MXPA99012061A - Benzo(5,6)cyclohepta(1,2b)pyridine derivatives useful for inhibition of farnesyl protein transferase - Google Patents

Abstract

Novel compounds of formula (1.0) or a pharmaceutically acceptable salt or solvate thereof, wherein:a represents N or NO-;R1 and R3 are the same or different and each represents halo;R2 and R4 are the same or different and each is selected from H and halo, provided that at least one of R2 and R4 is H;T is a substituent selected from SO2 R or (A);Z is O or S;n is zero or an integer from 1 to 6;R is alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, or N(R5)2;R5 is H, alkyl, aryl, heteroaryl or cycloalkyl. Also disclosed are methods of inhibiting farnesyl protein transferase and methods for treating tumor cells.

Description

BENZO DERIVATIVES (5, 6) CICLOHEPT (1, 2B) USEFUL PIRIDINE FOR INHIBITION OF FARNESIL TRANSFER PROTEIN , BACKGROUND EJ. WO 95/10516, published April 20, 1995 discloses tricyclic compounds useful for inhibiting farnesyl protein transferase. In view of the current interest in farnesyl protein transferase inhibitors, a welcome contribution to the art would be useful compounds for the inhibition of farnesyl protein transferase. Such contribution is provided by this invention.

BRIEF DESCRIPTION OF THE INVENTION This "invention provides compounds useful for the inhibition of farnesyl protein transferase (FPT)." The compounds of this invention are represented by the formula: or a pharmaceutically acceptable salt or solvate thereof, wherein: a represents N or NO ", - R1 and R3 are the same or different and each represents halo, - R2 and R4 are the same or different and each is selected from H and halo, with the proviso that at least one of R2 and R4 is H; T is a substituent selected from S02R or Z is O or S; n is "zero or an integer from 1 to 6, R is alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl or N (R5) 2; Rs is H, alkyl, aryl, heteroaryl or cycloalkyl. this invention.- (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 transformant Ras engineered to be a geranylgeranyl acceptor, - (iii) they block the intracellular processing of Ras which is a farnesyl acceptor, but not of Ras engineered to be a geranylgeranyl acceptor; iv) block the abnormal growth of cells in culture induced by transforming Ras The compounds of this invention inhibit famesyl protein transferase and farnesylation of the Ras oncogene protein. The invention further provides a method for inhibiting farnesyl protein transferase (eg, ras farnesyl protein transferase) in mammals, especially humans, by administering an effective amount of the compounds of formula 1.0. The administration of the compounds of this invention to patients to inhibit farnesyl protein transferase is useful in the treatment of cancers described below.

This invention provides a method for inhibiting or treating the abnormal growth of cells, including transformed cells, by administering an effective amount of a compound of this invention. Abnormal cell growth refers to the growth of cells independent of normal regulatory mechanisms (e.g., loss of contact inhibition). This includes the abnormal growth of: (1) tumor cells (tumors) that express an activated Ras oncogene; (2) tumor cells in which the Ras protein is activated as a result of the oncogenic mutation in another gene; and (3) benign and malignant cells of other proliferative diseases in which aberrant Ras activation occurs. This invention also provides a method for inhibiting or treating 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 or treating the growth of tumors expressing a Ras oncogene activated by the administration of an effective amount of the compounds of formula 1.0. Examples of tumors which may be inhibited or treated, include, but are not limited to, lung cancer (e.g., lung adenocarcinoma), pancreatic cancers (e.g., pancreatic carcinoma such as, e.g., exocrine pancreatic carcinoma), colon cancers (eg, colorectal carcinomas such as, for example, colon adenocarcinoma and colon adenoma), myeloid leukemias (e.g., acute myelogenous leukemia (AML)), follicular thyroid cancer, myelodysplastic syndrome (MDS), carcinoma of the vejiga, epidermal carcinoma, breast cancer and prostate cancer. It is considered that this invention also provides a method for inhibiting or treating both benign and malignant proliferative diseases, wherein the Ras proteins have been aberrantly activated as a result of oncogenic mutation in other genes - that is, the Ras gene itself is not has been activated by mutation to an oncogenic form - wherein such inhibition or treatment is carried out by the administration of an effective amount of a compound of formula 1.0 to a mammal (eg a human) in need of such treatment. For example, benign proliferative disorder of neurofibromatosis or tumors in which Ras is activated due to mutation or overexpression of tyrosine kinase oncogenes (eg, neu, src, abl, lc and fyn), can be inhibited or treated by tricyclic compounds described in the present. The compounds of formula IO useful in the methods of this invention inhibit or treat abnormal cell growth. Without wishing to be bound by any theory, it is considered that these compounds can function through the inhibition of G protein function, such as ras p21, by blocking the isoprenylation of G protein, thus rendering them useful in the treatment of proliferative diseases such as tumor growth and cancer. Without wishing to be bound by any theory, these compounds are considered to inhibit ras farnesyl protein transferase and therefore show antiproliferative activity against ras transformed cells.

DETAILED DESCRIPTION OF THE INVENTION As used herein, the following terms are used as defined below, unless otherwise indicated: MH + - represents the molecular ion plus hydrogen of the molecule in the mass spectrum; Et (or ET) - represents ethyl (C2HS), - alkyl - represents straight and branched carbon chains containing from one to twenty carbon atoms, preferably one to six carbon atoms, - halo - represents fluoro, chloro, bromo and iodo; cycloalkyl - represents branched or unbranched saturated carbocyclic rings of 3 to 20 carbon atoms, preferably 3 to 7 carbon atoms, - heterocycloalkyl - represents a saturated, branched or unbranched carbccyclic ring containing from 3 to 15 carbon atoms, preferably from 4 to 6 carbon atoms, carbocyclic ring which is interrupted by 1 to 3 hetero groups which are selected from -0-, -S-, or -NR9- [where R9 can be, for example, -C ( O) N (R10), -CH2C (O) N (R10) 2, -S02Rx -SO ^ R1 -C (0) Rn, -CiOJ-O-R11, alkyl, aryl, aralkyl, cycloalkyl, heterocycloalkyl or heteroaryl; each R10 independently represents H, alkyl, aryl or aralkyl (for example benzyl); and R11 is alkyl, aryl, aralkyl, heteroaryl or heterocycloalkyl) - suitable heterocycloalkyl groups include 2- or 3-tetrahydrofuranyl, 2- or 3-tetrahydrothienyl, 2-, 3- or 4-piperidinyl, 2- or 3-pyrrolidinyl, 2- or 3-piperizinyl, 2- or 4-dioxanyl, etc., - with preferred heterocycloalkyl groups 2-, 3- or 4-piperidinyl is substituted with R 10 in the piperidinyl nitrogen), - aryl (which includes the aryl portion) of aryloxy and aralkyl) - represents a carbocyclic group containing from 6 to 15 carbon atoms and having at least one aromatic ring (for example aryl is a phenyl ring) with all available substitutable carbon atoms of the carbocyclic group considered as possible binding sites, the carbocyclic group is optionally substituted (for example by 1 to 3) with one or more of halo, alkyl, hydroxy, alkoxy, phenoxy, CF3, amino, alkylamino, dialkylamino, -COOR11 or -N02 (wherein R11 is H, alkyl, aryl, heteroaryl or cycloal chyl), - and heteroaryl - represent cyclic groups, optionally substituted with R3 and R4, having at least one heteroatom selected from O, S or N, the heteroatom disrupts a carbocyclic ring structure and has a sufficient number of electrons pi delocalised to provide an aromatic character, wherein the aromatic heterocyclic groups preferably contain from 2 to 14 carbon atoms, for example triazolyl, 2-, 3-, or 4-pyridyl or pyridyl N-oxide (optionally substituted with R11 as defined above) where pyridyl N-oxide can be represented as: The following solvents and reagents are referred to herein by the indicated abbreviations: ethanol (EtOH); methanol (MeOH); acetic acid (HOAc or AcOH); ethyl acetate (EtOAc); N, N-dimethylfortnamide (DMF); trifluoroacetic acid (TFA), - trifluoroacetic anhydride (TFAA); 1-hydroxybenzotriazole (HOBT); l- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (DEC); diisobutylaluminum hydride (DIBAL) and 4-methylmorpholine (NMM). The positions in the tricyclic ring system are: Those skilled in the art will also appreciate that the stereochemistry S and R for the C-ll position of the tricyclic ring is as follows: Preferred halo atoms for R1, R2, R3 and R4 in formula 1.0 are selected from: Br, Cl or I, with Br and Cl being preferred. Compounds of formula 1.0 include compounds of the formula: wherein R1 and R3 are the same or different halo and a and T are as defined above. Preferably, for these dihalo compounds, R1 and R3 are independently selected from Br or Cl, and more preferably R1 is Br and R3 is Cl. The compounds of formula 1.0 include compounds of formula 1.1 and 1.2: . 2 : - li where R1, R3 and R4 in formula 1. 1 are halo, and R1, R2 and R3 in formula 1. 2 are halo. The compounds of formula 1 are preferred. 1 . Preferably, in formula 1.1, R1 is Br, R3 is Cl and R4 is halo. More Preferably, in formula 1.1, R1 is Br, R3 is Cl and R4 is Br. Preferably, in formula 1.2, R1 is Br, R2 is halo and R3 is Cl. More preferably, in formula 1.1, R1 is Br, R2 is Br and R3 is Cl. In addition, preferably for the compounds of this invention, substituent a on ring I represents N. T preferably is a group -S02 methyl or a group wherein R is a 3-pyridinyl N-oxide, 4-pyridinyl N-oxide, 4-piperidinyl, 3-piperidinyl or 3-pyrrolidinyl group, wherein the 4-piperidinyl, 3-piperidinyl or 3-pyrrolidinyl groups may be substituted in the piperidinyl or pyrrolidinyl nitrogen with a group R9 which may be, for example, -C (0) N (R10) 2, -CH2C (O) N (R10) 2, -S02R10, -SO2N (R10) 2 , -C (0) RX1; CÍOÍOR11, alkyl, aryl, aralkyl, cycloalkyl, heterocycloalkyl or heteroaryl; each R 10 independently represents H, alkyl, aryl or aralkyl (for example benzyl), and R 11 is alkyl, aryl, aralkyl, heteroaryl or heterocycloalkyl. Those skilled in the art will appreciate that the compounds of formula 1.0 include compounds of formulas 1.3 and 1.4: the compounds of 1.3 being preferred for compounds of formula 1.1 and the compounds of formula 1.4 being preferred for the compounds of formula 1.2. Therefore, the compounds of the invention include compounds of the formulas: ) Certain compounds of the invention may exist in different isomeric forms (e.g. enantiomers and diastereomers). The invention contemplates all types of isomers in both pure form and in mixture, including racemic mixtures. Enol forms are also included. Certain compounds of formula 1.0 will be of acidic nature, for example those compounds which possess a carboxyl or phenolic hydroxyl group.These compounds can form pharmaceutically acceptable salts Examples of such salts may include sodium, potassium, calcium, aluminum salts, Gold and silver The salts formed with pharmaceutically acceptable amines such as ammonia, alkylamines, hydroxyalkylamines, N-methylglucamine and the like are also contemplated Certain basic compounds of formula 1.0 also form pharmaceutically acceptable salts, for example acid addition salts. , pyridium nitrogen atoms can form salts with a 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 skilled 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 can 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 the respective salt forms to some extent, in some physical properties such as solubility in polar solvents, but the acid and base salts are otherwise equivalent to their respective free base forms for the purposes of the invention. It is intended that all acid and base salts be pharmaceutically acceptable salts within the scope of the invention and that all acid and base salts be considered equivalent to free forms for the corresponding compounds, for the purposes of the invention. Intermediates useful for the preparation of the compounds of the invention can be prepared according to the methods described in WO 95/10516 published April 20, 1995, in WO 96/30363 published October 3, 1996, in the patent of the United States number 5,151,423 and by the methods described in the following. The compounds of the invention can be prepared according to the reaction: .2) In the reaction, the carboxylic acid (14.0) is coupled to the tricyclic amine (13.0) using the amide bond formation conditions well known to those skilled in the art. The substituents are as defined for formula 1.0. For example, carbodiimide coupling methods (eg DEC) can be used. For example, the carboxylic acid (14.0) can be reacted with a tricyclic amine (13.0) using DEC / HOBT / NMM in DMF at about 25 ° C for a sufficient period of time, for example about 18 hours, to produce a compound of formula 1.0. When T is S02R, X is halo, preferably chlorine.

The tricyclic piperadine compound is dissolved in an appropriate solvent such as DMF or THF. A base such as triethylamine is added and the appropriate alkylsulfonyl chloride prepared by methods known in the art is added to the reaction mixture at 0 ° C at room temperature with stirring. After 1-24 hours, the reaction mixture is added to water and the product is extracted with a suitable solvent such as ethyl acetate. The crude reaction product is then subjected to chromatography on a column of silica gel. Alkylaminosulfonamido derivatives can similarly be prepared: wherein the groups R5 may be the same or different and each is as defined above. In this reaction, the tricyclic piperadine compound is dissolved in an appropriate solvent such as DMF or THF. A base such as triethylamine is added, and the appropriate alkylaminosulfonyl chloride, prepared by methods known in the art, is added to the reaction mixture, from 0 ° C to room temperature, with stirring. After 1-24 hours, the reaction mixture is added to water and the product is extracted with a suitable solvent such as ethyl acetate. The crude reaction product can then be chromatographed on a column of silica gel. The carboxylic acids (14.0) and the sulfonates (14.2) are generally well known in the art or can be prepared by methods well known in the literature. The compounds of formula 13.0 can be prepared from compounds of formula 13.0a: wherein R6 is H, alkyl, carboalkoxy or any other group that can be converted to a T group. The compounds of formula 13.0a are prepared by methods known in the art, for example, by methods described in WO 95/10516 , in WO 96/30363 published October 3, 1996, in U.S. Patent No. 5,151,423 and those described below. The double bond in the compounds of formula 13.0a can be removed by oxidation, for example, by the method in preparative example 3 below, to provide the ketones of formula 15.0 below: . Compounds of formula 13. O where the C-3 position of the pyridine ring in the tricyclic structure is substituted by bromine (ie, R1 is Br) can also be prepared by a process comprising the following steps: (a) react an amide of the formula wherein RSa is hydrogen and R6a is aryl alkyl or heteroaryl; R5a is C_-C3 alkyl / aryl or heteroaryl; and R a is hydrogen; R5a and R6a are independently selected from the group consisting of C * alkyl and aryl; or RSa and R6a, together with the nitrogen to which they are attached, form a ring comprising 4 to 6 carbon atoms or comprising 3 to 5 carbon atoms and a hetero portion which is selected from the group consisting of -O- and -NR9a-, wherein R9a is H, C-Cg alkyl or phenyl; with a compound of the formula: R wherein R2, R3 and R4 are as defined above, and R7a is Cl or Br, in the presence of a strong base, to obtain a compound of the formula (b) reacting a compound of step (a) with P0C13 to obtain a cyano compound of the formula (c) reacting the cyano compound with a piperidine derivative of the formula: wherein L is halo which is selected from the group consisting of Cl and Br, to obtain a ketone of the following formula: (d) (i) cyclizing the ketone under acidic conditions (for example aluminum chloride, triflic acid or sulfuric acid) to obtain a compound of formula 13.0a wherein R6 is methyl, which can be separated to provide the compound of formula 15.0.

The methods for preparing the compounds of formula 13.0a described in WO 95/10516, in WO 96/30363 published October 3, 1996 and in U.S. Patent No. 5,151,423 and which are described below use an intermediate of tricyclic ketone. Such intermediaries of the formula . 0 wherein R1, R2, R3 and R4 are as defined above, they can be prepared by the following process, which comprises: (a) reacting a compound of the formula (i) with an amine of the formula NHR5aR6a, wherein R5a and RSa are as defined in the above process; in the presence of a palladium and carbon monoxide catalyst to obtain an amide of the formula: (ii) with an alcohol of the formula R10aOH, wherein R10a is lower alkyl of C.-Cg or C3-C6 cycloalkyl, in the presence of a palladium and carbon monoxide catalyst to obtain the ester of the formula followed by reaction of the ester with an amine of formula NHR5aRβa to obtain the amide; (b) reacting the amide with a benzyl compound substituted with iodine of the formula: wherein R2, R3, R4 and R7a are as defined above, in the presence of a strong base, to obtain a compound of the formula: (c) cyclizing a compound of step (b) with a reagent of the formula R8aMgL, wherein R8a is C-C8 alkyl, aryl or heteroaryl and L is Br or Cl, with the proviso that prior to cyclization , compounds in which RSa or R6a is hydrogen react with a suitable N-protecting group. The compounds of formula 1.0, wherein the substituent a is NO (ring I) can be made from compounds of formula 13.0a using procedures well known to those skilled in the art. For example, the compound of formula 13.0a can be reacted with m-chloroperoxybenzoic acid in a suitable organic solvent, for example dichloromethane (usually anhydrous) or methylene chloride, at a suitable temperature to produce a compound of formula 13.0b. which can be subsequently separated to provide a compound of formula 15.0 above. Generally, the organic solvent solution of formula 13.0a is cooled to about 0 ° C before the m-chloroperoxybenzoic acid is added. The reaction is then allowed to warm to room temperature during the reaction period. The desired product can be recovered by a standard separation medium. For example, the reaction mixture can be washed with an aqueous solution of a suitable base, for example saturated sodium bicarbonate or NaOH (for example 1N NaOH) and then dried over anhydrous magnesium sulfate. The solution containing the product can be concentrated in vacuo. The product can be purified by standard means, for example, by chromatography using silica gel (for example, flash column chromatography). Alternatively, the compounds of formula 1.0, wherein the substituent a is NO, can be made from compounds of formula 1.0, wherein substituent a is N by the process of oxidation of m-chloroperoxybenzoic acid described above. Those skilled in the art will appreciate that it is preferable to avoid an excess of m-chloroperoxybenzoic acid when the oxidation reaction is carried out on compounds of formula 13.0a. In these reactions, an excess of m-chloroperoxybenzoic acid can cause oxidation of the double bond in C-ll. Compounds of formula 1.0, wherein Z is S can be prepared from compounds of formula 1.0 wherein Z is 0 by treatment with a suitable sulfur transfer reagent such as a sson reagent. The compounds of the invention having asymmetric carbons (for example compounds of the invention in which X is CH or N having an asymmetric carbon at the C-ll position of the tricyclic ring) can be separated into enantiomers by techniques known in the art. , for example by resolution of chiral salts or by chiral HPLC.

The compounds useful in this invention are exemplified by the following examples, which should not be considered as limiting the scope of the invention.

Preparatory Example 1 3, 10 -dibromo-8-chloro-6,1-dihydro-11-on-5H-benzo [5,6] -cycloheptyl- [1,2-b] pyridine (2 g, 4.98 mmol) is dissolved in water. ml of dry tetrahydrofuran under an atmosphere of dry nitrogen. 5 ml of a 1.5 molar solution of N-methyl-piperidin-4-magnesium chloride is added and the reaction is stirred for 18 hours. The reaction mixture is washed with saturated ammonium chloride, dried over magnesium sulfate, filtered and evaporated to a brown oil which is chromatographed on silica gel using 2.5% methanol / methylene chloride as the eluent for get 2.11 g, from 85% 3, 10-dibromo-8-chloro-6,11-dihydro-11- (1-methyl-4-piperidinyl) -5H-benzo [5,6] cycloheptyl [1,2-b] pyridin-11-ol. EMBAR (M + H) = 501.

Preparation e-j 2 Stage A: .86 g (55.9 mmoles) of 4 - (8 - c 1) or ro - 3 - b - or - 5,6 - dihydro - 11 H - benzo [5,6] cyclohepta [1, 2 - b] ethyl ester are combined. ] pyridin-11-ylidene-1-piperidine-1-carboxylic acid and 250 ml of concentrated H2SO4 at -5 ° C, and then 4.8 g (56.4 mmoles) of NaN03 are added, and the mixture is stirred for 2 hours. The mixture is poured into 600 g of ice and basified with concentrated NH40H (aqueous). The mixture is filtered, washed with 300 ml of water and then extracted with 500 ml of CH2C12. The extract is washed with 200 ml of water, dried over MgSO 4, and then filtered and concentrated in vacuo to a residue. Chromatograph the residue (silica gel, 10% EtOAc / CH2C12) to provide 24.4 g (86% yield) of the product, m.p. = 165-167 ° C, Mass spectrum: MH + = 506 (Cl). Elementary Analysis: Calculated - C, 52.13; H, 4.17; N, 8.29; found - C, 52.18; H, 4.51; N, 8.16.

Stage B: _ g (40.5 mmol) of the product from step A and 200 ml of concentrated H2SO4 are combined at 20 ° C, then the mixture is cooled to 0 ° C. 7.12 g (24.89 mmoles) of 1,3-dibromo-5,5-dimethylhydantoin are added to the mixture and stirred for 3 hours a. 20 ° C. Cool to 0 ° C, add an additional 1.0 g (3.5 mmoles) of dibromohydantoin and stir at 20 ° C for 2 hours. Pour the mixture into 400 g of ice, basify with NH40H (aqueous) concentrated at 0 ° C and the resulting solid is collected by filtration. The solid is washed with 300 ml of water, stirred in 200 ml of acetone and filtered to provide 19.79 g (85.6% yield) of the product, m.p. = 236 - 237 ° C, Mass spectrum: MH + = 584 (Cl). Elemental Analysis: Calculated - C, 45.11; H, 3.44; N, 7.17; found - C, 44.95; H, 3.57; N, 7.16 Stage C g (447 mmoles) of Fe filings, 10 g (90 mmoles) of CaCl 2 and a suspension of 20 g are combined (34.19 mmoles) of the product from step B in 700 ml of 90:10 EtOH / water at 50 ° C. The mixture is heated to reflux overnight, filtered through CeliteHR and the filter cake is washed 2 x 200 ml of hot EtOH. The filtrate and washings are combined and concentrated in vacuo to a residue. The residue is extracted with 600 ml of CH2C12 / washed with 300 ml of water and dried over MgSO4. Filter and concentrate in vacuo to a residue, and then chromatograph (silica gel, 30% EtOAc / CH2Cl2) to provide 11.4 g (60% yield) of the product, m.p. = 211 - 212 ° C, Mass spectrum: MH + = 554 (Cl). Elementary Analysis: Calculated - C, 47.55; H, 3.99; N, 7.56; found - C, 47.45; H, 4.31; N, 7.49.

Stage D: Slowly add (in portions) 20 g (35.9 mmoles) of the product from step C to a solution of 8 g (116 mmoles) of NaNO, in 120 ml of HCl (aqueous) concentrated at -10 ° C. The resulting mixture is stirred at 0 ° C for 2 hours, and then 150 ml (1.44 moles) of 50% H3P02 are slowly added (dropwise) at 0 ° C over a period of 1 hour. It is stirred at 0 ° C for 3 hours, and then poured into 600 g of ice and basified with concentrated NH.sub.3 H (aqueous). Extract with 2 X 300 mL of CH2C12, dry the extracts over MgSO4 and then filter and concentrate in vacuo to a residue. Chromatograph the residue (silica gel, 25% EtOAc / hexanes) to provide 13.67 g (70% yield) of the product, m.p. = 163 - 165 ° C, Mass spectrum: Y¡H + = 539 (Cl). Elementary Analysis: Calculated - C, 48.97; H, 4.05; N, 5.22; found - C, 48.86; H, 3.91; N, 5.18.

Stage E: Combine 6.8 g (12.59 mmol) of the product from step D and 100 ml of concentrated HCl (aqueous) and stir at 85 ° C overnight. The mixture is cooled, poured into 300 g of ice and basified with concentrated NH40H (aqueous). Extract with 2 x 300 ml of CH2C12 and then dry the extracts over MgSO4. Filter, concentrate in vacuo to a residue and then chromatograph (silica gel, 10% MeOH / EtOAc + 2% NH40H (aqueous)) to give 5.4 g (92% yield) of the title compound, m.p. = 172-174 ° C, Mass spectrum: MH "= 467 (BAR) Elemental Analysis: Calculated - C, 48.69; H, 3.65; N, 5.97; found - C, 48.83; H, 3.80; N, 5.97.

Preparatory Example 3 Combine 16.6 g (0.03 mole) of the product from preparative example 3, step D, with a 3: 1 solution of CH3CN and water (212.65 ml of CH3CN and 70.8 ml of water) and stir the resulting suspension overnight at room temperature. ambient. 32.833 g (0.153 mol) of NaI04 and then 0.31 g (2.30 mmol) of Ru02 are added and stirred at room temperature to provide 1.39 g (69% yield) of the product. (The addition of RuO is accompanied by an exothermic reaction and the temperature rises from 20 ° to 30 ° C). The mixture is stirred for 1.3 h (the temperature returns to 25 ° C after about 30 min), then it is filtered to remove the solids and the solids are washed with CH2C12. The filtrate is concentrated in vacuo to a residue and the residue is dissolved in CH2C12. Filter to remove insoluble solids and wash the solids .ccn CH2C12. The filtrate is washed with water, concentrated to a volume of about 200 ml and washed with bleach, and then with water. Extract with 6N HCl (aqueous). The aqueous extract is cooled to 0 ° C and 50% NaOH (aqueous) is added slowly to adjust the pH = 4 while maintaining the temperature < 30 ° C. It is extracted twice with CH 2 Cl 2, dried over MgSO 4 and concentrated in vacuo to a residue. The residue is suspended in 20 ml of EtOH and cooled to 0 ° C. The resulting solids are collected by filtration and the solids are dried in vacuo to provide 7.95 g of the product. 1 H NMR (CDC13, 200 MHz): 8.7 (s, 1H); 7.85 (m, 6H); 7.5 (d, 2H); 3.45 (m, 2H); 3.15 (m, 2H).

Preparatory Example 4 Stage A: g (38.5 mmol) of the 4- (8-chloro-3-bromo-5,6-dihydro-11H-benzo [5,6] cyclohepta [1,2-b] pyridine-ethyl ester are combined. ylidene) -1-piperidine-1-carboxylic acid and 150 ml of H, S04 concentrated at -5 ° C, then 3.89 g (38.5 mmoles) of KN03 were added and stirred for 4 hours. The mixture is poured into 3 1 of ice and basified with 50% NaOH (aqueous). Extract with CH2C12, dry over MgSO4 and then filter and concentrate in vacuo to a residue. The residue is recrystallized from acetone to provide 6.69 g of the product. X H NMR (CDC13, 200 MHz): 8.5 (s, 1 H), 7.75 (s, 1 H); 7.6 (s, 1H); 7.35 (s, 1H); 4.15 (c, 2H); 3.8 (m, 2H); 3.5-3.1 (m, 4H), - 3.0-2.8 (m, 2H); 2.6-2.2 (m, 4H); 1.25 (t, 3H).

Stage B: 6.69 g (13.1 mmol) of the product of step A and 100 ml of 85% EtOH / water are combined, and then 0.66 g (5.9 mmol) of CaCl2 and 6.56 g (117.9 mmol) of Fe are added and the mixture is heated reflux overnight. The hot reaction mixture is filtered through Celite ™ and the filter cake is rinsed with hot EtOH. The filtrate is concentrated in vacuo to provide 7.72 g of the product. Mass Spectrum MH + = 478.0.

Stage C: 7.70 g of the product from step B and 35 ml of HOAc are combined, then 45 ml of a Br2 solution in HOAc are added and the mixture is stirred at room temperature overnight. Add 300 ml of 1 N NaOH (aqueous), then 75 ml of 50% NaOH (aqueous) and extract with EtOAc. The extract is dried over MgSO4 and concentrated in vacuo to a residue. The residue is chromatographed (silica gel, 20% -30% EtOAc / hexane) to give 3.47 g of the product (together with another 1.28 g of partially purified product). Mass spectrum MH + = 555.9. X H NMR (CDC13, 300 MHz): 8.5 (s, 1H); 7.5 (s, 1H); 7.15 (s, 1H); 4.5 (s, 2H); 4.15 (m, 3H); 3.8 (broad s, 2H); 3.4-3.1 (m, 4H); 9-2.75 (m, 1H); 2.7-2.5 (m, 2H); 2.4-2.2 (m, 2H); 1.25 (m, 3H).

Stage D: Combine 0.557 g (5.4 mmol) of t-butylnitrite and 3 ml of DMF, and heat the mixture to 60 ° -70 ° C. A mixture of 2.00 g (3.6 mmol) of the product from step C and 4 ml of DMF is added slowly (dropwise), then the mixture is cooled to room temperature. Another 0.64 ml of t-butylnitrite is added at 40 ° C and the mixture is re-heated at 60 ° -70 ° C for 0.5 h. It is cooled to room temperature and the mixture is poured into 150 ml of water. Extract with CH2C12, dry the extract over MgSO4 and concentrate in vacuo to a residue.

The residue is chromatographed (silica gel, 10% -20% EtOAc / hexane) to give 0.74 g of the product. Mass spectrum MH "= 541.0 XH NMR (CDC13, 200 MHz): 8.52 (s, 1H), 7.5 (d, 2H), 7.2 (s, 1H), 4.15 (c, 2H), - 3.9-3.7 ( m, 2H), 3.5-3.1 (m, 4H), 3.0-2.5 (m, 2H), 2.4-2.2 (m, 2H), 2.1-1.9 (m, 2H), -1.26 (t, 3H).

Stage E: Combine 0.70 g (1.4 mmol) of the product of step D and 8 ml of concentrated HCl (aqueous) and heat the mixture to reflux overnight. 30 ml of 1 N NaOH (aqueous) is added, then 5 ml of 50% NaOH (aqueous) and extracted with CH2C12. Dry the extract over MgSO4 and concentrate in vacuo to provide 0.59 g of the title compound. Mass spectrum M + 468.7. p.f. = 123.9-124.2 ° C.

The title compound can be separated by the methodology of Preparative Example 3 to prepare the corresponding 11-ketone having the 3,10-dibromo-8-chloro substituents.

Preparatory Example 5 Stage A: 40.0 g (0.124 mol) of the initial ketone and 200 ml of H2SO4 are combined and cooled to 0 ° C. 13.78 g (0.136 mol) of K_T03 are slowly added over a period of 1.5 h, and then warmed to room temperature and stirred overnight. The reaction is treated using substantially the same procedure as described for preparative example 2, step A. It is chromatographed (silica gel, 20%, 30%, 40%, 50% EtOAc / hexane, and then 100% EtOAc ) to provide 28 g of the 9-nitro product together with a smaller amount of the 7-nitro product and 19 g of a mixture of the 7-nitro and 9-nitro compounds.

Stage B: 28 g (76.2 mmol) of the 9-nitro product from step A, 400 ml of 85% EtOH / water, 3.8 g, (34.3 mmol) of CaCl2 and 38.28 g (0.685 mol) of Fe are reacted using substantially the Same procedure as described for preparative example 2, step C, to provide 24 g of the product.

Stage C: 13 g (38.5 mmoies) of the product from step B, 140 ml of HOAc are combined and a solution of 2.95 ml (57.8 mmoles) of Br2 in 10 ml of HOAc is added slowly over a period of 20 min. The reaction mixture is stirred at room temperature and then concentrated in vacuo to a residue. CH2C12 and water are added, and then adjusted to pH = 8-9 with 50% NaOH (aqueous). The organic phase is washed with water, then with brine and dried over Na 2 SO 4. Concentrate in vacuo to provide 11.3 g of the product.

Stage D: 100 ml of concentrated HCl (aqueous) is cooled to 0 ° C, then 5.61 g (81.4 mmoles) of NaNO are added, and the mixture is stirred for 10 minutes. Slowly add (in portions) 11.3 g (27.1 mmoles) of the product from step C and stirred into the mixture at 0 ° -3 ° C for 2.25 h. 180 ml of H3P02 (aqueous) 50% are added slowly (dropwise) and the mixture is allowed to stand at 0 ° C overnight. Slowly add (dropwise) 150 ml of 50% NaOH for 30 min, to adjust to pH = 9, and then extract with CH2C12. The extract is washed with water, then with brine and dried over Na 2 SO 4. Concentrate in vacuo to a residue and chromatograph (silica gel, 2% EtOAc / CH, Cl2) to provide 8.6 g of the product.

Example 1 Stage A The compound of Preparative Example 1 (0.95 g, 1.9 mmol) is dissolved in 34 ml of toluene. Triethylamine (1 ml) and ethyl chloroformate (1.82 ml, 10 eguivalents) are added and the reaction mixture is refluxed for 2 hours. The reaction mixture is cooled to room temperature and evaporated to an oil. The oil is chromatographed on silica gel using 15 to 20% ethyl acetate / hexanes to obtain 0.77 g of 4- (3,10-dibromo-8-chloro-6,11-dihydro-11-hydroxy-5H-). ethyl benzo [5,6] cyclohepta [1, 2-b] pyridin-11-yl) -1-piperidinecarboxylate. EMBAR (M + H) 559. * Stage B: The compound of Preparative Example 2 (0.37 g) is dissolved in 5 ml of concentrated hydrochloric acid and refluxed for 18 hours. The mixture is evaporated to a brown solid of compound 4- (3,10-dibromo-8-chloro-6,11-dihydro-l-hydroxy-5H-benzo [5,6] cyclohepta [1,2-b] - pyridin-11-yl) -1-piperidine and used without chromatography.

Stage C: The compound of Preparative Example 3 (100 mg, 0. 206 mmoles) is dissolved in 2 ml of N, N-dimethylformamide. 4-Pyridylacetic acid-N-oxide (126 mg, 0.82 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (DEC) (0.079 mg, 0.5 mmol)), 1-hydroxybenzotriazole (HOBt) ( 0.056 g, 0.5 mmol) and N-methylmorpholine (0.23 ml, 2.0 mmol) and the reaction mixture is stirred at room temperature. After 24 hours, the reaction mixture is added to brine and extracted with 3 X 15 ml of ethyl acetate. The combined ethyl acetate washes are combined and the solvent is evaporated under vacuum to provide a gum. The gum is subjected to flash chromatography on silica gel using 10% methanol / methylene chloride as the eluent to obtain 0.076 g of 4- (3,10-dibromo-8-chloro-6,1-dihydro-11-hydroxy). 5H-benzo [5, 6] cyclo epta [1, 2-b] -pyridin-11-yl) -1- (4-piperidinylacetyl) piperidine NI-oxide. EMBAR (M + H) = 622.

Example 2 Stage A: The procedure of example 1 above is followed by substituting 4-pyridylacetic acid-N-oxide with N-Boc-4-piperidineacetic acid to obtain 4- (3, 10-dibromo-8-chloro-6,11-dihydro-11-) hydroxy-5H-benzo [5,6] cyclohepta [1,2-b] -pyridin-11-yl) - (N-BOC-4-piperidinacetyl) iperidine in 85% yield. High resolution MS: observed = 712.0975.

Stage B: The compound from step A above (0.21 g) is dissolved in 50% trifluoroacetic acid / methylene chloride and stirred for 1 hour. The reaction mixture is evaporated to obtain an oil which is dissolved in 2 ml of methylene chloride to give 4- [2- [4- (3,10-dibromo-8-chloro-6,11-dihydro-11- hydroxy-5H-benzo [5,6] cyclohepta [1, 2-b] -pyridin-11-yl) -1-piperidinyl] -2-oxoethyl] -1-piperidine.

Stage C: Trimethylsilylisocyanate (234 μL, 1.47 mmol) is added and the reaction mixture is stirred at room temperature for 15 hours. The solvent is evaporated and the crude product is chromatographed on silica gel using 7.5% methanol / methylene chloride to obtain 100 mg, 62% of 4- [2- [4- (3,10-dibromo-8-chloro-6 , 11-dihydro-11-hydroxy-5H-benzo [5,63 cyclohepta [1,2- b] -pyridin-11-yl) -1-piperidinyl] -2-oxoethyl] -i-piperidinecarboxamide (pipepdine). ? M high resolution: observed = 655.0509.

Example 3 The compound of Example 2, step A above (0.069 g, 0.113 mmole) is dissolved in 2 ml of N, N-dimethylformamide. Sodium carbonate (0.036 g, 0. 34 mmole) and bromoacetamide (0.023 g, 0.17 mmole) and the reaction mixture is stirred at room temperature for 24 hours. The mixture is added to brine and extracted with ethyl acetate. The ethyl acetate layer is dried over magnesium sulfate, filtered and evaporated to obtain a crude solid. The crude solid is chromatographed on silica gel using 5% methanol / methylene chloride to obtain 4- [2- [4- (3, 10-dibromo-8-chloro-6,1, -dihydro-11-hydroxy). 5H-benzo [5,6] cyclohepta [1,2-b] -pyridin-11-yl) -1-piperidinyl] -2-oxoethyl] -1-piperidineacetamide. High resolution MS: observed = 669.0666. The following compounds can be prepared using the procedure of Example 1 above by substituting the following carboxylic acids for 4-pyridylacetic acid-N-oxide.

Carboxylic acid T = in Formula 1. 0 H00CCH2C0NH2 -0CCH2C0NH2 H00CCH, C0.H -0CCH, C02H x 0, SO ,, S or SMe X O, SO ,, S or SMe R = CONH2, COCH3 R = CONH2, C0CH3 R1 = CONH2 / COCH3 R1 = CONH2, COCH3 HOOCCH2NH2 -OCCH2NH2 HOOCCH2-COOH -OCCH.-COOH ESSAYS FPT IC50 (inhibition of farnesii protein transferase, enzyme assay in vitro) and COS Cell CIS0 (cell-based assay) were determined following the assay procedures described in WO 95/10516, published on April 20, 1995. GGPT CIS0 (inhibition of geranylgeranyl protein transferase enzyme assay in vitro), Mat cell assay and antitumor activity (anti-tumor studies in vivo) can be determined by the assay met described in WO 95/10516. The description of WO 95/10516 is incorporated herein by reference thereto. Additional assays can be carried out by following essentially the same procedure as described above, but with substitution of an alternative indicator of the tumor cell lines instead of the T24-BAG cells. Assays can be carried out using either DLD-1-BAG human colon carcinoma cells expressing an activated K-ras gene or human colon carcinoma cells SW620-BAG expressing an activated K-ras gene. Using other tumor cell lines known in the art, the activity of the compounds of this invention against other types of cancer cells can be demonstrated.

Ensavo in Aerar Suave: The independent growth of the anchor is characteristic of tumorigenic cell lines. Human tumor cells are suspended in growth medium containing 0.3% agarose and an indicated concentration of farnesyl transferase inhibitor. The solution is superimposed on solidified growth medium with 0.6% agarose containing the same concentration of farnesyl transferase inhibitor as the upper layer. After the top layer solidifies, the plates are incubated for 10-16 days at 37 ° C under C02 5% to allow the growth of the colonies. After incubation, the colonies are stained by overlaying agar with a solution of MTT, (3- [4,5-dimethyl-thiazol-2-yl] -2,5-diphenyltetrazolium bromide, thiazolyl blue) (1) mg / ml in PBS). The colonies are counted and the IC50 can be determined. The compounds of examples 1 stage C, 2 stage A, 2 stage B, 2 stage C and 3 have a FPT IC50 within the range of < 0.002 μM and 0.042 μM. To prepare pharmaceutical compositions from the compounds described by this invention, the inert and pharmaceutically acceptable carriers can be solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, sacks and suppositories. The powders and tablets may be comprised from about 5 to about 70 percent active ingredient. Suitable solid carriers are known in the art, for example magnesium carbonate, magnesium stearate, talc, sugar, lactose. Tablets, powders, sacks and capsules can be used as solid dosage forms suitable for oral administration. To prepare suppositories, a wax with a low melting point such as a mixture of fatty acid giicerides or cocoa butter first melts, and the active ingredient is dispersed homogeneously therein by agitation. The molten homogeneous mixture is then poured into molds of suitable dimension, allowed to cool and subsequently solidify. Liquid form preparations include solutions, suspensions and emulsions. As an example, water or water-propylene glycol solutions for parenteral injection may be mentioned. Liquid form preparations may also include solutions for intranasal administration. Aerosol preparations suitable for inhalation may include solutions and solids in powdered 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 designed to be converted, soon after use, to liquid form preparations for oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions. The compounds of the invention can also be transdermally derivable. The transdermal compositions can take the form of creams, lotions, aerosols and / or emulsions and can be included in a transdermal patch of the reservoir type matrix as is conventional in the art for this purpose. Preferably, the compound is administered orally. Preferably, the pharmaceutical preparation is in a unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component, for example, an effective amount to obtain the desired purpose. The amount of active compound in a unit dose of preparation can vary or be adjusted from about 0.1 mg to 1000 mg, more preferably from about 1 mg to 300 mg, according to the particular application. The current dosage used may vary depending on the requirements of the patient and the severity of the condition to be treated. The determination of the appropriate dosage for a particular situation is within the skills of the technique. Usually, the treatment starts with smaller dosages which are less than the optimum dose of the compound. Subsequently, the dosage is increased by small increments until the optimum effect is reached under the circumstances. For convenience, the total daily dosage can 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 physician considering factors such as the age, condition and size of the patient as well as the severity of the symptoms that are going to be treated. A typical recommended dosage regimen is oral administration of 10 mg to 2000 mg / day, preferably 10 to 1,000 mg / day, in 2 to 4 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 limited by the examples provided.

Examples of pharmaceutical dosage form EXAMPLE A Tablets Method of Preparation Items 1 and 2 are mixed in a suitable mixer for 10-15 minutes. The mixture is granulated with item number 3. The moistened granules are milled through a coarse mesh (for example 0.63 cm, 1/4") if necessary.The moistened granules are dried.The dry granules are sieved if It is mixed and mixed with item number 4 and mixed for 10-15 minutes Add item number 5 and mix for 1-3 minutes Compress the mixture to the appropriate size and weight in a suitable tabletting machine.

EXAMPLE B Capsules Manufacturing Method Articles 1, 2 and 3 are mixed in a suitable mixer for 10-15 minutes. Item number 4 is added and mixed for 1-3 minutes. The mixture is filled into suitable two-piece hard gelatin capsules on a suitable encapsulating machine. Although the present invention has been described in conjunction with the specific embodiments set forth in the foregoing, many alternatives, modifications and variations thereto will be apparent to those ordinarily skilled in the art. All of such alternatives, modifications and variations are considered to be within the spirit and scope of the present invention.

Claims (12)

REI INDICATION?

1. A compound of the formula: or a pharmaceutically acceptable salt or solvate thereof, wherein: a represents N or NO ", - R1 and R3 are the same or different and each represents halo, - R2 and R4 are the same or different and each is selected from H and halo, with the proviso that at least one of R2 and R4 is H; T is a substituent selected from S02R or Z is O or S; n is zero or an integer from 1 to 6; R is alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl or N (R5) 2; Rs is H, alkyl, aryl, heteroaryl or cycloalkyl.

2. . The compound of claim 1, which is selected from: (1) wherein a, T, R1 and R3 are as defined in claim 1; (2) wherein a, T, R1, R3 and R4 are as defined in claim 1, - (3) wherein a, T, R1, R2 and R4 are as defined in claim 1; (4) wherein a, T, R1, R2, R3 and R4 are as defined in claim 1; or (5) wherein a, T, R1, R2, R3 and R4 are as defined in claim i.

3. The compound of claim 2, wherein: (1) for the compound of formula 1.0a, R1 is bromine and R3 is chlorine, - (2) for the compound of formula 1.1, R1 is bromine R3 is chlorine and R4 is bromine, - and (3) for the compound of formula 1.2, R1 is bromine, R2 is bromine and R3 is chlorine.

4. The compound of claim 3, wherein, for the compounds of formulas 1.0a, 1.1 and 1.2, T is -S02 methyl or a group wherein R is a 3-pyridinyl N-oxide, 4-pyridinyl N-oxide, 4-piperidinyl, 3-piperidinyl or 3-pyrrolidinyl group, wherein the 4-piperidinyl, 3-piperidinyl or 3-pyrrolidinyl groups may be substituted in the piperidinyl or pyrrolidinyl nitrogen with a R9 group; R9 is selected from -C (O) N (R10) 2, -CH2C (O) N (R10) 2, -S02R10, -SO2N (R10) 2, -CiO)! 11, -CioJOR11, alkyl, aryl, aralkyl, cycloalkyl, heterocycloalkyl or heteroaryl; each R10 independently represents H, alkyl, aryl or aralkyl; and R11 is alkyl, aryl, aralkyl, heteroaryl or heterocycloalkyl.

5. The compound of claim 4, wherein, for the compounds of formulas 1.0a, 1.1 and 1.2, the carbon at the C-ll position is in the R configuration.

6. The compound of claim 4, wherein, for the compounds of formulas 1.0a, 1.1 and 1.2, the carbon at the C-ll position is in the S configuration.

A compound of claim 1, which has the formula: ; 10

8. The use of a compound of claim 1 for the manufacture of a medicament for treating tumor cells, which express an activated ras oncogene.

9. The use of claim 8, wherein the treated tumor cells are pancreatic tumor cells, lung cancer cells, myeloid leukemia tumor cells, thyroid follicular tumor cells, myelodysplastic tumor cells, epidermal carcinoma tumor cells, carcinoma tumor cells, bladder, colon tumor cells, breast tumor cells and prostate tumor cells.

10. The use of a compound of claim 1, for the manufacture of a medicament for treating tumor cells, wherein the Ras protein is activated as a result of the oncogenic mutation in genes other than the Ras gene.

11. The use of a compound of claim 1 for the manufacture of a medicament for inhibiting farnesyl protein transferase.

12. A pharmaceutical composition for inhibiting farnesyl protein transferase, comprising an effective amount of the compound of claim 1 in combination with a pharmaceutically acceptable carrier. SUMMARY OF THE INVENTION Novel compounds of formula (1.0) or a pharmaceutically acceptable salt or solvate thereof are described in which: a represents N, NO ", R1 and R3 are the same or different and each represents halo, R2 and R4 are the same or different and each one is selected from H and halo, with the proviso that at least one of R2 and R4 is H; T is a substituent selected from SO2 R or (A); Z is O or S; n is zero or an integer from 1 to 6; R is alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, or N (R5) 2; R5 is H, alkyl, aryl, heteroaryl or cycloalkyl; Methods for inhibiting famesyl protein transferase and methods for treating tumor cells are also described; novel compounds of formula (1.0) or a pharmaceutically acceptable salt or solvate thereof are described in which: a represents N, NO "; R1 and R3 are the same or different and each represents halo; R2 and R4 are equal N. or different and each is selected from H and halo, with the proviso that at least one of a substituent selected from SO2 R or (A); Z is O o; p. is zero or an integer from 1 to 6; R is alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkio, heterocycloalkyl or N (R5) 2; R5 is H, alkyl, aryl, heteroaryl or cycloalkyl; methods for inhibiting famesii protein transferase and methods for treating tumor cells are also described. DR Iss P99 / 1698F