MXPA99001592A - Cycloalkyl inhibitors of protein farnesyltransferase - Google Patents

Abstract

Novel inhibitors of protein farnesyltransferase enzymes are described, as well as methods for the preparation and pharmaceutical compositions of the same, which are useful in controlling tissue proliferative diseases, including cancer, restenosis, atherosclerosis, psoriasis, and endometriosis.

Description

INHIBITORS PROTEIN CHLORINE FARNESILTRANSFERASE The present invention relates to compounds that can be used in the medical field to treat, in prophylactic form, or in any other, the proliferation of uncontrolled or abnormal tissues. Specifically, the present invention relates to compounds that inhibit the enzyme farnesyltransferase, which has been determined to activate ras protein which in turn activates cell division and are involved in cancer and restenosis.

BACKGROUND OF THE INVENTION Ras (or p21) protein has been extensively examined because mutant forms have been found in 20% of most human cancers and greater than 50% of colon and pancreatic carcinomas ( Gibbs JB, Cell, 1991; 65: 1, Cartwright T., et al., Chimica Oggi, 1992; 10:26). These mutants of ras proteins are deficient in the ability to feed back the regulation that is present in native ras and this deficiency is associated with its oncogenic action because the ability to stimulate normal cell division can not be controlled by regulatory cofactors. endogenous The recent discovery that the transformation activity of the ras mutant critically depends on post-translational modifications (Gibbs I, et al., Microbiol., Rev. 1989; 53: 171), has discovered an important aspect of ras function. identified new prospects for cancer therapy.

In addition to cancer, there are other conditions of uncontrolled cell proliferation that can be related to excessive expression and / or function of native ras proteins. Post-surgical vascular restenosis is said condition. The use of various surgical revascularization techniques such as graft deviation of the saphenous vein, endarterectomy and transluminal coronary angioplasty are often accompanied by complications due to uncontrolled growth of neointimal tissue, known as restenosis. The biochemical causes of restenosis are poorly understood and several growth factors and proto-oncogens have been implicated (Naftilan A. I, et al., Hypertension, 1989; 13: 706 and J. Clin. Invest., 83: 1419; Gibbons G. H "et al., Molec. Cell. Biol., 1993; 13: 3706). The fact that ras proteins are known to be involved in cell division processes makes them a candidate for intervention in many situations where cells divide without control. In direct analogy to the inhibition of mutant ras related to cancer, blockade of ras-dependent processes has the potential to reduce or eliminate the proliferation of inappropriate tissue associated with restenosis, particularly in those instances where the expression and / or function of Normal growth is exaggerated by factors that stimulate growth.

The function of ras depends on the modification of the proteins to associate them with the internal face of the membranes of the plasma. Unlike other proteins associated with the membrane, ras proteins lack the conventional transmembrane or hydrophobic sequences and are initially synthesized in a soluble form of cytosol. The association of the ras protein membrane is activated by a series of post-translational process steps that are signaled by a carboxyl terminal amino acid consensus sequence that is recognized by the protein farnesyltransferase (PFT). This consensus sequence consists of a cysteine residue located at four amino acids of the carboxyl terminus, followed by two lipophilic amino acids and the C-terminal residue. The sulfhydril group of the cysteine residue is alkylated by farnesylpyrophosphate in a reaction that is catalyzed by the protein farnesyltransferase. After prenylation, the three C-terminal amino acids are separated by an endoprotease and the recently exposed alphacarboxyl group of the phenylated cysteine is methylated by a methyl transferase. The enzymatic process of ras proteins that begins with farnesylation, allows the protein to associate with the cell membrane. Mutational analysis of oncogenic ras proteins indicates that these modifications are essential for transformation activity. The replacement of the cysteine residue of the consensus sequence with other amino acids gives a ras protein that is no longer farnesylated, no longer directed towards the cell membrane and lacks the ability to stimulate cell proliferation. (Hancock J.F. et al., Cell, 1989; 57: 1617, Schafer W.R., et al., Science, 1989; 245: 379, Casey P. J., Proc. Nati Acad, Sci. United States, 1989; 86: 8323).

Recently, farnesyltransferase protein (PFTs), also referred to as farnesyl protein transferases (FPTs) have been identified and a PFT of rat brain is purified until homogeneous (Reiss Y., et al., Bioch Soc. Trans., 1992; 20: 487-88). The enzyme is characterized as a heterodimer composed of an alpha subunit (49 kDa) and a beta subunit (46 Kda), both of which are necessary for catalytic activity. High levels of expression of mammalian TFP in a baculovirus system and purification of the recombinant enzyme in active form have also been achieved (Chen W. J., et al., J. Biol. Chem. 1993; 268: 9675).

In light of the above, the discovery that the function of oncogenic ras proteins is critically dependent on their post-translational process, provides a means of cancer chemotherapy through the inhibition of the enzymes they process. The identification and isolation of farnesyltransferase that catalyzes the addition of a farnesil group to ras proteins, offers a promising objective for such intervention. Ras farnesyltransferase inhibitors have been shown to have anticancer activity in several recent articles.

Ris inhibiting agents act by inhibiting farnesyltransferase, the enzyme that anchors the gene protein product to the cell membrane. The role of the ras mutation in the transduction growth signals within the cancer cells lies in the fact that the protein is in the cell membrane, so that farnesyl transferase is inhibited, the ras protein will remain in the cytosol and will be able to transmit growth signals: these factors are well known in the literature.

A peptidomimetic inhibitor of farnesyl transferase B956 and its methyl ester B1086 at 100 mg / kg have been shown to inhibit tumor growth by human bladder carcinoma EJ-1, human fibrosarcoma HT1080 and human colon carcinoma xenografts in hairless mice (Nagasu, T. et al., Cancer Res. 1995; 55: 5310-5314). In addition, the inhibition of tumor growth by B956 has been shown to be correlated with the inhibition of the post-translational process of ras in the tumor. Other ras farnesyl transferase inhibitors have been shown to specifically avoid ras processing and membrane localization, and are effective in reversing the transformed phenotype of cells containing the mutant ras (Sepp-Lorenzino L., et al., Res Cancer). ., 1995; 55: 5302-5309).

In another report (Sun J., et al., Cancer Res., 1955; 55: 4243-4247), an inhibitor of ras farnesyl transferase FT1276 has been shown to selectively block tumor growth in hairless mice of a human lung carcinoma. with K -ras mutation and p53 elimination. In yet another report, daily administration of a farnesyl transferase inhibitor ras L-744, 832 caused tumor regression of mammary and salivary carcinomas in ras transgenic mice (Khol et al., Nature Med., 1995; 1 (8): 792- 748). Thus, farnesyl transferase inhibitors have benefits in certain forms of ras. particularly those that depend on oncogenic for their growth. However, it is well known that human cancer often manifests when several mutations occur in important genes, one or more of which may be responsible for controlled growth and metastasis. A single mutation may not be enough to sustain growth and only after two or three mutations occur can tumors develop and grow. It is therefore difficult to determine which of these mutations may be mainly those that lead to growth in a particular type of cancer. Thus, ras farnesyl transferase inhibitors may have therapeutic utility in tumors not only dependent on oncogenic ras forms for their growth. For example, several ras famesyl transferase inhibitors have been shown to have antiproliferative effects in vivo against tumor lines, either wild-type or ras mutant (Sepp-Lorenzino, supra.). In addition, there are several related to ras that are prepiled. Proteins such as R-ras2 / TC21 are ras-related proteins that are prenylated in vivo by famesyl transferase and geranylgeranyl transferase I (Carboni, et al., Oncogene, 1995; 10: 1905-1913). Therefore ras famesyl transferase inhibitors could also block the reward of the above proteins and thus would be useful in the inhibition of tumor growth driven by other oncogens.

With respect to restenosis and vascular proliferative diseases, inhibition of cellular ras has been shown to prevent smooth muscle proliferation after vascular injury in vivo (Indolfi C, et al., Nature Med., 1995; 1 (6). ): 541-545). This report definitely supports a role for famesyl transferase inhibitors in this disease, showing inhibition of accumulation and proliferation of vascular smooth muscle.

SUMMARY OF THE INVENTION The present invention supplies the compounds having Formula (I). where: RQ is - C6 aiquil? X is O or 1; each R is independently hydrogen or Ci-C6 alkyl, A is - CORa, - CO2Ra \ - CSR ', O S II II - CSR \ - CNRaRa, \ - C (S) ORa \ - C (S) NHRa \ - SO2Ra, or - CONRaRa, Ra, Ra 'and Ra "are independently Ci-C6 alkyl, - (CR RH) m-cycloalkyl, - (CR 14 R 14 m-aryl, or - (CR 14 R 1 m -heteroaryl; Each m is independently 0 to 3, R1, R2, and R4 are independently hydrogen or Ci-C6 alkyl.

(\ RJ is - (CH2) m CH (CH2) C, -C? Alkyl, V J C2-C (, alkenyl.

(CRHR! 4) m - naphthyl, - (CH2) vCO2R, (CH2) tNRl4R14, (CH2) V - O - C, - Cr, alkyl, - (CH2? - OH, (CH2X - morpholine? - (CH2? NN R14, C (CH2) V (NR14 \ / \ / - (CR! 4ER54) m - (phenyl substituted with Rb), or - (CR! R! 4) m - (heteroaryl substituted with Rb) t is 2 to 6, v is 1 to 6 Rb is - O - phenyl, - O - benzyl, halogen, G - C (, alkyl, hydrogen, - O Ci - C6 alkyl, - NH2, - NHRa, - NR'R ", OOO II II II - CCi - Cr, alkyl, - C - aryl, - OH, - CF3, - NO2, - COH, O O II II - COC, - alkyl, - CN, - OPO3H2, - CH2PO3H2, - CO aryl, O O O O - N3, - CF2CF3, - SO2Ra, - SO2NRaRa - CHO, --OCOCH3, --O (CH2) m - heler? Aiil, --O (CH2) m - ail, - (CH2) m - cycloalkyl, - ( CH2) m-heteroaryl, or - CH = CHC6C5, and is 2 or 3; R5 is (feni I substituted with Rd, Rh and R1) (phenyl substituted with Re, Rh and R1) each ti is iiidepeiidienteineiite 2, 3 or 4, R1, Re and Rh are independently hydrogen, halogen, - OCi - Ce - alkyl, C? - Cr, alkyl, - CN, - OPO3H2; ? ? II II - CH2PO3H2, - O - phenyl, - O - benzyl, - CNH2, - CNHRa O O ¡I ¡¡ CNRaRa \ - NHCRa, - O (CH2) vNRaRa \ - NH2, NHR \ O O II II - NRaRa \ - CC, - C6 - alkyl, - C - aryl, - OH, - CF3, - NO2, OO O. II II II - COH, - COCy - C6 alkyl, - CO ail, - N3, - CF2CF3, - SO2Ra, - SO2NRaRa - CHO, or - OCOCH3; and R c and R d are independently C] -C 6 alkyl, - (CH 2 m -cycloaicyl or hydrogen, and the pharmaceutically acceptable salts, esters, amides and pr? di? gas thereof.

In an ejeinμhti μiedilecl? of the in? ues of the F? imuki I. R1 is hydrogen, R2 is hydrogen, R is hydrogen, R! 1 is hydrogen or methyl, and A is In? Tr? favorite copy of l? s c? mpuest? s of the F? imula 1.

R3 is (CH2) mCH CH2 ?, Ci C6 alkyl, C2 - O alkenyl, - (Cll2) m - (phenyl substituted with Rh) O - (C? 2) m - (heteroaryl substituted with Rb).

R1 is hydrogen, R2 is hydrogen, R4 is hydrogen and R14 is hydrogen or methyl.

In ll? exemplai piedilecl? of l? s c? mpuesl? s of the F? imula I. R5 is - Í - - - - phenyl, substituted with fluoro, chloro, bromo or NH2 at the site - 2), or The ciphoses that have the Formula II are also supplied.

Where Ci - Cr, alkyl? R6 is -O-benzii, -OCH-phenyl, -NH-benzyl-N (C? -C6 alkyl) -benzyl, or -CH2-phenyl; R8 is hydrogen, halogen, Ct-Ce alkyl, -O-benzyl, -OCH2-p idyl, -OC, -C6 alkyl ?. - CF3, - OH, or feml, R10 and R13 are independently hydrogen or C? - Ce alkyl; each n is independently 2, 3, or 4, R14 is hydrogen or methyl, R ', R! T and R! are independently hydrogen, halogen, - NH2, - NHRa, - OCj -? alkyl,? - Ci - Ce alkyl * and the salts, steels, amides and pi? Diogas faimaceulicamenle acceptable of themselves The compounds having the Formula III were also supplied. where: each n is 2, 3 or 4, X is NH, O, or - NCH3; R15 is -O-benzyl, -FC3, hydrogen, halogen, -OH, -phenyl, -CI-C6 alkyl, -OCH2-pyridyl,? - Od - Ce alkyl? and pharmaceutically acceptable salts, esters, amides and pi? di? gas thereof.

A c? Inpuest is also supplied? that has the F? iula IV where. X is H, O, or - NCH3; R 4 is hydrogen or C 2 - (, alkyl, R 4 is hydrogen or C 2 - Ce alkyl, C 2 - Ce alkenyl, - ((CH 2)? NR 1'lRM, - (CH 2) V - O - Cj - C 6 - alkyl? - (CH2? - OH, (CH2-14, - (CII2) vn is 2, 3 or 4, m is 0 to 3, L is 2 to 6, and v is 1 to 6.

In particular, the invention provides a pharmaceutically acceptable composition comprising a compound of Formula I, II, II, or IV.

A rnét? D is also supplied? To treat ? To prevent the restenosis, the method comprises the admission to a patient who has stenosis or risk of tenei stenosis, of a therapeutically effective quality of a compound of Formula 1, 11, III or IV.

A method for treating cancer is also provided, the method comprising administering to a patient having cancer a therapeutically effective amount of a compound of Formula I, p, III, or IV.

In a more preferred specimen, cancer is lung cancer, colon cancer, breast cancer, pancreatic cancer, thyroid cancer or bladder cancer.

In a more preferred example, the compounds of Formulas I, p, ni or IV are: Acid (S) - [1 - ((4-benzyloxy-benzyl) - { [(1-phenyl-cyclobutylmethyl) -carbamoyl) ] - methyl.}. - carbamoyl) -2- (1H-imidazol-4-yl) -ethyl] -carbamic acid benzyl ester; (S) - [1 - ((4-Benzyloxy-benzyl) -. {[[(1-phenyl-cyclopropylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - [1 - ((4-Benzyloxy-benzyl) -. {[[(1-phenyl-cyclopentylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - [1 - ((4-Phenyl-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} - carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - [1 - ((4-methoxy-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - [1 - ((4-Methyl-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} - carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - N - (4-Benzyloxy-benzyl) -2- (3-benzyl-ureido) -3- (1 H -imidazol-4-yl) -N- acid. { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} -propionamide; (S) -2- (3-Benzyl-3-methyl-ureido) -N - (4-benzyloxy-benzyl) -3- (1H-imidazol-4-yl) -N- acid. { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} -propionamide; (S) - [1 - ((4-Benzyloxy-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - thiocarbamic S benzyl ester; (S) - (2 - (1H-imidazol-4-yl) -1- {. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] - [4 - (pyridine-2) acid - il-methoxy) -benzyl] -carbamoyl.} - ethylcarbamic benzyl ester; (S) - [1 - ((Cyclohexyl-methyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] methyl} - carbamoyl) -2 - (1H-imidazol-4-yl) -ethyl acid ] - carbamic benzyl ester; (S) - (1 - ((isobutyl) - { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl.}. Carbamoyl) -2 - (1H-imidazol-4-yl) -ethyl] - carbamic S-benzyl ester; (S) -2- (3-Benzyl-3-methyl-ureido) -3- (1H-imidazol-4-yl) - N - (4-methyl-benzyl) -N- acid. { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} -propionamide; (S) - [1 - [(4-Benzyloxy-benzyl) - ( { [1 - (2,6-dichloro-phenyl) -cyclobutylmethyl] -carbamoyl.} - methyl) -carbamoyl] -2 - ( 1H-imidazol-4-yl) -ethyl] -carbamic benzyl ester; [(S) -trans] - [1 - (But-2-enyl - {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} - carbamoyl) -2 - (3H-imidazole-4) acid - il) - ethyl] - carbamic benzyl ester; (S) - (2 - (3H-imidazol-4-yl) -1 - ( { [1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} - propylcarbamoyl) -ethyl] -carbamic acid benzyl ester ester; Benzyl N - ((SS) - 1 - (1 H - 4 - imidazolylmethyl) -2-oxo-2 - (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) [(IR) -1-phenylethyl] aminoethyl) carbamate; (S) - [1 - ((1,1-dimethyl-2-phenyl-ethyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2- (3H) acid imidazol-4-yl) -ethyl] -carbamic acid benzyl ester; Benzyl N - ((SS) - 1 - (1 H - 4 - imidazolylmethyl) -2-oxo-2 - (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) [(SS) -1-phenylethyl] aminoethyl) carbamate; Benzyl N - [(SS) - 2 - [(2-hydroxyethyl) (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) amino] -1- (1H-4-imidazolylmethyl) -2-oxoethyl] carbamate; 3 - [[(2S) -2 - [(benzyloxy) carbonyl] amino-3 - (lH-4-imidazolyl) propanoyl] (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] propanic; Methyl-3 - [[(2S) -2 - [(benzyloxy) carbonyl] amino-3 - (1H-4-imidazolyl) propanoyl] (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino propanoate; Benzyl N - [(SS) - 2 - [(2-aminoxyethyl) (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) amino] -1 - (lH-4-imidazolylmethyl) -2-oxoethyl] carbamate; Benzyl N - (lS) - (lH-4-imidazolylmethyl) -2 - [[2- (methylamino) ethyl] (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] -2-oxoethylcarbamate; Benzyl N - (1 S) - (1 H -4-imidazolylmethyl) -2 - [(2 - (methoxyethyl) (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] -2-oxoethylcarbamate; Benzyl N - (lS) - (lH-4-imidazolylmethyl) -2 - [(2 - (methyl-2-phenylpropyl) (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) amino] -2- oxoethylcarbamate; and 1 - . 1-Phenylethyl N - [(SS) -2 - [[4 - (benzyloxy) -1,5-cyclohexadienyl] methyl (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] -1 - (1H-4-imidazolylmethyl) -2-oxoethyl] carbamate.

DESCRIPTION OF ALLAND OF THE INVENTION The present compound provides an e? Rnpuest? which has the formula I. where: RQ is C1 ~ C6 a'lliuu0 I D x is 0 or i, each R "is independently hydrogen or G-Ce alkyl, A is - CORa, - CO2Ra \ - CSRa, O !! 0 - CSR ', - CNRaRa ", - C (S) ORa \ - C (S) NHR", - SO2Ra, or - CONR'R' "; Ra, Ra and Ra 'are independently Ct - (, alkyl, - (CR14R14) m-cycloalkio, (CRR) m-aryl, or - (CRRH) m-heteroaryl; Each /// is independently 0 to 3, R !, R \ and R4 are independently hydrogen or alkyl G - C6; i \ RJ is - (CH7) m CII (CU,! C, - Ce alkyl. \ i C2 - ß alkenyl, - (CRl4R14Xn - naphthyl, - (CH2) vCO2R14" - (CH2XNR14R14, - (CH2) V - O - Ct - C6 alkyl, - (CH2? - OH, - (CH2? - morpholino - (CH2) t NNR 1144, r C / (rCuH2) V / (x Nt r R »14 - (CR14ER14) m - (phenyl substituted with Rb), or - (CR14R14) m - (heteroaryl substituted with Rb); t is 2 to 6; v is 1 to 6; Rb is -O-phenyl, -O-benzyl, halogen, Ct-C6 alkyl, hydrogen, - O Ci - C6 alkyl, - NH2, - NHRa, - NRaRa, O O O O II II II II - CCi - C6 alkyl, - C - aryl, - CNH2, - CNHRa, - CNRaRa, O O II II NHCRa, -O (CH2) and NRaRa, - OH, - CF3, - NO2, - COH, O O - COCt - Ce alkyl, - CN, - OPO3H2, - CH2PO3H2, - CO aryl, - N3, - CF2CF3, - SO2Ra, - SO2NRaRa ', - CHO, - O (CH2) m - aryl, - (CH2) m - cycloalkyl, - (CH2) m - aryl, - (CH2) m cycloalkyl, - (CH2) m heteroaryl, CH = CHCeCs, - OCOCH3, or - O (CH2) m heteroaryl and is 2 or 3; R5 is (phenyl substituted with Re, Rh and K ') (phenyl substituted with Rg., Rh and R. ') each // is independently 2, 3 or 4, R ', Re and Rh are independently hydrogen, halogen, - OC, - C6 - alkyl ?, Q - C6 alkyl? - CN, - OPO3H2; O O II II - CH2PO3H2, - O - pheny, - O - benzyl, - CNH2, - CNHRa 0 O || CNR'R ", - NHCR", - O (CH2) and NR "R", - NH2, NHRa, O O 'II II - NRaRa \ - CCi - Ce - alkyl, - C - aryl, - OH, - CF3, - NO2, O ü O II II I - COH, - COC, - Ce alkyl, - CO aryl, - N ,, - CFjCF ,, - SO2Ra, - SO2NRaRa \ - CHO, or - OCOCH », and Rc and R are independently G - Cβ alkyl, - (CH2) m - cycloalkyl or hydiogen ?, and the faimaceulicamenle acceptable salts, esters, amides and piodiogas thereof .

The competitors that have the Formula II were also supplied. where C, - Cr, alkyl? R6 is -O-benzyl, -OCH-phenyl, -NH-benzyl-N (G-C6 alkyl?) -benzyl,? - SCH2 - phenyl, R8 is hydrogen, halogen, Ct - C alkyl, - O - benzyl, - OCH2 - μiiidyl, - OG - C6 alkyl ?. - CF3, - OH, or phenyl, R 10 and R 13 are independently hydrogen or C, -Cß alkyl; each n is independently 2, 3,? 4, R14 is hydrogen or methyl; R Rk and R! are independently hydrogen, halogen, - NH2, - NHRa, - OG - Ce alkyl,? - G - Cr, alkyl? and the pharmaceutically acceptable salts, esters, amides and prodrug of the same substances.

The following are also supplied with Formula III: where: Each // is 2, 3 or 4, X is NH, O, or - NCH3, R15 is - O - benzyl, - CF3, hydrogen, halogen, - OH, - phenyl, - G - C6 alkyl, - OCH2 - μiiidil, or - OCi - Cr, alkyl? and the pharmaceutically acceptable salts, esters, amides and prodrug of the same.

A compound having the Fíiula IV was also supplied. where. X is H, O, or - NCH3, Rra is G - C6 alkyl, C2 - C6 alkenyl, - (CH ^ NR ^ R14, - (CH2) V - O - C, - C6 - alkyl, - (CH2? - OH, - (CH2? - (iH4 '- (CH2X - each R is hydrogen or G-C6 alkyl, n is 2, 3? 4, m is 0 to 3, t is 2 to 6, and v is 1 to 6.

The term in? "rent?" does it mean a linear hydrocarbon? branched? which has from 1 to 6 caybon atoms and includes, for example, methyl, elil, n-piopil, isopropyl, n-butyl, sec -butyl, isylbutyl, tert -butyl, n -pentyl, p -ylxyl and similar.

The tea min? "Cycle? Rent?" means a saturated carbide hydrocarbon chain which has from 3 to 7 carbon atoms, for example, cyclopiopyl, cyclobutyl, cyclic pentyl, cyclohexyl, adamantyl and the like.

F. l téimin? "aryl" means an aromatic chain which is a phenyl, 5-luoyil, 1-yiafyl,? 2 - nafl.il, unsubstituted? substiluido p? i 1 to 3 subslituyenles, selected from alkyl, O-alkyl? and S-alkyl ?, OH, SH, F, Cl, Bi, I, CF3, NO2, H, NHCH3, N (CH3) 2, NHCO-alkyl, -OCH2CeH5, (CH2) m CO2-alkyl, (CH2) m SO3H, (CH2) mPO3H2, (CH2) m PO3 (alkyl) 2, (CH2) m SO2NH2, and (CH2) m SO2NH-alkyl, where the alkyl is defined above and M = 0, 1, 2, or 3.

The term "heteroaryl" means a heteroaromatic chain which is a group 2-or 3-thienyl, 2- or 3-furanyl, 2- or 3-pyrrolyl, 2- or 3-or 4-pyridyl, imidazolyl, 2 -, 3 -, 4 -, 5 -, 6 -. or 7-indolyl, unsubstituted or substituted by 1 or 2 substituents of the group of substituents described above for aryl.

The symbol "-" means a link.

The term "patient" means all animals including humans. Examples of patients include humans, cows, dogs, cats, goats, sheep and pigs.

A "therapeutically effective amount" in an amount of a compound of the present invention that when administered to a patient, improves a symptom of restenosis or cancer or prevents restenosis. A therapeutically effective amount of a compound of the present invention can be readily determined by one skilled in the art by administering an amount of a compound to a patient and observing the result. In addition, those skilled in the art are familiar with identifying patients who have cancer or restenosis or who are at risk of having restenosis.

The term "cancer" includes, but is not limited to the following cancers: breast; ovary; cervix; prostate; testicle; esophagus; glioblastoma; neuroblastoma; stomach; skin; ceratoacanthoma; lung, squamous cell carcinoma; large cell carcinoma; adenocarcinoma; bone; colon; adenocarcinoma; adenoma; pancreas, adenocarcinoma; thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma; seminoma; melanoma; sarcoma; bladder carcinoma; carcinoma of the liver and bile ducts; kidney carcinoma; myeloid disorders; Limfoid disorders Hodgkins, hair cells; oral cavity and pharynx (oral), lips, tongue, mouth, pharynx; small intestine; colon - rectum, large intestine, rectum; brain and central nervous system; and leukemia.

The term "pharmaceutically acceptable salts, esters, amides and prodrugs" as used herein, refers to those carboxylate salts, amino acid addition salts, esters, amides and prodrugs of the compounds of the present invention which are, within of the scope of what sounds like medical judgment, appropriate to be used in contact with the tissues of patients without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable benefit / risk ratio and effective for their intended use, as well as zwitterionic forms, where possible, of the compounds of the invention. The term "salts" refers to the non-toxic, inorganic and organic acid addition salts of the compounds of the present invention. These salts can be prepared in situ during the isolation and final purification of the compounds or by reacting separately the purified compounds in their free base form with an appropriate organic or inorganic acid and isolating the salt thus formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, mesylate salts of nephrylate, glucoheptonate, lactobionate and lauryl sulphonate and the like. These may include cations based on alkali metals and alkali metals, such as sodium, lithium, potassium, calcium, magnesium and the like, as well as non-toxic ammonium, quaternary ammonium and amine cations including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine , dimethylamine, trimethylamine, triethylamine, ethylamine and the like, (see, for example, SM Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977; 66: 1-19 which is incorporated herein) as reference).

Examples of pharmaceutically acceptable non-toxic esters of the compounds of this invention include G-C alkyl esters where the alkyl group is a straight or branched chain. Acceptable esters also include G-C7 cycloalkyl esters, as well as arylalkyl esters such as, but not limited to, benzyl. The G - C esters are favorite. The esters of the compounds of the present invention can be prepared according to conventional methods.

Examples of pharmaceutically acceptable non-toxic amides of the compounds of this invention include amides derived from ammonia, primary amines G-Cß alkyl and secondary amines Ct-C6 dialkyl, where the alkyl groups are straight or branched chains. In the case of secondary amines, the amine may also be in the form of a 5- or 6-membered heterocycle containing a nitrogen atom. The amides derived from ammonia, primary amines Ct-C6 alkyl and secondary amines G-C6 alkyl are preferred. The amides of the compounds of the invention can be prepared according to conventional methods.

The term "prodrug" refers to compounds that are rapidly transformed in vivo to produce the original compound of the above formulas, for example, by hydrolysis in the blood. A full discussion is given in T. Higuchi and V. Stella, "Pro -drugs as Novel Delivery Systems," vol. 14 of the symposium series A. C. S., and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche. American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference.

The compounds of the present invention can be administered to patients alone or as part of a composition containing other components such as excipients, diluents and carriers, all of which are well known in the art. The compositions can be administered to humans and animals, either oral, rectal, parenteral (intravenous, intramuscular or subcutaneous), intracisternal, intravaginal, intraperitoneal, intravesicular, locally (powders, ointments or drops) or as a buccal or nasal sprinkler.

Compositions suitable for parenteral injection may comprise physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution within sterile injectable solutions or dispersions. Examples of suitable aqueous or non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol and the like), appropriate mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by using a coating such as lecithin, by maintaining the required particle size in the case of dispersions and by the use of surfactants.

These compositions may also contain adjuvants such as preservatives, humectants, emulsifiers and dispersants. The prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid and the like. It may also be desirable to include isotonic agents, for example sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be obtained by the use of delayed absorption agents, for example, aluminum monostearate and gelatin.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In said solid dosage forms, the active compound is mixed with at least one regularly inert excipient (or carrier) such as sodium citrate and dicalcium phosphate or (a) fillers or extenders, such as, for example, starches, lactose, sucrose, glucose , mannitol and silicic acid (b) binders, such as, for example, carboxymethyl cellulose, alinates, gelatin, polyvinyl pyrrolidone, sucrose and acacia; disintegrating agents, such as, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and sodium carbonate; (e) solution retarders, such as paraffin; (f) absorption accelerators, such as, for example, quaternary ammonium compounds; (g) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (h) absorbers, such as, for example, kaolin or bentonite; and (i) lubricants, such as, for example, talc, calcium stearate, magnesium stearate, polyethylene glycols, sodium lauryl sulfate or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms also comprise regulatory agents.

Solid compositions of a similar type can also be used as fillers in soft or hard gelatin capsules using such excipients as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with layers and shells, as enteric layers and others well known in the art. These may contain opacifying agents and may also be of said composition in that they release the active compound or compounds in a certain part of the digestive tract in a delayed manner. Examples of inserted compositions that can be used are polymeric substances and waxes. The active compounds may also be in micro-encapsulated forms, if appropriate, with one or more of the aforementioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as, for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate , benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3. butylene glycol, dimethylformamide, oils, in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, glycerol, tetrahydrofurfuryl alcohol, Cremophor EL (a derivative of castor oil and ethylene oxide) obtained in Sigma Chemical Co., St. Louis, Missouri), polyethylene glycols and esters of sorbitan fatty acids or mixtures of these substances and the like. - - - - Apart from said inert diluents, the composition may also include adjuvants, such as wetting agents, emulsifiers and suspending, sweetening, flavoring and perfuming agents.

The suspensions, in addition to the active compounds, may contain suspending agents, such as, for example, ethoxylated isostearyl alcohols, polyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances and Similar.

Compositions for rectal administration are preferably suppositories which can be prepared by mixing the compounds of the present invention with non-irritating excipients or carriers such as, for example, cocoa butter, polyethylene glycol or suppository wax, which are solid at temperatures ordinary, but liquid at body temperature and, therefore, melt in the rectum or vaginal cavity and the active compound is released.

Dosage forms for topical administration of a compound of this invention include ointments, powders, sprays and inhalants. The active compound is mixed under sterile conditions with a physiologically acceptable carrier and any condom, regulator or propellant that is required. Ophthalmic formulations, ointments for eyes, powders and solutions are also contemplated as being within the scope of this invention.

The compounds of the present invention can be administered to a patient at dosage levels in the range of 0.1 to 2,000 mg per day. For a normal human adult who has a body weight of about 70 Kg, a dosage in the range of 0.01 to 100 mg per kilogram of body weight is preferable. The specific dosage used, however, may vary. For example, the dosage may depend on several factors including the requirements of the patients, the severity of the condition being treated and the pharmacological activity of the compound used. The determination of optimal doses for a particular patient is well known to those skilled in the art.

The compounds of the present invention can exist in different stereoisomeric forms by virtue of the presence of asymmetric centers in the compounds. It is contemplated that all stereoisomeric forms of the compounds, as well as mixtures thereof, including racemic mixtures, form part of this invention.

In addition, the compounds of the present invention may exist in insolvated forms, as well as solvated forms with pharmaceutically acceptable solvents, such as water, ethanol, and the like. In general, Solvated forms are considered equivalent to the insolvated forms for the purposes of present invention.

It is intended that the examples presented below illustrate the particular instances of the invention, and are not intended to limit the scope of the specification of the claims in any way.

Scheme 1 shows the general method by which the compounds of the present invention can be prepared.

SCHEME 1 Preparation of Example 1 SCHEME 2 Piepai ación del Ejempl? 10 50 ', nqnooiis HOAc In step 4 of Scheme 1 the alternative condensing agents PyBOP; dicyclohexylcarbodiimide (DCC / l-hydroxy-benzotriazole (HOBt), O-benzotriazol-1 -yl-N, N, N \ N \ - tetra-methyluronium hexafluorophosphate (HBTU), and O - (7-azabenzotriazol-1-yl) - N, N, N \ N '- tetramethyl uronium hexafluorophosphate (HATU) / 1-hydroxy-7-azabenzotriazole (HOAt), can also be used In step 5, you can also use 80% acetic acid / water 90 ° C for 30 minutes.

Abbreviations: DCM Dichloromethane Na (Oac) 3BH triacetoxy sodium borohydride PyBOP benzotriazol-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate DIEA Diisopropyl ethylamine. THF Tetrahydrofurano BOP Benzotriazol-1-yl-oxy-tris hexafluorophosphate (dimethylamino) - phosphonium TFA Trifluoroacetic acid EA Ethyl acetate EXAMPLE 1 (S) - [- ((4-Benzyloxy-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazole-4- il) - ethyl] - carbamic benzyl ester, Step 1: [(4-benzyloxy-benzyl) -amino] -acetic methyl ester To a suspension of glycine methyl ester hydrochloride (1.26 g, 10 mmol) and 4-benzyloxy-benzaldehyde (2.12 g, 10 mmol) in DCM (50 mL), under nitrogen at 0 ° C, is added Na (Oac) 3BH (3.81 g, 15 mmol). The suspension is allowed to warm to room temperature and stirred for 4 hours. Then, the suspension is poured into saturated aqueous NaHCO3, and the layers are separated. The aqueous layer is extracted with DCM (4 x 25 mL).

The combined organic extracts are dried over MgSO and concentrated. Lightning chromatography (EA) produces 1.15 g (40.4%) of the main compound as a white solid; melting point: 57 ° - 58 ° C.

Analysis calculated for G7H19NO3: C, 71.56; H, 6.71; N, 4.91. Found: C, 71.45; H, 6.99; N, 4.92.

Mass Spectrum (MS) - (Chemical ionization (Cl): Calculated for M + l: 286, it was found: 286. The NMR spectrum is consistent with the structure.

Step 2: Acid (S) -1- (4-benzyloxy-benzyl) - [2-benzyloxycarbonylamino-3 - (1 - trityl-1H-imidazol-4-yl) -propionyl] -amino} - Acetic methyl ester. To a solution of (S) -2-benzyloxycarbonyl-amino-3 - (1-trityl-1H) - imidazol-4-yl) -propionic [Cbz-His (Trt)] (Hudspeth JP Kaltenbronn JS, Repine JT, Roark WH, Stier MA Renin Inhibitors III, U.S. Patent No. 4,735,933, 1988) (5.85 g, 11 mmoles) and PyBOP (5.72 g, 1 mmol) in DCM (100 mL) at 0 ° C. Methyl ester from step 1 above is added (2.85 g, 10 mmol) followed by DIEA (3.05 mL, 17.5 mmol). . The solution is heated to room temperature and stirred for 5 hours under nitrogen. The solution is poured into saturated aqueous NaHCO3, and the layers are separated. The aqueous layer is extracted with DCM (4 x 50 mL). The combined organic layers are dried over MgSO and concentrated. Lightning chromatography (10% methanol [MeOH] in DCM) gives 4.48 g (55.4%) of the main compound as a white foam.

Analysis calculated for C50H4 N4O3O.5 H2O C, 7433; H, 5.86; N, 6.93. Found: C, 74.00; H, 5.75; N, 6.79. Electrospray (ES): Calculated for M + 1: 799.3; It was found: 799.3.

The NMR spectrum was consistent with the structure.

Step 3: Acid (S) -. { (4-benzyloxy-benzyl) - [2-benzyloxycarbonylamino-3 - (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - acetic To a solution of the methyl ester from step 2 above (0.799, 1 mmol) in THF (10 mL) is added H2O (3.3 mL) to give a solution. The solution is cooled to 0 ° C. and treated with LiOH 2 O (0.050 g, 1.2 mmol). The solution is heated to room temperature and stirred for 4 hours. Another batch of LiOH 2 O (0.050 g, 1.2 mmol) is added and the suspension is concentrated, diluted with H 2 O and pH = 2 (paper) is adjusted with 1 M HCl. The suspension is extracted with EA (4 x 20 mL). The organic extracts are dried over MgSO4 and concentrated. Flash chromatography (10% MeOH in DCM) gives 0.706 g (89.0%) of the parent compound as a foam.

Analysis calculated for C4 H44N O6-0.5 H2O C, 74.13; H, 5.71; N, 7.05. It was found: C, 74.22; H, 5.70; N, 6.81. MS - ES: Calculated for M + l: 785.3; Found: 785.2. The NMR and ir spectra were consistent with the structure.

Step 4: Acid (S) - [1 - ((4-benzyloxy-benzyl) - ([(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl.} - carbamoyl) -2- (1-trityl-1H- imidazol-4-yl) -ethyl] -carbamic benzyl ester To a suspension of the acid from step 3 above (0.500 g, 0.637 mmol), BOP (0.310 g, 0.70 mmol) and C - (1-phenyl-cyclobutyl) hydrochloride ) - methylamine (Bridges A.

I, Hamilton H. W., Moos W. H., Szotek D. L., "N-Substituted Adenosines". U.S. Patent No. 4,755,594; 1988) (0.139 g, 0.701 mmol) in DCM (10 mL) a 0 ° C, DIEA (0.29 mL, 1.75 mmol) is added. The resulting solution is stirred at room temperature overnight. The solution is poured into saturated NaHCO3 and the layers are separated. The aqueous layer is extracted with DCM (3 x 50 mL). The combined organic layers are washed with H2O, then twice with 0.5 M HCl, then once with Saturated aqueous NaHCO3, dried over MgSO4 and concentrated. Lightning chromatography (10% MeOH in DCM) gives 0.49 g of the main compound as a white foam which is not characterized and used directly in the next reaction.

Step 5: (S) - [1 - ((4-Benzyloxy-benzyl) - { [(1-phenyl-cyclobutylmethi -carbamoyl] -methyl] -carbamoyl) -2- (1H-imidazole-4) acid - il) - ethyl] - carbamic benzyl ester To a solution of trityl from step 4 above (0.49 g) in DCM is added TFA (10 mL) The solution is stirred at room temperature for 2 hours and then concentrated The residue is dissolved in DCM and washed with saturated aqueous NaHCO3 The aqueous layer is extracted with DCM (2 x 50 mL) The combined organic extracts are dried over MgSO 4 and concentrate. Lightning chromatography (10% MeOH in DCM) gives 0.22 g (50% yield over two steps) of the main compound as a white foam.

Analysis calculated for QtHßNsOs-O.S H2O C, 70.87; H, 6.38; N, 10.08. Found: C, 70.81; H, 6.46; N, 9.91. MS - Chemical Ionization of Atmospheric Pressure (APCI): Calculated for M + l: 686.3; It was found: 686.3. The NMR and ir spectra were consistent with the structure.

EXAMPLE 2 Acid (S) - [1 - ((4-benzyloxy-benzyl) - { [(1-phenyl-cyclopropylmethyl) -carbamoyl] -methyl I-carbamoyl) -2 - (1H-imidazol-4-yl) ) - ethyl] - carbamic benzyl ester. According to Example 1, Steps 4 and 5, by substituting C - (1-phenyl-cyclopropyl) -methylamine hydrochloride (U.S. Patent No. 4,755,594; 1988) for C - (1-phenyl-cyclobutyl) hydrochloride - methylamine, 0.11 g (26% yield over two steps) of the main compound is obtained as a white foam. Analysis calculated for GOHHNÍOS'O.S H2O C, 70.57; H, 6.22; N, 10.29. Found: C, 70.43; H, 5.98; N, 10.25.

MS - APCI: Calculated for M + l: 672.3; it was found: 672.3. The NMR and ir spectra were consistent with the structure.

EXAMPLE 3 Acid (S) - (2 - (1H-imidazol-4-yl) -1 - (([(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl 1 - [4 - (pyridin-2-ylmethoxy ) - benzyl] -carbamoyl.} - ethylcarbamic acid benzyl ester Step 1: [(4 - (pyridin-2-ylmethoxy) -benzyl) -amino] -acetic acid methyl ester According to Example 1, Step 1, by substituting 4- (2-pyridinyl-methoxy) benzaldehyde (J. Het, Chem., 1988; 25: 129) for 4 -4-benzyloxy-benzaldehyde, the main compound is obtained as an oil (36.3% yield). MS-Cl: calculated for M + l: 287; Found: 287.

Step 2: Acid (S) -. { (4 - (pyridin-2-ylmethoxy) -benzyl) - [2-benzyloxy -carbonyl-amino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - Acetic methyl ester. According to Example 1, Step 2, when replacing the compound of Step 1 above with [(4-benzyloxy-benzyl) -amino] -acetic methyl ester, the main compound is obtained as a white foam (90.1% yield). MS - APCI: Calculated for M + l: 800.3; It was found: 800.3.

Step 3: Acid (S) -. { (4 - (pyridin-2-ylmethoxy) -benzyl) - [2-benzyloxy -carbonyl-amino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - acetic According to Example 1, Step 3, by replacing the methyl ester of Step 2 above with (S) - acid. { (4-benzyloxy-benzyl) - [2-benzyloxy-carbonyl-amino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - Acetic methyl ester, the main compound is obtained as a white foam (87.5% production). MS - APCI: Calculated for M + 1: 786.3; was found: 786.3.

Step 4: (S) - [1 - (((4-pyridin-2-ylmethoxy) -benzyl) - [1 - [(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methylcarbamoyl) -2- ( 1-trityl-1H-imidazol-4-yl) -ethyl] -carbamic acid benzyl ester. According to Example 1, Step 4, by replacing the acid from Step 3 above with acid (S) -. { (4-Benzyloxy-benzyl) - [2-benzyloxy-carbonyl-amino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - acetic, the main compound is obtained as a clear oil (100% production). MS - APCI: Calculated for M + l: 929.4; it was found: 929.2.

Step 5: fS) - (2 - (1H-imidazol-4-yl) -1 - (([1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl- [4 - (pyridin-2-ylmethoxy) - benzyl] -carbamoyl.} - ethyl) -carbamic benzyl ester According to Example 1 Step 5, by replacing the compound of Step 4 above with (S) - [1 - ((4-benzyloxy-benzyl) - { [(phenyl-cyclobutyl-methyl) -carbamoyl] -methyl} - carbamoyl) -2- (1-trityl-1H-imidazol-4-yl-ethyl] -carbamic acid benzyl ester, the main compound is obtained as a white foam (87% yield) Analysis calculated for C40H42N6O5O.25 DCM, C, 68.28; H, 6.05; N, 11.87, Found: C, 68.23; H, 6.05; N, 11.90.MS - APCI: Calculated for M + 1: 687.3, it was found: 687.2.

EXAMPLE 4 Acid (S) - [1 - ((4-methyl-benzyl) -i [(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl) -carbamoyl) -2- (1H-imidazol-4-yl) ) - ethyl] - carbamic benzyl ester.

Step 1: [(4-methyl-benzyl) -amino] -acetic acid methyl ester. According to Example 1, Step 1, by substituting 4-methyl-benzaldehyde for 4-benzyloxy-benzaldehyde, the main compound is obtained as a colorless oil (53% yield). MS-Cl: Calculated for M + 1: 194; was found: 194 Step 2: Acid (S) -. { (4-methyl-benzyl) - [2-benzyloxy-carbonylamino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -amino) -acetic methyl ester. According to Example 1, Step 2, when replacing the compound of Step 1 above with [(4-benzyloxy-benzyl) -amino] -acetic methyl ester, the main compound is obtained as a white foam (71% yield). MS - APCI: Calculated for M + l: 707.3; of found: 706.3.

Step 3: Acid (S) -. { (4-methyl-benzyl) - [2-benzyloxy-carbonylamino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -annino} - acetic According to Example 1, Step 3, by replacing the compound of Step 2 above with (S) - acid. { (4-benzyloxy-benzyl) - [2-benzyloxy-carbonyl-amino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - Acetic methyl ester, the main compound obtained as a white foam (100% production). MS - APCI: Calculated for M + 1: 693.3; was found: 693.2.

Step 4: Acid (S) - [1 - ((4-methyl-benzyl) -. {[[(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl] -carbamoyl) -2- (1 - trityl-1H-imidazol-4-yl) -ethyl] -carbamic benzyl ester.

According to Example 1, Step 4, by replacing the compound of Step 3 above with (S) - acid. { (4-benzyloxy-benzyl) - [2-benzyloxy-carbonyl-amino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - acetic, the main compound is obtained as a white foam (67% production). MS - APCI: Calculated for M + 1: 836.4; it was found: 836.1.

Step 5: Acid (S) - [1 - ((4-methyl-benzyl) -. {[[(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl] -carbamoyl) -2- (- 1H imidazol-4-yl) -ethyl] -carbamic acid benzyl ester. According to Example 1, Step 5, by replacing the compound of Step 4 with acid (S) [1- ((4-benzyloxy-benzyl) -. {[[(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl] -carbamoyl) -2- (1-trityl-1H-imidazole-4- il) -ethyl] -carbamic benzyl ester, the main compound is obtained as a white foam (55% yield). Analysis calculated for Gs ^ NsO ^ O.D DCM: C, 69.77; H, 6.54; N, 11.58. Found: C, 69.77; H, 6.38; N, 11.52. MS - APCI: Calculated for M + l: 594.3; It was found: 594.2.

EXAMPLE 5 (S) - [1 - ((Benzyloxy-benzyl) -. {[[(1-phenyl-cyclopentyl-methyl) -carbamoyl] -methyl] -carbamoyl) -2 - (- 1H-imidazole - 4-yl) -ethyl] -carbamic acid benzyl ester. According to Example 1, Steps 4 and 5, by substituting C - (1-phenyl-cyclopentyl) -methylamine hydrochloride (U.S. Patent No. 4,755,594; 1988) for C - (1-phenyl-cyclobutyl) hydrochloride - methylamine, 0.28 g (62% over 2 steps) of the main compound is obtained as a white foam. Analysis calculated for C42H44N5? 5'0.5 H2O C, 71.17; H, 6.54; N, 9.88. Found: C, 71.44; H, 6.43; N, 9.93. MS - APCI: Calculated for M + 1: 700.3; was found: 700.3.

EXAMPLE 6 Acid (S) - [1 - ((4-methoxy-benzyl) -. {[[(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl] -carbamoyl) -2- (- 1H- imidazol-4-yl) -ethyl] -carbamic acid benzyl ester. It is synthesized by a variation of Scheme 1 where the trityl protecting group is removed before the formation of the C-terminal amide.

Step 1: [(4-methoxy-benzyl) -amino] acetic acid tert-butyl ester. According to Example 1, Step 1, by substituting 4-methoxy-benzaldehyde for 4-benzyloxy-benzaldehyde and glycine hydrochloride tert-butyl ester for glycine methyl ester hydrochloride, the main compound obtains as a yellow oil (80% strength). production).

Step 2. Acid (S) -. { (4-methoxy-benzyl) - [2-benzyloxy-carbonylamino-3 - (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - acetic tert-butyl ester. According to Example 1, Step 2, upon replacing the compound of Step 1 above, with [(4-benzyloxy-benzyl) -amino] -acetic acid methyl ester, the main compound is obtained as a white foam (73% yield) ).

Step 3: Acid (S) -. { (4-methoxy-benzyl) - [2-benzyloxy-carbonylamino-3 - (1H-imidazol-4-yl) -propionyl] -amino} - acetic To a trityl solution from Step 2 above (3.93 g, 5.14 mmol) in DCM (25 mL) is added TFA (25 mL). The solution is stirred at room temperature for 3 hours and then concentrated. The residue is added to the ether (300 mL) and cooled. The white tarry solid is collected by filtration to give 2.37 g (99% yield) of the main compound.

Step 4: (S) - [1 - ((4-methoxy-benzyl) - { [(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl) -carbamoyl) -2 - (- 1H-imidazole) 4-yl) -ethyl] -carbamic acid benzyl ester. According to Example 1, Step 4, by replacing the compound of Step 3 above, with (S) - acid. { (4-benzyloxy-benzyl) - [2-benzyloxy-carbonyl-amino-3- (1-trityl-1H-imidazol-4-yl) -propionyl] -amino} - acetic, the main compound is obtained as a white foam (17% production). The final purification is achieved by reversible C18 reversible phase by high pressure liquid chromatography using a gradient of 0.1% TFA / H2O.

Analysis calculated for GJ ^ NSOJ- 1.13 TFA -0.4 H2O C, 59.998; H, 5.53; N, 9.39. Found: C, 59.99; H, 5.53; N, 9.26. MS - APCI: Calculated for M + l: 610.3; it was found: 610.3.

EXAMPLE 7 (S) -2- (3-Benzyl-3-methyl-ureido) - N - (4-benzyloxy-benzyl) -3- (1H-imidazol-4-yl) -N- acid. { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} -propionamide. The main compound can be prepared according to Example 1, Step 1, substituting 4-methyl-benzaldehyde for 4-benzyloxy-benzaldehyde; Step 2, replacing N-methyl-N-benzyl-urea-histidine (trityl) (Steps 1 and 2, below) by Cbz - His - (Trt); Step 3, using IN of NaOH in methanol / THF by LiOH: H2O in THF; He passed 4, using DCC / HOBt as binding reagents for BOP; and Step 5, using aqueous acetic acid by 50% TFA in DCM. The main compound is obtained as a white foam; 0.16 g (57% of production).

Analysis calculated for C36H 2NeO3: 0.16 CH2C12: C, 70.01; H, 6.88; N, 13.55. Found: C, 70.04; H, 6.86; N, 13.62. MS - APCI: Calculated for M + 1: 607.8; was found: 607.2.

Step 1: N-methyl-N-benzyl-urea-histidine - (trityl) methyl ester. Histidine hydrochloride - (trityl) methyl ester (2.0 g, 4.2 mmol) is suspended in DCM (20 mL) and the solution is washed twice with saturated NaHCO3, and saline, dried over MgSO4 and cooled to 0 ° C. Triethylamine (0.65 mL, 8.8 mmol) and 4-nitrophenyl chloroformate ( 0.93 g, 4.7 mmol). The reaction is stirred at 0 ° C under nitrogen for 1.5 hours. Then, N-benzyl-N-methylamine (1.14 mL, 8. 8 mmoles) in DCM (10 mL) and the reaction is stirred at room temperature overnight, and ethyl acetate is added to the residue. The organic solution is washed twice with water, saturated NaHC 3, saline and dried over MgSO 4 and concentrated. Chromatography using 1: 1 ethyl acetate: hexanes, produces a foam; 1.19 g (50% of production).

Step 2: N-methyl-N-benzyl-urea-histidine - (trityl) The methyl ester from Step 1 (1.19 g, 2.1 mmol) is dissolved in THF: methanol (10 mL each). NaOH (IN) (6.3 mL, 6.3 mmol) is added and the reaction is stirred overnight. The solvent is removed. HCl (IN) (6.3 mL) is added and the product is extracted with ethyl acetate. The organic solution is then washed twice with saline, dried over MgSO 4 and concentrated to give a white foam; 1.4 g (100% production).

EXAMPLE 8 (S) -2- (3-Benzyl-3-methyl-ureido) - N - (4-benzyloxy-benzyl) -3- (1H-imidazol-4-yl) -N- acid. { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} -propionamide. The main compound can be prepared according to Example 1, Step 2, by substituting N-methyl-N-benzyl-urea-histidine (trityl) (Steps 1 and 2, Example 7), by Cbz-His - (Trt); Step 3, using IN of NaOH in methanol / THF, by LiOH: H2O in THF; Step 4, using DCC / HOBt as binding reagents for BOP; and Step 5, using 80% aqueous acetic acid by 50% TFA in DCM. The main compound is obtained as a white foam; 0.12 g (52% of production).

Analysis calculated for C42H 6N6O4: 0.33 CH2C12: C, 68.87; H, 6.33; N, 11.39. Found: C, 68.87; H, 6.43; N, 11.11.

MS - APCI: Calculated for M + l: 699.9; it was found: 699.4.

EXAMPLE 9 Acid (S) - [1 - [(4-benzyloxy-benzyl) - ( { [1 - (2,6-dichloro-phenyl) -cyclobutyl-methyl-carbamoyl.) - methyl) -carbamoyl ] - 2 - (1H-imidazol-4-yl) -ethyl] -carbamic methyl ester. The main compound can be prepared according to Example 1, Step 3, using IN of NaOH in methanol / THF by LiOH: H2? in THF; Step 4, substituting [1- (2,6-dichlorophenyl) cyclobutyl] methylamine hydrochloride (Step 1 below) for C - (1-phenyl-cyclobutyl) methylamine hydrochloride. The main compound is purified by reversible phase of high pressure liquid chromatography (eluent: 0.1% aqueous TFA and 0.1% TFA in acetonitrile) to give a white foam; 0.015 g (5% of production).

Analysis calculated for 1.68 C2HtO2F3: 1.01 H2O: C, 55.36; H, 4.47; N, 7.28. It was found: C, 55.36; H, 4.47; N, 7.26. MS - APCI: Calculated for M + l: 752.2; it was found: 752.6.

Step 1: [1- (2,6-Dichlorophenyl) cyclobutyl] methylamine hydrochloride. The compound 1- (2,6-dichloro-phenyl) -1-cyclobutanecarbonitrile (1 g, 4.4 mmol) is reduced with Raney nickel in methanol / NH3. The catalyst is removed and washed with methanol. The filtrate is concentrated and diethyl ether (100 mL) is added to the residue. Concentrated HCl is added dropwise to precipitate the desired product; 1.05 g (100% production).

EXAMPLE 10 [(S) -Trans] - [1 - (But-2-enyl - ([(1-phenyl-cyclobutyl-methyl) -carbamoyl] -methyl] -carbamoyl) -2- (3H-imidazole) acid - 4 - yl) - ethyl] - carbamic benzyl ester Step 1: Methyl 2 - [(E) -2-butenylamino] acetate A suspension of (E) -2-buten-1-amine HCl (5.37 g, 49.9 mmoles) (Chem. Ver., 1984; 117: 1250) in acetonitrile (100 mL) is treated with methyl bromoacetate (4.72 mL, 49.9 mmol) and Et3N (14.0 mL, 99.8 mmol) and stirred at room temperature. After the suspension is heated to reflux overnight, the solution occurs at the reflux temperature.After cooling, the Et3N? Cl precipitate is filtered and the solvent is removed under reduced pressure leaving 5.0 g of the product. crude chromatography on silica gel, eluting with CHGs / MeOH (98: 2) gives 1.41 g (19.8% yield) of the pure product as an oil.

Step 2: Methyl 2-Í2 - [(benzyloxy) carbonyl] amino-3 - (1-trityl-1H-5-imidazolyl) propanoyl] [(E) -2-butenyl] aminoacetate. A solution of methyl 2 - [(E) -2-butenylamino] acetate (from step 1) (0.6 g, 4.2 mmol) in CH2C12 (50 mL) is cooled in ice and treated with Z-His (Trt) (2.23). g, 4.2 mmol), diisopropyl-ethyl-amine (2.2 mL, 12.6 mmol), and PyBOP (2.2 g, 4.2 mmol). After stirring at 0 ° C for 15 minutes, the solution is stirred at room temperature for 4 days. After removing the solvent under reduced pressure, the residue is fixed in EtOAc, washed three times with H2O and then with saturated NaCl. It is dried over MgSO 4 and the solvent is removed under reduced pressure and 4.36 g of crude product remain. Chromatography on silica gel, eluting with CHCIj / MeOH (98: 2) gives 2.23 g (81.1% yield) of the pure product as a white solid foam.

MS, m / z 657 (M + H.

Step 3: 2 - [2 - [(Benzyloxy) carbonyl] amino-3- (1-trityl-1H-5-imidazolyl) propanoyl] [(E) -2-butenyl] aminoacetic acid. To an ester solution (from Step 2) (2.23 g, 3.4 mmol) in MeOH / dioxane (20 mL and 15 mL), respectively) are treated with 2N NaOH (7.0 mL, 14.0 mmol) and stirred at room temperature. the room for 0.5 hours. After adding 2N HCl (7.0 mL, 14.0 mmol), the mixture remains in a solid. This solid is mixed with EtOAc / THF and filtered to remove the NaCl. Removal of the solvent under reduced pressure leaves 2.06 g (94.5% yield) of the product as a white solid foam. MS, m / z 643 (M + H +).

Step 4: Benzyl N-2 - [(E) -2-butenyl (2-oxo-2 - [(1-phenyl-cyclobutyl) methyl] aminoethyl) amino] -2-oxo -! - [(! - trityl-1H-4-imidazolyl) methyl] ethylcarbamate. The compound (1 g, 1.6 mmol) from Step 3 is dissolved in methylene chloride (50 mL), HOBt (0.29 g, 2.1 mmol) followed by 0.5 M DCC / DCM (3.8 mL, 1.9 mmol) is added, C - (1-phenyl-cyclobutyl) -methylamine hydrochloride (0.37 g, 1.9 mmol) and DIEA (0.61 mL, 3.5 mmol). The reaction is stirred at room temperature, under a nitrogen atmosphere for 2 days. The solution is filtered and the filtrate is concentrated under reduced pressure. The residue is fixed in ethyl acetate and the organic solution is washed twice with 2N HCl, INN NaHC 3 and saline, dried over MgSO 4, filtered and concentrated. Chromatography on silica gel, eluting with CHCl3 / CH3OH (95: 5) gives 0.97 g (77% yield) of the pure product.

Step 5: [(S) -trans] - [1 - (but-2-enyl - ([(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (3H-imidazole- 4-yl) -ethyl] -carbamic acid benzyl ester.

The main compound is obtained by treating the compound from Step 4 (0.97 g, 1.2 mmol) with glacial acetic acid (20 mL) and water (5 mL) at reflux for 40 minutes. The reaction mixture is cooled and concentrated under reduced pressure. The residue is fixed in ethyl acetate and the organic solution is washed four times with saturated NaHC 3, twice with saline, dried over MgSO 4, filtered and concentrated. Chromatography with silica gel, eluting with CHCl3 / CH3OH (0.6% CH3OH in CHCl3) gives 0.43 g (66% yield) of the pure product.

Analysis calculated for C3tH37N5? 4: 0.25 CHC13: C, 65.45; H, 6.55; N, 12.21. It was found: C, 65.59; H, 6.61; N, 12.20. MS - APCI: Calculated for M + 1: 544.7; it was found: 544.3.

EXAMPLE 11 Acid (S) - [2 - (3H-imidazol-4-yl) -1- ( { [(1-phenyl-cyclobutyl-methyl) -carbamoyl-methyl.} - propyl-carbamoyl) -aryl ] - carbamic benzyl ester. The main compound can be prepared according to Example 10, by performing Step 1 as shown below. The main compound is obtained as a white foam; 0.25 g, (66% production).

Analysis calculated for C3oH37N5? 4: 0.10 CHC13: C, 66.51; H, 6.88; N, 12.88. Found: C, 66.52; H, 6.84; N, 13.11. MS - APCI: Calculated for M + l: 532.7; it was found: 532.2.

Step 1: Methyl 2 - (propylamino) acetate A suspension of glycine methyl ester • HCl (5.0 g, 39.8 mmol) in acetonitrile (100 mL) is treated with propyl bromide (3.7 mL, 39.8 mmol) and diisopropyl ethyl amine (13.9 mL, 79.6 mmol). After stirring at room temperature for 1 hour, the mixture is heated to reflux overnight. The solution occurs at reflux temperature. The solvent is removed under reduced pressure and the residue is triturated with Et2? / EtOAc. After filtering and removing the solvent under reduced pressure, 1.13 g of the crude product remain.

Chromatography on silica gel, eluting with CHCIj / MeOH (98: 2) gives 0.53 g (10.2% yield of the product as a yellow oil, MS, m / z (M + H +).

EXAMPLE 12 Benzyl N - ((SS) - 1 - (1 H - 4 - imidazolylmethyl) -2-oxo-2 - (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) [(IR) - 1-phenyl-ethyl] aminoethyl] carbamate The main compound is prepared according to Example 10, Step 1, substituting R-a-methyl-benzyl-amine for (E) -2-buten-1-amine • HCl; 2, using HATU by PyBOP, step 3, using LiOH: H2? In THF by 2N NaOH in methanol / dioxane The main compound is obtained as a white foam, 0.51 g (66% yield).

Analysis calculated for C35H39N5O4: 0.35 CH2C12: C, 68.10; H, 6.42; N, 11.23. It was found: C, 68.25; H, 6.46; N, 11.23. MS - APCI: Calculated for M + l: 594.7; was found: 594.2.

EXAMPLE 13 Acid (S) - [1 - ((1-dimethyl-2-phenyl-ethyl) - ([(1-phenyl-cyclobutyl-methyl) - carbamoyl] - methyl} - carbamoyl) -2- (3H-imidazol-4-yl) -ethyl] -carbamic acid benzyl ester. The main compound can be prepared according to Example 10, Step 1, substituting β, β-dimethyl-phenethyl-amine hydrochloride (Step A, below) for (E) -2-buten-1-amine • HCl; Step 2, using HATU and HOAT by PyBOP; Step 4, using PyBOP by DCC / HOBt; Step 5, using 95% TFA in DCM for glacial acetic acid in water. The main compound is obtained as a white foam; 0.121 g (12% of production).

Analysis calculated for H42N5? 4: 0.33 CH2C12: C, 69.11; H, 6.63; N, 10.79. It was found: C, 69.10; H, 6.91; N, 11.00 MS - APCI: Calculated for M + l: 621.8; it was found: 622.2.

Step A: ß. ß-dimethyl-phenethyl-amine by hydrochloride A solution of sodium hydride (60% in 17 g oil, 0.43 mmol) is suspended in THF (150 mL) and cooled to 0 ° C under nitrogen. Benzyl cyanide (22.2 g, 0.19 mmol) in THF (30 mL) is added dropwise and the reaction is stirred for 1 hour. Iodomethane (24.9 mL, 0.4 mmol) in THF (20 mL) is added dropwise at 0 ° C. The reaction is stirred at room temperature overnight, under nitrogen. The solution is filtered and the filtrate removed in vacuo. The residue is fixed in ethyl acetate (100 mL) and washed three times with 10% NaHS? 3, saturated NaHCO3, saline and dried over MgSO4, concentrated; 22.74 g (92% of production).

The reduction of the above product is carried out in the presence of Raney nickel, in methanol / NHa. The catalyst is removed and washed with methanol. The filtrate is concentrated and diethyl ether (100 mL) is added to the residue. Concentrated HCl is added dropwise to precipitate the desired product; 24.8 g, (86% of production).

Inhibitory activity of PFT. The inhibitory activity of the protein: farnesyltransferase (PFT) or farnesyl protein transferase (FPT) of the compounds of the present invention were tested in HEPES regulator (pH 7.4) containing 5 mM potassium phosphate and 20 μM ZnCl2. The solution is also contained in 5 mM DTT (dithiothreitol), 5 mM MgCl2 and 0.1% PEG 8000. The experiments were carried out in 96-well plates (Wallec) and solutions composed of varying concentrations of a compound of the present invention in 100% DMSO (dimethylsulfoxide) were employed. During the addition of both substrates, the specific activity 15-30 Ci / mmol of farnesyl pyrophosphate ([1 - 3H], final concentration of 134 nM) and (biotinyl) - Ahe - Thr - Lys - Cys - Val - is radioidentified. He-Met ([3aS [3a alpha, 4 beta, 6a alpha] - hexahydro-2-oxo-1H-thieno [3,4-d] 5 -pentanoic acid] - [7-amino-heptanoic acid] - Thr - Lys - Cys - Val - He - Met) (Ahe is 7 - amino - heptanoic acid, Thr is threonine, Lys is lysine, Cys is cysteine, Val is valine, He is isoleucine and Met is methionine) (final concentration 0.2 μM) , the reaction of the enzyme is initiated by the addition of purified rat farnesyl protein transferase with affinity to SF9. After incubation at 30 ° C for 30 minutes, the reaction is terminated by diluting the reaction to 2.5 times with a stop buffer containing 1.5 M of magnesium acetate, 0.2 M of HJPO4, BSA (serum albumin of bovine) at 0.5% and strepavidin beads (Amersham) at a concentration of 1.3 mg / mL. After allowing the plate to settle for 30 minutes, at room temperature, the radioactivity is quantified on a microBeta counter (Model 1450, Wallec). The experiment is carried out without the 5 mM potassium phosphate.

Gel change experiment. Twenty-four hours after planting 2 x 106 ras-transformed cells per treatment condition, the farnesylation inhibitor is added at varying concentrations. After 18 hours of the incubation period, cells are lysed in a phosphate buffered saline solution containing 1% Triton X-100, 0.5% sodium deoxycholate and 0.1% SDS (sodium dodecyl sulfate), pH 7.4 in the presence of several protease inhibitors (PMSF (phenyl-methyl-sulfonyl fluoride), analgesics, leupeptin, pepstatin A and aprotinin, all at 1 μg / mL). The ras protein is immunoprecipitated from the supernatants by the addition of 3 μg v-H-ras Ab-2 (Yl 3-259 antibody from Oncogene Science). After overnight immunoprecipitation, 30 μL of a 50% G-sepharose protein mixture (Pharmacia) is added, followed by incubation for 45 minutes. The pellets are resuspended in a 2X tris-glycine charge regulator (from Novex) containing 5% β-mercapto ethanol and then denatured for 5 minutes by boiling before electrophoresis in 14% SDS Tris-glycine gel. Using transfer techniques to the west, the proteins are transferred to nitrocellulose membranes followed by blocking in the blocking regulator. During the incubation at night with primary antibodies (Pan - ras Ab - 2 of Oncogene Science), a HRP is used. (horseradish peroxidase) of anti-mouse conjugated secondary antibody (Amersham), for the detection of ras protein. Stains develop using ECL techniques (increased chemiluminescence) (Amersham).

Clonogenic Experiment (6-well plates) Activities previously done to establish a current test 1. Make Bacto 1.5% agar in Milli-Q water and autoclave. 2. Make 500 mL of 2X DMEM - HG without red phenol by combining the following: 1 bottle of DMEM base powder. (Sigma D - 5030) 4.5 g of glucose 3.7 g of sodium bicarbonate 0.11 g of sodium pyruvate 20 mL of 200 mM of L - glutamine (Sigma G - 7513) 1 mL of pen - strep (GibcoBRL No. 15140-023 ) Adjust the pH to 7.1 with HCl; filter, sterilize. 1. Assemble the water bath that makes the change (water precipitation basin with thermometer, on hot plate) in the lid. Maintain the water temperature between 37 ° and 43 ° C. 2. Bacto Agar 1.5% autoclave for approximately 2 minutes at high level, or until melting is complete. Then let cool a little before using (you can prevent it from solidifying again by placing the bottle on the hot plate).

Bottom agar layer (0.6%) Top layer (3% agar) 20% calf serum 20% calf serum 2X DMEM 40% 2X DMEM 50% Bacto agar 40% (1.5%) Bacto 20% agar (1.5%) Sterile water x 10 μL at 10% Cell suspension (for = 5000 cells / well) (H61 cells: NIH transformed 3T3 NIH cells) Depending on the volume that each layer needs, use 50 mL conical tubes or 200 mL turnip tubes, which can float in the "water bath". 4. Add 1 mL of agar medium from the bottom layer to each well; put 1 mL of warm agar medium in a well; then, using the tip of a pipette, spread the agar medium around to completely cover the bottom. Repeat with the next well. Do not add the last mL in the pipette to a well, make bubbles.

. Place the plates at room temperature for 5 minutes until the bottom layer solidifies. 6. Identify Falcon 2054 tubes (12 x 75 mm) and add the appropriate volume of drug solutions within them. 7. Aliquot 4 μL of DMSO or the solution with the drug per 1 mL of agar medium to the appropriate tubes, then add agar / medium / cells to each tube. Always add 1 mL more than what you really need. Mix and lower the pipette (gently); then, place 1 mL at the center of each well. The upper layer is less viscous, so it will generally spread on the bottom without help. If necessary, rotate the plane of the plate gently to spread the top layer evenly over the bottom layer. 8. Leave the plate for 5 to 10 minutes at room temperature to solidify, then put it in a 37 ° C incubator with 5% CO2. 9. On day 13, add 0.5 mL of INT (tetrazolium 1 mg / mL in filtered and sterilized Milli-Q water) and return the plates to the incubator.

. Count the colonies.

The information in the following table shows the activity of the farnesyl protein transferase inhibitory activity, activity in the gel change experiment, and the activity in the Clonogenic experiment of the compounds of the present invention.

M. E. D.: is the minimum effective dose to observe inhibition of ras farnesylation # NT = Not Tested.

In vivo activity The NCR-A-female mice were taken at random and inoculated with trocar fragments of xenografts obtained from donor animals on Day 0 of the experiment. The H61 cells are NIH3T3 cells that have been transfected and transformed with human mutant H-ras, then the animals were randomized in the treatment groups. Mice that had tumors were treated via subcutaneous (SC) or intraperitoneal (IP) injection of Example 1 every 12 hours at different dose levels for 14 days starting on day 1. Tumors were measured with two-dimensional orthogonal gauge several times per week for the duration of the experiment. The tumor loads in milligrams are estimated from calibrator measurements by standard methods (mg = axb2 / 2, where a and b are the measurements of the major and minor axes, respectively). The inhibition of tumor growth is established on the last day of treatment (14). Example 1 significantly inhibits tumor growth in this experiment. When administered subcutaneously at 96 mg / kg / injection, the median tumor burden of the treated group is 672 mg, compared to a median load of 2508 mg per vehicle treated in the control group, representing a 73% inhibition of tumor growth. .

In mice that are administered intraperitoneally in Example 1, either at 37 or 23 mg / kg / injection, there are no obvious tumors at the inoculation sites, indicating complete (100%) inhibition of tumor growth.

Claims (2)

CLAIMS A compound that has Fúuula I. where: RQ is ~ Cg alkyl? XesOo 1; each R '"* is independently hydrogen or G - ß aiquiio; A is - COR", -: > CO2, - Cs > , O S II II - CSR ", - CNR'R" ", - C (S) OR" \ - C (S) NHR "\ - SO2R", or - CONR "R \ IC, R "and l <" '"are independently Cj-ß alkyl, - (CR? U) m-cicioalquüo, 0 -. 0 - (CR'Vjn, - aryl, or - (CR'V) ,,, - heteroaryl; Each in is indepenieiilemeule 0 to 3, R !, R \ and R4 are independently hydrogen or C, -C alkyl. (\ RJ cs - í Cí Í > ", Cí? (Clh. G -C? Aiauiio. I2 -, aiqueuu, (- naphthyl, - (H2), Ü2R ', - (< -H2 NR R, - (CH2). - O -Ci - CG alkyl, - (CH2) t - OH - (-i2 - monomial \ • fij \ M 14 1 N C (Cr! 2) v ..14 \ / V y - (CR'4JLiR! 4) m - (ténil substituted with Rr), or - (CR! 4R! 4) m- (heteroap i substituted with Rb); t is 2 to 6, v is I to 6 R "is - - tinnil, - O - benzyl, halogen, G - alkyl, hydrogen, - O G - C alkyl, - NH2, - NHR \ - NRaR \ O O O II II II 11 ^ II II - CCi - Ce alkyl, - C - aryl, - OH, - CFj, - NO2, - COH, O O II II II II - COC, - Ce alkyl, - CN, - OPOjH2, - CH2PO3H2, - CO mil, II II II II - -It t2, I H, - NR R, - R -K, -U (H? JyiNR, - Neither. - CF2CF3, - SQ2R \ - SO2NRaRa \ - CHO, -OCOCII ?. - u < r? 2jm - neiei? alti, -? iLrijjm - aip, - (Cr? 2jm - uu? aiquii, / r * tt \ 1, -i rrtv p? rr - (? _ ??, / m - aeieioaiu, or -,? - ?? or .5, and it's 2 or 5, R5 is R "and R) (Tenyl substituted with Rft, R "and R each // is iiidependienlepte 2, 3 or 4, R ', RS and Rh are independently hydrogen, halogen, -? or II II - L.I 2 u3? i2. - u - ienil, - O -? enzn, - L IN? 2, - LINIÍR O O II ll tlNK K, - Í 11 -, - lCn) yl K K, - 1NÍ1, lNíIR, U O II II. NR-R-,. cCi - Ce - alkyl, - C - aryl. - OH, - CF3. - NO2, - 'J KJ II II II - ton, - * -. Ov.-? - e aiquilo, - Cu aru, - IN3, - CF2 F3, - SU2. -? L > 2l, - K..HS, O - ÜL-U -íl ?; > ' R and Rd are independently Ci-C6 alkyl, -lhjm-cycloalkyl or liiuiógeu ?, and the salts, sleí is, acceptable amides and pi? di? gas faimacéulicauíeiile of l? s same. 2. a compound according to Claim i where: R1 is hydrogen, R2 is hydrogen, R4 is hydrogen, R14 is hydrogen or etui, and Á In? Li? ejemμlai foot ilect? of i? s c? mpuesl? s of F? nula I. C2 - ß alkenyl, - (CH2) m - (phenyi substituted with R) - (U2) m - (heieroapl substituted with R "). R 'is hydrogen, R "is hydrogen, R' is hydrogen and R" is hydrogen or methyl H. a compound that acueruo a? A? E? V? Nu? Cac? N i u? Nue. is (CH) n -CH., C ((substituted lfeeiniuil c? N fiu? I ?, cl? I ?, bi? M? O NH2 at position-2), or (CH2) n '"H- O -pipun. They are also included in the Compounds that have the Form II. ?? ID where rr, - rr i -i L (, aiqun? R6 cs - O - bcnzil, - OCH - phenyl, - NH - benzif 0 - I ^ - I -., (Aiquiio) - benzii, or -? H2 - lenu, C is hydrogen, halogen, G - C aiquüo, - - benzil, -? r > Knrti? - p -ip-u1n-1, - r rr ra _ ?? quu -i? - t rrrrj, - «JXHJ, l Ce-im '' J .I - -e, R1" and R "are independently hydrogen or G-Ce alkyl, each n is independently 2 or 4 R "is R" is hydrogen or meth; R1, R ': and R' are independently hydrogen, halogen, - NH2, - NHR ", - OG - Ce alkyl ?, or - G - Ce alkyl and the salts, esteri is, amidas and pi? di? gas faimacéulicamenle acepiames ue ios ms. 6. A c? Mpuest? that has the F? iula III. where: caaa ¡i is _, J o -t; X is N ?? O, or - NCH3; R 1 is -O-benzyl, -Cl hydrogen, halogen, -OH, -phenyl, -G-C alkyl, -0CH 2 -pyridyl, or -OG-Cr, alkyl and pharmaceutically acceptable salts, esters, amides and prodrugs of L? s more. 7. a compound that has F? iula IV. where: X is NH, O, or -NCH3; R14 is hydrogen or G-Cβ alkyl; Rffl is hydrogen or G-C6 alkyl, C2-Ce alkenyl, - ((CH2 NR14R14, - (CH2) V-O-G-Ce-alkyl, - (CH2-OH, - (CH2) t-morpholino, - ( CH2t NN- R14, (CH2) VN R14 '- (CH2) v - CO2R14, or - (CH2) m CO (CH2; n is 2, 3 or 4; m is 0 to 3; t is 2 to 6; It's 1 to 6. 8. A pharmaceutically acceptable composition comprising a compound of Claim 1. 9. A pharmaceutically acceptable composition comprising a compound of Claim 5. 10. A pharmaceutically acceptable composition comprising a compound of Claim 6. 11. A method for treating or preventing restenosis, the method comprises administering to a patient having restenosis or a risk of restenosis a therapeutically effective amount of a compound of Claim 1. 12. A method for treating or preventing restenosis, the method comprises administering to a patient having restenosis or a risk of restenosis a therapeutically effective amount of a compound of Claim 5. 13. A method for treating or preventing restenosis, the method comprises administering to a patient having restenosis or a risk of restenosis a therapeutically effective amount of a compound of Claim 6. 14. A method for treating cancer, the method comprises administering to a patient having cancer, a therapeutically effective amount of a compound of the Claim 1 15. A method for treating cancer, the method comprises administering to a patient having cancer, a therapeutically effective amount of a compound of Claim 5. 16. A method for treating cancer, the method comprises administering to a patient having cancer, a therapeutically effective amount of a compound of Claim 6. 17. The compounds: (S) - [1 - ((4-benzyloxy-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazole - 4-yl) -ethyl] -carbamic acid benzyl ester; (S) - [1 - ((4-Benzyloxy-benzyl) -. {[[(1-phenyl-cyclopropylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - [1 - ((4-Benzyloxy-benzyl) -. {[[(1-phenyl-cyclopentylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - [1 - ((4-Phenyl-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} - carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - [1 - ((4-methoxy-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - [1 - ((4-Methyl-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} - carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - carbamic benzyl ester; (S) - N - (4-Benzyloxy-benzyl) -2- (3-benzyl-ureido) -3- (1 H -imidazol-4-yl) -N- acid. { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} -propionamide; (S) -2- (3-Benzyl-3-methyl-ureido) -N - (4-benzyloxy-benzyl) -3- (1H-imidazol-4-yl) -N- acid. { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} -propionamide; . (S) - [1 - ((4-Benzyloxy-benzyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) acid ) - ethyl] - thiocarbamic S -benzyl ester; (S) - (2 - (1H-imidazol-4-yl) -1- {. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] - [4 - (pyridine-2) acid - il-methoxy) -benzyl] -carbamoyl.} - ethyl) -carbamic benzyl ester; (S) - [1 - ((Cyclohexyl-methyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2 - (1H-imidazol-4-yl) - acid ethyl] - carbamic benzyl ester; (S) - (1 - ((isobutyl) - { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl.} - carbamoyl) -2 - (1H-imidazol-4-yl) -ethyl] acid] - Carbamic S - benzyl ester. 18. The compounds: (S) -2- (3-Benzyl-3-methyl-ureido) -3- (1H-imidazol-4-yl) -N (4-methyl-benzyl) -N- acid. { [(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} propionamide; (S) - [1 - [(4-Benzyloxy-benzyl) - ( { [1 - (2,6-dichloro-phenyl) -cyclobutylmethyl] -carbamoyl} -methyl) -carbamoyl] -2- (1H) acid imidazol-4-yl) -ethyl] carbamic acid benzyl ester; [(S) -trans] - [1 - (But-2-enyl - {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} - carbamoyl) -2 - (3H-imidazole-4) acid - il) - ethyl] - carbamic benzyl ester; (S) - (2 - (3H-imidazol-4-yl) -1 - ( { [1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl} - propylcarbamoyl) -ethyl] -carbamic acid benzyl ester ester; Benzyl N - ((1S) -1- (1H-4-imidazolylmethyl) -2-oxo-2 - (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) [(IR) -1-phenylethyl ] aminoethyl) carbamate; (S) - [1 - ((1,1-dimethyl-2-phenyl-ethyl) -. {[[(1-phenyl-cyclobutylmethyl) -carbamoyl] -methyl] -carbamoyl) -2- (3H) acid imidazol-4-yl) -ethyl] -carbamic acid benzyl ester. 19. The compounds: Benzyl N - [(SS) -2 - [(2-hydroxyethyl) (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) amino] -1 - (1H-4-imidazolylmethyl) -2 oxoethyl] carbamate; 3 - [[(2S) -2 - [(benzyloxy) carbonyl] amino-3 - (lH-4-imidazolyl) propanoyl] (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] propanic; Methyl-3 - [[(2S) -2 - [(benzyloxy) carbonyl] amino-3 - (1H-4-imidazolyl) propanoyl] (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino propanoate; Benzyl N - [(SS) -2 - [(2-aminoxyethyl) (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) amino] -1- (1H-4-imidazolylmethyl) -2-oxoethyl] carbamate; Benzyl N - (IS) - (1H-4-imidazolylmethyl) -2 - [[2- (methylamino) ethyl] (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] -2-oxoethylcarbamate; Benzyl N - (IS) - (1H-4-imidazolylmethyl) -2 - [(2 - (methoxyethyl) (2-oxo-2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] -2-oxoethylcarbamate; Benzyl N - (IS) - (1H-4-imidazolylmethyl) -2 - [(2 - (morpholinoethyl) (2-oxo-2) - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] -2-oxoethylcarbamate; Benzyl N - (lS) - (lH-4-imidazolylmethyl) -2 - [(2 - (methyl-2-phenylpropyl) (2-oxo-2 - [(1-phenylcyclobutyl) methyl] aminoethyl) amino] -2- oxoethylcarbamate; and

1 - . 1-Phenylethyl N - [(SS) -

2 - [[4- (benzyloxy) -1,5-cyclohexadienyl] methyl (2-oxo) - 2 - [(1-phenylcyclobutyl) -methyl] aminoethyl) amino] -1 - (1H-4-imidazolylmethyl) -2-oxoethyl] carbamate. 20. The method of Claim 13 wherein the cancer is lung cancer, colon cancer, breast cancer, pancreatic cancer, thyroid cancer or bladder cancer. 21. A method for treating psoriasis, the method comprises administering to a patient having psoriasis a therapeutically effective amount of a compound of Claim 1. 22. A method for treating psoriasis, the method comprises administering to a patient having psoriasis a therapeutically effective amount of a compound of Claim 5. 23. A method for treating psoriasis, the method comprises administering to a patient having psoriasis a therapeutically effective amount of a compound of Claim 6.