CA2250143A1 - Inhibitors of farnesyl-protein transferase - Google Patents

Abstract

The present invention is directed to compounds which inhibit farnesyl-protein transferase (FTase) and the farnesylation of the oncogene protein Ras. The invention is further directed to chemotherapeutic compositions containing the compounds of this invention and methods for inhibiting farnesyl-protein transferase and the farnesylation of the oncogene protein Ras.

Description

- TITLE OF THE INVENTION
INHIBITORS OF FARNESYL-PROTEIN TRANSFERASE

BACKGROUND OF THE ~VENTION
The present invention relates to compound.s which inhibit farnesyl protein transferase, a protein which is implicated in the oncogenic pathway mediated by Ra.s. The Ra.s protein,s (Ha-Ras, Ki4a-Ras, Ki4b-Ras and N-Ras) are part of a signalling pathway that links cell surface growth factor receptors to nuclear signals initiating cellular proliferation. Biological and biochemical .studie.~i of Ras action indicate that Ra.s functions like a G-regulatory protein. In the inactive ~state, Ras i.s bound to GDP. Upon growth factor receptor activation R~
is induced to exchange GDP for GTP and undergoes a conformational change. The GTP-bound form of Ras propagates the growth ,stimulatory signal until the ~-ignal is terminated by the intrinsic GTPase activity of Ras, which returns the protein to its inactive GDP
bound form (D.R. Lowy and D.M. Willumsen, AM~1. Rel!. BiOChenl.
~2:~51-~91 (1993)). Mutated ra~ genes (Ha-7a~, Ki4a-ru~, Ki4b-ras and N-ras) are found in many human cancers, including colorectal carcinoma, exocrine pancreatic carcinoma, and myeloid leukemia.s.
The protein products of the~e genes are defective in their GTPase activity and con.stitutively transmit a growth .stimulatory signal.
Ras must be localized to the plasma membrane for both normal and oncogenic functions. At least 3 post-translational modifications are involved with Ras membrane localization, and all 3 modifications occur at the C-te~ninu,s of Ras. The Ra~ C-terminus contain~ a sequence motif termed a "CAA~" or "Cys-Aaal-Aaa2-Xaa"
box (Cy.s i.s cy.steine, Aaa i~i an aliphatic amino acid, the Xaa i~ any amino acid) (Willumsen et al., Nature 310:5~3-5~6 ( 19R4)). Depend-ing on the .specific .sequence? this motif serves as ~ ~;ignal ~sequence for the enzymes farnesyl-protein tran,sferase or geranylgeranyl-protein transferase, which catalyze the alkylation of the cy,steine residue of the CAAX motif with a Cls or C2() isoprenoid, re.spectively. (S. Clarke.~
AMM. Rel~. B~ he~7?. fjl :355-3~6 (1992); W.R. Schafer ~nd J. Rine, Ann.

CA 022~0143 1998-09-2~

WO g7/36583 PCT/US97/05170 Rev. Genetics 30:209-237 (1992)). Ra.s proteins are known to undergo post-translational farnesylation. Other farnesylated proteins include the Ras-related GTP-binding proteins ~such as Rho, fungal mating factors, the nuclear lamins, and the gamma subunit of transducin. James, et al., 5 J. Bivl. Chem. 269, 14182 (1994) have identified a peroxilsome associated protein Pxf which i~ also ~arnesyl-ated. James. et al., have also .~uggested that there are farnesylated proteins of unknown structure and function in addition to those listed above.
Inhibition of farnesyl-protein transfera~e has been shown to block the growth of ~as-transformed cells in soft ag~r and to modify other aspects of their transformed phenotype. It has also been demonstrated that certain inhibitors of farne!iyl-protein transfera.se selectively block the processing of the Ras oncoprotein intracellularly 15 (N.E. Kohl et al., Science, 260:1934-1937 (1993) and G.L. Jame.s et al., Science, 260:1937-1942 (1993). Recently, it has been ~hown that an inhibitor of farnesyl-protein transferase blocks the growth of ras-dependent tumors in nude mice (N.E. Kohl et al., P~ oc. Natl. Acad. S~i U.S.A., 91:9141-9145 (1994) and induces regression of m~mm~ry and 20 salivary carcinomas in )~as transgenic mice (N.E. Kohl et al., Natu) e Medicine, 1 :792-797 (1995).
Indirect inhibition of farnesyl-protein transferase in vivo has been demonstrated with lovastatin (Merck & Co., Rahway, NJ) and compactin (Hancock et al., ihid; Casey et al., ihid; Schafer 25 et al., Science 245:379 (1989)). These drugs inhibit HMG-CoA
reductase, the rate limiting enzyme for the production of poly-isoprenoids including farnesyl pyrophosphate. Farnesyl-protein transfera.se utilize~s farnesyl pyrophosphate to covalently modify the Cys thiol group of the Ras CAAX box with a farne.syl group (Reiss 30 et al., Cell, 62~ (1990); Schaber et al., J. Biol. Chem.. 265:14701-14704 (1990); Schafer et al., Science, 249:1133-1139 (1990); Manne et al., P~oc. Natl. Acad. Sci USA, ~7:7541-7545 (1990)). Inhibition of farnesyl pyrophosphate biosynthesis by inhibitin~ HMG-CoA reductase blocks Ras membrane localization in cultured cells. However, direct CA 02250l43 l998-09-25 inhibition of farnesyl-protein transfera~se would be more ~specific, and thu,s preferable.
Inhibitors of farne~syl-protein transferase (FPTase) have been described in two general classe~s. The fir~st are analogs of farnesyl diphosphate (FPP), while the second cla,ss of inhibitors is related to the protein substrates (e.g., Ras) for the enzyme. The peptide derived inhibitor.s that have been de~scribed are generally cysteine containing molecule~s that are related to the CAAX motif that is the signal for protein prenylation. (Schaber et al., ibid; Reiss et. c~l., ihid; Reiss et al., PNAS, 88:732-736 (1991)). Such inhibitors may inhibit protein prenylation while serving as alternate substrates for the famesyl-protei tran~sfera~se enzyme, or may be purely competitive inhibitor~s (U.S.
Patent 5,141,~51, University of Texa~s; N.E. Kohl et al., Science, 260: 1934- 1937 (1993); Graham, et al., J. Med. Chem., 37, 725 (1994)).
It has recently been reported that FPT-ase inhibitors also inhibit the proliferation of vascular ,smooth muscle cells and are therefore useful in the prevention and treatment of arteriosclero~sis and diabetic disturbance of blood vessel,s (JP H7-112930).
It ha,s recently been disclosed that certain tricyclic compound,s which optionally incorporate a piperidine moiety are inhibitors of FPTase (WO 95/10514, WO 95/10515 and WO 95/10516). Imidazole-containing inhibitors of farne.syl protein transferase have al~so been disclo~sed (WO 95/09001 and EP 0 675 112 Al).

SUMMARY OF THE INVENTION
The present invention addresse~s a compound of formula 1:

- V Al (CR1 a2) A2(C R 1b2)n -(W)- (CR 2~A - (CR 2)p I

CA 022~0143 1998-09-2~

or a pharmaceutically acceptable salt thereof, wherein:

Rla, R1b and R2 are independently selected from the group consisting of: hydrogen, aryl, heterocyclyl, C3-Clo S cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(o)m-~(R~)2NC(O)-, R~C(O)NR8-, CN, NO2, (R~)2NC(NRg)-, R8C(O)-, R~OC(O)-, N3, -N(R~)2, R9OC(o)NR8- and Cl-C6 alkyl, unsubstituted or substituted by 1-3 groups selected from the group con.sisting of:
halo. aryl, heterocyclyl, C3-Clo cycloalkyl, C2-c6 alkenyl, C2-C6 10 alkynyl, R8O-, R9S(o)m-~ R~C(O)NR~-, CN, (R~)2NC(NR~)-, R~C(O)-, R8OC(O)-, N3, -N(R8)2 and R9OC(o)NR~-;

R3 and R4 are independently selected from the group consisting of: H, F, Cl, Br, -NR82, CF3, NO2, R8O-, R9S(o)m-~
15 (R~)2NC(O)-, R8C(O)NH-, H2NC(NH)-, R~C(O)-, R~sOC(O)-, N3, CN, R9OC(o)NR8-, Cl-C20 alkyl, substituted or unsubstituted aryl and substituted or unsubstituted heterocyclyl;

A3 is selected from: ~NRSs(o)m- or -S(O)mNRS-~
20 with m equal to 0, 1 or 2, and RS selected from the group consisting of:hydrogen, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, un~substituted or substituted C3-Clo cycloalkyl, ~md Cl-C6 alkyl, unsubstituted or substituted with 1-3 members selected from the group consisting of: unsubstituted or substituted aryl, unsubstituted or 25 sub.stituted heterocyclyl, unsubstituted or substituted C3-Clo cycloalkyl, -N(R~)2, -CF3, -NO2, (R~)O-, (R9)S(o)m-~ (R~)C(O)NH-, H2NC(NH)-, (R~)C(O)-, (R~)OC(O)-, N3, CN and (R9)OC(O)NR~-;

R6 and R7 are independently selected from the group 30 consisting of: hydrogen, ~ryl. heterocyclyl, C3-CIo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1 6 perfluoroalkyl, F, Cl, Br, R~O-, R9S(o)m-, R~sC(O)NRPi-, CN, N02, (RX)2NC(NR~)-, R~C(O)-, R~OC(O)-, N3, -N(R~s)2, R90C(o)NR~- and Cl-C6 alkyl unsubstituted or substituted by 1-3 groups selected from:

CA 022~0143 1998-09-2~

aryl, heterocyclyl, C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R~O-, R9S(o)m-~ RXC(O)NRx-, CN, (RP~)2NC(NR~)-, R~C(O)-, RXOC(O)-, N3, -N(RX)2 and R9OC(o)NR~¢-;
s each R~s ils independently selected from hydrogen, Cl-C6 alkyl, aryl and aralkyl;

each R9 is independently selected from Cl-C6 alkyl and 10 aryl;

A 1 and A2 are independently ,selected from the group con,sisting of: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR~-, -NR~C(O)-, -O-~ -N(R8)-, -S(O)2N(RX)-, -N(R~)s(o)2-~ and S(O)m;~5 X represents aryl or heteroaryl;

V is ,selected *om the group consisting of: hydrogen, heterocyclyl, aryl, Cl-C2o alkyl wherein from 0 to 4 carbon atoms are 20 replaced with a heteroatom selected from O, S, and N~ and C2-C20 alkenyl, provided that V i,s not hydrogen if Al is S(O)m and V is not hydrogen if ~1 is a bond, n is 0 and A2 i.s S(O)m;

W represents heterocyclyl;~5 each n and p independently represents 0, 1, 2, 3 or 4;
r is 0 to 5, provided that r is 0 when V is hydrogen, and ti,s0Or 1.

~0 DETAILED DESCRIPTION OF THE INVENTION
The compounds of thi,s invention are u~seful in the inhibition of farnesyl-protein tran.sfera.se and the farnesylation of the oncogene protein Ras, and thu~ are useful for the treatment of cancer.

CA 022~0143 1998-09-2~

The compounds of the present invention may have asymmetric centers and occur as racemates, racemic mixtures and as individual diastereomers, with all possible isomers, including optical isomers, being included in the present invention.
When any variable (e.g. aryl, heterocycle, Rl, R2 etc.) occurs more than one time in any constituent, each definition is independent.
The term "alkyl" and the alkyl portion of alkoxy, aralkyl and similar terms, is intended to include branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atom~i~ or 1-6 carbon atoms if unspecified. Cycloalkyl means 1-2 cabocyclic rings which are saturated and contain from 3-10 atoms.
"Halogen" or "halo" as used herein means fluoro, chloro, bromo and iodo.
As u~sed herein, "aryl" and the aryl portion of aralkyl, are intended to mean any ~stable monocyclic or bicyclic carbon ring of up to 7 members in each ring, wherein at least one ring i~s aromatic. Examples of such aryl elements include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl. A preferred aralkyl group is benzyl.
The terms heterocyclyl, heterocycle and heterocyclic, as used herein, mean a 5- to 7-membered monocyclic or ~- to I 1-membered bicyclic heterocyclic rings, either saturated or unsaturated, aromatic, partially aromatic or non-aromatic, and which consist of carbon atoms and from one to four heteroatom,s selected from the group consisting of N, O, and S. Thus, it includes any bicyclic group in which any of the above-defined heterocyclic ring~s i,s fused to a benzene ring.
The ring or ring system may be attached at any heteroatom or carbon atom which results in a stable structure, and may contain 1-3 carbonyl groups. Examples of ~such heterocycles include, but are not limited to, azepinyl, benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl~ benzothiopyranyl, benzofuryl, benzothiazolyl, benzothienyl~ benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, CA 022~0143 1998-09-2~

dihydrobenzothiopyranyl sulfone, furyl, imidazolidinyl, imidazolinyl, imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, isothiazolidinyl, morpholinyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 2-oxopiperazinyl, 2-oxopiperdinyl, 2-oxopyrrolidinyl, piperidyl, piperazinyl, pyridyl, pyrazinyl, pyrazolidinyl, pyrazolyl, pyridazinyl, pyrimidinyl, pyrrolidinyl, pyrrolyl, ~uinazolinyl, quinolinyl, quinoxalinyl, tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiazolyl, thiazolinyl, thienofuryl, thienothienyl, and thienyl.
"~Ieteroaryl" i~ a .sublset of heterocyclic, and means a monocyclic or bicyclic ring system, with up to 7 members in each ring, wherein at least one ring is aromatic and wherein from one to four carbon atoms are replaced by heteroatoms selected from the group consisting of N, O, and S. Example~i include benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, furyl, imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, iso~uinolinyl, isothiazolyl, naphthyridinyl, oxadiazolyl, pyridyl, pyrazinyl, pyrazolyl.
pyridazinyl, pyrimidinyl, pyrrolyl, quinazolinyl, ~uinolinyl, quinoxalinyl, tetrahydroiso4uinolinyl, tetrahydro4uinolinyl, thiazolyl, thienofuryl, thienothienyl and thienyl.
Lines drawn into ring systems from substituents indicate that the bond may be attached to any of the substitutable ring atoms.
The term "substituted" as used with respect to, e.g., substituted alkyl, substituted aryl, substituted heterocyclyl and substituted cycloalkyl mean alkyl, aryl, heterocyclyl and cycloalkyl group~, respectively. having from 1-3 substituents which are selected from: halo, aryl, heterocyclyl, C3 1~) cycloalkyl, Cl ~ alkyl, C2 ~, alkenyl, C2 ~, alkynyl, RXO-, R9S(o),n-, RXC(O)NRX-, CN, (RX)~NC(NRx)-, RXC(O)-, RXOC(O)-, N~" -N(RX)2 and R~OC(O)NRX-.

CA 022~0143 1998-09-2~

Preferably 1-2 groups are present on substituted alkyl, substituted aryl, substituted heterocyclyl and sub.stituted cycloalkyl, which are selected from: halo, aryl, RX0-, CN, R~C(0)- and -N(RX)2.
Preferably, R1a, Rlb and R2 are independently selected S from: hydrogen, -N(R~)2, R~C(O)NR8- or unsubstituted or substituted Cl-C6 alkyl wherein the substituent on the substituted Cl-C6 alkyl is selected from unsubstituted or substituted aryl, -N(R~)2, R80- and R~C(O)NR~- Preferably, R3 and R4 are selected from: hydrogen and Cl-C6 alkyl.
Preferably, A3 represents NRSS(O),n, wherein m represents 2 and R5 represents hydrogen.
Preferably, R6 represents hydrogen, un~sub~stituted or .substituted Cl-C6 alkyl.
Preferably R7 repre,sents H or un,substituted C~ ~ alkyl.
Preferably, R% represents H or C~ ~ alkyl, and R9 is C
alkyl.
Preferably, Al and A2 are independently selected from:
a bond, -C(O)NR8-, -NR~C(0)-, -0-, -N(RP~)-, -S(0)2N(R~)- and-N(R~S(0)2--Preferably X represents aryl and mo.st preferably phenyl.
Preferably. V is selected from hydrogen, heterocyclyl and aryl. More preferably V is phenyl.
Preferably, W is heterocyclyl selected from imidazolinyl, imidazolyl, oxazolyl, pyrazolyl, pyyrolidinyl, thiazolyl and pyridyl.
More preferably, W is selected from imidazolyl and pyridyl.
Preferably, m is 2.
Preferably n and p are independently 0, 1, 2 or 3.
Preferably t is 1.
A ,sub,set of compounds of the invention is repre.sented by formula Ia:
-~ (R6) R7 V - A1(CR1a2)nA2(CR1b2)~N~_~J R3 (CR 2)p--A - (CR 2)p{

whereln:
R3, R4, A3, RX, R9, m, n, p and r are as originally defined;

each Rla and R2 is independently ~elected from hydrogen and C I -C6 alkyl;

each Rlb i,~i independently selected from: hydrogen, aryl, 10 heterocyclyl, C3 10 cycloalkyl, C2 ~ alkenyl, R~O-, -N(R~)2 and Cl-C6 alkyl unsubstituted or.substituted by aryl, heterocyclyl, cycloalkyl, alkenyl, R8O- and -N(R8)2;

RS is selected from the ~roup consisting of: hydrogen and 15 Cl-C6 alkyl, unsubstituted or substituted with 1-3 member~i ~elected from the group con.sisting of: unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl. unsub~stituted Ol .~iubstituted C3-Clo cycloalkyl, -N(R~)2, -CF3, -N02, (R~)O-, (R9)S(o),~,-, (R8)C(O)NH-, H2NC(NH)-, (R~)C(O)-, (R~¢)OC(O)-, N3, CN and 20 (R9)OC(O)NR8-;

R6 i~s .~elected from: hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, ~, Cl, R~O-, R~C(O)NRP~-, CN, N02, (R~)2N-C(NR~)-, R~sC(O)-, R~OC(O)-, 2~ -N(R8)2, or R9OC(o)NRg-, and Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl, R~O-, R~C(O)NR~-, (R~)2N-C(NR~)-, R~SC(O)-, R~OC(O)-, -N(RX)2 and R9OC(o)NR~-;

CA 022F,0 143 1998 - 09 - 2F, WO 97136583 PCT/US97tO5170 - I O-R7 is hydrogen or unsubstituted Cl ~ alkyl;

A I and A2 are independently selected from: a bond, -CH=CH-, -C_C-, -C(O)-, -C(O)NR8-, O, -N(R~)- and S(O)m;

and V is selected from: hydrogen; aryl; heterocyclyl ,selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, i.soquinolinyl and thienyl; Cl-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a a hetero-10 atom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if AI i~i a bond and A2 i,'; S(O)m.
A ~econd subset of compound.s of the present invention is repre,sented by formula Ib:

V - A1 (CRl a2)nA2(CR 1b2)n ~W)~ (C R22)F NRs-s(o)m- (CR22h~CI~) Ib R4 wherein:

R1a, Rlb, R2, Al, A2, R3, R4, R5, R6, R~, R~, m, n, p and r 20 are as originally defined;

R7 is selected from: hydrogen and unsubstituted Cl-C6 alkyl;

2~ V is selected from: hydrogen, heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl. indolyl, quinolinyl, isoquinolinyl, and thienyl, aryl, Cl-C20 alkyl wherein from 0 to 4 carbon atom~i are replaced with a heteroatom selected from O, S, and N, and C2-C~o alkenyl, provided that V is not hydrogen if A~ S(O)m and V i~ not hydrogen if Al is a bond, n i~ 0 and A2 is S(O)m; and S ~I reprelsents heterocyclyl ~elected from pyrrolidinyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl and i~oquinolinyl.

A third embodiment of the invention i~s de~cribed in 10 accordance with formula Ic:
H

)~N

(C R 2)p -A - (C R 2)p~

whereln:

each R2 i~j independently selected from hydrogen and Cl-C6 alkyl;

R3, R4, A3, RX, R9, m and p are as originally defined;

each R5 i,s selected from: hydrogen and Cl-C6 alkyl unsubstituted or substituted with 1-3 group~ selected from un_stituted or sub~tituted aryl, unsubstituted or substituted heterocyclyl, unsubstituted or ~ubstituted C3-clo cycloalkyl, -N(R~)2~ -CF3, -NO2.
(R~)O-, (R9)S(o)m-, (R~)C(O)NH-, H2NC(NH)-, (R~)C(O)-, 2~; (R~)OC(O)-, N~,~ -CN ~nd (R9)OC(O)NR~-;

and R6 i~ ~;elected from the group con~ ting of: hydrogen~
Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl, R8O-, R~C(O)NR8-, CN, NO2, (R~)2N-C(NRX)-~ R~C(O)-, R80C(O)-, -N(RP~)2, or R9OC(o)NR8- and CI-C6 alkyl substituted by Cl-C6 perfluoroalkyl, R8O-, R~C(O)NR8-, (R8)2N-C(NR8)-, R~C(O), R8OC(O)-, -N(R~)2 or R9OC(o)NR8-.

A fourth subset of compounds of the invention is represented by formula Id:
H

(CR22)p 7~3 - (CR22)p{
n4 NC n wherein:
each R2 is independently selected from: hydrogen and Cl-C6 alkyl;

R3 and R4 are independently selected from H, F, Cl, Br, N(RX)2, CF3, NO2, (R8)o-, (R9)S(o)m-~ (R8)C(O)NH-, H2N-C(NH)-, 1~ (R8)C(O)-, (R~)OC(O)-, N3, CN, (R9)OC(O)NR~-, Cl-C20 alkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclyl;

A3 represents -NR5-S(o),- or-S(O)m-NR5-;
R5 is selected from: hydrogen and Cl-C6 alkyl, unsub~tituted or substituted witha group selected from unsubstituted Ol substituted aryl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted C3-Clo cycloalkyl, N(R~)2, CF3, NO2, (R~)O-, 2~ (R9)S(O)m-, (R~)C(O)NH-, H2N-C(NH)-, (R~)C(O)-, (R~)OC(O)-, N3, CN (R9)OC(O)NR~¢-;

and R~, R9, m and p are a!i originally defined.

Specific examples of compound.s of the invention are:

G--~N ~ N H SO2~

N C I
NC~ ~SO2NH~ "(~

N

NC ~ ~ NHSO

N

NC ~ ~ SO2NH

N

Nl ~ ~~ ~CI ~N o~S--N~ I

NC NC

\~\N--'S~

NC Cl NC ~N N(CH3)SO~

N~ Cl NC ~3\ ~SO2N(CH3 N

NC ~3~ N(CH3)S~

;~ N~

NC ~'N ~SO2N(CH3)~

N

CA 022~0143 1998-09-2~

WO 97/36~83 PCT/US97/05170 ~ O~ ~CI ~ of S-N~

NC NC

~N-S~

NC

and the pharmaceutically acceptable salt,s thereof.
S The pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds of this invention a~s forrned, e.g., from non-toxic inorganic or organic acid~s. For example, such conventional non-toxic salts include those derived from inorganic acids such a~i hydrochloric, hydrobromic, ~sulfuric, sulfamic, phosphoric, nitric and the like: ~nd the salt~i prepared from organic acid.s such as acetic, propionic, ~uccinic, glycolic, .stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, ,salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.
The pharmaceutically acceptable .salt.s of the compound.s of thi.s invention can be synthe~;ized from the compound.s of thi.s invention which contain a basic moiety by conventional chemical - methods. Generally, the .salts are prepared either by ion exchange chromatography or by reacting the free ba~e with stoichiometric amount.s or with ~n exce~,s of the desired .salt-forming inorganic or organic acid in a suitable .solvent or variou.s combination~s of ~solvent~s.

CA 022~0143 1998-09-2~

Reaction.s u.sed to generate the compounds of this invention are prepared by employing reactions a,s shown in Schemes 1-7, in addition to other standard manipulations such as ester hydrolysis, cleavage of protecting groups, etc., as may be known in the literature 5 or exemplified in the experimental procedures. Substituents R and R CH2-, as shown in the Scheme,s, repre,sent the sub,stituents R~, R9 and other,s, depending on the compound of the instant invention that i,s being ~synthesized. The variable p' repre,sents p-l.
These reaction,s may be employed in a linear ,sec~uence to 10 provide the compounds of the invention or they may be used to synthesize fragments which are ,subsequently joined by the alkylation reactions described in the Schemes.

Synopsi,s of Scheme.s The re~luisite intermediate.s are commercially available or can be prepared according to literature procedures. The Schemes illustrate the synthesi,s of certain preferred embodiments of the instant invention, wherein the variable W is present a,s an imidazolyl moiety that is sub,stituted. Substituted protected imidazole alkanols II can be prepared by methods, such as those de,~cribed by F. Schneider, Z.
Physiol . Chem ., 3 :206-210 (1961) and C.P. Stewart, Biochem. Jou~ nal, 17:130-133(1923). Benzylation and deprotection of the imidazole alkanol provides an intermediate which can be oxidized to the corresponding aldehyde.
The aldehyde IV is reacted with a suitably substituted amine. The interrnediate can then be reacted with a sulfinyl chloride.
Syntheses of suitably sub.stituted aldehydes are illustrated wherein the variable W is present as a pyridyl moiety. Similar synthetic strategies for preparing alkanols that incorporate other heterocyclic moieties for variable W are also well known in the art.
The ~sulfinamide can be formed by converting a hydroxyl group to a .sulfinyl chloride, and then reacting the sulfinyl chloride with the appropriately ~substituted amine. The re.sulting .sulfinamide can thereafter be oxidized with, e.g., periodate, to produce sulfonamides in accordance with fonnula I. Likewi~e, by reacting the precur.sor amine with an alkylating agent, .~ubstitution on the sulfonamide nitrogen atom can be realized.

(CR22)pl-cH2oH Prot1X Et3N Prot 'N~/~>

H DMF lla (CR22)p,-CH20Ac R6~EtOAc AC20, Py ,=1 Prot1~N~ N 2. N-deprotect ~CR 2)p-CH20Ac N~'~(CR 2)p-CH20H

N~ LiOH >
R6~ THF, H20 ~

~(CR 2)p-CHO (CR22)p-CH2NHCH3 SO3 Py, Et3N ,=~) CH3NH2 N

- DMSO ~ Na(AcO)3B~

IV V

OH
DEAD ~ SC(O)CH3 [~PPh3 CH3C(O)SH

Ac20 /S(O)CI Et~N
CH2CI2 ¢~ CH2CI2 H3CHN ~~

(R )r--N(Me)S(O) R4 ~SO _ NalO4 ~N~ R 4 (R )r ~ RuCI3 ~ R

~ HCHO, aq.
R3 R4 CH3CN, NaCNBH3 ~/\NHCH3 OH
SC(O)CH3 DEAD ~\N
6)~ PPh3 CH3C(O) S(O)CI

Ac20 N~N Et~N

Cocc2 ~ R3 S(O)NH~

R NalO4 ~' ~ R4 ~ RoCI3 (R6)r/~J aq.

(R6)r R7 CH3 1) HNO2,Br2 7 ~CO2CH3 ~ 2)KMnO
H2N N~ 3) MeOH,H+ Br N

R~MgCI R\ ~\5' CO2CH3 ZnCI2,NiCl2(Ph3p)2 N

NaBH4 (excess) ~,CH20H

S03 Py Et3N ~ CHO

CH3NH2 ~\~ \~CH2NHCH3 Na(AcO)~BH ~ ~ J

~\~Co2cH3 ~\ 9 ~"Co2cH3 Zn, CuCN

NaBH4 ~j~ SO3Py, Et3N ~
(excess) R~\~CH20H DMSO R\~CHO

Br~C02cH3 I~\MgCI ~ C02CH3 N ZnCI2, NiCI2(Ph3P)2 R7 ~

NaBH4 ¦ SO3Py, Et3N
~ CH20H ~ ~ CHO
(excess) ~N ~ DMSO ~'N

CH3NH2 ¢~

Na(AcO)3BH ~,CH2NHCH3 Br~1. LDA, CO2 Br~

N2. MeOH, H+ N

~/\MgCI ~ CH3 ZnCI2, Nicl2(ph3p)2 N~

NaBH4 (excess) ~ CH20H SO3 Py, Et3N

~ R7 DMSO

CHO CH3NH2 ~ CH2NHCH3 Na(AcO)3BH ~? - R7 R7 ~' 1. LDA, CO2 Br 2. (CH3)3SiCHN2 R6 ~\Br R6 ~

N ~ C02C H3 Zn, NiCI2(Ph3P)2 R7 ;~

R6 1~
excess NaBH4 l~ SO3 Py, Et3N
R7 N;~CH20H DMSO

R6 ~ CH3NH2 N~,CHO Na(AcO)3BH 7 N~

WO 97/36583 PCTtUS97tO5170 Boc NHl IX CH3NH2 HCI Boc NHl Boc NH CHO NaBH(OAc)3 Boc NH CH2NHCH3 Et3N . CICH2CH2CI

/ ~
J

B~CNHl f/'' 3 Boc NH~NcH3s(o3~

RuCI3\CF3CO2H
aq.\(~,H2CI2 Boc NH~ 1~--R3 Boc NH NCH3S02 R4 R4 CF3CO2H H2N ~ R3 CH2C12 H2N NCH3S(Ot /

~, R4 H2N ~ ~ R3 BnOl CH3Nlt2-HCI

Boc NH CHONaBH(OAc)3 Et3N, CICH2CH2CI

BnO ~ Et3N

Boc NH CH2NHCH3 S(O)CI

~1 R 4 Nal~4 BnO ~ R3 RuCI3 Boc NH NCH3S(O

.. .... . . ..

H2N ~ ~/~/--R3 Boc20 R4 ,~CH0 BocHN ~ ~ R3 l~J

H2N NCH3S02 NaBH(OAc)3 Et3N, CICH2CH2CI

BocHN ~ 3 ~J CF3CO2H, CH2CI2 H2N~ R3 ~NC

AgCN
,~

W <N~ R3 -2~ -~4 BnO ~ ~R 20% Pd(~H)2~ H2 ~ ~ CH30H
Boc NH NCH3S02 CH3CO2H

HO ~/~ R3 ') CICOCOCI
Boc NH --NCH3S02--~/ DMSO CH2C12 (C2H5)3N

R4 1. R'MgX
H~O ~/~R3 (C2Hs)2~

~ 2 TFA
Boc NH NCH3S02 CH2C12 ,~4 R, OH ~ ~R3 H0~ ~ R3 CF3CO2H
Boc NH NCH3S02 CH2CI2 H0 ~, R R'CH0 H2N~--NCH3S0-- NaBH(OAc)3 H0 ~/';\ R3 R'C H2N H NC H3S02 N
R4 N=\N~ ,N/=
HO~ ~/~ R3 ~ O

Boc NH NCH3S02 NaH, DMF 0~C

~/~R3 R'SH
~ (c2H5)3 Boc N --NCH3S02 CH30H

R'S ~ ~ HCI, EtOAc Boc NH NCH3S02 R'S~ ~R3 HO~ 1) Boc20, K2C~3 HO~

,~/ THF-H20 ~
2) C H2N EtOAc H2N CO2H 2, BocNH CO2CH3 XXVI I
XXVIII

HO~
LiAlH4 ~JJ R'CH2X
THF ~ Cs2CO3 0-20~C BocNH CH2OH DMF

XXIX

R 'C H20~ R 'C H20~

,~W OMSO ,~lV
BocNH CH2OH (C2Hs)3N BocNH CHO
20~C
XXX XXXI

R'CH ~3 CH3NH2-HCI RCH20 Na(OAc)3BH
BocNH CHO CICH2CH2CI BocNH/~ NHCH3 ~S(O)CI

R'CH

BocNH NCH3S(O
NalO4\
RuCI3 \~
aq.
R'CH~,,~ 3 ~R4 BocNH NCH3s02 ~/~ 20% Pd(OH)2 \
CH30H, CH3C02H\ EHtCOA
2. HCI, EtOAC
R'CH~O~ r~,R4 CA 022~0143 1998-09-2~

The instant compounds are useful in the treatment of cancer. Cancer.s which may be treated with the compound,s of thi~
invention include, but are not limited to, colorectal carcinoma, exocrine pancreatic carcinoma, myeloid leukemia,s and neurological tumors. Such tumors may arise by mutation~ in the ra~ genes themselves, mutation.s in the proteins that can l-egulate Ra,s activity (i.e., neurofibromin (NF-l), neu, ,scr, abl, Ick, fyn) or by other mechanism,~.
The compound,~i of the in~itant invention inhibit farne,syl-protein tr~n~sferase and tarne~ylation of the oncogene protein Ra,s.
The instant compounds may al,so inhibit tumor angiogenelsi,~, thereby affecting the growth of tumors (J. Rak et al. Cfln( e~- Re~earch, 55:4575-45~0 (1995)). Such anti-~ngiogenic propertie,s of the in~tant compounds may also be u~eful in the treatment of certain form~s of blindnes,<i related to retinal vascularization.
The compounds of this invention are also u,seful for inhibiting other diseases where Ras protein,s are aberrantly activated as a result of oncogenic mutation in other gene,s (i.e., the Ra,s gene itself i~
not activated by mutation to an oncogenic form) with said inhibition being accomplished by the administration of an effective amount of the compound,s of the invention to a mammal in need of ,such treatment.
For example, a component of NF-I is a benign proliferative disorder.
The instant compounds may also be u~eful in the treatment of viral infections, in particular in the treatment of hepatiti,s delta and related viru,se,s (J.S. Glenn et al. Scienef~, 256: 1331 - 1333 (1992).
The compounds of the in~tant invention are al,so useful in the prevention of resteno,~ after percutaneou,s tran,sluminal coronary angioplasty by inhibiting neointimal formation (C. Indolfi et al. Natu~ e medicine, 1 :541-545(1995).
The instant compounds may al~o be useful in the treatment and prevention of polycy,stic kidney disease (D.L. Schaffner et al.
American J(~urnal (~f Path(~lo(~y~ 142:1051-1060 (1993) and B. Cowley, Jr. et al.FASEB Jf~urnal, 2:A3160 (19~

CA 022~0143 1998-09-2~

The instant compounds may also be useful for the treatment of fungal infections.
The compounds of this invention may be administered to m;lmm~l~;, preferably humans, either alone or, preferably, in combina-S tion with pharmaceutically acceptable carriers or diluents, in the form of a pharmaceutical composition, which is comprised of a compound of formula I in combination with a pharmaceutically acceptable carrier.
The compound.s can be admini~stered orally, topically, rectally, vaginally transdermally or parenterally, including the intravenous, intramuscular, 10 intraperitoneal and subcutaneous routes of administration.
For oral use, the compound is administered, for example, in the form of tablets or capsules, or as a solution or ~suspension. ln the case of tablets for oral use, carrier~s which are commonly used include lactose and corn starch; lubricating agents, such as magnesium stearate, 15 are commonly added. For~ral administration in capsule form, diluents also include lactose and dried corn starch. When a4ueou.s suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring agents may be added. For intramuscular, intra-20 peritoneal, subcutaneous and intravenous use, sterile solution~s of the active ingredient are usually prepared, the pH of the solution is ~suitably adjusted and the product is buffered. For intravenous use, the total concentration is controlled to render the preparation substantially isotonic.
The compounds of the instant invention may also be co-:~(lministered with other well known therapeutic agents that are selected for their particular usefulness against the condition that is being treated. For example, the instant compounds may be useful in combination with known anti-cancer and cytotoxic agents. Similarly, the instant compounds may be useful in combination with agents that are effective in the treatment and prevention of NF-I, restinosis, polycystic kidney disease, infections of hepatitis delta and related viruses and fungal infections.

CA 022~0143 1998-09-2~

WO 97t36583 PCT/US97/05170 If formulated as a fixed dose, such combination product,s employ a compound of this invention substantially within the dosage range de,scribed below and other pharmaceutically active agent(s) typically within the acceptable do,sage range. Compounds of the instant S invention may alternatively be u,sed se(Juentially with known pharma-ceutically acceptable agent(,s) when a combination forrnulation is inappropriate.
The daily dosage will normally be determined by the prescribing physician, who may vary the do,sage according to the age, 10 weight, and respon,se of the individual patient, as well as the severity of the patient'~s condition.
In one exemplary application, a suitable amount of compound i.s administered to a m~mm~l undergoing treatment for cancer. Admini~stration occurs in an amount between about 0.1 mg/kg 15 of body weight to about 60 mg/kg of body weight per day, preferably of between 0.5 mg/kg of body weight to about 40 mg/kg of body weight per day.
The compound,s of the instant invention are also u~seful as a component in an assay to rapidly determine the pre~sence and 20 quantity of farne.syl-protein transferase (FPTa,se) in a composition.
Thu.s the composition to be tested may be divided and the two portions contacted with mixtures which comprise a known ,substrate of FPTase (for example a tetrapeptide having a cysteine at the amine terminus) and farnelsyl pyrophosphate and, in one of the mixture~s, 25 a compound of the instant invention. After the assay mixtures are incubated for an ,sufficient period of time, well known in the art, to allow the FPTase to farnesylate the substrate, the chemical content of the assay mixture,s may be determined by well known immuno-logical, radiochemical or chromato~raphic technique~. Because 30 the compounds of the instant invention are selective inhibitors of FPTa~se, ab.sence or quantitative reduction of the amount of substrate in the a~s~say mixture without the compound of the instant invention relative to the prelsence of the unchanged ~sub~strate in the assay CA 022~0143 1998-09-2~

containing the instant compound is indicative of the presence of FPTa,se in the composition to be tested.
It would be readily apparent to one of ordinary skill in the art that ~uch an assay as described above would be useful in identifying 5 tissue sample,s which contain farne~yl-protein tran~ferase and quanti-tating the enzyme. Thus, potent inhibitor compounds of the instant invention may be used in an active site titration assay to determine the quantity of enzyme in the sample. A ~erie~ of !;ample~i composed of aliquots of a tissue extract containing an unknown amount of farne,syl-10 protein tran~sferase, an exces~; amount of a known substrate of FPTa~ie (for example a tetrapeptide havin~ a cy~teine at the amine terrninus) and farnesyl pyrophosphate are incubated for an appropriate period of time in the presence of varying concentrations of a compound of the instant invention. The concentration of a sufficiently potent inhibitor (i.e., one lS that ha.s a Ki substantially ~smaller than the concentration of enzyme in the as~;ay vessel) required to inhibit the enzymatic activity of the sample by 50% is approximately equal to half of the concentration of the enzyme in that particular sample.

N~ (4-CYANOBENZYL)IMIDAZOLYL-5-METHYLl-N-(METHYL)-3 -CHLOROBENZYLSULFONAMIDE
HYDROCHLORIDE
Step A: Preparation of l-triphenylmethyl-4-~hydroxymethyl)imidazole To a .~olution of 4-(hydroxymethyl)imidazole hydrochloride (35.0 g, 260 mmol) in 250 mL of dry DMF at room temperature wa~ added triethylamine (90.6 mL, 6~0 mmol). A white 30 solid precipitated from the solution. Chlorotriphenylmethane (76.1 ~, 273 mmol) in 500 mL of DMF wa~i added dropwi~ie. The reaction mixture wa.s ,stirred for 20 hour~s, poured over ice, filtered, and wa~;hed with ice water. The resulting product wa.~ ~lurried with cold dioxane, filtered, and dried i)1 V~ 'UO to provide the titled product a~ ~ white solid 35 which wa~ I~ufficiently pure for u~se in the next .step.

CA 022~0143 1998-09-2~

Step B: Preparation of 1 -triphenylmethyl-4-(acetoxvmethyl)imidazole Alcohol from Step A (260 mmol, prepared above) wa~
su,spended in 500 mL of pyridine. Acetic anhydride (74 mL, 780 S mmol) wa,s added dropwi.se, and the reaction wa,s ,stirred for 4~ hours during which it became homogeneous. The solution was poured into 2 L of EtOAc, washed with water (3 x 1 L), 5% a~. HCl ,soln. (2 x 1 L), ,sat. aq. NaHCO3, and brine, then dried (Na2SO4), filtered, and concentrated in va~o to provide the crude product. The acetate wa~
10 ilsolated as a white powder (g5.X g, ~6% yield for two .step.s) which wa~
.sufficiently pure for use in the next reaction.

Step C: Preparation of 1-(4-cyanobenzyl)-5-(acetoxymethyl)imidazole hydrobromide A ~solution of the product from Step B (~5.X g, 225 mmol) and ~-bromo-p-tolunitrile (50.1 g, 232 mmol) in 500 mL of EtOAc wa~i stirred at 60~C for 20 hour~s, during which a pale yellow precipitate formed. The reaction wa,s cooled to room temperature and filtered to provide the ~solid imidazolium bromide ,salt. The filtrate wa,s 20 concentrated in vac~u~ to a volume 200 mL, reheated at 60~C for two hour.s, cooled to room temperature, and filtered again. The filtrate was concentrated in vac~uo to a volume 100 mL, reheated at 60~C for another two hours, cooled to room temperature, and concentrated in vacuo to provide a pale yellow solid. All of the Isolid material was 25 combined, dissolved in 500 mL of methanol, and walmed to 60 ~C.
After two hours, the solution wa,s reconcentrated in vacuo to provide a white lsolid which wa,s triturated with hexane to remove .soluble material,s. Removal of re,sidual ,solvents il~ vacu~ provided the titled product hydrobromide a~s a white solid (50.4 g, 67~/~, yield, ~9% purity 30 by HPLC) which wa~ u,sed in the next ,step without further purification.

Step D: Preparation of 1-(4-cyanobenzyl)-5-(hydroxymethvl)imidazole To a ~;olution of the acetate from Step C (50.4 ~, 150 mmol) in 1.5 L of 3:1 THF/water at 0 ~C wa,s added lithium hydroxide CA 022~0143 1998-09-2~

WO 97/36583 PCTtUS97/0~170 -3g-monohydrate (18.9 g, 450 mmol). After one hour, the reaction was concentrated in l~acuo, diluted with EtOAc (3 L), and washed with water, sat. aq. NaHCO3 and brine. The solution was then dried (Na2SO4), filtered, and concentrated in vacuo to provide the crude product (26.2 g, 82% yield) as a pale yellow fluffy solid which was sufficiently pure for use in the next .step without further purification.

Step E: Preparation of 1-(4-cyanobenzyl)-5-imidazolecarboxaldehyde To a solution of the alcohol from Step D (21.5 g, 101 mmol) in 500 mL of DMSO at room temperature was added triethyl-amine (56 mL, 402 mmol), then SO3-pyridine complex (40.5 g, 254 mmol). After 45 minutes, the reaction was poured into 2.5 L of EtOAc, washed with water (4 x 1 L) and brine, dried (Na2SO4), filtered, and concentrated in vacMo to provide the aldehyde (1 ~.7 g, ~X% yield) as a white powder which was sufficiently pure for use in the next step without further purification.

Step F: Preparation of 1-(4-cyanobenzyl)-5-I (methylamino)methyllimidazole To a suspension of methylamine hyd~ochloride in 5 mL
dichloroethane at 0~C are added 4 A sieves (0.5g), followed by 1 mmol of the aldehyde from Step E and 1.5 mmol Na(OAc)3BH. The reaction is stirred for 10 minutes at 0~C, warmed to room temperature and stirred for 2 hours. The reaction i~s poured into EtOAc/ ~sat. NaHCO3 solution. The organic layer i~s washed with brine, dried (Na2SO4 ), filtered, and concentrated in vacuo. The crude product is taken up in 10 mL dichloromethane and 2 mL n-propylamine, stirred at room temperature for I hour, concentrated in vacuo, and purified by flash chromatography .
Step G: Preparation of 3-chlorobenzyl thioacetate To a solution of 1 mmol 3-chlorobenzyl alcohol and I mmol triphenylpholsphine in 5 mL THF at 0~ C, I mmol diethyl-CA 022~0143 1998-09-2~

WO 97/36583 PCT/U~97/05170 azodicarboxylate is added. After stirring for 10 minutes, I mmol thioacetic acid is added. The reaction i~s stirred for 3 hours, concentrated and purified by flash chromatography.

StepH: Preparation of 3-chlorobenzyl.sulfinyl ch~oride To a -20~ C solution of the thioester from Step G (1 mmol) in 3 mL of dichloromethane under ar~on are added I mmol acetic anhydride and 2 mmol sulfuryl chloride. The reaction is stirred for I
hr during which time the temperature is allowed to rise to -5~ C. The mixture is concentrated in ~~ac~uo and the crude product used for coupling in the next step.

Step I: Preparation of N-[1-(4-cyanobenzyl)imidazolyl-~-methyl]-N-(methyl)-3 -chlorobenzylsulfinamide To a 0~ C solution of the sulfinyl chloride from Step H
(I mmol) in 4 mL dichloromethane i,s added a solution of 1 mmol triethylamine and I mmol of the amine from Step F in 2 mL dichloro-methane. The reaction i,~ stirred overnight and the temperature is allowed to rise to room temperature. This mixture is poured into ethyl acetate, washed with sat. NaHCO3 solution and brine, dried with Na2SO4, concentrated and purified by fla,sh chromatography.

Step J: Preparation of N-[ 1 -(4-cyanobenzyl)imidazolyl-5-methyl] -N-(methyl)-3-chlorobenzylsulfonamide hydrochloride A solution of the ,sulfinamide from Step I (1 mmol) in 2 mL acetonitrile is cooled to 0~C. NaIO4 i,~i added (1.5 mmol) followed by a catalytic amount of RuCI~ 3H~O and 2 mL H2O. The reaction is stirred at room temperature for 1 hour, diluted with EtOAc, and washed with ,sat. NaHCO3 solution and brine, dried (Na2SO4), filtered and concentrated. The resulting product is purified by flash chromatography~ then taken up in dichloromethane, and treated with with excess ethereal HCI. Concentration in ~ UO provides the titled product.

WO 97/36583 PCT/US97/0~170 N-(3-CHLOROBENZYL)-1 -I (4-CYANOBENZYL)-5-IMIDAZOLYLlMETHYLSULFONAMIDE HYDROCHLORIDE

Step A: Preparation of 1-(4-chlorobenzyl)-5-imidazolylmethyl sulfinyl chloride The titled compound i.s prepared from the alcohol from Step D of Example 1 by the method.s described in Steps G and H of Example 1.

Step B: Preparation of N-(3-chlorobenzyl)-1-L(4-cyanobenzyl)-5-imidazolyllmethylsulfinamide The sulinyl chloride from Step A i.s coupled with 3-chlorobenzylamine using the method de~icribed in Step I of Example 1.

Step C: Preparation of N-(3-chlorobenzyl)- 1 -[(4-cyanobenzyl)-5-imidazolyllmethylsulfonamide hydrochloride The sulfinamide from Step B is oxidized to the sulfonamide and converted to the HCI ~alt using the method delscribed in Step J of Example 1.

N-l 3-(4-CYANOBENZYL)PYRIDYL-4-METHYLl-N-(METHYL)-3-CHLOROBENZYLSULFONAMIDE HYDROCHLORIDE

Step A: Preparation of 3-(4-cyanobenzyl)pyridin-4-carboxylic acid methyl ester A solution of 4-cyanobenzyl bromide (625 mg, 3.27 mmol) in dry THF (4mL) wa,s added .slowly over~3 min. to a suspension of activated Zn (dust; 250 mg) in dry THF (2 mL) at O~ under an argon atmosphere. The ice-bath wa~s removed and the ,slurry wa~i ,stirred at room temperature for a further 30 min. Then 3-bromopyridin-4-carboxylic acid methyl ester (540 mg. 2.5 mmol) followed by CA 022~0143 1998-09-2~

dichlorobis(triphenylphosphine)nickel (II) (50 mg). The resultant reddish-brown mixture was stirred for 3h at ~40-45~C. The mixture was cooled and distributed between EtOAc (100 ml) and 5% a4ueous citric acid (50 mL). The organic layer was washed with H2O (2XS0 5 mL), dried with Na2SO4. After evaporation of the solvent the residue was purified on silica gel, eluting with 35% EtOAc in hexane to give 420 mg as a clear gum. FAB ms (M+l) 253.

Step B: Preparation of 3-(4-cyanobenzyl)-4-(hydroxymethyl)pyridine The title compound was obtained by ~odium borohydride (300 mg) reduction of the ester from Step A (41~ mg) in methanol (5 mL) at room temperature. After stirring for 4 h the .~;olution was evaporated and the product wa~i purified on silica gel, eluting with 2%
methanol in chloroform to give the title compound. FAB m~; (M+l) 1 5 225.

Step C: Preparation of 3-(4-cyanobenzyl)-4-pyridinal The title compound was obtained by activated manganese dioxide (l.Og) oxidation of the alcohol from Step B (240 mg, 1.07 mmol) in dioxane (10 mL) at reflux for 30 mim Filtration and evaporation of the ~olvent provided title compound, mp ~0-~3~C.

Step D: Preparation of 3-(4-cyanobenzyl)-4-I (meth ylamino)methyl lpyridine The titled compound i~; prepared from the pyridinal from Step C and methylamine hydrochloride using the procedure in Step F of Example 1.

Step E: Preparation of N-[3-(4-cyanobenzyl)pyridyl-4-methyl]-N-(methyl)-3-chlorobenzyl~ulfon~mide hydrochloride The titled compound i~ prepared from the amine from Step D and the sulfinyl chloride from Step H of Example I u~iing the procedure.s de.scribed in Step.~; I and J of Example 1.

CA 022~0143 1998-09-2~

ln vitro inhibition of ra.s farnesyl tran~sferase Assays of farnesyl-protein t~ aMsfe7 ase~ Partially purified bovine FPTase and Ras peptides (Ras-CVLS, Ras-CVIM and Ras-CAIL) were prepared as described by Schaber et al., J. Biol. Chem. 265:14701-14704 (1990), Pompliano, et al., Biochemistry 31:3~00 (1992) and Gibbs et ah, PNAS U.S.A. ~6:6630-6634 (1989), respectively. Bovine FPTa,se was assayed in a volume of 100 ~I cont~ining 100 mM N-(2-hydroxy ethyl) piperazine-N'-(2-ethane .sulfonic acid) (HEPES), pH
7.4, 5 mM MgC12, 5 mM dithiothreitol (DTT), 100 mM ~3H]-farne.syl diphosphate ([3H]-FPP; 740 CB4/mmol, New England Nuclear), 650 nM
Ras-CVLS and 10 ,ug/ml FPTase at 31 ~C for 60 min. Reactions were initiated with FPTa.se and stopped with 1 ml of 1.0 M HCL in ethanol.
Precipitates were collected onto filter-mat~; using a TomTec Mach II cell harvestor, washed with 100% ethanol, dried and counted in an LKB ,B-plate counter. The assay wals linear with re.spect to both substrates, FPTase levels and time; less than 10% of the [3H]-FPP wa.s utilized during the reaction period. Purified compound~ were dis.solved in 100% dimethyl sulfoxide (DMSO) and were diluted 20-fold into the assay. Percentage inhibition is mea.sured by the amount of incorporation of radioactivity in the presence of the te.st compound when compared to the amount of incorporation in the ab.sence of the test compound.
Human FPTase was prepared as described by Omer et al., Biochemistry 32:5167-5~76 (1993). Human FPTase activity was assayed as described above with the exception that 0.1 % (w/v) polyethylene glycol 20,000, 10 ~lM ZnC12 and 100 nM Ras-CVIM were added to the reaction mixture. Reactions were performed for 30 min., stopped with 100 ~1 of 30% (v/v) trichloroacetic acid (TCA) in ethanol and processed a.s described above for the bovine enzyme.
n vil~o ra.s farne!iylation assay The cell line used in this a.ssay is a v-ras line derived from either Ratl or NIH3T3 cells, which expressed viral Ha-ras p21.
The assay is performed e~sentially as de.scribed in DeClue, J.E. et al., CA 022~0143 1998-09-2~

Cancer Research 51 :712-717, (1991). Cells in 10 cm dishe.s at 50-75%
confluency are treated with the te,st compound (final concentration of ~solvent, methanol or dimethyl sulfoxide, is 0.1%). After 4 hours at 37~C, the cells are labelled in 3 ml methionine-free DMEM supple-5 meted with 10% regular DMEM, 2% fetal bovine serum and 400mCi[35S]methionine (1000 Ci/mmol). After an additional 20 hours, the cells are Iysed in 1 ml Iy.sis buffer (1% NP40/20 mM HEPES, pH 7.5/5 mM MgC12/lmM DTT/10 mg/ml aprotinen/2 mg/ml leupeptin/2 mg/ml antipain/0.5 mM PMSF) arld the Iysates cleared by centrifugation at 100,000 x g for 45 min. Aliquot.s of Iysate,s containing equal numbers of acid-precipitable count.s are bought to 1 ml with IP buffer (Iysis buffer lacking DTT) and immunoprecipitated with the ras-specific monoclonal antibody Y13-259 (Furth, M.E. et al., J. Virol. 43:294-304, (19~2)). Following a 2 hour antibody incubation at 4~C, 200 ml of a 15 25% suspension of protein A-Sepharo.se coated with rabbit anti rat IgG
is added for 45 min. The immunoprecipitate,s are wa~shed four times with IP buffer (20 nM HEPES, pH 7.5/1 mM EDTA/I % Triton X-100Ø5% deoxycholate/0.1%/SDS/0.1 M NaCl) boiled in SDS-PAGE
,sample buffer and loaded on 13% acrylamide gels. When the dye front 20 reached the bottom, the gel is fixed, soaked in Enlightening, dried and autoradiographed. The intensities of the band,s corre,sponding to farne,sylated and nonfarnesylated ra,s proteins are compared to determine the percent inhibition of farnesyl tran,sfer to protein.

25 In Vil)O ~rowth inhibition assay To deterrnine the biological consequences of FPTase inhibition, the effect of the compounds of the instant invention on the anchorage-independent growth of Ratl cell,s transformed with either a v-ras, v-7 af, or v-mos oncogene is te,sted. Cells transformed by v-Raf 30 and v-Mos maybe included in the analysi,s to evaluate the ,specificity of instant compounds for Ras-induced cell transformation.
Rat 1 cells tran,sformed with either v-ras, v-raf, or v-mo~
are ~eeded at a density of 1 x 104 cell.s per plate (3~ mm in diameter) in a 0.3% top agarose layer in medium A (Dulbecco'~ modified Eagle's medium supplemented with 10% fetal bovine .serum) over a bottom agaro~ie layer (0.6%). Both layers contain 0.1% methanol or an appropriate concentration of the in.stant compound (dissolved in methanol at 1000 times the final concentration used in the assay). The S cells are fed twice weekly with 05 ml of medium A containing 0.1%
methanol or the concentration of the instant compound. Photomicro-graphs are taken 16 days after the cultures are seeded and comparisons are made.

Claims (32)

WHAT IS CLAIMED IS:

1. A compound represented by formula I:

or a pharmaceutically acceptable salt thereof. wherein:

R1a, R1b and R2 are independently selected from the group consisting of: hydrogen, aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(O)m-, (R8)2NC(O)-, R8C(O)NR8-, CN, NO2, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(O)NR8- and C1-C6 alkyl, unsubstituted or substituted by 1-3 groups selected from the group consisting of: halo, aryl. heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(O)m-, R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8sOC(O)-, N3, -N(R8)2 and R9OC(O)NR8-;

R3 and R4 are independently selected from the group consisting of: H, F, Cl, Br, -NR8 2, CF3, NO2, R8O-, R9S(O)m-, (R8)2NC(O)-, R8C(O)NH-, H2NC(NH)-, R8C(O)-, R8OC(O)-, N3, CN, R9OC(O)NR8-, C1-C20 alkyl, substituted or unsubstituted aryl and substituted or unsubstituted heterocyclyl;

A3 is selected from: -NR5S(O)m- or -S(O)mNR5-, with m equal to 0, 1 or 2, and R5 selected from the group consisting of:
hydrogen, unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted C3-C10 cycloalkyl, and C1-C6 alkyl, unsubstituted or substituted with 1-3 members selected from the group consisting of: unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted C3-C10 cycloalkyl, -N(R8)2, -CF3, -NO2, (R8)O-, (R9)S(O)m-, (R8)C(O)NH-, H2NC(NH)-, (R8)C(O)-, (R8)OC(O)-, N3, CN and (R9)OC(O)NR8-;

R6 and R7 are independently .selected from the group consisting of: hydrogen, aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-6 perfluoroalkyl, F, Cl, Br, R8O-, R9S(O)m-, R8C(O)NR8-, CN, NO2, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(o)NR8- and C1-C6 alkyl unsubstituted or substituted by 1-3 groups .selected from: aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R8O-, R9S(O)m-, R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2 and R9OC(o)NR8-;

each R8 is independently selected from hydrogen, C1-C6 alkyl, aryl and aralkyl;

each R9 is independently selected from C1-C6 alkyl and aryl;

A1 and A2 are independently selected from the group consisting of: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR8-, -NR8C(O)-, -O-, -N(R8)-, -S(O)2N(R8)-, -N(R8)S(O)2-, and S(O)m;
X represents aryl or heteroaryl;

V is selected from the group consisting of: hydrogen, heterocyclyl, aryl, C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S. and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond, n is 0 and A2 is S(O)m;
W represents heterocyclyl;

each n and p independently represents 0, 1, 2, 3 or 4;
r is 0 to 5, provided that r is 0 when V is hydrogen, and t is 0 or 1.

2. A compound in accordance with claim 1 wherein R1a, R1b and R2 are independently selected from the group consisting of: hydrogen, -N(R8)2, R8C(O)NR8- and C1-6 alkyl, unsubstituted or substituted with 1-2 groups selected from unsubstituted or substituted aryl, -N(R8)2, R8O- and R8C(O)NR8- .

3. A compound in accordance with claim 1 wherein R3 and R4 are selected from: hydrogen and C1-C6 alkyl.

4. A compound in accordance with claim 1 wherein A3 represents NR5S(O)m, in which m represents 2 and R5 represents hydrogen.

5. A compound in accordance with claim 1 wherein R6 represents hydrogen, unsubstituted or substituted C1-C6 alkyl.

6. A compound in accordance with claim 1 wherein R7 represents H or unsubstituted C1-6 alkyl.

7. A compound in accordance with claim 1 wherein R8 represents H or C1-6 alkyl, and R9 is C1-6 alkyl.

8. A compound in accordance with claim 1 wherein A1 and A2 are independently selected from: a bond, -C(O)NR8-, -NR8C(O)-, -O-, -N(R8)-, -S(O)2N(R8)- and-N(R8)S(O)2-.

9. A compound in accordance with claim 1 wherein V is selected from hydrogen heterocyclyl and aryl.

10. A compound in accordance with claim 1 wherein V
represents phenyl.

11. A compound in accordance with claim 1 wherein X
represents aryl.

12. A compound in accordance with claim 11 wherein X
represents phenyl.

13. A compound in accordance with claim I wherein W
is heterocyclyl selected from the group consisting of: imidazolinyl, imidazolyl. oxazolyl, pyrazolyl, pyyrolidinyl, thiazolyl and pyridyl.

14. A compound in accordance with claim 10 wherein W
is selected from imidazolyl and pyridyl.

15. A compound in accordance with claim 1 wherein m is 0 or 2.

16. A compound in accordance with claim 1 wherein n and p are 0, 1, 2 or 3.

17. A compound in accordance with claim 1 wherein t is 1.

18. A compound in accordance with claim 1 represented by formula Ia:

wherein:

R3, R4, A3, R8, R9, m, n, p and r are as originally defined;

each R1a and R2 is independently selected from hydrogen and C1-C6 alkyl;
each R1b is independently selected from: hydrogen, aryl, heterocyclyl, C3-10 cycloalkyl, C2-6 alkenyl, R8O-, -N(R8)2 and C1-C6 alkyl unsubstituted or substituted by aryl, heterocyclyl, cycloalkyl, alkenyl, R8O- and -N(R8)2;

R5 is selected from the group consisting of: hydrogen and C1-C6 alkyl, unsubstituted or substituted with 1-3 members selected from the group consisting of: unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted C3-C10 cycloalkyl, -N(R8)2, -CF3, -NO2, (R8)O-, (R9)S(O)m-, (R8)C(O)NH-, H2NC(NH)-, (R8)C(O)-, (R8)OC(O)-, N3, CN and (R9)OC(O)NR8-;

R6 is independently selected from: hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F. Cl, R8O-, R8C(O)NR8-, CN, NO2, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(RX)2, or R9OC(O)NR8-, and C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R8O-, R8C(O)NR8-, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2 and R9OC(O)NR8-;

R7 is H or unsubstituted C1-6 alkyl;

A1 and A2 are independently selected from: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR8-, O, -N(R8)- and S(O)m;

and V is selected from: hydrogen; aryl; heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl and thienyl; C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond and A2 is S(O)m.

19. A compound in accordance with claim 1 represented by formula Ib:

wherein:

R1a, R1b, R2, A1, A2, R3, R4, R5, R6, R8, R9, m, n, p and r are as originally defined;

R7 is selected from: hydrogen and unsubstituted C1-C6 alkyl;

V is selected from: hydrogen, heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, aryl, C1-C20 alkyl wherein from O to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond, n is 0 and A2 is S(O)m; and W represents heterocyclyl selected from pyrrolidinyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl and isoquinolinyl.

20. A compound in accordance with claim 1 represented by formula Ic:

wherein:

each R2 is independently selected from hydrogen and C1-C6 alkyl;
R3, R4, A3, R8, R9, m and p are as originally defined;

each R5 is selected from: hydrogen and C1-C6 alkyl unsubstituted or substituted with 1-3 groups selected from unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted C3-C10 cycloalkyl, -N(R8)2, -CF3, -NO2, (R8)O-, (R9)S(O)m-, (R8)C(O)NH-, H2NC(NH)-, (R8)C(O)-, (R8)OC(O)-, N3, -CN and (R9)OC(O)NR8-;

and R6 is selected from the group consisting of: hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R8O-, R8C(O)NR8-, CN, NO2, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2, or R9OC(O)NR8- and C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R8O-, R8C(O)NR8-, (R8)2N-C(NR8)-, R8C(O), R8OC(O)-, -N(R8)2 or R9OC(O)NR8-.

21. A compound in accordance with claim 1 represented by formula Id:

wherein:
each R2 is independently selected from: hydrogen and C1-C6 alkyl;

R3 and R4 are independently selected from H, F, Cl, Br, N(R8)2, CF3, NO2, (R8)O-, (R9)S(O)m-, (R8)C(O)NH-, H2N-C(NH)-, (R8)C(O)-, (R8)OC(O)-, N3, CN, (R9)OC(O)NR8-, C1-C20 alkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclyl;
A3 represents -NR5-S(O)m- or -S(O)m-NR5-;

R5 is selected from: hydrogen and C1-C6 alkyl, unsubstituted or substituted witha group selected from unsubstituted or substituted aryl, unsubstituted or substituted heterocyclyl, unsubstituted or substituted C3-C10 cycloalkyl, N(R8)2, CF3, NO2, (R8)O-, (R9)S(O)m-, (R8)C(O)NH-, H2N-C(NH)-, (R8)C(O)-, (R8)OC(O)-, N3, CN (R9)OC(O)NR8-;
and R8, R9, m and p are as originally defined.

22. A compound in accordance with claim 1 represented by the formula:

or or a pharmaceutically acceptable salt thereof.

23. A pharmaceutical composition which comprises a compound in accordance with claim 1 in combination with a pharmaceutically acceptable carrier.

24. A method of inhibiting farnesyl-protein transferase in a mammalian patient in need of such treatment which comprises administering to said mammal a compound in accordance with claim 1.

25. A method of treating cancer in a mammalian patient in need of such treatment which comprises administering to said patient an anti-cancer effective amount of a compound in accordance with claim 1.

26. A method of treating cancer in accordance with claim 23, wherein the cancer treated is selected from colorectal carcinoma, exocrine pancreatic carcinoma, myeloid leukemia and neurological tumors.

27. A method of treating neurofibromin benign proliferative disorder in a mammalian patient in need of such treatment which comprises administering to said patient an effective amount of a compound of Claim 1.

28. A method of treating blindness related to retinal vascularization in a mammalian patient in need of such treatment which comprises administering to said patient an amount of a compound of claim 1 which is effective for treating blindness related to retinal vascularization.

29. A method of treating hepatitis delta or a related viral infection in a mammalian patient in need of such treatment which comprises administering to said patient an anti-viral effective amount of a compound of claim 1.

30. A method of preventing restenosis in a mammalian patient in need of such treatment which comprises administering to said patient an amount of a compound of claim 1 which is effective for preventing restenosis.

31. A method of treating polycystic kidney disease in a mammalian patient in need of such treatment which comprises administering to said patient an amount of a compound of claim 1 which is effective for treating polycystic kidney disease..

32. A method of treating or preventing a disease selected from cancer, neurofibromin benign proliferative disorder, blindness related to retinal vascularization, infections from hepatitis delta and related viruses, restenosis and polycystic kidney disease, in a mammalian patient in need of such treatment which comprises administering to said patient an effective amount of a a compound of claim 1.