MXPA99012062A - Benzo(5,6)cycloheptapyridine cyclic ureas and lactams useful as farnesyl protein transferase inhibitors - Google Patents

Abstract

Compounds of formula (I) useful for inhibiting Ras function and therefore inhibiting or treating the abnormal growth of cells are disclosed, or a pharmaceutically acceptable salt or solvate thereof, wherein:R and R2 are halo;R1 and R3 are H and halo, provided that at least one is H;W is N, CH, or C when the double bond is present at the C-11 position;R4 is -(CH2)n-R5 or (II);R5 is (III);R6 is R5 or (IV);Z1 and Z2 are indepen dently selected from the group consisting of=O and=S;n is 1-6;and n1 is 0 or 1.

Description

UREAS AND LACTA AS CYCLIC OF BENZO (5,6) CICLOHEPTAPlRIDINA. USEFUL AS FARNESIL-PROTEIN TRANSFERASE INHIBITORS BACKGROUND OF THE INVENTION The international publication number WO95 / 10516, published on April 0, 1995, describes compounds of the formula: wherein R can be heterocycloalkyl attached to the carbon of the group -C (= Z) -by a heterogeneous atom, substituted piperidinyl or substituted piperidinylmethyl. It is mentioned that the compounds are useful for inhibiting farnesyl protein transferase.

BRIEF DESCRIPTION OF THE INVENTION The compounds of the present invention are represented by the formula I: or an N-oxide thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: R1 and R2 are independently selected from halogen; R1 and R3 are independently selected from the group consisting of H and halogen, as long as at least R1 and R3 are H; W is N, CH or C when the double bond is present at the C11 position; Z1 and Z2 are independently selected from the group consisting of = O and = S; n is 1-6; and n-i is 0 or 1. In the com ponents of the invention, preferably R is Br, R 2 is halogen and R 1 is halogen; or R is Br; R2 is halogen and R3 is halogen; or R is Br, R 2 is halogen and R 1 and R 3 are each H. R 2 is preferably Br or Cl. When R 1 or R 3 is halogen, it is preferably Br or Cl. Z 1 is preferably = O. Z2 is preferably = O. W is preferably CH. The values that are preferred for n are n 1-3. R5 and R6 are preferably When R4 is ni is preferably 1 and the resulting piperidinyl group is preferably linked to the methylene at the carbon ring member at position 4. The compounds of this invention: (i) potently inhibit farnesyl- transferase protein, but not geranylgeranyl transferase I, in vitro; (ii) block the phenotypic change induced by a form of transforming Ras which is a farnesyl receptor but not by a form of transforming Ras designed to be a geranylgeranyl receptor; (iii) block the intracellular processing of Ras which is a farnesyl receptor but not Ras designed to be a geranylgeranyl receptor; and (iv) they block the abnormal growth of cells in induced culture by transforming Ras. The compounds of this invention inhibit farnesyl protein transferase and farnesylation of the Ras oncogenic protein. This invention further provides a method for inhibiting ras famesyl-protein transferase in mammals, especially humans, by administering 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 below. The compounds of this invention inhibit famesyl transferase and farnesylation of the Ras oncogenic protein. This invention further provides a method for inhibiting Ras famesyl transferase in mammals, especially humans, by administering an effective amount of the tricyclic compounds described above. The administration of the compounds of this invention to patients to inhibit famesyl transferase is useful in the treatment of the 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 cell growth independent of normal regulatory mechanisms (eg, 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 the aberrant activation of Ras 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) that requires such treatment. In particular, this invention provides a method for inhibiting or treating the growth of tumors expressing an activated Ras oncogene by administering an effective amount of the compounds described above. Examples of tumors that can be inhibited or treated include, but are not limited to, breast cancer, prostate cancer, 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 (e.g., acute myelogenous leukemia (AML)), follicular thyroid cancer, myelodysplastic (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 - that is, the Ras gene itself is not activated by the mutation to an oncogenic form - said inhibition or treatment being achieved by administering an effective amount of the tricyclic compounds described herein, to a mammal (e.g., a human) that requires 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 (eg, neu, src, abl, Ick and fyn), can be inhibited by tricyclic compounds described in the present. The tricyclic compounds useful in the methods of this invention inhibit or treat abnormal cell growth. Without wishing to be bound by theory, it is believed that these compounds could function by inhibiting the function of the G protein, such as ras p21, by blocking the isoprenylation of the G protein, 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 exhibit 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; Bu represents butyl; Et represents ethyl; It represents me methyl; Ph represents phenyl; and halogen represents fluorine, chlorine, bromine and iodine. The following solvents and reagents may be mentioned herein by means of the indicated abbreviations: tetrahydrofuran (THF); Ethanol (EtOH); methanol (MeOH); acetic acid (HOAc or AcOH); ethyl acetate (EtOAc); N, N-dimethylformamide (DMF), trifluoroacetic acid (TFA); trifluoroacetic anhydride (TFAA); 1-hydroxybenzotriazole (HOBT); m-chloroperbenzoic acid (MCPBA); triethylamine (Et3N); diethyl ether (Et20); ethyl chloroformate (CICO2Et) and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (DEC).

The representative structures of formula I with respect to W and the optional double bond are as follows: The lines drawn on the ring systems indicate that the indicated bond can be attached to any of the substitutable ring carbon atoms. Certain compounds of the invention can exist in different isomeric forms (e.g., enantiomers and diastereomers). The invention contemplates all such isomers in both pure and mixed forms, including racemic mixtures. Enol forms are also included. Certain tricyclic compounds 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, gold and silver salts. Also contemplated are salts formed with pharmaceutically acceptable amines, such as ammonia, alkylamines, hydroxyalkylamines, N-methylglucamine and the like. Certain basic tricyclic compounds also form pharmaceutically acceptable salts, for example, acid addition salts. For example, pyrido-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 technique. 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 suitably diluted aqueous base solution, such as dilute aqueous NaOH, potassium carbonate, ammonia and sodium bicarbonate. The free base forms are different from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but the acidic and basic salts are otherwise equivalent to their respective free base forms for the purposes of invention. All such acidic and basic salts are designed to be pharmaceutically acceptable salts within the scope of the invention, and all acidic and basic salts are considered equivalent to the free forms of the corresponding compounds for the purposes of the invention. The compounds of the invention can be made by methods described in the examples below, and using the methods described in WO 95/10516 - see, for example, the methods for preparing compounds of the formula 400.00. The compounds of the invention in which Z1 and Z2 are = O can be prepared by reacting a compound of the formula II or III wherein all other substituents are as defined for formula I, with an acid of the formula HOOC- (CH2) n-NHR7 or HOOC- (CH2) n3-NHR7, respectively, wherein n is as defined above and R7 is an amino protecting group such as t-butoxycarbonyl (BOC). The reaction is carried out using normal amide coupling conditions, for example, the reaction can be carried out at room temperature, in an inert solvent such as DMF, in the presence of a condensing agent such as hydrochloride of 1- ( 3-dimethylaminopropyl) -3-ethylcarbodiimide, a base such as N-methylmorpholine and an activating agent such as 1-hydroxybenxotriazole. The protecting group of R7 is then removed, for example, by treatment with trifluoroacetic acid, to obtain the corresponding amine of the formula HA or I HA: To prepare the compounds of the formula I wherein R5 or R6 comprises a cyclic lactam, a compound of the formula NA or IIIA is reacted with 4-bromobutyryl chloride or 4-bromovaleryl chloride, followed by cyclization with a reagent such as NaH. The compounds of the formula I wherein R 5 or R 6 comprise a cyclic urea are prepared in a similar manner by reacting an amine of the formula NA or I HA with 2-bromoethyl isocyanate or 3-chloropropyl isocyanate, followed as above by cyclization with a reagent such as NaH.

Alternatively, an amine of the formula HA or I HA can be reacted with lactam substituted acetate under normal amide coupling conditions such as those described above. When Z1, or Z1 and Z2 represent sulfur, a compound of formula I wherein Z1, or Z1 and Z2 is oxygen, is reacted with P2Ss, Lawesson's reagent or another reagent capable of introducing sulfur instead of oxygen. The reaction can take place at elevated temperature in pyridine, toluene or other suitable solvents. For the compounds wherein Z1 and Z2 are different, the conversion of oxygen to sulfur can be carried out before the starting materials (ie, the compounds of the formula I HA and the alkanoyl chloride or isocyanate) are reacted . Compounds of formula I comprising a pyridyl N-oxide in ring I of the tricyclic portion can be prepared by methods well known in the art. For example, the compound of formula II wherein W is C or CH can be reacted with MCPBA in a suitable organic solvent, for example, CH2Cl2 (normally anhydrous) at a suitable temperature, to obtain an N-oxide of the formula Ha Generally, the organic solvent solution of formula II is cooled to about 0 ° C before adding MCPBA. The reaction is then allowed to warm to room temperature during the reaction period. The desired product can be recovered by normal separation means, for example, the reaction mixture can be washed with an aqueous solution of a suitable base, for example, NaHCOs or saturated NaOH (for example, 1 N NaOH), and then dried over MgSO anhydrous. The solution containing the product can be concentrated in vacuo, and the product can be purified by normal means, for example, by chromatography using silica gel (e.g., column chromatography by vaporization). The compounds of the formula II are prepared by methods known in the art, for example by methods described in WO 95/10516 in E.U.A. 5,151, 423 and those described below. The compounds of the formula II wherein the C-3 position of the pyridine ring in the tricyclic structure is replaced by bromine can also be prepared by a process comprising the following steps: (a) reacting an amide of the formula wherein R 11a is Br, R 5a is hydrogen and R 6a is C 1 -C 6 alkyl, aryl or heteroaryl; R5a is C6-C6 alkyl, aryl or heteroaryl and R6a is hydrogen; R5a and R6a are independently selected from the group consisting of C6-C6 alkyl and aryl; or R5a 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 heterogeneous portion selected from the group consisting of -O- and -NR9a- , wherein R9a is H, C -? - C6 alkyl or phenyl; with a compound of the formula wherein R1a, R2a, R3a and R4a are independently selected from the group consisting of hydrogen and halogen, 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 (i) POCI3 to obtain a cyano compound of the formula (ii) or DIBALH to obtain an aldehyde of the formula (c) reacting the cyano compound or the aldehyde with a piperidine derivative of the formula wherein L is a leaving group selected from the group consisting of Cl and Br, to obtain an aldehyde or an alcohol of the following formula, respectively: (d) (i) cyclizing the aldehyde with CF3SO3H to obtain a compound of formula II wherein the dotted line represents a double bond; or (d) (ii) cyclizing the alcohol with polyphosphoric acid to obtain a compound of formula II wherein the dotted line represents an individual bond. The methods for preparing the compounds of the formula II described in WO 95/10516, E.U.A. 5,151, 423 and described below employ a cyclic ketone intermediate. These intermediaries of the formula R 1 wherein R 11, R 1a, R 2a, R 3a and R 4a are independently selected from the group consisting of hydrogen and halogen, can be prepared by the following process comprising: (a) reacting a compound of the formula (i) with an amine of the formula NHR5aR6a, wherein R5a and R6a 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-i-Cß or cycloalkyl of OrCß, in the presence of a palladium and carbon monoxide catalyst to obtain the ester of the formula R followed by reaction of the ester with an amine of the formula NHR5aR6a to obtain the amide; (b) reacting the amide with an iodo-substituted benzyl compound of the formula wherein R1a, R2a, R3a, R4a 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 alkyl, aryl or heteroaryl of C? -C8, and L is Br or Cl, provided that prior to cyclization, the compounds in which R5a or R6a is hydrogen are reacted with an appropriate N-protecting group. The (+) isomers of the compounds of the formula II wherein X is CH can be prepared with high enantioselectivity using a process comprising transesterification catalyzed by enzyme. Preferably, a racemic compound of formula II, wherein X is C, the double bond is present and R3 is not H, is reacted with an enzyme such as Toyobo LIP-300 and an acylating agent such as sodium isobutyrate. trifluoroethyl; the resulting amide (+) is then hydrolyzed, for example by refluxing with an acid such as H2SO4, to obtain the corresponding optically enriched (+) isomer, wherein X is CH and R3 is not H. Alternatively, a racemic compound of formula II, wherein X is C, the double bond is present and R3 is not H, it is first reduced to the corresponding racemic compound of formula II where X is CH and then treated with the enzyme (Toyobo LIP-300 ) and acylating agent as described above to obtain the amide (+), which is hydrolysed to obtain the optically enriched (+) isomer. The compounds of the formula III can be prepared from the compounds of the formula II by procedures well known in the art, for example by reacting 1-Nt-butoxycarbonylpiperidinyl-4-acetic acid with the compound of the formula II under the normal amide coupling conditions described above. The compounds useful in this invention are exemplified by the following preparation examples, which should not be considered as limiting the scope of the description. Alternative mechanical routes and analogous structures within the scope of the invention may be apparent to those skilled in the art.

EXAMPLE OF PREPARATION 1 Step A .86 g (55. 9 mmol) of 4- (8-chloro-3-bromo-5,6-dihydro-11 H-benzo [5,6] cyclohepta [1,2-b] pyridine ethyl ester are combined. -11-ylidene) -1-piperidino-1-carboxylic acid and 250 mL of concentrated H2SO at -5 ° C, then 4.8 g (56.4 mmoles) of NaN 3 is added and stirred for 2 hours. The mixture is poured into 600 g of ice and made basic with concentrated NH OH (aqueous). The mixture is filtered, washed with 300 mL of water and then extracted with 500 mL of CH2CI2. The extract is washed with 200 mL of water, dried over MgSO4, then filtered and concentrated in vacuo to a residue. The residue is chromatographed (silica gel, 10% EtOAc / CH 2 Cl 2) to give 24.4 g (86% yield) of the product. P.f. = 165-167 ° C, Mass spectrum: MH + = 506, 508 (Cl). Elemental Analysis: Calculated C, 52.13; H, 4.17; N, 8.29 Found C, 52.18; H, 4.51; N, 8.16 Step B g (40.5 mmol) of the product from step A and 200 mL of concentrated H2SO are combined at 20 ° C, then the mixture is cooled to 0 ° C. 7.12 g (24.89 mmol) of 1,3-dibromo-5,5-dimethylhydantoin are added to the mixture and stirred for 3 hours at 20 ° C. Cool to 0 ° C, add 1.0 g (3.5 mmol) of the dibromohydantoin and stir at 20 ° C for 2 hours. The mixture is poured into 400 g of ice, made basic with concentrated NH 4 OH (aqueous) at 0 ° C, and the resulting solid is collected by filtration. The solid is washed with 300 mL of water, suspended 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 + = 586 (Cl). Elemental Analysis: Calculated- C, 45.11; H, 3.44; N, 7.17 Found-C, 44.95; H, 3.57; N, 7.16 Step C g (447 mmoles) of Fe fillers, 10 g (90 mmoles) of CaCl 2 and a 20 g suspension (34.19 mmoles) of the product of step B in 700 mL of 90:10 EtOH / water at 50 ° are combined. C. The mixture is heated to reflux overnight, filtered through Celite® and the filter cake is washed with 2 x 200 mL of hot EtOH. The filtrate is combined and washed, and concentrated in vacuo to a residue. The residue is extracted with 600 mL of CH2CI2, washed with 300 mL of water and dried over MgSO4-. It is filtered and concentrated in vacuo to a residue, then subjected to chromatography (silica gel, 30% EtOAc / CH2Cl2) to give 11.4 g (60% yield) of the product. P.f. = 211-212 ° C, Mass spectrum: MH + = 556 (Cl).

Elemental Analysis: Calculated-C, 47.55; H, 3.99; N, 7.56 Found-C, 47.45; H, 4.31; N, 7.49 Step D Slowly add (in portions) 20 g (35.9 mmol) of the product from step C to a solution of 8 g (116 mmol) of NaNO2 in 120 mL of concentrated HCl (aqueous) at -10 ° C. The resulting mixture is stirred at 0 ° C for 2 hours, then 150 mL (1.44 moles) of 50% H 3 PO 2 are added slowly (dropwise) at 0 ° C over a period of 1 hour. It is stirred at 0 ° C for 3 hours, then it is poured into 600 g of ice and made basic with concentrated NH 4 OH (aqueous). It is extracted with 2 x 300 mL of CH2Cl2, and the extracts are dried over MgSO4, then filtered and concentrated in vacuo to a residue. The residue is subjected to chromatography (silica gel, 25% EtOAc / hexanes) to give 13.67 g (70% yield) of the product. P.f. = 163-165 ° C, Mass spectrum: MH + = 541 (Cl).

Elemental Analysis: Calculated - C, 48.97; H, 4.05; N, 5.22 Found - C, 48.86; H, 3.91; N, 5.18 Step 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 made basic with concentrated NH 4 OH (aqueous). Extract with 2 x 300 mL of CH2Cl2, and then dry the extracts over MgSO4. Filter, concentrate in vacuo to a residue and then chromatograph (silica gel, 10% MeOH / EtOAc + 2% NH 4 OH (aq.)) To give 5.4 g (92% yield) of the compound of the title. P.f. = 172-174 ° C, Mass spectrum: MH + = 469 (FAB). Elemental Analysis: Calculated - C, 48.69; H, 3.65; N, 5.97 Found - C, 48.83; H, 3.80; N, 5.97 EXAMPLE OF PREPARATION 2 Step A 2.42 g of 4- (8-chloro-3-bromo-5,6-dihydro-11 H -benzo [5,6] cyclohepta [1,2-b] pyridin-11-ethylide is hydrolyzed. ) -1-piperidino-1-carboxylic acid by dissolving in concentrated HCl and heating at about 100 ° C for @ 16 hours. The mixture is cooled and then neutralized with 1M NaOH (aqueous). Extract with CH2CI2, dry the extracts over MgSO4, filter and concentrate in vacuo to give 1.39 g (69% yield) of the product.

Step B Combine 1 g (2.48 mmol) of the product of step A and 25 mL of dry toluene, add 2.5 mL of DIBAL to 1 M in toluene and heat the mixture to reflux. After 0.5 hours, add another 2.5 mL of 1 M DIBAL in toluene and heat at reflux for 1 hour. (The reaction is monitored by CCD using 50% MeOH / CH2Cl2 + NH4OH (aqueous)). The mixture is cooled to room temperature, 50 mL of 1 N HCl (aqueous) is added and stirred for 5 minutes. 100 mL of 1 N NaOH (aqueous) is added, then extracted with EtOAc (3 x 150 mL). The extracts are dried over MgSO 4, filtered and concentrated in vacuo to give 1.1 g of the title compound.

EXAMPLE OF PREPARATION 3 [racemic as well as isomers (+) and (-)] Step A Combine 16.6 g (0.03 mole) of the product of Preparation Example 1, 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. room temperature. 32.833 g (0.153 mol) of NalO4 and then 0.31 g (2.30 mmol) of Ru? 2 are added and the mixture is stirred at room temperature to give 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 hours (the temperature returns to 25 ° C after approximately 30 minutes), then it is filtered to remove the solids and the solids are washed with CH2Cl2. The filtrate is concentrated in vacuo to a residue and the residue dissolved in CH2Cl2. It is filtered to remove the insoluble solids and the solids are washed with CH2Cl2. The filtrate is washed with water, concentrated to a volume of about 200 mL and washed with bleach, then with water. Extract with HCl to 6N (aqueous). The aqueous extract is cooled to 0 ° C and 50% NaOH (aqueous) is added slowly to adjust to pH = 4, maintaining the temperature < 30 ° C. Extract twice with CH2CI2, dry over MgSO4 and concentrate 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 under vacuum to give 7.95 g of the product. 1 H NMR (CDCl 3> 200 MHz): 8.7 (s, 1 H); 7.85 (m, 6H); 7.5 (d, 2H); 3.45 (m, 2H); 3.15 (m, 2H).

Step B 21.58 g (53.75 mmol) of the product from step A and 500 mL of an anhydrous 1: 1 mixture of EtOH and toluene are combined, 1.43 g (37.8 mmol) of NaBH 4 is added and the mixture is heated at reflux for 10 minutes. The mixture is cooled to 0 ° C, 100 mL of water are added, and then it is adjusted to pH = 4.5 with 1 M HCl (aqueous) maintaining the temperature < 10 ° C. 250 mL of EtOAc are added and the layers are separated. The organic layer is washed with brine (3 x 50 mL) and then dried over Na2SO. Concentrate in vacuo to give a residue (24.01 g) and the residue is subjected to chromatography (silica gel, 30% hexane / CH2Cl2) to give the product. The impure fractions are purified by rechromatography. A total of 18.57 g of the product is obtained. 1 H NMR (DMSO-d 6) 400 MHz): 8.5 (s, 1 H); 7.9 (s, 1 H); 7.5 (d of d, 2H); 6.2 (s, 1 H); 6.1 (s, 1 H); 3.5 (m, 1 H); 3.4 (m, 1 H); 3.2 (m, 2H).

Step C 18.57 g (46.02 mmol) of the product from step B and 500 mL of CHCI3 are combined, then 6.70 mL (91.2 mmol) of SOCI2 are added, and the mixture is stirred at room temperature for 4 hours. A solution of 35.6 g (0.413 mol) of piperazine in 800 mL of THF is added over a period of 5 minutes and the mixture is stirred for 1 hour at room temperature. The mixture is heated to reflux overnight, then cooled to room temperature and the mixture is diluted with 1 L of CH 2 Cl. Wash with water (5 x 200 mL) and extract the aqueous wash with CHCl 3 (3 x 100 mL).

All organic solutions are combined, washed with brine (3 x 200 mL) and dried over MgSO. Concentrate in vacuo to a residue and then chromatograph (silica gel, gradient of 5%, 7.5%, 10% MeOH / CH2Cl2 + NH4OH) to give 18.49 g of the title compound as a racemic mixture.

Step D - Separation of enantiomers H The racemic title compound from step C is separated by preparative chiral chromatography (Chiralpack AD column, 5 cm x 50 cm, flow rate 100 mL / min., 20% PrOH / hexane + 0.2% diethylamine), to give 9.14 g of the isomer (+) and 9.30 g of the isomer (-). Physicochemical data for the (+) isomer: p.f. = 74.5 ° -77.5 ° C; Mass spectrum: MH + = 471.9; [a] = + 97.4 ° (8.48 mg / 2 mL of MeOH). Physicochemical data for the (-) isomer: p.f. = 82.9 ° - 84.5 ° C; Mass spectrum: MH + = 471.8; [a] = -97.4 ° (8.32 mg / 2mL MeOH).

EXAMPLE OF PREPARATION 4 Step A g (38.5 mmol) of ethyl ester of 4- (8-chloro-3-bromo-5,6-dihydro-11 H-benzo [5,6] cyclopehta [1,2-b] pyridine) are combined. 11-ylidene) -1-piperidino-1-carboxylic acid and 150 mL of concentrated H2SO at -5 ° C, then 3.89 g (38.5 mmoles) of KNO3 are added and stirred for 4 hours. The mixture is poured into 3 L of ice and made basic with 50% NaOH (aqueous). Extract with CH2Cl2, dry over MgSO4, then filter and concentrate in vacuo to a residue. Recrystallization of the residue from acetone gives 6.69 g of the product. 1 H NMR (CDCl 3, 200 MHz): 8.5 (s, 1 H); 7.75 (s, 1H); 7.6 (s, 1 H); 7.35 (s, 1 H); 4.15 (q, 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).

Step B 6.69 g (13.1 mmol) of the product from step A and 100 mL of 85% EtOH / water are combined, 0.66 g (5.9 mmol) of C and 6.56 g (117.9 mmol) of Fe are added and the mixture is heated to 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 give 7.72 g of the product. Mass spectrum: MH + = 478.0 Step C Combine 7.70 g of the product from step B and 35 mL of HOAc, then add 45 mL of a Br2 solution in HOAc and stir the mixture at room temperature overnight. 300 mL of 1 N NaOH (aqueous) is added, then 75 mL of 50% NaOH (aqueous) and extracted with EtOAc. The extract is dried over MgSO 4 and concentrated in vacuo to a residue. Chromatography of the residue (silica gel, 20% -30% EtOAc / hexane) gave 3.47 g of the product (along with another 1.28 g of a partially purified product). Mass spectrum: MH + = 555.9. 1 H NMR (CDCl 3, 300 MHz): 8.5 (s, 1 H); 7.5 (s, 1 H); 7.15 (s, 1 H); 4.5 (s, 2H); 4.15 (m, 3H); 3.8 (br s, 2H); 3.4-3.1 (m, 4H); 9-2.75 (m, 1 H); 2.7-2.5 (m, 2H); 2.4-2.2 (m, 2H); 1.25 (m, 3H).

Step D Combine 0.557 g (5.4 mmol) of t-butylnitrite and 3 mL of DMF, and the mixture is heated 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 reheated to 60 ° -70 ° C for 0.5 hours. It is cooled to room temperature and the mixture is poured into 150 mL of water. Extract with CH2Cl2, dry over MgSO4 and concentrate in vacuo to a residue. Chromatography of the residue (silica gel, 10% -20% EtOAc / hexane) gave 0.74 g of the product. Mass Spectrum: MH + = 541.0 1 H NMR (CDCls, 200 MHz): 8.52 (s, 1 H); 7.5 (d, 2H); 7.2 (s, 1 H); 4.15 (q, 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).

Step E Combine 0.70 g (1.4 mmol) of the product from step D and 8 mL of concentrated HCl (aqueous) and the mixture is refluxed overnight. 30 mL of 1 N NaOH (aqueous) is added, then 5 mL of 50% NaOH (aqueous) and extracted with CH2Cl2. The extract is dried over MgSO4 and concentrated in vacuo to give 0.59 g of the title compound. Mass spectrum: M + = 468.7, p.f. = 123.9 ° -124.2 ° C.

EXAMPLE OF PREPARATION 5 [racemic, as well as isomers (+) and (-)] Step A A solution of 8.1 g of the title compound is prepared from Preparation Example 4 in toluene, and 17.3 mL of a 1M DIBAL solution in toluene are added. The mixture is heated to reflux and slowly (drip) another 21 mL of 1 M DIBAL solution / toluene is added over a period of 40 minutes. The reaction mixture is cooled to about 0 ° C and 700 mL of 1 M HCl (aqueous) is added. The organic phase is separated and discarded. The aqueous phase is washed with CH2Cl, the extract is discarded and made basic by adding 50% NaOH (aqueous). Extract with CH2Cl2, dry the extract over MgSO4 and concentrate in vacuo to give 7.30 g of the title compound, which is a racemic mixture of enantiomers.

Step B - Separation of enantiomers The racemic title compound from step A is separated by preparative chiral chromatography (Chiralpack AD column, 5 cm x 50 cm, using 20% PrOH / hexane + 0.2% diethylamine), to give the (+) isomer and the isomer (-) of the title compound. Physicochemical data for the (+) isomer: p.f. = 148.8 ° C; Mass spectrum: MH + = 469; [a] = + 65.6 ° (mg / 2mL MeOH). Physicochemical data for the (-) isomer: p.f. = 112 ° C; Mass spectrum: MH + = 469; [a] = - 65.2 ° (mg / 2mL MeOH).

EXAMPLE OF PREPARATION 6 [racemic, as well as isomers (+) and (-)] Step A Combine 40.0 g (0.124 mol) of the starting ketone and 200 mL of H2SO4 and cool to 0 ° C. Slowly add 13.78 g (0.136 mol) of KNO3 over a period of 1.5 hours, then warm to room temperature and stir overnight. Treatment of the reaction using substantially the same procedure as that described for preparation example 1, step A. Chromatography (silica gel, 20%, 30%, 40%, 50% EtOAc / hexane, then 100% EtOAc) gave 28 g of the 9-nitro product, together with a smaller amount of the 7-nitro product and 19 g of the 7-nitro and 9-nitro compounds.

Step 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 than that described for preparation example 1, step C, to give 24 g of the product.

Step C Combine 13 g (38.5 mmol) of the product from step B, 140 mL of HOAc and slowly add a solution of 2.95 mL (57.8 mmol) of Br2 in 10 mL of HOAc for a period of 20 minutes. The reaction mixture is stirred at room temperature, then concentrated in vacuo to a residue. CH2CL2 and water are added, then adjusted to pH = 8.9 with 50% NaOH (aqueous). The organic phase is washed with water, then brine and dried over Na2SO4. Concentrate in vacuo to give 11.3 g of the product.

Step D Cool 100 mL of concentrated (aqueous) HCl at 0 ° C, then add 5.61 g (81.4 mmol) of NaNO2 and stir for 10 minutes. 11.3 g (27.1 mmoles) of the product from step C are added slowly (in portions) and the mixture is stirred at 0-3 ° C for 2.25 hours. Slowly add (dropwise) 180 mL of 50% H3PO2 (aqueous) and allow the mixture to stand at 0 ° C overnight. Slowly add (dropwise) 150 mL of 50% NaOH for 30 minutes, to adjust to pH = 9, then extract with CH2Cl2. The extract is washed with water, then brine and dried over Na2SO4. Concentrate in vacuo to a residue and chromatograph (silica gel, 2% EtOAc / CH 2 Cl 2) to give 8.6 g of the product.

Step E 8.6 g (21.4 mmol) of the product from step D and 300 mL of MeOH are combined and cooled to 0-2 ° C. 1.21 g (32.1 mmoles) of NaBH 4 is added and stirred at ~ 0 ° C for 1 hour. Another 0.121 g (3.21 mmol) of NaBH are added, stirred for 2 hours at 0 ° C, and then left to stand overnight at 0 ° C. Concentrate in vacuo to a residue and then remove the residue between CH2Cl2 and water. The organic phase is separated and concentrated under vacuum (50 ° C) to give 8.2 g of the product.

Step F 8.2 g (20.3 mmol) of the product from step E and 160 mL of CH2Cl2 are combined, cooled to 0 ° C, then 14.8 mL (203 mmol) of SOCI are added slowly (dropwise) over a period of 30 minutes. The mixture is warmed to room temperature and stirred for 4.5 hours, then concentrated in vacuo to a residue, CH 2 Cl 2 is added and washed with 1 N NaOH (aqueous) then brine and dried over Na 2 SO 4. Concentrate in vacuo to a residue, then add dry THF and 8.7 g (101 mmol) of piperazine and stir at room temperature overnight. Concentrate in vacuo to a residue, add CH2CI2 and wash with 0.25 N NaOH (aqueous), water and then brine. Dry over Na2SO4 and concentrate in vacuo to give 9.46 g of the crude product. Chromatography (silica gel, 5% MeOH / CH 2 Cl 2 + NH 3) gave 3.59 g of the title compound as a racemate. 1 H NMR (CDCl 3, 200 MHz): 8.43 (d, 1 H); 7.55 (d, 1 H); 7.45 (d, 1 H); 7.11 (d, 1H); 5.31 (s, 1 H); 4.86-4.65 (m, 1 H); 3.57-32.40 (m, 1 H); 2.98-2.55 (m, 6H); 2.45-2.20 (m, 5H).

Step G - Separation of enantiomers The racemic title compound from step F (5.7 g) is subjected to chromatography as described for Preparation Example 3, step D, using 30% PrOH / hexane + 0.2% diethylamine, to give 2.88 g of the R-isomer - (+) and 2.77 g of the S - (-) isomer of the title compound. Physicochemical data for the R - (+) isomer: Mass spectrum: MH + = 470; [a] = + 12.1 ° (10.9 mg / 2 mL of MeOH). Physicochemical data for the S - (-) isomer: Mass spectrum: MH + = 470; [a] = -13.2 ° (11.51 mg / 2 mL of MeOH).

EXAMPLE OF PREPARATION 7 [racemic, as well as isomers (+) and (-)] Step A Combine 13 g (33.3 mmol) of the title compound of Preparation Example 1, Step D, and 300 mL of toluene at 20 ° C, then add 32.5 mL (32.5 mmol) of a 1M DIBAL solution in toluene. The mixture is refluxed for 1 hour, cooled to 20 ° C, another 32.5 mL of 1M DIBAL solution is added and it is heated at reflux for 1 hour. The mixture is cooled to 20 ° C and poured into a mixture of 400 g of ice, 500 mL of EtOAc and 300 mL of 10% NaOH (aqueous). The aqueous layer is extracted with CH2Cl2 (3 x 200 mL), the organic layers are dried over MgSO4, then concentrated in vacuo to a residue. Chromatography (silica gel, 12% MeOH / CH 2 Cl 2 + 4% NH 4 OH) gave 10.4 g of the title compound as a racemate. Mass spectrum: MH + = 469 (FAB). 1 H Partial NMR (CDCl 3, 400 MHz): 8.38 (s, 1 H); 7.57 8s, 1 H); 7.27 (d, 1 H); 7.06 (d, 1 H); 3.95 (d, 1 H).

Step B - Separation of enantiomers The racemic title compound from step A is separated by preparative chiral chromatography (Chiralpack AD column, 5 cm x 50 cm, using 5% iPrOH / hexane + 0.2% diethylamine), to give the (+) isomer and the isomer ( -) of the title compound. Physicochemical data for the (+) isomer: Mass spectrum: MH + = 470.9 (FAB); [α] 0 + 43.5 ° (C = 0.402, EtOH); 1 H Partial NMR (CDCl 3, 400 MHz): 8.38 (s, 1 H); 7.57 (s, 1 H); 7.27 (d, 1 H); 7.05 (d, 1 H); 3.95 (d, 1 H). Physicochemical data for the (-) isomer: Mass spectrum: MH + = 470.9 (FAB); [a] = -41.8 ° (c = 0.328 EtOH); 1 H Partial NMR (CDCl 3, 400 MHz): 8.38 (s, 1 H); 7.57 (s, 1 H); 7.27 (d, 1 H); 7.05 (d, 1 H); 3.95 (d, 1 H).

EXAMPLE OF PREPARATION 8 [racemic, as well as R - (+) and S - (-) isomers] It is treated with ethyl ester of 4- (8-chloro-3-bromo-5,6-dihydro-11 H -benzo [5,6] cyclohepta [1,2- b] pyridin-11-liden) - 1-piperidin-1 -carboxylic substantially by means of the same procedure as that described in Preparation Example 3, steps AD, to give as the product of step 8 C, the compound of the radicle title, and as the products of step D the R - (+) isomer and the S - (-) isomer of the title compound. Physicochemical data for the R - (+) isomer: 13C NMR (CDCl 3): 155.8 (C); 146.4 (CH); 140.5 (CH); 140.2 (C); 136.2 (C); 135.3 (C); 133.4 (C); 132.0 (CH); 129.9 (CH); 125.6 (CH); 119.3 (C); 79.1 (CH); 52.3 (CH2); 52.3 (CH); 45.6 (CH2); 45.6 (CH2); 30.0 (CH2); 29.8 (CH2). [a] = + 25.8 ° (8.46 mg / 2 mL of MeOH). Physicochemical data for the S - (-) isomer: 13C NMR (CDCl 3): 155.9 (C); 146.4 (CH); 140.5 (CH); 140.2 (C); 136.2 (C); 135.3 (C); 133.3 (C); 132.0 (CH); 129.9 (CH); 125.5 (CH); 119.2 (C); 79.1 (CH); 52.5 (CH2); 52.5 (CH); 45.7 (CH2); 45.7 (CH2); 30.0 (CH2); 29.8 (CH2). [a] = -27.9 ° (8.90 mg / 2mL of MeOH).

EXAMPLE OF PREPARATION 9 9 Step A 9.90 g (18.9 mmol) of the product of Preparation Example 4, Step B, are dissolved in 150 mL of CH2Cl2 and 200 mL of CH3CN and heated to 60 ° C. 2.77 g (20.8 mmol) of N-chlorosuccinimide are added and the mixture is refluxed for 3 hours, monitoring the reaction by CCD (30% EtOAc / H2O). An additional 2.35 g (10.4 mmoles) of N-chlorosuccinimide are added and refluxed for an additional 45 minutes. The reaction mixture is cooled to. The ambient temperature is extracted with 1 N NaOH and CH2Cl2. The CH2Cl2 layer is dried over MgSO4, filtered and purified by flash chromatography (1200 mL of normal phase silica gel, eluting with 30% EtOAc / H20) to obtain 6.24 g of the desired product. P.f. 193-195.4 ° C.

Step B To 160 mL of concentrated HCl at -10 ° C is added 2.07 g (30.1 mmol) of NaN02 and stirred for 10 minutes. 5.18 g (10.1 mmol) of the product from step A are added and the reaction mixture is heated from -10 ° C to 0 ° C for 2 hours. The reaction is cooled to -10 ° C, 100 mL of H3PO2 is added and it is left to stand overnight. To extract the reaction mixture, it is poured onto crushed ice and made basic with 50% NaOH / CH2Cl2. The organic layer is dried over MgSO, filtered and concentrated to dryness. Purification is done by flash chromatography (600 mL of normal phase silica gel, eluting with 20% EtOAc / hexane) to obtain 3.98 g of the product. Mass spectrum: MH + = 497.2.

Step C 3.9 g of the product from step B are dissolved in 100 mL of concentrated HCl and refluxed overnight. The mixture is cooled, made basic with 50% w / w NaOH and the resulting mixture is extracted with CH 2 Cl 2. The CH2Cl2 layer is dried over MgSO4, the solvent is evaporated and dried under vacuum to obtain 3.09 g of the desired product. Mass spectrum: MH + = 424.9.

Step D Using a procedure similar to that described in Preparation Example 5, 1.73 g of the desired product is obtained, m.p. 169.6-170.1 ° C; [a] = + 48.2 ° (c = 1, MeOH).

EXAMPLE OF PREPARATION 10 Step A Combine 1.33 g of the (+) enantiomer of the compound of Preparation Example 5, step B, in anhydrous DMF with 1.37 g of 1-N-t-butoxy-carbonylpiperidinyl-4-acetic acid, and with DEC, HOBT and N-methylmorpholine. The mixture is stirred at room temperature overnight. Concentrate in vacuo to remove the DMF and add 50 mL of saturated NaHCO3 (aqueous). Extract with CH2C12 (2 x 250 mL), extract the extracts with 50 mL of brine and dry over MgSO4. Concentrate in vacuo to a residue and chromatograph (silica gel, 2% CH3OH / CH2Cl2 + 10% NH4OH) to give 2.78 g of the product. Mass spectrum: MH + = 694.0 (FAB); [a] = + 34.1 ° (5.45 mg / 2 mL, MeOH).

Step B 2.78 g of the product from step A and CH2Cl2 are combined, then cooled to 0 ° C and TFA is added. The mixture is stirred for 3 hours at 0 ° C, then 1N NaOH (aqueous) is added, followed by 50% NaOH (aqueous). Extract with CH2Cl2, dry over MgSO4 and concentrate in vacuo to give 1.72 g of the product. P.f. = 104.1 ° C; Mass spectrum: MH + = 594; [] = + 53.4 ° (1 1.42 mg / 2 mL, CH3OH).

PREPARATION EXAMPLE 11 (+) - 1 - (Aminoacetyl) -4- (3-dibromo-8-chloro-6,11-dihydro-5H-benzor5,61cycloheptaH, 2-b1pyridin-11 -Dipelinein Step 1: (+) - 1.1-D-methylethyl-r4- (3,10-dibromo-8-chloro-6,11-dihydro-5H-benzor-5,61-cyclohepta [1,2-b-pyridin-11-yl] -1-piperidinin-2-oxoetill-carbamate The product of Preparation Example 5 (+ isomer) (0.4 g, 0.85 mmol) was dissolved in DMF (10 mL) and then cooled to ~ 4 ° C. Then BOC-glycine (0.19 g, 1.1 mmol) was added, followed by DEC (0.2 g, 1.1 mmol), HOBT (0.15 g, 1.1 mmol) and 4-methylmorpholine (0.11 g, 0.12 μL, 1.1 mmol). The reaction was stirred at room temperature overnight, then concentrated in vacuo to a residue and separated between CH 2 Cl 2 and saturated NaHC 3 (aqueous). The aqueous phase was further extracted with CH2Cl2, the combined CH2Cl2 fractions were dried over MgSO4 and concentrated in vacuo to give a residue which was subjected to column chromatography on silica gel using 5% (saturated NH3 CH3OH) / CH2Cl2 as the eluent to give the title compound as a white solid: 0.52 g, 99% yield, mp = 95-96 ° C, MH + = 628.

Step 2 The product from step 1 (2.65 g, 4.2 mmol) was dissolved in CH2Cl2 (20 mL) and cooled to 0 ° C. Then trifluoroacetic acid (10 mL) was added. The reaction mixture was stirred at room temperature for 4 hours, then poured onto ice and the pH was adjusted to 10 using 50% aqueous (w / v) NaOH. The reaction mixture was extracted with CH2Cl2, the combined CH2Cl2 extracts were washed with H2O and brine, and dried over Na2SO4. The solvents were removed by rotary evaporation to give the title compound as a white solid: 2.18 g, 98% yield, m.p. = 150-152 ° C, MH + = 528.

EXAMPLE OF PREPARATION 12 (+) - 1 - (3-Amino-1-oxopropyl) -4- (3-dibromo-8-chloro-6,11-dihydro-5H-benzor-5,61-cycloheptaph1,2-pyridine-11 -iOpiperidine Step 1: (+) - 1.1-D-methylethyl-3- [4- (3,10-dibromo-8-chloro-6,11-dihydro-5H-benzoyl-5,61-cycloheptyl, 2-b-1-pyridin-11-yl ) -1-pperidinyl-1-oxopropylcarbamate The title compound was prepared following essentially the same procedure as that described in Preparation Example 11, Step 1, except that BOC-β-alanine was used in place of BOC-glycine to obtain a white solid. Yield = 99%, MH + = 642.

Step 2 The title compound was prepared following essentially the same procedure as that described in Preparation Example 11, Step 2, to obtain a white solid. Yield = 100%, m.p. = 136-137 ° C, MH + = 642.

EXAMPLE OF PREPARATION 13 (+) - 4-.3.10-Dibromo-8-chloro-6.11-dihydro-5H-benzo.5.61cyclohepta-p .2- blpiridin-11-yl) -1-r4-amino .-1 -oxofautillpiperidina Step 1: (+) - 1,1-dimethylethyl [4-r4- (3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo [5,61-cycloheptyl, 2-b-pyridin-11-yl] -1-piperidinyl-2-oxobutylcarboxamide The title compound was prepared following essentially the same procedure as that described in Preparation Example 11, Step 1, except that BOC-a-amino-butyric acid was used in place of BOC-glycine to obtain a white solid. Yield = 79%, m.p. = 102-103 ° C, MH + = 781.

Step 2 The title compound was prepared following essentially the same procedure as that described for preparation example 11, step 2, to obtain a white solid. Yield = 94%, m.p. = 114-115 ° C, MH + = 681.

EXAMPLE OF PREPARATION 14 f +) - 1-fAminoacetyl) -4-r2-r4- (3,10-dibromo-8-chloro-6,11-dihydro-5H-benzof5,61cycloheptyl, 2-b) pyridine-11 -yl) -1-piperidinin-2-oxoetipiperidine Step 1: (+) - 1,1-Dimethylol [2-r4- [2- [4- (3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo [5,6-chloro-hepta] [1,2-blpyridin-11-yl) -1-piperidinin-2-oxoetin-1-piperidinyl-2-oxoetylcarbamate The title compound was prepared following essentially the same procedure as that described for Preparation Example 11, Step 1, except that the compound of Preparation Example 10 (+ isomer) was used in place of the compound of Preparation Example 5 to obtain a white solid. Yield = 82%, m.p. = 98-99 ° C, MH + = 753.

Step 2 The title compound was prepared following essentially the same procedure as that described in Preparation Example 11, Step 2, to obtain a white solid. Yield = 89%, m.p. = 130-131 ° C, MH + = 653.

PREPARATION EXAMPLE 15 (+) - 1-f3-Arnino-1-oxopropyl) -4-r2-r4- (3,10-dibromo-8-chloro-6,11-dihydro-5H-benzor5,61c5clo-hepta. 1, 2-bl | pirin-11 -yl) -1-piperidinin-2-oxoetinpiperidine Step 1: (+) - 1.1-D-methyletiir3-r4-r2-r4- (3,10-dibromo-8-chloro-6,11-dihydro-5H-benzo [5,61cycloheptaH, 2-b1pyridin- 11 -yl) -1-p -peridinin-2-oxoethyl-1-piperidinip-3-oxopropylcarbamate The title compound was prepared following essentially the same procedure as that described for Preparation Example 12, Step 1, except that the compound of Preparation Example 10 was used in place of the compound of Preparation Example 5 to obtain a white solid.

Yield = 84%, m.p. = 87-88 ° C, MH + = 767.

Step 2 The title compound was prepared following essentially the same procedure as that described for preparation example 11, step 2, to obtain a white solid. Yield = 84%, m.p. = 120-121 ° C, MH + = 667.

EXAMPLE OF PREPARATION 16 (+) - 1- (4-Amino-1-oxobutyl) -4-r2-r4- (3,10-dibromo-8-chloro-6,11-dihydro-5H-benzoid. -heptaH, 2-blpirdin-11 -yl) -1-piperidin-2-oxoetinpiperidine Step 1: f +) - 1.1-Dimethylethyl-4-r4-y2-r4- (3.10-dibromo-8-chloro-6,11-dihydro-5H-benzo [5,6-cycloheptap-2-blpyridin-11-yl] -1 -piperidin-2-oxoetin-1-piperidinyl-4-oxobutincarbamate The title compound was prepared following essentially the same procedure as that described for Preparation Example 13, Step 1, except that the compound of Preparation Example 10 was used in place of the compound of Preparation Example 5 to obtain a white solid. Yield = 79%, m.p. = 102-103 ° C, MH + = 782.

Step 2 The title compound was prepared following essentially the same procedure as that described for preparation example 11, step 2, to obtain a white solid. Yield = 94%, m.p. = 114-115 ° C, MH + = 681.

EXAMPLE OF PREPARATION 17 Dissolve 2 g (12.7 mmol) of methyl 2-oxo-1-pyrrolidino acetate in 20 mL of EtOH and then add 20 mL of 1M LiOH. The reaction mixture is stirred at room temperature for 16 hours. The solvents are evaporated, the resulting material is dissolved in water and the pH is adjusted to ~4. The reaction mixture is concentrated to give the product. Mass spectrum: MH + = 144.

EXAMPLE 1 (+) - 4-.3,10-Dibromo-8-chloro-6.11 -dihydro-5H-benzof5,61cycle epta.1.2- blpiridin-11 -yl) -1- (2-oxo-1-pyrrolidineDacetillpiperidine The compound of Preparation Example 15 (0.15 g, 0.32 mmol) was dissolved in DMF (5 mL) and then cooled to about 4 ° C. The compound of Preparation Example 17 (0.06 g, 0.4 mmol) was then added, followed by DEC (0.08 g, 0.4 mmol), HOBT (0.6 g, 0.4 mmol) and 4-methylmorpholine (0.04 g, 50 μL, 0.4 mmol). , and the reaction was then stirred at room temperature overnight. The reaction mixture was concentrated in vacuo to a residue which was separated between CH 2 Cl 2 and saturated NaHCO 3 (aqueous). The aqueous phase was further extracted with CH 2 Cl 2, the combined CH 2 Cl 2 fractions were dried over MgSO 4 and concentrated in vacuo to give a residue which was chromatographed on a silica gel column using (5% NH 3 CH 3 OH saturated) / CH 2 Cl 2 as eluent for give the title compound as a white solid: 0.11 g, 61% yield, mp = 118-119 ° C, MH + = 596.

EXAMPLE 2 (+ -4- (3,10-Dibromo-8-chloro-6.11-dihydro-5H-benzoyl-5,6] cyso-hepta-1, 2-b] pyridin-11-iQ-1 -f 1 - oxo-3- (2-oxo-1-pyrrolidiniDpropippiperidin The title compound of Preparation Example 12 (0.4 g, 0.74 mmol) was dissolved in CH2Cl2 (10 mL) and then 4-bromobutyryl chloride (0.2 g, 0.13 mL, 1.11 mmol) and Et3N (0.164 g, 0.23 g. mL, 1.62 mmol). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was separated between saturated NaHCOs and CH2Cl2. The aqueous phase was extracted with CH2Cl2, the combined CH2Cl2 extracts were dried over MgSO4 and the solvent was removed by rotary evaporation. The resulting product was dissolved in THF (10 mL), cooled to -10 ° C, NaH (0.09 g, 3.79 mmol) was added and the reaction mixture was stirred for 16 hours, allowing the temperature to rise to the temperature ambient. The reaction mixture was then separated between saturated aHCOs and EtOAc. The organic phase was dried over MgSO and purified by flash chromatography on silica gel, eluting with 3% CH3OH (saturated with NH3) / CH2Cl2 to give the title compound as a white solid (0.07 g), m.p. = 128-129 ° C, MH + = 610.

EXAMPLE 3 (+) - 4-α3.10-D-bromo-8-chloro-6,11-dihydro-5 H -benzor 5,61 cycloheptap.2-b.pyridin-11-yl) -1-1 -oxo- 4- (2-oxo-1-pyrrolidiniDbutinpiperidine The title compound is prepared from the product of Preparation Example 13, following essentially the same procedure as that described for Example 2, to obtain a white solid. P.f. = 127-128 ° C, MH = 624.

EXAMPLE 4 (+) - 4- (3,10-Dibromo-8-chloro-6,11-dihydro-5H-benzor-5,61-cycloheptap.2-blpyridin-11-yl) -1-r (2-oxo-1 - piperidinipacetillpiperidina The title compound is prepared from the product of Preparation Example 11, following essentially the same procedure as that described for Example 2, but 4-bromovaleryl chloride is used in place of 4-bromobutyryl chloride to obtain a white solid . Yield = 50%, m.p. = 138-139 ° C, MH = 610.

EXAMPLE 5 (+) - 4- (3,10-Dibromo-8-chloro-β, 11-dihydro-5H-benzor 5,61-cycloheptap, 2-blpiridin-11 -ID-1-1 -oxo-3- ( 2-oxo-1-piperidinippropinpiperidine The title compound was prepared from the product of Preparation Example 12, following essentially the same procedure as that described for Example 2, but 4-bromovaleryl chloride was used in place of 4-bromobutyryl chloride to obtain a white solid. Yield = 50%, m.p. = 138-139 ° C, MH = 610.

EXAMPLE 6 (+) - 4-.3.1 QD-bromo-8-chloro-6,11-dihydro-5H-benzo.5,61-cyclohepta-1, 2- b.pyridin-11-yl) -1 -ri - oxo-4- (2-oxo-1-piperidiniDbutinpiperidine The title compound is prepared from the product of Preparation Example 13, following essentially the same procedure as that described for Example 2, except that 4-bromovaleryl chloride is used in place of 4-bromobutyryl chloride to obtain a white solid. Yield = 81%, m.p. = 101-102 ° C, MH = 638.

EXAMPLE 7 (+) - 4- (3,10-Dibromo-8-chloro-6,11-dihydro-5H-benzo-5,61-cyclohepta-H, 2-blpyridin-11-yl) -1 -r ( 2-oxo-1-imidazolidinyl) acetylpipepdine The product of Preparation Example 11 (2.08 g, 3.9 mmol) was dissolved in CH2Cl2 (20 mL) and 2-bromoethyl isocyanate (0.8 g, 0.5 mL, 7.9 mmol) was added. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was separated between NaHCOs and CH2Cl2. The aqueous phase was extracted with CH2Cl2, the combined CH2Cl2 extracts were dried over MgSO4 and the solvent was removed by rotary evaporation. The resulting product was dissolved in THF (20 mL), cooled to -10 ° C, NaH (0.46 g, 19.5 mmol) was added and the reaction mixture was stirred for 16 hours, allowing the temperature to reach room temperature. . The reaction mixture was then separated between saturated NaHCO3 and EtOAc. The organic phase was dried over MgSO 4 and purified by flash chromatography on silica gel, eluting with 5% CH 3 OH (saturated with NH 3) / CH 2 Cl 2 to give the title compound as a white solid: 1.95 g, yield = 80% , pf = 167-168 ° C, MH + = 597.

EXAMPLE 8, +) - 4- (3,10-Dibromo-8-chloro-6,11-dihydro-5H-benzo.5,61cyclohepta.1.2-b1pyridin-11-yl) -1.-r3-f2 -oxoimidazolidinyl) -1-oxopropinpiperidine The title compound is prepared from the product of preparation example 12, following essentially the same procedure as that described in example 7 to obtain a white solid. Yield = 45%, m.p. = 202-203 ° C, MH + = 611.

EXAMPLE 9 ((+) 4- (3.10-Dibronno-8-chloro-6.11-dihydro-5H-benzor5.61cycloheptari.2- blpiridin-11-IQ-1 -oxo-4- (2-oxo-1- midazolidinyl) butinpiperidine The title compound is prepared from the product of Preparation Example 13, following essentially the same procedure as that described in Example 7 to obtain a white solid. Yield = 52%, m.p. = 120-123 ° C, MH + = 625.

EXAMPLE 10 (+) - 4- (3,10-Dibromo-8-chloro-6,11-dihydro-5H-benzor5.61cycloheptan.2-b.pyridin-11 -ID-1-y (exahydro-2-oxo -1-pyrimidine Oacetypyperidine The title compound is prepared from the product of Preparation Example 11, following essentially the same procedure as that described for Example 7, substituting 2-bromoethyl isocyanate with 3-chloropropyl isocyanate to obtain a white solid. Yield = 56%, m.p. = 155-156 ° C, MH + = 611.

EXAMPLE 11 (+) - 4- (3,1 Q-Dibromo-8-chloro-6,11-dihydro-5 H -benzof 5,61 cycloheptari, 2-blpiridin-11 -5D-1 -H -oxo-3- (hexalhido -2-oxo-1-pyrimidinyl) - oxopropinpiperidine H The title compound is prepared from the product of preparation example 12, following essentially the same procedure as described for Example 10 to obtain a white solid. Yield = 40%, m.p. = 135-136 ° C, MH + = 625.

EXAMPLE 12 (+) - 4- (3,10-D-bromo-8-chloro-6,11-dihydro-5H-benzor5,6.cycloheptap, 2-blpyridin-11-yl) -1-f4- (hexahydrogen -2-oxo-1-pyrimidlniOoxobutiripiperidirta The title compound is prepared from the product of Preparation Example 13, following essentially the same procedure as that described for Example 10 to obtain a white solid. Yield = 63%, m.p. = 157-158 ° C, MH + = 639. The compounds of the following structure are prepared using the starting materials described above and the appropriate procedure: FPT IC50 (in vitro enzyme inhibition assay of farnesyl protein transferase) IC50 of COS cells (cell-based assay) are determined., GGPT IC50 (in vitro enzyme inhibition test of geranylgeranyl-protein transferase) Test of Cell Matter and antitumor activity (antitumor studies in vivo) by the test procedures described in WO 95/10516. The test protocol for the evaluation used to determine the inhibition of the growth of tumor cells in humans, the Soft Agar Test, is as follows: The independent growth of anchoring is a characteristic of Isa tumorigenic cell lines. Human tumor cells are suspended in growth medium containing 0.3% agarose and an indicated concentration of a farnesyl transferase inhibitor. The solution is placed on the growth medium solidified 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 5% C02 to allow the growth of colonies.

After incubation, the colonies are stained by placing on the agar a solution of MTT ((3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide, thiazolyl blue) (1 mg / mL in PBS). The mutation state of Ras was determined by ELISA (Oncogene Science). The colonies are counted and the IC50s are determined. The IC50 FPT values determined for the compounds of this invention ranged from 0.0014 to 0.085 μM. The IC50 values for the inhibition of ras processing in COS cells varied from O.010 to 0.16 μM for the compounds of this invention. The results of the soft agar test using NIH 3T3 H ras activated by tumor cell line ranged from 0.145 to > 0.500 for the compounds of the invention.

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, lozenges and suppositories. The powders and tablets may comprise about 5 to about 70% active ingredient. Suitable solid carriers are known in the art, for example, magnesium carbonate, magnesium stearate, talc, sugar, lactose. Tablets, powders, pills and capsules can be used as solid dosage forms suitable for oral administration. To prepare 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 by stirring. The molten homogeneous mixture is then poured into molds of suitable size, allowed to cool and then solidified. 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 powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas. Also included are solid preparation forms which are designed to be converted, just before use, into liquid form preparations for oral or parenteral administration. Said liquid forms include solutions, suspensions and emulsions. The compounds of the invention can also be delivered transdermally. The transdermal compositions may take the form of creams, lotions, aerosols and / or emulsions, and may be included in a transdermal patch type matrix or receptacle, which 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 suitable quantities of the active component, for example, an effective amount to achieve the desired purpose. The amount of active compound in a unit dose of preparation can be varied or adjusted from about 0.1 mg to 1000 mg, most preferably about 1 mg to 300 mg, according to the particular application. The current dose used may vary depending on the requirements of the patient and the severity of the condition being treated. The determination of the appropriate dosage for a particular situation is within the abilities of a person skilled in the art. In general, the treatment starts with smaller doses that are lower than the optimum dose of the compound. Subsequently, the dose is increased in small increments until the optimum effect is reached under the circumstances. For reasons of convenience, the total daily dose 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 physician, considering factors such as age, condition and size of the patient, as well as the severity of the symptoms that are being treated. A typical dosage regimen that is recommended is oral administration of 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 in this dosage regimen. The following are examples of pharmaceutical dosage forms containing a compound of the invention. The scope of the invention in this aspect of pharmaceutical composition should not be limited by the examples provided.

Examples of pharmaceutical dosage form EXAMPLE A Tablets Method of manufacture Ingredients 1 and 2 are mixed in a suitable mixer for 10-15 minutes. The mixture is granulated with ingredient 3. The wet granules are milled through a coarse screen (e.g., 0.63 cm) if necessary. The wet granules are dried. Dry granules are sieved if necessary and mixed with ingredient 4 and mixed for 10-15 minutes. Ingredient 5 is added and mixed for 1-3 minutes. The mixture is compressed to a suitable size and weighed on a suitable tablet machine.

EXAMPLE B Capsules Method of manufacture Ingredients 1, 2 and 3 are mixed in a suitable mixer for 10-15 minutes. Ingredient 4 is added and mixed for 1-3 minutes. The mixture is filled into suitable two-piece hard gelatin capsules in a suitable encapsulating machine. Although the present invention has been described in conjunction with the specific embodiments described above, many alternatives, modifications and variations thereof will be apparent to those skilled in the art. It is intended that all such alternatives, modifications and variations fall within the spirit and scope of the present invention.

Claims (11)

NOVELTY OF THE INVENTION CLAIMS

1. - A compound represented by the structural formula: or an N-oxide thereof, or a pharmaceutically acceptable salt or solvate thereof, wherein: R1 and R2 are independently selected from halogen; R1 and R3 are independently selected from the group consisting of H and halogen, as long as at least R1 and R3 are H; W is N, CH or C when the double bond is present at the C11 position; Z1 and Z2 are independently selected from the group consisting of = O and = S; n is 1-6 and neither is O or 1.

2. A compound according to claim 1, further characterized in that Z1 is = O.

3. A compound according to any of claims 1 or 2, further characterized in that R4 is

4. - A compound according to any of claims 1, 2 or 3, further characterized in that R is bromine and R2 is chlorine or bromine.

5. A compound according to any of claims 1, 2 or 3, further characterized in that R is bromine, R2 is chlorine or bromine, R1 is H and R3 is chlorine or bromine.

6. - A compound according to any of claims 1, 2 or 3, further characterized in that R is bromine, R2 is chlorine or bromine, R3 is H and R1 is chlorine or bromine. 1.

A compound according to claim 1, selected from the group consisting of

8. - A pharmaceutical composition for inhibiting abnormal cell growth, comprising an effective amount of a compound according to any of claims 1, 2, 3, 4, 5, 6, 7 or 8, in combination with a pharmaceutically acceptable carrier .

9. The use of a compound according to any of claims 1, 2, 3, 4, 5, 6, 7 or 8, in the preparation of a medicament for the treatment of tumor cells expressing an activated ras oncogene.

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

11. The use of a compound according to any of claims 1, 2, 3, 4, 5, 6, 7 or 8, in the preparation of a medicament for inhibiting farnesyl protein transferase. SUMMARY OF THE INVENTION Compounds of the formula (I) are described useful for inhibiting the function of Ras and therefore for inhibiting or treating the abnormal growth of cells, or a pharmaceutically acceptable salt or solvate thereof, wherein: R1 and R2 are halogen; R1 and R3 are H and halogen, as long as at least one is H; W is N, CH or C when the double bond is present at the C11 position; R4 is - (CH2) n-R5 or (II) R ° is R ° or (IV) Z1 and Z2 are independently selected from the group consisting of = O and JN / gfr * P99 / 1683F