FR2753196A1 - TRICYCLIC INDAZOLE DERIVATIVES, THEIR PREPARATION AND THEIR THERAPEUTIC APPLICATION - Google Patents

Abstract

The invention concerns compounds of general formula (I) in which: X represents one oxygen atom or one methene group, R1 represents one hydrogen atom, one halogen such as bromine, chlorine or fluorine, one C1-4 oxyalkylate group, or one hydroxyl or amino group; R2, R3, and R4, identical or different, represent either one hydrogen atom, or one C1-4 alkyl group, linear or branched, one phenyl group or one benzyl group, and R5 represents one hydrogen atom, one hydroxyl group, one amino group or one C1-4 alkyl group, in the form of enantiomer, of diastereoisomer, including racemic mixture as well as their additive salt to pharmaceutically acceptable acids. The compounds of formula I are antagonists of 5-HT3 and 5-HT4 serotoninergic receptors.

Description

The present invention relates to tricyclic indazole derivatives, their preparation and their therapeutic application.

dans laquelle
X représente un atome d'oxygène ou un groupe méthylène, Rl représente un atome d'hydrogène, un halogène tel que chlore ou fluor, un groupe C14 alcoxyle, ou un groupe hydroxyle ou amino, et
R2 et R3, identiques ou différents, représentent soit un atome d'hydrogène, soit un groupe C14 alkyle, linéaire ou ramifié, un groupe phényle ou un groupe benzyle.The compounds correspond to the general formula (I)

in which
X represents an oxygen atom or a methylene group, R1 represents a hydrogen atom, a halogen such as chlorine or fluorine, a C14 alkoxyl group, or a hydroxyl or amino group, and
R2 and R3, which may be identical or different, represent either a hydrogen atom or a linear or branched C14 alkyl group, a phenyl group or a benzyl group.

The compounds of general formula (I) contain one or more asymmetric carbon atoms. They can therefore exist in the form of enantiomers or diastereoisomers. These enantiomers, diastereoisomers, as well as their mixtures, including the racemic mixtures, form part of the invention.

The compounds of general formula (I) may be in free base form or addition salts with pharmaceutically acceptable acids, which are also part of the invention.

The compounds of the invention can be prepared by methods illustrated in the following diagrams.

The compounds of formula (I) can be prepared by reaction of a compound of formula (II), in which R 1 and R 2 are as defined in formula (I) and Y represents a halogen, in particular a chlorine atom, with a compound of formula (III), in which R 3 is defined as in formula (I), in a solvent such as chloroform or dichloromethane, in the presence of a basic salt such as potassium bicarbonate, according to the scheme 1. The reaction can be carried out at a temperature between -10 C and + 500C.

Diagram 1

The starting materials are directly commercially available, are known in the literature or can be synthesized by conventional methods known to those skilled in the art.

Thus the compound of formula (III) in which R 3 is a hydrogen atom is described in Arch. Pharmaz., 306 (12), pp. 934-942 (1973). The compounds of formula (III) in which
R3 is a methyl group may be prepared according to the process described in European Patent Application 0507650.

The compounds of formula (II), wherein Y is a chlorine atom and R1 and R2 are as defined in formula (I), may be prepared from a compound of formula (XVII) by activation of the carboxylic acid to acid chloride by means of thionyl chloride or oxalyl chloride according to methods known to those skilled in the art.

The compounds of formula (XVII) can be prepared according to the general method described in scheme 2, from derivatives of formula (IV), in which R1 and R2 are as defined in formula (I).

Figure 2

According to this scheme, the compounds of formula (IV) can be treated with boron trichloride and acetonitrile under the conditions described by T. Sugasawa et al., J. Am. Chem.

Soc., 100, pp. 1357, 1980, to give compounds of formula (V), wherein R1 and R2 are as defined in formula (I), followed by N-nitrosation with sodium nitrite in medium acid, under conditions known to those skilled in the art (Jerry March, Advanced Organic Chemistry, John Ed.
Wiley & Sons, 3rd ed., Pp 572), to give compounds of formula (VI). Compounds of formula (VI) can then be processed under Knoevenagel reaction conditions, known to those skilled in the art (Jerry March, Advanced Organic Chemistry, John Wiley & Sons ed., 3rd ed., Pp. 835-841) to give the compounds of formula (VII), wherein R1 and R2 are as defined in formula (I). The N-nitroso function of the compounds of formula (VII) can be reduced using zinc powder in the presence of acetic acid and immediately cyclized to give compounds of formula (VIII), wherein R1 and
R2 are as defined in formula (I). Finally, the compounds of formula (IX) can be prepared by oxidation of the compounds of formula (VIII), in the presence of ozone or by treatment with sodium periodate and osmium tetroxide and then be oxidized with nitrate of silver in the presence of potassium hydroxide to give compounds of formula (XVII), wherein R1 and R2 are as defined in formula (I).

Alternatively, the compounds of formula (XVII) in which R1 and R2 are as defined in formula (I) can be prepared according to the general method described in scheme 3.

Figure 3

According to this scheme, the compounds of formula (X) may be prepared from the derivatives of formula (IV) by reaction of 2-cyanofuran and boron trichloride under reaction conditions described by Sugasawa, referred to above. Compounds of formula (XII) can then be obtained by nitrosation of compounds of formula (X) followed by reduction of the resulting compounds of formula (XI) with zinc and subsequent cyclization of the intermediate. Finally, the compounds of formula (XII) are oxidized by treatment with potassium permanganate in a mixture of benzene and acetone (Chem Abs 55, p.16518, 1961) to give the compounds of formula (XVII), in which R1 and R2 are as defined in formula (I).

The compounds of formula (IV) may be prepared according to methods known to those skilled in the art.

For example, the compounds of formula (IV), wherein X is oxygen, R2 is hydrogen can be prepared by reacting a 2-aminophenol derivative with chloroacetyl chloride under phase transfer conditions (X. Huang et al., Synthesis, 10, pp. 851, 1984), followed by a reduction reaction using lithium aluminum hydride or a borane complex, generally in an ethereal solvent such as than tetrahydrofuran or diethyl ether.

Similarly, compounds of formula (IV), wherein X is oxygen, R2 is alkyl, phenyl or benzyl may be directly prepared by reaction of a 2-nitrophenol derivative with chloroacetone or 2-chloro or 2-bromoacetophenone in the presence of Raney nickel in ethanol (Melloni et al J. Het Chem 20, p.259, 1983).

The compounds of formula (I), in which R 2 represents a Cl 4 alkyl group, a phenyl or a benzyl group and X represents an oxygen atom, may be obtained in the form of enantiomers or diastereoisomers by preparation of compounds of formula (IV) optically active, according to scheme 4.

Figure 4

According to this process, an optically active amino-alcohol of formula (XIV) is activated, under the conditions described by Mitsunobu (Synthesis, 1981, pp 1-28), with 2-bromophenol or 2iodophenol of formula (XIII). The resulting ether (XV) is cyclized to the compound of formula (XVI), according to the conditions described by S. L. Buchwald et al., Angew. Chem. Int. Ed.

Engl. 1993, 34, pp 1348-1350 and deprotected according to methods known to those skilled in the art to give the compound of formula (IV).

Alternatively, the compounds of formula (I), in which R 2 represents a C 14 alkyl group, a phenyl or a benzyl, can be obtained in the form of enantiomers or diastereoisomers by performing a resolution with the aid of chiral salts of the compounds of formula (IV), (II) or (I).

The following examples illustrate the methods and techniques suitable for the preparation of this invention without, however, limiting the scope of the claim. Microanalyses and NMR and IR spectra confirm the structures of the compounds.

PREPARATION OF THE COMPOUNDS OF FORMULA (I)
Example 1: 2 - [[4- (5-methyl-1H-imidazol-4-yl) piperidin-1-yl] carbonyl] -7,8-dihydropyrazolo [1,5,4-de] [1 hydrochloride, 4] benzoxazine (2: 1) (compound no. 4 of the table, page 21).

a) Release from the base
1.15 ml of 5.3 N sodium methoxide is added to a solution of 0.7 g (0.0029 mol) of 4- (5-methyl-1H-imidazol-4-yl) piperidine dihydrochloride in 50 ml of methanol. . After stirring for 30 minutes, about 30 ml of solvent are evaporated off under reduced pressure. The precipitate formed is filtered and washed with chlororform and the filtrate containing the base is evaporated to dryness.

b) Coupling
A mixture of 0.28 g (0.0014 mol) of 7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid, 1 ml of thionyl chloride and a few drops of dimethylformamide in 30 ml of chloroform is refluxed for 2 hours. The solvent is evaporated under reduced pressure to obtain the acid chloride which will be used as it is subsequently. 4- (5-methyl-1H-imidazol-4-yl) piperidine (0.029 mol), obtained previously, is dissolved with 0.55 g (0.0039 mol) of potassium bicarbonate in 25 ml of chloroform. The mixture is cooled to 5 ° C. and a solution of the acid chloride (0.0014 mol) obtained previously in 30 ml of chloroform is added dropwise over 30 minutes and stirred at room temperature for 19 hours. The solvent is evaporated under reduced pressure and the residue is purified by chromatography on silica gel, eluting with a mixture of dichloromethane, methanol and ammonia (94: 6: 0.1).

0.32 g of product is obtained.

By adding a solution of hydrochloric ether in a mixture of ether and methanol to the base, the dihydrochloride is obtained.

M.p. = 154-156 ° C.

Example 2: 4-Fluoro-2 - [[4- (5-methyl-1H-imidazol-4-yl) piperidin-1-yl] carbonyl] -7,8-dihydropyrazolo [1,5,4-de] hydrochloride [1,4] Benzoxazine (1: 1) (Compound No. 5).

From 0.68 g (0.00306 mol) of 4-fluoro-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid and 1.46 g ( 0.00612 mol) of 4- (5-methyl-1H-imidazol-4-yl) piperidine, treated under the conditions of Example 1, after purification by chromatography on silica gel, eluting with a mixture of dichloromethane, methanol and ammonia (90: 10: 0.1), 0.984 g of product is obtained in free base form.

By adding to the product in the form of a base solution of hydrochloric ether in a mixture of ether and methanol, the dihydrochloride is obtained.

Melting point = 1900C.

Example 3: 2 - [[4- (5-Methyl-1H-imidazol-4-yl) piperidin-1-yl] carbonyl] -8-phenyl-7,8-dihydropyrazolo [2, 4-ethanedione] Ethanedioate [1,4] Benzoxazine (1: 1) (Compound No. 9).

From 0.16 g (0.00058 mol) of 8-phenyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid and 0.21 g ( 0.009 moles) of 4- (5-methyl-1H-imidazol-4-yl) piperidine, treated under the conditions of Example 1, after purification by chromatography on silica gel, eluting with a mixture of dichloromethane and methanol. and ammonia (92: 8: 0.1), 0.11 g of product in free base form is obtained.

By adding three equivalents of oxalic acid in methanol to the base product, oxalate is obtained.

Melting point = 140 ° C.

Example 4: 4-Fluoro-8-methyl-2 - [[4- (5-methyl-1H-imidazol-4-yl) piperidin-1-yl] carbonyl] -7,8-dihydropyrazolo [1,5-ethanedioate, 4-de] [1,4] benzoxazine (1: 1) (compound no. 10).

From 0.25 g (0.00106 mol) of 4-fluoro-8-methyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid and 0 39 g (0.0021 mol) of 4- (5-methyl-1H-imidazol-4-yl) piperidine, treated under the conditions of Example 1, after purififcation by chromatography on silica gel, eluting with a mixture of dichloromethane, methanol and ammonia (93: 7: 0.1), 0.18 g of product is obtained in free base form.

By adding three equivalents of oxalic acid in ethanol to the base product, oxalate is obtained.

Melting point = 243 ° C.

Example 5: (S) -8-Methyl-2 - [[4- (5-methyl-1H-imidazol-4-yl) piperidin-1-yl] carbonyl] -7,8-dihydropyrazolo [1,5-Ethanedioate , 4-de] [1,4] benzoxazine (1: 1) (compound no. 12).

From 0.13 g (0.00059 mol) of (S) -8-methyl-7,8dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid and 0.265 g (0.0009 mol) of 4- (5-methyl-1H-imidazol-4-yl) piperidine, treated under the conditions of Example 1, after purification by column of silica, eluting with a mixture of dichloromethane and methanol ( 97: 3), 0.13 g of product is obtained in free base form.

 [?] D20 = +53.3 (c = 0.01, methanol).

By adding three equivalents of oxalic acid in ethanol to the base product, oxalate is obtained.

Melting point = 2400C.

 [?] D20 = +52.5 (c = 0.01 dimethylsulfoxide).

Example 6: (R) -8-Methyl-2 - [[4- (5-methyl-1H-imidazol-4-yl) piperidin-1-yl] carbonyl] -7,8-dihydropyrazolo [1,5-Ethanedioate , 4-de] [1,4] benzoxazine (1: 2) (compound n013).

From 0.17 g (0.00077 mole) of (R) -8-methyl-7,8dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid and 0.325 g (0.0013 mole) 4- (5-methyl-1H-imidazol-4-yl) piperidine, treated under the conditions of Example 1, after purification by column of silica, eluting with a mixture of dichloromethane and methanol ( 97: 3), 0.15 g of product is obtained in free base form.

 [?] D20 = -38.20 (c = 0.01, methanol).

By adding three equivalents of oxalic acid in methanol to the base product, oxalate is obtained.

Melting point = 2360C.

 [?] D20 = -30.70 (c = 0.01 dimethylsulfoxide).

Example 7: [(4- (1H-Imidazol-4-yl) piperidin-1-yl] carbonyl] -7,8-dihydro-6H-pyrazolo [4,5-diis (2-E) -beta-2-enimidate 1-ij] quinoline (1: 2) (compound n01)
From 0.7 g (0.00346 mol) of 7,8-dihydro-6H pyrazolo [4,5,1-i] quinoline-2-carbocyclic acid and 1.5 g (0.0069 mol) of 4- (1H-Imidazol-4-yl) piperidine, treated under the conditions of Example 1, after purification by chromatography on silica gel, eluting with a mixture of dichloromethane, methanol and ammonia (95/5 / 0.1), 1 g of product is obtained in free base form.

By adding to the product as free base two equivalents of fumaric acid in ethanol, the fumarate is obtained.

Melting point = 202 ° C PREPARATION OF FORMULA INTERMEDIATES (XVII)
Example 8: 7,8-Dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid.

8.1. (3,4-Dihydro-2H-1,4-benzoxazin-5-yl) furan-2-yl methanone
To a solution of 15 g (0.11 mol) of 3,4-dihydro-2H-1,4-benzoxazine in 200 ml of 1,2-dichloroethane, maintained at a temperature between 0 and 50C and under nitrogen, is added slowly for 1 hour, a solution of 200 ml (0.2 mol) of boron trichloride (1 M in heptane). The mixture is refluxed for 1 h 30 min and then cooled before adding, for 20 minutes, a solution of 20 ml (0.22 mole) of 2-furonitrile in 20 ml of 1,2-dichloroethane. The reaction mixture is refluxed for 3 hours and then left overnight at room temperature.

In order to hydrolyze the imine intermediate formed, the reaction medium is cooled in an ice-water bath, 500 ml of 3M hydrochloric acid are added thereto, the temperature being maintained between 5 and 80.degree. C. and then brought to reflux for 45 minutes.

The mixture is cooled again and basified to pH 11 with 30% concentrated sodium hydroxide. The product is extracted with dichloromethane, the organic phase is washed with saturated aqueous sodium chloride solution, dried and the solvent is evaporated off under reduced pressure.

The product is purified by chromatography on silica gel, eluting with dichloromethane.

11.7 g of product, crystallized in hexane, are recovered.

Melting point = 630C.

Analogously - (R) -3-methyl- (3,4-dihydro-2H-1,4-benzoxazin-5-yl) furan-2-yl methanone are obtained; f0] D20 = -28t2 (c = 0.01, methanol).

7-fluoro- (3,4-dihydro-2H-1,4-benzoxazin-5-yl) furan-2-yl methanone;
Melting point = 102 ° C.

8.2. Furan-2-yl (4-nitroso-3,4-dihydro-2H-1,4-benzoxazin-5-yl) methanone.

To a solution of 11.7 g (0.051 mole) of (3,4-dihydro-2H-1,4-benzoxazin-5-yl) furan-2-yl methanone in 400 ml of dichloromethane, cooled with OOC, is rapidly added 4 33 g (0.063 mol) of sodium nitrite dissolved in 60 ml of water. Then a solution of 5.2 g of concentrated sulfuric acid and 60 ml of water is slowly added for 1 hour, keeping the temperature between 0 and 50 ° C. Allowed to warm to room temperature and stirred for 1 hour.

The product is extracted with dichloromethane, the organic phase is washed with water, dried over sodium sulfate and the solvent is evaporated to dryness.

The product is purified by chromatography on silica gel, eluting with dichloromethane.

12.3 g of product are obtained.

Melting point = 1350C.

8.3. 2-Furan-2-yl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine
To a solution of 12.3 g (0.0477 mole) of (3,4-dihydro-2H-1,4-benzoxazin-5-yl) furan-2-yl methanone in 110 ml of glacial acetic acid and 500 ml of methanol, 12.5 g (0.19 mol) of powdered zinc are added in small quantities.

The addition lasts 20 minutes and the temperature of the reaction medium rises to 250C. Allowed to warm to room temperature, the zinc is filtered and the filtrate is concentrated under reduced pressure. The remaining acetic acid is removed by azeotropic evaporation with toluene. The product is extracted with 300 ml of ethyl acetate and the organic phase is washed respectively with 170 ml of 1N hydrochloric acid, 170 ml of 5% sodium hydrogencarbonate, twice with 170 ml of 2N sodium hydroxide and with the water. The organic phase is dried and the solvent is evaporated off under reduced pressure. The product is purified by chromatography on silica gel, eluting with a cyclohexane: ethyl acetate (3: 1) mixture.

8.7 g of product are obtained.

Melting point = 115 C.

Analogously - (R) -8-methyl-2-furan-2-yl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine; [cu] OD -25.8 (c = 0.01, methanol).

4-Fluoro-2-furan-2-yl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine;
Melting point = 122 C.

8.4. 7,8-Dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid.

To a solution of 4 g (0.0177 mol) of 2-furan-2-yl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine in 70 ml of benzene and 70 ml of acetone 8.4 g (0.053 mol) of potassium permanganate are added in small quantities; the temperature of the reaction medium rises to 320C. Stir at room temperature for 3 days. The manganese oxide formed is filtered off and filtered off with dichloromethane. The residue obtained contains 2.26 g of starting material which can be oxidized again. The manganese oxide collected contains the expected acid compound, is taken up with 1.4 M potassium hydroxide, and the suspension is filtered. The filtrate is acidified and the product precipitates. The precipitate is filtered off and 1.11 g of product is recovered. An additional 250 mg of product is recovered by extracting with three times with dichloromethane the filtrate obtained.

Melting point = 2400C.

4-Fluoro-7,8-ddhydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid are obtained in a similar manner.
Melting point = 265 ° C; -Acid (R) -8-methyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid] D20 = -39.2 (c = 0.01, methanol) );
M.p. = 220 ° C;
Enantiomeric purity = 96.84 (HPLC: AGP column).

Example 9: 7,8-Dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid.

9.1. 1- (3,4-Dihydro-2H-1,4-benzoxazin-5-yl) ethanone.

To a solution of 7 g (0.0518 mol) of 3,4-dihydro-2H-1,4-benzoxazine in 75 ml of 1,2-dichloroethane, maintained at a temperature between 0 and 50C and under nitrogen, is added slowly for 1 hour, a solution of 100 ml (0.1 mole) of boron trichloride (1 M in heptane). The mixture is refluxed for 2 hours. It is cooled and added in 20 minutes a solution of 5.36 ml (0.102 mole) of acetonitrile in 15 ml of 1,2-dichloroethane.

The reaction mixture is refluxed for 3 hours and then left overnight at room temperature. In order to hydrolyze the imine intermediate formed, the reaction medium is cooled in an ice-water bath and 500 ml of 3 M hydrochloric acid are added, the temperature being maintained between 5 and 80 ° C. and then brought to reflux for 45 minutes.

The mixture is cooled again and basified to pH 11 with 30% concentrated sodium hydroxide. The product is extracted with dichloromethane, the organic phase is washed with saturated aqueous sodium chloride solution, dried and the solvent is evaporated off under reduced pressure.

The product is purified by chromatography on silica gel, eluting with dichloromethane.

1.6 g of product are recovered in the form of an oil.

Boiling point = 700 ° C (p = 0.6 mm Hg, 4.13 kPa).

9.2. 1- (4-Nitroso-3,4-dihydro-2H-1,4-benzoxazin-5-yl) ethanone.

To a solution, cooled to OOC, 4.75 g (0.0268 mol) of (3,4-dihydro-2H-1,4-benzoxazin-5-yl) ethanone in 70 ml of dichloromethane are rapidly added 2 39 g (0.034 mol) of sodium nitrite dissolved in 30 ml of water. A solution of 2.15 g of concentrated sulfuric acid and 30 ml of water is added slowly over 1 hour while maintaining the temperature between 0 and 5 ° C. The mixture is left to return to ambient temperature and stirred for 1 hour. .

The product is extracted with dichloromethane, the organic phase washed with slurry, dried over sodium sulfate and the solvent evaporated to dryness.

The product is purified by chromatography on silica gel, eluting with dichloromethane.

5.5 g of product are obtained in the form of an oil.

Analogously - (S) 3-Methyl-1- (4-nitroso-3,4-dihydro-2H-1,4-benzoxazin-5-yl) ethanone are obtained; [] D20 = +57.70 (c = 0.01, methanol).

7-fluoro-8-methyl-1- (4-nitroso-3,4-dihydro-2H-1,4-benzoxazin-5-yl) ethanone;
Melting point = 101 ° C.

9.3. 2- (2-Phenylethenyl) -7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine.

1 g (0.0268 mol) of sodium hydroxide is dissolved in 5 ml of water. An ice-cold solution of 5.5 g (0.0268 mol) of 1- (4-nitroso-3,4-dihydro-2H-1,4-benzoxazin-5-yl) ethanone, 2.84 g (0.0268 mol) benzaldehyde, freshly distilled, and 80 ml of absolute ethanol is added dropwise in 20 minutes at 0-50C.

The mixture is stirred at 0-5 ° C for 2 hours, 50 ml of methylene chloride are then added and the mixture is stirred overnight at room temperature. After addition of 200 ml of water, the organic phase collected is dried and the solvent is distilled under reduced pressure.

The product obtained, 1- (4-nitroso-3,4-dihydro-2H-1,4-benzoxazin-5-yl) -3-phenylpro-2-en-1-one, is dissolved in a mixture of 70 ml of methanol and 15 ml of acetic acid. 6 g (0.09 mole) of powdered zinc are carefully added at a temperature between 0 and -50C. The reaction medium is stirred for 1 h 30 min at 0-5 C. 200 ml of methanol are added, filtered on celite and the filtrate is evaporated under reduced pressure. The product is extracted with dichloromethane and washed successively with 1N hydrochloric acid, 2N sodium hydroxide and water. The organic phase is dried and evaporated under reduced pressure. The residue is purified on silica gel eluting with a dichloromethane methanol (99.8: 0.2).

2.7 g of product are obtained in the form of an oil.

Analogously - (S) -8-methyl-2- (2-phenylethenyl) -7,8-dihydropyrazolo [1,5,4de] [1,4] benzoxazine; [Q] D20 = +91.30 (c = 0.01, methanol).

4-Fluoro-8-methyl-2- (2-phenylethienyl) -7,8-ddhydropyrobenz [1,5,4-de] [1,4] benzoxazine [1,4] ] benzoxazine;
Mp = 98 ° C; 4-Fluoro-8-methyl-2- (2-phenylethenyl) -7,8-dihydro-6H-pyrazolo [4,5,1,1] quinoline;
Melting point = 75-76 ° C.

9.4. 7,8-Dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxaldehyde.

This compound is obtained by two methods
Process A
To a solution of 2.7 g (0.0103 mol) of 2- (2-phenylethenyl) 7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine in 100 ml of dichloromethane, cooled to -600C, is bubbled for 1h 15 min, a flow of 2.06 g of ozone. The temperature is slowly allowed to rise to -500 ° C., a stream of nitrogen is bubbled and a solution of 4.32 ml (0.031 mol) of triethylamine in 50 ml of dichloromethane is added. The mixture is stirred for 18 hours, allowing to return to ambient temperature and refluxing the solvent for 3 hours. The product is purified by chromatography on a silica column, eluting with dichloromethane.

0.5 g of product is obtained.

Melting point = 145 ° C.

8-Methyl-7,8-ddhydropyrazolo [1,5,4-de] [1,4] benzoxazine-2carboxaldehyde are obtained analogously;
Melting point = 100 C.

4-Fluoro-8-methyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxaldehyde;
Melting point = 147 ° C.

Method B
5 g (0.019 mol) of 2- (2-phenylethenyl) -7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine are dissolved in 100 ml of ether and 100 ml of water, followed by 1 g (0.0004 mol) of osmium tetroxide are added and the mixture is stirred for 15 minutes.

7.3 g (0.034 mol) of sodium periodate are added in small quantities at room temperature and the mixture is stirred for 2 hours. Again, 0.1 g of osmium tetroxide is added, then 3.6 g (0.0168 mol) of sodium periodate are added in small portions and stirred for 2 hours. The mixture is then washed with water. The ether phase is filtered, dried and evaporated under reduced pressure.

The residue is chromatographed on a silica column, eluting with dichloromethane.

1.75 g of product are obtained.

Analogously - (S) -8-methyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine2-carboxaldehyde; [?] D20 = +94.40 (c = 0.01, methanol);
Melting point = 95 C.

4-Fluoro-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2carboxaldehyde;
Melting point = 1700C.

4-Fluoro-8-methyl-7,8-dihydro-6H-pyrazolo [4,5,1 i] quinoline-2-carboxaldehyde
Melting point = 124 C.

DBP

Melting point = 238 C.

Analogously, 4-fluoro-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid is obtained;
M.p. = 265 ° C; 4-Fluoro-8-methyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid;
M.p. = 242 ° C; 8-methyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid;
Melting point = 195 ° C; - (S) -8-Methyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid; [α] D20 = + 80.20 (c = 0.01, methanol);
Melting point = 187 OC;
Enantiomeric purity = 100% (HPLC: AGP column).

8-phenyl-7,8-dihydropyrazolo [1,5,4-de] [1,4] benzoxazine-2-carboxylic acid;
Mp = 205 ° C 4-Fluoro-8-methyl-7,8-dihydro-6H-pyrazolo [4,5,1-i] quinoline-2-carboxylic acid;
M.p. = 224-225 ° C; 7,8-dihydro-6H-pyrazolo [4,5,1-ij] quinoline-2-carboxylic acid;
Melting point = 210 C.

PREPARATION OF FORMULA (IV) INTERMEDIARIES
Example 9: (S) -3-Methyl-3,4-dihydro-2H-1,4-benzoxazine 9.1. (S) - [2- (2-Bromophenoxy) -1-methylethyl] carbamate 1,1-dimethylethyl.

In a 1 liter three-necked flask containing 300 ml of toluene, 23 g (0.131 mol) of 1,1-dimethylethyl (S) - (2-hydroxy-1-methylethyl) carbamate and 53.9 g (0.200 mol) of triphenylphosphine. The mixture is cooled in an ice bath and 21 ml (0.131 mole) of diethylazodicarboxylate are added dropwise. Stirring is continued for 1 hour, 17.3 ml (0.100 mol) of 2-bromophenol are added dropwise, the mixture is stirred for 2 hours at 0 ° C., the temperature of the reaction medium is allowed to return to ambient temperature and the mixture is stirred overnight. . A precipitate is removed by filtration. The filtrate is then washed with sodium hydroxide 1
N and water, the collected organic phase is dried and concentrated under vacuum. The residue obtained is purified by chromatography on silica gel, eluting with a 70:30 mixture of dichloromethane and heptane.

26.7 g of product are obtained.

 [?] D20 = -40.8 (c = 0.01, methanol).

Is similarly obtained - (R) - [2- (2-Bromophenoxy) -1-methylethyl] carbamate of 1,1 dimethylethyl; [Q] D2O = +39.90 (c = 0.01, methanol).

9.2. 1,1-Dimethylethyl (S) -3-methyl-3,4-dihydro-2H-1,4-benzoxazine-4-methylethylcarbonate.

In a three-necked 1 liter, under an inert atmosphere, 8.38 g (0.087 mole) of sodium tert-butoxide is placed in 150 ml of toluene. 5 g (0.0043 mol) of palladium tetrakis (triphenylphosphine) are added, followed by a solution of 19.28 g (0.06 mol) of (S) - [2- (2-bromophenoxy) -1-methylethyl] carbamate. 1,1-dimethylethyl. The reaction medium is heated to 1100C and allowed to stir for 2h30min.

1 g of palladium tetrakis (triphenylphosphine) is again added and after stirring for 5h30min at 1100C, the reaction medium is allowed to cool and 1 liter of water is added. Stir, allow to settle and extract with toluene. The organic phases are combined, washed with water, dried and the solvent is evaporated off under reduced pressure. It is purified by chromatography on silica gel, eluting with dichloromethane.

10.1 g of product are obtained.

 [?] D20 = +21.20 (c = 0.01, methanol).

Is similarly obtained - (R) -3-methyl-3,4-dihydro-2H-1,4-benzoxazine-4-methylethylcarbonate 1,1-dimethylethyl; [α] 20 = -23.6 (c = 0.01, methanol).

9.3. (S) -3-Methyl-3,4-dihydro-2H-1,4-benzoxazine
Refluxed for 2 hours, 10.1 g (0.04 mol) of dissolved (S) -3-methyl-3,4-dihydro-2H-1,4-benzoxazine-4-methylethyl carbonate of 1,1-dimethylethyl in 60 ml of ethanol and 60 ml of 6N hydrochloric acid. Concentrated under reduced pressure and the residue is basified with sodium bicarbonate. The product is extracted with dichloromethane, the combined organic phases are washed with water, dried and evaporated under reduced pressure.

5.74 g of product are obtained.

 [?] D20 = +67.40 (c = 0.01, methanol).

Is similarly obtained - (R) -3-methyl-3,4-dihydro-2H-1,4-benzoxazine; [α] D20 = -18.8 "(c = 0.01, methanol).

Board

<tb> N <SEP> X <SEP> R1 <SEP> R2 <SEP> R3 <SEP> F <SEP> (C) <SEP>[&alpha;] D20 <SEP><SEP> Salt
<tb> 1 <SEP> CH2 <SEP> 4-H <SEP> H <SEP> H <SEP> 202 <SEP> 1/2 <SEP> fumarate
<tb> 2 <SEP> CH2 <SEP> 4-H <SEP> H <SEP> CH3 <SEP> 226 <SEP> 1/2 <SEP> fumarate
<tb> 3 <SEP> CH2 <SEP> 4-F <SEP> CH3 <SEP> CH3 <SEP> 245 <SEP> oxalate
<tb> 4 <SEP> O <SEP> 4-H <SEP> H <SEP> CH3 <SEP> 154 <SEP> Hydrochloride
<tb> 5 <SEP> O <SEP> 4-F <SEP> H <SEP> CH3 <SEP> 190 <SEP> Hydrochloride
<tb> 6 <SEP> O <SEP> 4-H <SEP> CH3 <SEP> CH3 <SEP> 238 <SEP> oxalate
<tb> 7 <SEP> O <SEP> 4-H <SEP> H <SEP> H <SEP> 226 <SEP> Hydrochloride
<tb> 8 <SEP> O <SEP> 4-F <SEP> CH3 <SEP> H <SEP> 190 <SEP> oxalate
<tb> 9 <SEP> O <SEP> 4-H <SEP> C6H5 <SEP> CH3 <SEP> 140 <SEP> oxalate
<tb> 10 <SEP> O <SEP> 4-F <SEP> CH3 <SEP> CH3 <SEP> 243 <SEP> oxalate
<tb> 11 <SEP> O <SEP> 4-H <SEP> H <SEP> C2H5 <SEP> 132 <SEP> oxalate
<tb> 12 <SEP> O <SEP> 4-H <SEP> (S) -CH3 <SEP> CH3 <SEP> 240 <SEP> +52, <SEP> 5 <SEP> oxalate
<tb> 13 <SEP> O <SEP> 4-H <SEP> (R) <SEP> -CH3 <SEP> CH3 <SEP> 236 <SEP> -30.7 <SEP> 1/2 <SEP> oxalate
<Tb>
The compounds of formula (I) according to the invention have been the subject of pharmacological tests which have shown their interest as active substances in therapeutics.

In particular, they have been tested for their inhibitory effects on the binding of [3H] - (S) -zacopride with serotoninergic receptors of the 5-HT3 type of the rat cortex, according to the method described by NM Barnes et al. J.

Pharm. Pharmacol., 40, 548-551 (1988).

Male Sprague-Dawley rats (OFA, Iffa credo) of 200 to 250 g are euthanized and their brain is removed. The cortex is dissected and homogenized using a PolytronX mill (position 7.20 s) in 20 volumes of 25 mM Tris buffer (pH = 7.4, 220C). The homogenate is centrifuged for 10 min at 45000xg (in a SORVALL centrifuge equipped with an SS34 rotor), then the pellet is resuspended in 10 volumes of Tris buffer and incubated at 37 ° C. for 10 min with stirring. then the suspension at 20 volumes using Tris buffer and centrifuged under the same conditions as above The pellet obtained is resuspended in 5 volumes of Tris buffer and then divided into 5 ml aliquots which are frozen at -800C On the day of the experiment, the preparation is thawed at 40 ° C. and then diluted 1.2 times using the Tris-NaCl incubation buffer (25 mM Tris, 150 mM NaCl, pH = 7.4, 220 ° C.).

The membrane suspension (100 μl, 1 mg protein) is incubated at 25 ° C for 25 min in the presence of 0.5 nM [3 H] (S) -zacopride (specific activity: 75-85 Ci / mmole, Amersham,
Little Chalfont, UK) in a final volume of 500 μl of Tris-NaCl buffer, in the absence or in the presence of the test compound.

Incubation is stopped by filtration using filters
Whatman GF / B pretreated with polyethylenimine (0.1 a). Each reaction tube is prediluted with 4 ml of Tris-NaCl buffer and then rinsed 3 times with 4.5 ml of Tris buffer.
NaCl.

The filters are pre-cut before drying in the oven (1200C, 5 min). The radioactivity retained on the filters is measured by liquid scintigraphy. Nonspecific binding is determined in the presence of 10 μM of MDL 72222.

For each concentration of test compound, the percent inhibition of the specific binding of [3H] - (S) zacopride is determined, followed by the concentration of the inhibiting compound 50% of the specific binding of [3H] - (S) -zacopride. (IC50).

The IC50's of the compounds of the invention are between 1 nM and 2 μM.

The compounds of the invention have also been studied for their affinity for 5-HT 4 receptors in the guinea pig striatum, according to the method described by Grossman et al.
Br. J. Pharmacol., 109, 618-624 (1993).

Guinea pigs (Hartley, Charles River) were euthanized from 300 to 400 g and their brain removed. The striata are excised and frozen at -80 ° C. On the day of the experiment, the tissue is thawed at + 40 ° C. in 33 volumes of 50 mM Hepes-NaOH buffer (pH = 7.4 at 20 ° C.) homogenized using a PolytronB mill The homogenate is centrifuged for 10 min at 48,000 × g, the pellet is recovered, resuspended and centrifuged again under the same conditions. Hepes-NaOH buffer (30 mg fresh tissue / ml) This membrane suspension is used as it is.

100 μl of the membrane suspension are incubated with OOC for 120 minutes, in the presence of 0.1 nM of [3 H] GR113808 (specific activity: 80-85 Ci / mmol), in a final volume of 1 ml of Hepes-NaOH buffer. (50 mM, pH = 7.4), in the absence or in the presence of the test compound. The incubation was stopped by filtration on Whatman GF / BS filters, previously treated with 0.1% polyethyleneimine, each tube was rinsed with 4 ml of buffer at 0 ° C. and filtered again. The radioactivity retained on the filters is measured by liquid scintigraphy. Nonspecific binding was determined in the presence of serotonin MA.

The specific binding represents 90E of the total radioactivity recovered on the filter.

For each concentration of compound studied, the percent inhibition of the specific binding of [3 H] GR113808 and the concentration of the test compound which inhibits 50% of the specific binding (IC 50) are determined.

The IC50's of the compounds of the invention are between 100 nM and 2 WM.

Finally, the compounds of the invention have been studied as to their agonist or antagonistic effects on 5-HT4 receptors in the rat esophagus according to the method described by
Baxter et al. In Naunyn Schmied Arch. Pharmacol. (1991), 343, 439.

Male Sprague Dawley rats weighing from 300 to 450 g are used.

A fragment of about 1.5 cm from the end part of the esophagus is rapidly removed, the muscular layer is removed, the inner muscular mucous membrane is opened longitudinally and mounted in an isolated organ vessel containing a Krebs solution. -Henseleit at 320C oxygenated with a carbogenous current (95% O2 and 5% CO2), and connected to an isometric transducer at a basal tension of 0.5 g.

The compounds are studied in increasing cumulative concentrations, from 0.1 nmol / l to 10 pmol / l, and their relaxing effects are compared with that of serotonin (1 μmol / l), chosen as reference agonist.

The compounds of the invention are active in this test.

The results of the biological tests show that the compounds of formula (I) according to the invention are 5-HT3 and 5-HT4 serotoninergic receptor antagonists.

They can therefore be used for the preparation of drugs intended to act as antagonists of the 5-HT3 and 5-HT4 serotoninergic receptors.

The compounds of the invention of formula (I), are particularly useful for the treatment and / or prevention of intestinal functional disorders such as diarrhea, esophageal reflux, intestinal motor disorders, disorders of intestinal secretion, colon irritable, viscerosensitivity; cystic fibrosis of the pancreas, carcinoid syndrome; urinary or intestinal incontinence, megacolon.

The compounds of the invention are still useful for the treatment and / or prevention of nausea and vomiting, for example following antitumor treatment or administration of anesthetic; oesophageal spasm; central nervous system disorders such as schizophrenia, mania, anxiety and depression; cognitive disorders such as senile or presenile Alzheimer's dementia; dyskinesia, pain, migraines and headaches; addiction or withdrawal disorders of alcohol or drugs; disorders of gastrointestinal function such as dyspepsia, peptic ulcer, heartburn, flatulence; disorders of the cardiovascular system and respiratory disorders.

The compounds of the invention, in combination with suitable excipients, may be presented in any form suitable for oral or parenteral administration, such as tablets, dragees, capsules, capsules, suspensions or oral or injectable solutions, and dosed to allow administration of 0.005 to 5 mg / kg, 1 to 4 times daily.

Claims (4)

claims 1. Compounds of general formula (I)

 in which X represents an oxygen atom or a methylene group, R1 represents a hydrogen atom, a halogen such as chlorine or fluorine, a C14 alkoxyl group, or a hydroxyl or amino group, and R2 and R3, which may be identical or different, represent either a hydrogen atom or a linear or branched C14 alkyl group, a phenyl group or a benzyl group, in the form of enantiomers, of diastereoisomers, including racemic mixtures, and their addition salts with pharmaceutically acceptable acids. 2. Process for the preparation of the compounds according to claim 1, characterized in that a compound of formula (II) is reacted

 wherein R1 and R2 are as defined in claim 1, with a compound of formula (III)

 wherein R3 is as defined in claim 1. 3. Medicinal product characterized in that it consists of a compound according to claim 1. 4. A pharmaceutical composition characterized in that it comprises a compound according to claim 1 and one or more excipients.