CA2249617A1 - 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

W O 97/36891 PCT~US97/05707 TITLE OF THE ~NVENTION
INHIBITORS OF FARNESYL-PROTElN TRANSFERASE

BACKGROUND OF THE INVENTION
S The Ras proteins (Ha-Ras, Ki4a-Ras, Ki4b-Ras and N-Ras) are part of a signaling pathway that links cell surface growth factor receptors to nuclear signals initiating cellular proliferation.
Biological and biochemical studies of Ras action indicate that Ras functions like a G-regulatory protein. Ln the inactive state, Ras is bound to GDP. Upon growth factor receptor activation Ras i,s induced to exchange GDP for GTP and undergoes a conformational change. The GTP-bound form of Ras propagates the growth stimulatory signal until the signal is termin~ted by the intrinsic GTPase activity of Ras, which returns the protein to its inactive GDP bound form (D.R. Lowy and D.M. Willumsen, Ann. Rev. Biochem. 62:851-~91 (1993)). Mutated ~as genes (Ha-ras, Ki4a-ras, Ki4b-ras and N-ras) are found in many human cancers, including colorectal carcinoma, exocrine pancreatic carcinoma, and myeloid leukemias. The protein products of these genes are defective in their GTPase activity and constitutively 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-terminus of Ra.s. The Ras C-terminus contains a sequence motif termed a "CAAX" or "Cys-Aaal-Aaa2-Xaa"
box (Cy.s is cysteine, Aaa is an aliphatic amino acid, the Xaa is any amino acid) (Willumsen et al., Natu~ e 310:5~3-5~6 ( 19~4)). Depending on the specific sequence, this motif serves as a signal sequence for the enzymes farnesyl-protein transferase or geranylgeranyl-protein transferase, which catalyze the alkylation of the cysteine residue of the CAAX motif with a Cls or C20 isoprenoid, re.spectively. (S. Clarke., Ann. Rev. Biochem.
61:355-3~6 (1992); W.R. Schafer and J. Rine, Ann. Rel~. Geneties 30:209-237 ( 1992)). The Ras protein is one of several proteins that are known to undergo post-translational farnesylation. Other farnesylated protein.s W O97136891 PCTrUS97/05707 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., J. Biol. Chem. 269, 141X2 (1994) have identified a peroxisome associated protein Pxf which is also farnesylated. James, et al., have also 5 suggested that there are farnesylated proteins of unknown structure and function in addition to those listed above.
Inhibition of farnesyl-protein transferase has been shown to block the growth of Ras-transformed cells in soft agar and to modify other aspects of their transformed phenotype. It has also been demon-10 strated that certain inhibitors of farnesyl-protein transferase selectively block the processing of the Ras oncoprotein intracellularly (N.E. Kohl et al., Scienc~e, 260:1934-1937 (1993) and G.L. James et al., Science, 260:1937-1942 (1993). Recently, it has been shown that an inhibitor of farnesyl-protein transferase blocks the growth of ras-dependent tumors in 15 nude mice (N.E. Kohl et al., Proc. Natl. Acad. Sci U.S.A., 91:9141-9145 (1994) and induces regression of m~mm~ry and salivary carcinomas in ras transgenic mice (N.E. Kohl et al., Nature Medicine, 1 :792-797 (1 995).
Indirect inhibition of farnesyl-protein transferase in l ivo 20 has been demonstrated with lovastatin (Merck & Co., Rahway, NJ) and compactin (Hancock et al., ibid; Casey et al., ibid; Schafer et al., Science 245:379 (1989)). These drugs inhibit HMG-CoA reductase, the rate limiting enzyme for the production of polyisoprenoids including farnesyl pyrophosphate. Farnesyl-protein transferase utilizes farnesyl 25 pyrophosphate to covalently modify the Cys thiol group of the Ras CAAX box with a farnesyl group (Reiss et al., Cell, 62:81-88 (1990);
Schaber et al., J. Biol. Chem., 265: 14701 -14704 (1990); Schafer et al., Science, 249: 1133- 1139 (1990); Manne et ~1., Proc . Natl. Acad. Sci USA.
87:7541 -7545 (1990)). Inhibition of farnesyl pyrophosphate biosynthesis 30 by inhibiting HMG-CoA reductase blocks Ras membrane localization in cultured cells. However, direct inhibition of farnesyl-protein transferase would be more specific and attended by fewer side effects than would occur with the required dose of a general inhibitor of isoprene biosynthesis.

WO 97/36891 PCT~US97/05707 Inhibitors of farnesyl-protein transferase (FPTase) have been described in two general classes. The fir~st are analogs of farnesyl diphosphate (FPP), while the second class of inhibitors is related to the protein substrates (e.g., Ras) for the enzyme. The peptide derived S inhibitors that have been described are generally cysteine containing molecules that are related to the CAAX motif that is the signal for protein prenylation. (Schaber et al., ibid; Reis.s et. al., ihid; Reiss et al., PNAS, 88:732-736 (1991)). Such inhibitors may inhibit protein prenylation while serving as alternate substrates for the farnesyl-protein transferase 10 enzyme, or may be purely competitive inhibitors (U.S. Patent 5,141,~51, University of Texas; N.E. Kohl et al ., Science, 260: 1934- 1937 (1993);
Graham, et al., J. Med. Chem., 37, 725 (1994)). In general, deletion of the thiol from a CAAX derivative has been shown to dramatically reduce the inhibitory potency of the compound. However, the thiol group 15 potentially places limitations on the therapeutic application of FPTase inhibitors with respect to pharmacokinetics, pharmacodynamics and toxicity. Therefore, a functional replacement for the thiol is desirable.
It has recently been reported that farnesyl-protein tran.sferase inhibitors are inhibitor.s of proliferation of vascular smooth muscle cell.s 20 and are therefore useful in the prevention and therapy of arteriosclero,sis and diabetic disturbance of blood vessels (JP ~7- 112930).
It has recently been disclosed that certain tricyclic compounds which optionally incorporate a piperidine moiety are inhibitors of FPTase (WO 95/10514, WO 95/10515 and WO 95/10516).
25 Imidazole-containing inhibitors of farnesyl protein transferase have also been disclosed (WO 95/09001 and EP 0 675 112 Al ).
It is, therefore, an object of this invention to develop peptidomimetic compounds that do not have a thiol moiety, and that will inhibit farnesyl-protein transferase and thus, the post-translational 30 farnesylation of proteins. It is a further object of this invention to develop chemotherapeutic compo,sitions containing the compounds of this invention and methods for producing the compounds of this invention.

SUMMARY OF THE INVENTION
The present invention comprises analogs of the CA1A2X
motif of the protein Ras that is modified by farnesylation in vivo. These CA 1 A2X analogs inhibit the farnesylprotein transferase. Furthermore, 5 these CA 1 A2X analogs differ from those previously described as inhibitors of farnesyl-protein transferase in that they do not have a thiol moiety. The lack of the thiol offers unique advantages in terms of improved pharmacokinetic behavior in ~nim~ , prevention of thiol-dependent chemical reactions, such as rapid autoxidation and disulfide 10 formation with endogenous thiols, and reduced systemic toxicity. The compounds of the instant invention also incorporate a cyclic amine moiety in the A2 position of the motif. The compounds of the instant invention also do not contain a carboxylic acid, and therefore do not require a prodrug ester for improved cell permeability. Further contained 15 in this invention are chemotherapeutic compositions containing these farnesyl transferase inhibitors and methods for their production.

The compounds of this invention are illustrated by the formulae:

(R8) /~9\ Z

V ~ A1(CR1az)nA2(cR1 2)n -\w/ - (CRlb2)p~N~N

DETAILED DESCRIPTION OF THE INVENTION
The compounds of this invention inhibit the farnesyl-protein 25 transfera.se. In a first embodiment of this invention, the farnesyl-protein transferase inhibitors are illustrated by the formula 1:

(R8)r /R9\ Z

V - A1 (C R1 a2)nA2(C R l a2) n ~W/ - (C R l b2)~ N~N~RRs5b R4b R4a wherein:
Rla and R1b are independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, alkenyl, alkynyl, R10O-, Rl 1S(o)m, R10C(O)NRl0-~ CN, NO2, (R 1 0)2N-C(NR 10) , R 1 ~C(O)-, CON(R 1~)2-, N3, -N(R10)2, or Rl lOC(O)NR10-c) Cl-C6 alkyl unsubstituted or substituted by aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl, R100-, Rl 1S(O)m-, R10C(o)NR10-, CN, (R10)2N-C(NR10)-, R10C(o)-, CON(R10)2-, N3, -N(R10)2, or Rl loC(O)-NR10-;
R2 and R3 are independently selected from:
a) a side chain of a naturally occurring amino acid, b) an oxidized form of a side chain of a naturally occurring amino acid which is:
i) methionine sulfoxide, or ii) methionine sulfone, and c) substituted or unsubstituted C 1 -C20 alkyl, C2-C20 alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, N(R 1~)2, NO2, R 10o-, R 1 1 S(O)m ~ R 1 0C(o)NR 10, CN, (R 10)2N-C(NR 10), R10c(o)-~ CON(R10)2-, N3, -N(R10)2, Rl lOC(O)NR10- and Cl-C20 alkyl, and d) Cl-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-CIo cycloalkyl; or S R2 and R3 are combined to form - (CH2)S -; or R2 or R3 are combined with R6 to form a ring such that R6 ~
~; N ~ is ~ H2)t ;
R2 R3 R7a~R7b R4a, R4b, R7a and R7b are independently selected from:
a) hydrogen, b) C1 -C6 alkyl unsubstituted or substituted by alkenyl, R 10o-, Rl lS(O)m-, RlOC(O)NR10-, CN, N3, (R10)2N-C(NlR10)-, R1OC(O)-, CON(R10)2-, -N(R10)2, or Rl lOC(O)NR10-, c) aryl, heterocycle, cycloalkyl, alkenyl, RlOO, Rl lS(o)m, Rloc(o)NRlo-~ CN, NO2, 1 0)2N C(NR 10), R 1 ~C(O)-, CON(R 1~)2-, N3, -N(R10)2, or Rl lOC(O)NR10-, and d) Cl-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-Clo cycloalkyl;

R5a and R5b are independently selected from:
a) hydrogen, b) substituted or unsubstituted C 1 -C20 alkyl, C2-C20 alkenyl, C3-Clo cycloalkyl, aryl or heterocycle group, wherein the substituent is selected from F, Cl, Br, (R 1 0)2NC(O)-, NO2, R l Oo ~ R l l S(o)m R 1 OC(O)NR 10, CN, (R 1 0)2N-C(NR 1 0) , R 1 ~C(O)-, CON(R 1 0)2-? N3, -N(R 1 0)2, R l l OC(O)NR 1 0- and Cl-C20 alkyl, d) Cl-C6 alkyl sub,stituted with an un~substituted or substituted group selected from aryl, heterocycle and C3-CIo cycloalkyl; or RSa and RSb are combined to form - (CH2)S - wherein one of the carbon atoms is optionally replaced by a moiety selected from:
O, S(O)m, -NC(O)-, and -N(COR10)-;
R6 is independently selected from hydrogen or Cl-C6 alkyl;

R~ is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R10O-, Rl lS(O)m-, R 1 0C(O)NR 10, CN, NO2, R 1 02N-C(NR 10) , R l ~C(O)-, CON(R10)2-, N3, -N(R10)2, or Rl 1OC(O)NR10-, and c) Cl-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R10O-, Rl lS(O)m-, R10C(O)NH-, CN, H2N-C(NH)-, R10C(o)-, CON(R10)2-, N3, -N(R10)2, or CON(R 1 0)2NH-;

25 R9 is selected from:
a) hydrogen, b) alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R10O-, Rl lS(o)m, R10C(O)NR10-~ CN, NO2, (R 1 0)2N-C-(NR 10), R 1 ~C(O)-, CON(R 1~)2-, N3, -N(R 1 0)2, or R l l OC(O)NR 1 0-, and c) Cl-C6 alkyl unsubstituted or ,substituted by perfluoroalkyl, F, Cl, Br, R10O-, RllS(O)m-, R10C(o)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3, -N(R 1~)2, or R 1 1 OC(O)NR 10;

W O97/36891 PCTrUS97/05707 _ " _ R10 i,s independently selected from hydrogen, C1-C6 alkyl, benzyl and aryl;

5 Rl 1 is independently selected from Cl-C6 alkyl and aryl;

A 1 and A2 are independently selected from: a bond, -CH=CH-, -C-C-, -C(0)-, -C(O)NR 10, -NR 1 ~C(0)-, 0, -N(R 10), -S(0)2N(R10)-, -N(RlO)S(0)2-~ or S(O)m;
Q i,s a substituted or uulsubstituted nitrogen-cont~ining C4-Cg mono orbicyclic ring system, wherein the non-nitrogen containing ring may be a Cs-C7 saturated ring;

15 V is selected from:
a) hydrogen, b) heterocycle, c) aryl, d) C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from 0, S, and N, and e) C2-C20 alkenyl, provided that V is not hydrogen if A 1 is S(0)m and V is not hydrogen if Al is a bond, n is 0 and A2 is S(O)m;

25 W is a heterocycle;

X, Y and Z are independently H2 or 0;

m is 0, 1 or 2;
nis 0,1,2,30r4;
pis 0, 1,2,30r4;
r is 0 to 5, provided that r is 0 when V is hydrogen;
s is 4 or S;
tis 3,40rS; and W O97/36891 PCT~US97/05707 uis Oor l;
or the pharmaceutically acceptable salts, crystal forms, hydrates and isomers thereof.
In a more preferred embodiment of this invention, the Ras S farnesyl transferase inhibitors are illustrated by the Fo~nula I:

(R8)r /R9\ Z

V - A1(Cl~l ~)nA~(CR1a2)n -\w/ - (CR1b2)~

wherem:
Rla is independently selected from: hydrogen or Cl-C6 alkyl;

Rlb is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, RlOo, -N(R 1~)2 or alkenyl, c) Cl-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, R100-, or -N(R10)2;

R2 and R3 are independently selected from:
a) a side chain of a naturally occurring amino acid, b) an oxidized form of a side chain of a naturally occurring amino acid which is:
i) methionine sulfoxide, or ii) methionine sulfone, c) substituted or unsubstituted Cl-Clo alkyl, C2-Clo alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, N02, R 1 00-, R l l S(O)m-, R l OC(O)NR 10-, CN, WO 97/36891 PCTrUS97/05707 (R 1 0)2N-C(NR 10) , R 1 ~C(O)-, CON(R 1~)2-, N3, -N(R10)2, Rl IOC(o)NRlo- and Cl-C20 alkyl, and d) Cl-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C1 o cycloalkyl; or R2 and R3 are combined to form - (CH2),s -; or R2 or R3 are combined with R6 to form a ring such that R6 R7a~R7b;

R4a and R7a are independently selected from:
a) hydrogen, l~S b) Cl-C6 alkyl unsubstituted or substituted by alkenyl, R100-, R1 1S(O)m-, R10C(o)NR10-, CN, N3, (R10)2N-C(NR10)-, R 1 0c(O)-, CON(R 1 0)2-, -N(R 1~)2, or R l 1 OC(O)NR 1 0-, c) aryl, heterocycle, cycloalkyl, alkenyl, R10O-, R 1 1 S(O)m-~ R 1 0C(O)NR 10, CN, NO2, (R10)2N C(NR10), R10C(o), CON(R10)2-, N3, -N(R 1~)2, or R 1 1 OC(O)NR 10, and d) Cl -C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-Clo cycloalkyl;
R4b and R7b are hydrogen;

RSa is selected from:
a) substituted or unsubstituted C 1 -C 10 alkyl, C2-C1 0 alkenyl, 3~ C3-Clo cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, N02, R100-, Rl lS(O)m-, RlOC(O)NR10-, W O 97/36891 PCTrUS97/05707 (R1o)2Nc(o)-~ CN, (R1o)2N-c(NRlo)-~ R10C(o)-, CON(R10)2-, N3, -N(R10)2, Rl lOC(O)NR10- and Cl-C20 alkyl, and b) C 1 -C6 alkyl .substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-Clo cycloalkyl;

R5b is .selected from:
a) hydrogen, and b) Cl-C3 alkyl;
R6 is independently selected from hydrogen or C 1 -C6 alkyl;

R~ is independently selected from:
a) hydrogen, b) Cl-C6 alkyl, C2-c6 alkenyl, C2-c6 alkynyl, Cl-c6 perfluoroalkyl, F, Cl, R100-, RlOC(O)NR10-, CN, N02, (R10)2N-C(NR10)-, R10C(o)-, CON(R10)2-, -N(R10)2, or R 1 1 OC(O)NR 10, and c) C l -C6 alkyl substituted by C l -C6 perfluoroalkyl, R 1 0O-, RlOC(o)NRlo-~ (R10)2N-C(NR10)-, RlOC(O)-.
CON(R 1~)2-, -N(R 1~)2, or R 1 1 OC(O)NR 10;

R9 is selected from:
a) hydrogen, b) C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl, R10O-, R1 lS(O)m-, R10C(O)NRl0-~ CN, NO2, (R 1 0)2N-C(NR 10), R I ~C(O)-, CON(R 1~)2-, -N(R 1~)2, or R l l OC(O)NR 1 0-, and c) Cl -C6 alkyl unsubstituted or substituted by Cl -C6 perfluoroalkyl, F, Cl, R 1 0O-, R 1 I S(O)m-, R 1 0C(O)NR 1 0-, CN, (R10)2N-C(NR10)-, R10C(o)-, CON(R10)2-, -N(R10)2, or Rl lOC(O)NR10-;

W O97/36891 PCTrUS97/05707 R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl and aryl;
R11 is independently selected from Cl-C6 alkyl and aryl;
Q is selected from:

N~ and -~- N~

~0 AI and A2areindependentlyselectedfrom: abond,-CH=CH-,-C-C-, -C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;

V is selected from:
a) hydrogen, b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, c) aryl, d) Cl-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and e) C2-C20 alkenyl, and provided that V is not hydrogen if A 1 is S(O)m and V is not hydrogen if A 1 is a bond, n is 0 and A2 is S(O)m;

25 W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, 4uinolinyl, or isoquinolinyl;

- X, Y and Z are independently H2 or O;
mis 0,1 or2;

W O 97/36891 PCTrUS97/05707 nis 0, 1,2,30r4;
pis 0, 1,2,30r4;
r is O to 5, provided that r is O when V is hydrogen;
tis 3,40r5;and uis Oorl;
or the pharmaceutically acceptable salts, hydrates, crystal forms and isomers thereof.
In an even more preferred embodiment of this invention, the Ras farnesyl transferase inhibitors are illustrated by the Formula II:

V Al(CRla2)nA2(CR1a2),.-(W)- (CR1b2)p~N~N~' R4a wherein:
Rla is independently selected from: hydrogen or Cl-C6 alkyl;

Rlb is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, R 10o-, -N(R 1 ~)2 or alkenyl, c) Cl-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, R100-, or -N(R10)2;

R2 or R3 are combined with R6 to form a ring such that ~;N~, iS ~H2~t;
R2 R3 R7a R7b R4a is independently selected from:

WO 97/36891 PCT~US97/05707 a) hydrogen, b) C 1 -C6 alkyl unsubstituted or sub.stituted by alkenyl, R 1 0O-, Rl lS(O)m-, R10C(o)NRl0-, CN, N3, (R10)2N-C(NR10)-, R 10c(O)-, CON(R 1~)2-, -N(R10)2, or R 1 lOC(O)NR 10, c) aryl, heterocycle, cycloalkyl, alkenyl, R10O-, Rl lS(O)m-, R10C(O)NR10-, CN, NO2, (R 10)2N C(NR10), R 10c(O)-, CON(Rl0)2-~ N3, -N(R 1 0)2, or R1 1 OC(O)NR 1 0-, and d) Cl-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-C1o cycloalkyl;

R4b and R7b are hydrogen;

RSa is selected from:
a) substituted or unsubstituted Cl-C1o alkyl, C2-Clo alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, NO2, R 1 0O, R 1 1 S(O)m-, R 1 0C(O)NR 1 0-, (R10)2Nc(o)-~ CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R 1~)2-, N3, -N(R 1~)2, R 1 1 OC(O)NR 10 and Cl-C20 alkyl, and b) C 1 -C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-Clo cycloalkyl;

RSb i~s selected from:
a) hydrogen, and b) C1-C3 alkyl;
R6 is independently selected from hydrogen or C1-C6 alkyl;

- R8 is independently selected from:
a) hydrogen, WO97/36891 PCT~US9710~707 b) Cl-C6 alkyl, C2-c6 alkenyl, C2-c6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl, R10O-, R10C(o)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(o)-, CON(R10)2-, -N(R10)2, or ~ R 1 1 OC(O)NR 10, and c) Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl, R10O-, RlOC(o)NRlo-~ (R10)2N-C(NR10)-, RlOC(O)-, CON(R10)2-, -N(R10)2, or Rl lOC(O)NR10-;

R9 is .selected from:
a) hydrogen, b) C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl, R10O-, Rl lS(O)m-, R10C(O)NRl0-~ CN, NO2, (R10)2N-C(NR10)-, R10C(o)-, CON(R10)2-, -N(R10)2, or R 1 1 OC(O)NR 10, and c) Cl-C6 alkyl unsubstituted or substituted by Cl-C6 perfluoroalkyl,~F, Cl, R 1 0O-, R 1 1 S(O)m-~ R 1 0C(O)NR 10, CN, (R10)2N-C(NR10)-, R10C(o)-, CON(R10)2-, -N(R10)2, or Rl lOC(O)NR10-;

20 R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl and aryl;
R l 1 is independently selected from Cl-C6 alkyl and aryl;
25 Q is selected from:

~r N~ and -~- N~

A l and A2 are independently selected from: a bond, -CH=CH-~ -C_C-, -C(O)-, -C(O)NR 10, o, -N(R 10) , or S(O)m;

W O 97/36891 PCT~US97/05707 V is selected from:
a) hydrogen, b) heterocycle ~selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, 4uinolinyl, isoquinolinyl, and thienyl, c) aryl, d) C1-C20 alkyl wherein from O to 4 carbon atoms are replaced with a heteroatom selected from 0, S, and N, and e) C2-C20 alkenyl, and 10 provided that V is not hydrogen if A 1 is S(O)m and V is not hydrogen if Al is a bond, n i,s O and A2 is S(O)m;

W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or 1 S isoquinolinyl;

X, Y and Z are independently H2 or 0;

m is 0, 1 or 2;
nis 0,1,2,30r4;
pis 0, 1,2,30r4;
r is O to 5, provided that r is O when V is hydrogen;
tis 3,40r5;and uis Oorl;
25 or the pharmaceutically acceptable salts, hydrates, crystal forms or isomers thereof.
In yet a more preferred embodiment of this invention, the Ras farnesyl transferase inhibitors are illustrated by the Formula rv:

CA 022496l7 l998-09-22 W 097/36891 PCT~US97tO5707 V A1(CR1a2)nA2(C~1a2)n-(W)- (CR1b2)p~l~N~ R

lV O R4b R a wherein:
5 Rla is independently selected from: hydrogen or C1-C6 alkyl;

R 1 b is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, R100-, -N(R 1~)2 or alkenyl, c) C1-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, R10O-, or-N(R10)2;

R4a and R7a are independently selected from:
a) hydrogen, b) C1 -C6 alkyl unsubstituted or substituted by alkenyl, R 10o-, Rl lS(O)m-, R10C(O)NRl0-~ CN, N3, (R10)2N-C(NR10)-, R 1 ~C(O)-, CON(R 1~)2-, -N(R 1~)2, or R I 1 OC(O)NR 10 , c) aryl, heterocycle, cycloalkyl, alkenyl, R10O-, R 1 1S(O)m-, R10C(o)NR10-, CN, NO2, (R10)2N-C(NR 10), R 10c(O)-, CON(R 1~)2-, N3, -N(R10)2, or R1 1OC(O)NR10-, and d) C 1 -C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-Clo cycloalkyl;
- R4b is hydrogen;
RSa is ~selected from:
a) substituted or unsub~stituted Cl-C1o alkyl, C2-Clo alkenyl, C3-Cl o cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, NO2, R l Oo, R I 1 S(O)m-~ R I OC~O)NR 10, (R 1 0)2NC(o)-, CN, (R 1 0)2N-C(NR 10) , R 1 ~C(O)-, CON(R10)2-, N3, -N(R10)~, Rl lOC(O)NR10- and Cl-C20 alkyl, and b) C l-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-Clo cycloalkyl;
~0 Rsb is selected from:
a) hydrogen, and b) C 1 -C3 alkyl;

R8 is independently selected from:
a) hydrogen, b) Cl-C6 alkyl, C2-c6 alkenyl, C2-C6 alkynyl, Cl-c6 perfluoroalkyl, F, Cl, R100-, R10C(O)NR10-,CN,NO2, (Rl0)2N-c(NRlo)~Rloc(o)-~coN(Rlo)2-~-N(Rlo)2~ or RllOC(O)NR10-, and c) C1 -C6 alkyl substituted by Cl -C6 perfluoroalkyl, Rl Oo, Rl0c(o)NRlo-~(Rlo)2N-c(NRlo)-~Rloc(o) CON(R10)2-, -N(R10)2, or Rl lOC(O)NR10-;

R9is selected from:
a) hydrogen, b) C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 pe~luoroalkyl, F,CI,R100-,RllS(O)m-,R10C(O)NR10-,CN,NO2, (Rl0)2N-c(NRlo)-~Rloc(o)-~coN(Rlo)2-~-N(Rlo)2~ or RllOC(O)NR10-, and c) C 1 -C6 alkyl unsub~tituted or substituted by C 1 -C6 perfluoroalkyl, F, Cl, R1OO-, RllS(O)m-,Rl0c(o)NR
CN,(Rl0)2N-c(NRlo)-~Rloc(o)-~coN(Rlo)2 -N(R10)2, or Rl lOC(O)NR10-;

W O97/36891 PCT~US97/0~707 R10 i.s independently selected from hydrogen, Cl-C6 alkyl, benzyl and aryl;
~ 11 is independently selected from Cl -C6 alkyl and aryl;

Q is selected from:

N~ and -~- N _ ~

~0 Al and A2 areindependentlyselectedfrom: abond, -CH=CH-, -C_C-, -C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;

V is selected from:
a) hydrogen, lS b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, c) aryl, d) C l -C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and e) C2-C20 alkenyl, and provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if Al is a bond, n is 0 and A2 is S(O)m;

2;S W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or isoquinolinyl;

- Z is independently H2 or O;
m is 0, 1 or 2;

W O 97/36891 PCT~US97/05707 nis 0,1,2,30r4;
pi~ 0, 1,2,30r4;
r is O to 5, provided that r is O when V is hydrogen;
tis 3,40r5; and uis Oor l;
or the pharmaceutically acceptable salts, hydrates, crystal forms, or isomers thereof.
In the most preferred embodiment of this invention. the Ras farnesyl transferase inhibitors are illustrated by the Formula V:

V A (CR 2)nA (CR1a2)n -(W)- (CR1b2)p~fN~--N/~ 5b IV O
wherein:
15 Rla is independently selected from: hydrogen or Cl-C6 alkyl;

R1b is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, R100-, -N(R10)2 or alkenyl, c) Cl-C6 alkyl un~substituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, R100-, or -N(R10)2;

R4a and R7a are independently .selected from:
a) hydrogen, b) C l -C6 alkyl unsubstituted or substituted by alkenyl, R l Oo, Rl lS(O)m-, RlOC(O)NRlO-, CN, N3, (R10)2N-C(NR10)-, R 1 ~C(O)-, CON(R 1~)2-, -N(R 1~)2, or R 1 1 OC(O)NR 10, c) aryl, heterocycle, cycloalkyl, alkenyl, R100-, R 1 l S(O)m, R l OC(O)NR 1 0-, CN, N02, (R10)2N-C(NR10)-, RlOC(O)-, CON(R10)2-, N3, -N(R 1~)2, or Rl lOC(O)NR10-, and W 097/36891 PCTrUS97/05707 d) C I -C6 alkyNsubstituted with an un,substituted or substituted group ,selected from aryl, heterocyclic and C3-Clo cycloalkyl;

5 R5a is selected from:
a) substituted or unsubstituted Cl-C1o alkyl, C2-Clo alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, NO2, R10O-, Rl lS(O)m-, R10C(o)NR10-, (R 1 0)2NC(O)-, CN, (R 1 0)2N-C(NR 10) , R 1 ~C(O)-, CON(R 1~)2-, N3, -N(R 1~)2, R 1 1 OC(O)NR 10 and C1-c2o alkyl, and b) Cl -C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C1o cycloalkyl;

R5b is selected from:
a) hydrogen, and b) C 1 -C3 alkyl;
R~ is independently selected from:
a) hydrogen, b) C 1 -C6 alkyl, C2-c6 alkenyl, C2-c6 alkynyl, C 1 -C6 perfluoroalkyl, F, Cl, R10O-, R10C(O)NRl0-~ CN, NO2, (R 1 0)2N-C(NR 10)-, R 1 ~C(O)-, CON(R 1~)2-, -N(R 1~)2, or R 1 1 OC(O)NR 10-, and c) C I -C6 alkyl substituted by C 1 -C6 perfluoroalkyl, R 1 0O-, RlOC(O)NR10-, (R10)2N-C(NR10)-, RlOC(O)-, CON(R 1~)2-, -N(R 1~)2, or R 1 1 OC(O)NR 10;
R9 is selected from:
a) hydrogen, b) C2-C6 alkenyl, C2-C6 alkynyl, C 1 -C6 perfluoroalkyl, W O97/36891 PCTrUS97/05707 F~ Cl, R100-, R~ lS(O)m-, RlOC(O)NR10-, CN, N02, (R 1 0)2N-C(NR 10), R I ~C(O)-, CON(R 1~)2-, -N(R 1~)2, or R 1 1 OC(O)NR 10, and c) Cl-C6 alkyl unsubstituted or substituted by Cl-C6 S perfluoroalkyl, F, Cl, R 1 00-, R 1 l S(O)m-, R l OC(O)NR 10, CN, (R 10)2N-C(NR 10), R 10C(o)-, CON(R10)2-, -N(R10)2, or Rl lOC(O)NR10-;

R10 is independently selected from hydrogen, Cl-C6 alkyl, benzyl and aryl;
R 1 1 is independently selected from Cl -C6 alkyl and aryl;

A 1 and A2 are independently selected from: a bond, -CH=CH-, -C_C-, -C(O)-, -C(O)NR 10, O, -N(R10)-, or S(O)m;

V is selected from:
a) hydrogen, b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, c) aryl, d) Cl-C20 alkyl wherein from O to 4 carbon atoms are replaced with a heteroatom selected from 0, S, and N, and e) C2-C20alkenyl~ and provided that V is not hydrogen if A 1 is S(O)m and V is not hydrogen if A1 is a bond, n is O and A2 i.s S(O)m;

W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, 30 thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or isoquinolinyl;

m is 0, 1 or 2;
nis 0, 1,2,30r4;
pis 0,1,2,30r4;

W O97136891 PCTrUS97/05707 r is O to 5, provided that r is O when V is hydrogen:
tis 3,40r5; and uis Oor l;
or the pharmaceutically acceptable .salt.s, hydrates, crystal form~s or 5 isomers thereof.

Examples of the compounds of this invention are as follows:
N-[ 1-[1 -(4-Cyanobenzyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-(3-chlorophenylmethyl)-amide N- [ 1 - [ 1 -(4-Cyanobenzyl)- 1 H-imidazol-S -ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-methyl-N-(3-chlorophenylmethyl)-amide N-Ll-l 1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]-3(S)-ethylpyrrolidin-15 2(S)-ylmethyl]-3(S)-ethyl-proline N-(3-chlorophenylmethyl)-amide N-[l-[l -(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-methyl-N-(3-chlorophenylmethyl)-amide N-[1-(3-l IH-Imidazol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[ 1-(3-[1 H-Imidazol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-25 ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[ 1-( I -( I -Farnesyl)- 1 H-imidazol-5-ylacetyl)-pyrrolidin-2(S)-ylrnethyl] -3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide 30 N-[ 1-(1-( I -Geranyl)- I H-imidazol-5-ylacetyl)-pyrrolidin-2(S)-ylmethyl] -3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide - N-[1-[1-(4-Methoxybenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-~(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide W O 97/36891 PCT~US97/05707 N-[ 1-[1 -(4-Methoxybenzyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[ 1-~ I -(2-Naphthylmethyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl3-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide N-[ 1-[1 -(2-Naphthylmethyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyll-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide 10 or the pharmaceutically acceptable salts thereof.

Specific examples of compound,s of the invention are:
N-[ 1-(3-[1 H-Imidazol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide Cl 0 ~ 9--N~
N--N~

N-[l -[1-(4-Cyanobenzyl)-lH-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-(3-chlorophenylmethyl)-amide NC

~~

W O 97t36891 PCTAUS97/05707 N-[ I -L 1 -(4-Cyanobenzyl)- 1 H-imidazol-~-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-methyl-N-(3-chlorophenylmethyl)-amide - NC

~r~

or the pharmaceutically acceptable salts thereof.
In the present invention, the amino acids which are disclosed are identified both by conventional 3 letter and single letter abbreviations as indicated below:
Alanine Ala A
Arginine Arg R
Asparagine Asn N
Aspartic acid Asp D
Asparagine or Asparticacid Asx B
Cysteine Cys C
Glutamine Gln Q
Glutamic acid Glu E
Glutamine or Glutamic acid Glx Z
Glycine Gly G
Histidine His H
Isoleucine Ile ~5 Leucine Leu L
Lysine Lys K
Methionine Met M

W O97/36891 PCT~US97/05707 Phenylalanine Phe F
Proline Pro P
Serine Ser S
Threonine Thr T
Tryptophan Trp W
Tyrosine Tyr Y
Valine Val V

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.
As used herein, "alkyl" is intended to include both branched 15 and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms.
As used herein, "cycloalkyl" is intended to include non-aromatic cyclic hydrocarbon groups having the specified number of carbon atoms. Examples of cycloalkyl groups include cyclopropyl, 20 cyclobutyl, cyclopentyl, cyclohexyl and the like.
"Alkenyl" groups include those groups having the specified number of carbon atoms and having one or several double bonds.
Examples of alkenyl groups include vinyl, allyl, isopropenyl, pentenyl, hexenyl, heptenyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclo-25 hexenyl, l-propenyl, 2-butenyl, 2-methyl-2-butenyl, isoprenyl, farnesyl, geranyl, geranylgeranyl and the like.
As used herein, "aryl" is intended to include any stable monocyclic, bicyclic or tricyclic carbon ring(s) of up to 7 members in each ring, wherein at least one ring is aromatic. Examples of aryl groups 30 include phenyl, naphthyl, anthracenyl, biphenyl, tetrahydronaphthyl, indanyl, phenanthrenyl and the like.
The term heterocycle or heterocyclic, as used herein, represents a stable 5- to 7-membered monocyclic or stable ~- to 11-membered bicyclic or stable 1 1-15 membered tricyclic heterocycle ring W O 97/36891 PCTrUS97/05707 which is either saturated or unsaturated, and which consists of carbon atom~s and from one to four heteroatoms selected from the group consisting of N, O, and S, and including any bicyclic group in which any of the above-defined heterocyclic rings is fu~sed to a benzene ring.
5 The heterocyclic ring may be attached at any heteroatom or carbon atom which results in the creation of a stable structure. Examples of such heterocyclic elements include, but are not limited to, azepinyl, benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl, benzothiazolyl, benzothienyl, 10 benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydro-benzothienyl, dihydrobenzothiopyranyl, dihydrobenzothio-pyranyl sulfone, furyl, imidazolidinyl, imidazolinyl, irnidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, isothiazolidinyl, morpholinyl, naphthyridinyl, oxadiazolyl, 15 2-oxoazepinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, piperidyl, piperazinyl, pyridyl, pyridyl N-oxide, pyridonyl, pyrazinyl, pyrazolidinyl, pyrazolyl, pyrimidinyl, pyrrolidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinolinyl N-oxide, quinoxalinyl, tetrahydrofuryl, tetrahydro-isoquinolinyl, tetrahydro-quinolinyl, thiamorpholinyl, thiamorpholinyl 20 sulfoxide, thiazolyl, thiazolinyl, thienofuryl, thienothienyl, and thienyl.
As used herein, the terms "substituted aryl", "substituted heterocycle" and "substituted cycloalkyl" are intended to include the cyclic group which is .substituted with 1 or 2 substitutents selected from the group which includes but is not limited to F, Cl, Br, CF3, NH2 25 N(CI-C6 alkyl)2, NO2, CN, (Cl-C6 alkyl)O-, -OH, (Cl-C6 alkyl) S(O)m-, (C1-C6 alkyl)C(O)NH-, H2N-C(NH)-, (Cl-C6 alkyl)C(O)-, (Cl-C6 alkyl)OC(O)-, N3,(CI-C6 alkyl)OC(O)NH- and Cl-C20 alkyl.
The following structure:
,N~
((~H2)t J

- 2~ -represents a cyclic amine moiety having S or 6 members in the ring, such a cyclic amine which may be optionally fused to a phenyl or cyclohexyl ring. Examples of such a cyclic amine moiety include, but are not limited to, the following specific structures:

5 (~ ~ ~
It is also understood that substitution on the cyclic amine moiety by R~a and R8b may be on different carbon atoms or on the same carbon atom.
When R3 and R4 are combined to forrn - (CH2)S -, cyclic moieties are formed. Examples of such cyclic moieties include, but are 10 not limited to:

As used herein, the phrase "nitrogen containing C4-C9 mono or bicyclic ring system wherein the non-nitrogen containing ring may be a C5-C7 saturated ring" which defines moiety "Q" of the instant 15 invention includes but is not limited to the following ring sy.stems:

W 097136891 PCTrUS9~/05707 ~5- N~ N~

~_N~ -~--N~, The pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds S of this invention as formed, e.g., from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like: and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, 10 malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenyl-acetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, i~sethionic, trifluoroacetic and the like.
It is intended that the definition of any substituent or variable l~S (e.g., R10, Z, n, etc.) at a particular location in a molecule be independent of its definitions elsewhere in that molecule. Thus, -N(R 1~)2 represents -NHH, -NHCH3, -NHC2Hs, etc. It is understood that substituents and substitution patterns on the compounds of the instant invention can be selected by one of ordinary skill in the art to provide compounds that are 20 chemically stable and that can be readily synthesized by techniques known in the art a~s well as those methodl~i set forth below.

W O 97136891 PCTrUS97/05707 Preferably, Rla and Rlb are independently selected from:
hydrogen, -N(R~)2, R8C(O)NR8- or Cl-C6 alkyl unsubstituted or substi~ted by -N(R~)2, R~sO- or R8C(O)NRg-.
Preferably, R2 is the sidechain of glycine (hydrogen).
Preferably, R3 is selected from:
a) a side chain of a naturally occurring amino acid, b) substituted or unsubstitllted Cl -C20 alkyl, wherein the substituent is selected from F, Cl, Br, N(R 1 0)2, NO2, R 1 0O-, R 1 1 S (~)m-, R l 0C(o)NR 10, CN, (R 1 0~2N-C(NR 10) , R 1 ~C(O)-, CON(R 1~)2-, N3, -N(R10)2, R1 1OC(O)NR10- and C1-C20 alkyl, and c) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C1o cycloalkyl; or R3 is combined with R6 to form pyrrolidinyl ring.
Preferably, R4a, R4b, R7a and R7b are independently selected from: hydrogen, C1-C6 alkyl, aryl and benzyl.
Preferably, R5a and R5b are independently selected from:
hydrogen, unsubstituted or substituted C1-C6 alkyl, aryl, or Cl-3 alkyl substituted with an unsubstituted or substituted group selected from aryl or heterocycle .
Preferably, R6 is: hydrogen or is combined with R3 to form pyrrolidinyl ring.
Preferably, R~ is selected from: hydrogen, perfluoroalkyl, F, Cl, Br, R10O-, Rl 1S(O)m-, CN, NO2, R102N-C(NR10)-, Rl0C(o)-, CON(R10)2-, N3, -N(R10)2, or RllOC(O)NR10- and C1-C6 alkyl.
Preferably, R9 is hydrogen.
Preferably, R 10 is selected from H, C 1 -C6 alkyl and benzyl.
Preferably, R 12 is selected from C1 -C6 alkyl and benzyl.
Preferably, Al and A2 are independently selected from:
a bond, -C(O)NR 10, -NR l ~C(O)-, O, -N(R 10) , -S(O)2N(R 10) and -N(R I ~)S(O)2-.
Preferably, Q is a pyrrolidinyl ring.

W O97/36891 PCT~US97/05707 Preferably, V i.s selected from hydrogen, heterocycle and aryl.
Preferably, n, p and r are independently 0, 1, or 2.
Preferably t is 3.
The pharmaceutically acceptable salts of the compounds of this invention can be synthe.sized from the compounds of this invention which contain a basic moiety conventional chemical methods. Generally, the salts are prepared by ion exchange chromatography or by reacting the free base with stoichiometric amounts or with an excess of the desired 10 salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents.
The compounds of the invention can be synthesized from their constituent amino acids by conventional peptide synthesis techniques, and the additional methods described below. Standard 15 methods of peptide synthesis are di~sclosed, for example, in the following works: Schroeder et al., "The Peptides", Vol. I, Academic Press 1965, or Bodanszky et al., "Peptide Synthesi~", Interscience Publishers, 1966, or McOmie (ed.) "Protective Groups in O~ganic Chemistry", Plenum Press, 1973, or Barany et al., "The Peptides: Analysis, Synthesis, Biology" 2, 20 Chapter 1, Academic Press, 1980, or Stewart et al., "Solid Phase Peptide Synthesis", Second Edition, Pierce ChemicaJ Company, 1984. The teachings of these works are hereby incorporated by reference.
Abbreviations used in the description of the chemistry and in the Examples that follow are:
Ac2O Acetic anhydride;
Boc t-Butoxycarbonyl;
DBU 1,~-diazabicyclo[5.4.0]undec-7-ene;
DMAP 4-Dimethylaminopyridine;
DME 1,2-Dimethoxyethane;
DMF Dimethylformamide;
EDC 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide-hydrochloride;
HOBT I -Hydroxybenzotriazole hydrate;

Et3N Triethylamine;
EtOAc Ethyl acetate;
FAB Fast atom bombardment;
HOOBT 3-Hydroxy- 1 ,2,2-benzotriazin-4(3~)-one;
5 HPLC High-performance li4uid chromatography;
MCPBA m-Chloroperoxybenzoic acid;
MsCI Methane~sulfonyl chloride;
NaHMDS Sodium bis(trimethylsilyl)amide;
Py Pyridine;
TFA Trifluoroacetic acid;
THF Tetrahydrofuran.

Compounds of this invention are prepared by employing the reactions shown in the following Reaction Schemes A-J, in addition to other standard manipulations such as ester hydrolysis, cleavage of protecting groups, etc., as may be known in the literature or exemplified in the experimental procedures. Some key bond-forming and peptide modifying reactions are:

Reaction A Amide bond formation and protecting group cleavage using standard solution or solid phase methodologies.

Reaction B Preparation of a reduced peptide subunit by reductive alkylation of an amine by an aldehyde using sodium cyanoborohydride or other reducing agents.

Reaction C Deprotection of the reduced peptide subunit Reaction D Amide bond formation and protecting group cleavage using standard solution or solid phase methodologies.
~ Reaction E Preparation of a reduced subunit by borane reduction of the amide moiety.

W O97136891 PCT~US97/05707 Reaction Schemes A-E illustrate bond-forming and peptide modifying reactions incorporating acyclic peptide units. It is well under-stood that such reaction,s are equally useful when the - NHC(RA)- moiety of the reagents and compounds illustrated is replaced with the following 5 moiety:

(C~ H2)t ~R7b R7a These reactions may be employed in a linear sequence to provide the compounds of the invention or they may be used to synthesize fragments which are ,sub,sequently joined by the reactions described in the Reaction 10 Schemes.

REACTION SCHEME A
Reaction A. Couplin~ of re~sidues to form an amide bond N I OH + ~3 ~ R4b R4a EDC, HOBT H ~ CO2R
or HOOBT >~ R4b 2 J~ ~
R ~ Q

R4a~ R4b W 097136891 PCTrUS97/05707 REACTION SCHEME B

Reaction B. Preparation of reduced peptide ~ubunit.s by reductive alkylation ~ ~ R4b NaCNBH3 ~,~0~ N~zR

R4b / - -R4a REACTION SCHEME C

Reaction C. Deprotection of reduced peptide subunits >~O~N < TFA or O RA ~ Q \
~ R4b HCI
R4a H2N~ CO2R
- ~
RA ( Q~)_ ~C,,/ R4b R4a REACTION SCHEME D

Reaction D. Couplin~ of residues to form an amide bond EDC, HOBT

~oJ~ HN - R6b Et N DMF

R4b R4a O ~ R5a HCI orTFA
~OJJ~ N R5b Q \
~R4b R4a o H~ ~~ ~RR55ba ~Q ~
~R4b R4a W O97/36891 PCT~US97/05707 REACTION SCHEME E

Reaction E. Preparation of reduced dipeptides from peptides ~, ~ < BH3 THF
O RA ~Q ~
~R4b R4a >I' o RA ~ Q
)~R4b R4a where RA is R2, R3, R5a or RSb as previously defined; R4a and R4b are as previously defined; and R is an appropriate protecting group for the carboxylic acid.
Reaction Schemes F - M illustrate reactions wherein the 10 non-sulfhydryl-con~ining moiety at the N-terminus of the compounds of the instant invention is attached to an acyclic peptide unit which may be further elaborated to provide the instant compounds. It is well understood that such reactions are eclually useful when the - NHC(RA) - moiety of the reagents and compounds illustrated is replaced with the following 1 5 moiety:

((~ H2)t ~R7b R7a These reactions may be employed in a linear sequence to provide the compounds of the invention or they may be used to synthesize fragments W O 97/36891 PCT~US97tO5707 - 3~ -which are subse4uently joined by the reaction.s described in Reaction Schemes A - E.
The intermediates whose synthesis are illustrated in Reaction Schemes A and C can be reductively alkylated with a variety of aldehydes, such as I, as shown in Reaction Scheme F. The aldehydes can be prepared by standard procedures, such as that described by O. P. Goel, U. Krolls, M. Stier and S. Kesten in Organic Syntheses. 1988, 67, 69-75, from the appropriate amino acid (Reaction Scheme F). The reductive alkylation can be accomplished at pH 5-7 with a variety of reducing 10 agents, such as sodium triacetoxyborohydride or sodium cyanoboro-hydride in a solvent such as dichloroethane, methanol or dimethylforma-mide. The product II can be deprotected to give the final compounds III
with trifluoroacetic acid in methylene chloride. The final product III is isolated in the salt form, for example, as a trifluoroacetate, hydrochloride 15 or acetate salt, among others. The product diamine III can further be selectively protected to obtain IV, which can subsequently be reductively alkylated with a second aldehyde to obtain V. Removal of the protecting group, and conversion to cyclized products such as the dihydroimidazole VII can be accomplished by literature procedures.
Alternatively, the protected dipeptidyl analog intermediate can be reductively alkylated with other aldehydes such as l-trityl-4-carboxaldehyde or l-trityl-4-imidazolylacetaldehyde, to give products such as VIII (Reaction Scheme G). The trityl protecting group can be removed from VIII to give IX, or alternatively, VIII can first be treated 25 with an alkyl halide then subsequently deprotected to give the alkylated imidazole X. Alternatively, the dipeptidyl analog interrnediate can be acylated or sulfonylated by standard techniques.
The imidazole acetic acid XI can be converted to the acetate XIII by standard procedures, and XIII can be first reacted with an alkyl 30 halide, then treated with refluxing methanol to provide the regiospecific-ally alkylated imidazole acetic acid ester XIV. Hydrolysis and reaction with the protected dipeptidyl analog intermediate in the presence of condensing reagents such as 1-(3-dimethylaminopropyl)-3-ethylcarbo-diimide (EDC) leads to acylated products such as XV.

W O97136891 PCT~US97/05707 If the protected dipeptidyl analog intermediate is reductively alkylated with an aldehyde which also has a protected hydroxyl group, such as XVI in Reaction Scheme I, the protecting groups can be subse4uently removed to unmask the hydroxyl group (Reaction Schemes 1, J). The alcohol can be oxidized under standard conditions to e.g. an aldehyde, which can then be reacted with a variety of organometallic reagents such as Grignard reagents, to obtain secondary alcohols such as XX. In addition, the fully deprotected amino alcohol XXI can be reductively alkylated (under conditions described previously) with a 10 variety of aldehydes to obtain secondary amines, such as XXII (Reaction Scheme K), or tertiary amines.
The Boc protected amino alcohol XVIII can also be utilized to synthesize 2-aziridinylmethylpiperazines such as XXIII (Reaction Scheme L). Treating XVIII with l,l'-sulfonyldiimidazole and sodium 15 hydride in a solvent such as dimethylformamide led to the forrnation of aziridine XXIII . The aziridine reacted in the presence of a nucleophile, such as a thiol, in the presence of base to yield the ring-opened product XXIV .
In addition, the protected dipeptidyl analog intermediate can 20 be reacted with aldehydes derived from amino acids such as O-alkylated tyrosines, according to standard procedures, to obtain compounds such as XXX, as shown in Reaction Scheme M. When R' is an aryl group, XXX
can first be hydrogenated to llnm~;k the phenol, and the amine group deprotected with acid to produce XXXI. Alternatively, the amine 25 protecting group in XXX can be removed, and O-alkylated phenolic amines such as XXXII produced.
Similar procedures as are illustrated in Reaction Schemes F-M may be employed using other peptidyl analog intermediates such as those whose synthesis is illustrated in Reaction Schemes B - E.
Reaction Schemes N-R illustrate syntheses of suitably substituted aldehydes useful in the syntheses of the instant compounds 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.

REACTION SCHEME F

Boc NH~

2 ~ CO2R Boc NH CHO
R ~D Q ~ NaBH(OAc)3 ~ R4b Et3N, CICH2CH2CI
R a NHBoc ~_H Y CO R CF3co2H
Boc NH N~JI~ / 2 ,~f\ CH2CI2 R ~ Q ~

R4a~ R4b NH/~ N~JI~ ~02R Boc20 RA l~ Q ~ CH2CI2 R4a>~ R4b ~ H2 y CO2R ~CHO

BocN H N~\ <
- ~ NaBH(OAc)3 Q ) Et3N, CIC
IV ~ R4b R4a REACTION SCHEME F (continued) /=\
~, NH CF3CO2H, C H2CI2;
BocNH/~ NH~I~ CO2R NaHCO3 R ~ Q ~
~ R4a~ R4b NH

2 ~ ~ ~NC
RA ~ Q \ AgCN
Vl ~R4b R4a H Y
N~l~ CO2R

/~ RA ~ Q ~
N~,N~ '~,1~ R4b R4a Vll REACTION SCHEME G
y H2N J~ &o2R NaBH(OAc)3 Et3N, CICH2CH2CI
R
R4a~ R4b ~(CH2)nCHO

Tr H2)n+1 ,JI, <

~C ~ R
~4a Tr ~ ~ 1 ) Ar CH2X, CH3CN
VHI 2) CF3C02H, CH2C12 CF3CO2H, CH2CI2 (c2H5)3siH
(C2H5)3siH

(CH2)n~ ~\ ~'<
N ~ R4a~ R4b H IX

X R

W O97/36891 PCT~US97/05707 REACTION SCHEME H

N~ 2CO2H CH N_~CH2C02CH3 H HCI NH . HCI
Xl Xll N CH2C02CH31 ) ArCH2X CH3CN
(c6H5)3cBr ~ reflux (C2Hs)3N N 2) CH30H, retlux DMF Tr Xlll Ar--\N ~CH2c~2cH3 2.5N HClaq ~ 55~C
N

XIV

Ar~\N~C H2CO2H

N

PCTtUS97tO5707 REACTION SCHEME I

H2N ~ ~ 2 R4a~ R4b EDC HCI
HOBt DMF

Ar~ ~<~
XV R4a R4b W 097/36891 PCT~US97/OS707 REACTION SCHEME J
NaBH(OAc)3 CO2R Et3N, CICH2CH2CI
H2N J~ ( RA ~ Q \~ BnOl ~ R4b BocNH CHO
R4a XVI

NHBoc /~N~JI~ CO2R 20% Pd(OH)2 H2 BnO H J~l--< CH30H
( Q ~ CH3CO2H
XV I 1 4a ~ R4b R

NHBoc ~NJ~ CO2R CICOCOCI
HO H ,N ~ DMSO CH2C12 RA ( Q~) (C2H5)3N
\/ /--D 4b R 4a n PCTrUS97tO5707 REACTION SCHEME J (CONTINUED) H NHBoc y O~HN~J~ < (c2H6)~o RA ~ Q ~ 2. TFA, CH2C12 ~R4b XIX R4a/

R' NH2 HO>~-- Y CO R
RA /~Q ~
XX ~ R4b R4a WO97136891 PCT~US97/05707 REACTlON SCHEME K

NHBoc y CF3CO2H
HO/~H JI, CO2R CH2CI2 RA I~Q ~
XVIII ~__~ R4b R4a NH2 y R'CHO
~( 11 C02R
HO \~HN ~ < NaBH(OAc)3 ~) CICH2CH2CI
~/ ~ r~ 4b R4a n R'CH2~
NH y HO/~H~ CO2R

RA ~Q ~
XXII ~ R4b R4a W O 97136891 PCTrUS97/05707 - 4~ -REACTION SCHEME L

NHBoc H H
H~2R ~N~S,N~

XVIII R4b NaH, DMF 0~C
R4a RA Q< (C2Hs)3N
XXI I I ~ R4b C H30H
R4a NH2 y R'S/~H \J~ CO2R
RA ~Q ~
XXIV ~ R4b R4a PCTrUS97/05707 REACTION SCHEME M

HO~ 1) Boc20, K2C~3 HO,~

2) C H2N2, EtOAc ~
H2NCO2H BocNH CO2CH3 XXV XXVI

HO~
LiAlH4 ~1~ R'CH2X
TH F 1 Cs2CO3 0-20~C BocNH CH2OH DMF

XXVII

R'CH20,~ pyridine SO )~

DMSO
H (C2Hs)3N BocNH CHO
BocNH CH2O 20~C
XXVIII XXIX

CA 022496l7 l998-09-22 W O97/36891 PCTrUS97/05707 REACTION SCHEME M (continued) R'CH o~3 ~

BocNH CHO R4a R4b XXIX
NaBH(OAc)3 NHBoc R'CH20~ R

XXX R4b R4a 1) 20% Pd(OH)2 / HC~OAc CH30H, CH3C02H

R'CH 0~( ~R4b XXXI R4b R4a CA 022496l7 l998-09-22 W O 97/36891 PCTrUS97/05707 REACTION SCHEME N

CH3 1) HN02,Br2 ~CO2CH3 ~ 2) KMnO4 l ll H2N N~ 3) MeOH,H+ Br~'Nf ~\~\ MgCI R6 Zncl2lNicl2(ph3p)2 ~,C~2CH3 NaBH4 (excess) ~,CH20H

SO3-Py, Et3N ~CHO

DMSO N

PCT~US97/05707 REACTION SCHEME P

R6 [~
~CO2CH3 ~\MgCI ~CO2CH3 Zn, CuCN

NaBH4 ~ SO3Py, Et3N ~
(excess) [~,CH20H DMSO ~CHO

Br~ ~ ~CO2CH3 ~ \ 9 ¢ ~ ~3"CO2CH3 ZnC12, NiC12(Ph3P)2 N

(excess) ~ SO3 Py Et3N ¢~ CHO

W O 97/36891 PCTrUS97/05707 REACTION SCHEME O

Br~1. LDA, CO2 Br~

N2. MeOH, H+ N

~/\ MgCI 1~ CO2CH3 ZnCI2, Nicl2(ph3p)2 N
-NaBH4 (excess) ~OH S03 Py, Et3N

DMSO
N

CHO

WO 97/36891 PCT~US9710S707 REACTION SCHEME R

CO CH
1. LDA, CO2 ~ Br 2. (CH3)3SiCHN2 R6 ~ Br R6 ~

Zn, NiC12(Ph3P)2 Nl~co2cH3 excess NaBH4 ~1~ SO3Py, Et3N
N~CH20H DMSO

R6 3~

N~CHO

WO 97/36891 PCTrUS97105707 REACTION SCHEME S

Reaction S. Alkylation of an amide o O ~ R5a ~ Q + 5bR -X NDaMHF
~R4b R a O ~ R5a HCI 0, TFA
'1 11 I--N
~'0'~,~~~ R
Q ~
~R4b R4a o --N' R
- 5b ~ Q
~R4b R4a The instant compounds are useful as pharmaceutical agents for m~mm~ls, especially for humans. These compounds may be administered to patients for use in the treatment of cancer. Examples 10 of the type of cancer which may be treated with the compounds of this invention include, but are not limited to, colorectal carcinoma, exocrine pancreatic carcinoma, myeloid leukemias and neurological tumor.s. Such tumors may ari~se by mutations in the ) as genes themselves, muta~ions in the proteins that can regulate Ras formation (i.e., neurofibromin (NF- 1), 15 neu, scr, abl, lck, fyn) or by other mechanisms.

W O 97t36891 PCT~US97/05707 The compounds of the instant invention inhibit farnesyl-protein transferase and the farnesylation of the oncogene protein Ras.
The instant compounds may also inhibit tumor angiogenesis, thereby affecting the growth of tumors (J. Rak et al. Cance1 Research, 55:4575-5 45~0 (1995)). Such anti-angiogenesis properties of the instant compounds may also be useful in the treatment of certain forms of blindness related to retinal vascularization.
The compounds of this invention are also useful for inhibiting other proliferative diseases, both benign and malignant, lO wherein Ras proteins are aberrantly activated as a result of oncogenic mutation in other genes (i.e., the Ras gene itself is not activated by mutation to an oncogenic form) with said inhibition being accomplished by the administration of an effective amount of the compounds of the invention to a m~mm~l in need of such treatment. For example, a 15 component of NF-1 is a benign proliferative disorder.
The instant compounds may also be useful in the treatment of certain viral infections, in particular in the treatment of hepatitis delta and related viruses (J.S. Glenn et al. Science, 256:1331-1333 (1992).
The compounds of the instant invention are also useful in 20 the prevention of restenosis after percutaneous translllmin~l coronary angioplasty by inhibiting neointim~l formation (C. Indolfi et al. Natu~ e medicine, 1:541-545(1995).
The instant compounds may also be useful in the treatment and prevention of polycystic kidney disease (D.L. Schaffner et al.
25 American Journal of Pathology, 142:1051-1060 (1993) and ,3. Cowley, Jr. et al.FASEB Journal, 2:A3160 (1988)~.
The compounds of this invention may be ~r~ministered to m~mm~ls, preferably humans, either alone or, preferably, in combination with pharmaceutically acceptable carriers or diluents, optionally with 30 known adjuvants, such as alum, in a pharmaceutical composition, according to standard pharmaceutical practice. The compounds can be administered orally or parenterally, including the intravenous, intra-muscular, intraperitoneal, subcutaneous, rectal and topical routes of administration .

W O 97/36891 PCT~US97/05707 For oral use of a chemotherapeutic compound according to this invention, the selected compound may be admini.stered, for example, in the form of tablets or capsules, or a.s an aqueous solution or .suspen-sion. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch, and lubricating agents, such as magnesium .stearate, are commonly added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous 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, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared, and the pH of the solutions should be suitably adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled in order to render the preparation isotonic.
The present invention also encompasses a pharmaceutical composition useful in the treatment of cancer, comprising the a-lmini-stration of a therapeutically effective amount of the compounds of this invention, with or without pharmaceutically acceptable carriers or diluents. Suitable compositions of this invention include aqueous solutions comprising compounds of this invention and pharmacologically acceptable carriers, e.g., saline, at a pH level, e.g., 7.4. The ,solutions may be introduced into a patient's intramuscular blood-.stream by local bolus injection.
When a compound according to this invention is administered into a human .subject, the daily dosage will normally be determined by the prescribing physician with the dosage generally varying according to the age, weight, and response of the individual patient, as well as the severity of the patient's .symptoms.
In one exemplary application, a suitable amount of compound is ~lministered to a m~mm~l undergoing treatment for cancer.
Administration occurs in an amount between about 0.1 mg/kg of body weight to about 20 mgtkg of body weight per day, preferably of between 0.5 mg/kg of body weight to about 10 mg/kg of body weight per day.

WO 97136891 PCTrUS97105707 5~

The compounds of the instant invention are also useful as a component in an a~ssay to rapidly determine the presence and quantity of farnesyl-protein transferase (FPTa.se) in a composition.
Thus the composition to be tested may be divided and the two portions 5 contacted with mixtures which comprise a known substrate of FPTase (for example a tetrapeptide having a cysteine at the amine terminu~) and farnesyl pyrophosphate and, in one of the mixtures, 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 10 farnesylate the substrate, the chemical content of the assay mixtures may be determined by well known immunological, radiochemical or chromatographic techniques. Because the compounds of the instant invention are selective inhibitors of FPTase, absence or quantitative reduction of the amount of substrate in the assay mixture without the 15 compound of the instant invention relative to the presence of the unchanged substrate in the assay cont~ining the instant compound is indicative of the presence of FPTase in the composition to be tested.
It would be readily apparent to one of ordinary skill in the art that such an assay as described above would be useful in identifying 20 tissue samples which contain farnesyl-protein transferase and quantitating 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 series of samples composed of aliquots of a tissue extract containing an unknown amount of farnesyl-protein trans-25 ferase, an excess amount of a known substrate of FPTase (for examplea tetrapeptide having a cysteine at the amine terminus) 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 30 that has a Ki substantially smaller than the concentration of enzyme in the assay 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.

W O97/36891 PCTAUS97/0~707 EXAMPLES

Examples provided are intended to assist in a further under-standing of the invention. Particular materials employed, species and 5 conditions are intended to be further illustrative of the invention and not limitative of the reasonable scope thereof.
The standard workup referred to in the examples refers to solvent extraction and washing the organic solution with 10% citric acid, 10% sodium bicarbonate and brine as appropriate. Solutions were dried 10 over sodium sulfate and evaporated in vac~o on a rotary evaporator.

Preparation of N-l l -(3-[1 H-Imidazol-4-yl]propionyl)-pyrrolidin-2(S)-15 ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide Step A: Diethyl l-acetyl-5-hydroxy-3-ethylpyrrolidine-2,2-dicarboxylate Sodium (4.02 g, 0.175 mol) was dissolved in a stirred 20 solution of diethyl acetamidomalonate (235.4 g, 1.19 mol) in abs EtOH
(1.4 L) at ambient temperature under argon. The reaction mixture was cooled to 0~C, and trans-2-pentenal (100 g, 1.0~ mol) was added drop-wise maintaining the reaction temperature at <5~C. After the addition, the reaction was allowed to warm to room temperature, stirred for 4 h, 25 then quenched with acetic acid (28 mL). The solution was concentrated in vacuo, and the residue dissolved in EtOAc (1.5 L), washed with 10%
NaHCO3 solution (2 x 300 mL), brine, and dried (MgSO4). The solution was filtered and concentrated to 700 mL, then heated to reflux and treated with hexane (1 L). On cooling, the title compound precipitated and was 30 collected, mp 106 - 109~C. 1 H NMR (CD30D) ~ 5.65 (d, 1 H, J= 5 Hz), 4.1 - 4.25 (m, 4H), 2.7-2.~s (m, lH), 2.21 (s, 3H), 2.10 (dd, lH, J = 6, 13, Hz),1.~6- 1.97 (m, 2H), 1.27 (t, 3H, J= 7 Hz), 1.23 (t, 3H, J= 7 Hz), 1.1-1.25 (m, lH)~ 0.97 (t, 3H, J= 7 Hz).

CA 022496l7 l998-09-22 W O 97/36891 PCT~US97/05707 Step B: Diethyl l -acetyl-3-ethylpyrrolidine-2,2-dicarboxylate To a solution of diethyl l-acetyl-S-hydroxy-3-ethyl-pyrrolidine-2,2-dicarboxylate (287 g, 0.95 mol) and triethylsilane (228 mL, 1.43 mol) in CH2C12 (3 L) under argon was added trifluoroacetic acid (735 mL, 9.53 mol) dropwise with stirring while maintaining the internal temperature at 25 ~C by means of an ice bath. After stirring for 3 h at 23~C, the solution was concentrated in vacuo,, the residue diluted with CH2C12 (1.5 L), then treated with H2O (1 L) and solid Na2CO3 with vigorou.s stirring until the solution was basic. The organic layer was ~separated, dried (Na2SO4), filtered, then concentrated to give the title compound as a yellow oil which was used without further purification.

Step C: 3-Ethylproline hydrochloride (Cis:Trans Mixture) Diethyl 1-Acetyl-3-ethylpyrrolidine-2,2-dicarboxylate (373 g, 0.95 mol) was suspended in 6N HCI (2 L) and HOAc (500 mL) and heated at reflux for 20 h ~e reaction mixture was cooled, washed with EtOAc (lL), then concentrated in vacuo to give an oil which crystallized upon trituration with ether to give the title compound. lH N~IR (D20) ~
4.23 (d, lH, J- 8 Hz), 3.84 (d, lH, J= 8 Hz), 3.15- 3.4 (m, 4H), 2.33- 2.44 (m, lH), 2.19-2.4 (m, lH), 2.02- 2.15 (m, 2H), 1.53- 1.72 (m, 3H), 1.23-1.43 (m, 2H), 1.0- 1.15 (m, IH), 0.75 - 0.83 (m, 6H).

Step D: N-[(tert-Butyloxy)carbonyl]-c~is:trans-3-ethylproline methyl ester 3-Ethylproline hydrochloride (Cis:Trans Mixture) (20 g, 0.11 mol) was dissolved in CH30H (200 mL), and the solution was saturated with HCI gas, then stirred at 23~C for 24 h. Argon was bubbled through the solution to remove excess HCl. The .solution was treated with NaHCO3 (>84 g) to a pH of 8, then di-tert-butyl dicarbonate (25.1 g, 0.115 mol) dissolved in CH30H (20 mL) was added slowly. After stirring for 18 h at 23~C, the mixture wa~s filtered, the filtrate concen-trated, and the residue triturated with EtOAc, filtered again, and concentrated to give the title compound as an oil.

W O97/36891 PCTrUS97/05707 Step E: N-[(te) t-Butyloxy)carbonyl]-tran.~-3-ethylproline and N-I (ter~-Butyloxy)carbonyll-cis-3-ethylproline methyl e~ster N-l(tert-Butyloxy)carbonyl]-c~is,t)~ans-3-ethylproline methyl ester (29.1 g, 0.113 mol) was dissolved in CH30H (114 mL) with cooling 5 to 0~C, then treated with 1 N NaOH (114 mL). After stirring for 20 h at 23~C, the solution was concentrated to remove the CH30H and then extracted with EtOAc (3 x). The organic layers were combined, dried (MgSO4), filtered, and concentrated to give 12.8 g of N-[(tert-Butyloxy)carbonyl]-cis-3-ethylproline methyl ester as an oil. The 10 aqueous layer was acidified with solid citric acid and extracted with EtOAc (2 x), the organic layers combined, dried (MgSO4), filtered, and concentrated to give N-[(tert-Butyloxy)carbonyl]-trans-3-ethylproline a~s an oil. lH NMR (CD30D) â 3.86 and 3.78 (2 d, lH, J = 6 Hz), 3.33 -3.58 (m, 2H), 2.01 - 2.22 (m, 2H), 1.5 - 1.74 (m, 2H), 1.33 - 1.5 (m, lH), 1.45 and 1.42 (2 s, 9H), 0.98 (t, 3H, J= 8 Hz).

Step F: 3(S)-Ethyl-2(S)-proline hydrochloride N-[(tert-Butyloxy)carbonyl]-t~ans-3-ethylproline (15.5 g, 0.064 mol), S-a-methylbenzylamine (9.03 mL, 0.070 mol), HOBT (10.73 20 g, 0.70 mol), and N-methylmorpholine (8 mL, 0.076 mol) were dissolved in CH2C12 (150 mL) with stirring in an ice-H2O bath, treated with EDC
(13.4 g, 0.070 mol) stirred at 23~C for 48 h. The reaction mixture was partitioned between EtOAc and 10% citric acid solution, the organic layer washed with saturated NaHCO3 solution, brine and dried (MgSO4), 25 filtered, and concentrated to give an oil. This oil was dissolved in a minimllm amount of ether (10 mL) to crystallize the desired S,S,S
diastereomer (4.2 g), mp 118- 121 ~C. A solution of this product in 8N
HCl (87 mL) and glacial acetic acid (22 mL) was heated at reflux overnight. The solution was concentrated on a rotary evaporator, and the 30 residue taken up in H20 and extracted with ether. The aqueous layer was concentrated to drynes.s to give a 1: 1 mixture of 3(S)-ethyl-2(S)-proline hydrochloride and oc-methylbenzylamine.
3(S)-Ethyl-2(S)-proline containing oc-methylbenzylamine (2.0 g, 0.0128 mol) was dissolved in dioxane (10 mL) and H2O (10 mL) CA 022496l7 l998-09-22 W O97/36891 PCT~US97/05707 with stirring and cooling to 0~C. N,N-diisopropylethylamine (2.2 mL, 0.012~S mol) and di-te~t-butyl-dicarbonate (2.79 g, 0.012~ mol) were added and stirring was continued at 23~C for 4~ h. The reaction mixture was partitioned between EtOAc (60 m~) and H2O (30 mL), the organic layer washed with 0.5N NaOH (2 x 40 mL), the aqueous layer~
combined and washed with EtOAc ( 30 mL) and this layer back-extracted with 0.5 N NaOH (30 mL). The aqueous layers were combined and care-fully acidified at 0~C with 1 N HCI to pH 3. This mixture was extracted with EtOAc (3 x 40 mL), the organ~cs combined, dried (MgSO4), filtered and concentrated to give N-[(tert-Butyloxy)carbonyl-3(S)-ethyl-2(S)-proline as a colorless oil. N-[(tert-Butyloxy)carbonyl-3(S)-ethyl-2(S)-proline was dissolved in EtOAc (50 mL) and the solution was saturated with HCI gas with cooling in an ice-H2O bath. The solution was stoppered and stirred at 0~C. for 3 hr. Argon was bubbled through the solution to remove excess HCI, and the solution wa~s concentrated to dryne,ss to give 3(S)-ethyl-2(S)-proline hydrochloride.

Step G: N-~(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline 3(S)-Ethyl-2(S)-proline hydrochloride (from Example 1, Step F) (2.33 g, 0.013 mol) was dissolved in CH30H (20 mL), treated with 3A molecular sieve~s (2 g) and KOAc (1.27 g, 0.013 mol) to adjust the pH of the reaction mixture to 4.5-5, then N-[(te1 t-Butyloxy) carbonyl-prolinal (Pettit et al., J. Org. Chem. (1994) 59, L21] 6287-95) (3.36 g, 0.017 mol) was added, and the mixture was stirred for 16 hours at room temperature. The reaction mixture was filtered, quenched with aqueous saturated NaHCO3 (5 mL) and concentrated to dryness. The residue was extracted with CHC13. The extract was dried (MgSO4), filtered, and concentrated to give the title compound and inorganic .salt~.

Step H: N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl] -3(S)-ethyl-proline (0.50 g, 1.53 mmol), EDC (0.293 g, 1.53 mmol), HOBT

CA 022496l7 l998-09-22 W O97/36891 PCTrUS97/05707 (0.243 g, 1.53 mmol) and 3-chlorobenzylamine(0.1~7 mL, 1.53 mmol) were di.ssolved in DMF (5 mL), the pH adjusted to 7 with N-methyl-morpholine (0.51 mL, 4.6 mmol), and the reaction mixture stirred for 1 ~ hours at ambient temperature. After removing the solvent iM l ac~uo 5 the residue was partitioned between EtOAc and 5% a4ueous NaHCO3.
The organic layer was washed with brine, dried (MgSO4), filtered, and concentrated to give the title compound after chromatography (SiO2, EtOAc: hexane, 2:3).
~0 Step I: N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl] -3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide (0.175 g, 0.39 mrnol) was dissolved in dry DMF (4 mL) with stirring at 0~C under Ar, treated with 15 NaH (60% dispersion in mineral oil, 0.023 g, 0.5~ mmol), and after 15 minutes treated with iodomethane (0.029 mL, 0.47 mmol). The reaction mixture was stirred at 25 ~C. for 2 h, then evaporated to dryness and partitioned between EtOAc and aqueous saturated NaHCO3 solution.
The organic layer was separated, washed with brine, dried (MgSO4), 20 filtered, and concentrated to dryness to give the title compound.

Step J: (Pyrrolidin-2(S)-ylmethyl)-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethvl) amide N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl] -3(S)-25 ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide (0.150 g, 0.32 mmol) was dissolved in EtOAc (15 mL), cooled to -20~C. and saturated with HCI gas. The .solution was stirred at 0~C. for 1 h, then at 25~C. for 1 hour, then concentrated to dryness to give pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide which 30 was used without further purification.

Step K: N-[ 1-(3-[1 H-Irnidazol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl] -3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide (Pyrrolidin-2(S)-ylmethyl)-3(S)-ethyl-proline-N-methyl-N-5 (3-chlorophenylmethyl) amide (0.070 g, 0.162 mmol), lH-imidazol-4-yl]propionic acid hydrochloride (0.057 g, 0.324 mmol), EDC (0.062 g, 0.324 mmol), HOBT (0.050 g, 0.324 mmol), and N-methylmorpholine (0.288 mL, 1.30 mmol) were dissolved in DMF (5 mL) at 25~C. and stirred for 72 h. The reaction mixture was partitioned between EtOAc 10 and 5% Na2CO3 solution, the organic layer separated, washed with brine, dried (MgSO4), filtered, and concentrated to dryness to give the title compound after preparative RP HPLC (Vydac column, 0.1 %
TFA/CH3CN: 0.1% TFA/H20, 95:5 to 5:95 gradient) and Iyophilization.
to give the title compound as the TFA salt.~5 Anal. calcd for C26H36N5O2CI ~ 1.9 CF3C02H- 2.0 H2O:
C, 48.45; H, 5.72; N, 9.48;
found: C, 48.53; H, 5.76; N, 9.08.
FAB MS 486 (M+l) Preparation of N-[ 1-(3-[ l H-Imidazol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide Step A: N-l(tert-Butyloxy)carbonyll-3(S)-ethyl-2(S)-prolinol 3(S)-Ethyl-2(S)-proline hydrochloride (Example 1, Step G) cont~ining oc-methylbenzylamine (2.0 g, 0.0128 mol) was dissolved in dioxane ( 10 mL) and H2O (10 mL) with stirring and cooling to 0~C.
30 N,N-diisopropylethylamine (2.2 rnL, 0.0128 mol) and di-tert-butyl-dicarbonate (2.79 g, 0.0128 mol) were added and stirring was continued at 23~C for 48 h. The reaction mixture was partitioned between EtOAc (60 mL) and H2O (30 mL), the organic layer washed with 0.5N NaOH (2 x 40 mL), the aqueous layers combined and washed with EtOAc ( 30 mL) 35 and this layer back-extracted with 0.5 N NaOH (30 mL). The aqueous layers were combined and carefully acidified at 0~C with lN HCI to pH

WO 97/36891 PCT~US97/05707 2. This mixture was extracted with EtOAc (3 x 40 mL), the organics combined, dried (MgSO4), filtered and concentrated to give N-[(te~ t-Butyloxy)carbonyl--3(S)-ethyl-2(S)-proline as a colorless oil which was used without purification.

N-[(tert-Butyloxy)carbonyl]-3(S)-ethyl-2(S)-proline (1.6 g, 6.5~ m~mol) was dissolved in dry THF (10 mL) and treated with borane (lM in THF, 12.5 mL, 12.5 mmol) with stirring at 0 ~C for 2 h, then 23~C for 1 h. The solution was cooled to 0~C, treated with H2O (20 mL), and extracted 10 with EtOAc (2 x 30 mL). The organics were washed with brine, saturated NaHCO3, H2O, dried (MgSO4), filtered and concentrated to give a viscous oil. The oil was dissolved in CH2C12, filtered through dry SiO2, and the filtrate concentrated to give the title compound as an oil.
lH NMR (CDC13) o 4.97 (d, lH, J= 7 Hz), 3.71 (t, lH, J = 8 Hz), 3.51-15 3.62 (m, 3H), 3.1~ - 3.26 (m, lH), 1.9 - 2.0 (m, lH), 1.53-1.7 (m, 2H), 1.47 (s, 9H), 1.26 - 1.43 (m, 2H), 0.95 (t, 3H, J = 7 Hz).

Step B: N-~(tert-Butyloxy)carbonyll-3(S)-ethvl-2(S)-prolinal N-[(te1 t-Butyloxy)carbonyl-3(S)-ethyl-2(S)-prolinol 20 (0.63~ g, 2.78 mmol) and Et3N (1.4 mL, 9.74 mmol) were dissolved in dry CH2C12 (10 mL) with stirring and cooling to -10~C and treated dropwise with a solution of S03.pyr (1.33 g, 8.35 mmol) in dry DMSO
(5 mL) keeping the reaction mixture temperature at < 0~C. The mixture was stirred at 0~C. for 20 minutes then at 5~C for 20 minlltes, and at 25 1 5~C for 1 hour, then poured into ice-cold 0.5 N HCI and the layers separated. The aqueous layer was extracted with CH2C12 (3 x 20 mL), organics combined, washed with H2O, aqueous saturated NaHCO3 solution, brine, and dried (Na2SO4). Filtration and concentration to dryness gave the title compound which was used without purification.

Step C: N-[(t-Butyloxycarbonyl)-3(S)-ethylpyrrolidin-2~S)-ylmethyll -3(S)-ethyl-proline Following the procedure outlined in Example 1, Step G, but substituting N-[(tert-butyloxy)carbonyl]-3(S)-ethyl-2(S)-prolinal for N-[(tert-Butyloxy)carbonyl]-2(S)-prolinal the title compound was prepared.

Step D: N-[ l -(3-[1 H-Imidazol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-ylmethyll-3(S)-ethyl -proline-N-methyl-N-(3-chloro-phenylmethyl) amide Using the procedures de~scribed in Example 1, the title compound is prepared.

I ~S Preparation of 1 -~ 1 -(4-Cyanobenzyl)- 1 H-imidazol-5-ylacetyl] pyrrolidin-2(S)-ylmethyl] -3 (S)-ethyl-proline-N-methyl-N-(3 -chloro-phenylmethyl) amide Step A: lH-lmidazole-4- acetic acid methyl ester hydrochloride A solution of lH-imidazole-4-acetic acid hydrochloride (4.00g, 24.6 mmol) in methanol (100 ml) was saturated with gaseous hydrogen chloride. The resulting solution wa~s allowed to stand at room temperature (RT) for 1 ~ hours. The solvent was evaporated in vacuo to afford the title compound as a white solid.
lH NMR(CDC13, 400 MHz) ~ 8.85(1H, s),7.45(1H, s), 3.~¢9(2H, s) and 3.75(3H, s) ppm.

Step B: 1 -(Tribenzyl)- 1 H-imidazol-4-ylacetic acid methyl ester To a solution of 1 H-Imidazole-4- acetic acid methyl ester hydrochloride (24.85g, 0.141mol) in dimethyl formamide (DMF) (l l5ml) was added triethylamine (57.2 ml, 0.412mol) and tribenzyl - bromide(S5.3g, 0.171 mol) and the suspension was stirred for 24 hours.
After this time, the reaction mixture was diluted with ethyl acetate (EtOAc) (I 1) and water (350 ml). The organic phase was washed with W O 97/36891 PCTrUS97/05707 saturated. a4ueous. NaHCO3 (350 ml), dried (Na2SO4) and evaporated in vacuo. The residue was purified by flash chromatography (sio2~ 0-100% ethyl acetate in hexanes; gradient elution) to provide the title compound a,s a white solid.
lH ~MR (CDC13, 400 MHz) o 7.35(1H, s), 7.31 (9H, m),7.22(6H, m), 6.76(1H, s), 3.6~(3H, .s) and 3.60(2H, s) ppm.

Step C: 11-(4-Cyanobenzyl)-lH-imidazol-5-yllacetic acid methyl ester To a solution of l-(Tribenzyl)-lH-imidazol-4-ylacetic acid methyl ester (~.00g, 20.9mmol) in acetonitrile (70 ml) was added bromo-p-toluonitrile (4.10g, 20.92 mmol) and heated at 55~C for 3 hr. After this time, the reaction was cooled to room temperature and the resulting imidazolium salt (white precipitate) was collected by filtration. The filtrate was heated at 55~C for l~s hours. The reaction mixture was cooled to room temperature and evaporated in vacuo. To the residue was added EtOAc (70 ml) and the resulting white precipitate collected by filtration. The precipitated imidazolium salts were combined, suspended in methanol (100 ml) and heated to reflux for 30 minutes. After this time, the solvent was removed in vacuo, the resulting residue was suspended in EtOAc (75ml) and the .solid isolated by filtration and washed (EtOAc).
The solid was treated with saturated aqueous NaHCO3 (300ml) and CH2C12 (300ml) and stirred at room temperature for 2 hr. The organic layer was separated, dried (MgSO4) and evaporated in vacuo to afford the title compound as a white solid:
1HNMR(CDC13, 400 MHz) ~ 7.65(1H, d, J=~Hz), 7.53(1H, s), 7.15(1H, d, J=f~Hz), 7.04(1H, s), 5.24(2H, s), 3.62(3H, s) and 3.45(2H, s) ppm.

Step D: ~ 1 -(4-Cyanobenzyl)- 1 H-imidazol-5-yllacetic acid A solution of [ I -(4-cyanobenzyl)- 1 H-imidazol-5-yl]acetic acid methyl ester (4.44g, 17.4mmol ) in THF (lOOml) and 1 M lithium hydroxide (17.4 ml, 17.4 mmol) was stirred at RT for 1 ~ hr. I M HCI
(17.4 ml) was added and the THF was removed by evaporation in vacuo.

W O 97/36891 PCT~US97/05707 The a4ueous ~solution wa~s Iyophilized to afford the title compound containing lithium chloride as a white ~solid.
1 H NMR(CD30D, 400 MHz) ~ 8.22(1 H, s), 7.74(1 H, d, J-8.4Hz), 7.36(1H, d, J=8.4Hz), 7.15(1H, s), 5.43(2H, s) and 3.49(2H, s) ppm.
s Step E: I -[1 -(4-Cyanobenzyl)- 1 H-imidazol-5-ylacetyl]
pyrrolidin-2(S )-ylmethyl] -3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide Pyrrolidin-2(S)-ylmethyll -3(S)-ethyl-proline-N-methyl-N-10 (3-chlorophenylmethyl) amide (0.070 g, 0.1621nmol) (Example 1, Step J), [1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetic acid ~ LiCI (0.093 g, 0.324 mmol), EDC (0.062 g, 0.324 mmol), HOBT (0.050 g, 0.324 mmol), and N-methylmorpholine (0.288 mL, 1.30 mmol) were dissolved in DMF
(5 mL) at 25~C. and stirred for 72 h. The reaction mixture was parti-15 tioned between EtOAc and 5~O Na2C03 solution, the organic layerseparated, washed with brine, dried (MgSO4), filtered, and concentrated to dryness to give the title compound after preparative RP HPLC (Vydac column, 0.1 % TFA/CH3CN: 0.1 % TFA/H2O, 95:5 to 5:95 gradient) and Iyophilization. to give the title compound as the TFA salt. lH NMR
20 (CD30D, 400 MHz) ~ 8.96 and 8.93 (lH, 2 s, 1:2 ratio), 7.7~ - 7.85 (2H, m), 7.5 - 7.62 (3H, m), 7.2 - 7.4 (4H, m), 5.60 and 5.67 (2H, 2 s, 1 :2 ratio), 4.55 - 4.8 (2H, m), 4.09 - 4.2 (lH, m), 3.82 -4.0 (3H, m), 3.5 - 3.65 (2H, m), 3.35 - 3.49 (2H, m), 3.1~ - 3.3 (lH, m), 3.10 (3H, s), 2.1 - 2.4 (4H, m), 1.92 - 2.09 (3H, m), 1.71 - 1.92 (2H, m), 1.5 - 1.7 (lH, m), 0.96 25 - 1.08 (3H, m) ppm.
FAB MS 587 (M+l ) 30 Preparation of 1 -[1 -(4-Cy~nobenzyl)- 1 H-imidazol-S-ylacetyl] pyrrolidin-2(S)-ylmethyl] -3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide WO 97/36891 PCT~US97105707 Step A: (Pyrrolidin-2(S)-ylmethyl)-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide N-[(t-Butyloxycarbonyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide (Example 1, Step H) 5 (0.175 g, 0.39 mmol) was dissolved in EtOAc (15 mL), cooled to -20~C.
and ~aturated with HCI gas. The solution was ~tirred at 0~C. for 1 h, then at 25~C. for 1 h, then concentrated to dryness to give pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide which wa,<;
used without further purification.
Step B: 1 -[ I -(4-Cyanobenzyl)- 1 H-imidazol-5-ylacetyl]
pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethvl) amide Using the procedures described in Example 3, but substituting pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide for pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide in Step E, the title compound was prepared. lH NMR (CD30D, 400 MHz) ~ 8.94 (lH, ~), 7.~s1 (2H, d, J= S Hz), 7.5 - 7.6 (3H, m), 7.12 - 7.3X (4H, m), 5.55 (2H, s), 4.38 - 4.54 (2H, m), 4.02 - 4.1~ (lH, m), 3.75 -4.01 (4H, m), 3.42 - 3.65 (2H, m), 3.42 - 3.65 (2H, m), 3.1 - 3.27 (lH, m), 2.2 - 2.4 (2H, m), 1.6 -2.2 (7H, m), 1.42 - 1.5~ (lH, m), 1.22 - 1.35 (lH, m), 0.95 - 1.0~ (3H, m) ppm.
FAB MS 573 (M+l) Preparation of 1 -(1 -(4-Nitrobenzyl)- 1 H-imidazol-4-ylacetyl]
pyrrolidin -2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide and 1-[1-(4-Nitrobenzyl)-lH-imidazol-5-ylacetyl] pyrrolidin -2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethvl) amide CA 022496l7 l998-09-22 W O 97/36891 PCTrUS97/OS707 Step A: 1-(4-Nitrobenzyl)-lH-imidazol-4-ylacetic acid methyl ester and 1 -(4-Nitrobenzyl)- I H-imidazol-5-ylacetic acid methyl ester (3:1 mixture) To a solution of sodium hydride (60% in mineral oil, 99 mg, 2.5 mmol) in dimethylformamide (2 ml) cooled to 0~C was added, via cannula, a solution of lH-imidazole-4-acetic acid methyl ester hydrochloride (200mg, 1.13 mmol) in dimethylformamide (3 ml). This suspension was allowed to stir at 0~C for 15 min. To this suspension wa,s added 4-nitrobenzyl bromide (244 mg, 1.13 mmol) and stirred at room temperature for 2 h. After this time, the mixture was quenched with saturated. aqueous. sodium bicarbonate (15 ml) and water (20 ml) and extracted with methylene chloride (2 x 50 ml). The combined organic extracts were washed with brine (20 ml), dried (MgSO4), filtered and the solvent was evaporated in vacuo. The residue was purified by flash chromatography using acetonitrile as eluent to give the title compounds as a yellow oil.
lH NMR (CDC13, 400 MHz) ~ 8.20 (2H, d, J=8.5 Hz), 7.49 (lH, s), 7.27 (2H, d, J=8.5 Hz), 7.03 (0.25H, s), 6.87 (0.75H, s), 5.28 (O.SH, s), 5.18 (1.5H, s), 3.70 (2.25H, s), 3.65 (1.5H, s), 3.61 (0.75H? s) and 3.44 (0.5H, 20 s) ppm.

Step B: 1-(4-Nitrobenzyl)-lH-imidazol-4-ylacetic acid hydrochloride and I -(4-Nitrobenzyl)- lH-imidazol-S-ylacetic acid (3:1mixture) To a solution of a mixture of I-(4-Nitrobenzyl)-lH-imidazol-4-ylacetic acid methyl ester and 1-(4-Nitrobenzyl)-lH-imidazol-5-ylacetic acid methyl ester (3:1mixture, 216 m~, 0.785 rnrnol) in methanol (3 ml) and tetrahydrofuran (3 ml) under argon was added 1.0 M sodium hydroxide (1.18 ml, 1.18 mmol) and stirred for 1~ h. After this time, 1.0 N hydrochloric acid (2.36 ml, 2.36 mmol) was added and the mixture evaporated in vacuo to give the title compounds.
lH NMR (CDC13, 400 MHz) ~ 9.04 (0.75H, s), ~.83 (0.25H, s), 8.28 (2H, d, J=8.8 Hz), 7.61 (2H, d, J=8.~ Hz), 7.54 (0.75H, s),7.43 (0.25H?
s), 5.61 (0.5H, s), 5.58 (1.5H, s), 3.84 (0.5H, s) and 3.8~ (1.5H? s) ppm.

W O97/36891 PCTrUS97105707 Step C~ (4-Nitrobenzyl)- 1 H-imidazol-4-ylacetyl]
pyrrolidin -2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide and 1-[1-(4-Nitrobenzyl)-lH-imidazol-5-ylacetyl] pyrrolidin -2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide Using the procedure.~ outlined in Examples 3 and 4, the title compound~ are prepared.

The following compounds are prepared in a ~imilar manner:
I -(1 -(4-Nitrobenzyl)- 1 H-imidazol-4-ylacetyl] pyrrolidin -2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide 1-[1 -(4-Nitrobenzyl)- 1 H-imidazol-5-ylacetyll pyrrolidin -2(S)-ylmethyll-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide -1 -(1 -(4-Methoxybenzyl)- 1 H-imidazol-5-ylacetyl)pyrrolidin-2(S)-ylmethyll-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide 20 1-(1 -(4-Methoxybenzyl)- 1 H-imidazol-5-ylacetyl)pyrrolidin-2(S)-vlmethyll-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide 1-(1 -(2-Naphthylmethyl)- 1 H-imidazol -5-ylacetyl]pyrrolidin-2(S)-ylmethyll-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide 1-(1 -(2-Naphthylmethyl)- 1 H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyll-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide Preparation of 1-(1-(1 -Farnesyl)- 1 H-imidazol-S-ylacetyl)-pyrrolidin-2(S)-ylmethyll -3(S)-ethyl -proline-N-methyl-N-(3-chlorophenylmethyl) amide Step A: l-(1-Farnesyl)-lH-imidazol-5-ylacetic acid methyl e,ster To a solution of l-(tribenzyl)-lH-imidazol-4-ylacetic acid methyl e.ster (200 mg, 0.523 mmol) in acetonitrile (5 ml) was added trans, trans-farnesyl bromide (156 ,ul, 0.575 mmol) and heated at 55~C for 16 h.
5 After this time, the reaction was heated at 80~C for 3 h and then the reaction mixture was evaporated in vacuo. The residue was dissolved in methanol (5 ml ) and heated to reflux for 30 minutes and then evaporated in vacuo. The residue was purified by flash chromatography (2-4%
methanol/methylene chloride gradient elution) to provide the title 10 compound.
lH NMR (CDC13, 400 MHz) ~ 7.50 (lH, s), 6.92 (lH, s), 5.24 (lH, t, J-5.9 Hz), 5.09 (2H, m), 4.49 (2H, d, J=6.9 Hz), 3.69 (3H, s), 3.60 (2H, s), 1.91-2.15 (8H, m), 1.72 (3H, s), 1.65 (3H, s), 1.59 (3H, s) and 1.57 (3H, s) ppm.
Step B: 1 -(1 -(1 -Farnesyl)- 1 H-imidazol-S-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide Following the procedure described in Example 5, but using 20 1 -farnesyl- 1 H-imidazol-5-ylacetic acid methyl ester described in Step A
in place of 1-(4-nitrobenzyl)-lH-imidazol-S-ylacetic acid methyl ester provides the title compound.

1 -(1 -(1 -Geranyl)- 1 H-imidazol-5-ylacetyl)-pyrrolidin-2(S)-ylmethyl] -25 3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide is prepared in a similar manner.

30 In vitro inhibition o~ ras farnesyl transfera.se Assays offarnesyl-protein tran~èrase. 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.
2~5: 14701 - 14704 (1990), Pompliano, et al., Biochemist~y 31 :3800 W O 97/36891 PCT~US97105707 ( l 992) and Gibbs et al., PNAS U.S.A. 8~:6630-6634 ( l 9g9), respectively.
Bovine FPTase wa.s assayed in a volume of l00 !ll containing l00 mM
N-(2-hydroxy ethyl) piperazine-N'-(2-ethane .~iulfonic acid) (HEPES), pH
7.4, 5 mM MgCl2, 5 mM dithiothreitol (DTT), l00 mM [3H]-farnesyl 5 diphosphate ([3H]-FPP; 740 CBq/mmol, New ~ngland Nuclear), 650 nM
Ras-CVLS and 10 ~g/ml FPTase at 31 ~C for 60 min. Reaction,s were initiated with FPTase and stopped with 1 ml of l.0 M HCL in ethanol.
Precipitates were collected onto filter-mats using a TomTec Mach II cell harvestor, washed with 100% ethanol, dried and counted in an LKB
l0 ~-plate counter. The assay was linear with respect to both substrates, FPTase levels and time; less than 10% of the ~3H]-FPP was utilized during the reaction period. Purified compounds were dissolved in 100%
dimethyl sulfoxide (DMSO) and were diluted 20-fold into the assay.
Percentage inhibition is measured by the amount of incorporation of l 5 radioactivity in the presence of the test compound when compared to the amount of incorporation in the absence of the test compound.
Human FPTase was prepared as described by Omer et al., Biochemistry 32:5167-5l76 (1993). Human FPTase activity was assayed as described above with the exception that 0.1% (w/v) polyethylene 20 glycol 20,000, 10 ~lM ZnCl2 and l00 nM Ras-CVIM were added to the reaction mixture. Reactions were performed for 30 min., stopped with 100 !11 of 30% (v/v) trichloroacetic acid (TCA) in ethanol and processed as described above for the bovine enzyme.
The compounds of the instant invention were tested for 25 inhibitory activity against human FPTase by the assay described above and were found to have IC50 of < 10 ~M.

EXAMPLE

30 In viv(~ ras farnesylation assay The cell line used in this assay is a v-ras line derived from either Ratl or NIH3T3 cell~;, which expressed viral Ha-ras p21. The assay is performed essentially as described in DeClue, J.E. et al..

W O97/36891 PCTrUS97/05707 CancerResea~ch 51:712-717, (1991). Cell.s in 10 cm dishes at 50-75%
confluency are treated with the test compound (final concentration of solvent, methanol or dimethyl sulfoxide, is 0.1 %). After 4 hours at 37~C, the cells are labeled in 3 ml methionine-free DMEM supple-meted with 10% regular DMEM, 2% fetal bovine serum and 400 mCiL35S]
methionine (1000 Ci/mmol). After an additional 20 hours, the cells are Iysed in 1 ml Iysis 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) and the Iysates cleared by centrifugation at 100,000 x g for 45 min. Aliquots of Iysates containing equal numbers of acid-precipitable counts are bought to I ml with IP buffer (Iysis buffer lacking DTT) and immunoprecipitated with the ras-specific monoclonal antibody Y13-259 (Furth, M.E. etal., J. Vi~ol. 43:294-304, (19~2)).
Following a 2 hour antibody incubation at 4~C, 200 ml of a 25%
suspension of protein A-Sepharose coated with rabbit anti rat IgG is added for 45 min. The immunoprecipitates are washed four times with IP buffer (20 nM HEPES, pH 7.5/1 mM EDTA/1 % Triton X- 100Ø5%
deoxycholate/0.1 %/SDS/0.1 M NaCI) boiled in SDS-PAGE sample buffer and loaded on 13% acrylamide gels When the dye front reached the bottom, the gel is fixed, soaked in Enlightening, dried and auto-radiographed. The intensities of the bands corresponding to farnesylated and nonfarnesylated ras protein~s are compared to determine the percent inhibition of farnesyl transfer to protein.

ln vil~o ~rowth inhibition assay To determine the biological conse4uence~s of FPTase inhibition, the effect of the compounds of the instant invention on the 30 anchorage-independent growth of Ratl cells transformed with either a v-ras, v-~ af, or v-mos oncogene is tested. Cells transforrned by v-Raf and v-Mos maybe included in the analysis to evaluate the specificity of instant compounds for Ras-induced cell transformation.
Rat 1 cells transformed with either v-ras, v-raf, or v-mos CA 022496l7 l998-09-22 W O97/36891 rCTAUS97/05707 are seeded at a den.sity of 1 x 104 cells per plate (35 mm in diameter) in a 0.3% top agarose layer in medium A (Dulbecco'~s modified Eagle's medium supplemented with 10% fetal bovine serum) over a bottom agarose layer (0.6%). Both layers contain 0.1% methanol or an 5 appropriate concentration of the instant compound (dis~solved in methanol at 1000 times the final concentration used in the assay). The cells are fed twice weekly with 0.5 ml of medium A containing 0.1% methanol or the concentration of the instant compound. Photomicrographs are taken 16 days after the cultures are seeded and comparisons are made.

Claims (24)

WHAT IS CLAIMED IS:

1. A compound which inhibits Ras farnesyl-transferase having the Formula I:

wherein:

R1a and R1b are independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, alkenyl, alkynyl, R10O, R11S(O)m, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, N3, -N(R10)2, or R11OC(O)NR10-, -C(O)N(R10)2 c) C1-C6 alkyl unsubstituted or substituted by aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, N3, -N(R10)2, or R11OC(O)-NR10-;

R2 and R3 are independently selected from:
a) a side chain of a naturally occurring amino acid, b) an oxidized form of a side chain of a naturally occurring amino acid which is:
i) methionine sulfoxide, or ii) methionine sulfone, and c) substituted or unsubstituted C1-C20 alkyl, C2-C20 alkenyl, C3-C10 cycloalkyl aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, N(R10)2, NO2, R10O-, R11S(O)m-, R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, N3, -N(R10)2, R11OC(O)NR10-, -C(O)N(R10)2 and C1-C20 alkyl, and d) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C10 cycloalkyl; or or R2 or R3 are combined with R6 to form a ring such that is R4a, R4b, R7a and R7b are independently selected from:
a) hydrogen, b) C1-C6 alkyl unsubstituted or substituted by alkenyl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, N3, (R10)2N-C(NR10)-, R10C(O)-, -C(O)N(R10)2, -N(R10)2, or R11OC(O)NR10-, c) aryl, heterocycle, cycloalkyl, alkenyl, R10O-, R11S(O)m, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, -C(O)N(R10)2, N3, -N(R10)2, or R11OC(O)NR10-, and d) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-C10 cycloalkyl;

R5a and R5b are independently selected from:
a) hydrogen, b) substituted or unsubstituted C1-C20 alkyl, C2-C20 alkenyl, C3-C10 cycloalkyl, aryl or heterocycle group, wherein the substituent is selected from F, Cl, Br, (R10)2NC(O)-, NO2, R10O-, R11S(O)m-, R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, -CON(R10)2, N3, -N(R10)2, R11OC(O)NR10- and C1-C20 alkyl, d) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C10 cycloalkyl; or R5a and R5b are combined to form -(CH2)s- wherein one of the carbon atoms is optionally replaced by a moiety selected from: O, S(O)m, -NC(O)-, and -N(COR10)-;

R6 is independently selected from hydrogen or C1-C6 alkyl;

R8 is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R10O-, R11S(O)m-, R10C(O)NR10-, CN, NO2, R102N-C(NR10)-, R10C(O)-, -C(O)N(R10)2, N3, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R10O-, R11S(O)m-, R10C(O)NH-, CN, H2N-C(NH)-, R10C(O)-, N3, -N(R10)2, or R10OC(O)NH;

R9 is selected from:
a) hydrogen, b) alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R10O-, R11S(O)m-, R10C(O)NR10-, CN, NO2, (R10)2N-C-(NR10)-, R10C(O)-, -C(O)N(R10)2, N3.
-N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl unsubstituted or substituted by perfluoroalkyl, F, Cl, Br, R10O-, R11S(O)m-, R10C(O)NR10-, CN.

(R10)2N-C(NR10)-, R10C(O)-, -C(O)N(R10)2, N3, -N(R10)2, or R11OC(O)NR10-;
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl and aryl;
R11 is independently selected from C1-C6 alkyl and aryl;
A1 and A2 are independently selected from: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR10-, -NR10C(O)-, O, -N(R10)-, -S(O)2N(R10)-, -N(R10)S(O)2-, or S(O)m;

Q is a substituted or unsubstituted nitrogen-containing C4-C9 mono or bicyclic ring system, wherein the non-nitrogen containing ring may be a C5 to C7 saturated ring;

V is selected from:
a) hydrogen, b) heterocycle, c) aryl, d) C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and e) 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 is a heterocycle;
X, Y and Z are independently H2 or O;

m is 0, 1 or 2;
n is 0, 1, 2, 3 or 4;
p is 0, 1, 2, 3 or 4;
r is 0 to 5, provided that r is 0 when V is hydrogen;
s is 4 or 5;
t is 3, 4 or 5; and u is 0 or 1;

or a pharmaceutically acceptable salt, hydrate, crystal form or isomer thereof.

2. A compound according to Claim 1 illustrated by the formula II:

wherein:
R1a is independently selected from: hydrogen or C1-C6 alkyl;

R1b is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, R10O-, -N(R10)2 or alkenyl, c) C1-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, R10O-, or -N(R10)2;

R2 and R3 are independently selected from:
a) a side chain of a naturally occurring amino acid, b) an oxidized form of a side chain of a naturally occurring amino acid which is:
i) methionine sulfoxide, or ii) methionine sulfone, c) substituted or unsubstituted C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, NO2, R10O-, R11S(O)m-, R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3, -N(R10)2, R11OC(O)NR10- and C1-C20 alkyl, and d) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C10 cycloalkyl;
or R2 or R3 are combined with R6 to form a ring such that is R4a sand R7a independently selected from:
a) hydrogen, b) C1-C6 alkyl unsubstituted or substituted by alkenyl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, N3, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, c) aryl, heterocycle, cycloalkyl, alkenyl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3-N(R10)2, or R11OC(O)NR10-, and d) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-C10 cycloalkyl;

R4b and R7b are hydrogen;

R5a is selected from:
a) substituted or unsubstituted C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, NO2, R10O-, R11S(O)m-, R10C(O)NR10-, (R10)2NC(O)-, CN, (R10)2N-C(NR10)-, R10C(O)-, -CON(R10)2, N3, -N(R10)2, R11OC(O)NR10- and C1-C20 alkyl, and b) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C10 cycloalkyl;

R5b is selected from:
a) hydrogen, and b) C1-C3 alkyl;
R6 is independently selected from hydrogen or C1-C6 alkyl;

R8 is independently selected from:
a) hydrogen, b) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10O-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R10O, R10C(O)NR10-, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-;

R9 is selected from:
a) hydrogen, b) C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl unsubstituted or substituted by C1-C6 perfluoroalkyl, F, Cl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-;
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl and aryl;

R11 is independently selected from C1-C6 alkyl and aryl;
Q is selected from:

and A1 and A2 are independently selected from: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;

V is selected from:
a) hydrogen, b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, c) aryl, d) C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and e) C2-C20 alkenyl, and 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 is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or isoquinolinyl;

X, Y and Z are independently H2 or O;

m is 0, 1 or 2;
n is 0, 1, 2, 3 or 4;
p is 0, 1, 2, 3 or 4;

r is 0 to 5, provided that r is 0 when V is hydrogen;
t is 3, 4 or 5; and u is 0 or 1;

or the pharmaceutically acceptable salts, hydrates, crystal forms or isomers thereof.

3. A compound according to Claim 1 illustrated by the formula II:

wherein:

R1a is independently selected from: hydrogen or C1-C6 alkyl;

R1b is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, R10O, -N(R10)2 or alkenyl, c) C1-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, R10O-, or -N(R10)2;

R2 or R3 are combined with R6 to form a ring such that is ;

R4a and R7a independently selected from:
a) hydrogen, b) C1-C6 alkyl unsubstituted or substituted by alkenyl, R10O, R11S(O)m-, R10C(O)NR10-, CN, N3, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, c) aryl, heterocycle, cycloalkyl, alkenyl, R10O-, R11S(O)m, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3, -N(R10)2, or R11OC(O)NR10-, and d) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-C10 cycloalkyl;

R4b and R7b are hydrogen;

R5a is selected from:
a) substituted or unsubstituted C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, NO2, R10O-, R11S(O)m-, R10C(O)NR10-, (R10)2NC(O)-, CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3, -N(R10)2, R11OC(O)NR10- and C1-C20 alkyl, and b) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C10 cycloalkyl;

R5b is selected from:
a) hydrogen, and b) C1-C3 alkyl;
R6 is independently selected from hydrogen or C1-C6 alkyl;

R8 is independently selected from:
a) hydrogen, b) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10O-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10), R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R10O-, R10C(O)NR10-, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-;

R9 is selected from:
a) hydrogen, b) C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl unsubstituted or substituted by C1-C6 perfluoroalkyl, F, Cl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-;
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl and aryl;
R11 is independently selected from C1-C6 alkyl and aryl;
Q is selected from:

and A1 and A2 are independently selected from: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;

V is selected from:
a) hydrogen, b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, c) aryl, d) C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and e) C2-C20 alkenyl, and 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 is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or isoquinolinyl;
X, Y and Z are independently H2 or O;

m is 0, 1 or 2;
n is 0, 1, 2, 3 or 4;
p is 0, 1, 2, 3 or 4;
r is 0 to 5, provided that r is 0 when V is hydrogen;
t is 3, 4 or 5; and u is 0 or 1;

or the pharmaceutically acceptable salts, hydrates, crystal forms or isomers thereof.

4. A compound according to Claim 3 illustrated by the formula IV:

wherein:

R1a is independently selected from: hydrogen or C1-C6 alkyl;

R1b is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, R10O-, -N(R10)2 or alkenyl, c) C1-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, R10O-, or -N(R10)2;

R4a and R7a are independently selected from:
a) hydrogen, b) C1-C6 alkyl unsubstituted or substituted by alkenyl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, N3, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, c) aryl, heterocycle, cycloalkyl, alkenyl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3, -N(R10)2, or R11OC(O)NR10-, and d) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-C10 cycloalkyl;

R4b and R7b are hydrogen;
R5a is selected from:
a) substituted or unsubstituted C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl. Br, NO2, R10O-, R11S(O)m-, R10C(O)NR10-, (R10)2NC(O)- CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3, -N(R10)2, R11OC(O)NR10- and C1-C20 alkyl, and b) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C10 cycloalkyl;

R5b is selected from:
a) hydrogen, and b) C1-C3 alkyl;

R8 is independently selected from:
a) hydrogen, b) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10O-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10), R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R10O-, R10C(O)NR10-, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-;

R9 is selected from:
a) hydrogen, b) C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl unsubstituted or substituted by C1-C6 perfluoroalkyl, F, Cl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-;

R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl and aryl;
R11 is independently selected from C1-C6 alkyl and aryl;
Q is selected from:

and A1 and A2 are independently selected from: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;

V is selected from:
a) hydrogen, b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, c) aryl, d) C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and e) C2-C20 alkenyl, and 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 is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or isoquinolinyl;

Z is independently H2 or O;

m is 0, 1 or 2;

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

or the pharmaceutically acceptable salts, hydrates, crystal forms, or isomers thereof.

5. A compound according to Claim 4 illustrated by the formula V:

wherein:

R1a is independently selected from: hydrogen or C1-C6 alkyl;

R1b is independently selected from:
a) hydrogen, b) aryl, heterocycle, cycloalkyl, R10O-, -N(R10)2 or alkenyl, c) C1-C6 alkyl unsubstituted or substituted by aryl, heterocycle, cycloalkyl, alkenyl, R10O-, or -N(R10)2;

R4a and R7a are independently selected from:
a) hydrogen, b) C1-C6 alkyl unsubstituted or substituted by alkenyl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, N3, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, c) aryl, heterocycle, cycloalkyl, alkenyl, R10O-, R11S(O)m, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3, -N(R10)2, or R11OC(O)NR10-, and d) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocyclic and C3-C10 cycloalkyl;

R5a is selected from:
a) substituted or unsubstituted C1-C10 alkyl, C2-C10 alkenyl, C3-C10 cycloalkyl, aryl or heterocyclic group, wherein the substituent is selected from F, Cl, Br, NO2, R10O-, R11S(O)m-, R10C(O)NR10-, (R10)2NC(O)-, CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, N3, -N(R10)2, R11OC(O)NR10- and C1-C20 alkyl, and b) C1-C6 alkyl substituted with an unsubstituted or substituted group selected from aryl, heterocycle and C3-C10 cycloalkyl;

R5b is selected from:
a) hydrogen, and b) C1-C3 alkyl;

R6 is independently selected from hydrogen or C1-C6 alkyl;

R8 is independently selected from:
a) hydrogen, b) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10O-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R10O-, R10C(O)NR10-, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-;

R9 is selected from:
a) hydrogen, b) C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, NO2, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-, and c) C1-C6 alkyl unsubstituted or substituted by C1-C6 perfluoroalkyl, F, Cl, R10O-, R11S(O)m-, R10C(O)NR10-, CN, (R10)2N-C(NR10)-, R10C(O)-, CON(R10)2-, -N(R10)2, or R11OC(O)NR10-;
R10 is independently selected from hydrogen, C1-C6 alkyl, benzyl and aryl;
R11 is independently selected from C1-C6 alkyl and aryl;
A1 and A2 are independently selected from: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR10-, O, -N(R10)-, or S(O)m;

V is selected from:
a) hydrogen, b) heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, c) aryl, d) C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and e) C2-C20 alkenyl, and 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 is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl, indolyl, quinolinyl, or isoquinolinyl;

m is 0, 1 or 2;
n is 0, 1, 2, 3 or 4;
p is 0, 1, 2, 3 or 4;
r is 0 to 5, provided that r is 0 when V is hydrogen;
t is 3, 4 or 5; and u is 0 or 1;

or the pharmaceutically acceptable salts, hydrates, crystal forms or isomers thereof.

6. A compound which inhibits farnesyl-protein transferase which is:
N-[1-[1-(4-Cyanobenzyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-(3-chlorophenylmethyl)-amide N-[1-[1-(4-Cyanobenzyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-methyl-N-(3-chlorophenylmethyl)-amide N-[1-[1-(4-Cyanobenzyl)-1H-imidazol-5-ylacetyl]-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-(3-chlorophenylmethyl)-amide N-[1-[1-(4-Cyanobenzyl)-1H-imidazol-5-ylacetyl]-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-methyl-N-(3-chlorophenylmethyl)-amide N-[1-(3-[1H-Imidazol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[1-(3-[1H-Imidazol-4-yl]propionyl)-3(S)-ethylpyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[1-(1-(1-Farnesyl)-1H-imidazol-5-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[1-(1-(1-Geranyl)-1H-imidazol-5-ylacetyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[1-[1-(4-Methoxybenzyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide N-[1-[1-(4-Methoxybenzyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[1-[1-(2-Naphthylmethyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-(3-chlorophenylmethyl) amide N-[1-[1-(2-Naphthylmethyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide or the pharmaceutically acceptable salts, hydrates, crystal forms or isomers thereof.

7. The compound according to Claim 5 which inhibits farnesyl-protein transferase which is:
N-[1-(3-[1H-Imidazol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide N-[1-[1-(4-Cyanobenzyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-(3-chlorophenylmethyl)-amide ;

N-[1-[1-(4-Cyanobenzyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-methyl-N-(3-chlorophenylmethyl)-amide ;

or a pharmaceutically acceptable salt, hydrates, crystal forms or optical isomer thereof.

8. The compound according to Claim 5 which inhibits farnesyl-protein transferase which is:
N-[1-(3-[1H-Imidazol-4-yl]propionyl)-pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline-N-methyl-N-(3-chlorophenylmethyl) amide ;

or a pharmaceutically acceptable salt, hydrates, crystal forms or optical isomer thereof.

9. The compound according to Claim 5 which inhibits farnesyl-protein transferase which is:
N-[1-[1-(4-Cyanobenzyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-(3-chlorophenylmethyl)-amide ;

or a pharmaceutically acceptable salt, hydrates, crystal forms or optical isomer thereof.

10. The compound according to Claim 5 which inhibits farnesyl-protein transferase which is:

N-[1-[1-(4-Cyanobenzyl)-1H-imidazol-5-ylacetyl]pyrrolidin-2(S)-ylmethyl]-3(S)-ethyl-proline N-methyl-N-(3-chlorophenylmethyl)-amide ;

or a pharmaceutically acceptable salt, hydrates, crystal forms or optical isomer thereof.

11. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a compound of Claim 1.

12. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a compound of Claim 2.

13. A pharmaceutical composition comprising a pharmaceutical carrier, and dispersed therein, a therapeutically effective amount of a compound of Claim 5.

14. A method for inhibiting farnesylation of Ras protein which comprises administering to a mammal in need thereof a therapeutically effective amount of the composition of Claim 1.

15. A method for inhibiting farnesylation of Ras protein which comprises administering to a mammal in need thereof a therapeutically effective amount of the composition of Claim 2.

16. A method for inhibiting farnesylation of Ras protein which comprises administering to a mammal in need thereof a therapeutically effective amount of the composition of Claim 5.

17. A method for treating cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 1.

18. A method for treating cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 2.

19. A method for treating cancer which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 5.

20. A method for treating neurofibromin benign proliferative disorder which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 1.

21. A method for treating blindness related to retinal vascularization which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 1.

22. A method for treating infections from hepatitis delta and related viruses which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 1.

23. A method for preventing restenosis which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 1.

24. A method for treating polycystic kidney disease which comprises administering to a mammal in need thereof a therapeutically effective amount of a composition of Claim 1.