WO1996031505A1 - Tricyclic compounds useful for inhibition of g-protein function and for treatment of proliferative diseases - Google Patents

Abstract

Novel compounds of Formula (1.0) are disclosed. Also disclosed is a method of inhibiting Ras function and therefore inhibiting the abnormal growth of cells. The method comprises administering a compound of the formula to a biological system. In particular, the method inhibits the abnormal growth of cells in a mammal such as a human being.

Description

TRICYCLIC COMPOUNDS USEFUL FOR INHIBITION OF G-PROTEIN FUNCTION AND FOR TREATMENT OF PRQLIFERATIVE DISEASES

BACKGROUND

International Publication Number WO92/11034, published July 9, 1992, discloses a method of increasing the sensitivity of a tumor to an antineoplastic agent, which tumor is resistant to the antineoplastic agent, by the concurrent administration of the antineoplastic agent and a potentiating agent of the formula:

wherein Y is hydrogen, substituted carboxylate or substituted sulfonyl. Examples of such potentiating agents include 1 1-(4-piperidylidene)-5H- benzo[5,6]cyclohepta[1 ,2-b]pyridines such as Loratadine.

To acquire transforming potential, the precursor of the Ras oncoprotein must undergo farnesylation of the cysteine residue located in a carboxyl-terminal tetrapeptide. Inhibitors of the enzyme that catalyzes this modification, farnesyl protein transferase, have therefore been suggested as anticancer agents for tumors in which Ras contributes to transformation. Mutated, oncogenic forms of ras are frequently found in many human cancers, most notably in more than 50% of colon and pancreatic carcinomas (Kohl et al., Science, Vol. 260, 1834 to 1837, 1993).

A welcome contribution to the art would be compounds useful for the inhibition of farnesyl protein transferase. Such a contribution is provided by this invention. SUMMARY OF THE INVENTION Inhibition of farnesyl protein transferase by tricyclic compounds of this invention has not been reported previously. Thus, this invention provides a method for inhibiting farnesyl protein transferase using tricyclic compounds of this invention which: (i) potently inhibit farnesyl protein transferase, but not geranylgeranyl protein transferase I, in vitro: (ii) block the phenotypic change induced by a form of transforming Ras which is a farnesyl acceptor but not by a form of transforming Ras engineered to be a geranylgeranyl acceptor; (iii) block intracellular processing of Ras which is a farnesyl acceptor but not of Ras engineered to be a geranylgeranyl acceptor; and (iv) block abnormal cell growth in culture induced by transforming Ras.

This invention provides a method for inhibiting the abnormal growth of cells, including transformed cells, by administering an effective amount of a compound of this invention. Abnormal growth of cells refers to cell growth independent of normal regulatory mechanisms (e.g., loss of contact inhibition). This includes the abnormal growth of: (1) tumor cells (tumors) expressing an activated Ras oncogene; (2) tumor cells in which the Ras protein is activated as a result of oncogenic mutation in another gene; and (3) benign and malignant cells of other proliferative diseases in which aberrant Ras activation occurs.

The compounds useful in the claimed methods are novel compounds represented by Formula (1.0)

wherein:

A and B are independently selected from H, halo or C-ι-C-₆ alkyl; Z is N or CH;

W is CH, CH2, O or S, wherein the dotted line to W represents a double bond which is present when W is CH; R¹ is selected from the group consisting of:

wherein W, A and B are as defined above; or

R¹ is a group D, wherein D is -C(0)-(CH₂)_s-R⁵. -C(0)0-(CH₂)_m-R⁵ or -C(0)NH-(CH2)m-R⁵. wherein R⁵ is aryl, (such as phenyl, B-substituted phenyl wherein B is as defined below), heterparyl, (such as pyridyl or pyridyl N-oxide), heterocycloalkyl, or a group of the formula

wherein g = 1 or 2, and R¹¹ represents H, C-i-Cβ alkyl, haloalkyi or -C(O)- R⁹ wherein R⁹ is C C₆ alkyl, C Cβ alkoxy or -NH(R^10A) wherein R¹°A j_S H or alkyl, or the group -C(0)-R⁹ represents an acyl radical of a naturally occurring amino acid; or

R¹ is a group of the formula:

wherein: :

—

-S0₂-, or a single bond;

(b) x is 0, 1, 2, 3, 4, 5 or 6;

(c) each R^a and each R^b is independently selected from H, aryl, alkyl, alkoxy, aralkyi, amino, alkylamino, heterocyloalkyl, -COOR⁶⁰,

-NH CfC JzR⁶⁰ (wherein z is 0 or 1), or -(CH)_wS(0)_tR⁶⁰ (wherein w is 0, 1 , 2 or 3, and t is 0, 1 or 2); or R^a and R^b taken together can represent cycloalkyl, =N-0-alkyl, =0 or heterocycloalkyi; with the proviso that for the same carbon, R^a is not selected from alkoxy, amino, alkylamino or -NH{C(0)}_zR⁶⁰ when R^b is selected from alkoxy, amino, alkylamino or -NH{C(0)}_zR⁶⁰; and with the proviso that when T is a single bond, for the first carbon containing R^a and R^b, R^a and R^b are not selected from alkoxy, alkylamino, amino or -NHR⁶⁰ (i.e., -NH{C(0)}_zR⁶⁰ wherein z is 0) (i.e., R^a and R^b on the first carbon bound to T, when T is a single bond, are not alkoxy, alkylamino, amino or -NHR⁶⁰); and

(d) R⁹² can represent H, alkyl, aryl, aryloxy, arylthio, aralkoxy, aralkyi, heteroaryl or heterocycloalkyi;

R⁶⁰ represents H, alkyl, aryl or aralkyi;

R is H or Cι-C6 alkyl; R2 is selected from: -C(0)OR⁶, -C(0)NR⁶R⁷, Cι-C₈ alkyl, C₂-C₈ alkenyl, C₂-Cβ alkynyl, substituted (Cι-Cβ)alkyl, substituted (C₂-C8)alkenyl, substituted (C₂-Cβ)alkynyl, wherein said substituted groups have one or more substituents selected from:

1 ) aryl, heteroaryl, heterocycloalkyi, B-substituted aryl, B-substituted heteroaryl or B-substituted heterocycloalkyi, wherein B is selected from C1-C4 alkyl, phenyl, -(CH₂)_nOR⁶ -(CH₂)_nNR⁶R⁷ and halo;

2) C₃-C₆ cycloalkyl;

3) -OR⁶;

4) -S(0)_tR⁶; 5) -NR⁶R⁷;

6) -N(R⁶)-C(0)R⁷;

7) -N(R⁶)-C(0)NR7R12_;

8) -0-C(0)NR⁶R⁷;

9) -0-C(0)OR⁶; 10) -S0₂NR⁶R⁷;

11 ) -N(R⁶)-S0₂-R⁷; 12) -CJOJNR^βR⁷;

13) -C(0)OR⁶; and provided that: where R¹ is D, R² is not H; where R¹ is D and R² is C-|-Cβ alkyl, the substituents on said alkyl group are not substituents 4), 5), 9) or 13); and where R¹ is D, and R² is C-i-Cβ alkyl substituted by the group -OR⁶, R⁶ is not H, alkyl, aryl, substituted aryl, aryl-substituted alkyl or nitro- phenylsubstituted alkyl;

R⁶, R⁷ and R¹² are independently selected from H, C1-C4 alkyl, (C3-C6)cycloalkyl, aryl, arylalkyl (i.e., aralkyi), heteroaryl, heteroarylalkyl, heterocycloalkyi, substituted (Cι-C4)alkyl, substituted (C3-C6)cycloalkyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalky or substituted heterocycloalkyi, wherein said substituted groups have one or more substituents (e.g., 1-3) selected from: C1-C4 alkoxy, aralkyi, heteroarylalkyl, -N0₂, (C3-C-ιo)alkoxyalkoxy (e.g., -0-(C-|- C4)alkyl-0-(Cι-C4)alkyl), (C3-C6)cycloalkyl (e.g., cyclopropyl or cyclohexyl), aryl, -CN, nitro-phenyl, methylenedioxyphenyl, heteroaryl, heterocycloalkyi, halo, -OH, -COOH, -C(0)R¹⁴, -C(0)OR¹⁴, -C(0)NR⁶R⁷ (e.g., -C(O)NR ⁰R¹⁵), -N(R⁶)C(0)R¹⁴, -S(0)_tR¹⁴ (e.g., -S-(Cι-C₄) and -S0₂R¹⁴) or -NR¹⁰R¹⁵; provided that R⁶, R⁷ and R¹² are not -CH₂OH or -CH₂NR¹⁰R¹⁵ when said R⁶, R⁷ or R¹² is directly bonded to a heteroatom, and further provided that R⁶ is not H for groups 4) and 9), and R⁷ is not H for group 6); optionally, when R⁶ and R⁷ are bound to the same nitrogen, R⁶ and R⁷ together with the nitrogen to which they are bound, form a 5 to 7 membered heterocycloalkyi ring which optionally contains 0, NR⁶ (e.g., NR⁸), or S(0)t (e.g., S) wherein t is 0, 1 or 2; optionally, when R⁷ and R¹² are bound to the same nitrogen, R⁷ and R¹² together with the nitrogen to which they are bound, form a 5 to 7 membered heterocycloalkyi ring which optionally contains O, NR⁶ (e.g., NR⁸), or S(0)t (e.g., S) wherein t is 0, 1 or 2;

R⁸, R¹⁰ and R¹⁵ are independently H, C1-C4 alkyl or arylalkyl; R¹⁴ is C1-C4 alkyl, aryl or arylalkyl; m = 0, 1, 2 or 3; n = 0, 1 , 2, 3 or 4; s = 1, 2 or 3; and t = 0, 1 or 2; or pharmaceutically acceptable salts thereof. This invention also provides a method for inhibiting tumor growth by administering an effective amount of the tricyclic compounds, described herein, to a mammal (e.g., a human) in need of such treatment. In particular, this invention provides a method for inhibiting the growth of tumors expressing an activated Ras oncogene by the administration of an effective amount of the above described compounds. Examples of tumors which may be inhibited include, but are not limited to, lung cancer (e.g., lung adenocarcinoma), pancreatic cancers (e.g., pancreatic carcinoma such as, for example, exocrine pancreatic carcinoma), colon cancers (e.g., colorectal carcinomas, such as, for example, colon adenocarcinoma and colon adenoma), myeloid leukemias (for example, acute myelogenous leukemia (AML)), thyroid follicular cancer, myelodysplastic syndrome (MDS), bladder carcinoma and epidermal carcinoma.

It is believed that this invention also provides a method for inhibiting proliferative diseases, both benign and malignant, wherein Ras proteins are aberrantly activated as a result of oncogenic mutation in other genes- i.e., the Ras gene itself is not activated by mutation to an oncogenic fornv- with said inhibition being accomplished by the administration of an effective amount of the tricyclic compounds described herein, to a mammal (e.g., a human) in need of such treatment. For example, the benign proliferative disorder neurofibromatosis, or tumors in which Ras is activated due to mutation or overexpression of tyrosine kinase oncogenes (e.g., neu, src, abl, lck, and fyn), may be inhibited by the tricyclic compounds described herein. The compounds of this invention inhibit farnesyl protein transferase and the famesylation of the oncogene protein Ras. This invention further provides a method of inhibiting ras farnesyl protein transferase, in mammals, especially humans, by the administration of an effective amount of the tricyclic compounds described above. The administration of the compounds of this invention to patients, to inhibit farnesyl protein transferase, is useful in the treatment of the cancers described above. The tricyclic compounds useful in the methods of this invention inhibit the abnormal growth of cells. Without wishing to be bound by theory, it is believed that these compounds may function through the inhibition of G-proteiπ function, such as ras p21 , by blocking G-protein isoprenylation, thus making them useful in the treatment of proliferative diseases such as tumor growth and cancer. Without wishing to be bound by theory, it is believed that these compounds inhibit ras farnesyl protein transferase, and thus show antiproliferative activity against ras transformed cells.

DETAILED DESCRIPTION OF THE INVENTION

All of the publications cited herein are hereby expressly incorporated in their entirety by reference.

As used herein, the following terms are used as defined below unless otherwise indicated:

"M^+" represents the molecular ion of the molecule in the mass spectrum; "MH+^* represents the molecular ion plus hydrogen of the molecule in the mass spectrum; "Bu" represents butyl; "Et" represents ethyl;

"Tr" represents trityl, (i.e., triphenylmethyl); "Me" represents methyl;

"Ph" represents phenyl; "TBS" represents t-butyldimethylsilyl; "BOC" represents t-butoxycarbonyl; "FMOC" represents 9-fluorenylmethoxycarbonyl; "alkyl" (including the alkyl portions of alkoxy, alkylamino and dialkylamino) represents straight and branched carbon chains and contains from one to twenty carbon atoms, preferably one to six carbon atoms; said alkyl group optionally being substitued with one, two or three groups independently selected from hydroxy, alkoxy, halo (e.g., -CF₃), amino, alkylamino, dialkylamino, N-acylalkylamino, N-alkyl-N-acylamino, or -S(0)t- alkyl (wherein t is 0, 1 or 2), and wherein the alkyl portion of said optional groups are as defined above;

"alkenyl" represents straight and branched carbon chains having at least one carbon to carbon double bond and containing from 2 to 12 carbon atoms, preferably from 2 to 6 carbon atoms and most preferably from 3 to 6 carbon atoms;

"alkynyl" represents straight and branched carbon chains having at least one carbon to carbon triple bond and containing from 2 to 12 carbon atoms, preferably from 2 to 6 carbon atoms; "aralkyi" represents an alkyl group, as defined above, wherein one or more hydrogen atoms of the alkyl moiety have been replaced by one or more aryl groups, as defined below (e.g., benzyl and diphenylmethyl); "aryl" (including the aryl portion of aryloxy and aralkyi) represents a monocyclic, bicyclic or tricyclic carbocyclic group containing from 6 to 15 carbon atoms and comprising at least one aromatic ring, such as phenyl, naphthyl, phenanthryl, tetrahydronaphthyl or indanyl, with all available substitutable carbon atoms of the carbocyclic group being intended as possible points of attachment, said carbocyclic group being optionally substituted with one or more, preferably 1 to 3, substituents, independently selected from: (1) halo, (2) alkyl (e.g., Ci to Cβ alkyl), (3) hydroxy, (4) alkoxy (e.g., Ci to Ce alkoxy), (5) -CN, (6) phenyl, (7) phenoxy, (8) -CF3, (9) amino, (10) alkylamino, (11) dialkylamino, (12) aryl, (13) aralkoxy, (14) aryloxy, (15) -S(0)t-aryl (wherein t is 0, 1, or 2), (16) -COOR⁶⁰ (R⁶⁰ is as defined above), (17) -N0₂, or (18) substituted C1-C₆ alkyl wherein said alkyl group is substituted with 1 , 2, or 3 groups independently selected from (a) amino, (b) alkylamino, (c) dialkylamino, (d) aryl, (e) N-acylalkyl-amino, (f) N-alkyl-N-acylamino, (g) N-aralkyl-N- acylamino, (h) hydroxy, (i) alkoxy, G) halo (e.g., CF₃), or (k) heterocycloalkyi, provided that when there are two or more hydroxy, amino, alkylamino or dialkylamino substituents on the substituted C-i-Cβ alkyl group, the substituents are on different carbon atoms; or alternatively said aryl group may be fused through adjacent atoms to form a fused ring containing up to four carbon and/or heteroatoms (e.g., methylene dioxyphenyl, indanyl, tetralinyl, dihydrobenzofuranyl);

"aralkoxy" - represents an aralkyi group, as defined above, in which the alkyl moiety is covalently bonded to an adjacent structural element through an oxygen atom, for example, benzyloxy;

"aryloxy" - represents an aryl group, as defined above, covalently bonded to an adjacent structural element through an oxygen atom, for example, phenoxy;

"arylthio" - represents an aryl group, as defined above, covalently bonded to an adjacent structural element through a sulfur atom, for example, phenylthio;

"cycloalkyl" represents a saturated or unsaturated nonaromatic carbocyclic ring of from 3 to 8 carbon atoms, preferably 3 to 6 carbon atoms; "halo" represents fluoro, chloro, bromo and iodo;

"heterocycloalkyi" represents a saturated or unsaturated nonaromatic carbocyclic ring containing from 3 to 15 carbon atoms, preferably from 4 to 6 carbon atoms, and from 1 to 3 heteroatoms selected from O, S, -SO2- or NR¹⁰ (suitable heterocycloalkyi groups include tetrahydrofuranyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothienyl, piperidinyl, pyrrolidinyl, piperazinyl, dioxanyl, morpholino, diaza-2,2,2- bicyclooctane etc.), wherein any of the available substitutable carbon and nitrogen atoms in the ring are optionally substituted with one, two, three or more groups independently selected from C-i-Cβ alkyl, aryl, aralkyi, haloalkyi, amino, alkylamino, dialkylamino, -S(0)t-aryl (wherein t is 0, 1 or 2 and aryl is defined above), -C(0)R⁹ (wherein R⁹ is defined above) or an acyl radical of a naturally occuring amino acid; "heteroaryl" (including the heteroaryl portion of heteroarylalkyl) represents a monocyclic, bicyclic or tricyclic group containing from 2 to 14 carbon atoms and comprising one or more, (preferably 1 to 3), heteroatoms selected from O, S or N, said heteroatoms interrupting a carbocyclic ring structure and having a sufficient number of delocalized pi electrons to provide aromatic character, such as triazolyl, pyridyl, imidazolyl, thienyl, furanyl, imidazolyl, quinolyl, isoquinolyl, benzofuranyl, benzopyranyl, benzothienyl, thiazolyl, indolyl, naphthyridinyl, or pyridyl N- oxide, wherein pyridyl N-oxide can be represented as:

with all available substitutable carbon and heteroatoms of the cyclic group being intended as possible points of attachment, said cyclic group being optionally substituted with 1 , 2, 3 or more groups independently selected from halo, alkyl, aryl, aralkyi, heteroaryl, hydroxy, alkoxy, phenoxy, -N0_2> -CF₃, amino, alkylamino, dialkylamino, and -COOR⁶⁰ wherein R⁶⁰ is as defined above (e.g., benzyl).

As used herein, the term "tertiary amine base" means DMAP, pyridine or a trialkylamine, such as Et₃N or Hϋnigs base;

"hydroxide base" means NH₄OH or an alkali metal or alkaline earth metal hydroxide, such as LiOH, NaOH, KOH, Mg(OH)₂ or Ca(OH)₂; "borane reducing agent" means a stable complex of borane and a suitable reagent, such as BH₃ ^»THF, BH₃ ^»S(CH₃)₂ or TBAB; and

"hydride reducing agent" means a metal hydride reagent, such as NaBH , Red-AI, DIBAL-H, L-Selectride, Vitride, LiBH₄, LiAIH₄, LiAI(0tBu)3H, NaCNBH₃, DMAB, zinc borohydride, calcium borohydride, a combination of UBH4 and ZnBr₂, or a combination of NaBH.4 and LiCI.

The term "acyl radical of a naturally occurring amino acid" means a group of the formula -C(0)-R²⁹, wherein R²⁹ is a group of the formula

wherein R³⁰ and R³¹ are the residual portions of said amino acid. For example R³⁰ and R³¹ can be independently selected from H, alkyl or M-substituted alkyl, wherein M is HO-, HS-, CH₃S-, -NH₂, phenyl, p-hydroxyphenyl, imidazolyl or indolyl, such that HO-C(0)-R²⁹ is an amino acid selected from alanine, glycine, valine, leucine, isoleucine, phenylalanine, tryptophan, methionine, serine, threonine, histidine, cysteine or tyrosine.

The following solvents and reagents are referred to herein by the abbreviations indicated: tetrahydrofuran (THF); ethanol (EtOH); methanol (MeOH); acetic acid (HOAc or AcOH); ethyl acetate (EtOAc); N,N-dimethyl- formamide (DMF); trifluoroacetic acid (TFA); trifluoroacetic anhydride (TFAA); 1-hydroxybenzotriazole (HOBT); m-chloroperbenzoic acid (MCPBA); triethylamine (Et₃N); diethyl ether (Et₂0); t-butylamineborane (TBAB); ethyl chloroformate (CIC0₂Et); 1-(3-dimethylamino-propyl)-3- ethyl carbodiimde hydrochloride (DEC); N.N'-carbonyldiimidazole (CDI); 1 ,8-diaza-bicyclo[5.4.0]undec-7-ene (DBU); [0-(7-azabenzotriazol-1 -yl)- 1,1,3,3-tetramethyluronium hexafluorophosphate (HATU); tetrabutyl- ammonium fluoride (TBAF); dicyclohexylcarbodiimide (DCC); N,N- dimethylaminopy dine (DMAP); diisopropylethylamine (Hϋnigs base); [2- (t-butoxy-carbonyloxyimino)-2-phenylacetonitrile] (BOC-ON); 9-fluorenyl- methyl chloroformate (FMOC-CI); sodium bis(2-methoxyethoxy)aluminum hydride (Red-AI); diisobutylaluminum hydride (DIBAL-H); lithium tri-sec- butylborohydride (L-selectride).

Lines drawn into the ring systems indicate that the indicated bond may be attached to any of the substitutable ring atoms.

Certain compounds of the invention may exist in different isomeric forms (e.g., enantiomers, diastereoisomers and geometric isomers). For example, C11 carbon of the tricyclic ring system, (i.e., the point of attachment to the piperazine ring), and the carbon atom of the piperazinyl group to which R² is attached can each independently have the S or R absolute configuration. Various substituent groups, e.g. R¹ , R², can also comprise chiral centers. The invention contemplates all such isomers both in pure form and in admixture, including racemic mixtures. Enol forms are also included, as are the E or Z geometric isomers of compounds which have double bonded substituents, (e.g. where R² is an alkenyl group) .

Certain tricyclic compounds will be acidic in nature, e.g. those compounds which possess a carboxyl or phenolic hydroxyl group. These compounds may form pharmaceutically acceptable salts. Examples of such salts may include sodium, potassium, calcium, aluminum, gold and silver salts. Also contemplated are salts formed with pharmaceutically acceptable amines such as ammonia, alkyl amines, hydroxyalkylamines, N-methylglucamine and the like.

Certain basic tricyclic compounds also form pharmaceutically acceptable salts, e.g., acid addition salts. For example, the pyrido- nitrogen atoms may form salts with strong acid, while compounds having basic substituents such as amino groups also form salts with weaker acids. Examples of suitable acids for salt formation are hydrochloric, sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic, maleic, methanesulfonic and other mineral and carboxylic acids well known to those in the art. The salts are prepared by contacting the free base form with a sufficient amount of the desired acid to produce a salt in the conventional manner. The free base forms may be regenerated by treating the salt with a suitable dilute aqueous base solution such as dilute aqueous NaOH, potassium carbonate, ammonia and sodium bicarbonate. The free base forms differ from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but the acid and base salts are otherwise equivalent to their respective free base forms for purposes of the invention.

All such acid and base salts are intended to be pharmaceutically acceptable salts within the scope of the invention and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the invention.

Certain compounds of the formula (1.0) comprise sulfhydryl groups, (i.e., -CH₂SH), which sulfhydryl groups are capable of reacting to form disulfide bonds resulting in dimeric compounds. An example of such dimers are disulfides of the formula (la). Said sulfhydryl groups can also form disulfides with another thiol, such as glutathione. Disulfides including but not limited to disulfides of formula (la) are within the scope of the invention and are encompassed by the structure of formula (1.0).

Compounds of the formula (1.0) can generally be prepared from an amine of the formula (2.0) as shown in Reaction Scheme 1.

Reaction Scheme 1

For compounds of the formula (1.0) wherein R¹ and the nitrogen atom to which it is attached together comprise an amide, e.g. where R¹ is -C(0)-CH₂-R⁵, the amine (2.0) is reacted with a carboxylic acid of the formula R⁵-CH₂-C(0)-OH in the presence of a coupling agent such as DEC.CDI or DCC. The reaction is typically carried out in a suitable organic solvent such as DMF, THF or CH₂CI₂ at a temperature of -10° to 100°C, preferably at 0° to 50°C, and most preferably at about room temperature. When the coupling agent is DCC or DEC, the reaction is preferably conducted in the presence of HOBT.

Alternatively, the amine (2.0) can be reacted with a compound of the formula R¹-L, wherein R¹ is as defined above and L is a leaving group, such as CI, Br, I , -0-C(0)-R⁴⁰ wherein R⁴⁰ is C Cβ alkyl or phenyl, or a sulfonyl group of the formula -OS0₂-R²°, [wherein R²⁰ is selected from C-ι-C-₆ alkyl, phenyl, CF₃, tolyl and p-bromophenyl], to form a compound of the formula (1.0). The reaction is carried out in the presence of a base, preferably a tertiary amine base, such as Et₃N, DMAP, pyridine or Hϋnigs base.

For preparing compounds of the formula (1.0) wherein R¹ and the nitrogen atom to which it is attached together comprise an amine, e.g. where R¹ is a group of the formula

the amine (2.0) is reacted with an aldehyde of the formula R²¹-CHO, wherein R²¹ is selected such that R¹ corresponds to R²¹-CH₂-, e.g. an aldehyde of the formula

to form an imine of the formula (3.0), wherein R²¹ is as defined above, as shown in Reaction Scheme 2. The imine (3.0) is reduced under suitable reaction conditions to form a compound of the formula (1.0). Preferably the reduction is carried out using a hydride reducing agent, such as NaCNBH₃.

Reaction Scheme 2

When conducting the reactions described above, where R¹ comprises a chemically reactive group, such as amine thiol group, such groups must generally be protected with a suitable protecting group, which can later be removed to complete the synthesis of a compound of formula (1.0). For example, amines can preferably be protected with the BOC protecting group, while thiols can be protected with the trityl (i.e., triphenyimethyl) protecting group. Deprotection, i.e., the removal of these protecting groups is then generally the final step in the synthesis of such compounds of formula (1.0). For preparing compounds of the formula (1.0) wherein R¹ is

-C(0)-NH-R⁵, a compound of the formula (2.0) is reacted with an isocyanate of the formula R⁵-N=C=0, in a suitable solvent such as DMF, MeOH, THF or CH₂CI₂ using methods well known in the art.

Alternatively, an amine (2.0) is reacted with phosgene to form a chloroformate intermediate of the formula (4.0), as shown in Reaction Scheme 3. The chloroformate (4.0) is generally not isolated and is reacted with an amine of the formula R⁵-NH₂, wherein R⁵ is as defined above, to form a compound of the formula (1.0), wherein R¹ is -C(0)-NH-R⁵. Reaction Scheme 3

Compounds of the formula (1.0) wherein R² is -C(0)OR⁶, where R⁶ is other than H, can be prepared from a compound of the formula (1.0) wherein R² is -C0₂H by treating with SOCI₂ or oxalyl chloride, then with an alcohol of the formula R^βOH, wherein R⁶ is as defined above. Similarly, compounds of formula (1.0) wherein R² is -C(0)NR⁶R⁷ can be prepared from a compound of the formula (1.0) wherein R² is -C0₂H via essentially the same method but substituting an amine of the formula R⁶R⁷NH for the alcohol R^H. Alternatively, compounds of formula (1.0) wherein R² is -C(0)NR⁶R⁷ can be prepared by reacting a compound of the formula (1.0) wherein R² is -C0₂H with an amine R⁶R⁷NH in the presence of a coupling agent, such as DCC or DEC. In an analogous manner, compounds of formula (1.0) wherein R² is alkyl substituted by a group of the formula -C(0)OR⁶ or -C(0)NR⁶R⁷ can be prepared from a compound wherein R² is alkyl substituted by -C0₂H via substantially the same procedures as described above.

Compounds of the formula (1.0) wherein R² contains a substituent of formula -S(0)tR⁶, wherein t = 1 or 2, can be prepared by oxidation of an analogous compound of the formula (1.0) wherein R² contains a substituent of formula -S(0)tR⁶ , wherein t = 0, by using a suitable oxidizing agent, such as a peracid, preferably MCPBA.

One skilled in the art will recognize that the above transformations may require protection of sensitive groups in, a compound of the formula (1.0) to be so transformed. For example, for carrying out such transformations on compounds of formula (1.0) where R¹ is a group of the formula

the transformation is typically carried out immediately prior to deprotection of the amine and thiol groups of such R¹ groups.

Amines of the formula (2.0) can be prepared in optically active using appropriate chiral starting materials or alternatively can be prepared using racemic starting compounds to give a mixture of stereoisomeric compounds which can then be separated by resolution or chiral HPLC to give the desired isomer. For example, the amines (2.0) can exist as a mixture of enantiomeric amines, e.g. (2.10) and (2.11), or (2.12) and (2.13), which can be separated by classical resolution techniques using a suitable resolving agent, such as a chiral acid. Chiral acid resolving agents are well known in the art and include such compounds as D- or L- malic acid, D- or L-tartaric acid, di-p-toluoyl-D-tartaric acid, di-p-toluoyl-L- tartaric acid, di-benzoyl-D-tartaric acid and di-benzoyl-L-tartaric acid. Alternatively, the enantiomeric amines (2.11) and (2.10), or (2.12) and (2.13), could be separated using a chiral HPLC column via standard methods.

Amines of the formula (2.0), can be prepared from a piperazine derivative of the formula (5.0), wherein R² is as defined above, and a compound of the formula (6.0), wherein L is a leaving group as defined above and A, B, W and Z are as defined above, via the process shown in Reaction Scheme 4.

Reaction Scheme 4

In the process of Reaction Scheme 4, the piperazine (5.0) is reacted with compound (6.0) in the presence of a base, such as a tertiary amine base, to form a compound of the formula (7.0). Compound (7.0) is then hydrolyzed using a suitable acid, such as HCI or TFA, in a solvent such as dioxane or CH₂CI₂, to form the amine (2.0).

Compounds of the formula (6.0) can be prepared from ketones of the formula (14.0) by the process shown in Reaction Scheme 8.

Reaction Scheme 8

In the process of Reaction Scheme 8, the ketone (14.0) is reduced using a hydride reducing agent, preferably UAIH₄, NaBH₄, UBH₄ or NaCNBH₃, in a suitable solvent, such as THF, Et₂0, or a C₁-C₄ alcohol, at a temperature of -80° to 80°C, preferably at -40° to 60°C, with the temperature and solvent used being selected in accordance with the particular reducing agent employed, to form the alcohol (22.0). In general, boron hydrides, such as NaBH₄ and NaCNBH₃, are used in conjunction with alcohol solvents at a temperature of 0° to 50°C, while the more reactive aluminum hydrides, such as L1AIH₄, are used in solvents such as THF or Et₂0 at a temperature of -40° to 60°C.

The alcohol (22.0) is converted to a compound of formula (6.0). For preparing compounds of formula (6.0) wherein L is halo, the alcohol (22.0) is reacted with a halogenating agent, such as PCI₃, PCI₅, POCI₃, SOCI₂, SOBr₂, 1₂, PBr₃, PBrs, or a combination of Ph3P and either I₂ or Br₂. For preparing compounds of formula (6.0) wherein L is a group of the formula -OCfOz-R⁴⁰ or -OS(0)₂R²², the alcohol (22.0) is reacted with an acid chloride of the formula R⁴⁰C(O)CI or an anhydride of the formula R⁴°C(0)OC(0)R⁴⁰, or a sulfonyl chloride or the formula R ²S(0)₂CI, respectively, in the presence of a base, preferably a tertiary amine base.

Ketones of the formula (14.0) are known or can be prepared by the procedures described in J. Med. Chem.. 4238 (1992), U.S. Patent 5,089,496, and in PCT International Publications WO92/20681 and WO93/02081. For example, intramolecular cyclization of a nitrile of formula (11.0), as defined below, using a strong acid, such as CF₃SO₃H, at a temperature of -15° to 100°C, to form an imine intermediate which is hydrolyzed with water or aqueous acid to form the ketone (14.0).

(11.0) (14.0)

Alternatively, intramolecular Friedel-Crafts acylation of an acid chloride of formula (16.0) may also provide the desired ketone of formula (14.0). The reaction may be carried out under usual Friedel-Crafts conditions in an inert solvent and in the presence of a Lewis acid such as aluminum chloride.

(16.0) (14.0)

Ketones of the formula (14.1), i.e., a compound of the formula (14.0) wherein W is CH, can be prepared by heating a compound of the formula (14.3), i.e., a compound of formula (14.0) wherein W is CH₂, with Se0₂ in acetic acid.

(14.3) (14.1)

Acid chlorides of formula (16.0) can be obtained by hydrolysis of a compound of formula (11.0) to the corresponding carboxylic acid typically by heating with an aqueous acid (e.g., aqueous HCI), followed by conversion of the acid to the acid chloride of (16.0) under standard conditions well known to those skilled in the art (e.g., by treating with SOCI₂ or oxalyl chloride).

Compounds of the formula (11.1), i.e., compounds of the formula (11.0) wherein W is CH₂, are known or can generally be prepared by the process shown in Reaction Scheme 6. According to the process of Reaction Scheme 6 a solution of a compound of the formula (17.0), wherein A is as defined above, in t-butanol is heated in the presence of concentrated H₂Sθ4 to form a t-butylamide of the formula (18.0). The t-butylamide (18.0) is reacted with an alkyllithium reagent, such as n-butyllithium, at -100° to 0°C, preferably at -60° to -20°C, then treated with NaBr and a benzyl halide of formula (19.0), wherein X¹ is CI, Br or I, and B is as defined above, to form a compound of the formula (11.1).

Reaction Scheme 6

(17.0)

Amines of the formula (2.5), wherein X² is Br or I, (i.e., amines of the formula (2.0) wherein A is Br or I), can be prepared via the process shown in Reaction Scheme 7. Reaction Scheme 7

Step A:

Step C:

Step

In Step A of Reaction Scheme 7, a compound of the formula (8.1 ), wherein R²² is C C₆ alkyl, preferably ethyl, and Z, W, B and R² are as defined above, is reacted with a tetraalkylammonium nitrate, such as tetrabutylammonium nitrate, and TFAA in a suitable solvent, such as CH₂CI₂, at -30° to 20°C, preferably at about 0°C, to form a compound of the formula (20.0), wherein R²², B, W, Z and R² are as defined above. In Step B, compound (20.0) is heated with a suitable reducing agent, such as a combination of Fe and CaCI₂, in a polar solvent, such as a C1-C4 alcohol, preferably EtOH, at a temperature of 40° to 100°, preferably at 50° to 80°C, to form a compound of formula (21.0), wherein R²², B, W, Z and R² are as defined above. In Step C, compound (21.0) is converted to the halide (8.2), wherein X² is Br or I, and R²², B, W, Z and R² are as defined above. For forming a compound of formula (8.2) wherein X² is Br, compound (21.0) is treated with Br₂ and HBr at a temperature of -30° to 15°C, preferably at -10° to 10°C, to form the bromide, (i.e., a compound (8.2) wherein X² is Br). For preparing a compound of formula (8.2) wherein X² is I, compound (21.0) is treated with I₂ in a suitable solvent, such as benzene, at a temperature of 30° to 100°C, preferably at 50° to 70°C, to form the iodide, (i.e., a compound (8.2) wherein X² is I).

In Step D, the amine (8.2) is hydrolyzed via substantially the same process as described above for compounds (8.0) and (7.0), to give an amine of the formula (2.5).

Compounds of the formula (5.0) can be prepared via substantially the same methods described in PCT Intemational Publication WO95/00497. Reaction Scheme 12 describes the synthesis of 2-substituted piperazines wherein R² is alkyl, alkenyl, or alkynyl, as well as the synthesis of 2-substituted piperazines wherein R² is alkyl, alkenyl, or alkynyl which are substituted with substituent groups 1), 2), 3), 4) wherein t = 0, or 5), as defined above, with the exception that R⁶ and R⁷ can not be a group that is substituted with -C(0)R¹⁴ or -S0₂R¹⁴.

REACTION SCHEME 12

In Scheme 12, the starting BOC-protected amino acids (32.0) are available commercially or can be made by procedures well known in the art. The amino acids (32.0) can be coupled to N-benzylglycine ethyl ester using a coupling agent such as DCC or DEC in a suitable solvent (e.g., DMF, CHCI3 or CH₂CI₂) to produce a compound of Formula (33.0). Generally, this reaction is conducted at 0° to 35°C, preferably at about 25°C.

The BOC protecting group of compound (33.0) is hydrolyzed via standard methods, such as treatment with an acid, preferably TFA or HCI, in a suitable solvent such as CHCI3 or dioxane at 0° to 50°C, preferably at about 25°C and the deprotected dipeptide is cyclized by treatment with base to produce the compound of formula (34.0).

Compound (34.0) is reduced using a hydride reducing agent, preferably UAIH4 in refluxing Et₂0 or THF to give a piperazine of formula (35.0). The piperazine (35.0) is protected with a BOC group by procedures well known in the art to give the compound of Formula (36.0).

The N-benzyl group of compound (36.0) is removed by catalytic hydrogenation (e.g., using Pd/C and hydrogen gas under pressure of 1 to 100 psi, preferably at about 60 psi, to give the compound of Formula (5.0). Compounds of Formula 5.0, wherein R² represents alkyl, alkenyl or alkynyl substituted with substituent groups 1), 3), 5) or 4) (wherein t = 0), as defined above, wherein R⁶ or R⁷ are substituted with -C(0)R¹⁴ or -S(0)₂R¹⁴ are made according to the process shown in Reaction Scheme 13. Compounds of Formula 5.0, wherein R² represents -C(0)NR⁶R⁷ or -C(0)OR⁶, or wherein R² represents alkyl, alkenyl or alkynyl substituted with a group 4) where t = 1 or 2, 6), 7), 8), 9), 10), 11), 12) or 13), as defined above, are also made according to the process of Reaction Scheme 13. REACTION SCHEME 13

In Reaction Scheme 13, the starting amino acids of formula (37.0), wherein R²⁷ is an alkyl, alkenyl or alkynyl group substituted by an -OH group or a -COOH group (or its corresponding ester) are available commercially or can be made by procedures known in the art. Compound (37.0) is reacted according to the procedures described for the first four steps of Reaction Scheme 12 to produce a compound of Formula (40.0) wherein R²⁸ is a hydroxy substituted alkyl, alkenyl or alkynyl group. Compound (40.0) is then protected with a BOC group and then debenzylated according to the procedures described for steps 5 and 6 of Reaction Scheme 12 to produce a compound of Formula (5.10), i.e., a compound of formula (5.0) wherein R² is a hydroxy substituted alkyl, alkenyl or alkynyl group. A compound of the formula (5.10) where R²⁸ is -CH₂OH can be oxidized to produce the corresponding carboxyl group, i.e., where R² is -COOH. This carboxyl group can then be esterified to produce compounds wherein R² is -C(0)OR⁶, or converted to an amide to produce compounds wherein R² is -C(0)NR⁶R⁷ by procedures well known in the art.

The hydroxy group of R²⁸ of a compound of formula (5.10) can be converted to a leaving group, such as chloro, mesyloxy or tosyloxy, by techniques well known in the art. The leaving group can then be displaced by various nucleophiles, to produce other compounds of formula (5.0) For example, reaction with: an organometallic reagent to produce a compound where R² is substituted by a substituent 1); a thiol to produce a compound where R² is substituted by 4) where t = 0; a sulfenyl reagent to produce a compound where R² is substituted by 4) where t = 1 ; a sulfinyl reagent to produce a compound where R² is substituted by 4) where t=2, or by a substituent 10); an amine to produce a compound where R² is substituted by 5); or an alcohol to produce a compound where R² is substituted by 3). The hydroxy group on R²⁸ of compound (5.10) can also be: acyiated, e.g. with a suitable chloroformate compound, to produce a compound (5.0) wherein R² is substituted by 8) or 9), respectively; or alkylated to produce a compound (5.0) wherein R² with is substituted by 3). When R²⁸ is alkyl having more than one carbon atom, or alkenyl or alkynyl, the hydroxy group can be oxidized, as discussed above, to produce the corresponding carboxyl group (i.e., substituent 13) wherein R⁶ is H. This carboxyl group can be esterified to produce compounds wherein substituent 13) is -C(0)OR⁶ wherein R⁶ is other than H, or converted to amides to produce R² with a 12) substituent, by procedures well known in the art. When the leaving group is displaced by an amine (e.g., HNR⁶R⁷) to produce a substituent 5) as described above, for those substituents wherein at least one of R⁶ or R⁷ is H, the resulting amine substituent 5) can subsequently be converted to R² substituted by 6), 7) or 11) by reacting, with an acyl halide, a carbamyl halide or a sulfonyl halide, respectively, by procedures well known in the art.

Compounds of the formula (5.1), (i.e., racemates of compounds of the formula (5.0) wherein R² is -C(0)NR⁶R⁷ )_> can be prepared from

2-piperazinecarboxylic acid via the process shown in Reaction Scheme 9.

Reaction Scheme 9

In the process of Reaction Scheme 9, 2-piperazine carboxylic acid is treated with BOC-ON in the presence of a hydroxide base, preferably NaOH or KOH, in a suitable solvent, such as a mixture of dioxane and water, then with FMOC-CI under substantially the same conditions to form the differentially protected compound (23.0).

Compound (23.0) is reacted with an amine of the formula R⁶R⁷NH, wherein R⁶ and R⁷ are as defined above, in the presence of HATU in CH₂CI₂. Compound (24.0) is selectively deprotected by treating with piperidine in a suitable solvent, such as DMF, to form a compound of the formula (5.1).

In analogous manner, compounds of formula (5.0) wherein R² is -C(0)OR⁶ can be prepared from compound (23.0) by esterification with an appropriate alcohol R⁶OH using standard methods, followed by deprotection as described for compound (24.0).

Compounds of the formula (5.2), wherein E is -OR⁶ or -NR⁶R⁷, (i.e., racemates of compounds of the formula (5.0) wherein R² is a methylene group substituted by a group of the formula -C(0)OR⁶ or -C(0)NR⁶R⁷), can be prepared via the process shown in Reaction Scheme 10.

Reaction Scheme 10

In the process of Reaction Scheme 10, N.N'-dibenzylethylene- diamine is reacted with methyl 4-bromocrotonate and a tertiary amine base, such as Et3N, in a suitable solvent, such as toluene, to form the N.N'-dibenzylpiperazine derivative (25.0).

Compound (25.0) is hydrogenated over a catalyst, such as Pd/C, to form piperazine derivative (26.0). The 4-amino group of compound (26.0) is then protected as the BOC derivative using BOC-ON to form compound (27.0).

Compound (27.0) is hydrolyzed using a hydroxide base, such as NaOH or KOH, and the free amino group is protected with a suitable amine protecting group, such as an FMOC group to form compound (28.0).

Compound (28.0) is reacted with an amine of the formula R⁶R⁷NH using a coupling agent, such as DEC, in a suitable solvent, such as CH₂CI₂ of DMF, then deprotected using TBAF in DMF to form a compound of the formula (5.2), wherein E is -NR⁶R⁷. Alternatively, compound (28.0) is esterified by reacting with SOCI₂ or oxalyl chloride in the presence of a tertiary amine base to form an acid chloride which is reacted with an alcohol of the formula R⁶OH, then deprotected by treating with TBAF in DMF to form a compound of the formula (5.2) wherein E is -OR⁶.

An alternative method for preparing compounds of the formula (1.0) is shown in Reaction Scheme 14.

Reaction Scheme 14

In Reaction Scheme 14, a compound of the formula (6.0) is reacted with a compound of formula (42.0), wherein R¹ and R² are as defined above for compound (1.0), in a suitable solvent, such as CH3CN or THF, in the presence of a base, such as a tertiary amine base, pentamethylpiperidine or DBU, with pentamethylpiperidine being preferred, to form a compound of formula (1.1).

Compounds of formula (42.0) are prepared as shown in Reaction Scheme 15. Reaction Scheme 15

In Reaction Scheme 15, compound (45.0), wherein R² is as defined above for compound (1.0), the FMOC protecting group is selectively removed, e.g. by reacting with piperidine in a suitable solvent, such as DMF, to form a compound of formula (44.0), which is then converted to a compound of formula (43.0) via substantially the same methods as described above for conversion of compounds of formula (2.0) into compounds of formula (1.0). Compound (43.0) is then deprotected, e.g. by reacting with an acid, such as TFA, in a suitable solvent, such as CH₂CI₂, to form a compound of the formula (42.0).

Compounds of the formula (45.0) can be prepared via substantially the same procedures as described above for preparation of compounds of the formula (24.0) and (28.0), or by similar procedures as those described above for preparing compounds of formula (5.0) by adding additional protection/deprotection steps as necessary.

Compounds of the formula (1.30), wherein p = 0, 1, 2, 3, 4 or 5, (i.e., compound of the formula (1.0) wherein R² is Ci-Cβ alkyl substituted by -OR⁶, wherein R⁶ is H), can be prepared via the process shown in Reaction Scheme 16. Reaction Scheme 16

In the process of Reaction Scheme 16, a compound of formula (46.0) is treated with a suitable borane reducing agent, such as BH3^»THF, in a suitable solvent, such as THF, at a temperature of -40° to 50°C, preferably at 0° to 30°C, to give the alcohol (47.0). The alcohol (47.0) is then protected as a silyl ether, preferably as the TBS ether, by treating with a halotrialkylsilane, preferably chloro-t-butyldimethylsilane, in a suitable solvent, such as CH₂CI₂ or DMF, at 0° to 50°C, preferably at about 25°C, in the presence of a base, such as a tertiaryamine base, preferably EtβN or pyridine, and a catalyst, such as DMAP or imidazole, to give a compound of formula (48.0).

The FMOC protecting group of compound (48.0) can be removed by standard procedures, e.g. by treating with a secondary amine, such as piperidine, in a suitable solvent, such as THF or DMF, at 0° to 50°C, preferably at about 25°C, to give an amine of formula (49.0).

T e amine (49.0) is reacted with a compound of the formula (6.0) in the presence of a hindered base, such as 1 ,2,2,6,6-pentamethylpiperidine of DBU, in a suitable solvent, such as THF or CH3CN, preferably CH3CN, at 0° to 8Q°C, preferably at 25° to 80°C and most preferably at 40° to 60°C, to give a compound of formula (50.0).

The BOC protecting group of compound (50.0) is removed using standard methods, e.g. by treating with an acid, such as TFA or HCI, in a suitable solvent, such as CH₂CI₂ or dioxane, at 0° to 50°C, preferably at about 25°C, to produce a compound of formula (2.20), [i.e., an amine of formula (2.0), wherein R² is -OR⁶ substituted. alkyl, wherein R⁶ is H, and wherein said -OR⁶ group is protected as its TBS ether).

Compound (2.20) is converted to a compound of formula (51.0) via the procedures described above for conversion of an amine (2.0) to a compound of formula (1.0). For example, compound (2.20) can be acyiated with a suitable carboxylic acid in the presence of a coupling agent, such as DCC or DEC, and a base, such as DMAP, in a suitable solvent, such as CH₂CI₂ or DMF, at 0° to 80°C, preferably at about 25°C, to form a compound (51.0), wherein R¹ is -C(0)-CH₂-R⁵ or one of the other acyl groups defined above for R¹. Alternatively, compound (2.20) can be treated with the corresponding acid chloride in the presence of a tertiaryamine base, such as Et3N, to form a compound of formula (51.0), wherein R¹ is -C(0)-CH₂-R⁵ or one of the other acyl groups defined above for R¹.

Compound (51.0) is deprotected using a source of fluoride ion, preferably TBAF, in a suitable solvent, such as THF, at a temperature of 0° to 50°C, preferably at about 25°C, to form a compound of the formula (1.0). Alternatively, compound (51.0) can be treated with aqueous HF in a suitable solvent, such as CH3CN, to produce the compound of formula (1.0).

Compounds of the formula (1.31), wherein p = 0, 1, 2, 3, 4 or 5, (i.e., a compound of the formula (1.0) wherein R² is Ci-Ce alkyl substituted by -OR⁶, wherein R⁶ is -CH₂C(O)NR¹⁰R¹⁵), can be prepared via the process shown in Reaction Scheme 17.

Reaction Scheme 17

In the process of Reaction Scheme 17, compound (52.0) is reduced with a borane reducing agent following substantially the same procedure as described for reduction of compound (46.0) in Scheme 16, to form a compound of formula (53.0).

Compound (53.0) is treated with a strong base such as NaH, in a suitable solvent, such as THF or DMF, preferably THF, at a temperature of -100° to 50°C, preferably at -40° to 10°C, then reacted with an alkyl chloroacetate of the formula CICH₂C(0)OR⁵⁰, wherein R⁵⁰ is C1-C4 alkyl, preferably methyl, to form a compound of the formula (54.0).

Compound (54.0) is hydrolyzed under basic conditions, e.g. by treating with a hydroxide base, such as LiOH, in a suitable solvent, such as a combination of THF, or a C1-C4 alcohol (such as MeOH), and water, at a temperature of 0° to 50°C, preferably at about 25°C, to give a compound of formula (55.0).

Compound (55.0) is reacted with an amine of the formula R¹⁵R¹⁰NH, wherein R¹⁰ and R¹⁵ are as defined above, in the presence of a coupling agent, such as DCC or DEC, preferably DEC, and a catalyst, such as HOBT or DMAP, in a suitable solvent, such as CH₂CI₂ or DMF, at a temperature of 0° to 70°C, preferably at about 25°C, to give a compound of formula (56.0). The N-benzyl protecting group of compound (56.0) is removed by hydrogenolysis using a catalyst, such as Pd/C, preferably 10% Pd/C, in a suitable solvent, such as a C1-C4 alcohol, preferably MeOH or EtOH, at a pressure of 30 psi to 100 psi, preferably at about 50 psi, at a temperature of 0° to 80°C, preferably at 20° to 30°C, to give a compound of the formula (57.0). The hydrogenolysis can more preferably be carried out as described above with a catalytic amount of HOAc present in the mixture during the reaction.

Compound (57.0) is reacted with a compound of the formula (6.0) via substantially the same procedure as described above for the reaction of (6.0) with a compound of formula (49.0) to form a compound of formula (58.0).

The BOC group of compound (58.0) is hydrolyzed as described above for hydrolysis of compound (50.0) to give a compound of the formula (2.21), [i.e., an amine of formula (2.0), wherein R² is -OR⁶ substituted alkyl, wherein R⁶ is -CH₂NR¹⁰R¹⁵]. Compound (2.21 ) is then converted to a compound of the formula (1.31 ) using substantially the same methods as described above for conversion of compound (2.20) to compound (51.0).

Compounds of the formula (1.32), wherein p = 0, 1 , 2, 3, 4 or 5, and f = 2, 3 or 4, (i.e., a compound of the formula (1.0) wherein R² isCi-Cβ alkyl substituted by -OR⁶, wherein R⁶ is C₂-C₄ alkyl substituted by -OH), can be prepared via the process shown in Reaction Scheme 18. Reaction Scheme 18

In the process of Reaction Scheme 18, compound (53.0) is treated with a strong base such as NaH, in a suitable solvent, such as THF or DMF, preferably THF, at a temperature of -100° to 50°C, preferably at -40° to 10°C, then reacted with a protected haloaikanol, e.g. a compound of the formula R⁵¹0-(CH₂) -L, wherein L is halo (preferably Br), f = 2, 3 or 4, and R51 is a suitable hydroxyl protecting group, such as an acyl group (e.g. CH3C(0)-), to form a compound of formula (59.0). The N-benzyl group of compound (59.0) is removed via substantially the same procedure as described for hydrogenolysis of compound (56.0) to form a compound of formula (60.0).

Compound (60.0) is reacted with a compound of the formula (6.0) via substantially the same procedure as described above for the reaction of (6.0) with a compound of formula (49.0) to form a compound of formula (61.0).

The BOC group of compound (61.0) is hydrolyzed as described above for hydrolysis of compound (50.0) to give a compound of the formula (2.22), [i.e., an amine of formula (2.0), wherein R² is alkyl substituted by -OR⁶, wherein R⁶ is alkyl substituted by -OH, where said -OH group is protected, (i.e., by group R⁵¹)]. Compound (2.22) is then converted to a compound of the formula (62.0) using substantially the same methods as described above for conversion of compound (2.20) to compound (51.0).

The R⁵ protecting group of compound (62.0) is then removed using standard conditions appropriate for removal of the protecting group selected. For example, where R⁵¹ is an acyl group, (e.g. CH3C(0)-) compound (62.0) is hydrolyzed using a moderate base, such as K2CO3 or Na₂Cθ3, in a protic solvent, such as a C1-C4 alcohol or water, or a mixture of two such solvents, at a temperature of 0° to 100°C, preferably at about 25°C, to give a compound of formula (1.32). In the above processes, it is sometimes desirable and/or necessary

1 2 to protect certain R , R groups during the reactions. Conventional protecting groups are operable as described in Greene, T.W., "Protective

Groups In Organic Synthesis," John Wiley & Sons, New York, 1981. For example, see Table 1 on page 60 of WO 95/10516 (published April 20, 1995).

Compounds useful in this invention can be prepared by the methods disclosed in WO 95/10516, and by the procedures described in the examples below. The examples should not be construed as limiting the scope of the disclosure. Alternative mechanistic pathways and analogous structures within the scope of the invention may be apparent to those skilled in the art.

PREPARATIVE EXAMPLE 1

8-CHLORO-3-METHYL-5.6-DIHYDRO-11 H-BENZO[5.6|-

CYCLOHEPTAM .2-b1PYRIDIN-11-ONE

By substituting N-(1 ,1 -dimethylethyl)-3,5-dimethyl-2-pyridine carboxamide in step 1 B above and employing basically the same methods as steps B through C of Preparative Example 1 , and Step C of Preparative Example 2, of WO 95/10516, one obtains the title compound. Reaction times are determined by TLC or HPLC. PREPARATIVE EXAMPLE 2

Combine 10 g (60.5 mmol) of ethyl 4-pyridylacetate and 120 mL of dry CH₂CI₂ at -20°C, add 10.45 g (60.5 mmol) of MCPBA and stir at -20°C for 1 hour and then at 25°C for 67 hours. Add an additional 3.48 g (20.2 mmoles) of MCPBA and stir at 25°C for 24 hours. Dilute with CH₂CI₂ and wash with saturated NaHCOβ (aqueous) and then water. Dry over MgSθ₄, concentrate in vacuo to a residue, and chromatograph (silica gel, 2%-

5.5% (10% NH OH in MeOH)/CH₂CI₂)to give 8.12 g of the product compound. Mass Spec: MH+ = 182.15 Step B:

Combine 3.5 g (19.3 mmol) of the product of Step A, 17.5 mL of EtOH and 96.6 mL of 10% NaOH (aqueous) and heat the mixture at 67°C for 2 hours. Add 2 N HCI (aqueous) to adjust to pH = 2.37 and concentrate in vacuo to a residue. Add 200 mL of dry EtOH, filter through celite® and wash the filter cake with dry EtOH (2X50 ml). Concentrate the combined filtrates in vacuo to give2.43 g of the title compound. PREPARAT1VE EXAMPLE 3 σ NHCOOCH₃

Combine 10 g (65.7 mmol) of 3-methoxycarbonylaminopyridine and 150 mL of CH₂CI₂. cool to 0°C and slowly add (dropwise) a solution of 13.61 g (78.84 mmol) of MCPBA in 120 mL of CH₂CI₂ at 0°C over a period of 1 hour. Stir the mixture at 25°C for 5 days, then wash with saturated NaHCθ3 (aqueous), then water and dry over MgSθ4.

Concentrate in vacuo to a residue and chromatograph (silica gel, 2%-5%

(10% NH4OH in MeOH)/CH₂CI₂) to give the product compound. Mass Spec: MH+ = 169

PREPARATIVE EXAMPLE4

Combine 5 g (36.0 mmol) of isonicotinic acid 1-N-oxide and 150 mL of anhydrous DMF, add 5.5 mL (39.6 mmol) of Et3N and stir at 0°C for 0.5 hours. Slowly add (dropwise) 8.5 mL (39.6 mmol) of diphenylphosphoryl azide at 0°C over 10 minutes, stir at 0°C for 1 hour and then at 25°C for 24 hours (as generally described in Pavia, et al., Journal of Medicinal Chemistry. 23, 854-861 (1990). Concentrate in vacuo to a residue and chromatograph (silica gel, 0.5%-1% MeOH/CH₂CI₂) to give 5.9 g of the product compound.

Using nicotinic acid 1-N-oxide and substantially the same procedure as described for Preparative Example 4 the following compound was prepared:

(4A) PREPARATIVE EXAMPLE 5

Hydrogenate 25 g (144 mmol) of 3-pyridylacetic acid hydrochloride for 144 hours using the procedure described in Preparative Example 15, of WO 95/10516, to give 20 g of the product compound. Mass Spec: MH+ = 144. Step B:

React 12 g (83.8 mmol) of the product of Step B for 148 hours using the procedure described in Preparative Example 13, Step B, of WO 95/10516, to give 17.5 g of the product compound. Mass Spec: MH⁺ = 244.25

PREPARATIVE EXAMPLE 6

Combine 25 g (164.4 mmol) of methyl 3-pyridylcarbamate and 163.3 mL of 1N HCI (aqueous), stir until all of the solid dissolves, then hydrogenate over 10% Pd C at 25°C at 55 psi for 220 hours. Filter, wash the solids with water and treat the combined filtrates with 150 mL of BioRad AG1X8 ion exchange resin (OH^*). Filter, wash the resin with water and concentrate the filtrate to a volume of 100 mL. Add 16.43 mL (197.3 mmol) of 37% formalin and hydrogenate over 10% Pd/C at 25°C at 55 psi for 89 hours. Filter, wash the solids with water and concentrate in vacuo to give 24.3 g of the title compound. Mass Spec: MH⁺ = 173.2 PREPARATIVE EXAMPLE 7

Cool 50.0 g (20.5 mmol) of 8-chloro-5,6-dihydro-11 H- benzo[5,6]cyclohepta[1 ,2-b]pyridin-11-one to 0°C, slowly add 75 mL (93.69 mmol) of sulfur monochloride over 20 minutes, then slowly add 25 mL (48.59 mmol) of Br₂ over 15. Heat at 95°C for 20 hour, add 12.5 mL (24.3 mmol) of Br₂ and heat for a another 24 hours. Cool the mixture, and slowly add to a mixture of CH₂CI₂ and 1 N NaOH (aqueous) at 0°C. Wash the organic phase with water, dry over MgS04 and concentrate in vacuo to a residue. Chromatograph the residue (silica gel, 500 mL CH₂CI₂ then 0.2%-5% (10% NH4OH in MeOH)/CH₂CI₂), then chromatograph again (silica gel, 3%-8.5% EtOAc/hexane) to give 8.66 g of the product compound. Mass Spec: MH+ = 322

EXAMPLES 1-5

The title compound from Example 13A of WO 95/00497 is reacted with benzyloxycarbonyl chloride under standard conditions known to one skilled in the art, to give the N-Cbz protected alcohol shown above. After purification in the usual way the latter may be reacted with a variety of reagents shown in Column 1 of Table 1 to give the corresponding N-Cbz protected intermediates where R is as defined in Column 2 of Table 1. After purification in the usual way the latter may be deprotected using mild catalytic hydrogenation procedures known in the art, to give after suitable purification, the final desired intermediates shown in Column 2 of Table 1.

EXAMPLE 6

BOCNH

The title compound from Example 27D of WO 95/00497 is converted by the scheme shown above, using standard procedures known to one skilled in the art, into 1-BOC-2(S)-(4-acetylaminobutyl)- piperazine.

EXAMPLE 7 ^'

STEP A:

Dissolve 5.25 g (25.85 mmol) of 2-piperazine carboxylic acid^«2HCI in 160 mL of 1 :1 dioxane/H₂0, and adjust the pH to 11 with 50% NaOH (aq.). Slowly add (in portions) a solution of 7.21 g (29.28 mmol) of BOC-ON in 40 mL of dioxane while maintaining the pH at 1 1 with 50% NaOH (aq.) during the addition. Stir at room temperature for 5 hours, then cool to 0°C and adjust to pH 9.5 with 50% NaOH(aq.). Slowly add (in portions) a solution of 7.34 g (28.37 mmol) of FMOC-CI in 40 mL of dioxane, maintaining a pH of 9.5 during the addition with 50% NaOH (aq.). Warm the mixture to room temperature and stir for 20 hours. Washed with Et₂0 (3 x 150 mL), adjust to pH = 2-3 with 6N HCI (aq), and extract with toluene (3 x 150 mL). Dry the combine extracts over Na₂S04 and concentrate in vacuo Xo a volume of 150 mL. Chill at -20°C overnight, filter to collect the resulting solids, wash with hexane and dry the solids in vacuo to give 5.4 g of the product compound. STEP B:

Slowly add 2.0 g (9.26 mmol) of 2-nitrobenzylbromide to 37 mL of a 2 M solution of CH3NH₂ in THF, then stir at room temperature for 16 hours. Dilute with 200 mL of EtOAc, wash with water (3 x 60 mL), then dry the organic phase over Na₂S04 and concentrate in vacuo to give 1.53 g of the product compound. STEP C:

Combine 2.74 g (6.05 mmol) of the product of Step A, 4.22 mL of

Hϋnigs base, 2.76 g (7.26 mmol) of HATU, and a solution of 1.00 g (6.05 mmol) of the Product of Step B in 25 mL of CH₂CI₂, and stir at room temperature for 16 hours. Dilute with 75 mL of EtOAc wash successively with 10% HCI (aqueous) (2 x 40 mL), saturated NaHCθ3 (aqueous) (2 x 40 mL) and 40 mL of brine. Dry the organic phase over MgSθ4 concentrate in vacuo to a residue and chromatograph (silica gel, 2% MeOH/CH₂CI₂) to give 2.71 g of the product compound. STEP D;

Combine 1.00 g (1.67 mmol) of the Product of Step C, 8 mL of DMF and 0.18 mL (1.83 mmol) of piperidine, and stir at room temperature for 4 hours. Concentrate in vacuo to a residue and chromatograph (silica gel, 4% MeOH/CH₂CI₂) to give 0.34g of the product compound.

If Steps E, F and G were followed, one could obtain the products indicated. STEP E:

Heat a mixture of the Product of Step D, the chloride product of Preparative Example 40, Step B, Of WO 95/10516, and DBU in THF solution at 60°C for 6 hours. Concentrate in vacuo to a residue and chromatograph (silica gel, 2% to 5% MeOH/CH₂CI₂) to give the Product compound along with a second compound (7i) of the formula:

BOC STEP F;

Treat a solution of the Product of Step E in CH₂CI₂ with TFA at room temperature for 4 hours. Concentrate in vacuo to a residue, add EtOAc and wash successively with saturated K₂C0₃ (aqueous) (twice) and brine. Dry the organic phase over Na₂Sθ4 and concentrate in vacuo to give the Product compound. STEP G;

Combine the product of Step F, 3-pyridylacetic acicHHCI, DMAP,

DCC and CH₂CI₂ and stir at room temperature for 16 hours.

Chromatograph (silica gel, 4% MeOH/CHCI) to give the Product compound.

Using compound (7i) from Step E and following substantially the same procedure as described for Steps F and G, the following compound could be prepared:

Example 7- A

Using substantially the same procedure as described for Example 7, Steps A-G, but substituting the indicated amine for CH₃NH₂ in Step B, and or the indicated acid for 3-pyridylacetic acid in Step G, the following compounds could also be prepared:

STEP A:

Combine 12.05 g (48.5 mmol) of ethyl 1-N-benzyl-2-piperazine- carboxylate in 100 mL of THF with 10.59 g (48.5 mmol) of di-t-butyl- dicarbonate and stir at room temperature for 3 hours. Concentrate in vacuo to give 17.17 g of the product compound. STEP B;

Combine 17.17 g of the product compound fro Step A, 150 mL of MeOH, 7.5 mL of HOAc and 3.4 g of 10% Pd C, and hydrogenate with H₂ (50 psi) for 18 hours at room temperature. Filter through celite®, wash the filter cake with MeOH and concentrate the filtrates in vacuo to a residue. Dissolve the residue in 300 mL of EtOAc and wash successively with saturated Na₂Cθ3 (aqueous) (2 X 150 mL) and 100 mL of brine. Dry over MgSθ4 and concentrate in vacuo to give 1 1.54 g of the product compound. STEP C;

BOC

Combine 0.04 g (0.155 mmol) of the product compound fro Step B, 0.5 mL of CH3CN, and add a solution of 0.069 g (0.201 mmol) of the chloride Product of Preparative Example 40, Step B, of WO 95/10516, and 0.029 g (0.186 mmol) of 1,2,2,6,6-pentamethylpiperidine in 0.5 mL of CH3CN, and stir at 65°C for 20 hours. Cool to 25°C, concentrate in vacuo to a residue and chromatograph (silica gel, 3% MeOH/CH₂CI₂) to give 0.065 g of the product compound. STEP P;

Combine 0.065 g (0.115 mmol) of the product compound from Step C, 1 mL of TFA and 3 mL of CH₂CI₂, and stir at room temperature for 4 hours. Concentrate in vacuo to a residue, add 60 mL of EtOAc and wash successively with 1 M Na₂C03 (aqueous) (2 X 20 mL) and 20 mL of brine. Dry over Na₂S0 and concentrate in vacuo to give 0.038 g of the product compound. STEP E:

Combine 0.015 g (0.0323 mmol) of the product compound from Step D, 0.5 mL of CH₂CI₂, 0.0084 g (0.0485 mmol) of 3-pyridylacetic ackWfCI, 0.006 g (0.0485 mmol) of DMAP and 0.010 g (0.0485 mmol) of DCC, and stir at 25°C for 20 hours. Chromatograph (silica gel, 3% MeOH/CH₂CI₂) to give 0.0061 g of the product compound. ¹H NMR (CDCI₃): 1.19 and 1.32 (t, 3H); 2.25-2.50 (m, 1H); 2.58-3.32 (m, 6H); 3.56-4.31 (m, 7H); 4.42 and 4.88 (br d, 1 H); 5.18 and 5.38 (m, 1 H); 7.08- 7.30 (m, 4H); 7.58 (br s, 2H); 8.32-8.52 (m, 3H).

Using 4-pyridyiacetic acid*HCI and substantially the same procedure as described for Example 8, Step E, the following compound was prepared:

Example 8- A

Analytical data for Example 8-A: 1H NMR (CDCI3): 1.10 and 1.35 (m, 3H); 2.25-2.50 (m, 1 H); 2.58-3.30 (m, 6H); 3.58-4.32 (m, 7H); 4.41 and 4.85 (br d and t, 1 H); 5.16 and 5.36 (m, 1 H); 7.03-7.26 (m, 5H); 7.57 (br s, 1 H); 8.32-8.41 (m, 1 H); 8.41-8.61 (br s, 1H).

EXAMPLE 9

STEP A;

Combine 12 mL (50 mmol) of N.N'-dibenzylethylenediamine, 14 mL (100 mmol) of Et3N and 250 mL toluene at 0°C, add 7 mL (50 mmol) of methyl 4-bromocrotonate (7 mL, 50 mmol), slowly warm to room temperature and stir for 24 hours. Filter, concentrate the filtrate in vacuo to a residue and treat with 10% aqueous HCI (300 mL). Filter again and wash the filtrate EtOAc (2 X 100 mL). Basify the filtrate with K₂Cθ3, extract with EtOAc (3 X 150 mL), wash the combined extracts with brine, dry over MgS04 and concentrate in vacuo to give 13.7g of the product compound. ¹H NMR (CDCI₃) 2.28-2.50 (m, 4H), 2.5-2.75 (m, 4H), 3.1 (bs, 1H), 3.42 (d, 2H), 3.52 (d, 1 H), 3.6 (s, 3H), 3 5 (do 1 H), 7.15-7.35 (m, 1 0 H).

STEP B;

Combine 13.7 g (40 mmol) of the product of Step A, 150 mL of MeOH, 50 mL of 1 N HCI (aqueous) and 3 g of 10% Pd/C and hydrogenate with H₂ (50 psi) for 24 hours. Filter, concentrate the filtrate in vacuo to remove most of the MeOH, and basify with K₂Cθ3 to pH = 9-10. Slowly add 9.8 g (40 mmol) of BOC-ON at 0 °C and stir at 0° for 1 hour. Slowly warm up to room temperature, stir 2 hours, and extract with EtOAc (2 X 200 mL). Treat the combined extracts with 50 mL of 10% HCI (aqueous), wash the aqueous layer with EtOAc, basify with K2CO₃ and extract three times with EtOAc. Wash the combined organic layers with brine, dry over MgSθ4 and concentrate in vacuo to give 7.89 g of the product compound. 1H NMR (CDCI3): 1.4 (s, 9H), 2.31 (dd, 1H), 2.37 (dd, 1 H), 2.55 (b, 1H), 2.69-3.02 (m. 4H), 3.75 (s, 3H), 3.88 (b, 2H). STEP C;

Combine 5.2 g (20 mmol) of the product of Step B, 60 mL of THF, 60 mL of 1 N NaOH (aqueous) and stir at room temperature for 6 hours. Cool to 0°C, add 10% HCI (aqueous) to adjust to pH = 9-10, then add 5.2 g (20 mmol) of FMOC-CI. Stir at room temperature for 6 hours, (adding 1 N NaOH (aqueous) to maintain pH = 9-10) 1 hour, then acidify with 10% HCI to pH = 1. Extract twice, wash the combined organic layers with brine, dry over MgS04 and concentrate in vacuo to give 8.56 g of the product compound. ¹H NMR (CDCI3): 1.4 (s, 9H), 2.5-3.0 (m, 5H), 3.9-4.2 (m, 6H), 4.5 (m, 1H), 7-25 (t, 4H), 7.32(t, 4H), 7.48(d, 4H), 7.75(d, 4H). STEP P;

Combine 460 mg (1 mmol) of the product of Step C, 5 mL of CH₂CI₂, 230 mg (1.2 mmol) of DEC and 130 μL (1.5 mmol) of i-propylamine, and stir at 25°C for 6 hours. Treat with 10 mL of 1 N HCI (aqueous), extract with 30 mL of EtOAc, wash the extract with saturated NaHCθ₃ (aqueous) and dry over Na₂Sθ₄. Concentrate in vacuo to give 454.6 mg of the product compound. 53

STEP E:

Combine a solution of 150 mg (0.3 mmol) of the product of Step D in DMF with 142 mg (0.45 mmol) of TBAF, and stir at 25°C for 0.5 hours. Treat with 5 mL of 1 N HCI (aqueous) and wash with 10 mL of EtOAc. Basify with saturated K₂Cθ₃, extract three times with EtOAc and dry the combined extracts over MgSθ4. Concentrate in vacuo to a residue. Treat the residue with the chloride Product from Preparative Example 40, Step B, of WO 95/10516, via substantially the same procedure as described for Example 8, Step C, to give the product compound. STEP F:

Combine the product of Step E, CH₂CI₂ and TFA, and stir at 25°C for 0.5 hours. Concentrate in vacuo to a residue. Acylate the residue by the same method as Step E of Example 8. Concentrate in vacuo to a residue and chromatograph to give the product compound. ASSAYS

The inhibition of farnesyl protein transferase was assayed by measuring the transfer of [³H]farnesyl from [³H]farnesylpyrophosphate to biotinylated Ras-peptide (biotin-KKSKTKCVIM) using the conditions described below for each 96-well plate to be tested. An assay buffer is prepared consisting of 40 mM Hepes, pH 7.5; 5 mM dithiothreitol; 20 mM magnesium chloride and 0.01 (v/v)% Igepal non- ionic detergent.

A SPA (scintillation proximity assay) bead suspension is prepared consisting of 50 mg of Streptavidin SPA beads (Amersham Life-Science) suspended in 2.5 mL of PBS (phosphate buffered saline). Immediately prior to running the assay a stop solution is prepared consisting of 480 μL of the SPA bead suspension mixed with 6720 μL of a solution consisting of 250 mM EDTA (pH 8.0) and 0.5% Bovine Serium Albumin (Fraction V, 96-99% albumin).

An assay mixture is prepared consisting of 480 μL of assay buffer and 3052.8 μL of water. This mixture is vortexed to homogeneity and 48 μL of the Ras peptide is added. The mixture is vortexed and 15.36 μL of FPP and 3.84 μL of [³H]FPP are added and the mixture vortexed again. 37.5 μL of this assay mixture and 2.5μL of a DMSO solution (at test concentration) of the compound being tested are then added to each well of a Costar polypropylene U-bottom microtiter plate. The plate is sonicated for 15 minutes at 37°C and then shaken for 15 minutes on a plate shaker. 10 μL of the enzyme (recombinant Human farnesyl protein transferase) is added to each well using a Beckman Biomek 2000. The plate is incubated at room temperature for 20 minutes and then quenched with 75 μL of the stop solution. 100 μL of the quenched reaction mixture from each well is then transferred to a Wallac crosstalk-free microtiter plate using a Beckman Biomek 2000. Radioactivity is measured in a Wallac 1450 Microbeta plus liquid scintillation counter. Percent inhibition is calculated relative to an uninhibited control.

FPT IC5₀ (inhibition of farnesyl protein transferase, in vitro enzyme assay), GGPT IC5₀ (inhibition of geranylgeranyl protein transferase, in vitro enzyme assay), COS Cell IC50 (Cell-Based Assay), Cell Mat Assay and in vivo tumor activity could be determined by the methods disclosed in WO 95/10516.

The compounds of Examples 8 and 8-A had an FPT IC₅₀ within the range of 0.01 -10μM.

For preparing pharmaceutical compositions from the compounds described by this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets and suppositories. The powders and tablets may be comprised of from about 5 to about 70 percent active ingredient. Suitable solid carriers are known in the art, e.g. magnesium carbonate, magnesium stearate, talc, sugar, lactose. Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration. For preparing suppositories, a low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted, and the active ingredient is dispersed homogeneously therein as by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool and thereby solidify. Liquid form preparations include solutions, suspensions and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injection.

Liquid form preparations may also include solutions for intranasal administration. Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas.

Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally. The transdermal compositions can take the form of creams, lotions, aerosols and/or emulsions and can be included in a transdermal patch of the matrix or reservoir type as are conventional in the art for this purpose.

Preferably the compound is administered orally.

Preferably, the pharmaceutical preparation is in unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component, e.g., an effective amount to achieve the desired purpose.

The quantity of active compound in a unit dose of preparation may be varied or adjusted from about 0.1 mg to 1000 mg, more preferably from about 1 mg. to 300 mg, according to the particular application. The actual dosage employed may be varied depending upon the requirements of the patient and the severity of the condition being treated. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day if desired. The amount and frequency of administration of the compounds of the invention and the pharmaceutically acceptable salts thereof will be regulated according to the judgment of the attending clinician considering such factors as age, condition and size of the patient as well as severity of the symptoms being treated. A typical recommended dosage regimen is oral administration of from 10 mg to 2000 mg/day preferably 10 to 1000 mg/day, in two to four divided doses to block tumor growth. The compounds are non-toxic when administered within this dosage range.

The following are examples of pharmaceutical dosage forms which contain a compound of the invention. The scope of the invention in its pharmaceutical composition aspect is not to be limited by the examples provided.

Pharmaceutical Dosage Form Examples

EXAMPLE A Tablets

No. Ingredients mg/tablet mg/tablet

1. Active compound 100 500

2. Lactose USP 122 1 13

3. Corn Starch, Food Grade, 30 40 as a 10% paste in Purified Water

4. Corn Starch, Food Grade 45 40

5. Magnesium Stearate 3 7

Total 300 700 Method of Manufacture

Mix Item Nos. 1 and 2 in a suitable mixer for 10-15 minutes. Granulate the mixture with Item No. 3. Mill the damp granules through a coarse screen (e.g., 1/4", 0.63 cm) if necessary. Dry the damp granules. Screen the dried granules if necessary and mix with Item No. 4 and mix for 10-15 minutes. Add Item No. 5 and mix for 1-3 minutes. Compress the mixture to appropriate size and weigh on a suitable tablet machine.

EXAMPLE B Capsules

No. Ingredient mg/capsule mg/capsule

1. Active compound 100 500

2. Lactose USP 106 123

3. Corn Starch, Food Grade 40 70

4. Magnesium Stearate NF 7 mmj.

Total 253 700 Method of Manufacture

Mix Item Nos. 1, 2 and 3 in a suitable blender for 10-15 minutes. Add Item No. 4 and mix for 1-3 minutes. Fill the mixture into suitable two- piece hard gelatin capsules on a suitable encapsulating machine.

While the present invention has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications and variations thereof will be apparent to those of ordinary skill in the art. All such alternatives, modifications and variations are intended to fall within the spirit and scope of the present invention.

Claims

WHAT IS CLAIMED IS:

A compound of the formula

wherein:

A and B are independently selected from H, halo or C -Cβ alkyl;

Z is N or CH;

W is CH, CH₂, O or S, wherein the dotted line to W represents a double bond which is present when W is CH;

R¹ is selected from the group consisting of:

wherein W, A and B are as defined above; or R¹ is a group D, wherein D is -C(0)-(CH₂)_s-R⁵, -C(0)0-(CH₂)_m-R⁵ or -C(0)NH-(CH₂)_m-R⁵, wherein R⁵ is aryl, heteroaryl, heterocycloalkyi, or a group of the formula

wherein g = 1 or 2, and R¹¹ represents H, Ci-Cβ alkyl, haloalkyi or -C(O)-

R⁹ wherein R⁹ is C Cβ alkyl, Ci-Cβ alkoxy or -NH(R^10A) wherein R10A j_s

H or alkyl, or the group -C(0)-R⁹ represents an acyl radical of a naturally occurring amino acid; or

R¹ is a group of the formula:

wherein:

(a) T is selected from: o o II O

-C-N— II

H , ^c~ °~~ , -S0₂-, or a single bond;

(b) x is 0, 1 , 2, 3, 4, 5 or 6;

(c) each R^a and each R^b is independently selected from H, aryl, alkyl, alkoxy, aralkyi, amino, alkylamino, heterocyloalkyl, -COOR⁶⁰, -NHfCfOJkR⁶⁰ (wherein z is 0 or 1), or -(CH)_wS(0)_tR⁶⁰ (wherein w is 0, 1 , 2 or 3, and t is 0, 1 or 2); or R^a and R^b taken together can represent cycloalkyl, =N-0-alkyl, =0 or heterocycloalkyi; with the proviso that for the same carbon, R^a is not selected from alkoxy, amino, alkylamino or -NH{C(0)}_zR⁶⁰ when R^b is selected from alkoxy, amino, alkylamino or -NH{C(0)}_zR⁶⁰; and with the proviso that when T is a single bond, for the first carbon containing R^a and R^b, R^a and R^b are not selected from alkoxy, alkylamino, amino or -NHR⁶⁰; and

(d) R⁹² can represent H, alkyl, aryl, aryloxy, arylthio, aralkoxy, aralkyi, heteroaryl or heterocycloalkyi;

R⁶⁰ represents H, alkyl, aryl or aralkyi; R⁴ is H or Ci-Ce alkyl; R² is selected from: -C(0)OR⁶, -CfOJNRW, Cι-C₈ alkyl, C₂-C₈ alkenyl, C₂-Cβ alkynyl, substituted (Cι-C8)alkyl, substituted (C₂-Cβ)alkenyl, substituted (C₂-Cβ)alkynyl, wherein said substituted groups have one or more substituents selected from: 1) aryl, heteroaryl, heterocycloalkyi, B-substituted aryl,

B-substituted heteroaryl or B-substituted heterocycloalkyi, wherein B is selected from C₁-C4 alkyl, phenyl, -(CH₂)_nOR⁶, -(CH₂)_nNR⁶R⁷ and halo;

2) C3-C6 cycloalkyl;

3) -OR⁶; 4) -S(0)_tR⁶;

5) -NR⁶R⁷;

6) -N(R⁶)-C(0)R⁷;

7) -N(R⁶)-C(0)NR⁷R1²;

8) -0-C(0)NR⁶R⁷; 9) -0-C(0)OR⁶;

10) -S0₂NR⁶R⁷;

11) -N(R⁶)-S0₂-R⁷;

12) -C(0)NR⁶R⁷;

13) -C(0)OR⁶; and provided that: where R¹ is D, R² is not H; where R¹ is D and R² is Ci-Cβ alkyl, the substituents on said alkyl group are not substituents 4), 5), 9) or 13); and where R¹ is D, and R² is Ci-Cβ alkyl substituted by the group -OR⁶, R⁶ is not H, alkyl, aryl, substituted aryl, aryl-substituted alkyl or nitro- phenylsubstituted alkyl; R⁶, R⁷ and R¹² are independently selected from H, C₁-C₄ alkyl,

(C₃-C6)cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyi, substituted (Cι-C4)alkyl, substituted (C₃-Ce)cycloalkyl, substituted aryl, substituted arylalkyl, substituted heteroaryl, substituted heteroarylalky or substituted heterocycloalkyi, wherein said substituted groups have one or more substituents selected from: C₁-C₄ alkoxy, aralkyi, heteroarylalkyl, -N0₂, (C₃-Cιo)alkoxyalkoxy, (C₃-C6)cycloalkyl, aryl, -CN, nitro-phenyl, methylenedioxyphenyl, heteroaryl, heterocycloalkyi, halo, -OH, -COOH, -C(0)R¹⁴, -C(0)OR¹⁴, -C(0)NR⁶R⁷, -N(R⁶)C(0)R¹⁴, -S(0)_tR¹⁴ or -NR¹⁰R¹⁵; provided that R⁶, R⁷ and R ² are not -CH₂OH or -CH₂NR¹⁰R¹⁵ when said R⁶, R⁷ or R¹² is directly bonded to a heteroatom, and further provided that R⁶ is not H for groups 4) and 9), and R⁷ is not H for group 6); optionally, when R⁶ and R⁷ are bound to the same nitrogen, R⁶ and R⁷ together with the nitrogen to which they are bound, form a 5 to 7 membered heterocycloalkyi ring which optionally contains O, NR⁶, or S(0)t wherein t is 0, 1 or 2; optionally, when R⁷ and R¹² are bound to the same nitrogen, R⁷ and R¹² together with the nitrogen to which they are bound, form a 5 to 7 membered heterocycloalkyi ring which optionally contains O, NR⁶, or S(0)t wherein t is 0, 1 or 2;

R⁸, R¹⁰ and R¹⁵ are independently H, C1-C4 alkyl or arylalkyl; R¹⁴ is C1-C4 alkyl, aryl or arylalkyl; m = 0, 1, 2 or 3; n = 0, 1 , 2, 3 or 4; s = 1 , 2 or 3; and t = 0, 1 or 2; or pharmaceutically acceptable salts thereof.

2. A compound of claim 1 wherein s = 1 , m = 0 and R⁵ is selected from phenyl, B-substituted phenyl, pyridyl, pyridyl N-oxide, or a piperidinyl group of the formula

3. A compound of claim 2 wherein R² is -COOR⁶, A is Br, B is CI, W is CH₂ and R¹ is -C(0)-CH₂-pyridyl.

4. A compound of claim 3 wherein R⁶ is ethyl and R¹ is

5. A method for inhibiting the abnormal growth of cells comprising administering an effective amount of a compound of claim 1.

6. The method of Claim 5 wherein the cells inhibited are tumor cells expressing an activated ras oncogene.

7. The method of Claim 5 wherein the cells inhibited are pancreatic tumor cells, lung cancer cells, myeloid leukemia tumor cells, thyroid follicular tumor cells, myelodysplastic tumor cells, epidermal carcinoma tumor cells, bladder carcinoma tumor cells or colon tumors cells.

8. The method of Claim 5 wherein the inhibition of the abnormal growth of cells occurs by the inhibition of farnesyl protein transferase.

9. The method of Claim 5 wherein the inhibition is of tumor cells wherein the Ras protein is activated as a result of oncogenic mutation in genes other than the Ras gene.

10. A pharmaceutical composition for inhibiting the abnormal growth of cells comprising an effective amount of compound of Claim 1 in combination with a pharmaceutically acceptable carrier.

11. The use of a compound of Claim 1 for inhibiting the abnormal growth of cells .

12. The use of a compound of Claim 1 for the manufacture of a medicament for inhibiting the abnormal growth of cells.