbaldehyde (15.0g, 57.1 mmol) in anhydrous methanol (225 mL). The resulting solution was then stirred at 0 °C for an additional 30 min,. the cooling bath was removed, and the reaction mixture was allowed to stir at room temperature in the presence of air for 72 h. The resulting solution was concentrated in vacuo and the residue was dissolved in dichloromethane (1500 mL) and the solvent removed in vacuo (repeated 3x). The resulting dark foamy solid was dried under high vacuum. 1H NMR (500 MHz, CDC13) δ 7.82 (d, IH, J = 5.4 Hz), 7.50 (d, IH, J = 5.4 Hz), 6.98 (brs, IH), 6.90 (brs, IH), 4.05 (s, 3H), 3.67 - 3.58 (m, 4H), 3.18 - 3.09 (m, 4H), 2.63 (s, 3H), 1.49 (s, 9H);. LCMS (M+H)+ m/z 550.
Figure imgf000072_0002
•HCl
4-Iodo-3-(4-methyl-6-piperazin- 1 -yl- 1 H-benzoimidazol-2-yl)- 1 H-pyridin-2-one: To a stirred solution of 4-[2-(4-Iodo-2-methoxy-pyridin-3-yl)-7-methyl-3H-benzoimidazol- 5-yl] -piperazine- 1 -carboxylic acid tert-butyl ester (24g, 43.7 mmol) was added 1,4- dioxane (750 mL) and 6 N aqueous HCl (30 mL) and the mixture was heated to 75 °C overnight. The solution was cooled to room temperature, the supernatent was poured off, and the resulting dark gummy solid was washed with anhydrous diethyl ether (3 x 500 mL) and dried in vacuo to give 17.7 g (93%) of the title compound as a dark solid that was used as described to prepare 4-[2-(4-Chloro-2-oxo-l,2-dihydro-pyridin-3-yl)- 7-methyl-3H-benzoimidazol-5-yl] -piperazine- 1 -carboxylic acid tert-butyl ester and 4- [2-(4-Iodo-2-oxo-l,2-dihydro-pyridin-3-yl)-7-methyl-3H-benzoimidazol-5-yl]- piperazine-1 -carboxylic acid tert-butyl ester; LCMS (M+H)+ m/z 436. (Note: A small peak (4-chloro-pyridin-2-one) in the LC/MS shows m/z 344, 346).
Figure imgf000073_0001
4-[2-(4-Chloro-2-oxo-l,2-dihydro-pyridin-3-yl)-7-methyl-3H-benzoimidazol-5-yl]- piperazine-1 -carboxylic acid tert-butyl ester and 4-[2-(4-Iodo-2-oxo-l,2-dihydro- pyridin-3-yl)-7-methyl-3H-benzoimidazol-5-yl]-piperazine-l-carboxylic acid tert- butyl ester: To a stirred suspension of 4-Iodo-3-(4-methyl-6-piperazin-l-yl-lH- benzoimidazol-2-yl)-lH-pyridin-2-one (17.7 g, 40.7 mmol) in dichloromethane (750 mL) was added di-tert-butyl dicarbonate (9.8 g, 44.8 mmol) and triethylamine (67.4 mL, 483.6 mmol). The mixture was stirred at room temperature for 30 min. and purified via flash chromatography on silica gel. After elution with dichloromethane followed by 2.5% methanol/ethyl acetate, homogeneous fractions were combined and partially evaporated in vacuo to give the product as a yellow solid, following filtration (8.9 g, ca 41%, 2 crops). 1H NMR (500 MHz, CD3OD) δ 7.25 (d, IH, J = 6.9 Hz), 6.97 (d, IH, J = 6.9 Hz), 6.97 (brs, IH), 6.89 (brs, IH), 3.65 - 3.56 (m, 4H), 3.16 - 3.07 (m, 4H), 2.55 (s, 3H), 1.49 (s, 9H); LCMS (M+H)+ m z 536. (Note: A small peak (4-chloro-pyridin-2-one) in the LC/MS shows m/z 444, 446).
Procedure for the preparation of 3-[6-(4-Amino-piperidin-l-yl)-4-methyl-lH- benzoimidazol-2-yl]-4-iodo-lH-pyridin-2-one and 3-[6-(4-Amino-piperidin-l-yl)-4- methyl- 1 H-benzoimidazol-2-yl]-4-chloro- 1 H-pyridin-2-one
Figure imgf000074_0001
[l-(3-Amino-5-methyl-4-nitro-phenyl)-piperidin-4-yl]-carbamic acid tert-butyl ester: 5-Fluoro-3-methyl-2-nitro-phenylamine (0.97 g, 5.7 mmol), 4-N-BOC- aminopiperidine (1.60g, 8.0 mmol), diisopropylethylamine (2.5 ml, 14 mmol) and DMSO (10 ml) are combined and stirred at 85°C for 3 hours. The reaction mixture was poured on saturated aqueous NaHCO3 solution and extracted with ethyl acetate. The organic layers were washed with water (3x) and brine, dried over Na2SO4 and concentrated. Rash column chromatography on silica (eluent hexanes-ethyl acetate - triethylamine 50-50-1, then 33-66-1) gave the title compound as a yellow solid. (1.57 g, 79%). LCMS (M+H)+ m/z 351 (t = 1.55 min.). 1H NMR (500 MHz, CD3OD) δ 6.70 (IH, broad s), 6.22 (IH, d, 7 = 2.5 Hz)), 6.13 (IH, d, 7= 2.5 Hz), 3.88 (2H, d, 7 = 13.3 Hz), 3.58 (IH, broad s), 2.98 (2H, t, 7 = 11.8 Hz), 2.48 (3H, s), 1.92 (2H, d, 7 = 11.3 Hz), 1.48 (2H, m), 1.45 (9H, s).
Figure imgf000074_0002
{ l-[2-(4-Iodo-2-methoxy-pyridin-3-yl)-7-methyl-3H-benzoimidazol-5-yl]-piperidin- 4-yl}-carbamic acid tert-butyl ester: [l-(3-Amino-5-methyl-4-nitro-phenyl)- piperidin-4-yl]-carbamic acid tert-butyl ester (1.54 g, 4.4 mmol) was dissolved in methanol (100 ml). Palladium on carbon (0.3 g, 10% Pd) was added and the suspension stirred vigorously under a balloon pressure of hydrogen overnight. The mixture was filtered through a pad of celite (under argon) into a 3-neck flask, the celite rinsed with methanol and the filtrate cooled to 0°C. A solution of 4-Iodo-2- methoxy-pyridine-3-carbaldehyde (1.21 g, 4.6 mmol) in methanol (50 ml) was added slowly (during 2 hours). The mixture was stirred overnight under air at ambient temperature, then concentrated in vacuo. Rash column chromatography on silica (eluent hexanes-ethyl acetate -methanol 5-4-1 gave the title compound. (0.79 g, 32%). LCMS (M+H)+ m/z 564 (t = 1.31 min.).
Figure imgf000075_0001
3- [6-(4- Amino-piperidin- 1 -yl)-4-methyl- 1 H-benzoimidazol-2-yl]-4-iodo- 1 H-pyridin- 2-one and 3 -[6-(4- Amino-piperidin- l-yl)-4-methyl-lH-benzoimidazol-2-yl]-4-chloro- lH-pyridin-2-one : { l-[2-(4-Iodo-2-methoxy-pyridin-3-yl)-7-methyl-3H- benzoimidazol-5-yl]-piperidin-4-yl}-carbamic acid tert-butyl ester (330 mg, 0.59 mmol) was suspended in 4M HCl in 1,4-dioxane (20 ml) and water (3 ml) was added, (exothermic reaction). The mixture was stirred at ambient temperature overnight, then concentrated in vacuo to give an inseparable mixture of the 4-Chloro- and 4-Iodo- title compounds. The product was used without further purification. LCMS (M+H)+ m/z 450 and m/z 358 (both t = 0.69 min.).
Figure imgf000075_0002
3-Methyl-5-(2-moφholin-4-ethoxy)-2-nitro-phenylamine: To a solution of 2- moφholin-4-yl-ethanol (5 g, excess) in THF (30 mL) was added NaH (0.21 g, 8.82 mmol) in portion under ice bath. The reaction mixture was stirred at room temperature for 30 min. Then 5-fluoro-3-methyl-2-nitro-phenylamine was added. The reaction mixture was heated to reflux for 6 h, cooled to room temperature, and concentrated. The residue was diluted with water and extracted with EtOAc. The combined organic layers were washed with water, brine, and dried over Na2SO . After concentration, the residue was purified by column chromatography (20% EtOAc / hexane) to yield the title compound (0.70 g, 85%). 1H NMR (400 MHz, CD3OD) δ 6.10 (IH, s), 6.09 (IH, s), 4.38 - 4.42 (2H, m), 3.92 - 4.08 (4H, m), 3.72 (IH, d, 7= 12 Hz), 3.53 - 3.56 (2H, m), 3.05 - 3.10 (2H, m), 2.48 (3H, s). LCMS (M+H)+ m/z 282 (t = 0.73 min.).
Figure imgf000076_0001
(S)-7-Bromo-2-{4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl}-3H-benzimidazole-5-carbaldehyde: To a solution of (S)-7-Bromo-2- { 4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro-pyridin-3-yl } -3H- benzimidazole-5-carbonitrile (150 mg, 0.31 mmol) in THF (50 mL) was added DIBAL-H (1 M toluene solution, 1.55 mL, 1.55 mmol) at -78°C. The reaction mixture was stirred at -40°C for 10 h and cooled to -78°C. EtOAc (0.5 mL) was added. The reaction mixture was stirred for 30 min at-78°C before water (1 mL) was added. The reaction mixture was warmed to room temperature and concentrated. The residue was passed through a small pad of celite. The filtrate was concentrated and purified by prep. HPLC to give the titled compound (67 mg, 43%). 1H NMR (400 MHz, CD3OD) δ 9.45 (IH, s), 7.62 (IH, s), 7.56 (IH, narrow d, 7 = 1.6 Hz), 7.44 (IH, narrow d, 7= 1.0 Hz), 7.32 - 7.42 (2H, m), 7.24 - 7.30 (3H, m), 6.24 (IH, d, 7= 7.6 Hz), 5.01 (IH, m), 3.65 - 3.76 (2H, m). LCMS (M+H)+ m/z 487 (t = 1.76 min.).
Figure imgf000076_0002
5-(l,4,5,6-Tetrahydropyrimidin-l-yl)-3-methyl-2-nitro aniline: To a stirred solution of 2.0 g (11.76 mmol) of the 5-Flouro-3-methyl-2-nitro aniline in 10 mL of DMSO was added 1.2 g (14.11 mmol) of 1,4,5,6-Tetrahydropyrimidine, and 2.43 g (17.64 mmol) of potassium carbonate, and the mixture was heated at 100°C for 10 hrs, cooled, diluted with water, and extracted with Ethylacetate containing 5% methanol. The combined organic extract was washed with water, brine and dried (Na2SO4). Evaporation of the solvent furnished the residue, which was chromatographed (20% of 2M ammonia in methanol and dichloromethane) to produce 1.85 g (67%) of the product as red solid. 1H NMR (400 MHz, CD3OD) δ 7.89 (lH,s), 6.53 (lH,d, 7= 2.57 Hz), 6.44 ((lH,d, 7= 2.1 Hz), 7.04 (IH, d, 7 = 2.1Hz), 3.70 (2H, t, 7= 6.0 Hz), 3.41 (2H, t, J = 5.65 Hz, 2.43 (3H, s), 2.05 (2H, m) LCMS (M+H)+ m/z 235 (t = 0.78 min).
Figure imgf000077_0001
(S)-2-Amino-l-[3-cynaophenyl]ethanol Hydrochloride: was prepared according to the general procedure outlined above. LRMS [M+H]+ 163; IH NMR (500MHz) δ 7.64 (brs, IH), 7.47 (d, IH), 7.37 (d, IH), 7.25 (t, IH), 4.85 (dd, IH), 3.05 (dd, IH), 2.75 (dd, IH).
Figure imgf000077_0002
4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[6-(4-isocyanato-piperidin- 1 -yl)-4- methyl-lH-benzoimidazol-2-yl]-lH-pyridin-2-one : To a solution of the 4-amino- piperidine compound (1.5g) in THF (15ml) was added Di-t-butyltricarbonate [Dean, C. S. et al. 7. Org. Chem., 35; 1970; 3393-3397] (1.04g, 1.3 eq) in THF (15ml) at room temperature. The reaction mixture was stirred at r.t. for lhr and concentrated and was used as it is. LCMS (M+H)+ m/z 519 (t = 1.84 min, YMC Xterra C18 S7 3.0 x 50 mm; 0 - 100% gradient over 2 min; 5 mL/min flow rate).
Figure imgf000078_0001
( 1 -Tert-butyl oxycarbonyl-4-methyl-6-moφholin-4-yl- lH-benzoimidazol-2-yl)-acetic acid ethyl ester: A solution of 2-methyl-4-moφholin-4-yl-6-nitro-phenylamine (1.84 g, 7.75 mmol) in methanol (115 mL) was hydrogenated (15 psi) in presence of palladium(II) hydroxide for 12 hours. The reaction was filtered and the filtrate was evaporated. The crude amine was dissolved in N,N-dimethylformamide (70 mL) and treated with the hydrochloride salt of ethyl mono-imido malonate (3.04 g, 15.48 mmol) and this mixture was stirred at 70°C for 2 hours. The solvent was evaporated and the residue was dissolved in dichloromethane (100 mL) and treated with triethylamine (2.16 mL, 15.48 mmol), di-tert-butyl dicarbonate (3.38 g, 15.48 mmol) and dimethylaminopyridine (2 crystals). The reaction was stirred at RT for 1.5 hours, then saturated ammonium chloride was added and the mixture was extracted with ethyl acetate (3x). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (50% ethyl acetate in hexane) to give the title material (1.955 g, 63%) as a brown solid. LCMS (+ESI, M+Η+) m/z 404; 1H NMR (400 MHz, DMSO-de) δ (ppm): 1.26 (3H, t, J=7.1 Hz), 1.70 (9H, s), 2.56 (3H, s), 3.21 (4H, br dd), 3.85 (4H, br dd), 4.18 (2H, qa, J=7.1 Hz), 4.27 (2H, s), 7.02 (IH, d, J-1.5 Hz), 7.33 (lH, d, J=2.0 Hz).
Figure imgf000078_0002
[2-( 1 -Tert-butyl oxycarbonyl-4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- acetylamino] -acetic acid ethyl ester: To a stirred solution of (1-tert-butyloxycarbonyl- 4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-acetic acid ethyl ester (0.400 g, 0.99 mmol) in ethanol (15 mL) at 0°C was added aqueous sodium hydroxide (5N, 1 mL). The mixture was stirred at RT for ~1 hour, then cooled down to 0°C and neutralized with aqueous hydrochloric acid (IN) until the pΗ reaches ~7. Ethyl acetate was added and the aqueous phase was washed with ethyl acetate (3x). The aqueous phase was evaporated to remove the last traces of ethanol and was then lyophilized.
The resulting solid (0.499 g) was suspended in N,N-dimethylformamide (15 mL) and treated with triethylamine (0.553 mL, 3.96 mmol), the hydrochloric acid salt of glycine ethyl ester (0.207 g, 1.49 mmol) and benzotriazole-1-yl-oxy-tris-pyrrolidino- phosphonium hexafluorophosphate (PyBOP) (1.03 g, 1.98 mmol). The mixture was stirred at RT for 5 minutes then the solvent was evaporated. The residue was dissolved in ethyl acetate and washed with water (3x). The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was dissolved in dichloromethane (12 mL) and treated with triethylamine (0.250 mL), di-tert-butyl dicarbonate (0.325 g, 1.49 mmol) and 2,4-dimethylamino pyridine (1 crystal). The reaction was stirred at RT during the week-end. Saturated ammonium chloride solution was added and the aqueous phase was extracted with dichloromethane (3x). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by chromatography (ethyl acetate) to give the title material (0.481 g, 83%). ΗPLC: 89% (220 nm), LCMS (+ESI, M+Η+) m z 461; 1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.17 (3H, t, J=7.2 Hz), 1.58 (9H, s), 2.45 (3H, s), 3.09 (4H, br dd), 3.75 (4H, br dd), 3.83 (2H, d, J=5.8 Hz), 4.07 (2H, qa, J=7.2 Hz), 4.09 (2H, br s), 6.88 (IH, br dd), 7.23 (IH, d J=2.3 Hz), 8.50 (IH, t, J=5.8 Hz).
Figure imgf000079_0001
4-Hydroxy-3-(l-tert-butyloxycarbonyl-4-methyl-6-moφholin-4-yl-lH- benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one: To a stirred suspension of sodium hydride (60% in oil, 0.014 g, 0.352 mmol)in toluene (5 mL) was added [2-(l-tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-acetylamino]- acetic acid ethyl ester (0.135 g, 0.293 mmol) in tetrahydrofuran (5 mL) over 5 minutes. The mixture was stirred at 90-100°C for 2.5 hours, then cooled down to ~0°C and ethyl acetate followed by aqueous ammonium chloride (~3 mL) were added. The aqueous phase was extracted with ethyl acetate (3x). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was quickly purified by silica gel chromatography to give the title material (0.072 g, 59%) as a white solid. LCMS (+ESI, M+H+) m/z 415, (" ESI, M-H") m/z 413; 1H NMR (400 MHz, DMSO-d^) δ (ppm): 1.47 (9H, s), 3.06 (4H, br s), 3.74 (4H, br s), 3.94 (2H, br s), 6.83 (IH, s), 6.97 (IH, s), 12.1 (IH, m), 12.56 (lH, s).
Figure imgf000080_0001
4-Hydroxy-3-(4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5-dihydro- pyrrol-2-one: To a stirred suspension of sodium hydride (60% in oil, 0.003 g, 0.044 mmol) in toluene (1 mL) was added [2-(l-tert-butyloxycarbonyl-4-methyl-6- moφholin-4-yl-lH-benzoimidazol-2-yl)-acetylamino] -acetic acid ethyl ester (0.135 g, 0.293 mmol) in tetrahydrofuran (2 mL) over 5 minutes. The mixture was stirred at 90-100°C for 5 hours, then cooled down to ~0°C and ethyl acetate followed by aqueous hydrochloric acid (IN, -0.5 mL) were added. The aqueous phase was washed with ethyl acetate (3x) and this was submitted to Prep HPLC (acetonitrile/ammonium acetate/water) to give the title material (0.008 g, 69%) as a white solid. HPLC 100% (220 nm); LCMS (+ESI, M+H+) m/z 315; 1H NMR (400 MHz, methanol- ) δ (ppm): 2.55 (3H, s), 3.18 (4H, br dd), 3.81 (2H, s), 3.88 (4H, br dd), 6.92 (IH, br s), 6.99 (IH, d, J=2.0 Hz).
Figure imgf000081_0001
(S)-2-[2-(l-tert-butyloxycarbonyl-4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2- yl)-acetylamino]-propionic acid ethyl ester: (4-Methyl-6-moφholin-4-yl-lH- benzoimidazol-2-yl)-acetic acid (0.250 g, -0.574 mmol, prepared as described in the procedure to synthesize [2-(l-tert-butyloxycarbonyl-4-methyl-6-moφholin-4-yl-lΗ- benzoimidazol-2-yl)-acetylamino]-acetic acid ethyl ester) was reacted as described in the same procedure using L-alanine ethyl ester instead of glycine ethyl ester to give the title material (0.197 g, 72%). LCMS (+ESI, M+lT") m z 475; 1H NMR (400 MHz, DMSO-de) δ (ppm): 1.16 (3H, t, J=7.1 Hz), 1.28 (3H, d, J=7.3 Hz), 1.58 (9H, s), 2.44 (3H, s), 3.09 (4H, br dd), 3.75 (4H, br dd), 4.02 (2H, qa, J=7.1 Hz), 4.05 (2H, m), 4.23 (IH, m), 6.88 (IH, br s), 7.23 (IH, d, J=2.0 Hz), 8.57 (IH, d, J=6.8 Hz).
Figure imgf000081_0002
(S and R)-4-Hydroxy-5-methyl-3-(l -tert-butyloxycarbonyl-4-methyl-6-moφholin-4- yl- 1 H-benzoimidazol-2-yl)- 1 ,5-dihydro-pyrrol-2-one: (S)-2-[2-(l -tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-acetylamino]- propionic acid ethyl ester (0.195 g, 0.411 mmol) was reacted as described in the procedure used to synthesize 4-hydroxy-3-(l-tert-butyloxycarbonyl-4-methyl-6- moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5-dihydro-ρyrrol-2-one and gave the title material (0.03 lg, 18% precipitated from ethyl acetate and 0.129 g crude, 73%). HPLC 91% (220 nm), LCMS (+ESI, M+H+) m/z 429; *H NMR (400 MHz, methanol-cL δ (ppm): 1.50 (3H, d, J=6.9 Hz), 1.57 (9H, s), 2.54 (3H, s)3.16 (4H, br dd), 3.85 (4H, br dd), 4.17 (IH, m), 6.15 (IH, s), 6.93 (IH, s).
Example 1 (General procedure for examples 1-21)
Figure imgf000082_0001
(S)-4-(2-Hydroxy- 1 -phenyl -ethylamino)-3-(6-imidazol- 1 -yl-4-methyl- 1 H- benzimidazol-2-yl)-lH-pyridin-2-one: To a solution of 3-(6-imidazol-l-yl-4-methyl- lH-benzimidazol-2-yl)-4-iodo-lH-pyridin-2-one (30 mg, 0.072 mmol) in DMF (1 mL) were added (S)-(-)-2-phenylglycinol (26 mg, 0.18 mmol) and N- methylmoφholine (0.1 mL). The reaction mixture was heated to 80°C for 6 h and cooled to room temperature. The solvent was removed under vacuum and the residue was purified by prep. HPLC to yield the title compound (16 mg, 52%). 1H ΝMR (300 MHz, CD3OD) δ 8.07 (IH, narrow t, 7= 1.7 Hz), 7.76 (2H, s), 7.27 - 7.48 (7H, m), 7.21 (IH, d, 7 = 7.5 Hz), 6.11 (IH, d, 7 = 7.5 Hz), 4.92 (IH, m), 4.03 (IH, dd, 7 = 4.5, 11.2 Hz), 3.95 (IH, dd, 7 = 6.2, 11.2 Hz), 2.75 (3H, s). LCMS (M+H)+ m/z 427 (t = 1.44 min.)
Example 2
Figure imgf000082_0002
(±)-4-[2-Hydroxy-2-(3-iodo-phenyl)-ethylamino]-3-(6-imidazol-l-yl-4-methyl-lH- benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.41 (IH, s), 8.06 (IH, s), 7.88 (IH, s), 7.76 (IH, s), 7.71 (IH, s), 7.59 (IH, d, 7= 7.8 Hz), 7.47 (IH, d, 7 = 7.8 Hz) 7.32 (IH, s) 7.29 (IH, d, 7 = 7.6 Hz) 7.09 (IH, t, 7 = 7.8 Hz), (6.24 (IH, d, 7 = 7.6 Hz), 4.97 (IH, dd, 7 = 5.0, 6.0 Hz), 3.75 (IH, dd, 7 = 5.0, 13.5 Hz), 3.67 (IH, dd, 7= 6.0, 13.5 Hz), 2.68 (3H, s). LCMS (M+H)+ m/z 553 (t = 1.43 min.).
Example 3
Figure imgf000083_0001
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(6-imidazol-l-yl-4-methyl-lH- benzimidazol-2-yl)-lH-pyridin-2-one: *H NMR (300 MHz, CD3OD) δ 9.39 (IH, narrow t, 7 = 1.4 Hz), 8.04 (IH, narrow t, 7= 1.7 Hz), 7.76 (IH, narrow t, 7 = 1.7 Hz), 7.69 (IH, narrow d, 7 = 1.9 Hz), 7.55 (IH, s), 7.23 - 7.42 (5H, m), 6.25 (IH, d, 7 = 7.6 Hz), 5.01 (IH, dd, 7 = 4.8, 6.4 Hz), 3.76 (IH, dd, 7 = 4.8, 13.4 Hz), 3.66 (IH, dd, 7 = 6.4, 13.4 Hz), 2.67 (3H, s). LCMS (M+H)+ m/z 461 (t = 1.46 min.).
Example 4
Figure imgf000083_0002
(±)-4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-(6-imidazol-l-yl-4-methyl-lH- benzimidazol-2-yl)-lH-pyridin-2-one: Η NMR (300 MHz, CD3OD) δ 9.38 (IH, s), 8.02 (IH, s), 7.74 (IH, s), 7.69 (IH, s), 7.66 (IH, narrow d, 7= 1.4 Hz), 7.20 - 7.48 (5H, m), 6.21 (IH, d, 7 = 7.6 Hz), 4.99 (IH, dd, 7 = 4.8, 6.3 Hz), 3.73 (IH, dd, 7 = 4.8, 13.5 Hz), 3.64 (IH, dd, 7 = 6.3, 13.5 Hz), 2.65 (3H, s). LCMS (M+H)+ m/z 505 (t = 1.47 min.).
Example 5
Figure imgf000084_0001
(±)-N-(2-Chloro-4- { 1 -hydroxy-2- [3-(6-imidazol- 1 -yl-4-methyl- lH-benzimidazol-2- yl)-2-oxo- 1 ,2-dihydro-pyridin-4-ylamino]-ethyl } -phenyl)-methanesulfonamide: 1H ΝMR (300 MHz, CD3OD) δ 9.39 (IH, s), 8.05 (IH, s), 7.76 (IH, s), 7.68 (IH, s), 7.62 (IH, narrow d, 7 = 1.5 Hz), 7.52 (IH, s), 7.42 - 7.49 (2H, m), 7.31 (IH, s), 7.30 (IH, d, 7 = 7.6 Hz), 6.26 (IH, d, 7 = 7.6 Hz), 5.01 (IH, dd, 7 = 5.0, 5.6 Hz), 3.76 (IH, d, 7 = 5.0, 13.4 Hz), 3.74 (IH, dd, 7= 5.6, 13.4 Hz), 2.98 (3H, s), 2.68 (3H, s). LCMS (M+H)+ m z 554 (t = 1.11 min.). .
Example 6
Figure imgf000084_0002
3-Bromo-benzoic-acid N-[3-(6-imidazol-l-yl-4-methyl-lH-benzimidazol-2-yl)-2- oxo-l,2-dihydro-pyridin-4-yl]-hydrazide: 1H NMR (300 MHz, CD3OD) δ 9.40 (IH, narrow t, 7 = 1.4 Hz), 8.14 (IH, narrow t, 7 = 1.7 Hz), 8.05 (IH, narrow t, 7 = 1.7 Hz), 7.95 (IH, d, 7 = 7.9 Hz), 7.81 (IH, d, 7 = 7.9 Hz), 7.74 - 7.77 (2H, m), 7.49 (IH, t, 7 = 7.9 Hz), 7.41 (IH, d, 7 = 7.4 Hz), 7.34 (IH, s), 6.38 (IH, d, 7 = 7.4 Hz), 2.70 (3H, s). LCMS (M+H)+ m/z 504 (t = 1.44 min.).
Example 7
Figure imgf000085_0001
4-Amino-benzoic-acid ΛT -[3-(6-imidazol- 1 -yl-4-methyl- lH-benzimidazol-2-yl)-2- oxo-l,2-dihydro-pyridin-4-yl]-hydrazide: 1H NMR (300 MHz, CD3OD) δ 9.41 (IH, s), 8.06 (IH, s), 7.76 - 7.79 (4H, m), 7.39 (IH, d, 7= 7.4 Hz), 7.36 (IH, s), 6.81 (2H, d, 7 = 8.6 Hz), 6.36 (IH, d, 7 = 7.4 Hz), 2.70 (3H, s). LCMS (M+H)+ m/z 441 (t = 0.96 min.).
Example 8
Figure imgf000085_0002
(S)-4-[2-(2-Chloro-phenyl)- 1 -hydroxymethyl-ethylamino]-3-(6-imidazol- 1 -yl-4- methyl-lH-benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.41 (IH, s), 8.05 (IH, narrow t, 7= 1.7 Hz), 7.76 (IH, narrow t, 7= 1.7 Hz), 7.72 (IH, narrow d, 7= 1.9 Hz), 7.08 - 7.36 (6H, m), 6.10 (IH, d, 7= 7.7 Hz), 3.98 - 4.24 (IH, m), 3.84 (IH, dd, 7= 4.4, 11.2 Hz), 3.79 (IH, dd, 7= 4.8, 11.2 Hz), 3.35 (IH, dd, 7 = 5.4, 13.6 Hz), 3.09 (IH, dd, 7= 7.8, 13.6 Hz), 2.72 (3H, s). LCMS (M+H)+ m/z 475 (t = 1.56 min.).
Example 9
Figure imgf000086_0001
(S)-4-[2-(3-Chloro-phenyl)-l-hydroxymethyl-ethylamino]-3-(6-imidazol-l-yl-4- methyl-lH-benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.42 (IH, s), 8.07 (IH, narrow t, 7= 1.7 Hz), 7.75 - 7.78 (2H, m), 7.14 - 7.37 (6H, m), 6.18 (IH, d, 7 = 7.7 Hz), 4.07 - 4.11 (IH, m), 3.76 - 3.77 (2H, m), 3.17 (IH, dd, 7 = 5.1, 13.7 Hz), 2.98 (IH, dd, 7= 8.2, 13.7 Hz), 2.71 (3H, s). LCMS (M+H)+ m/z 475 (t = 1.57 min.).
Example 10
Figure imgf000086_0002
(S)-4-[2-(4-Chloro-phenyl)- 1 -hydroxymethyl-ethylamino]-3-(6-imidazol- 1 -yl-4- methyl-lH-benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.42 (IH, s), 8.07 (IH, narrow t, 7= 1.6 Hz), 7.77 (IH, narrow t, 7 = 1.6 Hz), 7.73 (IH, narrow d, 7 = 1.9 Hz), 7.16 - 7.37 (6H, m), 6.19 (IH, d, 7= 7.7 Hz), 4.06 - 4.10 (IH, m), 3.72 - 3.77 (2H, m), 3.14 (IH, dd, 7 = 5.3, 13.8 Hz), 2.98 (IH, dd, 7 = 7.8, 13.8 Hz), 2.69 (3H, s). LCMS (M+H)+ m/z 475 (t = 1.61 min.). Example 11
Figure imgf000087_0001
(S)-4-[2-(2-Bromo-phenyl)- 1 -hydroxymethyl-ethylamino]-3-(6-imidazol- 1 -yl-4- methyl-lH-benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.41 (IH, s), 8.06 (IH, s), 7.75 - 7.77 (2H, m), 7.52 (IH, dd, 7 = 1.5, 7.5 Hz), 7.36 (IH, d, 7= 1.9 Hz), 7.34 (IH, s), 7.03 - 7.16 (3H, m), 6.09 (IH, d, 7 = 7.7 Hz), 4.15 4.27 (IH, m), 3.82 (2H, m), 3.35 (IH, dd, 7 = 5.0, 13.6 Hz), 3.10 (IH, dd, 7= 9.0, 13.6 Hz), 2.74 (3H, s). LCMS (M+H)+ m/z 519 (t = 1.56 min.).
Example 12
Figure imgf000087_0002
(S)-4-[2-(3-Bromo-phenyl)-l-hydroxymethyl-ethylamino]-3-(6-imidazol-l-yl-4- methyl-lH-benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.41 (IH, s), 8.06 (IH, s), 7.74 - 7.77 (2H, m), 7.47 (IH, s), 7.24 - 7.31 (4H, m), 7.11 (IH, d, 7= 7.7 Hz), 6.16 (IH, d, 7 = 7.7 Hz), 4.05 - 4.11 (IH, m), 3.76 (2H, m), 3.15 (IH, dd, 7= 5.0, 13.6 Hz), 2.96 (IH, dd, 7 = 8.3, 13.6 Hz), 2.70 (3H, s). LCMS (M+H)+ m/z 519 (t = 1.54 min.). Example 13
Figure imgf000088_0001
(+)-4-(l-Hydroxymethyl-2-pentafluorophenyl-ethylamino)-3-(6-imidazol-l-yl-4- methyl-lH-benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.40 (IH, s), 8.06 (IH, narrow t, 7 = 1.8 Hz), 7.77 (IH, narrow t, 7 = 1.7 Hz), 7.74 (IH, narrow d, 7 = 1.8 Hz), 7.35 (IH, s), 7.29 (IH, d, 7 = 7.6 Hz), 6.22 (IH, d, 7 = 7.6 Hz), 4.24 (IH, m), 3.82 (2H, dd, J = 2.6, 4.5 Hz), 3.23 (2H, t, 7 = 6.5 Hz), 2.70 (3H, s). LCMS (M+H)+ m/z 531 (t = 1.61 min.).
Example 14
Figure imgf000088_0002
(S)-4-( 1 -Hydroxymethyl-2-pyridin-4-yl-ethylamino)-3-(6-imidazol- 1 -yl-4-methyl- 1 H- benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 9.42 (IH, s), 8.67 (2H, d, 7= 6.6 Hz), 8.07 (2H, d, 7 = 6.6 Hz), 8.06 (IH, s), 7.77 (2H, s), 7.36 (IH, s), 7.28 (IH, d, 7 = 7. 6 Hz), 6.24 (IH, d, 7 = 7.6 Hz), 4.35 (IH, m), 3.82 (2H, d, 7 = 4.4 Hz), 3.50 (IH, dd, 7= 4.4, 13.6 Hz), 3.40 (IH, dd, 7= 8.7, 13.6 Hz), 2.71 (3H, s). LCMS (M+H)+ m/z 442 (t = 0.96 min.). Example 15
Figure imgf000089_0001
(S)-4-( 1 -Hydroxymethyl-2-naphthalen-2-yl-ethylamino)-3-(6-imidazol- 1 -yl-4-methyl- lH-benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.99 (IH, narrow t, 7 = 1.7 Hz), 7.26 - 7.75 (11H, m), 7.15 (IH, d, 7 = 7.6 Hz), 6.19 (IH, d, 7 = 7.6 Hz), 4.16 - 4.20 (IH, m), 3.75 - 3.86 (2H, m), 3.30 (IH, dd, 7 = 5.4, 13.6 Hz), 3.15 (IH, dd, 7= 7.6, 13.6 Hz), 2.60 (3H, s). LCMS (M+H)+ m/z 491 (t = 1.71 min.).
Example 16
Figure imgf000089_0002
(S)-4-(2-Cyclohexyl- 1 -hydroxymethyl-ethylamino)-3-(6-imidazol- 1 -yl-4-methyl- 1 H- benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.40 (IH, s), 8.06 (IH, narrow t, 7 = 1.7 Hz), 7.76 (IH, narrow t, 7 = 1.7 Hz), 7.73 (IH, narrow d, 7 = 1.7 Hz), 7.34 (IH, d, 7= 7.6 Hz), 7.33 (IH, s), 6.34 (IH, d, 7 = 7.6 Hz), 3.89 - 3.94 (IH, m), 3.68 (2H, d, 7 = 4.9 Hz), 2.68 (3H, s), 1.62 - 1.83 (7H, m), 0.95 - 1.26 (6H, m). LCMS (M+H)+ m/z 447 (t = 1.71 min.). Example 17
Figure imgf000090_0001
(3S,4R)-4-(3-Hydroxy-2,2-dimethyl-chroman-4-ylamino)-3-(6-imidazol-l-yl-4- methyl-lH-benzimidazol-2-yl)-lH-pyridin-2-one: *H NMR (300 MHz, CD3OD) δ 9.36 (IH, s), 8.02 (IH, narrow t, 7 = 1.7 Hz), 7.73 (IH, narrow t, 7 = 1.7 Hz), 7.69 (IH, narrow d, 7 = 2.0 Hz), 7.37 (IH, d, 7= 7.6 Hz), 7.18 - 7.30 (3H, m), 6.89 (IH, t, 7= 7.6 Hz), 6.85 (IH, d, 7= 8.4 Hz), 6.61 (IH, d, 7= 7.6 Hz), 4.93 (IH, d, 7 = 8.2 Hz), 3.80 (IH, d, 7= 8.2 Hz), 2.46 (3H, s), 1.48 (3H, s), 1.35 (3H, s). LCMS (M+H)+ m/z 483 (t = 1.70 min.).
Example 18
Figure imgf000090_0002
3-(6-Imidazol-l-yl-4-methyl-lH-benzimidazol-2-yl)-4-(2-thiophen-2-yl-ethylamino)- lH-pyridin-2-one: 1H NMR (500 MHz, DMSO-de) δ 13.17 (IH, broad s), 11.32 (IH, broad s), 11.03 (IH, broad s), 9.59 (IH, broad s), 8.22 (IH, broad s), 7.91 (IH, s), 7.82 (IH, d, 7 = 2.0 Hz), 7.41 (IH, t, 7= 6.9 Hz), 7.36 (IH, dd, 7 = 1.0, 5.1 Hz), 7.30 - 7.40 (IH, m), 7.05 (IH, broad s), 7.01 (IH, t, 7 = 4.2 Hz), 6.23 (IH, d, 7 = 7.5 Hz), 3.74 (2H, t, 7 = 6.5 Hz), 3.25 (2H, t, 7 = 6.5 Hz), 2.58 (3H, s). Example 19
Figure imgf000091_0001
3-(6-Imidazol- 1 -yl-4-methyl- 1 H-benzimidazol-2-yl)-4-[2-(l H-indol-3-yl)- ethylamino]-lH-pyridin-2-one: 1H NMR (500 MHz, DMSO-de) δ 13.17 (IH, broad s), 11.27 (broad s, IH), 11.00 (IH, broad s), 10.89 (IH, s), 9.58 (IH, broad s), 8.30 (IH, broad s), 7.91 (IH, s), 7.80 (IH, s), 7.63 (IH, d, 7= 7.9 Hz), 7.40 (IH, t, 7= 6.9 Hz), 7.34 (IH, d, 7 = 8.0 Hz), 7.30 - 7.40 (2H, m), 7.07 (IH, t, 7 = 7.5 Hz), 6.99 (IH, t, 7= 7.6 Hz), 6.23 (IH, dd, 7 = 0.8, 7.2 Hz), 3.76 (2H, broad s), 3.17 (2H, t, 7= 6.7 Hz), 2.50 (3H, s).
Example 20
Figure imgf000091_0002
3-(6-Irnidazol-l-yl-4-methyl-lH-benzimidazol-2-yl)-4-(pyridin-2-ylmethoxy)-lH- pyridin-2-one: To a solution of 3-(6-imidazol-l-yl-4-methyl-lH-benzimidazol-2-yl)- 4-iodo-lH-pyridin-2-one (25 mg, 0.06 mmol) in DMF (2 mL) were added pyridine carbinol (26 mg, 0.24 mmol) and cesium fluoride (36 mg, 0.24 mmol). The reaction mixture was heated to 130°C for 14 h and cooled to room temperature. After concentration in vacuo, the residue was purified by prep. HPLC to yield the title compound (8.2 mg, 34%). *H NMR (300 MHz, CD3OD) δ 9.58 (IH, narrow d, 7 = 1.2 Hz), 8.92 (IH, d, 7 = 4.9 Hz), 8.19 (IH, narrow t, 7 = 1.2 Hz), 8.12 (IH, s), 7.97 - 8.02 (2H, m), 7.85 (IH, narrow t, 7 = 1.8 Hz), 7.81 (IH, narrow t, 7 = 1.0 Hz), 7.63 (IH, d, 7= 7.9 Hz), 7.54 (IH, t, 7= 6.2 Hz), 6.81 (IH, 7 = 7.5 Hz), 5.86 (2H, s), 2.86 (3H, s). LCMS (M+H)+ m z 399 (t = 1.07 min.).
Example 21
Figure imgf000092_0001
(±)-4-[2-(3-Bromo-phenyl)-2-fluoro-ethylamino]-3-(6-imidazol- 1 -yl-4-methyl- 1H- benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.40 (IH, t, 7 = 1.4 Hz), 8.06 (IH, t, 7= 1.9 Hz), 7.76 (IH, t, 7 = 1.7 Hz), 7.72 (IH, d, 7= 1.9 Hz), 7.66 (IH, s), 7.27 - 7.50 (5H, m), 6.29 (IH, d, 7 = 7.6 Hz), 5.75 - 5.94 (IH, m), 3.86 - 4.06 (2H, m), 2.64 (3H, s). LCMS (M+H)+ m z 507 (t = 1.70 min.).
Example 22 (General procedure for examples 22-28)
Figure imgf000092_0002
(S)-2-[4-( 1 -Hydroxymethyl-2-phenyl-ethylamino)-2-oxo- 1 ,2-dihydro-pyridin-3-yl] -7- methyl-3H-benzimidazole-5-carbonitrile: To a solution of 2-(4-chloro-2-oxo-l,2- dihydro-pyridin-3-yl)-7-methyl-3H-benzimidazole-5-carbonitrile (0.7 g, 2.19 mmol) in DMF (15 mL) were added N-methylmoφholine (0.66 g, 6.57 mmol) and (S)-(-)-2- amino-3-phenyl-l-propanol (0.40 g, 2.63 mmol). The reaction mixture was heated to 80°C for 6 h and then cooled to room temperature. After concentration in vacuo, the residue was purified by flash chromatography (3% MeOH / CH2CI2) to yield the title compound (0.59 g, 68%) as a yellow foam.1H ΝMR (400 MHz, CD3OD) δ 7.74 (IH, s), 7.63 (IH, s), 7.12 - 7.27 (6H, m), 6.07 (IH, d, 7= 7.5 Hz), 3.97 (IH, m), 3.74 (2H, t, 7= 5 Hz), 3.14 (IH, dd, 7 = 5.5, 14.0 Hz), 2.94 (IH, dd, 7= 7.9, 14.0 Hz), 2.60 (3H, s). LCMS (M+H)+ m/z 400 (t = 1.71 min.).
Example 23
Figure imgf000093_0001
(±)-2- { 4- [2-(3 -Chloro-phenyl)-2-hydroxy-ethylamino] -2-oxo- 1 ,2-dihydro-pyridin-3 - yl}-7-methyl-3H-benzimidazole-5-carbonitrile: 1H NMR (300 MHz, DMSO-de) δ 7.92 (IH, s), 7.59 (IH, s), 7.28 - 7.47 (5H, m), 6.19 (IH, d, 7= 7.3 Hz), 4.92 - 4.96 (IH, m), 3.53 - 3.73 (2H, m), 2.58 (3H, s). LCMS (M+H)+ m/z 420 (t = 1.99 min.).
Example 24
Figure imgf000093_0002
(S)-2-{4-[2-(3-Chloro-phenyl)-l-hydroxymethyl-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl}-7-methyl-3H-benzimidazole-5-carbonitrile: 1H NMR (400 MHz, CD3OD) δ 7.86 (IH, s), 7.10 - 7.33 (6H, m), 6.12 (IH, d, 7= 7.6 Hz), 4.01 - 4.05 (IH, m), 3.75 (2H, d, 7 = 4.9 Hz), 3.15 (IH, dd, 7= 4.9, 13.5 Hz), 2.86 - 3.00 (IH, m), 2.63 (3H, s). LCMS (M+H)+ m z 434 (t = 1.81 min.). Example 25
Figure imgf000094_0001
(±)-2- { 4- [2-(3 -Bromo-4-methoxy-phenyl)-2-hydroxy-εthylamino] -2-oxo- 1 ,2-dihydro- ρyridin-3-yl}-7-methyl-3H-benzimidazole-5-carbonitrile: 1H NMR (300 MHz,
CD3OD) δ 7.86 (IH, s), 7.75 (IH, s), 7.67 (IH, s), 7.28 - 7.40 (2H, m), 6.95 (IH, d, 7 = 8.5 Hz), 6.24 (IH, d, 7= 7.4 Hz), 4.93 (IH, m), 3.65 - 3.97 (2H, m), 3.82 (3H, s), 2.57 (3H, s). LCMS (M+H)+ m/z 494 (t = 2.10 min.).
Example 26
Figure imgf000094_0002
(±)-2- { 4- [2-(3 -Fluoro-phenyl)-2-hydroxy-ethylamino] -2-oxo- 1 ,2-dihydro-pyridin-3 - yl}-7-methyl-3H-benzimidazole-5-carbonitrile: 1H NMR (400 MHz, CD3OD) δ 7.70 (IH, s), 7.49 (IH, s), 7.22 - 7.32 (4H, m), 6.92 (IH, d, 7 = 8.9 Hz), 6.17 (IH, d, 7 = 7.2 Hz), 4.92 (IH, t, 7 = 6.3 Hz), 3.66 (2H, d, 7 = 5.9 Hz), 2.56 (3H, s). LCMS (M+H)+ m z 404 (t = 1.65 min.).
Example 27
Figure imgf000094_0003
(±)-2-{4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carbonitrile: 1H NMR (300 MHz, CD3OD) δ 7.92 (IH, s), 7.72 (IH, s), 7.26 - 7.50 (5H, m), 6.19 (IH, d, 7= 7.1 Hz), 4.93 (IH, t, 7 = 4.3 Hz), 3.47 - 3.73 (2H, m), 2.58 (3H, s). LCMS (M+H)+ m/z 464 (t = 2.00 min.).
Example 28
Figure imgf000095_0001
(S)-2-[4-(2-Hydroxy-2-phenyl-ethylamino)-2-oxo- 1 ,2-dihydro-pyridin-3-yl]-7- methyl-3H-benzimidazole-5-carbonitrile: 1H NMR (400 MHz, CD3OD) δ 8.04 (IH, s), 7.74 (IH, s), 7.24 - 7.51 (6H, m), 6.22 (IH, d, 7 = 7.5 Hz), 5.00 (IH, m), 3.64 - 3.74 (2H, m), 2.60 (3H, s). LCMS (M+H)+ m/z 386 (t = 1.65 min.).
Examples 29-35 were prepared from commercially or readily available diamines which were prepared and condensed with 4-iodo-2-methoxy-pyridine-3-carbaldehyde as described in Scheme HI.
Example 29
Figure imgf000095_0002
(±)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4,5,6-trifluoro-lH- benzimidazol-2-yl)-lH-pyridin-2-one: Η NMR (500 MHz, CD3COCD3) δ 13.26 (IH, broad s), 10.93 (IH, broad s), 10.27 (IH, broad s), 7.76 (IH, s), 7.54 - 7.56 (2H, m), 7.41 - 7.42 (IH, m), 7.06 (IH, d, 7 = 8.2 Hz), 6.29 (IH, d, 7 = 7.5 Hz), 5.05 - 5.07 (IH, m), 3.87 (3H, s), 3.74 - 3.79 (IH, m), 3.65 - 3.69 (IH, m).
Example 30
Figure imgf000096_0001
(±)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4,6-dibromo-lH- benzimidazol-2-yl)-lH-pyridin-2-one: Η NMR (500 MHz, CD3COCD3) δ 13.18 (IH, broad s), 11.18 (IH, broad s), 10.20 (IH, broad s), 7.92 (IH, s), 7.78 (IH, s), 7.52 - 7.55 (3H, m), 7.03 (IH, d, 7 = 8.5 Hz), 6.28 (IH, d, 7 = 7.4 Hz), 5.06 - 5.08 (IH, m), 3.86 (3H, s), 3.73 - 3.77 (IH, m), 3.66 -3.70 (IH, m).
Example 31
Figure imgf000096_0002
(±)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(5,6-dichloro-lH- benzimidazol-2-yl)-lH-pyridin-2-one: XH NMR (500 MHz, CD3COCD3) δ 7.79 (3H, m), 7.53 (IH, dd, 7 = 2.0, 8.4 Hz), 7.41 - 7.44 (IH, m), 7.08 (IH, d, 7 = 8.4 Hz), 6.29
(IH, d, 7 = 7.5 Hz), 5.05 - 5.08 (IH, m), 3.89 (3H, s), 3.74 - 3.78 (IH, m), 3.70 - 3.72 (IH, m). Example 32
Figure imgf000097_0001
(+)-3-(lH-Benzimidazol-2-yl)-4-[2-(3-bromo-phenyl)-2-hydroxy-ethylamino]-lH- pyridin-2-one: Η NMR (500 MHz, CD3COCD3) δ 7.80 (IH, s), 7.64 - 7.69 (2H, m), 7.60 (IH, d, 7= 7.7 Hz), 7.46 - 7.48 (2H, m), 7.33 (IH, t, 7= 7.8 Hz), 7.19 - 7.22 (2H, m), 6.59 (IH, d, 7= 4.5 Hz), 5.12 - 5.14 (IH, m), 5.13 (IH, dd, 7= 4.5, 7.2 Hz), 3.82 (IH, dd, 7 = 4.5, 13.6 Hz), 3.71 (IH, dd, 7= 7.2, 13.6 Hz).
Example 33
Figure imgf000097_0002
3-(lH-Benzimidazol-2-yl)-4-[(pyridin-2-ylmethyl)-amino]-lH-pyridin-2-one: 1H NMR (500 MHz, CD3COCD3) δ 8.96 - 8.98 (IH, m), 8.46 - 8.49 (IH, m), 8.03 (IH, d, 7 = 8.0 Hz), 7.93 (IH, t, 7 = 6.4 Hz), 7.69 - 7.73 (2H, m), 7.53 (IH, d, 7 = 7.5 Hz), 7.28 - 7.32 (2H, s), 6.34 (IH, d, 7 = 7.5 Hz), 5.29 (2H, m).
Example 34
Figure imgf000097_0003
(S)-3-(lH-Benzimidazol-2-yl)-4-(l-hydroxymethyl-2-phenyl-ethylamino)-lH- pyridin-2-one: 1H NMR (500 MHz, CD3COCD3) δ 7.67 - 7.69 (2H, m), 7.15 - 7.44 (8H, m), 6.31 (IH, d, 7= 7.5 Hz), 4.10 - 4.13 (IH, m), 3.76 - 3.82 (2H, m), 3.23 (IH, dd, 7 = 5.6, 13.7 Hz), 3.04 (IH, dd, 7 = 8.1, 13.7 Hz).
Example 35
Figure imgf000098_0001
(±)-3-(lH-Benzimidazol-2-yl)-4-[2-(3-bromo-4-methoxy-phenyl)-2-hydroxy- ethylamino]-lH-pyridin-2-one: 1H NMR (500 MHz, CD3COCD3) δ 7.78 (IH, s), 7.68 (2H, s), 7.50 - 7.55 (2H, m), 7.21 - 7.24 (2H, m), 7.06 (IH, d, 7= 8.4 Hz), 6.44 (IH, d, 7 = 7.5 Hz), 5.08 (IH, dd, 7 = 4.6, 7.2 Hz), 3.87 (3H, s), 3.79 (IH, dd, 7 = 4.6, 13.4 Hz), 3.71 (IH, dd, 7 = 7.2, 13.4 Hz).
Examples 36 - 43 were prepared according to Scheme V.
Example 36 (General procedure for examples 36-43)
Figure imgf000098_0002
(S)-4- { 2- [4-( 1 -Hydroxymethyl-2-phenyl-ethylamino)-2-oxo- 1 ,2-dihydro-pyridin-3- yl]-7-methyl-3H-benzimidazol-5-yl } -piperazine- 1 -carboxylic acid wøpropylamide: To a solution of (S)-4-(l-hydroxymethyl-2-phenyl-ethylamino)-3-(4-methyl-6- piperazin-l-yl-lH-benzimidazol-2-yl)-lH-pyridin-2-one (30 mg, 0.063 mmol) in methanol (2 mL) was added wøpropyl isocyanate (2 drops). The reaction mixture was stirred at room temperature for 5 min. Concentration gave the residue, which was purified by prep. HPLC to yield the title compound (23.8 mg, 69%). Η NMR (400 MHz, CD3OD) δ 7.55 (IH, s), 7.14 - 7.28 (7H, m), 6.12 (IH, d, 7 = 7.8 Hz), 4.01 - 4.03 (IH, m), 3.92 (quintet, 7 = 6.6 Hz), 3.80 (4H, m), 3.76 (IH, dd, 7 = 4.8, 11.2 Hz), 3.70 (IH, dd, 7 = 5.2, 11.2 Hz), 3.61 - 3.64 (4H, m), 3.09 (IH, dd, 7= 5.6, 13.7 Hz), 2.91 (IH, dd, 7 = 8.0, 13.7 Hz), 2.64 (3H, s), 1.19 (6H, d, 7= 6.8 Hz). LCMS (M+H)+ m/z 545 (t = 1.99 min.).
Example 37
Figure imgf000099_0001
(S)-4- { 2- [4-( 1 -Hydroxymethyl-2-phenyl-ethylamino)-2-oxo- 1 ,2-dihydro-pyridin-3-yl] -7- methyl-3H-benzimidazol-5-yl} -piperazine- 1 -carboxylic acid ethylamide: 1H NMR (400 MHz, CD3OD) δ 7.55 (IH, s), 7.12 - 7.28 (7H, m), 6.12 (IH, d, 7 = 7.8 Hz), 4.01 - 4.05 (IH, m), 3.62 - 3.81 (10H, m), 3.24 (2H, q, 7= 7.2 Hz), 3.08 (IH, dd, 7= 5.6, 13.7 Hz), 2.91 (IH, dd, 7= 8.0, 13.7 Hz), 2.65 (3H, s), 1.14 (3H, t, 7 = 7.2 Hz). LCMS (M+H)+ m/z 531 (t = 1.93 min.).
Example 38
Figure imgf000099_0002
(S)-4-( 1 -Hydroxymethyi-2-phenyl-ethylamino)-3- { 4-methyl-6- [4-( 1 -phenyl-methanoyl)- piperazin-l-yl]-lH-benzimidazol-2-yl}-lH-pyridin-2-one: To a solution of (S)-4-(l- hydroxymethyl-2-phenyl-ethylamino)-3-(4-methyl-6-piperazin- 1 -yl- 1 H-benzimidazol-2- yl)-lH-pyridin-2-one (30 mg, 0.063 mmol) in methanol (2 mL) was added benzoyl chloride (1 drop). The reaction mixture was stirred for 5 min. and concentrated. The residue was purified by prep. HPLC to yield the title compound (11 mg, 33%). 1H NMR (400 MHz, CD3OD) δ 7.48 - 7.54 (5H, m), 7.37 (IH, s), 7.13 - 7.26 (7H, m), 6.08 (IH, d, 7 = 7.7 Hz), 3.79 - 4.02 (5H, m), 3.76 (IH, dd, 7 = 4.7, 11.2 Hz), 3.67 (IH, dd, 7 = 5.7, 11.2 Hz), 3.54 (4H, m), 3.03 (IH, dd, 7 = 5.7, 13.7 Hz), 2.89 (IH, dd, 7= 8.0, 13.7 Hz), 2.62 (3H, s). LCMS (M+H)+ m/z 563 (t = 2.19 min.).
Example 39
Figure imgf000100_0001
(S)-4-(l-Hydroxymethyl-2-phenyl-ethylamino)-3-[6-(4-wopropyl-piperazin-l-yl)-4- methyl- lH-benzimidazol-2-yl]-lH-pyridin-2-one: To a solution of (S)-4-(l- hydroxymethyl-2-phenyl-ethylamino)-3-(4-methyl-6-piperazin-l-yl-lH-benzimidazol-2- yl)-lH-pyridin-2-one (25 mg, 0.054 mmol) in methanol (0.5 mL) was added acetone (0.25 mL) and 1 M THF solution of NaCNBH3 (0.2 mL). The reaction mixture was stirred at room temperature for 1 h and concentrated in vacuo. The residue was purified by prep. HPLC to yield the title compound (12 mg, 44%). 1H NMR (400 MHz, CD3OD) δ 7.10 - 7.24 (6H, m), 7.08 (IH, s), 7.06 (IH, s), 6.06 (IH, d, 7 = 7.9 Hz), 3.58 - 4.01 (10H, m), 3.34 (IH, m), 3.14 (IH, m), 2.98 (IH, dd, 7= 5.9, 13.7 Hz), 2.83 (IH, dd, 7= 7.9, 13.7 Hz), 2.60 (3H, s), 1.44 (6H, d, 7 = 6.7 Hz). LCMS (M+H)+ m/z 501 (t = 1.80 min.). Example 40
Figure imgf000101_0001
(S)-3-[6-(4-Benzyl-piperazine-l-yl)-4-methyl-lH-benzimidazol-2-yl]-4-(l- hydroxymethyl-2-phenyl-ethylamino)-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.50 - 7.57 (6H, m), 7.04 - 7.27 (7H, m), 6.07 (IH, d, 7 = 7.8 Hz), 4.22 (2H, s), 3.97 - 4.00 (IH, m), 3.34 - 3.77 (2H, m), 2.82 - 3.04 (10H, m), 2.58 (3H, s). LCMS (M+H)+ m/z 549 (t = 1.93 min.).
Example 41
Figure imgf000101_0002
(±)-3-[6-(4-Acetyl-piperazine-l-yl)-4-methyl-lH-benzimidazol-2-yl]-4-[2-(3-chloro- phenyl)-2-hydroxy-ethylamino]-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.50 (IH, s), 7.25 - 7.49 (5H, m), 7.20 (IH, s), 6.25 (IH, d, 7 = 8.0 Hz), 4.94 (IH, dd, 7 = 4.6, 7.1 Hz), 3.88 - 3.92 (4H, m), 3.50 - 3.65 (6H, m), 2.62 (3H, s), 2.20 (3H, s). LCMS (M+H)+ m/z 521 (t = 2.13 min.).
Example 42
Figure imgf000102_0001
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-piperazin-l-yl-lH- benzimidazol-2-yl) -lH-pyridin-2-one: *H NMR (400 MHz, CD3OD) δ 7.47 (IH, s), 7.25 - 7.38 (4H, m), 7.07 (2H, s), 6.25 (IH, d, 7 = 7.6 Hz), 4.90 (IH, m), 3.42 - 3.66 (10 H, m), 2.59 (3H, s). LCMS (M+H)+ m/z 479 (t = 1.90 min.).
Example 43
Figure imgf000102_0002
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[6-(4-wσpropyl-piperazine-l- yl)-4-methyl-lH-benzimidazol-2-yl]-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.47 (IH, s), 7.25 - 7.38 (4H, m), 7.06 (2H, s), 6.26 (IH, d, 7 = 7.6 Hz), 4.91 - 4.93 (IH, m), 3.89 - 3.92 (2H, m), 3.51 - 3.64 (5H, m), 3.31 - 3.37 (2H, m), 3.09 - 3.30 (2H, m), 2.59 (3H, s), 1.44 (6H, d, 7 = 6.6 Hz). LCMS (M+H)+ m/z 521 (t = 1.95 min.).
Example 44
Figure imgf000102_0003
(S)-6-( 1 -Hydroxymethyl-2-phenyl-ethylamino)-5-(6-imidazol- 1 -yl-4-methyl- 1 H- benzimidazol-2-yl)-3H-pyrimidin-4-one:
(S)-2-[6-Chloro-5-(6-imidazol- 1 -yl-4-methyl- 1 H-benzimidazol-2-yl)-pyrimidin-4- ylamino]-3-phenyl-propan-l-ol: To a solution of 2-(4,6-dichloro-pyrimidin-5-yl)-6- imidazol-l-yl-4-methyl-lH-benzimidazole (40 mg, 0.16 mmol) in wopropanol (5 mL) was added (S)-(-)-2-amino-3-phenyl-propanol (35 mg, 0.23 mmol) and triethylamine (0.5 mL). The reaction mixture was heated to 80°C for 4 h, cooled to room temperature and concentrated with high vacuum. The crude product was used for the next step without purification. LCMS (M+H)+ m/z 460 (t = 2.13 min.).
(S)-6-(l -Hydroxymethyl-2-phenyl-ethylamino)-5-(6-imidazol- 1 -yl-4-methyl- 1 H- benzimidazol-2-yl)-3H-pyrimidin-4-one: To a solution of (S)-2-[6-chloro-5-(6- imidazol-l-yl-4-methyl-lH-benzimidazol-2-yl)-pyrimidin-4-ylamino]-3-phenyl- propan-1-ol in 4 N HCl (0.5 mL) and acetic acid (0.5 mL) was added two drops of water. The reaction mixture was heated to 100°C for 8 h, cooled to room temperature, and neutralized with ammonia in methanol. After concentration, the residue was purified by prep. HPLC to yield the title compound (26 mg, 37% for two steps). *H NMR (400 MHz, CD3OD) δ 9.41 (IH, narrow t, 7 = 1.5 Hz), 8.06 (IH, narrow dd, 7 = 1.6, 1.9 Hz), 7.93 91H, s), 7.77 (IH, narrow dd, 7= 1.6, 1.8 Hz), 7.73 (IH, narrow d, 7= 1.9 Hz), 7.11 - 7.21 (6H, m), 4.70 - 4.74 (IH, m), 3.74 (2H, d, 7 = 4.7 Hz), 3.11 (IH, dd, 7 = 6.1, 13.5 Hz), 3.00 (IH, dd, 7 = 7.8, 13.5 Hz), 2.70 (3H, s). LCMS (M+H)+ m/z 442 (t = 2.17 min.).
Examples 45 - 383 were prepared according to the general methods described above (Scheme HT)
LCMS conditions: a) YMC C18 S5 4.6 x 50 mm; 0 - 100% gradient over 4 min*; 4 rnL/min flow rate b) YMC ODS-A C18 S7 3.0 x 50 mm; 0 - 100% gradient over 2 min*; 5 mL min flow rate c) YMC C18 S5 4.5 x 50 mm; 0 - 100% gradient over 8 min*; 2.5 mL/min flow rate d) YMC C18 S7 3.0 x 50 mm; 0 - 100% gradient over 3 min*; 5 mL/min flow rate e) YMC ODSA S3 6.0 xl50 mm; 0 - 100% gradient over 5 min*; 1.5 mL min flow rate f) PHS-PRJJVIESPHERE C184.6 x 30 mm; 0 - 100% gradient over 2 min*; 5 mL min flow rate g) YMC C18 S7 3.0 x 50 mm; 0 - 100% gradient over 4 min*; 5 mlJmin flow rate h) YMC ODS-A C18 S7 3.0 x 50 mm; 0 - 100% gradient over 2 min*; 5 mL min flow rate i) YMC ODS-A C18 S7 3.0 x 50 mm; 0 - 100% gradient over 1.5 min*; 5 mL/min flow rate j) YMC Xterra C18 S7 3.0 x 50 mm; 0 - 100% gradient over 2 min*; 5 mL/min flow rate k) YMC Pro-ODS C18 S54.6x33 mm; 0 - 100% gradient over 3 min*; 4 mL/min flow rate 1) YMC ODS-A C18 S7 3.0 x 50 mm; 0 - 100% gradient over 4 min*; 4 mL/min flow rate * Gradient begin with 10% methanol 90% water (0.1% TFA) and end with 90% methanol/10% water (0.1% TFA)
Figure imgf000104_0001
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Figure imgf000155_0001
Figure imgf000156_0001
Intermediate Imidate Formation (Scheme IN, 13)
Figure imgf000157_0001
(S)-2-[4-(l-Hydroxymethyl-2-phenyl-ethylamino)-2-oxo-l,2-dihydro-pyridin-3-yl]-7- methyl-3H-benzimidazole-5-carboximidic ethyl ester: To a suspension of (S)-2-[4-(l- hydroxymethyl-2-phenyl-ethylamino)-2-oxo-l,2-dihydro-pyridin-3-yl]-7-methyl-3H- benzimidazole-5-carbonitrile (0.8 g, 2.0 mmol) in ethanol (anhydrous, 80 mL) was bubbled HCl (anhydrous) at 0°C until saturation. The mixture became a clear solution after a few minutes of bubbling and the solution was stirred at room temperature for 14 h. After concentration in vacuo, the crude product (0.89 g, 100%) was directly used for the next step without purification. LCMS (M+H)+ m/z 446 (t = 1.55 min).
Figure imgf000157_0002
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboximidic ethyl ester: The title compound was prepared using the General Procedure for Imidate Formation. LCMS (M+H)+ m/z 466 (t = 1.43 min.).
General Procedure for Examples 384 - 397 - Imidazoline Formation (Scheme IN, 14) Example 384
Figure imgf000158_0001
(S)-3-[6-(4,5-Dihydro-lH-imidazol-2-yl)-4-methyl-lH-benzimidazol-2-yl]-4-(l- hydroxymelhyl-2-phenyl-ethylamino)-lH-pyridin-2-one: The crude imidate ester (60 mg, 0.135 mmol) was diluted with methanol followed by addition of ethylenediamine (24 mg, 0.40 mmol). The reaction mixture was heated to reflux for 6 h. After concentration in vacuo, the residue was purified by prep. HPLC to yield the title compound (37 mg, 62%). Η NMR (300 MHz, CD3OD) δ 7.94 (IH, s), 7.50 (IH, s), 7.12 - 7.30 (6H, m), 6.16 (IH, d, J= 1.1 Hz), 4.04 - 4.10 (5H, m), 3.75 - 3.77 (2H, m), 3.15 (IH, dd, J= 5.2, 13. 6 Hz), 2.96 (IH, dd, /= 8.1, 13.6 Hz), 2.67 (3H, s). LCMS (M+H)+ m/z 443 (t = 1.50 min.).
Example 385
Figure imgf000158_0002
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[6-(4,5-dihydro-lH-imidazol-2- yl)-4-methyl-lH-benzimidazol-2yl]-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.93 (IH, s), 7.54 (IH, s), 7.23 - 7.45 (5H, m), 6.26 (IH, d, J = 7.6 Hz), 5.01 (IH, t, J = 6.3 Hz), 4.01 (4H, s), 3.78 (IH, dd, J = 4.7, 13.5 Hz), 3.67 (IH, dd, J = 6.6, 13.5 Hz), 2.66 (3H, s). LCMS (M+H)+ m/z 463 (t = 1.54 min Example 386
Figure imgf000159_0001
4-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-3-[4-methyl-6-(4S-methyl-4,5- dihydro-lH-imidazol-2-yl)-lH-benzoimidazol-2-y]]-lH-pyridin-2-one and 4-[2-(3- Chloro-phenyl)-2S-hydroxy-ethylamino]-3-[4-methyl-6-(4S-methyl-4,5-dihydro-lH- imidazol-2-yl)-lH-benzoimidazol-2-yl]-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.90 (2H, s), 7.53 (2H, s), 7.22 - 7.47 (10H, m), 6.23 (2H, d, J= 7.5 Hz), 5.00 (2H, t, J= 6.4 Hz), 4.49 - 4.57 (2H, m), 4.21 (2H, t, J = 11.0 Hz), 3.61 - 3.78 (6H, m), 2.63 (6H, s), 1.48 (6H, d, J= 6.3 Hz). LCMS (M+H)+ m/z 477 (t = 1.71 min.).
Example 387
Figure imgf000159_0002
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(l-methyl-4,5- dihydro-lH-imidazol-2-yl)-lH-benzimidazol-2yl]-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.94 (IH, s), 7.57 (IH, s), 7.23 - 7.52 (5H, m), 6.22 (IH, d, J = 7.5 Hz), 4.99 (IH, m), 4.00 - 4.15 (2H, m), 3.57 - 3.75 (4H, m), 2.77 (3H, s), 2.61 (3H, s). LCMS (M+H)+ m/z 477 (t = 1.60 min.).
Example 388
Figure imgf000160_0001
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[6-(4,4-dimethyl-4,5-dihydro- lH-imidazol-2-yl)-4-methyl-lH-benzimidazol-2yl]-lH-pyridin-2-one: 'H NMR (300 MHz, CD3OD) δ 7.89 (IH, s), 7.52 (IH, s), 7.22 - 7.45 (5H, m), 6.19 (IH, d, J = 7.4 Hz), 4.99 (IH, t, J = 6.2 Hz), 3.84 (2H, s), 3.78 (IH, dd, J = 4.4, 13.4 Hz), 3.60 (IH, dd, /= 6.7, 13.4 Hz), 2.61 (3H, s), 1.55 (6H, s). LCMS (M+H)+ m z 491 (t = 1.73 min.).
Example 389
Figure imgf000160_0002
2-(2-{4-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazol-5-yl)-4,5-dihydro-lH-imidazole-4S-carboxylic acid and 2-(2-{4-[2-(3-Chloro-phenyl)-2S-hydroxy-ethylamino]-2-oxo-l ,2-dihydro- pyridin-3-yl}-7-methyl-3H-benzimidazol-5-yl)-4,5-dihydro-lH-imidazole-4S- carboxylic acid: 1H NMR (300 MHz, CD3OD) δ 7.88 (2H, s), 7.52 (2H, s), 7.22 - 7.44 (10H, m), 6.17 (2H, d, /= 7.4 Hz), 5.05 (2H, m), 4.23 - 4.38 (4H, m), 3.56 - 3.72 (6H, m), 2.58 (6H, s). LCMS (M+H)+ m/z 507 (t = 1.64 min.). Example 390
Figure imgf000161_0001
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(l,4,5,6-tetrahydro- pyrimidin-2-yl)-lH-benzimidazol-2-yl]-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.90 (IH, s), 7.53 (IH, s), 7.22 - 7.47 (5H, m), 6.23 (IH, d, 7= 7.6 Hz), 5.00 (IH, t, J = 6.3 Hz), 4.49 - 4.57 (IH, m), 3.60 - 3.77 (5H, m), 3.24 - 3.34 (2H, m), 2.63 (3H, s). LCMS (M+H)+ m/z 477 (t = 1.59 min.).
Example 391
Figure imgf000161_0002
(±)-4-[2-(3-Chloro-4-methoxy-phenyl)-2-methoxy-ethylamino]-3-[6-(4,5-dihydro- lH-imidazol-2-yl)-4-methyl-lH-benzimidazol-2-yl]-lH-ρyridin-2-one: *H NMR (300 MHz, CD3OD) δ 7.88 (IH, s), 7.46 (IH, s), 7.44 (IH, s), 7.31 (IH, d, /= 6.4 Hz), 7,28 (IH, d, J = 5.6 Hz), 7.01 (IH, d, J = 6.4 Hz), 6.19 (IH, d, / = 5.6 Hz), 4.49 (IH, dd, J = 3.4, 5.0 Hz), 4.09 (4H, s), 3.82 (3H, s), 3.36 (3H, s), 3.63 - 3.67 (2H, m), 2.62 (3H, s). LCMS (M+H)+ m/z 507 (t = 1.72 min.). Example 392
Figure imgf000162_0001
(±)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hj*droxy-ethj amino]-3-[6-(4,5-dihydro-lH- imidazol-2-yl)-4-methyl- 1 H-benzimidazol-2-yl] - 1 H-pyridin-2-one: The title compound was prepared following hydrolysis of the benzylic chloride to the hydroxyl. 1H NMR (400 MHz, CD3OD) δ 7.90 (IH, s), 7.67 (IH, s), 7.48 (IH, s), 7.38 (IH, dd, 7 = 2.0, 8.5 Hz), 7.31 (IH, d, 7= 7. 6 Hz), 6.95 (IH, d, 7= 8.5 Hz), 6.26 (IH, d, 7= 7.6 Hz), 4.94 (IH, m), 4.10 (4H, s), 3.80 (3H, s), 3.62 - 3.74 (2H, m), 2.62 (3H, s). LCMS (M+H)+ m/z 537 (t = 1.49 min.).
Example 393
Figure imgf000162_0002
(S)-3-[6-(4,4-Dimethyl-4,5-dihydro-lH-imidazol-2-yl)-4-methyl-lH-benzimidazol-2- yl]-4-(l-hydroxymethyl-2-phenyl-ethylamino)-lH-pyridin-2-one: ]H NMR (300 MHz, CD3OD) δ 7.94 (IH, s), 7.50 (IH, s), 7.12 - 7.30 (6H, m), 6.15 (IH, dd, 7= 7.7 Hz), 4.02 - 4.10 (IH, m), 3.85 (2H, s), 3.75 - 3.77 (2H, m), 2.96 - 3.17 (2H, m), 2.67 (3H, s), 1.56 (6H, s). LCMS (M+H)+ m/z 471 (t = 1.62 min.). Example 394
Figure imgf000163_0001
(S)-2- { 2- [4-( 1 S-Hydroxymethyl-2-phenyl-ethylamino)-2-oxo- 1 ,2-dihydro-pyridin-3- yl]-7-methyl-3H-benzimidazol-5-yl}-4,5-dihydro-lH-imidazole-4S-carboxylic acid. 1H NMR (300 MHz, CD3OD) δ 8.03 (IH, s), 7.60 (IH, s), 7.11 - 7.29 (6H, m), 6.15 (IH, d, 7 = 7.3 Hz), 5.07 (IH, dd, 7 = 6.9, 11.8 Hz), 4.25 - 4.41 (2H, m), 4.05 - 4.07 (IH, m), 3.70 - 3.82 (2H, m), 3.12 (IH, dd, 7 = 5.5, 13.6 Hz), 2.95 (IH, dd, 7 = 8.0, 13.6 Hz), 2.69 (3H, s). LCMS (M+H)+ m/z 487 (t = 1.35 min.).
Example 395
Figure imgf000163_0002
(S)-4-(l-Hydroxymethyl-2-phenyl-ethylamino)-3-[4-methyl-6(l,4,5,6-tetrahydro- pyrimidin-2-yl)-lH-benzimidazol-2-yl]-lH-pyridin-2-one: ]H NMR (300 MHz, CD3OD) δ 7.76 (IH, s), 7.32 (IH, s), 7.09 - 7.29 (6H, m), 6.15 (IH, d, 7= 7.6 Hz), 4.02 - 4.07 (IH, m), 3.71 - 3.79 (2H, m),*3.61 (4H, t, 7= 5.6 Hz), 3.14 (IH, dd, 7 = 5.3, 13.6 Hz), 2.95 (IH, dd, 7= 8.1, 13.6 Hz), 2.65 (3H, s), 2.09 - 2.16 (2H, m). LCMS (M+H)+ m/z 457 (t = 1.49 min.). Example 396
Figure imgf000164_0001
(S)-4-(l-Hydroxymethyl-2-phenyl-ethylamino)-3-[4-methyl-6-(l-methyl-4,5-dihydro- lH-imidazol-2-yl)-lH-benzoimidazo-2-yl]-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.77 (IH, s), 7.10 - 7.30 (7H, m), 6.16 (IH, d, 7 = 7.6 Hz), 3.97 - 4.20 (5H, m), 3.73 - 3.77 (2H, m), 3.26 (3H, s), 3.15 (IH, dd, 7 = 5.3, 13.6 Hz), 2.96 (IH, dd, 7 = 8.0, 13.6 Hz), 2.68 (3H, s). LCMS (M+H)+ m/z 457 (t = 1.53 min.)
Example 397
Figure imgf000164_0002
(S)-3-[6-(4,5-Dihydro-lH-imidazol-2-yl)-4-methyl-lH-benzimidazol-2-yl]-4-(2- hydroxy-2-phenyl-ethylamino)-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.91 (IH, s), 7.50 (IH, s), 7.48 (2H, s), 7.24 - 7.35 (4H, m), 6.25 (IH, d, 7= 7.6 Hz), 5.01 (IH, dd, 7 = 4.6, 7.0 Hz), 4.10 (4H, s), 3.65 - 3.76 (2H, m), 2.64 (3H, s). LCMS (M+H)+ m/z 429 (t = 1.48 min.).
Example 398
Figure imgf000165_0001
(S)-4-(l-Hydroxymethyl-2-phenyl-ethylamino)-3-[6-(lH-imidazol-2-yl)-4-methyl- lH-benzimidazol-2-yl]-lH-ρyridin-2-one: To a solution of the imidate ester (150 mg, 0.31 mmol) in methanol (10 mL) was added aminoacetaldehyde diethyl acetal (97 mg, 0.93 mmol). The mixture was stirred at room temperature for 14 h. The solvent was removed in vacuo and the residue was treated with 60% HClO4 (5 mL) at room temperature for 14 h. The reaction mixture was then neutralized with ammonium hydroxide (cone). The inorganic salt was filtered. The filtrate was concentrated and purified by prep. HPLC to yield the title compound (37 mg, 27%) as a white solid. 1H NMR (300 MHz, CD3OD) δ 7.94 (IH, narrow d, 7= 1.3 Hz), 7.58 (2H, s), 7.53 (IH, narrow d, 7 = 0.6 Hz), 7.12 - 7.31 (6H, m), 6.16 (IH, d, 7 = 7.7 Hz), 4.04 - 4.08 (IH, m), 3.76 - 3.79 (2H, m), 3.16 (IH, dd, 7 = 5.4, 13.6 Hz), 3.97 (IH, dd, 7= 8.1, 13.6 Hz), 2.69 (3H, s). LCMS (M+H)+ m/z 441 (t = 1.60 min.).
General Procedure for Amidine Formation, Examples 399 - 412 (Scheme IN, 16) Example 399
Figure imgf000165_0002
(S)-2-[4-(l-Hydroxymethyl-2-phenyl-ethylamino)-2-oxo-l,2-dihydro-pyridin-3-yl]- 7,N-dimethyl-3H-benzimidazole-5-carboxamidine: The crude imidate ester (60 mg, 0.135 mmol) was diluted with methanol (10 mL), then methylamine (2.0 M methanol solution, 0.5 mL, excess) was added to the solution. After stirring for 2 h, the reaction mixture was concentrated and purified by prep. HPLC to yield the title compound (32 mg, 55%) as a white solid. 1H NMR (300 MHz, CD3OD) δ 7.82 (IH, s), 7.38 (IH, s), 7.10 - 7.30 (6H, m), 6.15 (IH, 7 = 7.7 Hz), 4.03 - 4.07 (IH, m), 4.72 - 4.76 (2H, m), 3.06 - 3.18 (IH, m), 3.12 (3H, s), 2.96 (IH, dd, 7 = 8.0, 13.5 Hz), 2.67 (3H, s). LCMS (M+H)+ m/z 431 (t = 1.34 min.).
Example 400
Figure imgf000166_0001
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl }-7N-dimethyl-3H-benzimidazole-5-carboxamidine: 1H NMR (300 MHz, CD3OD) δ 7.81 (IH, s), 7.54 (IH, s), 7.23 - 7.54 (5H, m), 6.26 (IH, d, 7 = 7.6 Hz), 5.01 (IH, t, 7 = 4.8 Hz), 3.77 (IH, dd, 7 = 4.5, 13.4 Hz), 3.67 (IH, dd, 7= 6.6, 13.4 Hz), 3.12 (3H, s), 2.66 (3H, s). LCMS (M+H)+ m/z 451 (t = 1.32 min.).
Example 401
Figure imgf000166_0002
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxamidine: 1H NJVIR (300 MHz, CD3OD) δ 7.90 (IH, s), 7.26 -7.57 (6H, m), 6.25 (IH, d, 7 = 7.6 Hz), 5.01 (IH, t, 7 = 6.4 Hz), 3.75 (IH, dd, 7 = 4.8, 13.4 Hz), 3.66 (IH, dd, 7 = 6.7, 13.4 Hz), 2.62 (3H, s). LCMS (M+H)+ m/z 437 (t = 1.59 min.). Example 402
Figure imgf000167_0001
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl } -7,NN,-trimethyl-3H-benzimidazole-5-carboxamidine: Η ΝMR (300 MHz, CD3OD) δ 7.63 (IH, s), 7.54 (IH, s), 7.18 - 7.41 (5H, m), 6.26 (IH, d, 7= 7. 6 Hz), 5.01 (IH, t, 7 = 6.3 Hz), 3.77 (IH, dd, 7 = 4.7, 13.5 Hz), 3.67 (IH, dd, 7 = 6.6, 13.5 Hz), 3.34 (6H, s), 2.66 (3H, s). LCMS (M+H)+ m/z 465 (t = 1.57 min.).
Example 403
Figure imgf000167_0002
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-N-cyclopropyl-7-methyl-3H-benzimidazole-5-carboxamidine: 1H ΝMR (300 MHz, CD3OD) δ 7.78 (IH, s), 7.54 (IH, s), 7.22 - 7.41 (5H, m), 6.25 (IH, d, 7 = 7.6 Hz), 5.01 (IH, t, 7 = 6.4 Hz), 3.76 (IH, dd, 7 = 4.7, 13.5 Hz), 3.67 (IH, dd, 7 = 6.6, 13.5 Hz), 2.77 - 2.83 (IH, m), 2.65 (3H, s), 1.03 - 1.10 (2H, m), 0.86 - 0.92 (2H, m). LCMS (M+H)+ m/z 477 (t = 1.43 min.). Example 404
Figure imgf000168_0001
(±)-2-{4-[2-(3-Chloro-4-methoxy-phen3d)-2-methoxy-ethylamino]-2-oxo-l,2- dihydro-pyridin-3-yl } -7,N-dimethyl-3H-benzimidazole-5-carboxamidine: 1H ΝMR (300 MHz, CD3OD) δ 7.79 (IH, narrow d, 7 = 1.0 Hz), 7.45 (IH, narrow d, 7= 1.5 Hz), 7.38 (IH, s), 7.28 - 7.31 (2H, m), 7.02 (IH, d, 7 = 6.4 Hz), 6.22 (IH, d, 7= 5.7 Hz), 4.52 (IH, t, 7= 5.9 Hz), 3.83 (3H, s), 3.60 - 3.71 (2H, m), 3.36 (3H, s), 3.13 (3H, s), 2.65 (3H, s). LCMS (M+H)+ m/z 495 (t = 1.64 min.).
Example 405
Figure imgf000168_0002
(±)-2-{4-[2-(3-Chloro-4-methoxy-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl } -7,N-dimethyl-3H-benzimidazole-5-carboxamidine: To a solution of 2- {4-[2-(3-chloro-4-methoxy-phenyl)-2-methoxy-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl}-7N-dimethyl-3H-benzimidazole-5-carboxamidine (40 mg, 0.08 mmol) in methanol (5 mL) was added 2 Ν ΝaOH (0.5 mL) and the reaction mixture was stirred at room temperature for 14 h. After concentration in vacuo, the residue was purified by prep. HPLC to yield the title compound (7.2 mg, 19%) as oil. Η ΝMR (300 MHz, CD3OD) δ 7.78 (IH, narrow d, 7 = 1.6 Hz), 7.52 (IH, narrow d, 7 = 2.1 Hz), 7.30 - 7.36 (3H, m), 6.99 (IH, d, 7 = 8.5 Hz), 6.27 (IH, d, 7 = 7.6 Hz), 4.94 (IH, t, 7 = 5.9 Hz), 3.82 (3H, s), 3.68 - 3.72 (2H, m), 3.13 (3H, s), 2.63 (3H, s). LCMS (M+H)+ m/z 481 (t = 1.44 min.). Example 406
Figure imgf000169_0001
(±)-2- { 4- [2-(3 -Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro- pyridin-3-yl}-7N-dimethyl-3H-benzimidazole-5-carboxamidine: 1H NMR (400 MHz, CD3OD) δ 7.77 (IH, s), 7.67 (IH, s), 7.29 - 7.39 (3H, m), 6.95 (IH, d, 7= 8.4 Hz), 6.23 (IH, d, 7 = 4.4 Hz), 4.92 (IH, m), 3.80 (3H, s), 3.69 (2H, m), 3.13 (3H, s), 2.61 (3H, s). LCMS (M+H)+ m/z 525 (t = 1.44 min.).
Example 407
Figure imgf000169_0002
(S)-N-Cyclopropyl-2-[4-(l -hydroxymethyl-2-phenyl-ethylamino)-2-oxo- 1 ,2-dihydro- pyridin-3-yl]-7-methyl-3H-benzimidazole-5-carboxamidine: 1H ΝMR (300 MHz, CD3OD) δ 7.81 (IH, s), 7.37 (IH, s), 7.10 - 7.30 (6H, m), 6.16 (IH, d, 7= 7.6 Hz), 4.03 - 4.08 (IH, m), 3.74 - 3.76 (2H, m), 3.15 (IH, dd, 7= 5.3, 13.6 Hz), 2.96 (IH, dd, 7= 8.1, 13.6 Hz), 2.78 - 2.84 (IH, m), 2.67 (3H, s), 1.03 - 1.10 (2H, m), 0.87 - 0.92 (2H, m). LCMS (M+H)+ m/z 457 (t = 1.51 min.). Example 408
Figure imgf000170_0001
(S)-N-Ethyl-2- [4-( 1 -hydroxymethyl-2-phenyl-ethylamino)-2-oxo- 1 ,2-dihydro-pyridin- 3-yl]-7-methyl-3H-benzimidazole-5-carboxamidine: 1H ΝMR (400 MHz, CD3OD) δ 7.82 (IH, s), 7.38 (IH, s), 7.12 - 7.30 (6H, m), 6.15 (IH, d, 7 = 7.8 Hz), 4.04 - 4.07 (IH, m), 3.75 - 3.77 (2H, m), 3.51 (2H, q, 7 = 7.3 Hz), 3.15 (IH, dd, 7= 5.4, 13.7 Hz), 2.96 (IH, dd, 7 = 8.1, 13.7 Hz), 2.67 (3H,s), 1.40 (3H, t, 7 = 7.3 Hz). LCMS (M+H)+ m/z 445 (t = 1.44 min.).
Example 409
Figure imgf000170_0002
(S)-2-{4-[2-(3-Chloro-phenyl)-l-hydroxymethyl-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl}-7,N-dimethyl-3H-benzimidazole-5-carboxamidine: Η ΝMR (400 MHz, CD3OD) δ 7.83 (IH, s), 7.38 (IH, s), 7.31 (IH, s), 7.10 - 7.24 (4H, m), 6.16 (IH, d, 7 = 7.6 Hz), 4.06 -4.09 (IH, m), 3.72 - 3.77 (2H, m), 3.16 (3H, s), 3.15 (IH, m), 2.96 (IH, dd, 7 = 8.2, 13.7 Hz), 2.67 (3H, s). LCMS (M+H)+ m/z 465 (t = 1.49 min.).
Example 410
Figure imgf000170_0003
(S)-2-{4-[2-(2-Chloro-phenyl)-l-hydroxymethyl-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl}-7,N-dimethyl-3H-benzimidazole-5-carboxamidine: 1H ΝMR (400 MHz, CD3OD) δ 7.84 (IH, s), 7.41 (IH, s), 7.32 - 7.35 (2H, m), 7.06 - 7.18 (3H, m), 6.10 (IH, d, 7 = 7.5 Hz), 4.20 - 4.24 (IH, m), 3.76 - 3.85 (2H, m), 3.30 - 3.34 (IH, m), 3.06 - 3.14 (IH, m), 3.13 (3H, s), 2.64 (3H, s). LCMS (M+H)+ m/z 465 (t = 1.48 min.).
Example 411
Figure imgf000171_0001
(±)-2- { 4- [2-(3-Bromo-phenyl)-2-hydroxy-ethylamino] -2-oxo- 1 ,2-dihydro-pyridin-3- yl}-7N-dimethyl-3H-benzimidazole-5-carboxamidine: 'H ΝMR (300 MHz, CD3OD) δ 7.80 (IH, s), 7.69 (IH, s), 7.20 - 7.45 (5H, m), 6.25 (IH, d, 7 = 7.3 Hz), 5.00 (IH, t, 7 = 5.9 Hz), 3.64 - 3.80 (2H, m), 3.12 (3H, s), 2.66 (3H, s). LCMS (M+H)+ m/z 495 (t = 1.49 min.).
Example 412
Figure imgf000171_0002
(S)-2-[4-(2-Hydroxy-2-phenyl-ethylamino)-2-oxo-l,2-dihydro-pyridin-3-yl]-7N- dimethyl-3H-benzimidazole-5-carboxamidine: 1H NMR (400 MHz, CD3OD) δ 7.78 (IH, s), 7.24 - 7.49 (7H, m), 6.21 (IH, d, 7 = 7.0 Hz), 4.00 - 5.05 (IH, m), 3.61 - 3.74 (2H, m), 3.12 (3H, s), 2.63 (3H, s). LCMS (M+H)+ m/z 417 (t = 1.28 min.).
Figure imgf000172_0001
Figure imgf000173_0001
Figure imgf000174_0001
Figure imgf000175_0001
Figure imgf000176_0001
Figure imgf000177_0001
Figure imgf000178_0001
Figure imgf000179_0001
Figure imgf000180_0001
Figure imgf000181_0002
Amide Formation Intermediate Synthesis
Figure imgf000181_0001
(±)-2-{4-[2-(3-Chloro-ρhenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxylic acid ethyl ester: A solution of (±)-2-{4- [2-(3-chloro-phenyl)-2-hydroxy-ethylamino-]-2-oxo-l,2-dihydro-pyridin-3-yl}-7- methyl-3H-benzimidazole-5-carboximidic acid ethyl ester (400 mg, 0.90 mmol) was diluted with 2 N HCl (20 mL) solution and the mixture was allowed to stir at room temperature for 14 h. After concentration to dryness, the crude product (419 mg, 100%) was used for the next step without purification. Small amount was purified by prep. HPLC to yield the title compound. 1H NMR (300 MHz, CD3OD) δ 8.11 (IH, s), 7.74 (IH, s), 7.25 - 7.54 (5H, m), 6.25 (IH, d, 7= 7.6 Hz), 4.99 (IH, t, 7= 7.2 Hz), 4.38 (2H, q, 7 = 7.1 Hz), 3.61 - 3.76 (2H, m), 2.61 (3H, s), 1.42 (3H, q, 7= 7.1 Hz). LCMS (M+H)+ m/z 467 (t = 2.23 min.).
Figure imgf000182_0001
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxylic acid: The ethyl ester (419 mg, 0.90 mmol) obtained above was diluted with methanol (15 mL) and water (5 mL) followed by addition of sodium hydroxide (180 mg, 4.5 mmol). The mixture was stirred at room temperature for 14 h. After removal of methanol, the residue was neutralized with 2 N HCl solution. The resulting slurry was filtered and washed with ice-cold water. The solid was collected after drying over high vacuum. The crude product (395 mg, 100% yield, 80% pure) was used for the next step without further purification. 'NMR (300 MHz, CD3OD) δ 8.20 (IH, s), 7.92 (IH, s), 7.27 - 7.47 (5H, m), 6.27 (IH, d, 7 = 7.6 Hz), 4.85 (IH, m), 3.63 (2H, m), 2.67 (3H, s). LCMS (M+H)+ m z 439 (t = 1.88 min.).
Example 475
Figure imgf000182_0002
(S)-2-[4-(l-Hydroxymethyl-2-phenyl-ethylamino)-2-oxo-l,2-dihydro-pyridin-3-yl]-7- methyl-3H-benzimidazole-5-carboxylic acid ethyl ester: The imidate ester (30 mg, 0.067 mmol) was dilute with methanol (10 mL) and two drops of water was added. The reaction mixture was stirred at room temperature for 20 h. After concentration in vacuo, the residue was purified by prep. HPLC to yield the title compound (18.2 mg, 61%) as a white solid. 1H NMR (300 MHz, CD3OD) δ 7.74 (IH, s), 7.11 - 7.30 (7H, m), 6.10 (IH, d, 7 = 6.9 Hz), 4.37 (2H, q, 7 = 6.6 Hz), 4.02 (IH, broad s), 3.69 - 3.81 (2H, m), 3.10 (IH, dd, 7 = 4.8, 13.5 Hz), 2.93 (IH, dd, 7= 7.8, 13.5 Hz), 2.62 (3H, s), 1.41 (3H, 17= 6.6 Hz). LCMS (M+H)+ m/z 447 (t = 1.95 min.).
Preparation of Amides Starting from (±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy- ethylamino] -2-oxo- 1 ,2-dihydro-pyridin-3-yl } -7-methyl-3H-benzimidazole-5- carboxylic acid, Examples 476 - 504, Scheme IN, 15
Example 476
Figure imgf000183_0001
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxylic acid methylamide: To a solution of the acid (50 mg, 0.11 mmol) in DMF (5 mL) was added diphenylphosphoryl azide (38 mg, 0.14 mmol). The mixture was allowed to stir for 5 min. Then methylamine (2.0 M solution in THF)(0.11 mL, 0.22 mmol) was added. The mixture was stirred at room temperature for 14 h. After removal of DMF with high vacuum, the residue was purified by prep. HPLC to yield the title compound (18 mg, 36%) as a white solid. 1H ΝMR (300 MHz, CD3OD) δ 7.89 (IH, s), 7.18 - 7.53 (6H, m), 6.23 (IH, d, 7= 7.6 Hz), 4.98 (IH, m), 3.70 (IH, dd, 7 = 4.7, 13.5 Hz), 3.61 (IH, dd, 7 = 7.0, 13.5 Hz), 2.94 (3H, s), 2.61 (3H, s). LCMS (M+H)+ m/z 452 (t = 1.57 min.).
Example 477
Figure imgf000183_0002
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxylic acid dimethylamide: 1H NMR (300 MHz, CD3OD) δ 7.52 (IH, s), 7.24 - 7.40 (5H, m), 7.15 (IH, s), 6.25 (IH, d, 7= 7.6 Hz), 4.98 (IH, dd, 7 = 5.0, 6.9 Hz), 3.70 (IH, dd, 7 = 4.8, 13.5 Hz), 3.62 (IH, dd, 7 = 7.0, 13.5 Hz), 3.3 (3H, s), 3.08 (3H, s), 2.62 (3H, s). LCMS (M+H)+ m/z 466 (t = 1.84 min.).
Example 478
Figure imgf000184_0001
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxylic acid tert-butylamide: 1H NMR (300 MHz, CD3OD) δ 7.82 (IH, s), 7.53 (IH, s), 7.48 (IH, s), 7.24 - 7.40 (4H, m), 6.25 (IH, d, 7 = 7.6 Hz), 5.00 (IH, m), 3.64 - 3.70 (2H, m), 2.62 (3H, s), 1.49 (9H, s). LCMS (M+H)+ m/z 494 (t = 1.79 min.).
Example 479
Figure imgf000184_0002
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxylic acid butyl-methyl-amide: 1H NMR (300 MHz, CD3OD) δ 7.51 (IH, s), 7.48 (IH, s), 7.24 - 7.39 (4H, m), 7.13 (IH, s), 6.22 (IH, d, 7 = 7.6 Hz), 4.95 (IH, m), 3.55 - 3.70 (3H, m), 3.34 (IH, m), 3.05 (3H, s), 2.62 (3H, s), 0.79 - 1.69 (7H, m). LCMS (M+H)+ m/z 508 (t = 1.88 min.).
Example 480
Figure imgf000185_0001
(±)-2- { 4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro-pyridin-3- yl } -7-methyl-3H-benzimidazole-5-carboxylic acid (2-moφholine-4-yl-ethyl)-amide: 1H NMR (300 MHz, CD3OD) δ 7.58 (IH, s), 7.52 (IH, s), 7.05 - 7.42 (5H, m), 6.22 (IH, d, 7 = 7.6 Hz), 4.97 (IH, dd, 7 = 4.9, 6.7 Hz), 2.60 - 4.07 (14H, m), 2.61 (3H, s). LCMS (M+H)+ m/z 551 (t = 1.49 min.).
Example 481
Figure imgf000185_0002
(±)-4-[ 1 -(2- { 4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro- pyridin-3-yl}-7-methyl-3H-benzimidazol-5-yl)-methanoyl]-piperazine-l-carboxylic acid tert-butyl ester: !H NMR (300 MHz, CD3OD) δ 7.52 (IH, s), 7.50 (IH, s), 7.24 - 7.38 (4H, m), 7.18 (IH, m), 6.22 (IH, d, 7 = 7.6 Hz), 4.97 (IH, m), 3.49 - 3.69 (10H, m), 2.62 (3H, s), 1.47 (9H, s). LCMS (M+H)+ m/z 607 (t = 1.90 min.). Example 482
Figure imgf000186_0001
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(l-piperazin-l-yl- methanoyl)-lH-benzimidazol-2-yl]-lH-pyridin-2-one: To a solution of (±)-4-[l-(2- { 4- [2-(3 -chloro-phenyl)-2-hydroxy-ethylamino] -2-oxo- 1 ,2-dihydro-pyridin-3 -yl } -7 - methyl-3H-benzimidazol-5-yl)-methanoyl] -piperazine- 1 -carboxylic acid tert-butyl ester (40 mg, 0.06 mmol) in methanol (5 mL) was added 4.0 M HCl dioxane solution (0.1 mL, excess). The reaction mixture was stirred at room temperature for 14 h. After concentration, the residue was purified by prep. HPLC to give the title compound (16 mg, 63%) as a white foam. 1H NMR (300 MHz, CD3OD) δ 7.56 (IH, s), 7.53 (IH, s), 7.23 - 7.41 (4H, m), 7.17 (IH, m), 6.26 (IH, d, 7= 7.6 Hz), 4.99 (IH, dd, 7 = 4.7, 6.4 Hz), 3.91 (4H, broad s), 3.74 (IH, dd, 7 = 4.7, 13.5 Hz), 3.65 (IH, dd, 7= 6.4, 13.5 Hz), 3.31 (4H, broad s), 2.63 (3H, s). LCMS (M+H)+ m z 507 (t = 1.38 min.).
Example 483
Figure imgf000186_0002
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxylic acid cyclopropylamide: "H NMR (300 MHz, CD3OD) δ 7.80 (IH, s), 7.21 - 7.53 (6H, m), 6.22 (IH, d, 7 = 7.6 Hz), 4.99 (IH, t, 7 = 6.4 Hz), 3.59 - 3.76 (2H, m), 2.83 - 2.90 (IH, m), 2.60 (3H, s), 0.81 - 0.89 (2H, m), 0.64 - 0.73 (2H, m). LCMS (M+H)+ m/z 478 (t = 1.60 min.). Example 484
Figure imgf000187_0001
(±)-2- { 4- [2-(3 -Chloro-phenyl)-2-hydroxy-ethyl amino] -2-oxo- 1 ,2-dihydro-pyridin-3 - yl}-7-methyl-3H-benzimidazole-5-carboxylic acid cyclopentylamide: 'H NMR (300 MHz, CD3OD) δ 7.97 (IH, s), 7.16 - 7.60 (6H, m), 6.22 (IH, d, 7 = 7.6 Hz), 4.35 (IH, m), 3.58 - 3.68 (2H, m), 2.62 (3H, s), 1.29 - 2.07 (9H, m). LCMS (M+H)+ m z 506 (t = 1.86 min.).
Example 485
Figure imgf000187_0002
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(l-piperidin-l-yl- methanoyl)-lH-benzoimidazol-2-yl]-lH-pyridin-2-one: *H NMR (300 MHz, CD3OD) δ 7.53 (IH, s), 7.46 (IH, s), 7.18 - 7.41 (4H, m), 7.07 (IH, s), 6.23 (IH, d, 7 = 7.6 Hz), 4.98 (IH, dd, 7 = 4.8, 6.6 Hz), 3.60 - 3.74 (5H, m), 3.18 - 3.24 (IH, m), 2.60 (3H, m), 1.43 - 1.7 (6H, m). LCMS (M+H)+ m/z 506 (t = 1.78 min.).
Example 486
Figure imgf000187_0003
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carboxylic acid cyclohexylamide: 1H NMR (300 MHz, CD3OD) δ 7.88 (IH, s), 7.54 (IH, s), 7.52 (IH, s), 7.16 - 7.40 (4H, m), 6.22 (IH, 7 = 7.6 Hz), 4.97 (IH, dd, 7 = 4.7, 6.9 Hz), 3.89 (H, m), 3.69 (IH, dd, 7 = 4.7, 13.5 Hz), 3.61 (IH, dd, 7 = 6.9, 13.5 Hz), 2.61 (3H, s), 1.19 - 2.00 (10H, m). LCMS (M+H)+ m/z 520 (t = 1.93 min.).
Example 487
Figure imgf000188_0001
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{4-methyl-6-[l-(4-methyl- piperidin-l-yl)-methanoyl]-lH-benzimidazol-2-yl}-lH-pyridin-2-one: "H NMR (300 MHz, CD3OD) δ 7.51 (IH, s), 7.49 (IH, s), 7.21 - 7.39 (4H, m), 7.14 (IH, s), 6.24 (IH, d, 7 = 7.6 Hz), 4.96 (IH, dd, 7 = 4.7, 7.0 Hz), 3.68 (IH, dd, 7 = 4.7, 13. 6 Hz), 3.60 (IH, dd, 7= 7.0, 13.6 Hz), 2.62 (3H, s), 1.93 - 1.72 (9H, m), 1.00 (3H, d, 7 = 6.3 Hz). LCMS (M+H)+ m/z 520 (t = 1.97 min.).
Example 488
Figure imgf000188_0002
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[6-(4,5-dihydro-thiazol-2-yl)-4- methyl-lH-benzimidazol-2-yl]-lH-pyridin-2-one: To a solution of (±)-2-{4-[2-(3- chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro-pyridin-3-yl } -7-methyl- 3H-benzimidazole-5-carbonitrile (100 mg, 0.24 mmol) in methanol (20 mL) was added 2-aminoethanethiol hydrochloride (41 mg, 0.36 mmol) and triethylamine (0.1 mL, excess). The reaction mixture was heated to reflux for 14 h and cooled to room temperature. After concentration, the residue was purified by prep. HPLC to yield the title compound (76 mg, 66%) as a yellow solid. 1H NMR (300 MHz, CD3OD) δ 8.10 (IH, s), 7.71 (IH, s), 7.66 (IH, s), 7.25 - 7.53 (4H, m), 6.33 (IH, d, 7 = 7.6 Hz), 5.01 (IH, dd, 7 = 4.2, 7.0 Hz), 4.58 (2H, t, 7 = 8.6 Hz), 3.91 (2H, t, 7 = 8.6 Hz), 3.73 (IH, dd, 7 = 4.2, 13.7 Hz), 3.63 (IH, dd, 7 = 7.0, 13.7 Hz), 2.64 (3H, s). LCMS (M+H)+ m/z 480 (t = 1.70 min.).
Example 489
Figure imgf000189_0001
(±)-2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3- yl}-7-methyl-3H-benzimidazole-5-carbaldehyde: To a suspension of (±)-2-{4-[2-(3- chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3-yl}-7-methyl- 3H-benzimidazole-5-carbonitrile (76 mg, 0.18 mmol) in toluene (anhydrous, 20 mL) was added diisobutylaluminium (1.4 M toluene solution) (0.65 mL, 0.97 mmol) at - 78°C under nitrogen. The mixture was stirred at -78°C for 6 h. Ethyl acetate (1 mL) was then added followed by water (0.5 mL). The mixture was stirred at room temperature for 20 min. The mixture was then passed through a pad of celite and the filtrate was concentrated. The crude product was purified by prep. HPLC to yield the title compound (4 mg, 2.5%) as a brown solid. 1H NMR (300 MHz, CD OD) δ 7.65 (IH, s), 7.57 (IH, s), 7.24 - 7.50 (5H, m), 6.26 (IH, d, 7 = 7.6 Hz), 5.49 (IH, s), 4.96 (IH, m), 3.53 - 3.79 (2H, m), 2.62 (3H, s). LCMS (M+H)+ m/z 423 (t = 1.79 min.). Example 490
Figure imgf000190_0001
(±)-4- [2-(3 -Chloro-phenyl)-2-hydroxy-ethyl amino] -3 - [6-( 1 -hydroxy- 1 -methyl-ethyl)- 4-methyl-lH-benzimidazol-2-yl]-lH-pyridin-2-one: To a solution of (+)-2-{4-[2-(3- chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro-pyridin-3-yl } -7-methyl- 3H-benzimidazole-5-carboxylic acid ethyl ester (40 mg, 0.08 mmol) in THF (5 mL) was added methyllithium (1.4 M THF solution, 0.57 mL, 0.8 mmol) at -78°C under nitrogen. The reaction mixture was gradually warmed to room temperature overnight. After quenching with water, the mixture was partitioned between ethyl acetate and water. The organic layers were washed with brine, dried over Na2SO4, and concentrated in vacuo. The residue was purified by prep HPLC to yield the title compound (13 mg, 36%) as a colorless oil. 1H NMR (300 MHz, CD3OD) δ 7.66 (IH, s), 7.47 (IH, s), 7.26 - 7.41 (5H, m), 6.26 (IH, d, 7 = 7.6 Hz), 4.92 (IH, m), 3.54 - 3.61 (2H, m), 2.62 (3H, s), 1.60 (6H, s). LCMS (M+H)+ m/z 453 (t = 1.46 min.).
Example 491
Figure imgf000190_0002
(±)-3-(6-Aminomethyl-4-methyl-lH-benzimidazol-2-yl)-4-[2-(3-chloro-phenyl)-2- hydroxy-ethylamino]-lH-pyridin-2-one: To a solution of (±)-2-{4-[2-(3-Chloro- phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro-pyridin-3-yl } -7-methyl-3H- benzimidazole-5-carboxylic acid amide (20 mg, 0.046 mmol) in THF (1 mL) was added borane-tetrahydrofuran complex (1 M solution) (0.45 mL, 0.45 mmol). The reaction mixture was stirred at room temperature for 10 h and quenched with acetic acid (2 drops). After removal of most solvent, the residue was extracted with EtOAc, washed with brine, dried over Na2SO4. After concentration, the crude product was purified by prep. HPLC to yield the title compound (11.5 mg, 60%). 1H NMR (400 MHz, CD3OD) δ 7.53 (IH, s), 7.50 (IH, s), 7.39 (IH, d, 7 = 7.5 Hz), 7.24 - 7.32 (3H, m), 7.13 (IH, s), 6.26 (IH, d, 7 = 7.5 Hz), 4.99 (IH, dd, 7 = 4.8, 6.8 Hz), 4.19 (2H, s), 3.72 (IH, dd, 7 = 4.8, 13.6 Hz), 3.64 (IH, dd, 7 = 6.8, 13.6 Hz), 2.62 (3H, s). LCMS (M+H)+ m/z 424 (t = 2.10 min.).
Example 492
Figure imgf000191_0001
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(6-hydroxymethyl-4-methyl-lH- benzimidazol-2-yl)-lH-pyridin-2-one: To a solution of (±)-2-{4-[2-(3-Chloro-phenyl)-2- hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3-yl}- -3H-benzimidazole-5-carboxylic acid ethyl ester (25 mg, 0,054 mmol) in methanol (2 mL) and acetic acid (1 mL) was added NaBIL; (10 mg, 0.27 mmol) at -20°C. The reaction mixture was stirred at -20°C for 30 min. and quenched with tsøpropanol (5 drops). After removal of most solvent, the residue was extracted with EtOAc, washed with water and brine, dried over Na2SO4. After concentration, the crude product was purified by prep. HPLC to yield the title compound (18 mg, 62%). Η NMR (300 MHz, CD3OD) δ 7.50 (IH, s), 7.48 (IH, s), 7.25 - 7.38 (4H, m), 7.22 (IH, s), 6.25 (IH, d, 7 = 7.6 Hz), 4.89 - 4.94 (IH, m), 4.76 (2H, s), 3.51 - 3.62 (2H, m), 2.62 (3H, s). LCMS (M+H)+ m z 425 (t = 1.64 min.).
Figure imgf000192_0001
Figure imgf000193_0001
General Procedure for Suzuki couplings, Example 505 -509 (Scheme N, 18)
Example 505
Figure imgf000194_0001
(S)-4-( 1 -Benzyl-2-hydroxy-ethylamino)-3-(4-methyl-6-phenyl- 1 H-benzimidazol-2- yl)-lH-pyridin-2-one: To a solution of (S)-4-(l-benzyl-2-trityloxy-ethylamino)-3-[6- bromo-4-methyl-lH-benzimidazole-2-yl]-lH-pyridin-2-one (50 mg, 0.072 mmol), phenylboronic acid (13 mg, 0.11 mmol), and 2 M K2CO3 (0.108 mL, 0.22 mmol) in THF (5 mL) was added Pd(PPh3)4 (8.3 mg, 0.007 mmol). The mixture was heated to reflux 14 h. Upon cooling, the reaction mixture was diluted with CH2C12, washed with saturated ΝaHCO3, dried over Na2SO , and concentrated in vacuo. The residue was used for the next step without purification. LCMS (M+H)+ m/z 693 (t = 2.82 min.). The crude product was treated with 4 N HCl dioxane solution (5 mL) at room temperature for 6 h. After concentration in vacuo, the residue was purified by prep. HPLC to yield the title compound (17 mg, 34%) as a white solid. 1H NMR (300 MHz, CD3OD) δ 7.13 - 7. 67 (13H, m), 6.15 (IH, d, 7 = 7.4 Hz), 3.99 - 4.11 (IH, m), 3.74 - 3.77 (2H, m), 3.16 (IH, dd, 7 = 5.4, 13.6 Hz), 2.97 (IH, dd, 7 = 7.8, 13.6 Hz), 2.69 (3H, s). LCMS (M+H)+ m/z 451 (t = 2.04 min.).
Example 506
Figure imgf000194_0002
(S)-4-( 1 -Benzyl-2-hydroxy-ethylamino)-3-(4-methyl- 1 H-benzimidazol-2-yl)- 1 H- pyridin-2-one: 'H NMR (300 MHz, CD3OD) δ 6.98 - 7.46 (9H, m), 6.15 (IH, d, 7 = 7.5 Hz), 3.99 - 4.04 (IH, m), 3.75 (2H, d, 7 = 5.1 Hz), 3.16 (IH, dd, 7= 5.1, 13.6 Hz), 2.95 (IH, dd, 7 = 8.1, 13.6 Hz), 2.61 (3H, s). LCMS (M+H)+ m z 375 (t = 1.72 min.).
Example 507
Figure imgf000195_0001
(S)-4-(l-Benzyl-2-hydroxy-ethylamino)-3-[6-(2-methoxy-phenyl)-4-methyl-lH- benzimidazol-2-yl]-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 6.99 - 7.59 (12H, m), 6.16 (IH, d, 7 =7.5 Hz), 4.04 (IH, m), 3.81 (3H, s), 3.76 (2H, d, 7 = 4.1 Hz), 2.96 - 3.20 (2H, m), 2.66 (3H, s). LCMS (M+H)+ m/z 481 (t = 2.00 min.).
Example 508
Figure imgf000195_0002
(S)-4-(l-Benzyl-2-hydroxy-ethylamino)-3-[6-(4-fluoro-phenyl)-4-methyl-lH- benzimidazol-2-yl]-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.10 - 7.66 (12H, m), 6.13 (IH, d, 7= 7.6 Hz), 3.99 - 4.04 (IH, m), 3.76 (2H, d, 7= 5.0 Hz), 3.16 (IH, dd, 7 = 5.0, 13.6 Hz), 2.96 (IH, dd, 7 = 8.1, 13.6 Hz), 2.65 (3H, s). LCMS (M+H)+ m/z 469 (t = 2.07 min.). Example 509
Figure imgf000196_0001
(S)-4-(l-Benzyl-2-hydroxy-ethylamino)-3-[6-(4-methoxy-phenyl)-4-methyl-lH- benzimidazol-2-yl]-lH-pyridin-2-one: !H NMR (300 MHz, CD3OD) δ 6.97 - 7.65 (12H, m), 6.14 (IH, d, 7 = 7.6 Hz), 4.00 - 4.04 (IH, m), 3.83 (3H, s), 3.76 (2H, d, 7 ■ 5.0 Hz), 2.95 - 3.19 (2H, m), 2.67 (3H, s). LCMS (M+H)+ m/z 481 (t = 2.01 min.).
General Procedure for Buchwald couplings, Examples 510 - 516 Scheme V, 19
Example 510
Figure imgf000196_0002
(S)-4-(l-Benzyl-2-hydroxy-ethylamino)-3-(4-methyl-6-moφholin-4-yl-lH- benzimidazol-2-yl)-lH-pyridin-2-one: A mixture of (S)-4-(l -benzyl -2-trityloxy- ethylamino)-3 - [6-bromo-4-methyl- 1 H-benzimidazol-2-yl] - 1 H-pyridin-2-one (150 mg, 0.216 mmol), moφholine (28.2 mg, 0.324 mmol), palladium acetate (2.4 mg, 0.01 mmol), tri-tert-buylphosphine (4.4 mg. 0.02 mmol), and sodium tert-butoxide (104 mg, 1.08 mmol) in toluene (5 mL) was heated to 100°C for 14 h under nitrogen. The reaction mixture was cooled to room temperature and diluted with EtOAc. After extraction, the combined organic layers were washed with water, brine, dried over Na2SO4. Concentration gave a brownish residue, which was treated with 4 N HCl dioxane solution (3 mL) at room temperature for 6 h. After removal of the solvent, the residue was purified by prep. HPLC to yield the title compound (18 mg, 18%). 1H NMR (400 MHz, CD3OD) δ 7.59 (IH, s), 7.12 - 7.28 (7H, m), 6.12 (IH, d, 7= 7.6 Hz), 4.01 - 4.08 (5H, m), 3.76 (IH, dd, 7 = 4.8, 11.1 Hz), 3.71 (IH, dd, 7= 5.2, 11.1 Hz), 3.32 - 3.10 (4H, m), 3.09 (IH, dd, 7 = 5.6, 13.7 Hz), 2.92 (IH, dd, 7= 8.0, 13.7 Hz), 2.65 (3H, s). LCMS (M+H)+ m/z 460 (t = 1.30 min.).
Example 511
Figure imgf000197_0001
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-lH-benzimidazol-2- yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.17 - 7.64 (8H, m), 6.23 (IH, d, 7= 10.2 Hz), 4.94 (IH, m), 3.61 (IH, dd, 7 = 4.8, 13.8 Hz), 3.54 (IH, dd, 7 = 7.4, 13.8 Hz), 2.61 (3H, s). LCMS (M+H)+ m/z 395 (t = 1.65 min.).
Example 512
Figure imgf000197_0002
(±)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-pyridin-3-yl-lH- benzimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 9.14 (IH, s), 8.88 (IH, d, 7= 8.2 Hz), 8.72 (IH, d, 7 = 5.4 Hz), 8.08 (IH, dd, 7= 5.8, 8.2 Hz), 7.82 (IH, s), 7.56 (IH, s), 7.24 - 7.45 (5H, m), 6.26 (IH, d, 7 = 7.6 Hz), 5.02 (IH, dd, 7 = 5.0, 6.5 Hz), 3.77 (IH, dd, 7 = 5.0, 13.5 Hz), 3.68 (IH, dd, 7= 6.5, 13.5 Hz), 2.68 (3H, s). LCMS (M+H)+ m/z 472 (t = 1.66 min.).
Figure imgf000198_0002
The following examples 517-519 were prepared according to Schemes NH and EL
Example 517
Figure imgf000198_0001
(S )-4- [2-(3 -Chloro-phenyl)-2-hydroxy-ethylamino] -3 -(4-methyl-6-piperazin- 1 -yl- 1 H- benzoimidazol-2-yl)-lH-pyridin-2-one: 1H ΝMR (500 MHz, CD3OD) δ 7.54 (brs, IH), 7.40 - 7.20 (m, 4H), 7.03 (brs, IH), 6.84 (brs, IH), 6.25 (d, IH, J = 7.60 Hz), 5.01 - 4.91 (m, IH), 3.73 (dd, IH), 3.65 (dd, IH), 3.45 - 3.25 (m, 8H), 2.56 (s, 3H); LCMS (M+H)+ m/z 479,481.
Example 518
Figure imgf000199_0001
(±)-4-[2-(3-Chloro-4-methylsulfanyl-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6- piperazin-l-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one: *H NMR (500 MHz, CD3OD) δ 7.47 (brs, IH), 7.38 - 7.29 (m, 2H), 7.19 (d, IH, J = 8.3 Hz), 7.03 (brs, IH), 6.98 (brs, IH), 6.26 (d, IH, J = 7.7 Hz), 4.90 - 4.81 (m, IH), 3.65 - 3.35 (m, 10H), 2.56 (s, 3H), 2.42 (s, 3H); LCMS (M+H)+ m/z 525, 527.
Example 519
Figure imgf000199_0002
(S)-4-[2-(3-Chloro-4-fluoro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-piperazin- l-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (500 MHz, CD3OD) δ 7.61 (dd, IH, J = 2.1, 7.2 Hz), 7.40 (ddd, IH), 7.28 (d, IH, J = 7.5 Hz), 7.17 (dd, IH, J = 8.9, 8.8 Hz), 7.02 (brs, IH), 6.87 (brs, IH), 6.25 (d, IH, J = 7.6 Hz), 4.99 - 4.90 (m, IH), 3.73 - 3.60 (m, 2H), 3.45 - 3.30 (m, 8H), 2.54 (s, 3H); LCMS (M+H)+ m/z 497, 499. The following examples (520-522) were prepared according to Scheme NH and HI and illustrate the alkylation of a piperazine derivative
Example 520 (General procedure for examples 520-522)
Figure imgf000200_0001
(S)-3-[4-(2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl } -7-methyl-3H-benzoimidazol-5-yl)-piperazin- l-yl]-propionitrile: To a stirred solution of 4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6- piperazin-l-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one (17 mg, 0.021 mmol) in anhydrous methanol (1 mL) was added Hunigs base (36 μL). The resulting solution was cooled to 0 °C and acrylonitrile (5 μL) was added in portions until the reaction was complete as judged by LCMS. The reaction was warmed to room temperature and the solvent evaporated in vacuo. The resulting residue was purified on reverse phase preparative HPLC using a methanol / water 10.1% trifluoroacetic acid gradient. The fractions were evaporated to give the title compound as a trifluoroacetic acid salt (12.1 mg): 1H ΝMR (500 MHz, CD3OD) δ 7.49 (brs, IH), 7.39 - 7.23 (m, 4H), 7.15 (brs, IH), 7.06 (brs, IH), 6.25 (d, IH, J = 7.7 Hz), 4.97 - 4.88 (m, IH), 3.70 - 3.40 (m, 12 H), 3.06 (t, 2H, J = 7.0 Hz), 2.59 (s, 3H); LCMS (M+H)+ m/z 532,534. The tiifluoroaceticacid salt of the pure title compound was dissolved in methanol and applied to a Narian Mega Bond-Elute SCX cartridge. Elution with methanol followed by 2.0 M ΝH3 / MeOH gave the free base. This material was suspended in MeOH and 1.00 N aqueous HCl (2 equiv.) was added. The resulting solution was filtered through a 45 μm filter and evaporated to give the bis HCl salt of the title compound: 1H NMR (300 MHz, CD3OD) δ 7.50 - 7.25 (m, 5H), 7.16 (brs, IH), 7.13 (brs, IH), 6.27 (d, IH, J = 7.7 Hz), 4.95 - 4.87 (m, IH), 3.70 - 3.40 (m, 12H), 3.13 (t, 2H, J = 7.0 Hz), 2.62 (s, 3H); LCMS (M+H)+ m/z 532,534. Example 521
Figure imgf000201_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-methanesulfonyl- ethyl)-piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one: *H NMR (500 MHz, CD3OD) δ 7.53 (brs, IH), 7.41 - 7.20 (m, 4H), 7.13 (brs, IH), 6.92 (brs, IH), 6.24 (d, IH, J = 7.6 Hz), 5.00 - 4.92 (m, IH), 3.80 - 3.25 (m, 14H), 3.13 (s, 3H), 2.57 (s, 3H); LCMS (M+H)+ m/z 585, 587.
Example 522
Figure imgf000201_0002
(S)-3-[4-(2-{4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2- dihydro-pyridin-3-yl}-7-methyl-3H-benzoimidazol-5-yl)-piperazin-l-yl]- propionitrile: *H NMR (500 MHz, CD3OD) δ 7.59 (brs, IH), 7.41 (d, IH, J = 7.5 Hz), 7.34 (dd, IH, J = 2.0, 8.5 Hz), 7.17 (brs, IH), 7.12 (brs, IH), 7.00 (d, IH, J = 8.5 Hz), 6.25 (d, IH, J = 7.5 Hz), 4.85 - 4.76 (m, IH), 3.85 (s, 3H), 3.80 - 3.30 (m, 12H), 3.16 (t, 2H, J = 7.0 Hz), 2.55 (s, 3H); LCMS (M+H)+ m/z 606, 608. The following examples (523-528) were prepared according to Scheme NH and IH and illustrate the carbamoylation of a piperazine derivative
Example 523 (General procedure for examples 523-528)
Figure imgf000202_0001
(S)-4-(2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin- 3-yl}-7-methyl-3H-benzoimidazol-5-yl)-piperazine-l-carboxylic acid 2-fluoro-ethyl ester: To a stirred solution of 4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4- methyl-6-piperazin-l-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one (~ 2 TFA salt, 80 mg, -0.1 mmol) in methanol (2 mL) at 0 °C was added N,N-diisopropylethylamine (170 μL) and 2-fluoroethyl chloroformate (37 mg). The cooling bath was removed and the solution was stirred at room temperature for 30 minutes, after which LC/MS analysis showed the reaction to be complete. The reaction mixture was then purified on reverse phase preparative HPLC using a methanol / water / 0.1% trifluoroacetic acid gradient. The fractions were evaporated to give the title compound as a trifluoroacetic acid salt, which was dissolved in methanol and applied to a Varian Mega Bond-Elute SCX cartridge. Elution with methanol followed by 2.0 M NH3 / MeOH gave the free base (46.2 mg). This material was suspended in MeOH and 1.00 N aqueous HCl (1 equiv.) was added. The resulting solution was filtered through a 45 μm filter and evaporated to give the mono HCl salt of the title compound (46 mg): 1H NMR (500 MHz, CD3OD) δ 7.50 (brs, IH), 7.45 - 7.20 (m, 6H), 6.26 (d, IH, J = 7.7 Hz), 4.98 - 4.91 (m, IH), 4.64 (dm, 2H, J = 47.9 Hz), 4.39 (dm, 2H, J = 29 Hz), 3.95 - 3.50 (m, 10H), 2.63 (s, 3H); . LCMS (M+H)+ m/z 569,571. Example 524
Figure imgf000203_0001
(S)-4-(2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin- 3-yl } -7-methyl-3H-benzoimidazol-5-yl)-piperazine- 1 -carboxylic acid 2-methoxy- ethyl ester: 1H NMR (500 MHz, CD3OD) δ 7.53 - 7.22 (m, 7H), 6.26 (d, IH, J = 7.6 Hz), 4.96 (dd, IH, J = 7.0, 4.6 Hz), 4.31 - 4.27 (m, 2H), 4.05 - 3.55 (m, 12H), 3.39 (s, 3H), 2.64 (s, 3H); LCMS (M+H)+ m/z 581, 583.
Example 525
Figure imgf000203_0002
(S)-4-(2- { 4-[2-(3-Chloro-ρhenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro-pyridin- 3-yl}-7-methyl-3H-benzoimidazol-5-yl)-piperazine-l -carboxylic acid tert-butyl ester: 1H NMR (500 MHz, CD3OD) δ 7.51 (brs, IH), 7.40 - 7.22 (m, 6H), 6.26 (d, IH, J = 7.7 Hz), 4.96 - 4.90 (m, IH), 3.90 - 3.30 (m, 10H), 2.64 (s, 3H), 1.51 (s, 9H); LCMS (M+H)+ m/z 579, 581. Example 526
Figure imgf000204_0001
(S)-4-(2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin- 3-yl } -7-methyl-3H-benzoimidazol-5-yl)-piperazine- 1 -carboxylic acid prop-2-ynyl ester: 1H NMR (500 MHz, CD3OD) δ 7.50 (brs, IH), 7.43 - 7.20 (m, 5H), 7.19 (brs, IH), 6.25 (d, IH, J = 7.6 Hz), 4.98 - 4.90 (m, IH), 4.78 (d, 2H, J = 2.5 Hz), 3.95 - 3.30 (m, 10H), 2.97 (t, IH, J = 2.5 Hz), 2.62 (s, 3H); LCMS (M+H)+ m/z 561, 563.
Example 527
Figure imgf000204_0002
(S)-4-(2-{4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2- dihydro-pyridin-3-yl}-7-methyl-3H-benzoimidazol-5-yl)-piperazine-l-carboxylic acid tert-butyl ester: 1H NMR (300 MHz, CD3OD) δ 7.65 (brs, IH), 7.48 - 7.28 (m, 3H), 7.25 (brs, IH), 6.97 (d, IH, J = 8.5 Hz), 6.25 (d, IH, J = 7.7 Hz), 4.94 - 4.86 (m, IH), 3.90 - 3.45 (m, 10H), 3.82 (s, 3H), 2.61 (s, 3H), 1.52 (s, 9H); LCMS (M+H)+ m/z 653, 655. Example 528
Figure imgf000205_0001
(S)-4-(2-{4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl}-7-methyl-3H-benzimidazol-5-yl)-piperazine-l-carboxylic acid ethyl ester: 1H NMR (400 MHz, CD3OD) δ 7.80 (IH, s), 7.63 (IH, narrow d, 7 = 1.8 Hz), 7.52 (IH, s), 7.42 (IH, d, 7= 7.5 Hz), 7.37 (IH, dd, 7 = 1.8, 8.4 Hz), 7.00 (IH, d, 7= 8.4 Hz), 6.31 (IH, d, 7 = 7.5 Hz), 4.89 (IH, m), 4.22 (2H, q, 7 = 7.1 Hz), 3.98 (4H, br s), 3.84 (3H, s), 3.70 - 3.72 (4H, m), 3.59 - 3.60 (2H, m), 2.67 (3H, s), 1.31 (3H, t, 7 = 7.1 Hz). LCMS (M+H)+ m/z 625 (t = 1.45 min.).
The following examples (529-540) were prepared according to Scheme Nil and HI and illustrate an alternative method of alkylation of a piperazine derivative
Example 529 (General procedure for examples 529-540)
Figure imgf000205_0002
(S)-4-[2-(3-Chloro-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(3-fluoro- propyl)-piperazin-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: To a stirred solution of 4-[2-(3-Chloro-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4- methyl-6-piperazin-l-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one (100 mg, 0.162 mmol) in 1,4-dioxane (4.0 mL), ethanol (0.8 mL), methanol (0.8 mL) was added N,N- diisopropylethylamine (0.30 mL) and l-bromo-3-fluoropropane (85 μl). The reaction was heated at 80 °C for 12 h. The reaction mixture was then purified on reverse phase preparative HPLC using a methanol / water / 0.1% trifluoroacetic acid gradient. The fractions were evaporated to give the title compound as a trifluoroacetic acid salt, which was dissolved in methanol and applied to a Varian Mega Bond-Elute SCX cartridge. Elution with methanol followed by 2.0 M NH3 / MeOH gave the free base (39.3 mg). This material was suspended in MeOH and 1.00 N aqueous HCl (2 equiv.) was added. The resulting solution was filtered through a 45 μm filter and evaporated to give the bis HCl salt of the title compound (43.5 mg): 1H NMR (400 MHz,
CD3OD) δ 7.43 - 7.37 (m, 2H), 7.28 (dd, IH, J = 2.0, 8.6 Hz), 7.16 (brs, IH), 7.08 (d, IH, J = 1.7 Hz), 7.01 (d, IH, J = 8.5 Hz), 6.24 (d, IH, J = 7.7 Hz), 4.84 - 4.78 (m, IH), 4.60 (dt, 2H, J = 5.7, 47.1 Hz), 3.95 - 3.88 (m, 2H), 3.83 (s, 3H), 3.78 - 3.71 (m, 2H), 3.53 - 3.14 (m, 8H), 2.59 (s, 3H), 2.31 - 2.16 (m, 2H); LCMS (M+H)+ m/z 569, 571.
Example 530
Figure imgf000206_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-fluoro-ethyl)- piperazin-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.45 - 7.20 (m, 5H), 7.16 (brs, IH), 7.09 (brs, IH), 6.25 (d, IH, J = 7.6 Hz), 5.00 - 4.92 (m, IH), 4.92 - 4.78 (m, 2H), 3.98 - 3.15 (m, 12H), 2.60 (s, 3H); LCMS (M+H)+ m/z 525, 527. Example 531
Figure imgf000207_0001
(S)-4-[2-(3-Chloro-4-fluoro-ρhenyl)-2-hydroxy-ethylamino]-3- { 6-[4-(3-fluoro- propyl)-piperazin- 1 -yl] -4-methyl- lH-benzoimidazol-2-yl } - 1 H-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.52 (dd, IH, J = 2.1, 7.2 Hz), 7.40 (d, IH, J = 7.6 Hz), 7.35 - 7.12 (m, 3H), 7.08 (d, IH, J = 1.7Hz), 6.25 (d, IH, J = 7.7Hz), 4.90 - 4.82 (m, IH), 4.60 (dt, 2H, J = 5.4, 47.1 Hz), 3.96 - 3.10 (m, 12H), 2.60 (s, 3H), 2.28 - 2.13 (m, 2H); LCMS (M+H)+ m/z 557, 559.
Example 532
Figure imgf000207_0002
(S)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(3-fluoro- propyl)-piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.58 (d, IH, J = 2.0 Hz), 7.43 (d, IH, J = 7.6 Hz), 7.34 (dd, IH, J = 2.0, 8.5 Hz), 7.19 (brs, IH), 7.11 (brs, IH), 7.00 (d, IH, J = 8.5 Hz), 6.26 (d, IH, J = 7.7 Hz), 5.90 - 4.82 (m, IH), 4.61 (dt, 2H, J = 5.4, 47.1 Hz), 3.95 - 3.12 (m, 12H), 3.85 (s, 3H), 2.62 (s, 3H), 2.30 - 2.14 (m, 2H); LCMS (M+H)+ m/z 613, 615. Example 533
Figure imgf000208_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{4-methyI-6-[4-(3,3,3-trifluoro- propyl)-piperazin-l-yl]-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.50 - 7.10 (m, 7H), 6.26 (d, IH, J = 7.5 Hz), 4.96 - 4.88 (m, IH), 4.00 - 3.15 (m, 12H), 3.00 - 2.82 (m, 2H), 2.61 (s, 3H); LCMS (M+H)+ m/z 575, 577.
Example 534
Figure imgf000208_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(3-fluoro-propyl)- piperazin-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: !H NMR (300 MHz, CD3OD) δ 7.46 - 7.25 (m, 5H), 7.19 (brs, IH), 7.11 (brs, IH), 6.27 (d, IH, J = 7.7 Hz), 4.95 - 4.86 (m, IH), 4.62 (dt, 2H, J = 5.4, 47.1 Hz), 3.98 - 3.15 (m, 12H), 2.62 (s, 3H), 2.35 - 2.12 (m, 2H); LCMS (M+H)+ m/z 539, 541. Example 535
Figure imgf000209_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{4-methyl-6-[4-(3,4,4-trifluoro- but-3-enyl)-piperazin- 1 -yl]- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one: LCMS (M+H)+ m/z 587, 589.
Example 536
Figure imgf000209_0002
4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-{6-[4-(3-fluoro-2-hydroxy- propyl)-piperazin-l-yl]-4-methyl-lH-benzoimidazol-2-yl } -lH-pyridin-2-one: LCMS (M+H)+ m/z 555, 557.
Example 537
Figure imgf000209_0003
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-hydroxy-2-methyl- propyl)-piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one : " H NMR (400 MHz, CD3OD) δ 7.44 - 7.22 (m, 5H), 7.15 (brs, IH), 7.08 (brs, IH), 6.25 (d, IH, J = 7.6 Hz), 4.88 - 4.80 (m, IH), 3.86 - 3.81 (m, 4H), 3.55 - 3.32 (m, 6H), 3.28 (s, 2H), 2.60 (s, 3H), 1.38 (s, 6H); LCMS (M+H)+ m/z 551, 553.
Example 538
HO^
Figure imgf000210_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-hydroxy-ethyl)- piperazin- 1 -yl]-4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one: ' HNMR (400 MHz, CD3OD) δ 7.50 (IH, s), 7.25 - 7.35 (4H, m), 7.04 (IH, s), 6.97 (IH, s), 6.22 (IH, d, 7 = 6.8 Hz), 4.93 - 4.95 (IH, m), 3.21 - 3.96 (14H, m), 2.57 (3H, s). LCMS (M+H)+ m/z 523 (t = 1.11 min.).
Example 539
Figure imgf000210_0002
(S)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-hydroxy- ethyl)-piperazin- 1 -yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one : * H NMR (400 MHz, CD3OD) δ 7.60 (IH, narrow d, 7 = 2.0 Hz), 7.40 (IH, d, 7 = 7.6 Hz), 7.35 (IH, dd, 7 = 2.0, 8.4 Hz), 7.13 (IH, s), 7.09 (IH, s), 6.99 (IH, d, 7 = 8.4 Hz), 6.25 (IH, d, 7 = 7.6 Hz), 4.82 - 4.87 (IH, m), 3.76 - 3.97 (6H, m), 3.84 (3H, s), 3.24 - 3.52 (8H, m), 2.60 (3H, s). LCMS (M+H)+ m z 597 (t = 1.09 min.).
Example 540
Figure imgf000211_0001
(S)-[4-(2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin- 3-yl}-7-methyl-3H-benzoimidazol-5-yl)-piperazin-l-yl]-acetonitrile: 1H NMR (400 MHz, CD3OD) δ 7.50 - 7.25 (m, 7H), 6.28 (d, IH, J = 8.0 Hz), 4.95 - 4.88 (m, IH), 4.32 (s, 2H), 3.75 - 3.62 (m, 4H), 3.60 - 3.35 (m, 6H), 2.63 (s, 3H); LCMS (M+H)+ m/z 518, 520.
The following examples (541-553) were prepared according to Scheme NH and HI and illustrate the acylation of a piperazine derivative
Example 541 (General procedure for examples 541-553)
Figure imgf000211_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(4-fluoro-butyryl)- piperazin-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: To a stirred solution of 4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-piperazin- l-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one (70 mg, 0.136 mmol) in anhydrous N,N-dimethylformamide (750 μl) was added l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (112 mg, 0.584 mmol), 1-hydroxybenzotriazole hydrate (59 mg, 0.438 mmol), N-methylmoφholine (0.048 mL, 0.438 mmol) and 4- fluorobutyric acid (31 mg, 0.291 mmol; see O'Hagan,D., J. Fluorine Chem., 43, (1989), 371-377) and the mixture heated to 80 °C for 3 h. The reaction mixture was then purified on reverse phase preparative HPLC using a methanol / water / 0.1% trifluoroacetic acid gradient. The fractions were evaporated to give the title compound as a trifluoroacetic acid salt, which was dissolved in methanol and applied to a Varian Mega Bond-Elute SCX cartridge. Elution with methanol followed by 2.0 M NH3 / MeOH gave the free base (35.9 mg). This material was suspended in MeOH and 1.00 N aqueous HCl (2 equiv.) was added. The resulting solution was filtered through a 45 μm filter and evaporated to give the bis HCl salt of the title compound (37.6 mg): 1H NMR (400 MHz, CD3OD) δ. 7.61 (brs, IH), 7.47 (brs, IH), 7.44 - 7.20 (m, 5H), 6.27 (d, IH, J = 7.6 Hz), 4.92 (dd, IH, J = 4.4, 7.3 Hz), 4.50 (dt, 2H, J = 5,9, 47.3 Hz), 4.05 - 3.40 (m, 10H), 2.64 (s, 3H), 2.63 (t, 2H, J = 7.6 Hz), 2.09 - 1.95 (m, 2H); LCMS (M+H)+ m/z 567,569.
Example 542
Figure imgf000212_0001
(S)-4-[2-(3-Chloro-ρhenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2,2-difluoro-acetyl)- piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one : LCMS (M+H)+ m/z 557, 559. Example 543
Figure imgf000213_0001
(S)-4- [2-(3 -Chloro-phenyl)-2-hydroxy-ethylamino] -3 - { 6- [4-(2-methanesulfonyl- acetyl)-piperazin- 1 -yl]-4-methyl- lH-benzoimidazol-2-yl } - lH-pyridin-2-one: LCMS (M+H)+ m/z 599, 601.
Example 544
Figure imgf000213_0002
(S)-3 - [6-(4- Acetyl-piperazin- 1 -yl)-4-methyl- 1 H-benzoimidazol-2-yl] -4- [2-(3 -chloro- phenyl)-2-hydroxy-ethylamino]-lH-ρyridin-2-one: 1H NMR (500 MHz, CD3OD) δ 7.57 (brs, IH), 7.49 (brs, IH), 7.40 - 7.20 (m, 5H), 6.29 (d, IH, J = 7.6 Hz), 4.98 - 4.90 (m, IH), 4.02 - 3.91 (m, 4H), 3.70 - 3.50 (m, 6H), 2.66 (s, 3H), 2.21 (s, 3H); LCMS (M+H)+ m/z 521, 523.
Example 545
Figure imgf000213_0003
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6- { 4-[2-( 1 -oxo- 114- thiomoφholin-4-yl)-acetyl] -piperazin- 1 -yl } - 1 H-benzoimidazol-2-yl)- 1 H-pyridin-2- one: LCMS (M+H)+ m/z 638, 640.
Example 546
Figure imgf000214_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(6- { 4-[2-(l , 1 -dioxo- 146- thiomoφholin-4-yl)-acetyl] -piperazin- 1 -yl } -4-methyl- 1 H-benzoimidazol-2-yl)- 1 H- pyridin-2-one: LCMS (M+H)+ m z 654, 656.
Example 547
Figure imgf000214_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{4-methyl-6-[4-(2- thiomoφholin-4-yl-acetyl)-piperazin- 1 -yl] - 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2- one: LCMS (M+H)+ m/z 622, 624. Example 548
Figure imgf000215_0001
4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-{6-[4-(2-methanesulfinyl- acetyl)-piperazin-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: LCMS (M+H)+ m/z 583, 585.
Example 549
Figure imgf000215_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-methoxy-acetyl)- piperazin-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: 1H NMR (500 MHz, CD3OD) δ 7.49 (brs, IH), 7.40 - 7.18 (m, 6H), 6.26 (d, IH, J = 7.7 Hz), 4.98 - 4.90 (m, IH), 4.25 (s, 2H), 3.95 - 3.46 (m, 10H), 3.44 (s, 3H), 2.63 (s, 3H); LCMS (M+H)+ m/z 551, 553.
Example 550
Figure imgf000215_0003
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{4-methyl-6-[4-(2- methylsulf anyl-acetyl)-piperazin- 1 -yl] - 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one: Η NMR (500 MHz, CD3OD) δ 7.49 (brs, IH), 7.45 - 7.20 (m, 6H), 6.26 (d, IH, J = 7.60 Hz), 4.98 - 4.90 (m, IH), 3.97 - 3.40 (m, 12H), 2.63 (s, 3H), 2.20 (s, 3H); LCMS (M+H)+ m/z 567, 569.
Example 551
Figure imgf000216_0001
(S)-3-{6-[4-(2-Chloro-acetyl)-ρiperazin-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-4-[2- (3-chloro-phenyl)-2-hydroxy-ethylamino]-lH-pyridin-2-one: !H NMR (500 MHz, CD3OD) δ 7.49 (brs, IH), 7.38 - 7.20 (m, 5H), 7.15 (brs, IH), 6.25 (d, IH, J = 7.6 Hz), 4.98 - 4.90 (m, IH), 4.35 (s, 2H), 3.90 - 3.80 (m, 4H), 3.66 - 3.30 (m, 6H), 2.61 (s, 3H); LCMS (M+H)+ m/z 555, 557.
Example 552
Figure imgf000216_0002
(S)-4-(2- { 4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2- dihydro-pyridin-3-yl}-7-methyl-3H-benzimidazol-5-yl)-piperazine-l-carbaldehyde: 1H NMR (400 MHz, CD3OD) δ 8.18 (IH, s), 7.65 (IH, s), 7.61 (IH, narrow d, 7= 2.0 Hz), 7.46 (IH, s), 7.43 (IH, d, 7 = 7.6 Hz), 7.36 (IH, dd, 7= 2.0, 8.5 Hz), 7.00 (IH, d, 7 = 8.5 Hz), 6.32 (IH, d, 7 = 7.6 Hz), 4.86 - 4.89 (IH, m), 3.91 - 3.96 (4H, m), 3.84 (3H, s), 3.57 - 3.67 (7H, m), 2.66 (3H, s). LCMS (M+H)+ m/z 581 (t = 1.24 min.).
Example 553
Figure imgf000217_0001
(S)-4-(2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin- 3-yl }-7-methyl-3H-benzimidazol-5-yl)-piperazine-l-carbaldehyde: 1H NMR (400 MHz, CD3OD) δ 8.12 (IH, s), 7.50 (IH, s), 7.24 - 7.38 (6H, m), 6.23 (IH, d, 7= 7.6 Hz), 4.93 - 4.96 (IH, m), 3.54 - 3.79 (10H, m), 2.58 (3H, s). LCMS (M+H)+ m/z 507 (t = 1.29 min.).
The following examples (Examples 554-575) were prepared according to Scheme NE and lE
Example 554
Figure imgf000217_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin-4-yl- lH-benzoimidazol-2-yl)-l H-pyridin-2-one: To a suspension of 4-Chloro-3-(4- methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 H-pyridin-2-one and the corresponding iodo compound 4-Iodo-3-(4-methyl-6-moφholin-4-yl-lH- benzoimidazol-2-yl)-lH-pyridin-2-one (4.43 g, ~ 11 mmol) in acetonitrile (100 ml) triethylamine (7.0 ml, 50 mmol) and 2-(S)-2-(3-Chloro-phenyl)-2-hydroxy-ethylamine hydrochloride (2.55 g, 12.2 mmol) were added. The mixture was stirred at 85°C overnight. LCMS showed some starting material pyridone left. 2-(S)-2-(3-Chloro- phenyl)-2-hydroxy-ethylamine hydrochloride (0.22 g, 1.06 mmol) was added and the mixture stirred at 85°C another 24 hours. After evaporation of volatiles an aqueous solution of Cs2CO3 (200 ml, 10%) was added, the suspension sonicated for 5 minutes and stirred overnight. The product was filtered, washed with water and recrystallized from methanol - chloroform. The title compound was isolated as yellow crystals. (3.951 g, 75%). LCMS (M+H)+ m/z 480 (t = 1.31 min.). HPLC t = 4.93 min, YMC- Pack ODS-A 3.0 x 50 mm; 0 - 100% gradient over 8 min; 2.5 mL/min flow rate. 1H NMR of mono-HCl salt (500 MHz, DMSO-de) δ 13.3 (IH, broad s), 11.22 (IH, s), 10.9 (IH, broad s), 7.65 (IH, broad s), 7.60 (IH, s), 7.45 (d, 7= 7.6 Hz), 7.38-7.30 (4H, m) 6.19 (IH, d, 7 = 7.5 Hz), 4.92 (IH, t, 7 = 5.3 Hz), 4.00 (6H, broad), 3.67 (IH, m), 3.52 (5H, m), 2.58 (3H, s).
General procedure for preparation of mono- and bis- HCl salts: A solution or suspension of free base in methanol is treated with 1.00 (or 2.00 resp.) equivalent of 1.00 N aqueous HCl. If significant amounts of compound remain insoluble an equal volume of dichlorethane is added to improve solubility. The mixture is filtered and concentrated in vacuo. .Small amounts were evaporate to dryness, large-scale preparations for in vivo studies were concentrated until most of the compound crystallized, then filtered.
Example 555
Figure imgf000218_0001
(S)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6- moφholin-4-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.68 - 7.60 (m, 2H), 7.43 - 7.28 (m, 3H), 6.98 (d, IH, J = 8.50 Hz), 6.26 (d, IH, J = 7.7 Hz), 4.97 - 4.89 (m, IH), 4.18 - 4.04 (m, 4H), 3.82 (s, 3H), 3.73 - 3.55 (m, 6H), 2.63 (s, 3H); LCMS (M+H)+ m/z 554, 556.
Example 556
Figure imgf000219_0001
(S)-4-[2-(3-Chloro-4-fluoro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6- moφholin-4-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one: 1H NMR (300 MHz, CD3OD) δ 7.65 - 7.50 (m, 2H), 7.43 - 7.20 (m, 3H), 7.18 (dd, IH, J = 8.9, 8.8 Hz), 6.27 (d, IH, J = 7.6 Hz), 5.00 - 4.91 (m, IH), 4.15 - 4.02 (m, 4H), 3.75 - 3.60 (m, 6H), 2.64 (s, 3H); LCMS (M+H)+ m z 498, 500.
Example 557
Figure imgf000219_0002
(S)-4-[2-(3-Chloro-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6- moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 H-pyridin-2-one: ' H NMR (400 MHz, CD3OD) δ 7.67 (brs, IH), 7.45 (brs, IH), 7.40 - 7.28 (m, 3H), 7.00 (d, IH, J = 8.4 Hz), 6.27 (d, IH, J = 7.6 Hz), 4.95 - 4.84 (m, IH), 4.15 - 4.05 (m, 4H), 3.82 (s, 3H), 3.75 - 3.55 (m, 6H), 2.64 (s, 3H); LCMS (M+H)+ m z 510, 512.
Example 558
Figure imgf000220_0001
(S)-4-[2-(7-Bromo-2,3-dihydro-benzofuran-5-yl)-2-hydroxy-ethylamino]-3-(4- methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-lH-pyridin-2-one: *H NMR (400 MHz, CD3OD) δ 7.54 (brs, IH), 7.35 - 7.30 (m, 2H), 7.27 (brs, IH), 7.22 (brs, IH), 6.27 (d IH, J = 7.6 Hz), 4.95 - 4.87 (m, IH), 4.60 - 4.50 (m, 2H), 4.10 - 4.00 (m, 4H), 3.75 - 3.60 (m, 6H), 3.25 - 3.13 (m, 2H), 2.62 (s, 3H); LCMS (M+H)+ m/z 566, 568.
Example 559
Figure imgf000220_0002
4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-[4-methyl-6-[2(S),6(R)- dimethyl-mθφholine-4-yl]-lH-benzoimidazol-2-yl]-lH-pyridine-2-one. Η NMR (400 MHz, CD3OD) δ 7.23-7.57 (7H, - m), 6.22 (IH, d, 7= 7.6 Hz), 4.97 (IH, m), 4.06 (2H, m), 3.62-3.68 (4H, m), 3.20-3.34 (2H, m), 2.63 (3H, s), 1.30 (6H, d, 7 = 6.28 Hz), LCMS (M+H)+ m/z 508 (t = 2.12 min.) Example 560
Figure imgf000221_0001
4-[2-(3-Bromo-4-methoxy-phenyl)-2(S)-hydroxy-εthylamino]-3-[4-methyl-6- [2(S),6(R)-dimethyl-moφholine-4-yl]-lH-benzoimidazol-2-yl]-lH-pyridine-2-one.1H NMR (400 MHz, CD3OD) δ 7.73 (IH- s), 7.63 (IH, s), 7.42 (IH, s), 7.34 (2H, m), 6.96 (IH, d, 7= 8.48 Hz), 6.21 (IH, d, 7= 7.48 Hz), 4.87 (IH, m), 4.23 (2H, m), 3.57- 3.67(4H, m), 3.34 (3H, s) 3.30-3.32(2H, m), 2.60 (IH, s), 1.30 (6H, d, 7 = 6.2 Hz) LCMS (M+H)+ m/z 582 (t = 2.03 min.)
Example 561
Figure imgf000221_0002
4-[2-(3-Chloro-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[('R)-2-fluoromethyl- moφholin-4-yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one and 4-[2-(3- chloro-phenyl)-(S)-2-hyαiOxy-ethylamino]-3-{6-[('S -2-fluoromethyl-moφholin-4-yl]- 4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one : 1H NMR (400 MHz, CD3OD) δ 7.47 (IH, s), 7.23 - 7.43 (6H, m), 6.22 (IH, d, 7 = 7.2 Hz), 4.87 - 4.94 (IH, m), 4.60 (IH, d, 7= 3.4 Hz), 4.48 (IH, d, 7= 3.4 Hz), 4.04 - 4.15 (3H, m), 3.28 - 3.62 (6H, m), 2.56 (3H, s). LCMS (M+H)+ m/z 512 (t = 1.35 min.). Example 562
Figure imgf000222_0001
4-[2-(3-Bromo-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[(R)-2- fluoromethyl-moφholin-4-yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one and 4-[2-(3-bromo-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3- { 6-[(S)-2- fluoromethyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one : H NMR (400 MHz, CD3OD) δ 7.60 (IH, narrow d, J = 2.0 Hz), 7.56 (IH, s), 7.39 (IH, br s), 7.34 (IH, dd, 7 = 2.0, 8.4 Hz), 6.97 (IH, d, J 8.4 Hz), 6.27 (IH, d, 7 = 6.4 Hz), 4.86 (IH, m), 4.61 (IH, m), 4.50 (IH, m), 4.77 - 4.19 (3H, m), 3.82 (3H, s), 3.38 - 3.74 (8H, m), 2.63 (3H, s). LCMS (M+H)+ m/z 586 (t = 1.31 min.).
Example 563
Figure imgf000222_0002
4-[2-(3-Chloro-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[ R)-2- fluoromethyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one and 4-[2-(3-chloro-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3- { 6-[(S)-2- fluoromethyl-moφholin-4-yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.45 (IH, s), 7.28 - 7.30 (3H, m), 7.10 (IH, s), 6.97 (IH, d, 7= 8.0 Hz), 6.20 (IH, d, 7 = 6.4 Hz), 4.85 (IH, m), 4.58 (IH, br s), 4.46 (IH, br s), 3.93 - 4.10 (3H, m), 3.80 (3H, s), 3.59 (4H, m), 3.07 - 3.29 (2H, m), 2.55 (3H, s). LCMS (M+H)+ m/z 542 (t = 1.28 min.).
Example 564
Figure imgf000223_0001
4-[2-(7-Bromo-2,3-dihydro-benzofuran-5-yl)-(S)-2-hydroxy-ethylamino]-3-{6-[(R)-2- fluoromethyl-moφholin-4-yl]-4-methyl- lH-benzimidazol-2-yl } - lH-pyridin-2-one and 4-[2-(7-bromo-2,3-dihydro-benzofuran-5-yl)-(S)-2-hydroxy-ethylamino]-3- { 6- [ S)-2-fluoromethyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - lH-pyridin-2- one: 1H NMR (400 MHz, CD3OD) δ 7.26 -7.40 (4H, m), 7.19 (IH, s), 6.24 (IH, d, 7 = 7.6 Hz), 4.81 - 4.82 (IH, m), 4.49 - 4.61 (4H, m), 3.18 - 4.18 (11H, m), 2.59 (3H, s). LCMS (M+H)+ m/z 598 (t = 1.32 min.).
Example 565
Figure imgf000223_0002
4-[2-(3-Chloro-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[('R)-2-hydroxymethyl- moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one and 4- [2-(3 - chloro -phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[('S -2-hydroxy-methyl-moφholin-4- yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.49 (IH, s), 7.23 - 7.37 (6H, m), 7.10 (IH, s), 6.24 (IH, d, 7= 7.6 Hz), 4.95 - 4.96 (IH, m), 4.19 (IH, m), 3.94 - 4.80 (2H, m), 3.59 - 3.71 (8H, m), 2.63 (3H, s). LCMS (M+H)+ m/z 510 (t = 1.21 min.).
Example 566
Figure imgf000224_0001
4-[2-(3-Bromo-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[(R)-2- hydroxymethyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one and 4-[2-(3-bromo-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3- { 6-[(S)-2- hydroxy-methyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.62 (IH, narrow d, 7= 2.0 Hz), 7.54 (IH, s), 7.29 - 7.36 (3H, m), 6.95 (IH, d, 7= 8.4 Hz), 6.23 (IH, d, 7= 7.6 Hz), 4.88 - 4.89 (IH, m), 3.56 - 4.19 (11H, m), 3.80 (3H, s), 2.60 (3H, s). LCMS (M+H)+ m/z 584 (t = 1.16 min.).
Example 567
Figure imgf000224_0002
4-[2-(3-Chloro-4-methoxy-ρhenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[ R)-2- hydroxymethyl-mθφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one and 4-[2-(3-chloro-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3- { 6-[(S)-2- hydroxy-methyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.68 (IH, s), 7.54 (IH, s), 7.45 (IH, s), 7.39 (IH, s), 7.30 - 7.32 (2H, m), 6.98 (IH, d, 7 = 8.4 Hz), 6.22 (IH, d, 7 = 7.6 Hz), 4.89 (IH, m), 4.11 - 4.19 (3H, m), 3.80 (3H, s), 3.49 - 3.72 (8H, m), 2.60 (3H, s). LCMS (M+H)+ m/z 540 (t = 1.09 min.).
Example 568
Figure imgf000225_0001
4-[2-(3-Chloro-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[(R)-2-methyl-moφholin-4- yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one and 4-[2-(3-chloro-phenyl)- (S)-2-hydroxy-ethylamino]-3-{6-[(S)-2-methyl-moφholin-4-yl]-4-methyl-lH- benzimidazol-2-yl}-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.75 (IH, s), 7.48 (IH, s), 7.42 (IH, s), 7.23 - 7.37 (4H, m), 6.24 (IH, d, 7 = 7.2 Hz), 4.94 - 4.97 (IH, m), 4.11 - 4.16 (3H, m), 3.61 - 3.68 (5H, m), 3.38 (IH, m), 2.64 (3H, s), 1.29 (3H, d, 7 = 6.4 Hz). LCMS (M+H)+ m/z 494 (t = 1.32 min.).
Example 569
Figure imgf000225_0002
4-[2-(3-Bromo-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[('R)-2-methyl- moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one and 4- [2-(3- bromo-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[('S)-2-methyl-moφholin- 4-yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.63 (IH, s), 7.62 (IH, s), 7.30 - 7.61 (3H, m), 6.95 (IH, d, 7= 8.4 Hz), 6.22 (IH, d, 7 = 7.6 Hz), 4.88 - 4.90 (IH, m), 4.08 - 4.18 (3H, m), 3.80 (3H, s), 3.61 - 3.67 (5H, m), 3.32 - 3.34 (IH, m), 2.60 (3H, s), 1.28 (3H, d, 7= 6.0 Hz). LCMS (M+H)+ m/z 568 (t = 1.31 min.).
Example 570
Figure imgf000226_0001
4-[2-(3-Chloro-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[('R)-2-methyl- moφholin-4-yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one and 4-[2-(3- chloro-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3- { 6-[(S)-2-methyl-moφholin- 4-yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one: *H NMR (400 MHz, CD3OD) δ 7.48 (IH, narrow d, 7 = 2.0 Hz), 7.40 (IH, br s), 7.31 (IH, narrow d, 7 = 2.0 Hz), 7.29 (IH, narrow d, 7 = 2.0 Hz), 6.97 (IH, d, 7 = 8.4 Hz), 6.22 (IH, d, 7 = 7.6 Hz), 4.87 - 4.90 (IH, m), 4.11 (IH, m), 3.95 - 4.01 (2H, m), 3.80 (3H, s), 3.47 (4H, m), 3.32 (IH, m), 3.25 (IH, m), 2.57 (3H, s), 1.27 (3H, d, 7= 6.4 Hz). LCMS (M+H)+ m/z 525 (t = 1.27 min.). Example 571
Figure imgf000227_0001
4-[2-(3-Chloro-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[(R)-2-methoxymethyl- moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one and 4- [2-(3 - chloro -phenyl)-(S)-2-hydroxy-ethyl amino] -3 - { 6- [(S)-2-methoxy-methyl-moφholin- 4-yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one: Η NMR (400 MHz, CD3OD) δ 7.49 (IH, narrow d, 7 = 2.0 Hz), 7.23 - 7.36 (6H, m), 6.22 (IH, d, 7 = 7.2 Hz), 4.93 - 4.96 (IH, m), 4.08 - 4.21 (1 IH, m), 3.38 (3H, s), 2.60 (3H, s), 1.27 (3H, d, 7 = 6.4 Hz). LCMS (M+H)+ m/z 524 (t = 1.35 min.).
Example 572
Figure imgf000227_0002
4-[2-(3-Bromo-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3- { 6-[(R -2- methoxymethyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one and 4-[2-(3-bromo-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[('S)-2- methoxymethyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.63 (IH, narrow d, 7= 2.0 Hz), 7.40 (IH, br s), 7.30 -7.36 (3H, m), 7.19 (IH, br s), 6.95 (IH, d, 7 = 8.4 Hz), 6.23 (IH, d, 7= 7.2 Hz), 4.87 - 4.89 (1H, m), 4.18 (IH, m), 3.97 - 4.03 (3H, m), 3.80 (3H, s), 3.54 - 3.66 (7H, m), 3.39 (3H, s), 2.59 (3H, s). LCMS (M+H)+ m/z 598 (t = 1.31 min.).
Example 573
Figure imgf000228_0001
4-[2-(3-Chloro-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[(R)-2- methoxymethyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one and 4-[2-(3-chloro-4-methoxy-phenyl)-(S)-2-hydroxy-ethylamino]-3-{6-[(S)-2- methoxymethyl-moφholin-4-yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.52 (IH, s), 7.46 (IH, narrow d, 7 = 1.6 Hz), 7.29 - 7.34 (3H, m), 6.99 (IH, d, 7= 8.4 Hz), 6.24 (IH, d, 7= 7.2 Hz), 4.89 (IH, m), 4.06 - 4.17 (3H, m), 3.81 (3H, s), 3.54 - 3.68 (8H, m), 3.38 (3H, s), 2.60 (3H, s). LCMS (M+H)+ m/z 554 (t = 1.28 min.).
Example 574
Figure imgf000228_0002
4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-[4-methyl-6-(4-methyl- piperazin-l-yl)-lH-benzoimidazol-2-yl]-lH-pyridine-2-one. 1H NMR (400 MHz, CD3OD) δ 7.46 (IH, s), 7.24-7.36 (m, 4H), 7.04 (2H, s), 6.22 (IH, d, 7= 7.64 Hz), 4.89 (IH, m), 3.30-3.82 (10H, m) 2.97 (3H, s), 2.57 (3H, -s). LCMS (M+H)+ m/z 493 (t = 1.56 min.)
Example 575
Figure imgf000229_0001
4-[2-(3-Bromo-4-methoxy-phenyl)-2(S)-hydroxy-ethylamino]-3-[4-methyl-6-(4- methyl-piperazin-l-yl)-lH-benzoimidazol-2-yl]-lH-pyridine-2-one. "H NMR (400 MHz, CD3OD) δ 7.61 (IH, d, 7 = 2.0 Hz), 7.32-7.37 (2H, m), 7.05 (2H, s), 6.97 (IH, d, 7= 8.52 Hz), 6.24 (IH, d, 7 = 7.64 Hz), 4.82 (lH,m), 3.82 (3H, s) 3.30-3.64 (10H, m), 2.98 (3H, s), 2.56 (3H, s) LCMS (M+H)+ m/z 567 (t = 1.53 min.)
The following examples (576-581) were prepared according to Scheme NE and EI and illustrate the acylation of a 4-amino-piperidine derivative
Example 576 (General procedure for Examples 576-581)
Figure imgf000229_0002
4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(acetamido)-piperidin-l-yl]- 4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one: A solution of the 4-amino- piperidine compound (~ 50-100 umol) in 5 ml MeOH is cooled to 0°C. Then -10 equivalent of Huenigs base and ~ 3 equivalent of acyl chloride are added. The vial is shaken once and allowed to stand at ambient temperature for 1 hour. Evaporation of volatiles and purification by prep. HPLC gives the 4-acyl-aminopiperidine compounds. 'H NMR (500 MHz, CD3OD) δ 8.06 (IH, s), 7.68 (IH, s), 7.49 (IH, s), 7.44 (IH, d, J = 7.6), 7.27 - 7.38 (4H, m), 6.33 (IH, d, 7 = 7.6 Hz), 4.96 (IH, dd, 7 = 4.1, 7.6 Hz), 4.17 (IH, m), 3.87 (4H, m), 3.65 (IH, dd, 7 = 4.2, 14 Hz), 3.58 (IH, dd, 7 = 7.8, 13.8 Hz), 2.72 (3H, s), 2.31 (2H, m), 2.19 (2H, m), 2.02 (3H, s). LCMS (M+H)+ m/z 535 (t = 1.03 min, YMC Xterra C18 S7 3.0 x 50 mm; 0 - 100% gradient over 1.5 min; 5 rnLΔnin flow rate).
Example 577
Figure imgf000230_0001
4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-hydroxyacetamido)- piperidin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one : LCMS (M+H)+ m/z 551 (t = 1.27 min.). HPLC t = 5.01 min, Waters Xterra C18 S5 4.6 x 30 mm; 0 - 100% gradient over 12 min; 5 mL/min flow rate.
Example 578
Figure imgf000230_0002
4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-fluoroacetamido)- piperidin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one : LCMS (M+H)+ m/z 597 (t = 1.31 min.). HPLC t = 6.90 min, YMC-Pack ODS-A 3.0 x 50 mm, 0 - 100% gradient over 12 min; 2.5 mL/min flow rate. Example 579
Figure imgf000231_0001
4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(acetamido)- piperidin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - lH-pyridin-2-one: LCMS (M+H)+ m/z 609 (t = 1.27 min.). HPLC t = 5.00 min, Waters Xterra C18 S5 4.6 x 30 mm; 0 - 100% gradient over 12 min; 5 mL/min flow rate.
Example 580
Figure imgf000231_0002
4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-hydroxyacetamido)- piperidin-1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - lH-pyridin-2-one: LCMS (M+H)+ m/z 595 (t = 1.30 min.). HPLC t = 5.09 min, Waters Xterra C18 S5 4.6 x 30 mm; 0 - 100% gradient over 12 min; 5 mL/min flow rate.
Example 581
Figure imgf000231_0003
4- [2-(3 -Chloro-phenyl)-2-hydroxy-ethylamino] -3 - { 6- [4-(2-fluoroacetamido)- piperidin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one: LCMS (M+H)+ m/z 553 (t = 1.35 min.). HPLC t = 5.90 min, YMC-Pack ODS-A 3.0 x 50 mm; 0 - 100% gradient over 10 min; 2.5 mL/min flow rate.
The following examples (582-584) were prepared according to Scheme NE and EI and illustrate the carbamoylation of a 4-amino-piperidine derivative
General Procedure for Examples 582-584 A solution of the 4-amino-piperidine compound (~ 50-100 umol) in 5 ml MeOH is cooled to 0°C. Then -10 equivalent of Huenigs base and ~ 3 equivalent of carbamoyl chloride are added. The vial is shaken once and allowed to stand at ambient temperature overnight. Evaporation of volatiles and purification by prep. HPLC gives the 4-carbamoyl-aminopiperidine compounds.
Example 582
Figure imgf000232_0001
4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2- methoxyethoxycarbamoyl)- piperidin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H- pyridin-2-one: ]H NMR (500 MHz, CD3OD) δ 7.90 (IH, s), 7.26 -7.52 (6H, m), 6.29 (IH, d, 7= 7.5 Hz), 4.97 (IH, dd, 7 = 4.1, 7.0 Hz), 4.20 (2H, m), 3.93 (IH, m), 3.60 - 3.84 (8H, m), 3.37 (3H, s), 2.69 (3H, s), 2.33 (2H, m), 2.13 (2H, m). LCMS (M+H)+ m/z 595 (t = 1.09 min, YMC Xterra C18 S7 3.0 x 50 mm; 0 - 100% gradient over 1.5 min; 5 mL/min flow rate). Example 583
Figure imgf000233_0001
4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(methoxycarbamoyl)- piperidm-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: Η NMR (500 MHz, CD3OD) δ 8.00 (IH, s), 7.63 (IH, s), 7.26 -7.49 (5H, m), 6.32 (IH, d, 7 = 7.6 Hz), 4.96 (IH, dd, 7= 4.2, 7.6 Hz), 3.93 (IH, m), 3.79 - 3.88 (4H, m), 3.67 (3H, s), 3.56 -3.65 (2H, m), 2.71 (3H, s), 2.32 (2H, m), 2.17 (2H, m). LCMS (M+H)+ m/z 551 (t = 1.07 min, YMC Xterra C18 S7 3.0 x 50 mm; 0 - 100% gradient over 1.5 min; 5 mL min flow rate).
Example 584
Figure imgf000233_0002
4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-fluoroethoxy carbamoyl)- piperidin-l-yl]-4-methyl-lH-benzoimidazol-2-yl}-lH-pyridin-2-one: "H NMR (500 MHz, CD3OD) δ 7.92 (IH, s), 7.26 -7.54 (6H, m), 6.3 (IH, d, 7= 7.5 Hz), 4.97 (IH, dd, 7 = 4.5, 7.5 Hz), 4.63 (IH, broad s), 4.53 (IH, d, J = 2.5 Hz), 4.32 (IH, broad s), 4.27 (IH, broad s), 3.93 (IH, m), 3.79 - 3.85 (4H, m), 3.58 - 3.72 (2H, m), 2.70 (3H, s), 2.33 (2H, m), 2.15 (2H, m). LCMS (M+H)+ m/z 583 (t = 1.29 min, YMC Xterra C18 S7 3.0 x 50 mm; 0 - 100% gradient over 2 min; 5 mL/min flow rate).
The following examples (585-590) were prepared according to Scheme NE and EI and illustrate the use of an alcohol as the nucleophile in Scheme NE Example 585 (General procedure for Examples 585-590)
Figure imgf000234_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(2-moφholin-4-yl- ethoxy)-lH-benzimidazol-2-yl]-lH-ρyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.53 (IH, s), 7.37 -7.39 (IH, m), 7.23 - 7.30 (3H, m), 7.01 (IH, s), 6.75 (IH, s), 6.21 (IH, d, 7= 7.2 Hz), 4.98 (IH, t, 7= 5.6 Hz), 4.40 (2H, br s), 3.97 (4H, br s), 3.45 - 3.73 (8H, m), 2.54 (3H, s). LCMS (M+H)+ m/z 524 (t = 1.24 min.).
Example 586
Figure imgf000234_0002
(S)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(2- moφholin-4-yl-ethoxy)-lH-benzimidazol-2-yl]-lH-pyridin-2-one : 1H NMR (400 MHz, CD3OD) δ 7.67 (IH, narrow d, 7 = 1.6 Hz), 7.36 (IH, dd, 7 = 1.6, 8.4 Hz), 7.25 (IH, d, 7= 7.2 Hz), 6.98 (IH, s), 6.92 (IH, d, 7= 8.4 Hz), 6.75 (IH, s), 6.22 (IH, d, 7 = 7.2 Hz), 4.89 - 4.92 (IH, m), 4.41 (2H, br s), 3.97 (4H, br s), 3.79 (3H, s), 3.34 - 3.66 (8H, m), 2.51 (3H, s). LCMS (M+H)+ m/z 598 (t = 1.22 min.). Example 587
Figure imgf000235_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(2-methoxy- ethoxy)-lH-benzimidazol-2-yl]-lH-pyridin-2-one : 1H NMR (400 MHz, CD3OD) δ 7.48 (IH, s), 7.25 -7.36 (4H, m), 6.96 (IH, s), 6.81 (IH, s), 6.20 (IH, d, 7 = 7.4 Hz), 4.90 - 4.93 (IH, m), 4.13 - 4.14 (2H, m), 3.76 - 3.77 (2H, m), 3.50 - 3.61 (2H, m), 3.43 (3H, s), 2.53 (3H, s). LCMS (M+H)+ m/z 469 (t = 1.52 min.).
Example 588
Figure imgf000235_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(2-hydroxy- ethoxy)-lH-benzimidazol-2-yl]-lH-pyridin-2-one : Η NMR (400 MHz, CD3OD) δ 7.43 - 7.45 (2H, m), 7.27 -7.34 (3H, m), 7.08 (IH, s), 7.04 (IH, narrow d, 7 = 1.0 Hz), 6.28 (IH, d, 7 = 7.6 Hz), 4.87 (IH, m), 4.13 (2H, t, 7 = 4.6 Hz), 3.92 (2H, t, 7 = 4.6 Hz), 3.45 - 3.54 (2H, m), 2.60 (3H, s). LCMS (M+H)+ m/z 455 (t = 1.35 min.).
Example 589
Figure imgf000236_0001
(S)-4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylaminoχ-3-[4-methyl-6-(3- moφholin-4-yl-propoxy)-lH-benzimidazol-2-yl]-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.66 (IH, s), 7.35 (IH, dd, 7 = 1.6, 7.9 Hz), 7.25 (IH, br s), 6.93 (IH, s), 6.91 (IH, s), 6.68 (IH, s), 6.19 (IH, br s), 4.86 (IH, m), 4.05 - 4.10 (4H, br s), 3.79 (3H, s), 3.17 - 3.73 (12H, m), 2.50 (3H, s). LCMS (M+H)+ m/z 612 (t = 1.16 min.).
Example 590
Figure imgf000236_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(3-moφholin-4-yl- propoxy)-lH-benzimidazol-2-yl]-lH-pyridin-2-one: 1H NMR (400 MHz, CD3OD) δ 7.52 (IH, s), 7.36 - 7.38 (IH, m), 7.23 - 7.30 (3H, m), 6.92 (IH, s), 6.67 (IH, s), 6.18 (IH, d, 7 = 6.9 Hz), 4.96 (IH, t, 7 = 5.9 Hz), 4.04 - 4.08 (4H, m), 3.82 (2H, m), 3.56 - 3.65 (8H, m), 3.15 - 3.18 (2H, m), 2.52 (3H, s). LCMS (M+H)+ m z 538 (t = 1.19 min.). The following examples (591-593) were prepared according to Scheme NE and EI wherein a cyano group (Scheme IN) is converted to an aldehyde which undergo reductive amination with an amine
Example 591 (General Procedure for Examples 591-593)
Figure imgf000237_0001
(S)-3-(4-Bromo-6-moφholin-4-ylmethyl-lH-benzimidazol-2-yl)-4-[2-(3-chloro- phenyl)-2-hydroxy-ethylamino]-lH-pyridin-2-one: To a solution of (S)-7-Bromo-2- {4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin-3-yl}-3H- benzimidazole-5-carbaldehyde (130 mg, 0.27 mmol) in methanol (60 mL) was added moφholine (0.2 mL, excess). The reaction mixture was stirred 1 h at room temperature. Then ΝaCΝBH3 (IM THF solution, 1.35 mL, 1.35 mmol) was added. The reaction mixture was stirred at room temperature overnight and concentrated in vacuo. The residue was purified by prep. HPLC to yield the title compound (68 mg, 45%).1H NMR (400 MHz, CD3OD) δ 7.83 (IH, s), 7.69 (IH, s), 7.68 (IH, s), 7.24 - 7.54 (4H, m), 6.30 (IH, d, 7 = 7.6 Hz), 4.89 - 5.02 (IH, m), 4.50 (2H, s), 3.66 - 3.71 (4H, m), 3.17 - 3.43 (6H, m). LCMS (M+H)+ m/z 558 (t = 1.41 min.).
Example 592
Figure imgf000237_0002
(S)-3-[4-Bromo-6-(4-methyl-piperazin- 1 -ylmethyl)- lH-benzimidazol-2-yl]-4-[2-(3- chloro-phenyl)-2-hydroxy-ethylamino]-lH-pyridin-2-one: 'H NMR (400 MHz, CD3OD) δ 7.70 (IH, s), 7.55 (2H, s), 7.42 (IH, d, 7 = 7.6 Hz), 7.24 - 7.32 (3H, m), 6.22 (IH, d, 7= 7.6 Hz), 5.01 (IH, t, 7= 6.2 Hz), 4.35 (2H, br s), 3.48 - 3.71 (10H, m), 2.98 (3H, s). LCMS (M+H)+ m/z 571 (t = 1.37 min.).
Example 593
Figure imgf000238_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(4-methyl- piperazin-l-ylmethyl)-lH-benzimidazol-2-yl]-lH-pyridin-2-one: Through a mixture of (S)-3-[4-Bromo-6-(4-methyl-piperazin-l-ylmethyl)-lH-benzimidazol-2-yl]-4-[2- (3-chloro-phenyl)-2-hydroxy-ethylamino]-lH-pyridin-2-one (80 mg, 0.14 mmol), tetramethyl tin (2.5 mL, excess), PdC12 (PPh3)4 (10 mg, 0.014 mmol), and KF (40 mg, 0.7 mmol) in DMF (2 mL) in a vial was bubbled nitrogen, sealed, and heated to 100°C for two days. The reaction mixture was passed through a small pad of celite. After concentration, the residue was purified by prep. HPLC to give the titled compound (34 mg, 48%). 1H NMR (400 MHz, CD3OD) δ 7.53 (IH, s), 7.45 (IH, s), 7.24 - 7.40 (4H, m), 7.09 (IH, s), 6.22 (IH, d, 7 = 7.6 Hz), 4.99 (IH, t, 7= 6.4 Hz), 4.04 (2H, br s), 3.61 - 3.74 (2H, m), 2.98 - 3.34 (8H, m), 2.80 (3H, s), 2.58 (3H, s). LCMS (M+H)+ m/z 507 (t = 1.29 min.).
The following examples (594-595) were prepared according to Scheme NE and EI and illustrate the use of tetrahydropyrimidine as the nucleophile in Scheme NE
Example 594
Figure imgf000239_0001
4-[2-(3-Chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-[4-methyl-6-(l,4,5,6- tetrahydropyrimidine- 1 -yl)- 1 H-benzoimidazol-2-yl] - lH-pyridine-2-one: * H NMR (400 MHz, CD3OD) δ 8.38 (IH, s), 7.52 (lH,s), 7.21-7.41 (5H, m), 7.04 (IH, d, 7 = 1.2Hz), 6.21(1H, d, 7 = 7.6Hz), 4.97 (IH, t, 7 = 4.9 Hz) 3.97(2H, m) 3.60-3.73 (2H, m),3.52(2H, t, J = 5.74 Hz) 2.60 (3H, s), 2.25 (2H, m) LCMS (M+H)+ m/z 477 (t = 1.79 min.)
Example 595
Figure imgf000239_0002
4-[2-(4-Methoxy-3-chloro-phenyl)-2(S)-hydroxy-ethylamino]-3-[4-methyl-6-(l,4,5,6- tetrahydropyrimidine- 1 -yl)- 1 H-benzoimidazol-2-yl] - 1 H-pyridine-2-one: ' H NMR (400 MHz, CD3OD) δ 8.37 (IH, s), 7.52 (lH,s), 7.04-7.52(4H, m), 7.04 (IH, d, 7 = 1.2Hz), 6.21(1H, d, 7 = 7.6Hz), 4.98 (IH, t, 7 = 4.92 Hz) 3.97(5H, m) 3.60-3.73 (2H, m),3.52(2H, t, J = 5.74 Hz), 2.60 (3H, s), 2.26 (2H, m). LCMS (M+H)+ m/z 507 (t = 1.67 min.)
Example 596 (Scheme NET) 4-[2-(3-Chloro-4-methoxy-phenyl)-2-hydroxy- ethylamino]-3-(4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5-dihydro- pyrrol-2-one
Figure imgf000240_0001
4-[2-(3-Chloro-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(l-tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-l- trifluoromethanesulfonyl-l,5-dihydro-pyrrol-2-one: 4-Hydroxy-3-(l-tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 ,5-dihydro- pyrrol-2-one (0.032 g, 0.059 mmol) was reacted as described in the procedure used to synthesize (S)-4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-3-(l-tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 - trifluoromethanesulfonyl-l,5-dihydro-pyrrol-2-one and by using acetonitiile instead of N,N-dimethylformamide. This gave the title material (0.018 g, 35%). LCMS (+ ESI, M+H+) m/z 730 ; 1H NMR (400 MHz, methanol-d4) δ (ppm): 1.55 (9H, s), 2.57 (3H, s), 3.20 (4H, m), 3.40 (2H, d, J=5.9 Hz), 3.85 (7H, m), 4.31 (IH, d, J=17.5 Hz), 4.44 (IH, d, J=17.6 Hz), 4.72 (IH, t, J=5.7 Hz), 7.02-7.04 (2H, m), 7.18 (IH, s), 7.30 (IH, d, J=8.4 Hz), 7.44 (IH, s).
Figure imgf000240_0002
4-[2-(3-Chloro-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin- 4-yl- 1 H-benzoimidazol-2-yl)- 1 ,5-dihydro-pyrrol-2-one: 4-[2-(3-Chloro-4-methoxy- phenyl)-2-hydroxy-ethylamino] -3-( 1 -tert-butyloxycarbonyl-4-methyl-6-moφholin-4- yl- 1 H-benzoimidazol-2-yl)- 1 -trifluoromethanesulfonyl- 1 ,5-dihydro-pyrrol-2-one (0.017 g, 0.023 mmol) was reacted according to the procedure used to synthesize (S)- 4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin-4-yl-lH- benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one and afforded the title material (0.0069 g, 59%) as a white solid. HPLC: 91% (220 nm), LCMS (+ESI, M+H+) m/z 498, HRMS calcd for C25H28N5O4Cl = 497.1830, found: 497.1815; IR v (cm 1): 3386, 1636, 1558; 1H NMR (400 MHz, methanol-d4) δ (ppm): 2.48 (3H, s), 3.11 (4H, br dd), 3.49 (IH, dd, J=13.9 and 6.8 Hz), 3.57 (IH, dd, J=13.9 and 4.3 Hz), 3.85-3.87 (7H, m), 4.14 (2H, 2d JAB=18.1 HZ), 4.85-4.88 (IH, m), 6.75 (IH, s), 6.92 (IH, s), 7.02 (IH, d, J=8.4 Hz), 7.33 (IH, dd, J=8.4 and 1.8 Hz), 7.47 (IH, d, J=1.8 Hz).
Example 597 (Scheme VET) 4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy- ethylamino]-3-(4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5-dihydro- pyrrol-2-one:
Figure imgf000241_0001
4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(l-tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 - trifluoromethanesulfonyl- l,5-dihydro-pyrrol-2-one: 4-Hydroxy-3-(l-tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 ,5-dihydro- pyrrol-2-one (0.022 g, 0.053 mmol) was reacted as described in the procedure used to synthesize (S)-4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-3-(l-tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 - trifluoromethanesulfonyl- l,5-dihydro-pyrrol-2-one and by using acetonitiile instead of N,N-dimethylformamide. This gave the title material (0.018 g, 44%). LCMS (+ ESI, M+H+) m/z 774; 1H NMR (400 MHz, methanol-dL.) δ (ppm): 1.55 (9H, s), 2.58 (3H, s), 3.20 (4H, br dd), 3.40 (2H, d, J=5.6 Hz), 3.85-3.86 (7H, m), 4.32 (IH, d, J=17.2 Hz), 4.44 (IH, d, J=17.2 Hz), 4.72 (IH, t, J=5.7 Hz), 7.01 (IH, d, J=8.3 Hz), 7.04 (IH, br s), 7.18 (IH, d, J=1.5 Hz), 7.34 (IH, dd, J=8.3 and 1.8 Hz), 7.62 (IH, d, J=1.8 Hz).
Figure imgf000242_0001
4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin- 4-yl-lH-benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one: 4-[2-(3-Bromo-4-methoxy- phenyl)-2-hydroxy-ethylamino]-3-(l-tert-butyloxycarbonyl-4-methyl-6-moφholin-4- yl- 1 H-benzoimidazol-2-yl)- 1 -trifluoromethanesulfonyl- 1 ,5-dihydro-pyrrol-2-one (0.018 g, 0.023 mmol) was reacted according to the procedure used to synthesize (S)- 4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin-4-yl-lH- benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one and afforded the title material (0.0068 g, 54%) as a beige solid. HPLC: 98% (220 nm), LCMS (+ESI, M+H+) m/z 542, 544, HRMS calcd for
Figure imgf000242_0002
= 541.1325, found: 541.1331; 1H NMR (400 MHz, methanol-cL,) δ (ppm): 2.48 (3H, s), 3.11 (4H, br dd), 3.49 (IH, dd, J=13.8 and 6.7 Hz), 3.57 (IH, dd, J=13.8 and 4.3 Hz), 3.84-3.87 (7H, m), 4.13 (2H, 2d, JAB=18.0 Hz), 4.85-4.94 (IH, m), 6.75 (IH, s), 6.92 (IH, s), 6.99 (IH, d, J=8.6 Hz), 7.37 (IH, dd, J=8.6 and 2.0 Hz), 7.64 (IH, d, J=2.0 Hz). Example 598 (Scheme NET) (S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4- methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 ,5-dihydro-pyrrol-2-one
Figure imgf000243_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(l-tert-butyloxycarbonyl-4- methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 -trifluoromethanesulfonyl- 1,5- dihydro-pyrrol-2-one: To a stirred solution of 4-hydroxy-3-(l-tert-butyloxycarbonyl- 4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 ,5-dihydro-pyrrol-2-one (0.034 g, 0.082 mmol) in dichloromethane (4 mL) at 0°C was added 2,6-di-tert-butyl-4 amino-pyridine (0.050 g, 0.246 mmol) followed by trifluoromethanesulfonic anhydride (30 μL, 0.180 mmol). The mixture was stirred at 0°C for about 10 minutes then water was added and this was extracted with dichloromethane (3x). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was then treated with a solution of (S)-2-amino-l-[3- chlorophenyl]ethanol (0.040 g, 0.233 mmol) in N,N-dimethylformamide (2 mL) at RT. The reaction was purifed by Prep HPLC (acetonitrile/ammonium acetate/water) to give the title material (0.012 g, 21%) along with 4-trifluoromethanesulfonyloxy-3- (l-tert-butyloxycarbonyl-4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5- dihydro-pyrrol-2-one (0.017 g, 38%), LCMS (+ESI, M+H+) m/z 547. Title material: LCMS (+ESI, M+H+) m z 700; 1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.46 (9H, s), 2.51 (3H, s), 3.13 (4H, br dd), 3.76 (5H, m), 4.30 (IH, m), 4.44 (2H, br s), 4.70 (IH, br qa), 5.80 (IH, d, J=4.3 Hz), 7.02 (IH, s), 7.07 (IH, s), 7.31-7.38 (3H, m), 7.43 (IH, s), 8.10 (lH, br t).
Figure imgf000244_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin-4-yl- lH-benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one: To a stirred solution of (S)-4-[2- (3-chloro-phenyl)-2-hydroxy-ethylamino]-3-(l-tert-butyloxycarbonyl-4-methyl-6- moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 -trifluoromethanesulfonyl- 1 ,5-dihydro- pyrrol-2-one (0.014 g, 0.020 mmol) in N,N-dimethylformamide (2 mL) was added saturated sodium bicarbonate (10 drops) and this was heated at 80°C for 6 hours. The reaction was cooled down to RT and water and dichloromethane were added. The pH of the aqueous phase was adjusted to 7 with 0.5 N hydrochloric acid. The aqueous phase was extracted with dichloromethane (5x). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue (0.012 g) was dissolved in dichloromethane (1.5 mL) and treated with SCX resin (0.750 g). This mixture was stirred at RT for 2 hours, then the resin was washed with dichloromethane, followed by ammonia (2N in methanol, diluted with dichloromethane (3 parts)). The ammonia solution was evaporated to give a residue (0.013 g) which was purified by Prep HPLC (acetonitrile/ammonium acetate/water) and afforded the acetic acid salt of the title material (0.0054 g, 51%) as a beige solid. HPLC: 95% (220 nm), LCMS (+ESI, M+H+) m/z 468, HRMS calcd for C24H26N5O3Cl = 467.1724, found: 467.1715; IR v (cm"1): 3382, 2922, 1636, 1597. 1H NMR (400 MHz, methanol-c ) δ (ppm): 1.89 (3H, s), 2.50 (3H, s), 3.11 (4H, br dd), 3.49 (IH, dd, J=13.7 and 7.9 Hz), 3.60 (IH, dd, J=13.7 and 4.1 Hz), 3.86 (4H, br dd), 4.15 (2H, d, JAB = 18.1 Hz), 4.92 (IH, dd, J=7.0 and 4.3 Hz), 6.76 (IH, br s), 6.94 (IH, br s), 7.27-7.39 (3H, m), 7.50 (IH, s). Example 599 (Scheme VIE) (S,S and S,R)-4-[2-(3-Chloro-phenyl)-2-hydroxy- ethylamino]-5-methyl-3-(4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5- dihydro-pyrrol-2-one:
Figure imgf000245_0001
(S,S and S,R)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-5-methyl-3-(l-tert- butyloxycarbonyl-4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 - trifluoromethanesulfonyl- l,5-dihydro-pyrrol-2-one: (S and R)-4-Hydroxy-5-methyl-3- (l-tert-butyloxycarbonyl-4-methyl-6-moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5- dihydro-pyrrol-2-one (0.023 g, 0.0536 mmol) was reacted as described in the procedure used to synthesize (S)-4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-3-(l- tert-butyloxycarbonyl-4-methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 - trifluoromethanesulfonyl- l,5-dihydro-pyrrol-2-one and by using acetonitiile instead of N,N-dimethylformamide. This gave the title material (0.014 g, 37%). LCMS (+ ESI, M+H+) m/z 714; 1H NMR (400 MHz, acetone-d6) δ (ppm): 1.50 (9H, s), 1.57 (3H, s), 2.54 (3H, s), 3.21 (4H, m), 3.81 (4H, m), 4.67 (2H, m), 4.98 (IH, m), 5.15 (IH, m), 7.08-7.35 (6H, m).
Figure imgf000245_0002
(S,S and S,R)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-5-methyl-3-(4-methyl- 6-moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one: (S,S and S,R)-4- [2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-5-methyl-3-(l-tert-butyloxycarbonyl-4- methyl-6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 -trifluoromethanesulfonyl- 1,5- dihydro-pyrrol-2-one (0.017 g, 0.024 mmol) was reacted according to the procedure used to synthesize (S)-4-[2-(3-chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6- moφholin-4-yl-lH-benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one and afforded the title material (0.0071 g, 63%) as a white solid. HPLC: 100% (220 nm), LCMS (+ESI, M+H+) m/z 482, HRMS calcd for C25H28N5O3Cl = 481.1881, found: 481.1868; 1H NMR (400 MHz, methanol-cU) δ (ppm): 1.41 and 1.45 (3H, 2d, J=6.8 and 6.6 Hz), 2.48 and 2.52 (3H, 2s), 3.11 (4H, m), 3.60 (IH, m), 3.69-3.77 (IH, m), 3.86 (4H, m), 4.29 and 4.47 (IH, 2m), 4.83-4.93 (IH, m overlapped by HDO), 6.76 (IH, br d), 6.92 (IH, br s), 7.26-7.48 (4H, m).
Example 600
Figure imgf000246_0001
[ 1 -(2- { 4- [2-(3 -Chloro-phenyl)-2-hydroxy-ethyl amino] -2-oxo- 1 ,2-dihydro-pyridin-3 - yl }-7-methyl-3H-benzoimidazol-5-yl)-piperidin-4-yl]-carbamic acid tetrahydro-furan- 3-ylmethyl ester : To a solution of the Tetrahydro-3-furanmethanol (lmmol) in THF (lml) was added NaH (lOOmg, 2.5mmol). After 5 min, a solution of Isocyanate compound (lOOmg, 0.19mmol) in THF (lml) was added. The reaction mixture was stirred at room temperature for 30min and concentrated, then purified by prep. HPLC to give the title compound (20mg, 17%). The title compound was converted to mono HCl salt via a procedure described in example 520. LCMS (M+H)+ m/z 621 (t = 1.4 min, YMC Xterra C18 7u 3.0 x 50 mm; 0 - 100% gradient over 2 min; 5 mL min flow rate).
Example 601
Figure imgf000247_0001
[ 1 -(2- {4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro-pyridin-3- yl }-7-methyl-3H-benzoimidazol-5-yl)-piperidin-4-yl]-carbamic acid 2-methoxy- propyl ester : To a solution of the (S)-(+)-2-Methoxypropanol (lmmol) in THF (lml) was added NaH (lOOmg, 2.5mmol). After 5 min, a solution of Isocyanate compound (lOOmg, 0.19mmol) in THF (lml) was added. The reaction mixture was stirred at room temperature for 30min and concentrated, then purified by prep. HPLC to give the title compound (25mg, 23%). The title compound was converted to mono HCl salt via a procedure described in example 520. LCMS (M+H)+ m/z 609 (t = 1.36 min, YMC Xterra C18 7u 3.0 x 50 mm; 0 - 100% gradient over 2 min; 5 mL/min flow rate).
Example 602; (S)-4-(2-{4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-2-oxo-l ,2- dihydro-pyridin-3-yl}-7-methyl-3H-benzoimidazol-5-yl)-piperazine-2-ethanol
Figure imgf000247_0002
(S)-4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-hydroxy-ethyl)- piperazin-l-yl]-4-methyl-lH-benzimidazol-2-yl}-lH-pyridin-2-one: 1HNMR (500 MHz, CD3OD) δ 7.60 (IH, s), 7.22 - 7.49 (4H, m), 7.04 (IH, s), 7.15 (IH, s), 6.36 (IH, d, 7 = 6.8 Hz), 4.88 (IH, m), 3.30 - 3.97 (14H, m), 2.63 (3H, s). LCMS (M+H)+ m/z 567 (t = 1.57 min.).
Figure imgf000248_0001
(S)-4-(2-{4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro-pyridin- 3-yl}-7-methyl-3H-benzoimidazol-5-yl)-piperazine-2-ethanol: To a stirred solution of 4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-piperazin-l-yl-lH- benzoimidazol-2-yl)-lH-pyridin-2-one (~ 2 TFA salt, 80 mg, -0.1 mmol) in methanol (2 mL) at 0 °C was added N,N-diisopropylethylamine (170 μL) and 2-fluoroethyl chloroformate (37 mg). The cooling bath was removed and the solution was stirred at room temperature for 30 minutes, after which LC/MS analysis showed the reaction to be complete. The reaction mixture was then purified on reverse phase preparative HPLC using a methanol / water / 0.1% trifluoroacetic acid gradient. The fractions were evaporated to give the title compound as a trifluoroacetic acid salt, which was dissolved in methanol and applied to a Narian Mega Bond-Elute SCX cartridge. Elution with methanol followed by 2.0 M ΝH3 / MeOH gave the free base (46.2 mg). This material was suspended in MeOH and 1.00 N aqueous HCl (1 equiv.) was added. The resulting solution was filtered through a 45 μm filter and evaporated to give the mono HCl salt of the title compound (46 mg): 1H NMR (500 MHz, CD3OD) δ 7.50 (brs, IH), 7.45 - 7.20 (m, 6H), 6.26 (d, IH, J = 7.7 Hz), 4.98 - 4.91 (m, IH), 4.64 (dm, 2H, J = 47.9 Hz), 4.39 (dm, 2H, J = 29 Hz), 3.95 - 3.50 (m, 10H), 2.63 (s, 3H); . LCMS (M+H)+ m z 569,571. Example 603
Figure imgf000249_0001
(S)-2-[4-(2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl } -7-methyl-3H-benzoimidazol-5-yl)-piperazin-l-yl]-acetamide Bis hydrochloride : 1H NMR (400 MHz, CD3OD) δ 7.45-7.24 (m, 5H), 7.12 (br s, IH), 7.09 (br s, IH), 6.24 (d, J=7.6Hz, IH), 4.95-4.88 (m, IH), 4.07 (s, 2H), 3.95-3.30 (m, 10H), 2.59 (s, 3H); LCMS (M+H)+ m/z 536.
Example 604
Figure imgf000249_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6[4-(2-methyoxy-ethyl)- piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl lH-pyridin-2-one bis-hydrochloride:
To a stirred solution of 4-[2-(3-Chloro-pheny)-2-hydroxy-ethylamino]-3-(4-methyl-6- piperazin-l-yl-lH-benzomidazol-2-yl)-lH-pyridin-2-one (100 mg, 0.123 mmol) in 1,4-dioxane (4 ml), ethanol (800ul), and methanol (800ul) was added Hunigs base (214ul). 2-Bromoethyl methyl ether (138ul) was added and the reaction was flushed with nitrogen and heated to 80°C until the reaction was complete as judged by LCMS. The reaction was cooled to room temperature and the solvents were evaporated in vacuo. The resulting residue was purified on reverse phase prepartive HPLC using a methanol/water/0.1% trifluoroacetic acid gradient. The fractions were evaporated to give the title compound as a trifluoroacetic acid salt. The trifluoroacetic acid salt of the pure title compound was dissolved in methanol and applied to a Narian Mega Bond-Elute SCX cartridge. Elution with methanol followed by 2.0 M ΝH3/MeOH gave the free base (27.3mg). This material was suspended in MeOH and 1.00 N aqueous HCl (2 equiv.) was added. The resulting solution was filtered through a 45 μm filter and evaporated to give the bis HCl salt of the title compound (25.8mg): 1H NMR (400 MHz, CD3OD) δ 7.45-7.23 (m, 5H), 7.16 (brs, IH), 7.08 (brs, IH), 6.25 (d,lH, J=7.6 Hz), 4.95-4.87 (m, IH), 3.92-3.20 (m, 14H), 3.44 (s, 3H), 2.60 (s, 3H); LCMS (M+H)+ /z 537,539.
Example 605
Figure imgf000250_0001
(S)-4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-{6[4-(2-methyoxy-ethyl)- piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl- 1 H-pyridin-2-one bis hydrochloride :
1H NMR (400MHz) δ 7.58 (brs, IH), 7.43-7.33 (m, 3H), 7.25 (t, IH), 7.15 (brs, IH),
7.09 (brs, IH), 6.24 (d, J=7.6 Hz, IH), 4.92-4.84 (m, IH), 3.83-3.24 (m, 14H), 3.43 (s, 3H), 2.60 (brs, 3H); LCMS [M+H]+ 581,583.
Example 606
Figure imgf000250_0002
(S)-4-[2-(3-Cynao-phenyl)-2-hydroxy-ethylamino]-3-{6[4-(2-methyoxy-ethyl)- piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl 1 H-pyridin-2-one bis hydrochloride : Η NMR (500MHz) δ 7.79 (brs, IH), 7.75 (d, J=8 Hz, IH), 7.66 (d, J=8 Hz, IH), 7.54 (t, J=7.5Hz, IH), 7.46 (d, J=7.5 Hz, IH), 7.2 (brs, IH), 7.14 (brs, IH), 6.33 (d, J=7.5 Hz, IH), 4.96-4.88 (m, IH), 3.85-3.26 (m, 14H), 3.45 (s, 3H), 2.62 (brs, 3H); LCMS [M+H]+ 528.
Example 607
Figure imgf000251_0001
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-hydroxy-ethyl)- piperadin- 1 -yl] -4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one bis hydrochloride: ]H NMR (500 MHz, CD3OD) δ 7.62 (s, IH), 7.50-7.28 (m, 6H), 6.32 (d, J=8 Hz, IH), 4.99-4.88 (m, IH), 3.82-3.58 (m, 8H), 2.71 (s, 3H), 2.22-1.60 (m, 7H); LCMS [M+H]+ 522, 524.
Example 608
Figure imgf000251_0002
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{4-methyl-6-[4-(2- methylsulf anyl-ethyl)-piperazin- 1 -yl] - 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one bis hydrochloride : The product of Example 550 (60mg) was treated with a solution of borane-tetrahydrofuran complex (1.0M in THF; 2.0mL) and stirred for 5 min. A solution of 0.1% TFA in 95:5 methanol/water was added (3.0mL) followed by 0.5mL neat TFA . The reaction mixture was then purified on reverse phase preparative HPLC using a methanol/water/0.1% trifluoroacetic acid gradient. The fractions were evaporated to give the title compound as a trifluoroacetic acid salt. The trifluoroacetic acid salt of the pure title compound was dissolved in methanol and applied to a Narian Mega Bond-Elute SCX cartridge. Elution with methanol followed by 2.0 M
ΝH3/MeOH gave the free base. This material was suspended in MeOH and 1.00 N aqueous HCl (2 equiv.) was added. The resulting solution was filtered through a 45 μm filter and evaporated to give 141 mg of the bis HCl salt of the title compound : 1H NMR (500 MHz, CD3OD) δ 7.47-7.40 (m, 2H), 7.37-7.25 (m, 3H), 7.19 (br s, IH), 7.11 (br s. IH), 6.28 (d, J=7.5Hz, IH), 4.95-4.88 (m, IH), 3.95-3.20 (m, 12H), 3.00- 2.93 (m, 2H), 2.62 (s, 3H), 2.22 (s, 3H); LCMS (M+H)+ m/z 553, 555.
Example 609
Figure imgf000252_0001
(S)4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(3R-methyl- piperaζin-l-yl)-lH-benζoimidaζol-2-yl]-lH-pyridin-2-one bis hydrochloride : Using the method described for the preparation of Example 517, R-2-methyl piperazine was used to give the titled compound; LCMS (M+H)+ m/z 493, 495.
Example 610
Figure imgf000252_0002
(S)4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-methoxy-ethyl)-3(R)- methyl-piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one bis hydrochloride : was prepared starting with (S)4-[2-(3-Chloro-phenyl)-2-hydroxy- ethylamino] -3-[4-methyl-6-(3R-methyl-piperazin- 1 -yl)- 1 H-benzoimidazol-2-yl] - 1 H- pyridin-2-one and using the method employed for the synthesis of Example 604. *H NMR (400 MHz, CD3OD) δ 7.45-7.23 (m, 5H), 7.16 (br s, IH), 7.09 (br s, IH), 6.24 (d, J=7.6Hz, IH), 4.95-4.87 (m, IH), 3.92-3.20 (m, 13H), 3.43 (s, 3H), 2.60 (s, 3H), 1.51 (d, 3H); LCMS (M+H)+ m/z 551, 553.
It is understood that the examples described above in no way serve to limit the true scope of this invention, but rather are presented for illustrative puφoses. All references cited herein are incoφorated by reference in their entirety.

Claims

We claim:
1. A compound selected from the group consisting of: 4-[2-(3-Chloro-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin-
4-yl- 1 H-benzoimidazol-2-yl)- 1 ,5-dihydro-pyrrol-2-one;
4-[2-(3-Bromo-4-methoxy-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin-
4-yl-lH-benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one;
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-(4-methyl-6-moφholin-4-yl- lH-benzoimidazol-2-yl)-l,5-dihydro-pyrrol-2-one;
(S,S and S,R)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-5-methyl-3-(4-methyl-
6-moφholin-4-yl- 1 H-benzoimidazol-2-yl)- 1 ,5-dihydro-pyrrol-2-one;
[ 1 -(2- { 4- [2-(3 -Chloro-phenyl)-2-hydroxy-ethylamino] -2-oxo- 1 ,2-dihydro-pyridin-3- yl }-7-methyl-3H-benzoimidazol-5-yl)-piperidin-4-yl]-carbamic acid tetrahydro-furan- 3-ylmethyl ester;
[ 1 -(2- { 4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo- 1 ,2-dihydro-pyridin-3- yl }-7-methyl-3H-benzoimidazol-5-yl)-piperidin-4-yl]-carbamic acid 2-methoxy- propyl ester;
(S)-2-[4-(2-{4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-2-oxo-l,2-dihydro- pyridin-3-yl}-7-methyl-3H-benzoimidazol-5-yl)-piperazin-l-yl]-acetamide Bis hydrochloride;
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6[4-(2-methyoxy-ethyl)- piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl lH-pyridin-2-one bis-hydrochloride;
(S)-4-[2-(3-Bromo-phenyl)-2-hydroxy-ethylamino]-3-{6[4-(2-methyoxy-ethyl)- piperazin- 1 -yl]-4-methyl 1 H-benzoimidazol-2-yl- 1 H-pyridin-2-one bis hydrochloride;
(S)-4-[2-(3-Cynao-phenyl)-2-hydroxy-ethylamino]-3-{6[4-(2-methyoxy-ethyl)- piperazin- 1 -yl] -4-methyl- 1 H-benzoimidazol-2-yl 1 H-pyridin-2-one bis hydrochloride ;
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-hydroxy-ethyl)- piperadin- 1 -yl]-4-methyl- 1 H-benzimidazol-2-yl } - 1 H-pyridin-2-one bis hydrochloride;
(S)-4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{4-methyl-6-[4-(2- methylsulfanyl-ethyl)-piperazin- 1 -yl]- 1 H-benzoimidazol-2-yl } - 1 H-pyridin-2-one bis hydrochloride; (S)4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-[4-methyl-6-(3R-methyl- piperazin-l-yl)-lH-benzoimidazol-2-yl]-lH-pyridin-2-one bis hydrochloride; and (S)4-[2-(3-Chloro-phenyl)-2-hydroxy-ethylamino]-3-{6-[4-(2-methoxy-ethyl)-3(R)- methyl-piperazin- 1 -yl]-4-methyl- 1 H-benzoimidazol-2-yl } - lH-pyridin-2-one bis hydrochloride.
2. Use of a compound according to claim 1 in the manufacture of a composition for use in treating cancer.
3. Use of a compound according to claim 1 in the manufacture of a composition that is used in combination with other anticancer agents for the treatment of cancer.
4. A pharmaceutical composition comprising a compound according to claim 1 in a pharmaceutically acceptable carrier.
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CN117069696B (en) * 2023-08-17 2024-04-26 中国药科大学 Double-target small molecule inhibitor and preparation method and application thereof

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AU2003282891A8 (en) 2004-04-23
US7223757B2 (en) 2007-05-29
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