CA2249615A1 - Inhibitors of farnesyl-protein transferase - Google Patents
Inhibitors of farnesyl-protein transferase Download PDFInfo
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- CA2249615A1 CA2249615A1 CA002249615A CA2249615A CA2249615A1 CA 2249615 A1 CA2249615 A1 CA 2249615A1 CA 002249615 A CA002249615 A CA 002249615A CA 2249615 A CA2249615 A CA 2249615A CA 2249615 A1 CA2249615 A1 CA 2249615A1
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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
CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 TITLE OF THE INVENTION
~NHIBITORS OF FARNESYL-PROTEIN TRANSFERASE
S BACKGROUND OF THE ~VENTTON
The present invention relates to compounds which inhibit farnesyl protein transferase, a protein which is implicated in the oncogenic pathway mediated by Ras. The Ras proteins (Ha-Ras, Ki4a-Ras, Ki4b-Ra.s and N-Ras) are part of a signalling pathway that links cell surface growth factor receptors to nuclear signal.s initiating cellular proliferation. Biological and biochemical studies of Ras action indicate that Ras functions like a G-regulatory protein. In the inactive state, Ras is bound to GDP. Upon growth factor receptor activation Ras is induced to exchange GDP for GTP and undergoes a conformational change. The GTP-bound form of Ras propagates the growth stimulatory signal until the signal is termin~ted by the intrinsic GTPase activity of Ra,s, which returns the protein to its inactive GDP
bound form (D.R. Lowy and D.M. Willumsen, Ann. Rev. Biochem.
62:~51-~91 (1993)). Mutated ras genes (Ha-ras, Ki4a-)as, 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 ~re involved with Ras membrane localization, and all 3 modifications occur at the C-terminus of ~as. The Ras C-terminu.s contain,s a sequence motif termed a "CAAX" or "Cy.s-Aaal-Aaa2-Xaa"
box (Cys is cy~steine, Aaa is an aliphatic amino acid, the Xaa is any - 30 amino acid) (Willumsen et al., Natu) ~ 310:5~3-5~6 (19~4)). Depend-ing on the specific .sequence, this motif serves as a signal .sequence for - the enzymes farnesyl-protein transfera.se or geranylgeranyl-protein transferase, which catalyze the alkylation of the cysteine residue of the CAAX motif with a Cls or C2() isoprenoid, respectively. (S. Clarke., CA 0224961~ 1998-09-22 WO 97/36876 PCT/US97/~6257 Ann. Pev. Bi~chen~ 355-386 (1992); W.R. Schafer and J. Rine, Ann. Rel~. Genetics 30:209-237 (1992)). Ras proteins are known to undergo post-translational farnesylation. Other farnesylated proteins include the Ras-related GTP-binding proteins such as Rho, fungal 5 mating factors, the nuclear lamins, and the gamma subunit of transducin. James, et al., J. Biol. Chem. 269, 14182 (1994) have identified a peroxisome associated protein Pxf which is also farnesylated. James, et al., have also suggested that there are farnesylated proteins of unknown ~structure and function in addition 10 to those listed above.
lnhibition 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 demonstrated that certain inhibitors of farnesyl-protein transferase 15 selectively block the processing of the Ras oncoprotein intracellularly (N.E. Kohl et al., Science, 260:1934-1937 (1993) and G.L. James e~ al., Science, 260:1937-1942 (1993). Recently, it has been shown that an inhibitor of farnesyl-protein transferase blocks the growth of ~as-dependent tumors in nude mice (N.E. Kohl et al., Proc. Natl. Acad.
20 Sci U.S.A., 91:9141-9145 (1994) and induces regression of mammary and salivary carcinomas in ras transgenic mice (N.E. Kohl et al..
Nature Medicine, 1 :792-797 (1995).
Indirect inhibition of farnesyl-protein transfera~se in l'iVO
has been demonstrated with lovastatin (Merck & Co., Rahway, NJ) 25 and compactin (Hancock et al., ibid; Casey et al., ibid; Schafer et al., Science 245:379 (l9~S9)). The.se drugs inhibit HMG-CoA reductase, the rate limiting enzyme for the production of polyisoprenoids including farnesyl pyrophosphate. Farnesyl-protein transferase utilizes farnesyl pyrophosphate to covalently modify the Cys thiol group of the Ras 30 CAAX box with a farnesyl group (Reiss et al., Cell 62:~ g (1990);
Schaber et al., J. Biol. Chem., 2~5:14701-14704 (1990); Schafer et al., - Science, 249: 1133- 1139 (1990); Manne eF al., Pro( . Natl. Acad. Sci USA, 87:7541-7545 (1990)). Inhibition of farnesyl pyropho~phate biosynthesis by inhibiting HMG-CoA reductase blocks Ras membrane CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 Iocalization in cultured cells. However, direct inhibition of farnesyl-protein tran~sferase would be more specific, and thus preferable.
Inhibitor.s of farnesyl-protein transferase (FPTase) have been described in two general classes. The first are analogs of farnesyl 5 diphosphate (FPP), while the second class of inhibitors i~ related to the protein ~ubstrates (e.g., Ras) for the enzyme. The peptide derived 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., ihid; Reiss et. al., ihid; Reiss 0 et al., PNAS, 88:732-736 (1991)). Such inhibitors may inhibit protein prenylation while serving as alternate substrates for the farnesyl-protein tran,sferase 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)).
It has recently been reported that FPT-ase inhibitor,s also inhibit the proliferation of vascular smooth muscle cells and are therefore useful in the prevention and treatment of arteriosclerosi,s and diabetic disturbance of blood ves,sels (JP H7-112930).
It has recently been di,sclosed that certain tricyclic 20 compounds which optionally incorporate a piperidine moiety are inhibitors of FPTase (WO 95/10514, WO 95/1051~ and WO 95/10516).
Imidazole-containing inhibitors of farne~yl protein transferase have also been disclosed (WO 95/09001 and EP 0 675 112 Al).
The present invention addresses a compound of formula I:
V A1(CRla2) A2(CR'b2)n ~(W)~ (CR 2t~A - (CR 2)p or a pharmaceutically acceptable ~alt thereof, wherein:
CA 0224961~ 1998-09-22 W 097/36876 PCTrUS97/06257 Rla, R1b and R2 are independently selected from the group consisting of: hydrogen? aryl, substituted aryl, heterocyclyl, C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R~O-, R9S(o)m wherein m is 0, 1 or 2, R~C(O)NR8-, CN, NO2, (R8)2NC(NR8)-, R8C(O)-, R80C(O)-, N3, -N(R8)2, R90C(o)NR8- and Cl-C6 alkyl, unsubstituted or substituted by 1-3 groups .selected from the group consisting of: halo, aryl, heterocyclyl, C3-clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(o)m-~ R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R~)2 and 1 0 R9OC(o)NR8-;
R3 and R4 are independently selected from the group consisting of: H, F, Cl, Br, -NR82, CF3, NO2, R8O-, R9S(~)m-~
CF3(CH2)n-O-, R8C(O)NH-, H2NC(NH)-, R8C(O)-, R8OC(O)-, N3, CN, R9OC(o)NR8-, C1-C20 alkyl, substituted or unsubstituted aryl and ,substituted or unsubstituted heterocyclyl;
A3 is selected from: --C----C , --R8C=CR8---C(O)-, aryl, heteroaryl or a bond;
provided that when A3 is heteroaryl, attachment of A3 the remainder of the molecule is through substitutable heteroaryl ring carbon,s;
X represents aryl or heteroaryl;
provided that when X is heteroaryl, attachment of X the remainder of the molecule is through substitutable heteroaryl ring carbons;
R6 is independently selected from the group consisting of:
hydrogen, aryl, heterocyclyl, C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-6 perfluoroalkyl, F, Cl, Br, R~O-, R9S(o)m-~
R8C(O)NR8-, CN, NO2, (R8)2Nc(NR8)-~ R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(o)NR8- and Cl-C6 alkyl unsubstituted or substituted by 1-3 group~s selected from: aryl, heterocyclyl, C3-Clo cycloalkyl, CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/06257 C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R~O-, R9S(o)m-~ R8C(O)NR~-, CN, (R~)2NC(NR~)-, R~C(O)-, R~OC(O)-, N3, -N(R~)2 and R9OC(o)NR~-;
R7 is independently selected from the group consisting of:
hydrogen, aryl, heterocyclyl, C3-C1o cycloalkyl, C2-c6 alkenyl, C2-C6 alkynyl, C1 6 perfluoroalkyl, F, Cl, Br, R9O-, R9S(o)m-, R~C(O)NR~, CN, NO2, (R8)2Nc(NR8)-~ R~C(O)-, R~OC(O)-, N3, -N(R8)2, R9OC(o)NR8- and Cl-C6 alkyl unsub,stituted or substituted by 1-3 groups .selected from: aryl, heterocyclyl, C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R8O-, R9S(o)m-~
R8C(O)NR8-, CN, (R8)2Nc(NR~ R8C(O)-, R~OC(O)-, N3, -N(R~)2 and R9OC(o)NR8-;
each R~ is independently selected from hydrogen, Cl-C6 alkyl, aryl and aralkyl;
each R9 is independently ~selected from Cl-C6 alkyl and aryl;
Al and A2 are independently selected from the group consisting of: a bond, -CH=CH-, -C_C-, -C(O)-, -C(O)NR8-, -NR8C(O)-, -O-, -N(R8)-, -s(o)2N(R~ -N(R~)s(o)2-~ and S(O)m;
V is selected from the group consisting of: hydrogen, heterocyclyl, aryl, Cl-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom ,selected from O, S, and N, and C2-c2o alkenyl, provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if Al isabond,n is0andA2isS(O)m;
provided that when V is heterocyclyl, attachment of V to R6 and to A
is through a substitutable ring carbon;
W represents heterocyclyl;
CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 each n and p independently represents 0, 1, 2t 3 or 4;
r i,s 0 to 5, provided that r is 0 when V is hydrogen, and tis 1.
DETAILED DESCRIPTION OF THE INVENTION
The compounds of this invention are useful in the inhibition of farne,syl-protein transferase and the farnesylation of the oncogene protein Ras, and thus are useful for the treatment of cancer.
The compound~s of the present invention may have asymmetric centers and occur as racemates, racemic mixtures and as individual diastereomers, with all possible isomers, including optical isomers, being included in the present invention.
When any variable (e.g. aryl, heterocycle, Rl, R2 etc.) occurs more than one time in any constituent, each definition is independent.
The term "alkyl" and the alkyl portion of alkoxy, aralkyl and similar terms, is intended to include branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, or 1-6 carbon atoms if unspecified. Cycloalkyl means 1-2 carbocyclic rings which are saturated and contain from 3-10 atom.s.
"Halogen" or "halo" as used herein means fluoro, chloro, bromo and iodo.
As used herein, "aryl" and the aryl portion of aralkyl, are intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 members in each ring, wherein at least one ring is aromatic. Examples of such aryl elements include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl. A preferred aralkyl group is benzyl.
The terms heterocyclyl, heterocycle and heterocyclic, as used herein, mean a 5- to 7-membered monocyclic or ~s- to 11-- membered bicyclic heterocyclic ring,s, either saturated or unsaturated, aromatic, partially aromatic or non-aromatic, and which con~sist of carbon atoms and from one to four heteroatoms selected from the group CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 con.sisting of N, O, and S. Thus, it includes any bicyclic group in which any of the above-defined heterocyclic rings i,s fu,sed to a benzene ring.
The ring or ring system may be attached at any heteroatom or carbon atom which re.sults in a stable structure. lt optionally contains 1-3 S carbonyl groups. Example,s of such heterocycles include, but are not limited to, azepinyl, benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, 10 dihydrobenzothiopyranyl sulfone, furyl, imidazolidinyl, imidazolinyl, imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl, i~sothiazolidinyl, isothiazolyl, isothiazolidinyl, morpholinyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 2-oxopyrrolidinyl, pyridyl, pyrazinyl, pyrazolidinyl, pyrazolyl, 15 pyridazinyl, pyrimidinyl, pyrrolidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiazolyl, thiazolinyl, thienofuryl, thienothienyl and thienyl.
"Heteroaryl" is a subset of heterocyclic as defined above, 20 and means a monocyclic or bicyclic ring system, with up to 7 members in each ring, wherein at least one ring is aromatic and wherein from one to four carbon atoms are replaced by heteroatom~s selected from the group consisting of N, O and S. Examples include benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, 25 benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, furyl, imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxadiazolyl, pyridyl, pyrazinyl, pyrazolyl, 30 pyridazinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiazolyl, - thienofuryl, thienothienyl and thienyl.
CA 0224961~ 1998-09-22 Lines drawn into ring sylstem.s from sub~stituents indicate that the bond may be attached to any of the substitutable ring carbon atoms.
Sub.stituted alkyl, substituted aryl, substituted heterocyclyl and substituted cycloalkyl mean alkyl, aryl, heterocyclyl and cycloalkyl groups, respectively, having from 1-3 substituents which are selected from: halo, aryl, heterocyclyl, C3 1() cycloalkyl, C2 6 alkenyl, C2 6 alkynyl, RXO-, R9S(o)m-~ RXC(O)NRx-, CN, (RX)2Nc(NRx) RXC(O)-, RXOC(O)-, N3, -N(Rx)2 and R~OC(O)NRX-. When for example, a substituted alkyl group i.s substituted with a "substituted aryl group", the aryl portion is substituted with 1-3 groups as defined above.
Preferably 1-2 groups are present on substituted alkyl, Isubstituted aryl, substituted heterocyclyl and substituted cycloalkyl, which are selected from: halo, aryl, R8O-, CN, RXC(O)- and -N(RX)2.
Preferably, Rla ,R1b and R2 are independently selected from: hydrogen, -N(R8)2, R8C(O)NR8- or unsubstituted or substituted C1-C6 alkyl wherein the substituent on the substituted Cl-C6 alkyl is selected from un.substituted or substituted aryl, -N(R~)2, RfsO-and R8C(o)NR8-Preferably, R3 and R4 are selected from: hydrogen, Cl-C6 alkyl, Br, Cl, F, R8O-, and CF3.
In a preferred group of compounds, A3 represents -C--C , -CR8=CR~s-, -C(O)- or a bond. A particularly preferred group of compounds within this subset includes compounds of formula I
~NHIBITORS OF FARNESYL-PROTEIN TRANSFERASE
S BACKGROUND OF THE ~VENTTON
The present invention relates to compounds which inhibit farnesyl protein transferase, a protein which is implicated in the oncogenic pathway mediated by Ras. The Ras proteins (Ha-Ras, Ki4a-Ras, Ki4b-Ra.s and N-Ras) are part of a signalling pathway that links cell surface growth factor receptors to nuclear signal.s initiating cellular proliferation. Biological and biochemical studies of Ras action indicate that Ras functions like a G-regulatory protein. In the inactive state, Ras is bound to GDP. Upon growth factor receptor activation Ras is induced to exchange GDP for GTP and undergoes a conformational change. The GTP-bound form of Ras propagates the growth stimulatory signal until the signal is termin~ted by the intrinsic GTPase activity of Ra,s, which returns the protein to its inactive GDP
bound form (D.R. Lowy and D.M. Willumsen, Ann. Rev. Biochem.
62:~51-~91 (1993)). Mutated ras genes (Ha-ras, Ki4a-)as, 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 ~re involved with Ras membrane localization, and all 3 modifications occur at the C-terminus of ~as. The Ras C-terminu.s contain,s a sequence motif termed a "CAAX" or "Cy.s-Aaal-Aaa2-Xaa"
box (Cys is cy~steine, Aaa is an aliphatic amino acid, the Xaa is any - 30 amino acid) (Willumsen et al., Natu) ~ 310:5~3-5~6 (19~4)). Depend-ing on the specific .sequence, this motif serves as a signal .sequence for - the enzymes farnesyl-protein transfera.se or geranylgeranyl-protein transferase, which catalyze the alkylation of the cysteine residue of the CAAX motif with a Cls or C2() isoprenoid, respectively. (S. Clarke., CA 0224961~ 1998-09-22 WO 97/36876 PCT/US97/~6257 Ann. Pev. Bi~chen~ 355-386 (1992); W.R. Schafer and J. Rine, Ann. Rel~. Genetics 30:209-237 (1992)). Ras proteins are known to undergo post-translational farnesylation. Other farnesylated proteins include the Ras-related GTP-binding proteins such as Rho, fungal 5 mating factors, the nuclear lamins, and the gamma subunit of transducin. James, et al., J. Biol. Chem. 269, 14182 (1994) have identified a peroxisome associated protein Pxf which is also farnesylated. James, et al., have also suggested that there are farnesylated proteins of unknown ~structure and function in addition 10 to those listed above.
lnhibition 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 demonstrated that certain inhibitors of farnesyl-protein transferase 15 selectively block the processing of the Ras oncoprotein intracellularly (N.E. Kohl et al., Science, 260:1934-1937 (1993) and G.L. James e~ al., Science, 260:1937-1942 (1993). Recently, it has been shown that an inhibitor of farnesyl-protein transferase blocks the growth of ~as-dependent tumors in nude mice (N.E. Kohl et al., Proc. Natl. Acad.
20 Sci U.S.A., 91:9141-9145 (1994) and induces regression of mammary and salivary carcinomas in ras transgenic mice (N.E. Kohl et al..
Nature Medicine, 1 :792-797 (1995).
Indirect inhibition of farnesyl-protein transfera~se in l'iVO
has been demonstrated with lovastatin (Merck & Co., Rahway, NJ) 25 and compactin (Hancock et al., ibid; Casey et al., ibid; Schafer et al., Science 245:379 (l9~S9)). The.se drugs inhibit HMG-CoA reductase, the rate limiting enzyme for the production of polyisoprenoids including farnesyl pyrophosphate. Farnesyl-protein transferase utilizes farnesyl pyrophosphate to covalently modify the Cys thiol group of the Ras 30 CAAX box with a farnesyl group (Reiss et al., Cell 62:~ g (1990);
Schaber et al., J. Biol. Chem., 2~5:14701-14704 (1990); Schafer et al., - Science, 249: 1133- 1139 (1990); Manne eF al., Pro( . Natl. Acad. Sci USA, 87:7541-7545 (1990)). Inhibition of farnesyl pyropho~phate biosynthesis by inhibiting HMG-CoA reductase blocks Ras membrane CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 Iocalization in cultured cells. However, direct inhibition of farnesyl-protein tran~sferase would be more specific, and thus preferable.
Inhibitor.s of farnesyl-protein transferase (FPTase) have been described in two general classes. The first are analogs of farnesyl 5 diphosphate (FPP), while the second class of inhibitors i~ related to the protein ~ubstrates (e.g., Ras) for the enzyme. The peptide derived 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., ihid; Reiss et. al., ihid; Reiss 0 et al., PNAS, 88:732-736 (1991)). Such inhibitors may inhibit protein prenylation while serving as alternate substrates for the farnesyl-protein tran,sferase 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)).
It has recently been reported that FPT-ase inhibitor,s also inhibit the proliferation of vascular smooth muscle cells and are therefore useful in the prevention and treatment of arteriosclerosi,s and diabetic disturbance of blood ves,sels (JP H7-112930).
It has recently been di,sclosed that certain tricyclic 20 compounds which optionally incorporate a piperidine moiety are inhibitors of FPTase (WO 95/10514, WO 95/1051~ and WO 95/10516).
Imidazole-containing inhibitors of farne~yl protein transferase have also been disclosed (WO 95/09001 and EP 0 675 112 Al).
The present invention addresses a compound of formula I:
V A1(CRla2) A2(CR'b2)n ~(W)~ (CR 2t~A - (CR 2)p or a pharmaceutically acceptable ~alt thereof, wherein:
CA 0224961~ 1998-09-22 W 097/36876 PCTrUS97/06257 Rla, R1b and R2 are independently selected from the group consisting of: hydrogen? aryl, substituted aryl, heterocyclyl, C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R~O-, R9S(o)m wherein m is 0, 1 or 2, R~C(O)NR8-, CN, NO2, (R8)2NC(NR8)-, R8C(O)-, R80C(O)-, N3, -N(R8)2, R90C(o)NR8- and Cl-C6 alkyl, unsubstituted or substituted by 1-3 groups .selected from the group consisting of: halo, aryl, heterocyclyl, C3-clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(o)m-~ R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R~)2 and 1 0 R9OC(o)NR8-;
R3 and R4 are independently selected from the group consisting of: H, F, Cl, Br, -NR82, CF3, NO2, R8O-, R9S(~)m-~
CF3(CH2)n-O-, R8C(O)NH-, H2NC(NH)-, R8C(O)-, R8OC(O)-, N3, CN, R9OC(o)NR8-, C1-C20 alkyl, substituted or unsubstituted aryl and ,substituted or unsubstituted heterocyclyl;
A3 is selected from: --C----C , --R8C=CR8---C(O)-, aryl, heteroaryl or a bond;
provided that when A3 is heteroaryl, attachment of A3 the remainder of the molecule is through substitutable heteroaryl ring carbon,s;
X represents aryl or heteroaryl;
provided that when X is heteroaryl, attachment of X the remainder of the molecule is through substitutable heteroaryl ring carbons;
R6 is independently selected from the group consisting of:
hydrogen, aryl, heterocyclyl, C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-6 perfluoroalkyl, F, Cl, Br, R~O-, R9S(o)m-~
R8C(O)NR8-, CN, NO2, (R8)2Nc(NR8)-~ R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(o)NR8- and Cl-C6 alkyl unsubstituted or substituted by 1-3 group~s selected from: aryl, heterocyclyl, C3-Clo cycloalkyl, CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/06257 C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R~O-, R9S(o)m-~ R8C(O)NR~-, CN, (R~)2NC(NR~)-, R~C(O)-, R~OC(O)-, N3, -N(R~)2 and R9OC(o)NR~-;
R7 is independently selected from the group consisting of:
hydrogen, aryl, heterocyclyl, C3-C1o cycloalkyl, C2-c6 alkenyl, C2-C6 alkynyl, C1 6 perfluoroalkyl, F, Cl, Br, R9O-, R9S(o)m-, R~C(O)NR~, CN, NO2, (R8)2Nc(NR8)-~ R~C(O)-, R~OC(O)-, N3, -N(R8)2, R9OC(o)NR8- and Cl-C6 alkyl unsub,stituted or substituted by 1-3 groups .selected from: aryl, heterocyclyl, C3-Clo cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R8O-, R9S(o)m-~
R8C(O)NR8-, CN, (R8)2Nc(NR~ R8C(O)-, R~OC(O)-, N3, -N(R~)2 and R9OC(o)NR8-;
each R~ is independently selected from hydrogen, Cl-C6 alkyl, aryl and aralkyl;
each R9 is independently ~selected from Cl-C6 alkyl and aryl;
Al and A2 are independently selected from the group consisting of: a bond, -CH=CH-, -C_C-, -C(O)-, -C(O)NR8-, -NR8C(O)-, -O-, -N(R8)-, -s(o)2N(R~ -N(R~)s(o)2-~ and S(O)m;
V is selected from the group consisting of: hydrogen, heterocyclyl, aryl, Cl-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom ,selected from O, S, and N, and C2-c2o alkenyl, provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if Al isabond,n is0andA2isS(O)m;
provided that when V is heterocyclyl, attachment of V to R6 and to A
is through a substitutable ring carbon;
W represents heterocyclyl;
CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 each n and p independently represents 0, 1, 2t 3 or 4;
r i,s 0 to 5, provided that r is 0 when V is hydrogen, and tis 1.
DETAILED DESCRIPTION OF THE INVENTION
The compounds of this invention are useful in the inhibition of farne,syl-protein transferase and the farnesylation of the oncogene protein Ras, and thus are useful for the treatment of cancer.
The compound~s of the present invention may have asymmetric centers and occur as racemates, racemic mixtures and as individual diastereomers, with all possible isomers, including optical isomers, being included in the present invention.
When any variable (e.g. aryl, heterocycle, Rl, R2 etc.) occurs more than one time in any constituent, each definition is independent.
The term "alkyl" and the alkyl portion of alkoxy, aralkyl and similar terms, is intended to include branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, or 1-6 carbon atoms if unspecified. Cycloalkyl means 1-2 carbocyclic rings which are saturated and contain from 3-10 atom.s.
"Halogen" or "halo" as used herein means fluoro, chloro, bromo and iodo.
As used herein, "aryl" and the aryl portion of aralkyl, are intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 members in each ring, wherein at least one ring is aromatic. Examples of such aryl elements include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl. A preferred aralkyl group is benzyl.
The terms heterocyclyl, heterocycle and heterocyclic, as used herein, mean a 5- to 7-membered monocyclic or ~s- to 11-- membered bicyclic heterocyclic ring,s, either saturated or unsaturated, aromatic, partially aromatic or non-aromatic, and which con~sist of carbon atoms and from one to four heteroatoms selected from the group CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 con.sisting of N, O, and S. Thus, it includes any bicyclic group in which any of the above-defined heterocyclic rings i,s fu,sed to a benzene ring.
The ring or ring system may be attached at any heteroatom or carbon atom which re.sults in a stable structure. lt optionally contains 1-3 S carbonyl groups. Example,s of such heterocycles include, but are not limited to, azepinyl, benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, 10 dihydrobenzothiopyranyl sulfone, furyl, imidazolidinyl, imidazolinyl, imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl, i~sothiazolidinyl, isothiazolyl, isothiazolidinyl, morpholinyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 2-oxopyrrolidinyl, pyridyl, pyrazinyl, pyrazolidinyl, pyrazolyl, 15 pyridazinyl, pyrimidinyl, pyrrolidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiazolyl, thiazolinyl, thienofuryl, thienothienyl and thienyl.
"Heteroaryl" is a subset of heterocyclic as defined above, 20 and means a monocyclic or bicyclic ring system, with up to 7 members in each ring, wherein at least one ring is aromatic and wherein from one to four carbon atoms are replaced by heteroatom~s selected from the group consisting of N, O and S. Examples include benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, 25 benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, furyl, imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxadiazolyl, pyridyl, pyrazinyl, pyrazolyl, 30 pyridazinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiazolyl, - thienofuryl, thienothienyl and thienyl.
CA 0224961~ 1998-09-22 Lines drawn into ring sylstem.s from sub~stituents indicate that the bond may be attached to any of the substitutable ring carbon atoms.
Sub.stituted alkyl, substituted aryl, substituted heterocyclyl and substituted cycloalkyl mean alkyl, aryl, heterocyclyl and cycloalkyl groups, respectively, having from 1-3 substituents which are selected from: halo, aryl, heterocyclyl, C3 1() cycloalkyl, C2 6 alkenyl, C2 6 alkynyl, RXO-, R9S(o)m-~ RXC(O)NRx-, CN, (RX)2Nc(NRx) RXC(O)-, RXOC(O)-, N3, -N(Rx)2 and R~OC(O)NRX-. When for example, a substituted alkyl group i.s substituted with a "substituted aryl group", the aryl portion is substituted with 1-3 groups as defined above.
Preferably 1-2 groups are present on substituted alkyl, Isubstituted aryl, substituted heterocyclyl and substituted cycloalkyl, which are selected from: halo, aryl, R8O-, CN, RXC(O)- and -N(RX)2.
Preferably, Rla ,R1b and R2 are independently selected from: hydrogen, -N(R8)2, R8C(O)NR8- or unsubstituted or substituted C1-C6 alkyl wherein the substituent on the substituted Cl-C6 alkyl is selected from un.substituted or substituted aryl, -N(R~)2, RfsO-and R8C(o)NR8-Preferably, R3 and R4 are selected from: hydrogen, Cl-C6 alkyl, Br, Cl, F, R8O-, and CF3.
In a preferred group of compounds, A3 represents -C--C , -CR8=CR~s-, -C(O)- or a bond. A particularly preferred group of compounds within this subset includes compounds of formula I
2~ wherein A3 represents -C--C-- or a bond.
Another preferred group of compounds includes the compounds of formula I wherein A3 represents aryl or heteroaryl.
Preferably R6 represents CN.
Preferably, R7represents hydrogen, unsubstituted or substituted Cl-C6 alkyl.
Preferably, RX represent.s H or Cl ~ alkyl, and R9 i.s C
- alkyl.
Preferably, Al and A2 are independently selected from:
CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 a bond, -C(O)NRX-, -NR~C(O)-, -O-, -N(RX)-, -S(O)2N(RX)- and-N(Rg)S(0)2--- Preferably, V is selected from hydrogen, heterocyclyl and aryl. More preferably V is phenyl.
Preferably, W is heterocyclyl selected from imidazolinyl, imidazolyl, oxazolyl, pyrazolyl, pyyrolidinyl, thiazolyl and pyridyl.
More preferably, W i.s selected from imidazolyl and pyridyl.
Preferably X represents aryl. In particular, X can represent phenyl.
Preferably, m is 0 or 2.
Preferably n and p are 0, 1, 2 or 3.
A subset of compounds of the invention is represented by forrnula Ia:
(R )r V - A1 (C R 1 a2)nA2(C R 1 b2)~3 N ~R3 (CR22)p--A3- (CR22)p X
\ 4 15 wherein:
R3, R4, A3, R~, R9, X, m, n, p and r are as originally defined;
each R 1 a and R2 is independently selected from hydrogen and Cl -C6 alkyl;
each Rlb is independently selected from: hydrogen, aryl, heterocyclyl, C3 1() cycloalkyl, C2-~ alkenyl, R~O-, -N(R8)2 and Cl-C6 alkyl unsubstituted or substituted by aryl, heterocyclyl, cycloalkyl, alkenyl, RgO- and -N(RX)2;
CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 R6 is independently selected from: hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-c6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl, R80-, R~C(O)NR~-, CN, NO2, (R~s)2N-c(NR~ R~sC(O)-, R~OC(O)-, -N(Rg)2, or R9OC(o)NR8-, and Cl-C6 alkyl substituted by Cl-C6 5 perfluoroalkyl, R80-, R8C(O)NR8-, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2 and R9OC(o)NR8-;
R7 repre.sents H or Cl-6 alkyl;
A 1 and A2 are independently selected from: a bond, -CH=CH-, -C_C-, -C(O)-, -C(O)NR2~-, O, -N(R8)- and S(O)m;
and V is selected from: hydrogen; aryl; heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, ~uinolinyl, isoquinolinyl and thienyl; Cl-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if Al is a bond and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R6 and to Al is through a substitutable ring carbon.
A second subset of compounds of the present invention is represented by formula Ib:
(R6) /~7~ / R3 V-A1(CR1a2)nA2(CR1b2)n-\W~-(CR22)p-R3C=CR8~CR22),~X
Ib wherein:
Rla Rlb, R2, Al, A2, R3, R4, R6, RX, R9, X, m, n, p and r are as originally defined;
R7 is selected from: hydrogen and Cl-C6 alkyl;
CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/06257 V is selected from: hydrogen, heterocyclyl ~elected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl and thienyl; aryl; Cl-c2o alkyl wherein from 0 to 4 carbon atom.s are replaced with a heteroatom selected from O, S, and N, S and C2-C20 alkenyl, provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if A 1 is a bond, n i~s 0 and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R~ and to Al is 10 through a substitutable ring carbon;
and W represents heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl and isoquinolinyl.
A third subset of compounds of the present invention is repre~ented by formula Ic:
(R6) (17~ ~R3 V-A1(CR1a2)nA2(CR1b2)n\W~~(CR22)pC_C--(CR2z~X
Ic R4 wherein:
Rl~ Rlb R2, Al, A2, R3, R4, R6, RX, R9, X, m, n, p and r are as originally defined;
R7 is selected from: hydrogen and Cl-C6 alkyl;
V is selected from: hydrogen, heterocyclyl ,selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, aryl, Cl-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, CA 0224961~ 1998-09-22 provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if Al i~sabond,n isOandA2isS(O)m;
provided that when V is heterocycle, attachment of V to R~ and to Al is 5 through a substitutable ring carbon;
and W represents heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl and i.soquinolinyl.
A fourth embodiment of the invention is described in accordance with formula Id:
H
)~N
,=~N~\J ~3 ~¦~ (CR 2)p-A - (CR 2)p X~
n Id wherein:
each R2 is independently selected from hydrogen and Cl-C6 alkyl;
R3, R4, A3, RX, R9, X, m and p are as originally defined;
and R6 is selected from the group consisting of: hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl, R~O-, R8C(O)NR8-, CN, NO2, (R8)2N-C(NR8)-, R8C(O)-, 25 R8OC(O)-, -N(R8)2, or R9OC(o)NR~- and Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl, R80-, R8C(O)NR~-, (R8)2N-C(NR8)-, R8C(O), R8OC(O)-, -N(R~)2 or R9OC(o)NRX-.
W O 97/36876 PCT~US97/06257 A fifth sub.set of compounds of the invention is represented by formula le:
H
)~N
(C R22)p A3 - (C R22)--X
NC
le wherein:
X and A3 are as originally defined;
each R2 is independently selected from: hydrogen and Cl-C6 alkyl;
R3 and R4 are independently ,selected from H, F, Cl, Br, N(R8)2, CF3, N02, (R8)o-, (R9)S(o)m-, (R8)C(O)NH-, H2N-C(NH)-, (R8)C(O)-, (R8)0C(O)-, N3, CN, (R9)OC(O)NR8-, Cl-C20 alkyl, substituted or unsub.stituted aryl, and substituted or unsubstituted heterocyclyl;
and R~, R9, m and p are a.s originally defined.
Specific examples of compound.s of the invention are:
NC~ C--C~
<'~
N
PCTrUS97/06257 NC ~ CH--CH~
N
NC ~ C--C
N
~\N ~ C H = C H ~ ~
NHo Cl NC ~N~C _C
N
NC~3~N
NC ~ N~
N
~>~N~? CH2-C--C
N
NC ~C--C
~\N~
S N
~_N CN
N
OH
N
' ~
N c'(3J ~3 CA 0224961~ 1998-09-22 and the pharmaceutically acceptable salts and isomers thereof.
The pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds 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, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.
The pharmaceutically acceptable salts of the compounds of this invention can be synthesized from the compounds of this invention which contain a basic moiety by conventional chemical methods. Generally, the salts are prepared either by ion exchange chromatography or by reacting the free base with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid in a suitable ,solvent or variou,s combinations of solvents.
Reactions used to generate the compounds of this invention are prepared by employing reactions as shown in the Schemes, 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. Substituents R and R CH2-, as shown in the Schemes, represent the substituents R8, R9 and others, depending on the compound of the instant invention that is being synthesized. The variable p' represents p-l.
These reactions may be employed in a linear sequence to provide the compounds of the invention or they may be u,sed to synthesize fragments which are subsequently joined by the alkylation reactions described in the Schemes.
Svnopsis of Schemes CA 0224961~ 1998-09-22 The requisite intermediates are in ~some cases commercially available, or can be prepared according to literature procedures.
Schemes 1-2 illustrate the synthesi.s of one of the preferred embod~-ment.s of the instant invention, wherein the variable W is present as an imidazolyl moiety that is sub,stituted with a suitably sub,stituted benzyl group. Substituted protected imidazoles can be prepared by method.s such a,s those de.scribed by F. Schneider, Z. Physiol. Chem., 3:206-210 (1961) and C.P. Stewart, Biochem. Journal, 17:130-133(1923).
Benzylation and deprotection of the imidazole alkanol provide.s intermediate III which can be oxidized to the corresponding aldehyde IV. Also, while X is shown as a phenyl ring, other aryl and heteroaryl groups can be substituted therein without departing from the invention.
The a~dehyde whose synthesis is illustrated in Scheme 1 may be reacted with ~ suitably substituted aralkyne, to provide the intermediate compound V. Compound V can be selectively hydrogenated across the unsaturated bond under standard conditions, such as those illustrated, to provide Compound VI.
Schemes 3- 10 illustrate syntheses of suitably substituted aldehydes u,seful in the syntheses of the instant compounds wherein the variable W is a pyridyl moiety. Similar synthetic strategies for preparing alkanols that incorporate other heterocyclic moieties for variable W can be discerned from the teachings herein.
Generally the aldehyde i,s reacted with an appropriately substituted aralkyne using n-BuLi, after which the triple bond can be reduced. As shown in Schemes 2, 4, 6 and ~s reduction of the alkyne triple bond using Pd/BaSO4 Iproduces the Z-olefin isomer almost exclusively. By substituting sodium bi,~(2-methoxyethoxy)aluminum hydride (RED-AL) in toluene, one can readily obtain the E-allylic alcohol from propargylic alkynes used in the present invention.
In the preparation methods described herein, reactive groups may remain blocked until the final product is prepared, - essentially in protected form, after which a final deprotection step is conducted. These blocking groups are readily removable i.e., they can be removed, if desired, by procedures which will not cause cleavage or CA 0224961~ 1998-09-22 other disruption of the remaining portions of the molecule. Such procedures include chemical and enzymatic hydrolysis, treatment with chemical reducing or oxidizing agents under mild conditions, treatment with fluoride ion, treatment with a transition metal catalyst and a 5 nucleophile and catalytic hydrogenation.
Examples of suitable hydroxyl protecting groups are:
t-butylmethoxyphenylsilyl, t-butoxydiphenylsilyl, trimethylsilyl, triethylsilyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, benzyloxycarbonyl, t-butyloxycarbonyl, 2,2,2-trichloroethyloxy-10 carbonyl and allyloxycarbonyl. Preferred hydroxyl protecting groupsare trimethylsilyl and triethylsilyl.
Examples of suitable carboxyl protecting groups are:
benzhydryl, o-nitrobenzyl, p-nitrobenzyl, 2-naphthylmethyl, allyl, 2-chloroallyl, benzyl, 2,2,2-trichloroethyl, trimethylsilyl, 15 t-butyldimethylsilyl, t-butyldiphenylsilyl, 2-(trimethylsilyl)ethyl, phenacyl, p-methoxybenzyl, acetonyl, p-methoxyphenyl, 4-pyridylmethyl and t-butyl. A preferred-carboxyl protecting group is p-nitrobenzyl.
Many other suitable hydroxyl and carboxyl protecting 20 groups are known in the art. See, e.g., T.W. Greene, Protective Groups in Or~anic Synthesis, John Wiley & Sons, Inc., 1981 (Chapters 2 and 5).
W O 97/36876 PCT~US97/06257 ~(cR22)Pl-cH2oH Prot1X Et3N Prot 'N~/~>
H DMF
lla rBr (C R22)p -C H20Ac R 6~EtOAc AC20, Py ,=1=~
Prot1~N~ N 2. N-deprotect CR2 ) -CH OAC N~(CR 2)p-CH20H
N~ LiOH
R6/~ THF, H20 ~
N,~(CR 2)p,-CHO HO r/R3 ~N ~/ (CR22)p.-CH-C_C~_~
SO3Py, Et3N ~ n-BuLi, THF ~ > R4 DMSO~ H
V
W O 97/36876 PCT~US97/06257 )p.-CH-C_C{;~
~N R4 R6 H2, Pd/C
V
MeOH, EtOAc H2, Pd/BaSO4 MeOH, EtOAc HO R3 HO 3,~cR22)p-cH
~$CR22)p~-cH-CH=c~_;~ N
~N R4 (predominantly R6/~
~, Z isomer) Vlb R6/~
Vla CH3 1) HN~2~Br2 CO2CH3 ~ 2) KMnO
H2N N 3) MeOH,H+ Br N
r ~\MgCI R6 ~ ~,C02CH3 ZnCI2,NiCI2(Ph3P)2 N
NaBH4 (excess) ~,CH20H
SO3 Py Et3N ~CHO
n-BuLi,THF R6 H--C--C{ ~ ~)~CH
W O 97/36876 PCTrUS97/06257 H2, Pd/BaSO4 MeOH, EtOAc H2, Pd/C
~" MeOH, EtOAc R6 ~CH-CH=CH~
(predominantlyZ isomer) ~ ~CH~ R4 W O 97136876 PCTrUS97/06257 1. EtOC(=O)CI
~,CO2CH3 ~Z\ 9C CN ~C02CH3 N 3. S, xylene, heat N
NaBH4 ~ SO3Py, Et3N ~
(excess) ~"CH20H DMSO ~CHO
Br CO2CH3[~\ MgCI [~j ZnCI2, Nicl2(ph3p)2 ~CO2CH3 NaBH4 ¦ SO3Py, Et3N
~CH20H ~ ~CHO
(excess) N DMSO NJJ
N-BuLi, THF ll f, OH
H = ~R~
OH
~,CH
N
H2, Pd/BaSO4 H2, Pd/C
MeOH, EtOAc MeOH, EtOAc ¢~3,CH-CH=CH~ 3,OH
(predominantly Z isomer) Br~ 1. LDA, CO2 Br~
N 2. MeOH, H+ N
r ZnC12, NiC12(Ph3P)2 N
NaBH4 (excess) ~ ( ~20H S03Py, Et3N
DMSO
R~
N
H2, Pd/BaSO4 H2, Pd/C
MeOH, EtOAc MeOH, EtOAc ~' N N
(predominantly Z isomer) W O 97/36876 PCTrUS97/06257 1. LDA, CO2 [~Br 2. (CH3)3sicHN2 R6 ~Br R6 ~
~, co2CH3 Zn, NiC12(Ph3P)2 N~
R6 ~
excess NaBH4 ~ SO3 Py, Et3N
N ~CH20H DMSO
~, R6 ~ R6 ~ OH
~CHO n-BuLi, THF N~ h R3 -W O 97t36876 PCTrUS97106257 R6 ~q OH
~- = 7 H2, Pd/C R4 MeOH, EtOAc R6 ~
~/
BocN H ~
BocNH CHO
n-BuLi, THF
BocN H
/=~ R CF3CO2H
BocNH~ ~ R4 CH2CI2 OH
R3 Boc20 NH2/~ R4 CH2CI2 OH
BocNH (R )r~ CHO
NH2~ R4 NaBH(OAc)3 OH Et3N, CICH2CH2CI
BocN H
(R )r~--CH2 ~ ~:~R3 W O 97/36876 PCT~US97/06257 BocN H
~ R CF3CO2H
(R )~ CH 'NH~ R4 NaHCO3 NH~ ~R
(R )r~--CH2 ~ R4 AgCN
(R )r~ CH/ \~ 4 H ~ R3 Me><O~ ~
Me N~\cHo R4 Boc n-BuLi, TH F
>< ~ R3 t-Bu(Me)2Si-CI
, ~R4 imidazole, DMF
Boc OH
Me>< ~ R3 TsOH, H20 Boc R benzene O-SiMe2(t-Bu) HO
~ R CICOCOCI
BocNH~ R4 DMSO CH2C12 O-SiMe2(t-Bu) (c2H5)3N
O H
R3 1. R'MgX
BocNH--~R4 2. TFA
O-SiMe2(t-Bu) CH2CI2 HO R' ~ ~/ R
H2N~ R4 OH
HO
~ R CF3CO2H
BocNH~ R4 CH2C12 O-SiMe2(t-Bu) HO
~ R3 R'CHO
H2N~ R4 NaBH(OAc)3 HO
R'CH2~N A,~ R4 OH
HO 3 N=~ ~= N
=~ R ~ ' S ~
BocNH~ ~ R4 ~2 ,, O-SiMe2(t-Bu) NaH, DMF 0~C
Boc--N1 ~=~ R3 R'SH
4 (C2H5)3N ~, R CH3oH
O-SiMe2(t-Bu) R'S
~ R CF3CO2H
BocNH '\~ R4 CH2C12 O-SiMe2(t-Bu) R'S ~ R3 H2N~ R4 OH
W O 97/36876 PCTrUS97/06257 HO,~1) Boc20, K2C~3 HO~
~/ THF-H20 2) CH2N2, EtOAc J~
H2NC02H BocNH CO2CH3 HO,,~
LiAlH4 ,b~l R'CH2X
THF l Cs2CO3 0-20~C BocNH CH2OH DMF
R'C H20 R'C H20 )~ pyridine SO
DMSO
BocNH CH20H (c2H5)3N BocNH~CHO
WO 97136876 PCTnUS97/062~7 SCHEME 16 (cont.) R'CH ;~
BocNH CHO n-BuLi, THF
R'CH20~
BocNH J~ _ lc/R3 HCI / \ BF3-0Et2, EtSH
ETOAc / \ CH2CI2 R'C H20~3 HO~
H2NJ~ ~R H2NJ~
The instant compound.s are useful in the treatment of cancer. Cancers which may be treated with the compounds of this 5 invention include, but are not limited to, colorectal carcinoma, exocrine pancreatic carcinoma, myeloid leukemia~ and neurological tumor.s. Such tumors may arise by mutations in the ra~ genes CA 0224961~ 1998-09-22 W O 97136876 PCTrUS97106257 themselves, mutations in the proteins that can regulate Ras activity (i.e., neurofibromin (NF-l), neu, scr, abl, lck, fyn) or by other mechani~sms.
The compounds of the instant invention inhibit farnesyl-protein transferase and farnesylation of the oncogene protein Ras.
5 The instant compounds may also inhibit tumor angiogenesis, thereby affecting the growth of tumors (J. Rak et al. Cancer Researc~h, 55:4575-4580 (1995)). Such anti-angiogenic 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 diseases where Ras proteins are aberrantly activated a,s a result of oncogenic mutation in other genes (i.e., the Ras gene itself i.s 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 component of NF-l is a benign proliferative disorder.
The instant compounds may also be useful in the treatment of 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 the prevention of restenosis after percutaneous transluminal coronary angioplasty by inhibiting neointimal formation (C. Indolfi et al. Natu~ e medicine, I :541 -545(1995).
The instant compounds may also be useful in the treatment and prevention of polycystic kidney disease (D.L. Schaffner et al.
Ame~ican Journal of Pathology, 142:1051-1060 (1993) and B. Cowley, Jr. et al.FASFB Journal, 2:A3160 (19~s~)).
The instant compounds may also be useful for the treatment of fungal infections.
The compounds of this invention may be administered to m~mm~ls, preferably humans, either alone or, preferably, in combina-- tion with pharmaceutically acceptable carriers or diluents, in the form of a pharmaceutical composition, which is comprised of a compound of formula I in combination with a pharmaceutically acceptable carrier.
CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/06257 The compounds can be administered orally, topically, rectally, vaginally transderrnally or parenterally, including the intravenous, intramuscular, intraperitoneal and ,subcutaneous routes of administration.
For oral use, the compound is administered, for example, 5 in the form of tablets or capsules, or as a solution or .suspen,sion. In the case of tablets for oral use, carriers which are commonly used include lactose and corn ,starch; lubricating agents, such as magnesium stearate, are commonly added. For oral administration in capsule form, diluents also include lactose and dried corn starch. When a4ueous suspensions 10 are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If de,sired, certain sweetening and/or flavoring agents may be added. For intramuscular, intraperi-toneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared, the pH of the solution is suitably 15 adjusted and the product is buffered. For intravenous use, the total concentration is controlled to render the preparation substantially isotonic.
As used herein, the term "composition" i.s intended to encompass a product comprising the specified ingredients in the specific 20 amounts, as well as any product which results, directly or indirectly, from combination of the specific ingredients in the specified amount.s.
The compound,s of the instant invention may also be co-~lmini~tered in therapeutic composition~s that also contain other well known therapeutic agents that are selected for their particular 25 usefulness against the condition that is being treated. For example, the instant compound.s may be useful in combination with known anti-cancer and cytotoxic agents. Similarly, the instant compounds may be u.seful in combination with agents that are effective in the treatment and prevention of NF-I, restinosis, polycystic kidney disease, infection.s of 30 hepatitis delta and related viruses and fungal infections.
If formulated a~s a fixed dose, such combination products - employ a compound of this invention substantially within the dosage range described below and other pharmaceutically active agent(s~
typically within the acceptable dosage range. Compounds of the CA 0224961~ 1998-09-22 W O 97t36876 PCTrUS97/06257 -3~-instant invention may alternatively be used sequentially with known pharmaceutically acceptable agent(s) when a combination formulation i.s inappropriate.
The daily dosage will normally be determined by the 5 prescribing physician, who may vary the dosage according to the age, weight, and response of the individual patient, a.s well as the severity of the patient's condition.
In one exemplary application, a suitable amount of compound is administered to a m~mm~l undergoing treatment for 10 cancer. Adrnini.stration occurs in an amount between about 0.1 mg/kg of body weight to about 60 mg/kg of body weight per day, preferably of between 0.5 mg/l~g of body weight to about 40 mgtkg of body weight per day.
The compounds of the instant invention are also useful 15 as a component in an assay to rapidly determine the presence and quantity of farnesyl-protein transferase (FPTase) in a composition.
Thus the composition to be tested may be divided and the two portions contacted with mixtures which comprise a known substrate of FPTase (for example a tetrapeptide having a cysteine at the amine 20 terminus) 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 farnesylate the substrate, the chemical content of the assay mixture.s may be determined by well known immuno-25 logical, radiochemical orchromatographic techniques. Becausethe 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 compound of the instant invention relative to the presence of the unchanged sub~strate in the a,ssay 30 containing the instant compound i.s 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 u~seful in identifying tissue samples which contain farnesyl-protein transferase and quanti-CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/062~7 tating the enzyme. Thus, potent inhibitor compounds of the instant invention may be u~sed in an active site titration assay to determine the quantity of enzyme in the sample. A serie~s of .samples composed of aliquots of a tissue extract containing an unknown amount of farne,syl-5 protein transferase, an excess amount of a known substrate of FPTa~se (for example a 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 10 that has a Ki Isubstantially 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.
(+)-1 -(4-CYANOBENZYL)-S-1 ( I -HYDROXY-3-PHENYL)-2-PROPYNYLlIMIDAZOLE HYDROCHLORIDE
20 Step A: Preparation of l-triphenylmethyl-4-(hydroxymethyl)imidazole To a solution of 4-(hydroxymethyl)imidazole hydrochloride (35.0 g, 260 mmol) in 250 mL of dry DMF at room temperature was added triethylamine (90.6 mL, 650 mmol). A white 25 solid precipitated from the solution. Chlorotriphenylmethane (76.1 g, 273 mmol) in 500 mL of DMF was added dropwise. The reaction mixture was .stirred for 20 hour.s, poured over ice, filtered, and washed with ice water. The resulting product wa,s slurried with cold dioxane, filtered, and dried in vacuo to provide the titled product as a white solid 30 which was sufficiently pure for use in the next ,step.
Step B: Preparation of l-triphenylmethyl-4-(acetoxymethyl)imidazole CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 Alcohol from Step A (260 mmol, prepared above) was suspended in 500 mL of pyridine. Acetic anhydride (74 mL, 7~0 mmol) wa.s added dropwi.se, and the reaction was stirred for 4~ hours during which it became homogeneous. The solution was poured into 2 S L of EtOAc, washed with water (3 x 1 L), 5% aq. HCI soln. (2 x 1 L), sat. aq. NaHCO3, and brine, then dried (Na2SO4), filtered, and concentrated in vacuo to provide the crude product. The acetate was isolated a~s a white powder (85.X g, 86% yield for two ~teps) which wa~s sufficiently pure for use in the next reaction.
Step C: Preparation of 1-(4-cyanobenzyl)-S-(acetoxymethyl)imidazole hydrobromide A solution of the product from Step B (85.8 g, 225 mmol) and ~-bromo-p-tolunitrile (50.1 g, 232 mmol) in 500 mL of EtOAc was lS stirred at 60 ~C for 20 hours, during which a pale yellow precipitate formed. The reaction wa.s cooled to room temperature and filtered to provide the solid imidazolium bromide salt. The filtrate was concen-trated in vacuo to a volume 200 mL, reheated at 60~C for two hours, cooled to room temperature, and filtered again. The filtrate was 20 concentrated in vacuo to a volume 100 mL, reheated at 60~C for another two hours, cooled to room temperature, and concentrated in vacuo to provide a pale yellow solid. All of the solid material was combined, dissolved in 500 mL of methanol, and warmed to 60~C.
After two hours, the solution was reconcentrated in V~lCUo to provide 25 a white solid which was triturated with hexane to remove soluble materials. Removal of residual solvents in vacuo provided the titled product hydrobromide a~s a white solid (50.4 g, 67% yield, ~9% purity by HPLC) which was used in the next step without further purification.
30 Step D: Preparation of 1-(4-cyanobenzyl)-S-(hydroxymethyl)imidazole - To a solution of the acetate from Step C (50.4 g, 150 mmol) in l .S L of 3: I THF/water at 0 ~C was added lithium hydroxide monohydrate (18.9 g, 450 mmol). After one hour, the reaction was CA 0224961~ 1998-09-22 concentrated in vacuo, diluted with EtOAc (3 L), and washed with water, .sat. a~. NaHCO~ and brine. The solution was then dried (Na2SO4), filtered, and concentrated in vacuo to provide the crude product (26.2 g, ~2% yield) as a pale yellow fluffy solid which was ~S sufficiently pure for use in the next ~itep without further purification.
Step E: Preparation of 1-(4-cyanobenzyl)-~S-imidazolecarboxaldehyde To a solution of the alcohol from Step D (21.5 g, 101 mmol) in 500 mL of DMSO at room temperature was added triethylamine (56 mL, 402 mmol), then SO3-pyridine complex (40.5 g, 254 mmol). After 45 minutes, the reaction was poured into 2.5 L
of EtOAc, washed with water (4 x 1 L) and brine, dried (Na2SO4), filtered, and concentrated in vacuo to provide the aldehyde (1~.7 g, 1~ ~8% yield) as a white powder which was sufficiently pure for use in the next step without further purification.
Step F: Preparation of (+)-1-(4-cyanobenzyl)-5-[(1-hydroxy-3-phenyl)-2-propynyllimidazole hydrochloride To a solution of phenylacetylene (0.172 mL, 1.56 mmol) in 5 mL of THF at 0 ~C was added n-butyllithium (0.530 mL, 2.5 M in hexanes, 1.32 mmol). After 15 minutes, the aldehyde from Step E (254 mg, 1.20 mmol) was added, and the reaction was stirred for 30 minutes.
The reaction was quenched with sat. aq. NaHCO3, poured into EtOAc, washed with sat. aq. NaHCO3 and brine, dried (Na2SO4), filtered, and concentrated in vacuo. The resulting product was purified by silica gel chromatography (35-50% acetone/CH2CI2) to provide 1 ~7 mg of the desired alcohol. A portion of this was taken up in CH2CI2 and treated with excess 1 M HCI/ether solution, and concentrated in vacuo. The titled product hydrochloride wa.s isolated as a white ~olid.
- FAB mas.~i spectrum m/e 314 (M+l).
Analysis calculated for C2~HI~N3O ~ 1.0 HCI - 0.90 H2O:
C, 65.63; H, 4.90; N, 11.4~;
W O 97/36876 PCTrUS97/06257 Found: C, 65.81; H, 4.98; N, 1 1.17.
(+)-1 -(4-CYANOBENZYL)-5-~( 1 -HYDROXY-4-PHENYL)-3-BUTYNYLlIMIDAZOLE HYDROCHLORIDE
To a solution of t-butyllithium in 1.5 mL of THF (0.78 mL
of 1.7 M in pentane, 1.32 mmol) at -78~C was added tetramethylethyl-enediamine (0.199 mL, 1.32 mmol) and 1-phenyl-1-propyne (0.150 mL, 1.20 mmol). The solution was warmed to 0 ~C for one hour, then cooled to -78 ~C. The aldehyde from Step E of Example 1 (225 mg, 1.07 mmol) on 1.0 mL THF was added, and the reaction allowed to warm to 0 ~C. After 30 minutes, the reaction was quenched with sat.
aq. NaHCO3, poured into EtOAc, washed with sat. aq. NaHCO3 and brine, dried (Na2SO4), filtered, and concentrated in vacuo. The product was purified by silica gel chromatography (3-4% MeOH/CH2CI2), then taken up in CH2CI2 and treated with excess 1 M HCI/ether solution, and concentrated in vacuo to provide the titled product hydrochloride (48 mg) as a pale yellow foam.
FAB mass spectrum m/e 32~ (M+l).
Analysis calculated for C2lH17N3O ~ 1.10 HCl ~ 0.10 Et2O:
C, 68.56; H, 5.14; N, 11.21;
Found: C, 68.30; H, 5.04; N, 11.06.
(+)-3-(4-CYANOBENZYL)-4-l ( 1 -HYDROXY-3-PHENYL)-2-PROPYNYLlPYRIDINE HYDROCHLORIDE
- Step A: Preparation of 3-(4-cyanobenzyl)pyridin-4-carboxylic acid methyl ester CA 0224961~ 1998-09-22 W 097/36876 PCTrUS97/06257 A solution of 4-cyanobenzyl bromide (625 mg, 3.27 mmol) in dry THF (4mL) was added slowly over~3 min. to a suspension of activated Zn (dust; 250 mg) in dry THF (2 mL) at O~ under an argon atmosphere. The ice-bath was removed and the slurry was stirred at S room temperature for a further 30 min. Then 3-bromopyridin-4-carboxylic acid methyl ester (~40 mg. 2.5 mmol) followed by dichlorobis(triphenylphosphine)nickel (Il) (50 mg). The resultant reddish-brown mixture was stirred for 3h at ~40-45~C. The mixture was cooled and distributed between EtOAc (l00 ml) and 5% aqueou~
citric acid (50 mL). The organic layer was washed with H2O (2X50 mL), dried with Na2SO4. After evaporation of the solvent the residue was purified on silica gel, eluting with 35% EtOAc in hexane to give 420 mg as a clear gum. FAB ms (M+l) 253.
Step B: Preparation of 3-(4-cyanobenzyl)-4-(hydroxymethyl)pyridine The title compound was obtained by sodium borohydride (300 mg) reduction of the ester from Step A (415 mg) in methanol (5 mL) at room temperature. After stirring for 4 h the solution was evaporated and the product was purified on silica gel, eluting with 2%
methanol in chloroform to give the title compound. FAB ms (M+l ) 225.
Step C: Preparation of 3-(4-cyanobenzyl)-4-pyridinal The title compound was obtained by activated manganese dioxide (1.0g) oxidation of the alcohol from Step B (240 mg, 1.07 mmol) in dioxane (10 mL) at reflux for 30 min. Filtration and evaporation of the solvent provided title compound, mp ~0-~s3OC.
Step D: Preparation of (+)-3-(4-cyanobenzyl)-4-[(1-hydroxy-3-phenyl)-2-propynyllpyridine hydrochloride The titled compound is prepared from the pyridinal from Step C using the procedured in Step F of Example 1. The product is purified by silica gel chromatography, then taken up in CH2CI2 and CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 treated with excess 1 M HCl/ether solution, and concentrated i~ cuo to provide the titled product hydrochloride.
1 -(4-BIPHENYLMETHYL)-5-(4-CYANOBENZYL)IMIDAZOLE
HYDROCHLORIDE SALT
Step A: 1 -Trityl-4-(4-Cyanobenzyl)-imidazole.
To a suspension of activated zinc dust (3.57g, 54.98mmol) in THF (50ml)was added dibromoethane (0.315m1, 3.60mmol) and the reaction stirred under argon at 20~C. The suspension was cooled to 0~C
and (x bromo-p-toluinitrile (9.33g, 47.6mmol) in THF (lOOml) was added dropwise over a period of 10 min. The reaction was then allowed to stir at 20~C for 6hr and bi,s(triphenylphosphine)Nickel II chloride (2.4g, 3.64mmol) and 5 -iodotrityl imidazole (15.95g, 36.6mmol) was added in one portion.The resulting mixture was stirred 16hr at 20~C
and then quenched by addition of saturated NH4CI solution (lOOml) and the mixture stirred for 2 hours. Saturated NaHCO3 solution was added to give a pH of P~ and the solution wa,s extracted with EtOAc (2x250ml), dried MgSO4 and the solvent evaporated in vacuo. The residue was chromatographed (sio2~ 0-20% EtOAc/CH2Cl2 to afford the title compound as a white solid.
1H NMR ~ CDC13 (7.54 (2H, d, J=7.9Hz), 7.38(1H, s), 7.36-7.29 (1 lH, m), 7.15-7.09(6H, m), 6.58(1H, s), and 3.93(2H, s)ppm.
Step B: 1 -(4-Biphenylmethyl)-5-(4-cyanobenzyl)imidazole hydochloride salt To 1-Trityl-4-(4-Cyanobenzyl)-imidazole (60~.~ mg, 1.43 mmol) in acetonitrile (2 ml) was added 4-chloromethyl biphenyl (290mg, 1.43 mmol) and the mixture heated at 55~C for 16 hours. The - residue was dissolved in methanol (30 ml) and heated at reflux for 20 mins, cooled and evaporated to dryness. The residue was partitioned between saturated NaHCO3 solution and CH2C12. The organic layer was dried (MgSO4) and the solvent evaporated in vacuo. The re~idue wa~ chromatographed (SiO2, 5% methanol in CH2C12) to afford the imidazole which was converted to the HCI salt by treatment with one equivalent of HCI in a4ueous acetonitrile. Evaporation of Isolvent in 5 vacuo afforded the title compound as a white powder.
Anal. Calcd for C24HlgN3- 1.00 HCI:
C, 74.70; H, 5.22; N, 10.~9.
Found: C, 74.70; H, 5.31; N, 10.77.
FAB MS 350 (MH+) 10 lH NMR CD30D ~ 9.03(1H, s), 7.65-7.50(5H, m), 7.44(2H, t, J=7.5Hz), 7.39(1H, s), 7.35(1H, t, J=7.3Hz), 7.26(2H, d, J=~.lHz), 7.20(2H, d, J=X.lHz), 5.42(2H, s), and 4.17(2H, ,s) ppm.
I l -(4-CYANOBENZYL)IMIDAZOL-5-YLl(~ 1.1 '-BIPHENYLl-4-YL)METHANOL
A Grignard reagent, freshly prepared from 4-bromo[l,l'-20 biphenyl] (116 mg, 500 ,umol) and magnesium turnings (1~ mg, 730 ,umol) in dry THF (500 ,ul) was added to a dry Argon-purged 3mL flask containing the aldehyde (105 mg, 500 }lmol) in dry THF (200 ~L) with vigorous stirring at room temperature. After I hour the reaction was quenched with sat. NH4CI (5 mL) and distributed between EtOAc (50 25 mL) and H2O (50 mL). The organic phase was evaporated and the residue was chromatographed on silica gel (CHCI3-MeOH (20:1)) to yield title (117 mg).
FAB ms (M+l) 366.25.
Anal. Calc. for C24H IgN3O-0.10 CHCI3-0.10 CH3OH;
C, 76.37: H, 5.16: N, 11.04.
Found: C, 76.13; H, 5:10; N, 10.76.
W O 97/36876 PCTrUS97/06257 11-(4-CYANOBENZYL)IMIDAZOL-5-YLl(l l.l'-BIPHENYLl-4-YL)KETONE
The alcohol (Example 5) (105 mg, 22~s ,umol) wa,s added to 5 dioxane (3 mL) and activated MnO2 (300 mg) and the black mixture wa~s stirred at reflux for 2 hr. The mixture was filtered and the clear filtrate was evaporated and the residue was chromatographed on silica gel (CHCl3-MeOH (30:1)) to yield title (35 mg).
FAB ms (M+l) 364.07.
10 Anal. Calc. for C24HI7N3O-0.35 CHCI3;
C, 72.17; H, 4.32; N, 10.37.
Found: C, 71.87; H, 4.45; N, 10.29.
1 - ~ (4-CYANOBENZYL)- 1 H-IMIDAZOL-5-Yl:,lETHYL ~ -4-PHENYL IMIDAZOLE BIS HYDROCHLORIDE SALT
N
NC~~
Step A: lH-Imidazole-4- acetic acid methyl ester hydrochloride A solution of lH-imidazole-4-acetic acid hydrochloride (4.00g, 24.6 mmol) in methanol (100 ml) wa.s saturated with ga,seous hydrogen chloride. The resulting solution was allowed to stand at room temperature for 1~ hrs. The solvent was evaporated in vacuo to afford the title compound as a white solid.
lH NMR CDC13, ~ 5(lH, ,s), 7.45(1H, s), 3.~9(2H, s) and 3.75(3H, s) ppm.
CA 0224961~ 1998-09-22 Step B: l-(Triphenylmethyl)-lH-imidazol-4-ylacetic acid methyl ester To a solution of the product from Step A (24.85g, 0.141mol) in DMF (l lSml) was added triethylamine (57.2 ml, 5 0.412mol) and triphenylmethyl bromide (55.3g, 0.171mol) and the suspension was stirred for 24 hrs. After this time, the reaction mixture was diluted with EtOAc and water. The organic phase was washed with saturated aqueous NaHCO3, dried (Na2SO4) and the solvent evaporated in vacuo. The residue was purified by chromatography (sio2~ gradient 10 elution, 0-100% EtOAc in hexanes; ) to provide the title compound as a white solid.
IH NMR CDC13, ~ 7.35(1H, s)~ 7.31(9H, m), 7.22(6H, m), 6.76(1H, s), 3.68(3H, s) and 3.60(2H, s) ppm.
~5 Step C: [1 -(4-Cyanobenzyl)- 1 H-imidazol-5-yl]acetic acid methyl ester.
To a solution of the product from Step B (8.00g, 20.9mmol) in acetonitrile (70 ml) was added 4-cyanobenzyl bromide (4.10g, 20.92 mmol) and heated at 55~C for 3 hr. After this time, the 20 reaction was cooled to room temperature and the resulting imidazolium salt was collected by filtration. The filtrate was heated at 55~C for 18hrs. The reaction mixture was cooled to room temperature and evaporated in vacuo. To the residue was added EtOAc (70 ml) and the resulting precipitate collected by filtration. The precipitated 25 imidazolium salt~s were combined, suspended in methanol (100 ml) and heated to reflux for 30 min. After thi.s time, the solvent wa~s removed in vac~o,. The resulting residue was suspended in EtOAc (75ml) and the solid isolated by filtration and washed with EtOAc.
The solid was treated with saturated aqueous NaHCO3 solution 30 (300ml) and CH2C12 (300ml) and stirred at room temperature for 2 hrs. The organic layer was separated, dried (MgSO4) and evaporated in vacuo to afford the title compound as a white solid lHNMR CDC13, ~ 7.65(1H, d, J=8Hz), 7.53(1H, s), 7.15(1H, d.
J=8Hz), 7.04(1H, s), 5.24(2H, s), 3.62(3H, s) and 3.45(2H, s) ppm.
CA 0224961~ 1998-09-22 W O 97/3~876 PCTrUS97/06257 Step D: 5-11-(4-cyanobenzyl)-lH-imidazolyllethanol.
To a stirred solution of the ester from example step C, (l.SOg, 5.8~mrnol), in methanol (20 ml) at 0~C, was added sodium 5 borohydride (l.OOg, 26.3mmol) portionwise over 5 min. The reaction was stirred at 0~C for 1 hr and then at room temperature for an additional 1 hr. The reaction was quenched by the addition of saturated NH4CI solution and the methanol evaporated in vacuo.. The residue was partitioned between EtOAc and saturated NaHC03 solution and the 10 organic extracts dried, (MgS04) and evaporated in vacuo. The residue was purified by chromatography (sio27 gradient elution, 4 to 10%
methanol in methylene chloride) to afford the title compound as a white solid.
1 H NMR CDC13 ~ 7.64(2H, d, J=8.2Hz), 7.57(1 H, s), 7.11 (2H, d, 15 J=8.2Hz), 6.97(1H, s), 5.23(2H, s), 3.79(2H, t, J=6.2Hz), 2.66(2H, t, J=6.2Hz) ppm.
Step E: 5~ (4-Cyanobenzyl~-imidazolyl)ethylmethanesulfonate.
A solution of 5-[1-(4-cyanobenzyl)-lH-imidazolyl]ethanol 20 (0.500 g, 2.20 mmol) in methylene chloride (6 ml) at 0~C was treated with Hunig's base (0.460ml, 2.64mmol) and methanesulfonyl chloride (0.204ml, 2.64mmol). After 2 hrs, the reaction was quenched by addition of saturated NaHC03 solution (SOml) and the mixture extracted with methylene chloride (50ml), dried (MgS04) and the 25 solvent evaporated in vacuo. The title compound was used without furthur purification.
lH NMR CDC13 ~ 7.69 (lH, s) 7.66(2H, d, J=8.2Hz), 7.15 (2H, d, J=8.2Hz), 7.04(1H, s), 5.24(2H, s), 4.31(2H, t, J=6.7Hz), 2.96(3H, s), and 2.88(2H, t, J=6.6Hz)ppm.
Step F~ [1 -(4-Cyanobenzyl)- 1 H-imidazol-5 -yl]ethyl ) -4-phenyl - imidazole bis hydrochloride salt.
To a suspension of sodium hydride (14.2mg, 60%
dispersion in mineral oil, 0.356mmol) in DMF (0.30 ml) at 0~C was CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 added 4-phenyl imidazole (4~.~mg, 0.339mmol), and stirred for 20 mins. A solution of the mesylate from step E (lOOmg, 0.339mmol) in DMF (0.50ml) was added to the Isolution and stirring continued at 0~C
for 1 hr and then at room temperature for 16 hrs. The reaction was 5 quenched with saturated ammonium chloride solution (O.lOml). and the the solvent evaporated in vacuo. The re~sidue was purified by chromatography (sio2~ gradient elution, 2-5% ammonium hydoxide:
acetonitrile. The resulting material was converted to the HCI salt by treating an EtOAc solution of the imidazole with gasseouls HCI and 10 evaporating the ~solvent in vacuo.
.. . . . . . .. . . .. . .. .
CA 0224961~ 1998-09-22 Anal. Calcd for C22H 19N5-2-00HCl- 1 -50H20:
C, Sg.29; H, 5.34; N, 15.45.
Found: C, 58.24; H, 5.47; N, 15.4~.
FAB HRMS exact mass calcd for C22H20N5 354.171 g71 (MH+); found 5 354.171948.
lH NMR CD30D ~ ~.93 (lH, s), 8.75(1H, s), 7.~6(1H, s), 7.76(2H, d, J=7.9Hz), 7.69(2H, d, 7.1Hz), 7.65-7.35(6H, m), 5.61(2H, s) and 4.53(2H,m)ppm.
10 In vitro inhibition of ras farne,syl transferase Assays offarnesyl-protein transferase.
Partially purified bovine FPTase and Ras peptides (Ras-CVLS, Ras-CVIM and Ras-CAIL) were prepared as described by Schaber et ah, J. Biol. Chem. 265:14701-14704 (1990), Pompliano, 15 et al., Biochemistry 31:3800 (1992) and Gibbs et al., PNAS U.S.A.
~6:6630-6634 (1989), respectively. Bovine FPTa.se was assayed in a volume of 100 ~1 containing 100 mM N-(2-hydroxy ethyl) piperazine-N'-(2-ethane sulfonic acid) (HEPl~S), pH 7.4, 5 mM MgC12, 5 mM
dithiothreitol (DTT), 100 mM [3H]-farnesyl diphosphate ([3H]-FPP; 740 20 CBq/mmol, New England Nuclear), 650 nM Ras-CVLS and 10 ~g/ml FPTase at 31 ~C for 60 min. 3~eactions were initiated with FPTase and stopped with 1 ml of 1.0 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 ,~-plate counter. The assay 25 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 radioactivity in the 30 presence of the test compound when compared to the amount of incorporation in the absence of the test compound.
- Human FPTa~se was prepared as described by Omer et al., Biochemistry 32:5167-5176 (1993). Human FPTase activity was assayed as described above with the exception that 0.1 % (w/v) CA 0224961~ 1998-09-22 W O 97/36876 PCTnUS97/06257 polyethylene glycol 20,000, 10 ~lM ZnC12 and 100 nM Ras-CVIM were added to the reaction mixture. Reactions were performed for 30 min., stopped with 100 ~1 of 30% (v/v) trichloroacetic acid (TCA) in ethanol and processed as described above for the bovine enzyme.
S The compounds of the instant invention described in the above Examples 1-7 were tested for inhibitory activity against human FPTase by the assay described above and were found to have IC50 of 50 ~lM.
In vivo ras farnesylation assay The cell line u~ed in this assay is a v-ras line derived from either Ratl or NIH3T3 cells, which expressed viral Ha-ras p21.
The assay is performed essentially as described in DeClue, J.E. et ah, Cancer Research 51 :712-717, (1991). Cells 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 labelled in 3 ml methionine-free DMEM supple-meted with 10% regular DMEM, 2~o fetal bovine serum and 400 mCi[35S]methionine (1000 Ci/mmol). After an additional 20 hours, the cells are lysed 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 count.s are bought to 1 ml with IP buffer (lysis buffer lacking DTT) and immunoprecipitated with the ra~s-specific monoclonal antibody Y13-259 (Furth, M.E. et ah, J. Virol. 43:294-304, (19X2)). 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/l % 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 autoradiographed. The intensities of the bands corresponding to CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/06257 farne~ylated and nonfarnesylated ras proteins are compared to determine the percent inhibition of farnesyl transfer to protein.
In viv(~ ~rowth inhibition assay To determine the biological consequences of FPTase inhibition, the effect of the compounds of the instant invention on the anchorage-independent growth of Ratl cells transformed with either a v-ras, v-7~af, or v-mos oncogene is tested. Cells transformed 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 are seeded at a density 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 appropriate concentration of the instant compound (dissolved 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.
Another preferred group of compounds includes the compounds of formula I wherein A3 represents aryl or heteroaryl.
Preferably R6 represents CN.
Preferably, R7represents hydrogen, unsubstituted or substituted Cl-C6 alkyl.
Preferably, RX represent.s H or Cl ~ alkyl, and R9 i.s C
- alkyl.
Preferably, Al and A2 are independently selected from:
CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 a bond, -C(O)NRX-, -NR~C(O)-, -O-, -N(RX)-, -S(O)2N(RX)- and-N(Rg)S(0)2--- Preferably, V is selected from hydrogen, heterocyclyl and aryl. More preferably V is phenyl.
Preferably, W is heterocyclyl selected from imidazolinyl, imidazolyl, oxazolyl, pyrazolyl, pyyrolidinyl, thiazolyl and pyridyl.
More preferably, W i.s selected from imidazolyl and pyridyl.
Preferably X represents aryl. In particular, X can represent phenyl.
Preferably, m is 0 or 2.
Preferably n and p are 0, 1, 2 or 3.
A subset of compounds of the invention is represented by forrnula Ia:
(R )r V - A1 (C R 1 a2)nA2(C R 1 b2)~3 N ~R3 (CR22)p--A3- (CR22)p X
\ 4 15 wherein:
R3, R4, A3, R~, R9, X, m, n, p and r are as originally defined;
each R 1 a and R2 is independently selected from hydrogen and Cl -C6 alkyl;
each Rlb is independently selected from: hydrogen, aryl, heterocyclyl, C3 1() cycloalkyl, C2-~ alkenyl, R~O-, -N(R8)2 and Cl-C6 alkyl unsubstituted or substituted by aryl, heterocyclyl, cycloalkyl, alkenyl, RgO- and -N(RX)2;
CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 R6 is independently selected from: hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-c6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl, R80-, R~C(O)NR~-, CN, NO2, (R~s)2N-c(NR~ R~sC(O)-, R~OC(O)-, -N(Rg)2, or R9OC(o)NR8-, and Cl-C6 alkyl substituted by Cl-C6 5 perfluoroalkyl, R80-, R8C(O)NR8-, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2 and R9OC(o)NR8-;
R7 repre.sents H or Cl-6 alkyl;
A 1 and A2 are independently selected from: a bond, -CH=CH-, -C_C-, -C(O)-, -C(O)NR2~-, O, -N(R8)- and S(O)m;
and V is selected from: hydrogen; aryl; heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, ~uinolinyl, isoquinolinyl and thienyl; Cl-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if Al is a bond and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R6 and to Al is through a substitutable ring carbon.
A second subset of compounds of the present invention is represented by formula Ib:
(R6) /~7~ / R3 V-A1(CR1a2)nA2(CR1b2)n-\W~-(CR22)p-R3C=CR8~CR22),~X
Ib wherein:
Rla Rlb, R2, Al, A2, R3, R4, R6, RX, R9, X, m, n, p and r are as originally defined;
R7 is selected from: hydrogen and Cl-C6 alkyl;
CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/06257 V is selected from: hydrogen, heterocyclyl ~elected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl and thienyl; aryl; Cl-c2o alkyl wherein from 0 to 4 carbon atom.s are replaced with a heteroatom selected from O, S, and N, S and C2-C20 alkenyl, provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if A 1 is a bond, n i~s 0 and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R~ and to Al is 10 through a substitutable ring carbon;
and W represents heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl and isoquinolinyl.
A third subset of compounds of the present invention is repre~ented by formula Ic:
(R6) (17~ ~R3 V-A1(CR1a2)nA2(CR1b2)n\W~~(CR22)pC_C--(CR2z~X
Ic R4 wherein:
Rl~ Rlb R2, Al, A2, R3, R4, R6, RX, R9, X, m, n, p and r are as originally defined;
R7 is selected from: hydrogen and Cl-C6 alkyl;
V is selected from: hydrogen, heterocyclyl ,selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, aryl, Cl-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, CA 0224961~ 1998-09-22 provided that V is not hydrogen if Al is S(O)m and V is not hydrogen if Al i~sabond,n isOandA2isS(O)m;
provided that when V is heterocycle, attachment of V to R~ and to Al is 5 through a substitutable ring carbon;
and W represents heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl and i.soquinolinyl.
A fourth embodiment of the invention is described in accordance with formula Id:
H
)~N
,=~N~\J ~3 ~¦~ (CR 2)p-A - (CR 2)p X~
n Id wherein:
each R2 is independently selected from hydrogen and Cl-C6 alkyl;
R3, R4, A3, RX, R9, X, m and p are as originally defined;
and R6 is selected from the group consisting of: hydrogen, Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl, F, Cl, R~O-, R8C(O)NR8-, CN, NO2, (R8)2N-C(NR8)-, R8C(O)-, 25 R8OC(O)-, -N(R8)2, or R9OC(o)NR~- and Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl, R80-, R8C(O)NR~-, (R8)2N-C(NR8)-, R8C(O), R8OC(O)-, -N(R~)2 or R9OC(o)NRX-.
W O 97/36876 PCT~US97/06257 A fifth sub.set of compounds of the invention is represented by formula le:
H
)~N
(C R22)p A3 - (C R22)--X
NC
le wherein:
X and A3 are as originally defined;
each R2 is independently selected from: hydrogen and Cl-C6 alkyl;
R3 and R4 are independently ,selected from H, F, Cl, Br, N(R8)2, CF3, N02, (R8)o-, (R9)S(o)m-, (R8)C(O)NH-, H2N-C(NH)-, (R8)C(O)-, (R8)0C(O)-, N3, CN, (R9)OC(O)NR8-, Cl-C20 alkyl, substituted or unsub.stituted aryl, and substituted or unsubstituted heterocyclyl;
and R~, R9, m and p are a.s originally defined.
Specific examples of compound.s of the invention are:
NC~ C--C~
<'~
N
PCTrUS97/06257 NC ~ CH--CH~
N
NC ~ C--C
N
~\N ~ C H = C H ~ ~
NHo Cl NC ~N~C _C
N
NC~3~N
NC ~ N~
N
~>~N~? CH2-C--C
N
NC ~C--C
~\N~
S N
~_N CN
N
OH
N
' ~
N c'(3J ~3 CA 0224961~ 1998-09-22 and the pharmaceutically acceptable salts and isomers thereof.
The pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds 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, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.
The pharmaceutically acceptable salts of the compounds of this invention can be synthesized from the compounds of this invention which contain a basic moiety by conventional chemical methods. Generally, the salts are prepared either by ion exchange chromatography or by reacting the free base with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid in a suitable ,solvent or variou,s combinations of solvents.
Reactions used to generate the compounds of this invention are prepared by employing reactions as shown in the Schemes, 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. Substituents R and R CH2-, as shown in the Schemes, represent the substituents R8, R9 and others, depending on the compound of the instant invention that is being synthesized. The variable p' represents p-l.
These reactions may be employed in a linear sequence to provide the compounds of the invention or they may be u,sed to synthesize fragments which are subsequently joined by the alkylation reactions described in the Schemes.
Svnopsis of Schemes CA 0224961~ 1998-09-22 The requisite intermediates are in ~some cases commercially available, or can be prepared according to literature procedures.
Schemes 1-2 illustrate the synthesi.s of one of the preferred embod~-ment.s of the instant invention, wherein the variable W is present as an imidazolyl moiety that is sub,stituted with a suitably sub,stituted benzyl group. Substituted protected imidazoles can be prepared by method.s such a,s those de.scribed by F. Schneider, Z. Physiol. Chem., 3:206-210 (1961) and C.P. Stewart, Biochem. Journal, 17:130-133(1923).
Benzylation and deprotection of the imidazole alkanol provide.s intermediate III which can be oxidized to the corresponding aldehyde IV. Also, while X is shown as a phenyl ring, other aryl and heteroaryl groups can be substituted therein without departing from the invention.
The a~dehyde whose synthesis is illustrated in Scheme 1 may be reacted with ~ suitably substituted aralkyne, to provide the intermediate compound V. Compound V can be selectively hydrogenated across the unsaturated bond under standard conditions, such as those illustrated, to provide Compound VI.
Schemes 3- 10 illustrate syntheses of suitably substituted aldehydes u,seful in the syntheses of the instant compounds wherein the variable W is a pyridyl moiety. Similar synthetic strategies for preparing alkanols that incorporate other heterocyclic moieties for variable W can be discerned from the teachings herein.
Generally the aldehyde i,s reacted with an appropriately substituted aralkyne using n-BuLi, after which the triple bond can be reduced. As shown in Schemes 2, 4, 6 and ~s reduction of the alkyne triple bond using Pd/BaSO4 Iproduces the Z-olefin isomer almost exclusively. By substituting sodium bi,~(2-methoxyethoxy)aluminum hydride (RED-AL) in toluene, one can readily obtain the E-allylic alcohol from propargylic alkynes used in the present invention.
In the preparation methods described herein, reactive groups may remain blocked until the final product is prepared, - essentially in protected form, after which a final deprotection step is conducted. These blocking groups are readily removable i.e., they can be removed, if desired, by procedures which will not cause cleavage or CA 0224961~ 1998-09-22 other disruption of the remaining portions of the molecule. Such procedures include chemical and enzymatic hydrolysis, treatment with chemical reducing or oxidizing agents under mild conditions, treatment with fluoride ion, treatment with a transition metal catalyst and a 5 nucleophile and catalytic hydrogenation.
Examples of suitable hydroxyl protecting groups are:
t-butylmethoxyphenylsilyl, t-butoxydiphenylsilyl, trimethylsilyl, triethylsilyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, benzyloxycarbonyl, t-butyloxycarbonyl, 2,2,2-trichloroethyloxy-10 carbonyl and allyloxycarbonyl. Preferred hydroxyl protecting groupsare trimethylsilyl and triethylsilyl.
Examples of suitable carboxyl protecting groups are:
benzhydryl, o-nitrobenzyl, p-nitrobenzyl, 2-naphthylmethyl, allyl, 2-chloroallyl, benzyl, 2,2,2-trichloroethyl, trimethylsilyl, 15 t-butyldimethylsilyl, t-butyldiphenylsilyl, 2-(trimethylsilyl)ethyl, phenacyl, p-methoxybenzyl, acetonyl, p-methoxyphenyl, 4-pyridylmethyl and t-butyl. A preferred-carboxyl protecting group is p-nitrobenzyl.
Many other suitable hydroxyl and carboxyl protecting 20 groups are known in the art. See, e.g., T.W. Greene, Protective Groups in Or~anic Synthesis, John Wiley & Sons, Inc., 1981 (Chapters 2 and 5).
W O 97/36876 PCT~US97/06257 ~(cR22)Pl-cH2oH Prot1X Et3N Prot 'N~/~>
H DMF
lla rBr (C R22)p -C H20Ac R 6~EtOAc AC20, Py ,=1=~
Prot1~N~ N 2. N-deprotect CR2 ) -CH OAC N~(CR 2)p-CH20H
N~ LiOH
R6/~ THF, H20 ~
N,~(CR 2)p,-CHO HO r/R3 ~N ~/ (CR22)p.-CH-C_C~_~
SO3Py, Et3N ~ n-BuLi, THF ~ > R4 DMSO~ H
V
W O 97/36876 PCT~US97/06257 )p.-CH-C_C{;~
~N R4 R6 H2, Pd/C
V
MeOH, EtOAc H2, Pd/BaSO4 MeOH, EtOAc HO R3 HO 3,~cR22)p-cH
~$CR22)p~-cH-CH=c~_;~ N
~N R4 (predominantly R6/~
~, Z isomer) Vlb R6/~
Vla CH3 1) HN~2~Br2 CO2CH3 ~ 2) KMnO
H2N N 3) MeOH,H+ Br N
r ~\MgCI R6 ~ ~,C02CH3 ZnCI2,NiCI2(Ph3P)2 N
NaBH4 (excess) ~,CH20H
SO3 Py Et3N ~CHO
n-BuLi,THF R6 H--C--C{ ~ ~)~CH
W O 97/36876 PCTrUS97/06257 H2, Pd/BaSO4 MeOH, EtOAc H2, Pd/C
~" MeOH, EtOAc R6 ~CH-CH=CH~
(predominantlyZ isomer) ~ ~CH~ R4 W O 97136876 PCTrUS97/06257 1. EtOC(=O)CI
~,CO2CH3 ~Z\ 9C CN ~C02CH3 N 3. S, xylene, heat N
NaBH4 ~ SO3Py, Et3N ~
(excess) ~"CH20H DMSO ~CHO
Br CO2CH3[~\ MgCI [~j ZnCI2, Nicl2(ph3p)2 ~CO2CH3 NaBH4 ¦ SO3Py, Et3N
~CH20H ~ ~CHO
(excess) N DMSO NJJ
N-BuLi, THF ll f, OH
H = ~R~
OH
~,CH
N
H2, Pd/BaSO4 H2, Pd/C
MeOH, EtOAc MeOH, EtOAc ¢~3,CH-CH=CH~ 3,OH
(predominantly Z isomer) Br~ 1. LDA, CO2 Br~
N 2. MeOH, H+ N
r ZnC12, NiC12(Ph3P)2 N
NaBH4 (excess) ~ ( ~20H S03Py, Et3N
DMSO
R~
N
H2, Pd/BaSO4 H2, Pd/C
MeOH, EtOAc MeOH, EtOAc ~' N N
(predominantly Z isomer) W O 97/36876 PCTrUS97/06257 1. LDA, CO2 [~Br 2. (CH3)3sicHN2 R6 ~Br R6 ~
~, co2CH3 Zn, NiC12(Ph3P)2 N~
R6 ~
excess NaBH4 ~ SO3 Py, Et3N
N ~CH20H DMSO
~, R6 ~ R6 ~ OH
~CHO n-BuLi, THF N~ h R3 -W O 97t36876 PCTrUS97106257 R6 ~q OH
~- = 7 H2, Pd/C R4 MeOH, EtOAc R6 ~
~/
BocN H ~
BocNH CHO
n-BuLi, THF
BocN H
/=~ R CF3CO2H
BocNH~ ~ R4 CH2CI2 OH
R3 Boc20 NH2/~ R4 CH2CI2 OH
BocNH (R )r~ CHO
NH2~ R4 NaBH(OAc)3 OH Et3N, CICH2CH2CI
BocN H
(R )r~--CH2 ~ ~:~R3 W O 97/36876 PCT~US97/06257 BocN H
~ R CF3CO2H
(R )~ CH 'NH~ R4 NaHCO3 NH~ ~R
(R )r~--CH2 ~ R4 AgCN
(R )r~ CH/ \~ 4 H ~ R3 Me><O~ ~
Me N~\cHo R4 Boc n-BuLi, TH F
>< ~ R3 t-Bu(Me)2Si-CI
, ~R4 imidazole, DMF
Boc OH
Me>< ~ R3 TsOH, H20 Boc R benzene O-SiMe2(t-Bu) HO
~ R CICOCOCI
BocNH~ R4 DMSO CH2C12 O-SiMe2(t-Bu) (c2H5)3N
O H
R3 1. R'MgX
BocNH--~R4 2. TFA
O-SiMe2(t-Bu) CH2CI2 HO R' ~ ~/ R
H2N~ R4 OH
HO
~ R CF3CO2H
BocNH~ R4 CH2C12 O-SiMe2(t-Bu) HO
~ R3 R'CHO
H2N~ R4 NaBH(OAc)3 HO
R'CH2~N A,~ R4 OH
HO 3 N=~ ~= N
=~ R ~ ' S ~
BocNH~ ~ R4 ~2 ,, O-SiMe2(t-Bu) NaH, DMF 0~C
Boc--N1 ~=~ R3 R'SH
4 (C2H5)3N ~, R CH3oH
O-SiMe2(t-Bu) R'S
~ R CF3CO2H
BocNH '\~ R4 CH2C12 O-SiMe2(t-Bu) R'S ~ R3 H2N~ R4 OH
W O 97/36876 PCTrUS97/06257 HO,~1) Boc20, K2C~3 HO~
~/ THF-H20 2) CH2N2, EtOAc J~
H2NC02H BocNH CO2CH3 HO,,~
LiAlH4 ,b~l R'CH2X
THF l Cs2CO3 0-20~C BocNH CH2OH DMF
R'C H20 R'C H20 )~ pyridine SO
DMSO
BocNH CH20H (c2H5)3N BocNH~CHO
WO 97136876 PCTnUS97/062~7 SCHEME 16 (cont.) R'CH ;~
BocNH CHO n-BuLi, THF
R'CH20~
BocNH J~ _ lc/R3 HCI / \ BF3-0Et2, EtSH
ETOAc / \ CH2CI2 R'C H20~3 HO~
H2NJ~ ~R H2NJ~
The instant compound.s are useful in the treatment of cancer. Cancers which may be treated with the compounds of this 5 invention include, but are not limited to, colorectal carcinoma, exocrine pancreatic carcinoma, myeloid leukemia~ and neurological tumor.s. Such tumors may arise by mutations in the ra~ genes CA 0224961~ 1998-09-22 W O 97136876 PCTrUS97106257 themselves, mutations in the proteins that can regulate Ras activity (i.e., neurofibromin (NF-l), neu, scr, abl, lck, fyn) or by other mechani~sms.
The compounds of the instant invention inhibit farnesyl-protein transferase and farnesylation of the oncogene protein Ras.
5 The instant compounds may also inhibit tumor angiogenesis, thereby affecting the growth of tumors (J. Rak et al. Cancer Researc~h, 55:4575-4580 (1995)). Such anti-angiogenic 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 diseases where Ras proteins are aberrantly activated a,s a result of oncogenic mutation in other genes (i.e., the Ras gene itself i.s 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 component of NF-l is a benign proliferative disorder.
The instant compounds may also be useful in the treatment of 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 the prevention of restenosis after percutaneous transluminal coronary angioplasty by inhibiting neointimal formation (C. Indolfi et al. Natu~ e medicine, I :541 -545(1995).
The instant compounds may also be useful in the treatment and prevention of polycystic kidney disease (D.L. Schaffner et al.
Ame~ican Journal of Pathology, 142:1051-1060 (1993) and B. Cowley, Jr. et al.FASFB Journal, 2:A3160 (19~s~)).
The instant compounds may also be useful for the treatment of fungal infections.
The compounds of this invention may be administered to m~mm~ls, preferably humans, either alone or, preferably, in combina-- tion with pharmaceutically acceptable carriers or diluents, in the form of a pharmaceutical composition, which is comprised of a compound of formula I in combination with a pharmaceutically acceptable carrier.
CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/06257 The compounds can be administered orally, topically, rectally, vaginally transderrnally or parenterally, including the intravenous, intramuscular, intraperitoneal and ,subcutaneous routes of administration.
For oral use, the compound is administered, for example, 5 in the form of tablets or capsules, or as a solution or .suspen,sion. In the case of tablets for oral use, carriers which are commonly used include lactose and corn ,starch; lubricating agents, such as magnesium stearate, are commonly added. For oral administration in capsule form, diluents also include lactose and dried corn starch. When a4ueous suspensions 10 are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If de,sired, certain sweetening and/or flavoring agents may be added. For intramuscular, intraperi-toneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared, the pH of the solution is suitably 15 adjusted and the product is buffered. For intravenous use, the total concentration is controlled to render the preparation substantially isotonic.
As used herein, the term "composition" i.s intended to encompass a product comprising the specified ingredients in the specific 20 amounts, as well as any product which results, directly or indirectly, from combination of the specific ingredients in the specified amount.s.
The compound,s of the instant invention may also be co-~lmini~tered in therapeutic composition~s that also contain other well known therapeutic agents that are selected for their particular 25 usefulness against the condition that is being treated. For example, the instant compound.s may be useful in combination with known anti-cancer and cytotoxic agents. Similarly, the instant compounds may be u.seful in combination with agents that are effective in the treatment and prevention of NF-I, restinosis, polycystic kidney disease, infection.s of 30 hepatitis delta and related viruses and fungal infections.
If formulated a~s a fixed dose, such combination products - employ a compound of this invention substantially within the dosage range described below and other pharmaceutically active agent(s~
typically within the acceptable dosage range. Compounds of the CA 0224961~ 1998-09-22 W O 97t36876 PCTrUS97/06257 -3~-instant invention may alternatively be used sequentially with known pharmaceutically acceptable agent(s) when a combination formulation i.s inappropriate.
The daily dosage will normally be determined by the 5 prescribing physician, who may vary the dosage according to the age, weight, and response of the individual patient, a.s well as the severity of the patient's condition.
In one exemplary application, a suitable amount of compound is administered to a m~mm~l undergoing treatment for 10 cancer. Adrnini.stration occurs in an amount between about 0.1 mg/kg of body weight to about 60 mg/kg of body weight per day, preferably of between 0.5 mg/l~g of body weight to about 40 mgtkg of body weight per day.
The compounds of the instant invention are also useful 15 as a component in an assay to rapidly determine the presence and quantity of farnesyl-protein transferase (FPTase) in a composition.
Thus the composition to be tested may be divided and the two portions contacted with mixtures which comprise a known substrate of FPTase (for example a tetrapeptide having a cysteine at the amine 20 terminus) 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 farnesylate the substrate, the chemical content of the assay mixture.s may be determined by well known immuno-25 logical, radiochemical orchromatographic techniques. Becausethe 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 compound of the instant invention relative to the presence of the unchanged sub~strate in the a,ssay 30 containing the instant compound i.s 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 u~seful in identifying tissue samples which contain farnesyl-protein transferase and quanti-CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/062~7 tating the enzyme. Thus, potent inhibitor compounds of the instant invention may be u~sed in an active site titration assay to determine the quantity of enzyme in the sample. A serie~s of .samples composed of aliquots of a tissue extract containing an unknown amount of farne,syl-5 protein transferase, an excess amount of a known substrate of FPTa~se (for example a 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 10 that has a Ki Isubstantially 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.
(+)-1 -(4-CYANOBENZYL)-S-1 ( I -HYDROXY-3-PHENYL)-2-PROPYNYLlIMIDAZOLE HYDROCHLORIDE
20 Step A: Preparation of l-triphenylmethyl-4-(hydroxymethyl)imidazole To a solution of 4-(hydroxymethyl)imidazole hydrochloride (35.0 g, 260 mmol) in 250 mL of dry DMF at room temperature was added triethylamine (90.6 mL, 650 mmol). A white 25 solid precipitated from the solution. Chlorotriphenylmethane (76.1 g, 273 mmol) in 500 mL of DMF was added dropwise. The reaction mixture was .stirred for 20 hour.s, poured over ice, filtered, and washed with ice water. The resulting product wa,s slurried with cold dioxane, filtered, and dried in vacuo to provide the titled product as a white solid 30 which was sufficiently pure for use in the next ,step.
Step B: Preparation of l-triphenylmethyl-4-(acetoxymethyl)imidazole CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 Alcohol from Step A (260 mmol, prepared above) was suspended in 500 mL of pyridine. Acetic anhydride (74 mL, 7~0 mmol) wa.s added dropwi.se, and the reaction was stirred for 4~ hours during which it became homogeneous. The solution was poured into 2 S L of EtOAc, washed with water (3 x 1 L), 5% aq. HCI soln. (2 x 1 L), sat. aq. NaHCO3, and brine, then dried (Na2SO4), filtered, and concentrated in vacuo to provide the crude product. The acetate was isolated a~s a white powder (85.X g, 86% yield for two ~teps) which wa~s sufficiently pure for use in the next reaction.
Step C: Preparation of 1-(4-cyanobenzyl)-S-(acetoxymethyl)imidazole hydrobromide A solution of the product from Step B (85.8 g, 225 mmol) and ~-bromo-p-tolunitrile (50.1 g, 232 mmol) in 500 mL of EtOAc was lS stirred at 60 ~C for 20 hours, during which a pale yellow precipitate formed. The reaction wa.s cooled to room temperature and filtered to provide the solid imidazolium bromide salt. The filtrate was concen-trated in vacuo to a volume 200 mL, reheated at 60~C for two hours, cooled to room temperature, and filtered again. The filtrate was 20 concentrated in vacuo to a volume 100 mL, reheated at 60~C for another two hours, cooled to room temperature, and concentrated in vacuo to provide a pale yellow solid. All of the solid material was combined, dissolved in 500 mL of methanol, and warmed to 60~C.
After two hours, the solution was reconcentrated in V~lCUo to provide 25 a white solid which was triturated with hexane to remove soluble materials. Removal of residual solvents in vacuo provided the titled product hydrobromide a~s a white solid (50.4 g, 67% yield, ~9% purity by HPLC) which was used in the next step without further purification.
30 Step D: Preparation of 1-(4-cyanobenzyl)-S-(hydroxymethyl)imidazole - To a solution of the acetate from Step C (50.4 g, 150 mmol) in l .S L of 3: I THF/water at 0 ~C was added lithium hydroxide monohydrate (18.9 g, 450 mmol). After one hour, the reaction was CA 0224961~ 1998-09-22 concentrated in vacuo, diluted with EtOAc (3 L), and washed with water, .sat. a~. NaHCO~ and brine. The solution was then dried (Na2SO4), filtered, and concentrated in vacuo to provide the crude product (26.2 g, ~2% yield) as a pale yellow fluffy solid which was ~S sufficiently pure for use in the next ~itep without further purification.
Step E: Preparation of 1-(4-cyanobenzyl)-~S-imidazolecarboxaldehyde To a solution of the alcohol from Step D (21.5 g, 101 mmol) in 500 mL of DMSO at room temperature was added triethylamine (56 mL, 402 mmol), then SO3-pyridine complex (40.5 g, 254 mmol). After 45 minutes, the reaction was poured into 2.5 L
of EtOAc, washed with water (4 x 1 L) and brine, dried (Na2SO4), filtered, and concentrated in vacuo to provide the aldehyde (1~.7 g, 1~ ~8% yield) as a white powder which was sufficiently pure for use in the next step without further purification.
Step F: Preparation of (+)-1-(4-cyanobenzyl)-5-[(1-hydroxy-3-phenyl)-2-propynyllimidazole hydrochloride To a solution of phenylacetylene (0.172 mL, 1.56 mmol) in 5 mL of THF at 0 ~C was added n-butyllithium (0.530 mL, 2.5 M in hexanes, 1.32 mmol). After 15 minutes, the aldehyde from Step E (254 mg, 1.20 mmol) was added, and the reaction was stirred for 30 minutes.
The reaction was quenched with sat. aq. NaHCO3, poured into EtOAc, washed with sat. aq. NaHCO3 and brine, dried (Na2SO4), filtered, and concentrated in vacuo. The resulting product was purified by silica gel chromatography (35-50% acetone/CH2CI2) to provide 1 ~7 mg of the desired alcohol. A portion of this was taken up in CH2CI2 and treated with excess 1 M HCI/ether solution, and concentrated in vacuo. The titled product hydrochloride wa.s isolated as a white ~olid.
- FAB mas.~i spectrum m/e 314 (M+l).
Analysis calculated for C2~HI~N3O ~ 1.0 HCI - 0.90 H2O:
C, 65.63; H, 4.90; N, 11.4~;
W O 97/36876 PCTrUS97/06257 Found: C, 65.81; H, 4.98; N, 1 1.17.
(+)-1 -(4-CYANOBENZYL)-5-~( 1 -HYDROXY-4-PHENYL)-3-BUTYNYLlIMIDAZOLE HYDROCHLORIDE
To a solution of t-butyllithium in 1.5 mL of THF (0.78 mL
of 1.7 M in pentane, 1.32 mmol) at -78~C was added tetramethylethyl-enediamine (0.199 mL, 1.32 mmol) and 1-phenyl-1-propyne (0.150 mL, 1.20 mmol). The solution was warmed to 0 ~C for one hour, then cooled to -78 ~C. The aldehyde from Step E of Example 1 (225 mg, 1.07 mmol) on 1.0 mL THF was added, and the reaction allowed to warm to 0 ~C. After 30 minutes, the reaction was quenched with sat.
aq. NaHCO3, poured into EtOAc, washed with sat. aq. NaHCO3 and brine, dried (Na2SO4), filtered, and concentrated in vacuo. The product was purified by silica gel chromatography (3-4% MeOH/CH2CI2), then taken up in CH2CI2 and treated with excess 1 M HCI/ether solution, and concentrated in vacuo to provide the titled product hydrochloride (48 mg) as a pale yellow foam.
FAB mass spectrum m/e 32~ (M+l).
Analysis calculated for C2lH17N3O ~ 1.10 HCl ~ 0.10 Et2O:
C, 68.56; H, 5.14; N, 11.21;
Found: C, 68.30; H, 5.04; N, 11.06.
(+)-3-(4-CYANOBENZYL)-4-l ( 1 -HYDROXY-3-PHENYL)-2-PROPYNYLlPYRIDINE HYDROCHLORIDE
- Step A: Preparation of 3-(4-cyanobenzyl)pyridin-4-carboxylic acid methyl ester CA 0224961~ 1998-09-22 W 097/36876 PCTrUS97/06257 A solution of 4-cyanobenzyl bromide (625 mg, 3.27 mmol) in dry THF (4mL) was added slowly over~3 min. to a suspension of activated Zn (dust; 250 mg) in dry THF (2 mL) at O~ under an argon atmosphere. The ice-bath was removed and the slurry was stirred at S room temperature for a further 30 min. Then 3-bromopyridin-4-carboxylic acid methyl ester (~40 mg. 2.5 mmol) followed by dichlorobis(triphenylphosphine)nickel (Il) (50 mg). The resultant reddish-brown mixture was stirred for 3h at ~40-45~C. The mixture was cooled and distributed between EtOAc (l00 ml) and 5% aqueou~
citric acid (50 mL). The organic layer was washed with H2O (2X50 mL), dried with Na2SO4. After evaporation of the solvent the residue was purified on silica gel, eluting with 35% EtOAc in hexane to give 420 mg as a clear gum. FAB ms (M+l) 253.
Step B: Preparation of 3-(4-cyanobenzyl)-4-(hydroxymethyl)pyridine The title compound was obtained by sodium borohydride (300 mg) reduction of the ester from Step A (415 mg) in methanol (5 mL) at room temperature. After stirring for 4 h the solution was evaporated and the product was purified on silica gel, eluting with 2%
methanol in chloroform to give the title compound. FAB ms (M+l ) 225.
Step C: Preparation of 3-(4-cyanobenzyl)-4-pyridinal The title compound was obtained by activated manganese dioxide (1.0g) oxidation of the alcohol from Step B (240 mg, 1.07 mmol) in dioxane (10 mL) at reflux for 30 min. Filtration and evaporation of the solvent provided title compound, mp ~0-~s3OC.
Step D: Preparation of (+)-3-(4-cyanobenzyl)-4-[(1-hydroxy-3-phenyl)-2-propynyllpyridine hydrochloride The titled compound is prepared from the pyridinal from Step C using the procedured in Step F of Example 1. The product is purified by silica gel chromatography, then taken up in CH2CI2 and CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 treated with excess 1 M HCl/ether solution, and concentrated i~ cuo to provide the titled product hydrochloride.
1 -(4-BIPHENYLMETHYL)-5-(4-CYANOBENZYL)IMIDAZOLE
HYDROCHLORIDE SALT
Step A: 1 -Trityl-4-(4-Cyanobenzyl)-imidazole.
To a suspension of activated zinc dust (3.57g, 54.98mmol) in THF (50ml)was added dibromoethane (0.315m1, 3.60mmol) and the reaction stirred under argon at 20~C. The suspension was cooled to 0~C
and (x bromo-p-toluinitrile (9.33g, 47.6mmol) in THF (lOOml) was added dropwise over a period of 10 min. The reaction was then allowed to stir at 20~C for 6hr and bi,s(triphenylphosphine)Nickel II chloride (2.4g, 3.64mmol) and 5 -iodotrityl imidazole (15.95g, 36.6mmol) was added in one portion.The resulting mixture was stirred 16hr at 20~C
and then quenched by addition of saturated NH4CI solution (lOOml) and the mixture stirred for 2 hours. Saturated NaHCO3 solution was added to give a pH of P~ and the solution wa,s extracted with EtOAc (2x250ml), dried MgSO4 and the solvent evaporated in vacuo. The residue was chromatographed (sio2~ 0-20% EtOAc/CH2Cl2 to afford the title compound as a white solid.
1H NMR ~ CDC13 (7.54 (2H, d, J=7.9Hz), 7.38(1H, s), 7.36-7.29 (1 lH, m), 7.15-7.09(6H, m), 6.58(1H, s), and 3.93(2H, s)ppm.
Step B: 1 -(4-Biphenylmethyl)-5-(4-cyanobenzyl)imidazole hydochloride salt To 1-Trityl-4-(4-Cyanobenzyl)-imidazole (60~.~ mg, 1.43 mmol) in acetonitrile (2 ml) was added 4-chloromethyl biphenyl (290mg, 1.43 mmol) and the mixture heated at 55~C for 16 hours. The - residue was dissolved in methanol (30 ml) and heated at reflux for 20 mins, cooled and evaporated to dryness. The residue was partitioned between saturated NaHCO3 solution and CH2C12. The organic layer was dried (MgSO4) and the solvent evaporated in vacuo. The re~idue wa~ chromatographed (SiO2, 5% methanol in CH2C12) to afford the imidazole which was converted to the HCI salt by treatment with one equivalent of HCI in a4ueous acetonitrile. Evaporation of Isolvent in 5 vacuo afforded the title compound as a white powder.
Anal. Calcd for C24HlgN3- 1.00 HCI:
C, 74.70; H, 5.22; N, 10.~9.
Found: C, 74.70; H, 5.31; N, 10.77.
FAB MS 350 (MH+) 10 lH NMR CD30D ~ 9.03(1H, s), 7.65-7.50(5H, m), 7.44(2H, t, J=7.5Hz), 7.39(1H, s), 7.35(1H, t, J=7.3Hz), 7.26(2H, d, J=~.lHz), 7.20(2H, d, J=X.lHz), 5.42(2H, s), and 4.17(2H, ,s) ppm.
I l -(4-CYANOBENZYL)IMIDAZOL-5-YLl(~ 1.1 '-BIPHENYLl-4-YL)METHANOL
A Grignard reagent, freshly prepared from 4-bromo[l,l'-20 biphenyl] (116 mg, 500 ,umol) and magnesium turnings (1~ mg, 730 ,umol) in dry THF (500 ,ul) was added to a dry Argon-purged 3mL flask containing the aldehyde (105 mg, 500 }lmol) in dry THF (200 ~L) with vigorous stirring at room temperature. After I hour the reaction was quenched with sat. NH4CI (5 mL) and distributed between EtOAc (50 25 mL) and H2O (50 mL). The organic phase was evaporated and the residue was chromatographed on silica gel (CHCI3-MeOH (20:1)) to yield title (117 mg).
FAB ms (M+l) 366.25.
Anal. Calc. for C24H IgN3O-0.10 CHCI3-0.10 CH3OH;
C, 76.37: H, 5.16: N, 11.04.
Found: C, 76.13; H, 5:10; N, 10.76.
W O 97/36876 PCTrUS97/06257 11-(4-CYANOBENZYL)IMIDAZOL-5-YLl(l l.l'-BIPHENYLl-4-YL)KETONE
The alcohol (Example 5) (105 mg, 22~s ,umol) wa,s added to 5 dioxane (3 mL) and activated MnO2 (300 mg) and the black mixture wa~s stirred at reflux for 2 hr. The mixture was filtered and the clear filtrate was evaporated and the residue was chromatographed on silica gel (CHCl3-MeOH (30:1)) to yield title (35 mg).
FAB ms (M+l) 364.07.
10 Anal. Calc. for C24HI7N3O-0.35 CHCI3;
C, 72.17; H, 4.32; N, 10.37.
Found: C, 71.87; H, 4.45; N, 10.29.
1 - ~ (4-CYANOBENZYL)- 1 H-IMIDAZOL-5-Yl:,lETHYL ~ -4-PHENYL IMIDAZOLE BIS HYDROCHLORIDE SALT
N
NC~~
Step A: lH-Imidazole-4- acetic acid methyl ester hydrochloride A solution of lH-imidazole-4-acetic acid hydrochloride (4.00g, 24.6 mmol) in methanol (100 ml) wa.s saturated with ga,seous hydrogen chloride. The resulting solution was allowed to stand at room temperature for 1~ hrs. The solvent was evaporated in vacuo to afford the title compound as a white solid.
lH NMR CDC13, ~ 5(lH, ,s), 7.45(1H, s), 3.~9(2H, s) and 3.75(3H, s) ppm.
CA 0224961~ 1998-09-22 Step B: l-(Triphenylmethyl)-lH-imidazol-4-ylacetic acid methyl ester To a solution of the product from Step A (24.85g, 0.141mol) in DMF (l lSml) was added triethylamine (57.2 ml, 5 0.412mol) and triphenylmethyl bromide (55.3g, 0.171mol) and the suspension was stirred for 24 hrs. After this time, the reaction mixture was diluted with EtOAc and water. The organic phase was washed with saturated aqueous NaHCO3, dried (Na2SO4) and the solvent evaporated in vacuo. The residue was purified by chromatography (sio2~ gradient 10 elution, 0-100% EtOAc in hexanes; ) to provide the title compound as a white solid.
IH NMR CDC13, ~ 7.35(1H, s)~ 7.31(9H, m), 7.22(6H, m), 6.76(1H, s), 3.68(3H, s) and 3.60(2H, s) ppm.
~5 Step C: [1 -(4-Cyanobenzyl)- 1 H-imidazol-5-yl]acetic acid methyl ester.
To a solution of the product from Step B (8.00g, 20.9mmol) in acetonitrile (70 ml) was added 4-cyanobenzyl bromide (4.10g, 20.92 mmol) and heated at 55~C for 3 hr. After this time, the 20 reaction was cooled to room temperature and the resulting imidazolium salt was collected by filtration. The filtrate was heated at 55~C for 18hrs. The reaction mixture was cooled to room temperature and evaporated in vacuo. To the residue was added EtOAc (70 ml) and the resulting precipitate collected by filtration. The precipitated 25 imidazolium salt~s were combined, suspended in methanol (100 ml) and heated to reflux for 30 min. After thi.s time, the solvent wa~s removed in vac~o,. The resulting residue was suspended in EtOAc (75ml) and the solid isolated by filtration and washed with EtOAc.
The solid was treated with saturated aqueous NaHCO3 solution 30 (300ml) and CH2C12 (300ml) and stirred at room temperature for 2 hrs. The organic layer was separated, dried (MgSO4) and evaporated in vacuo to afford the title compound as a white solid lHNMR CDC13, ~ 7.65(1H, d, J=8Hz), 7.53(1H, s), 7.15(1H, d.
J=8Hz), 7.04(1H, s), 5.24(2H, s), 3.62(3H, s) and 3.45(2H, s) ppm.
CA 0224961~ 1998-09-22 W O 97/3~876 PCTrUS97/06257 Step D: 5-11-(4-cyanobenzyl)-lH-imidazolyllethanol.
To a stirred solution of the ester from example step C, (l.SOg, 5.8~mrnol), in methanol (20 ml) at 0~C, was added sodium 5 borohydride (l.OOg, 26.3mmol) portionwise over 5 min. The reaction was stirred at 0~C for 1 hr and then at room temperature for an additional 1 hr. The reaction was quenched by the addition of saturated NH4CI solution and the methanol evaporated in vacuo.. The residue was partitioned between EtOAc and saturated NaHC03 solution and the 10 organic extracts dried, (MgS04) and evaporated in vacuo. The residue was purified by chromatography (sio27 gradient elution, 4 to 10%
methanol in methylene chloride) to afford the title compound as a white solid.
1 H NMR CDC13 ~ 7.64(2H, d, J=8.2Hz), 7.57(1 H, s), 7.11 (2H, d, 15 J=8.2Hz), 6.97(1H, s), 5.23(2H, s), 3.79(2H, t, J=6.2Hz), 2.66(2H, t, J=6.2Hz) ppm.
Step E: 5~ (4-Cyanobenzyl~-imidazolyl)ethylmethanesulfonate.
A solution of 5-[1-(4-cyanobenzyl)-lH-imidazolyl]ethanol 20 (0.500 g, 2.20 mmol) in methylene chloride (6 ml) at 0~C was treated with Hunig's base (0.460ml, 2.64mmol) and methanesulfonyl chloride (0.204ml, 2.64mmol). After 2 hrs, the reaction was quenched by addition of saturated NaHC03 solution (SOml) and the mixture extracted with methylene chloride (50ml), dried (MgS04) and the 25 solvent evaporated in vacuo. The title compound was used without furthur purification.
lH NMR CDC13 ~ 7.69 (lH, s) 7.66(2H, d, J=8.2Hz), 7.15 (2H, d, J=8.2Hz), 7.04(1H, s), 5.24(2H, s), 4.31(2H, t, J=6.7Hz), 2.96(3H, s), and 2.88(2H, t, J=6.6Hz)ppm.
Step F~ [1 -(4-Cyanobenzyl)- 1 H-imidazol-5 -yl]ethyl ) -4-phenyl - imidazole bis hydrochloride salt.
To a suspension of sodium hydride (14.2mg, 60%
dispersion in mineral oil, 0.356mmol) in DMF (0.30 ml) at 0~C was CA 0224961~ 1998-09-22 W O 97/36876 PCTrUS97/06257 added 4-phenyl imidazole (4~.~mg, 0.339mmol), and stirred for 20 mins. A solution of the mesylate from step E (lOOmg, 0.339mmol) in DMF (0.50ml) was added to the Isolution and stirring continued at 0~C
for 1 hr and then at room temperature for 16 hrs. The reaction was 5 quenched with saturated ammonium chloride solution (O.lOml). and the the solvent evaporated in vacuo. The re~sidue was purified by chromatography (sio2~ gradient elution, 2-5% ammonium hydoxide:
acetonitrile. The resulting material was converted to the HCI salt by treating an EtOAc solution of the imidazole with gasseouls HCI and 10 evaporating the ~solvent in vacuo.
.. . . . . . .. . . .. . .. .
CA 0224961~ 1998-09-22 Anal. Calcd for C22H 19N5-2-00HCl- 1 -50H20:
C, Sg.29; H, 5.34; N, 15.45.
Found: C, 58.24; H, 5.47; N, 15.4~.
FAB HRMS exact mass calcd for C22H20N5 354.171 g71 (MH+); found 5 354.171948.
lH NMR CD30D ~ ~.93 (lH, s), 8.75(1H, s), 7.~6(1H, s), 7.76(2H, d, J=7.9Hz), 7.69(2H, d, 7.1Hz), 7.65-7.35(6H, m), 5.61(2H, s) and 4.53(2H,m)ppm.
10 In vitro inhibition of ras farne,syl transferase Assays offarnesyl-protein transferase.
Partially purified bovine FPTase and Ras peptides (Ras-CVLS, Ras-CVIM and Ras-CAIL) were prepared as described by Schaber et ah, J. Biol. Chem. 265:14701-14704 (1990), Pompliano, 15 et al., Biochemistry 31:3800 (1992) and Gibbs et al., PNAS U.S.A.
~6:6630-6634 (1989), respectively. Bovine FPTa.se was assayed in a volume of 100 ~1 containing 100 mM N-(2-hydroxy ethyl) piperazine-N'-(2-ethane sulfonic acid) (HEPl~S), pH 7.4, 5 mM MgC12, 5 mM
dithiothreitol (DTT), 100 mM [3H]-farnesyl diphosphate ([3H]-FPP; 740 20 CBq/mmol, New England Nuclear), 650 nM Ras-CVLS and 10 ~g/ml FPTase at 31 ~C for 60 min. 3~eactions were initiated with FPTase and stopped with 1 ml of 1.0 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 ,~-plate counter. The assay 25 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 radioactivity in the 30 presence of the test compound when compared to the amount of incorporation in the absence of the test compound.
- Human FPTa~se was prepared as described by Omer et al., Biochemistry 32:5167-5176 (1993). Human FPTase activity was assayed as described above with the exception that 0.1 % (w/v) CA 0224961~ 1998-09-22 W O 97/36876 PCTnUS97/06257 polyethylene glycol 20,000, 10 ~lM ZnC12 and 100 nM Ras-CVIM were added to the reaction mixture. Reactions were performed for 30 min., stopped with 100 ~1 of 30% (v/v) trichloroacetic acid (TCA) in ethanol and processed as described above for the bovine enzyme.
S The compounds of the instant invention described in the above Examples 1-7 were tested for inhibitory activity against human FPTase by the assay described above and were found to have IC50 of 50 ~lM.
In vivo ras farnesylation assay The cell line u~ed in this assay is a v-ras line derived from either Ratl or NIH3T3 cells, which expressed viral Ha-ras p21.
The assay is performed essentially as described in DeClue, J.E. et ah, Cancer Research 51 :712-717, (1991). Cells 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 labelled in 3 ml methionine-free DMEM supple-meted with 10% regular DMEM, 2~o fetal bovine serum and 400 mCi[35S]methionine (1000 Ci/mmol). After an additional 20 hours, the cells are lysed 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 count.s are bought to 1 ml with IP buffer (lysis buffer lacking DTT) and immunoprecipitated with the ra~s-specific monoclonal antibody Y13-259 (Furth, M.E. et ah, J. Virol. 43:294-304, (19X2)). 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/l % 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 autoradiographed. The intensities of the bands corresponding to CA 0224961~ 1998-09-22 W O 97/36876 PCT~US97/06257 farne~ylated and nonfarnesylated ras proteins are compared to determine the percent inhibition of farnesyl transfer to protein.
In viv(~ ~rowth inhibition assay To determine the biological consequences of FPTase inhibition, the effect of the compounds of the instant invention on the anchorage-independent growth of Ratl cells transformed with either a v-ras, v-7~af, or v-mos oncogene is tested. Cells transformed 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 are seeded at a density 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 appropriate concentration of the instant compound (dissolved 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 (38)
1. A compound represented by formula I:
or a pharmaceutically acceptable salt thereof, wherein:
R1a, R1b and R2 are independently selected from the group consisting of: hydrogen, aryl, substituted aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(O)m- wherein m is 0, 1 or 2, R8C(O)NR8-, CN, NO2, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(O)NR8- and C1-C6 alkyl, unsubstituted or substituted by 1-3 groups selected from the group consisting of:
halo, aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(O)m-, R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2 and R9OC(O)NR8-;
R3 and R4 are independently selected from the group consisting of: H, F, Cl, Br, -NR8 2, CF3, NO2, R8O-, R9S(O)m-, R8C(O)NH-, H2NC(NH)-, R8C(O)-, R8OC(O)-, N3, CN, R9OC(O)NR8-, C1-C20 alkyl, substituted or unsubstituted aryl and substituted or unsubstituted heterocyclyl;
A3 is selected from: -C~C~ , ~R8C=CR8~ , aryl, heteroaryl, -C(O)- or a bond;
provided that when A3 is heteroaryl, attachment of A3 the remainder of the molecule is through substitutable heteroaryl ring carbons;
X represents aryl or heteroaryl;
provided that when X is heteroaryl, attachment of X the remainder of the molecule is through substitutable heteroaryl ring carbons;
R6 is independently selected from the group consisting of:
hydrogen, aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-6 perfluoroalkyl, F, Cl, Br, R8O-, R9S(O)m-, R8C(O)NR8, CN, N02, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(O)NR8- and C1-C6 alkyl unsubstituted or substituted by 1-3 groups selected from: aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R8O-, R9S(O)m-, R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2 and R9OC(O)NR8-;
R7 is independently selected from the group consisting of:
hydrogen, aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-6 perfluoroalkyl, F, Cl, Br, R9O-, R9S(O)m-, R8C(O)NR8, CN, NO2, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(O)NR8- and C1-C6 alkyl unsubstituted or substituted by 1-3 groups selected from: aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R8O-, R9S(O)m-, R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2 and R9OC(O)NR8-;
each R8 is independently selected from hydrogen, C1-C6 alkyl, aryl and aralkyl;
each R9 is independently selected from C1-C6 alkyl and aryl;
A1 and A2 are independently selected from the group consisting of: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR8-, -NR8C(O)-, -O-, -N(R8)-, -S(O)2N(R8)-, -N(R8)S(O)2-, and S(O)m;
V is selected from the group consisting of: hydrogen, heterocyclyl, aryl, C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond, n is 0 and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R8 and to A1 is through a substitutable ring carbon;
W represents heterocyclyl;
each n and p independently represents 0, 1, 2, 3 or 4;
r is 0 to 5, provided that r is 0 when V is hydrogen, and t is 1.
or a pharmaceutically acceptable salt thereof, wherein:
R1a, R1b and R2 are independently selected from the group consisting of: hydrogen, aryl, substituted aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(O)m- wherein m is 0, 1 or 2, R8C(O)NR8-, CN, NO2, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(O)NR8- and C1-C6 alkyl, unsubstituted or substituted by 1-3 groups selected from the group consisting of:
halo, aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R8O-, R9S(O)m-, R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2 and R9OC(O)NR8-;
R3 and R4 are independently selected from the group consisting of: H, F, Cl, Br, -NR8 2, CF3, NO2, R8O-, R9S(O)m-, R8C(O)NH-, H2NC(NH)-, R8C(O)-, R8OC(O)-, N3, CN, R9OC(O)NR8-, C1-C20 alkyl, substituted or unsubstituted aryl and substituted or unsubstituted heterocyclyl;
A3 is selected from: -C~C~ , ~R8C=CR8~ , aryl, heteroaryl, -C(O)- or a bond;
provided that when A3 is heteroaryl, attachment of A3 the remainder of the molecule is through substitutable heteroaryl ring carbons;
X represents aryl or heteroaryl;
provided that when X is heteroaryl, attachment of X the remainder of the molecule is through substitutable heteroaryl ring carbons;
R6 is independently selected from the group consisting of:
hydrogen, aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-6 perfluoroalkyl, F, Cl, Br, R8O-, R9S(O)m-, R8C(O)NR8, CN, N02, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(O)NR8- and C1-C6 alkyl unsubstituted or substituted by 1-3 groups selected from: aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R8O-, R9S(O)m-, R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2 and R9OC(O)NR8-;
R7 is independently selected from the group consisting of:
hydrogen, aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-6 perfluoroalkyl, F, Cl, Br, R9O-, R9S(O)m-, R8C(O)NR8, CN, NO2, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2, R9OC(O)NR8- and C1-C6 alkyl unsubstituted or substituted by 1-3 groups selected from: aryl, heterocyclyl, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, perfluoroalkyl, F, Cl, Br, R8O-, R9S(O)m-, R8C(O)NR8-, CN, (R8)2NC(NR8)-, R8C(O)-, R8OC(O)-, N3, -N(R8)2 and R9OC(O)NR8-;
each R8 is independently selected from hydrogen, C1-C6 alkyl, aryl and aralkyl;
each R9 is independently selected from C1-C6 alkyl and aryl;
A1 and A2 are independently selected from the group consisting of: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR8-, -NR8C(O)-, -O-, -N(R8)-, -S(O)2N(R8)-, -N(R8)S(O)2-, and S(O)m;
V is selected from the group consisting of: hydrogen, heterocyclyl, aryl, C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond, n is 0 and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R8 and to A1 is through a substitutable ring carbon;
W represents heterocyclyl;
each n and p independently represents 0, 1, 2, 3 or 4;
r is 0 to 5, provided that r is 0 when V is hydrogen, and t is 1.
2. A compound in accordance with claim 1 wherein R1a ,R1b and R2 are independently selected from: hydrogen, -N(R8)2, R8C(O)NR8- or unsubstituted or substituted C1-C6 alkyl wherein the substituent on the substituted C1-C6 alkyl is selected from unsubstituted or substituted aryl, -N(R8)2, R8O- and R8C(O)NR8-.
3. A compound in accordance with claim 1 wherein R3 and R4 are selected from: hydrogen, C1-C6 alkyl, Cl, Br, F, R8O- and CF3.
4. A compound in accordance with claim 1 wherein A3 represents -C~C-, -CR8=CR8-, -C(O)- or a bond..
5. A compound in accordance with claim 1 wherein A3 represents -C(O)- .
6. A compound in accordance with claim 1 wherein A3 represents aryl or heteroaryl.
7. A compound in accordance with claim 1 wherein R6 represents CN.
8. A compound in accordance with claim 1 wherein R7 represents hydrogen, unsubstituted or substituted C1-C6 alkyl.
9. A compound in accordance with claim 1 wherein R8 represents H or C1-6 alkyl, and R9 is C1-6 alkyl.
10. A compound in accordance with claim 1 wherein A1 and A2 are independently selected from: a bond, -C(O)NR8-, -NR8C(O)-, -O-, -N(R8)-, -S(O)2N(R8)- and-N(R8)S(O)2-.
11. A compound in accordance with claim 1 wherein V
is selected from hydrogen, heterocyclyl and aryl.
is selected from hydrogen, heterocyclyl and aryl.
12. A compound in accordance with claim 11 wherein V
is phenyl.
is phenyl.
13. A compound in accordance with claim 1 wherein W
is heterocyclyl selected from imidazolinyl, imidazolyl, oxazolyl, pyrazolyl, pyyrolidinyl, thiazolyl and pyridyl.
is heterocyclyl selected from imidazolinyl, imidazolyl, oxazolyl, pyrazolyl, pyyrolidinyl, thiazolyl and pyridyl.
14. A compound in accordance with claim 1 wherein W
is selected from imidazolyl and pyridyl.
is selected from imidazolyl and pyridyl.
15. A compound in accordance with claim 1 wherein X
represents aryl.
represents aryl.
16. A compound in accordance with claim 15 wherein X
represents phenyl.
represents phenyl.
17. A compound in accordance with claim 1 wherein X
represents heteroaryl.
represents heteroaryl.
18. A compound in accordance with claim 17 wherein X
represents pyridyl.
represents pyridyl.
19. A compound in accordance with claim 1 wherein m is 0 or 2.
20. A compound in accordance with claim 1 wherein n and p are 0, 1, 2 or 3.
21. A compound in accordance with claim 1 wherein t is 1.
22. A compound in accordance with claim 1 represented by formula Ia:
wherein:
R3, R4, A3, R8, R9, X, m, n, p and r are as originally defined;
each R1a and R2 is independently selected from hydrogen and C1-C6 alkyl;
each R1b is independently selected from: hydrogen, aryl, heterocyclyl, C3-10 cycloalkyl, C2-6 alkenyl, R8O-, -N(R8)2 and C1-C6 alkyl unsubstituted or substituted by aryl, heterocyclyl, cycloalkyl, alkenyl, R8O- and -N(R8)2;
R6 is independently selected from: hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R8O-, R8C(O)NR8-, CN, NO2, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2, or R9OC(O)NR8-, and C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R8O-, R8C(O)NR8-, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2 and R9OC(O)NR8-;
R7 represents H or C1-6 alkyl;
A1 and A2 are independently selected from: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR8-, O, -N(R8)- and S(O)m;
and V is selected from: hydrogen; aryl; heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl and thienyl; C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a a heteroatom selected from O,S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R8 and to A1 is through a substitutable ring carbon.
wherein:
R3, R4, A3, R8, R9, X, m, n, p and r are as originally defined;
each R1a and R2 is independently selected from hydrogen and C1-C6 alkyl;
each R1b is independently selected from: hydrogen, aryl, heterocyclyl, C3-10 cycloalkyl, C2-6 alkenyl, R8O-, -N(R8)2 and C1-C6 alkyl unsubstituted or substituted by aryl, heterocyclyl, cycloalkyl, alkenyl, R8O- and -N(R8)2;
R6 is independently selected from: hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R8O-, R8C(O)NR8-, CN, NO2, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2, or R9OC(O)NR8-, and C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R8O-, R8C(O)NR8-, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2 and R9OC(O)NR8-;
R7 represents H or C1-6 alkyl;
A1 and A2 are independently selected from: a bond, -CH=CH-, -C~C-, -C(O)-, -C(O)NR8-, O, -N(R8)- and S(O)m;
and V is selected from: hydrogen; aryl; heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl and thienyl; C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a a heteroatom selected from O,S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R8 and to A1 is through a substitutable ring carbon.
23. A compound in accordance with claim 1 represented by formula Ib:
wherein:
R1a R1b, R2, A1, A2, R3, R4, R6, R8, R9, X, m, n, p and r are as originally defined;
R7 is selected from: hydrogen and C1-C6 alkyl;
V is selected from: hydrogen, heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl and thienyl; aryl; C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond, n is 0 and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R8 and to A1 is through a substitutable ring carbon;
and W represents heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl and isoquinolinyl.
wherein:
R1a R1b, R2, A1, A2, R3, R4, R6, R8, R9, X, m, n, p and r are as originally defined;
R7 is selected from: hydrogen and C1-C6 alkyl;
V is selected from: hydrogen, heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl and thienyl; aryl; C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond, n is 0 and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R8 and to A1 is through a substitutable ring carbon;
and W represents heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl and isoquinolinyl.
24. A compound in accordance with claim 1 represented by formula Ic:
wherein:
R1a, R1b, R2, A1, A2, R3, R4, R6, R8, R9, X, m, n, p and r are as originally defined;
R7 is selected from: hydrogen and C1-C6 alkyl;
V is selected from: hydrogen, heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, aryl, C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond, n is 0 and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R8 and to A1 is through a substitutable ring carbon;
and W represents heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl and isoquinolinyl.
wherein:
R1a, R1b, R2, A1, A2, R3, R4, R6, R8, R9, X, m, n, p and r are as originally defined;
R7 is selected from: hydrogen and C1-C6 alkyl;
V is selected from: hydrogen, heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl, isoquinolinyl, and thienyl, aryl, C1-C20 alkyl wherein from 0 to 4 carbon atoms are replaced with a heteroatom selected from O, S, and N, and C2-C20 alkenyl, provided that V is not hydrogen if A1 is S(O)m and V is not hydrogen if A1 is a bond, n is 0 and A2 is S(O)m;
provided that when V is heterocycle, attachment of V to R8 and to A1 is through a substitutable ring carbon;
and W represents heterocyclyl selected from pyrrolidinyl, imidazolyl, pyridinyl, thiazolyl, indolyl, quinolinyl and isoquinolinyl.
25. A compound in accordance with claim 1 represented by formula Id:
wherein:
each R2 is independently selected from hydrogen and C1-C6 alkyl, R3, R4, A3, R8, R9, X, m and p are as originally defined;
and R6 is selected from the group consisting of:
hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R8O-, R8C(O)NR8-, CN, NO2, (R8)2N-C(NR8), R8C(O)-, R8OC(O)-, -N(R8)2, or R9OC(O)NR8- and C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R8O-, R8C(O)NR8-, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2 or R9OC(O)NR8-.
wherein:
each R2 is independently selected from hydrogen and C1-C6 alkyl, R3, R4, A3, R8, R9, X, m and p are as originally defined;
and R6 is selected from the group consisting of:
hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R8O-, R8C(O)NR8-, CN, NO2, (R8)2N-C(NR8), R8C(O)-, R8OC(O)-, -N(R8)2, or R9OC(O)NR8- and C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R8O-, R8C(O)NR8-, (R8)2N-C(NR8)-, R8C(O)-, R8OC(O)-, -N(R8)2 or R9OC(O)NR8-.
26. A compound in accordance with claim 1 represented by formula Ie:
wherein:
X and A3 are as originally defined;
each R2 is independently selected from: hydrogen and C1-C6 alkyl;
R3 and R4 are independently selected from H, F, Cl, Br, N(R8)2, CF3, NO2, (R8)O-, (R9)S(O)m-, (R8)C(O)NH-, H2N-C(NH)-, (R8)C(O)-, (R8)OC(O)-, N3, CN, (R9)OC(O)NR8-, C1-C20 alkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclyl;
and R8, R9, m and p are as originally defined.
wherein:
X and A3 are as originally defined;
each R2 is independently selected from: hydrogen and C1-C6 alkyl;
R3 and R4 are independently selected from H, F, Cl, Br, N(R8)2, CF3, NO2, (R8)O-, (R9)S(O)m-, (R8)C(O)NH-, H2N-C(NH)-, (R8)C(O)-, (R8)OC(O)-, N3, CN, (R9)OC(O)NR8-, C1-C20 alkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heterocyclyl;
and R8, R9, m and p are as originally defined.
27. A compound in accordance with claim 1 represented by the formula:
or or a pharmaceutically acceptable salt thereof.
or or a pharmaceutically acceptable salt thereof.
28. A pharmaceutical composition comprising a compound in accordance with claim 1 in combination with a pharmaceutically acceptable carrier.
29. A method of inhibiting farnesyl-protein transferase in a mammalian patient in need of such treatment which comprises administering to said patient an effective amount of a compound in accordance with claim 1.
30. A method of treating cancer in a mammalian patient in need of such treatment which comprises administering to said patient an anti-cancer effective amount of a compound in accordance with claim 1.
31. A method for treating neurofibromin benign proliferative disorder in a mammalian patient in need of such treatment which comprises administering to said patient an effective amount of a compound in accordance with claim 1 to treat neurofibromin benign proliferative disorder.
32. A method for treating blindness related to retinal vascularization in a mammalian patient in need of such treatment which comprises administering to said patient an effective amount of a compound in accordance with claim 1 to treat blindness related to retinal vascularization.
33. A method for treating infections from hepatitis delta and related viruses in a mammalian patient in need of such treatment which comprises administering to said patient an anti-viral effective amount of a compound in accordance with claim 1.
34. A method for preventing restenosis in a mammalian patient in need of such treatment which comprises administering to said patient a compound in accordance with claim 1 in an amount effective for preventing restenosis.
35. A method for treating polycystic kidney disease in a mammalian patient in need of such treatment which comprises administering to said patient a compound in accordance with claim 1 in an amount effective to treat polycystic kidney disease.
36. A method for treating or preventing a disease selected from cancer, neurofibromin benign proliferative disorder, blindness related to retinal vascularization, infections from hepatitis delta and related viruses, restenosis and polycystic kidney disease in a mammalian patient in need of such treatment, which comprises administering to an effective amount of a compound in accordance with claim 1 to treat or prevent said disease.
37. A pharmaceutical composition made by combining the compound of Claim 1 and a pharmaceutically acceptable carrier.
38. A process for making a pharmaceutical composition comprising combining a compound of Claim 1 and a pharmaceutically acceptable carrier.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US1459296P | 1996-04-03 | 1996-04-03 | |
| US60/014,592 | 1996-04-03 | ||
| GBGB9613462.2A GB9613462D0 (en) | 1996-06-27 | 1996-06-27 | Inhibitors of farnesyl-protein transferase |
| GB9613462.2 | 1996-06-27 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2249615A1 true CA2249615A1 (en) | 1997-10-09 |
Family
ID=26309577
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002249615A Abandoned CA2249615A1 (en) | 1996-04-03 | 1997-04-01 | Inhibitors of farnesyl-protein transferase |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0891334A1 (en) |
| JP (1) | JP2000507955A (en) |
| AU (1) | AU715658B2 (en) |
| CA (1) | CA2249615A1 (en) |
| WO (1) | WO1997036876A1 (en) |
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| KR100375421B1 (en) * | 1997-07-15 | 2003-05-12 | 주식회사 엘지생명과학 | Farnesyl transferase inhibitor having anti-tumor properties |
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| EP2073802A1 (en) | 2006-10-12 | 2009-07-01 | Astex Therapeutics Limited | Pharmaceutical combinations |
| US8916552B2 (en) | 2006-10-12 | 2014-12-23 | Astex Therapeutics Limited | Pharmaceutical combinations |
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| EP2073807A1 (en) | 2006-10-12 | 2009-07-01 | Astex Therapeutics Limited | Pharmaceutical combinations |
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| KR20150066608A (en) | 2007-12-07 | 2015-06-16 | 버텍스 파마슈티칼스 인코포레이티드 | Processes for producing cycloalkylcarboxiamido-pyridine benzoic acids |
| NZ736561A (en) | 2008-02-28 | 2018-02-23 | Vertex Pharma | Heteroaryl derivatives as cftr modulators |
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| UA103319C2 (en) | 2008-05-06 | 2013-10-10 | Глаксосмитклайн Ллк | Thiazole- and oxazole-benzene sulfonamide compounds |
| US9156812B2 (en) | 2008-06-04 | 2015-10-13 | Bristol-Myers Squibb Company | Crystalline form of 6-[(4S)-2-methyl-4-(2-naphthyl)-1,2,3,4-tetrahydroisoquinolin-7-yl]pyridazin-3-amine |
| EP2375899B1 (en) | 2009-01-12 | 2015-02-25 | Array Biopharma Inc. | Piperidine-containing compounds and use thereof in the treatment of diabetes |
| TW201100398A (en) | 2009-03-31 | 2011-01-01 | Arqule Inc | Substituted indolo-pyridinone compounds |
| CN102638982B (en) | 2009-05-12 | 2015-07-08 | 百时美施贵宝公司 | Crystalline forms of (s)-7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorohphenyl)-1,2,3,4-tetrahydroisoquinoline and use thereof |
| AU2010247849B2 (en) | 2009-05-12 | 2015-11-19 | Albany Molecular Research, Inc. | Aryl, heteroaryl, and heterocycle substituted tetrahydroisoquinolines and use thereof |
| JP2013523833A (en) | 2010-04-07 | 2013-06-17 | バーテックス ファーマシューティカルズ インコーポレイテッド | 3- (6- (1- (2,2-difluorobenzo [D] [1,3] dioxol-5-yl) cyclopropanecarboxamido) -3-methylpyridin-2-yl) benzoic acid pharmaceutical composition and Its administration |
| CN105848657B (en) | 2013-11-12 | 2020-05-22 | 沃泰克斯药物股份有限公司 | Methods of making pharmaceutical compositions for treating CFTR mediated diseases |
| PL3221692T3 (en) | 2014-11-18 | 2021-12-06 | Vertex Pharmaceuticals Inc. | Process of conducting high throughput testing high performance liquid chromatography |
| KR20200143376A (en) | 2018-03-13 | 2020-12-23 | 샤이어 휴먼 지네틱 테라피즈 인크. | Substituted imidazopyridines and uses thereof as plasma kallikrein inhibitors |
| JP2022549601A (en) | 2019-09-18 | 2022-11-28 | 武田薬品工業株式会社 | heteroaryl plasma kallikrein inhibitors |
| FI4031547T3 (en) | 2019-09-18 | 2024-09-16 | Takeda Pharmaceuticals Co | Plasma kallikrein inhibitors and uses thereof |
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| US4837333A (en) * | 1987-05-14 | 1989-06-06 | G. D. Searle & Co. | Substituted alkylidene imidazoles |
| DE3811574A1 (en) * | 1988-03-31 | 1989-10-19 | Schering Ag | N-SUBSTITUTED IMIDAZOLES, METHODS FOR THEIR PRODUCTION AND THEIR USE IN MEDICINAL PRODUCTS |
| DE3813885A1 (en) | 1988-04-20 | 1989-11-02 | Schering Ag | 1-CHLORPYRIMIDINYL-1H-1,2,4-TRIAZOL-3-SULPHONIC ACID AMIDE, METHOD FOR THE PRODUCTION THEREOF AND THEIR USE AS AGENT WITH HERBICIDES, FUNGICIDES AND PLANT GROWTH REGULATING EFFECT |
| FR2657610A1 (en) | 1990-01-29 | 1991-08-02 | Rhone Poulenc Agrochimie | TRIAZOLOPYRIDINES HERBICIDES. |
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| JPH05201974A (en) | 1991-11-25 | 1993-08-10 | Sumitomo Chem Co Ltd | Heterocyclic compound and herbicide containing the compound as active component |
| AU5285593A (en) * | 1992-10-21 | 1994-05-09 | Sankyo Company Limited | Azole compound |
| GB2273704B (en) * | 1992-12-16 | 1997-01-22 | Orion Yhtymae Oy | Triazolyl diaryl selective aromatase inhibiting compounds |
| CZ104795A3 (en) | 1994-04-29 | 1996-02-14 | Lilly Co Eli | Benzimidazole derivative, process of its preparation, its use for preparing a pharmaceutical preparation and the pharmaceutical composition containing thereof |
-
1997
- 1997-04-01 CA CA002249615A patent/CA2249615A1/en not_active Abandoned
- 1997-04-01 EP EP97918643A patent/EP0891334A1/en not_active Withdrawn
- 1997-04-01 JP JP9535634A patent/JP2000507955A/en active Pending
- 1997-04-01 AU AU26702/97A patent/AU715658B2/en not_active Ceased
- 1997-04-01 WO PCT/US1997/006257 patent/WO1997036876A1/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| AU715658B2 (en) | 2000-02-10 |
| WO1997036876A1 (en) | 1997-10-09 |
| JP2000507955A (en) | 2000-06-27 |
| EP0891334A1 (en) | 1999-01-20 |
| AU2670297A (en) | 1997-10-22 |
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