FR2750427A1 - 1-OXO-1, 2-DIHYDROISOQUINOLINE-4-PROPANAMIDE DERIVATIVES, THEIR PREPARATION AND THEIR THERAPEUTIC USE - Google Patents

Abstract

The invention discloses compounds of general formula (I) in which X represents an atom of hydrogen or of chlorine or of a methyl group, trifluoromethyl or methoxy; R1 represents a methyl group; R2 and R3 each represent an atom of hydrogen or an alkyl group, or form, with the nitrogen atom, a pyrrolidinyl ring; and R4 represents a furan-2-yl, thien-2-yl, thien-3 yl or 5-methyl-thien-3-yl. It is applicable in therapeutics.

Description

The present patent application relates to 1-oxo-1,2-dihydroisoquinoline-4-propanamide derivatives, their preparation and their therapeutic application.

dans laquelle
X représente un atome d'hydrogène ou de chlore ou un groupe méthyle ou trifluorométhyle,
R1 représente un groupe méthyle,
R2 représente un groupe méthyle,
R3 représente un atome d'hydrogène, et
R4 représente soit un groupe furan-2-yle, soit un groupe thién-2-yle ou thién-3-yle éventuellement substitué en position 5 par un groupe méthyle.The compounds of the invention correspond to the general formula (I)

in which
X represents a hydrogen or chlorine atom or a methyl or trifluoromethyl group,
R1 represents a methyl group,
R2 represents a methyl group,
R3 represents a hydrogen atom, and
R4 represents either a furan-2-yl group, a thien-2-yl or thien-3-yl group optionally substituted in the 5-position by a methyl group.

According to the invention, the compounds of general formula (I) can be prepared according to a process illustrated by the following scheme.

dans un solvant éthéré tel que le tétrahydrofurane, à une température de 20 à 650C, ou bien avec le (triphénylphosphoranylidène)acétate de méthyle, dans un solvant aromatique tel que le toluène, à une température de 20 à ll00C, pour obtenir un ester de formule générale (VII), que l'on soumet à hydrogénation, par voie catalytique ou chimique, par exemple avec le borohydrure de sodium en présence de chlorure de cuivre, pour obtenir un ester de formule générale (VIII), et finalement on traite ce dernier avec une amine de formule générale HNR2R3, à température ambiante et dans un solvant tel qu'un mélange de méthanol et de dichlorométhane.An amide of the general formula (II), in which R 1 and R q are as defined above, is reacted with thionyl chloride to give a chlorimine of the general formula (III), which is reacted with a anhydride of general formula (IV), wherein X is as defined above, in an aromatic solvent, for example toluene, at a temperature of 70 to 1100C, to obtain an isoquinolinone of general formula (V), which is treated with phosphorus oxychloride, in the presence of N, N-dimethylformamide, at a temperature of 20 to 80 ° C, to obtain, after hydrolysis, an aldehyde of general formula (VI), and then the latter is treated with dimethoxyphosphinyl) methyl acetate, Scheme

in an ethereal solvent such as tetrahydrofuran, at a temperature of 20 to 650C, or with methyl (triphenylphosphoranylidene) acetate, in an aromatic solvent such as toluene, at a temperature of 20 to 100C, to obtain an ester of general formula (VII), which is subjected to hydrogenation, catalytically or chemically, for example with sodium borohydride in the presence of copper chloride, to obtain an ester of general formula (VIII), and finally this is treated last with an amine of general formula HNR2R3, at room temperature and in a solvent such as a mixture of methanol and dichloromethane.

Amides of general formula (II) can be prepared by methods such as those described in Tetrah. Lett.

(1994) 4149-4166 and J. Am. Chem. Soc. (1962) 84 3410.

The anhydride of the general formula (IV) is commercially available when X is hydrogen and the others can be prepared by methods such as those described in Arch. Pharm. (1991) 324-508-518.

The examples which follow illustrate the preparation of some compounds of the invention. Elemental microanalyses, and I.R. and R.M.N. confirm the structures of the compounds obtained.

The numbers indicated in parentheses in the titles of the examples correspond to those in the 1st column of Table 1 given below.

In the names of the compounds, the dash - is part of the word, and the dash "" is only used for the cut at the end of the line it is to be deleted in the absence of cut, and must not be replaced by a normal dash nor by a space.

Example 1 (Compound No. 2)

 V-Chloro-N, 2-dimethyl-1-oxo-3-thien-2-yl-1,2-dihydroisoquinoline-4-propanamide.

1.1. 7-Chloro-2-methyl-3-thien-2-ylisocuinoléin-1 (2H) -one.

Under a nitrogen atmosphere, 40.77 g (342 mmol) of thionyl chloride are added to 5.20 g (36.8 mmol) of N-methyl-thiophene-3-carboxamide and the mixture is heated at reflux for 2:30.

The medium is concentrated under reduced pressure, 100 ml of toluene and then 7.24 g (36.8 mmol) of 7-chlorophthalic anhydride are rapidly added to the residue, the mixture is gradually heated to reflux and heating is continued for 3 hrs.

The mixture is allowed to return to room temperature, it is concentrated under reduced pressure and the residue is purified by chromatography on a column of silica gel, eluting with cyclohexane and then with a mixture of cyclohexane and dichloromethane ranging from 90/10 to 50 / 50, then with dichloromethane.

10.13 g (36.7 mmol) of beige solid are obtained.

Melting point: 115-1160C.

1.2. Chloro-2-methyl-l-oxo-3-thien-2-yl-2-dihydroisoqui
quinoline-4-carboxaldehyde.

Under a nitrogen atmosphere, 50 g (0.68 mol) of
Dry N, N-dimethylformamide at 0 ° C. 20.24 g (191 mmol) of phosphorus oxychloride are then added dropwise, then at room temperature 10.13 g (36.7 mmol) of toluene are added. chloro-2-methyl-3-thien-2-ylisoquinolin-1 (2H) -one and the mixture is heated at 80 ° C for 2 h.

The mixture is allowed to cool, poured onto 200 g of crushed ice, 30% aqueous sodium hydroxide is added until pH = 10, the mixture is cooled, and the mixture is diluted to 400 ml with water. the extract with 2 times 150 ml of dichloromethane, the organic phase is washed with 4 times 250 ml of water, dried over sodium sulfate, filtered and the solvent evaporated under reduced pressure. The residue is purified by chromatography on a column of silica gel, eluting with cyclohexane alone, then with an 80/20 mixture of cyclohexane and dichloromethane, then with a 50/50 mixture of cyclohexane and dichloromethane, followed by dichloromethane alone. After evaporation, 8.19 g (27 mmol) of yellowish solid are obtained.

Melting point: 166-167 ° C.

1.3. (E) -3- (7-Chloro-2-methyl-1-oxo-3-thien-2-yl-l, 2-
methyl dihydroisoquinolin-4-yl) prop-2-enoate.

In a three-necked flask of 500 ml placed under an argon atmosphere, 1.6 g (40 mmol) of sodium hydride in suspension at 60% in the oil are introduced, washed with pentane, suspended. in 200 ml of dry tetrahydrofuran, cooled to 0 ° C., 6.0 g (33 mmol) of methyl (dimethoxyphosphinyl) acetate dissolved in 20 ml of tetrahydrofuran and then 8.19 g (27 ml) are added dropwise. mmol) of 7-chloro-2-methyl-1-oxo-3-thien-2-yl-1,2-dihydroisoquinoline-4-carboxaldehyde and the mixture is refluxed for 3 h.

It is allowed to cool, 15 ml of methanol are slowly added to destroy the excess hydride, the solvent is evaporated off under reduced pressure, the residue is taken up in 200 ml of dichloromethane, 100 ml of hydrochloric acid 0 are added, 1M, the organic phase is separated off, washed with water, dried over sodium sulfate, filtered, the solvent is evaporated off under reduced pressure and the residue is purified by chromatography on a silica gel column. eluting with a 50/50 mixture of cyclohexane and dichloromethane, then with dichloromethane, then with a 99/1 mixture of dichloromethane and ethyl acetate, then with a 95/5 mixture of dichloromethane and acetate of ethyl. 8.71 g of orange-colored solid is obtained.

Melting point: 161-162 ° C.

1.4. 7-Chloro-2-methyl-1-oxo-3-thien-2-yl-1,2-dihydroisoquine
methyl nolein-4-propanoate.

In a 500 ml three-necked flask containing 8.71 g (24.2 mmol) of (E) -3- (7-chloro-2-methyl-1-oxo-3-thien-2-yl) -1,2- Methyl dihydroisoquinolin-4-yl) prop-2-enoate is admixed with 7.2 g (72.7 mmol) of cuprous chloride and 220 ml of methanol. Under a nitrogen atmosphere, the mixture is cooled with a water-ice bath and 8.5 g (225 mmol) of sodium borohydride are introduced in small portions and the mixture is stirred at room temperature for 2 hours. .

The mixture is filtered on diatomaceous earth, the solvent is evaporated off under reduced pressure, 1 M hydrochloric acid is added to the residue until pH = 1, the mixture is extracted with 250 ml of dichloromethane, the organic phase is washed with the water, dried over sodium sulfate, filtered and the solvent evaporated under reduced pressure. The residue is purified by chromatography on a column of silica gel, eluting with dichloromethane and then a mixture of dichloromethane and ethyl acetate ranging progressively from 99/1 to 95/5.

7.8 g of a mixture of the desired product and of the starting product are obtained, reduced again in 150 ml of methanol with 3.0 g (30.3 mmol) of cuprous chloride and 4.5 g (30.3 mmol) of cuprous chloride. g (119 mmol) of sodium borohydride.

After treatment, the crude product is purified by chromatography on a column of silica gel, eluting with a 1/1 mixture of cyclohexane and dichloromethane, then with dichloromethane and then with a mixture ranging from 99/1 to 90/10 of dichloromethane and of ethyl acetate. After recrystallization from cyclohexane, 5.05 g (14 mmol) of white solid are obtained.

Melting point: 121.5-122.5 ° C.

1.5. 7-Chloro-N, 2-dimethyl-l-oxo-3-thien-2-yl-l, 2-dihydro
isoquinoline-4-propanamide.

30 ml of dichloromethane and then 230 ml of methanol are added to 1.4 g (3.9 mmol) of 7-chloro-2-methyl-1-oxo-3-thien-2-yl 1,2-dihydroisoquinoline-4. methyl propanoate, and gaseous methylamine is introduced into the resulting solution until saturation.

After 2 days, the volatile compounds are evaporated off under reduced pressure, 200 ml of dichloromethane and 100 ml of 0.1 M hydrochloric acid are added to the residue, the organic phase is separated, washed with water and dried over sodium sulfate, filtered and the solvent evaporated under reduced pressure.

The residue is purified by chromatography on a column of silica gel, eluting with a mixture of dichloromethane and ethyl acetate ranging progressively from 90/10 to 50/50, then with a mixture of dichloromethane and methanol ranging progressively from 95.degree. / 5 to 90/10.

Finally, 0.97 g (2.7 mmol) of solid is obtained.

Melting point: 191.0-192.5 ° C.

Example 2 (Compound No. 6)

7-Chloro-N, 2-dimethyl-1-oxo-3-thien-3-yl-1,2-dihydroisoquinoline-4-propanamide.

2.1. 7-Chloro-2-methyl-3-thien-3-ylisoquinoléin-1 (2H) -one.

16.3 g (137 mmol) of thionyl chloride are added to 3.60 g (25.9 mmol) of N-methylthiophene-3-carboxamide and then, under a nitrogen atmosphere, the mixture is refluxed for 1 hour. .

The mixture is allowed to cool, 100 ml of toluene and then 5 × 10 g (26 mmol) of 7-chlorophthalic anhydride are rapidly added to the residue, the mixture is gradually heated to reflux and the heating is continued for 4 hours.

The mixture is allowed to return to room temperature, it is concentrated under reduced pressure and the residue is purified by chromatography on a column of silica gel, eluting with a mixture of cyclohexane and dichloromethane ranging progressively from 90/10 to 50/50, and then with dichloromethane alone and then with a mixture of dichloromethane and ethyl acetate ranging progressively from 99/1 to 95/5.

After recrystallization from cyclohexane, 4.67 g (16.9 mmol) of yellow crystalline solid are obtained.

Melting point: 115-116 ° C.

2.2. 7-Chloro-2-methyl-1-oxo-3-thien-3-yl-1,2-dihydroisoqui
quinoline-4-carboxaldehyde.

Under a nitrogen atmosphere 40 ml of dry N, N-dimethylformamide are cooled to 0.degree. C., 11.5 g (75.1 mmol) of phosphorus oxychloride are then added dropwise and then, at room temperature, 3.0 g (11.6 mmol) of 7-chloro-2-methyl-3-thien-3-ylisoquinolin-1 (2H) -one are added and the mixture is heated at 60 ° C for 5 h.

The mixture was allowed to cool, poured onto 150 g of crushed ice, 30% aqueous sodium hydroxide was added to pH = 10, the mixture was cooled, and the mixture was diluted to 300 ml with slug. extracted with 350 ml of dichloromethane, the organic phase is washed with water, dried over sodium sulfate, filtered and the solvent evaporated under reduced pressure. The residue is purified by chromatography on a column of silica gel, eluting with cyclohexane alone and then with a mixture ranging progressively from 80/20 to 50/50 of cyclohexane and dichloromethane, then with a mixture of dichloromethane and dichloromethane. ethyl progressively from 99/1 to 98/2. 1.01 g (3.3 mmoles) of an oil which slowly solidifies is obtained.

2.3. 3- (7-Chloro-2-methyl-1-oxo-3-thien-3-yl-1,2-dihydro ~
methyl isoquinolin-4-yl) prop-2-enoate.

Under a nitrogen atmosphere, a mixture of 1.01 g (3.3 mmol) of 7-chloro-2-methyl-1-oxo-3-thien-3-yl-1,2-dihydroisoquinoline-4 is refluxed. -carboxaldehyde 80 ml of toluene and 1.50 g (4.5 mmol) of methyl (triphenylphosphoranylidene) acetate for 8 h.

It is allowed to cool to ambient temperature, the solvent is evaporated off under reduced pressure, and the residue is purified by chromatography on a column of silica gel, eluting with cyclohexane alone and then with a mixture ranging progressively from 80/20 to 50/50 of cyclohexane and dichloromethane, then with dichloromethane alone, then with a mixture of dichloromethane and ethyl acetate ranging progressively from 99/1 to 95/5. 1.31 g of compound are obtained in the form of a mixture (E) and (Z), which is used as such in the next step.

2.4. 7-Chloro-2-methyl-1-oxo-3-thien-3-yl-1,2-dihydroisoqui ~
methyl nolein-4-propanoate.

In a 3-necked 250 ml flask, 1.31 g (3.3 mmol) of 3- (7-chloro-2-methyl-1-oxo-3-thien-3-yl-1,2-dihydroisoquinolin-4-yl) are introduced. ) methyl prop-2-enoate, 1.0 g (10.1 mmol) of cuprous chloride, 50 ml of tetrahydrofuran and 50 ml of methanol, the mixture is placed under an argon atmosphere and in a water bath and of ice, 1.4 g (37 mmol) of sodium borohydride are added in small portions, the cold bath is removed and the mixture is stirred at room temperature for 2 hours.

It is filtered through diatomaceous earth, the filtrate is concentrated under reduced pressure, the residue is acidified with 1M hydrochloric acid to pH = 1 and extracted with 250 ml of dichloromethane. The organic phase is washed with water, dried over sodium sulfate, filtered, the solvent is evaporated off under reduced pressure, and the residue is purified by chromatography on a column of silica gel, eluting with a mixture ranging from 10/90 to 50/50 of cyclohexane and dichloromethane, then with dichloromethane, then with a mixture of dichloromethane and ethyl acetate ranging from 99/1 to 90/10. 0.67 g (1.86 mmol) of pasty product is obtained which is used as such in the next step.

2. 5. 7-Chloro-N, 2-dimethyl-1-oxo-3-thien-3-yl-1,2-dihydro
isoquinoline-4-propanamide.

0.67 g (1.86 mmol) of methyl 7-chloro-2-methyl-1-oxo-3-thien-3-yl-1,2-dihydroisoquinoline-4-propanoate are dissolved in a mixture of 20 ml. of dichloromethane and 100 ml of methanol, the solution obtained is saturated with gaseous methylamine and the mixture is stirred for 2 days.

The mixture is concentrated under reduced pressure, 150 ml of water and 150 ml of dichloromethane are added to the residue, the organic phase is separated off, washed with water, dried over sodium sulphate, filtered and filtered. the solvent is evaporated off under reduced pressure and the residue is purified by chromatography on a column of silica gel, eluting with dichloro-methane alone and then with a mixture of dichloromethane and methanol ranging progressively from 98/2 to 80/20. 0.40 g of white amorphous solid is obtained.

Melting point: 167.5-168.5 ° C.

Example 3 (Compound No. 7)

7-Chloro-3-furan-2-yl-N, 2-dimethyl-1-oxo-1,2-dihydroisoquinoline-4-propanamide.

3.1. 7-Chloro-3-furan-2-yl-2-methylisoquinolin-1 (2H) -one.

Under a nitrogen atmosphere, 40.8 g (343 mmol) of thionyl chloride are added to 10.0 g (79.9 mmol) of N-methyl-furan-2-carboxamide, and the mixture is refluxed 2:30.

The medium is concentrated under reduced pressure, 50 ml of toluene and then 10.4 g (52.9 mmol) of 7-chlorophthalic anhydride are rapidly added to the residue, the mixture is gradually heated to reflux and heating is continued for 3 hrs.

The mixture is allowed to return to room temperature, it is concentrated under reduced pressure and the residue is purified by chromatography on a column of silica gel, eluting with dichloromethane alone and then with a mixture of dichloro-methane and ethyl acetate. going progressively from 98/2 to 70/30.

After crystallization from cyclohexane, 7.83 g (28.4 mmol) of beige solid are obtained.

Melting point: 127.0-128.5 ° C.

3.2. 7-Chloro-3-furan-2-yl-2-methyl-1-oxo-1,2-dihydroisOqui ~
quinoline-4-carboxaldehyde.

Under a nitrogen atmosphere 30 ml of dry N, N-dimethylformamide are cooled to 0 ° C., 19.75 g (129 mmol) of phosphorus oxychloride are added dropwise, the cold bath is removed, 7.10 g (27.3 mmol) of 7-chloro-3-furan-2-yl-2-methylisoquinolin-1 (2H) -one are added and the mixture is heated at 80 ° C for 2 h 30 min.

The mixture was allowed to cool, poured onto 150 g of crushed ice, 30% aqueous sodium hydroxide was added to pH = 10, the mixture was cooled, and the mixture was diluted to 400 ml with water. the extract with 2 times 150 ml of dichloromethane, the organic phase is washed with water, dried over sodium sulfate, filtered and the solvent evaporated under reduced pressure. The residue is purified by chromatography on a column of silica gel, eluting with cyclohexane alone, then with a mixture ranging progressively from 80/20 to 50/50 of cyclohexane and dichloromethane, then with dichloromethane alone and then with a mixture of progressively from 98/2 to 80/20 of dichloromethane and ethyl acetate. 3.34 g (11.9 mmol) of yellow solid are obtained.

Melting point: 188-189.5 ° C.

3.3. (E) -3- (7-Chloro-3-furan-2-yl-2-methyl-1-oxo-1,2-
methyl dihydroisoquinolin-4-yl) prop-2-enoate.

In a 500 ml three-necked flask placed under an argon atmosphere, 0.8 g (20 mmol) of sodium hydride in a 60% suspension in the oil are introduced, washed with pentane and suspended. in 200 ml of dry tetrahydrofuran, cooled to OOC, 3.28 g (18 mmol) of methyl (dimethoxyphosphinyl) acetate dissolved in 15 ml of tetrahydrofuran and then 3.34 g (11.9 g) are added dropwise. mmol) of 7-chloro-3-furan-2-yl-2-methyl-1-oxo-1,2-dihydroisoquinoline-4-carboxaldehyde and the mixture was refluxed for 6 h.

It is allowed to cool, 5 ml of methanol are slowly added to destroy the excess of sodium hydride, the solvent is evaporated off under reduced pressure, the residue is taken up in 250 ml of dichloromethane, 150 ml of hydrochloric acid are added. 1 M, the organic phase is separated, washed with water, dried over sodium sulfate, filtered, the solvent is evaporated under reduced pressure, the residue is purified by chromatography on a column of silica gel eluting with dichloromethane alone and then with a mixture ranging progressively from 99/1 to 80/20 of dichloromethane and ethyl acetate. 2.39 g (7.2 mmol) of solid are obtained.

Melting point: 121-122 ° C.

3.4. 7-Chloro-3-furan-2-yl-2-methyl-1-oxo-1,2-dihydroisoqui
methyl nolein-4-propanoate.

In a 500 ml three-necked flask containing 2.49 g (7.24 mmol) of (E) -3- (7-chloro-3-furan-2-yl-2-methyl-1-oxo-1,2-dihydroisoquinoline) 4-yl) methyl prop-2-enoate is added 1.3 g (13.1 mmol) of cuprous chloride and 50 ml of tetrahydrofuran and 100 ml of methanol. Under a nitrogen atmosphere, the mixture is cooled with a water-ice bath, 6.5 g (172 mmol) of sodium borohydride are introduced in small portions and the mixture is stirred at room temperature for 2 hours. .

The mixture is filtered on diatomaceous earth, the solvent is evaporated off under reduced pressure, the residue is added ice and 1 M hydrochloric acid to pH = 5, the mixture is extracted with 250 ml of dichloromethane, and the mixture is washed. organic phase with water, dried over sodium sulfate, filtered and the solvent evaporated under reduced pressure. The residue is purified by chromatography on a column of silica gel, eluting with dichloromethane alone and then with a mixture ranging progressively from 99/1 to 50/50 of dichloromethane and ethyl acetate.

0.44 g (1.27 mmol) of solid are obtained.

Melting point: 65-68 ° C.

3.5. 7-Chloro-3-furan-2-yl-N, 2-dimethyl-1-oxo-l, 2-dihydro
isoquinoline-4-propanamide.

Methyl 7-chloro-3-furan-2-yl-2-methyl-1-oxo-1,2-dihydroisoquinoline-4-propanoyl (0.44 g, 1.27 mmol) was dissolved in 15 ml of dichloromethane and 120 ml of methyl chloride. ml of methanol and saturated the resulting solution with methylamine gas at room temperature for 12 h.

The mixture is concentrated under reduced pressure, 150 ml of dichloromethane and 100 ml of 1M hydrochloric acid are added to the residue, the organic phase is washed with water, dried over sodium sulphate and filtered. the solvent is evaporated off under reduced pressure, the residue is purified by chromatography on a column of silica gel, eluting with a mixture of dichloromethane and ethyl acetate ranging progressively from 90/10 to 50/50, and then with a mixture of dichloromethane and methanol ranging progressively from 95/5 to 90/10. After recrystallization from a small volume of ethyl acetate, 0.4 g of white amorphous solid is obtained.

Melting point: 199.0-199.5 ° C.

The following table illustrates the chemical structures and the physical properties of some compounds of the invention.

In the R4 "2-C4H3S" column denotes a thien-2-yl group, "2-C4H3S-5-CH3" denotes a 5-methylthien-2-yl group, "3-C4H3S" denotes a thien-3-thienyl group. yl, and "2-C4H30" refers to a furan-2-yl group.

Board

<tb> N <SEP> X <SEP> R1 <SEP> R2 <SEP> R3 <SEP> R4 <SEP> | <SEP> F <SEP> (C) <SEP>
<tb><SEP> 1 <SEP> H <SEP> CH3 <SEP> CH3 <SEP> H <SEP> 2-C4H3S <SEP> 206.5-207.5
<tb><SEP> 2 <SEP> 7-Cl <SEP> CH3 <SEP> CH3 <SEP> H <SEP> 2-C4H3S <SEP> 191-192.5
<tb><SEP> 3 <SEP> 7-CH3 <SEP> CH3 <SEP> CH3 <SEP> H <SEP> 2-C4H3S <SEP> 186.5-187.5
<tb><SEP> 4 <SEP> 7-CF3 <SEP> CH3 <SEP> CH3 <SEP> H <SEP> 2-C4H3S <SEP> 183-184
<sep> 5 <SEP> 7-Cl <SEP> CH3 <SEP> CH3 <SEP> H <SEP> 2-C4H3S-5-CH3 <SEP> 183-184
<tb><SEP> 6 <SEP> 7-Cl <SEP> CH3 <SEP> CH3 <SEP> H <SEP> 3-C4H3S <SEP> 167.5-168.5
<tb><SEP> 7 <SEP> 7-Cl <SEP> CH3 <SEP> CH3 <SEP> H <SEP> 2-C4H3O <SEP> 199-199.5
<Tb>
The compounds of the invention have been subjected to pharmacological tests which have demonstrated their interest as substances with therapeutic activities.

Study of the membrane binding with respect to a # (benzodiazepine) receptor population associated with GA receptors containing subunit 5.

These receptors may be selectively labeled in rat hippocampus membranes incubated in the presence of [3H] flumazenil and 5M zolpidem (to mask other o receptor subtypes).

The compounds have been studied in vitro for their affinity for these [3H] flumazenil-labeled receptors.

The animals used are male OFA rats (Iffa Credo) of 200 to 250 g. After decapitation, the hippocampus is removed and crushed using an Ultra-Turraxm or PolytronE apparatus for 20 seconds at 6/10 of the maximum speed in 80 volumes of 50 mM Tris buffer at a pH adjusted to 7.4 with hydrochloric acid, and containing 120 mM sodium chloride and 5 mM potassium chloride (5 mM).

The binding with [3H] flumazenil (1 nM, specific activity: 80-87 Ci / mmol, Du Pont de Nemours / New England
Nuclear) is determined by incubating 200 μl of membrane suspension in a final volume of 1 ml buffer containing 5 am of zolpidem and the test compound. After incubation for 45 minutes at 00 ° C., the membranes are recovered by filtration on Whatman GF / BE filters which are washed twice with 5 ml of ice-cold buffer. The amount of radioactivity retained by the filter by liquid scintigraphy is measured.

The specific binding of [3H] flumazenil is defined as the amount of radioactivity retained on the filters and can be inhibited by co-incubation with 1 AM flunitrazepam.

For each concentration of test compound, the percent inhibition of [3 H] flumazenil binding is determined, followed by the IC 50 concentration, which inhibits 50% of the specific binding.

The most active compounds of the invention in this test have an IC 50 of the order of 1 to 30 nM.

Study of the membrane bonds with respect to the 2 (benzodiazepine type II) receptors associated with the GABAA receptors mainly containing the alpha 2 and α 3 subunits.

The affinity of the compounds for the spinal cord receptors 22 was determined according to a variant of the method described by S. Z. Langer and S. Arbilla in Fund. Clin.

Pharmacol. (1988) 2 159-170, using [3H] flumazenil instead of [3H] diazepam as radioligand.

The tissue of the spinal cord is homogenized for 60 seconds in 30 volumes of ice-cold buffer (50 ml Tris / HCl, pH 7.4, 120 mM NaCl, 5 mM KCl) and, after dilution to 1/3, incubation is carried out. suspension with [3H] flumazenil (specific activity 78 Ci / mmol, New England Nuclear) at a concentration of 1 nM and with the compounds of the invention, at different concentrations, in a final volume of 525 μl. After incubating for 30 min at 0 ° C., the samples were filtered under vacuum on Whatman GF / BTM filters and washed immediately with ice-cold buffer. The specific binding of [3H] flumazenil is determined in the presence of unlabeled DM diazepam. The data are analyzed according to the usual methods and the IC50, a concentration which inhibits 50E of [3H] flumazenil binding, is calculated.

The IC.sub.50 of the compounds of the invention are in this test between 30 and 150 nM.

Study of membrane bonds with respect to receptors 1 (benzodiazepine type I) associated with GABAA receptors containing the a1 subunit.

The affinity of the compounds for the receptors 1 of the cerebellum was determined according to a variant of the method described by
SZ Langer and S. Arbilla in Fund. Clin. Pharmacol. (1988) 2 159-170, using [3H] flumazenil instead of [3H] diazepam as radioligand.

Cerebellar tissue was homogenized for 60 seconds in 120 volumes of ice-cold buffer (50 ml Tris / HCl, pH 7.4, 120 mM NaCl,
5 mM KCl) and, after dilution to 1/3, the suspension is incubated with [3 H] flumazenil (specific activity: 78
Ci / mmole; New England Nuclear) at a concentration of 1 nM and with the compounds of the invention, at different concentrations, in a final volume of 525 μl. After incubating for 30 min at 0 ° C, the samples were filtered under vacuum on Whatman GF / BE filters and washed immediately with ice-cold buffer. The specific binding of [3H] flumazenil is determined in the presence of unlabeled 1 M diazepam. The data are analyzed according to the usual methods and the IC50, a concentration which inhibits 50 of [3 H] flumazenil binding, is calculated.

The IC.sub.50 of the compounds of the invention are in this test between 70 and 200 nM.

The results of the tests carried out on the compounds of the invention show that, in vitro, they selectively displace [3H] flumazenil from its membrane binding sites with respect to a population of o (benzodiazepine) receptors associated with GABAA receptors. containing the a5 subunit, compared to the subunit # 1 subtypes associated with GABAA receptors containing the subunit

The compounds of the invention can be used in the treatment of disorders related to disorders of GABAergic transmission in general, such as anxiety, sleep disorders, epilepsy, and particularly related to GABAA receptors associated with the subtype. -unit a5. The preferential distribution of oe receptors, associated with the aS subunit of the GABAA receptor complex, in the olfactory bulb, in limbic structures such as the hippocampus and hypothalamus, and in the spinal cord, suggests that the compounds of the The invention can be used in the treatment of olfactory disorders, cognitive disorders, hormonal disorders related to hypothalamic dysfunction, certain emotional disorders and pain perception.

They can also be used in the treatment of spasticity and muscle contractures.

For this purpose they may be presented in all galenic forms, combined with appropriate excipients, for enteral or parenteral administration, for example in the form of tablets, dragees, capsules, capsules, oral or injectable solutions or suspensions, suppositories, etc. dosed to allow daily administration of 1 to 1000 mg of active substance.

Claims (4)

R4 represents either a furan-2-yl group, a thien-2-yl or thien-3-yl group optionally substituted in the 5-position by a methyl group. R3 represents a hydrogen atom, and R2 represents a methyl group, R1 represents a methyl group, X represents a hydrogen or chlorine atom or a methyl or trifluoromethyl group,  in which

 1. Compound of general formula (I) 2. Process for the preparation of compounds according to claim 1, characterized in that an amide of general formula (II) is reacted.

 in which R 1 and R 4 are as defined in claim 1, with thionyl chloride, to obtain a chlorimine of general formula (III)

 that is reacted with an anhydride of general formula (IV)

 wherein X is as defined in claim 1 to obtain an isoquinolinone of the general formula (V)

 that it is treated with phosphorus oxychloride in the presence of N, N-dimethylformamide, to obtain, after hydrolysis, an aldehyde of general formula (VI)

 then the latter is treated with methyl (dimethoxyphosphinyl) acetate or with methyl (triphenylphosphoranylidene) acetate, to obtain an ester of general formula (VII)

 which is subjected to hydrogenation, to obtain an ester of general formula (VIII)

 and finally treating it with an amine of the general formula HNR2R3, wherein R2 and R3 are as defined in claim 1. 3. Medicinal product characterized in that it consists of a compound according to claim 1. 4. Pharmaceutical composition characterized in that it contains a compound according to claim 1, associated with an excipient.