WO1992009602A1 - Pyridobenzoindole derivatives, their preparation method, and pharmaceutical compositions containing same - Google Patents

Abstract

Novel pyridobenzoindole derivatives having general formula (I), where R is H or C1-2? alkyl, alk is linear or branched C2-4? alkylene, R1? is a hydrogen atom or a C1-2? alkyl radical, and R2? is a hydroxy or methoxy radical, and acid addition salts thereof, are useful as antitumoral agents.

Description

 PYRIDOBENZOINDELE DERIVATIVES, THEIR PREPARATION AND THE PHARMACEUTICAL COMPOSITIONS CONTAINING THEM

 The present invention relates to new pyridobenzoindole derivatives of general formula:

le cas échéant leurs hydrates, leurs sels d'addition avec les acides, leur préparation et les compositions pharmaceutiques qui les contiennent.

where appropriate their hydrates, their addition salts with acids, their preparation and the pharmaceutical compositions which contain them.

 In the European patent application 239,476 have been described γ-carboline derivatives of general formula:

in which R ₁ and R ₅ are inter alia hydrogen atoms, R ₂ can represent a hydrogen atom, a hydroxy or aleoyloxy radical, R ₃ and 3 ₁₄ are in particular alkyl radicals and n equal 2 to 4, which have anti-tumor activity.

 Furthermore, the pyridoindole derivatives unsubstituted in position -4 are described in the literature [J. Med. Chem., 31,

398 (1988)] as inactive compounds.

 The new pyridobenzoindole derivatives of general formula (I) in which:

R represents a hydrogen atom or an alkyl radical containing 1 or 2 carbon atoms, alk represents a straight or branched alkylene radical containing 2 to 4 carbon atoms, R ₁ represents a hydrogen atom or an alkyl radical containing 1 or 2 carbon atoms and

R ₂ represents a hydroxy or methoxy radical,

 have particularly interesting anti-tumor properties.

 According to the invention, the products of general formula (I) can be obtained from the chlorinated derivative of general formula:

dans laquelle R₁, est défini comme précédemment par action d'une aminé de formule générale :

in which R ₁ , is defined as above by the action of an amine of general formula:

H ₂ N - alk - K (R) ₂ (III) in which alk and R are defined as above, followed where appropriate by the transformation of the methoxylated product thus obtained, of general formula

dans laquelle R, R₁ et alk sont définis comme ci-dessus, en un dérivé hydroxy-9 pyrido[4,3-b]benzo[e]indole.

wherein R, R ₁ and alk are defined as above, to a 9-hydroxy pyrido [4,3-b] benzo [e] indole derivative.

 The reaction of the amine of general formula (III) with the chlorinated derivative of general formula (II) is carried out in the presence of an excess of the amine, preferably under nitrogen, optionally in an inert organic solvent, or without solvent, at a temperature between the reflux temperature of the reaction mixture and 250 ° C (autoclave).

 Éthylemethylation is carried out by any known method which does not alter the rest of the molecule.

The chlorinated derivative of general formula (II) for which R ₁ is a hydrogen atom can be prepared from the corresponding pyridone, of formula:

par action d'un agent de chloration.

by action of a chlorinating agent.

 It is generally carried out by means of a chlorinating agent chosen from phosphorus oxychloride or halogenated derivatives of phosphorus in the presence of a tertiary base (diethylaniline, dimethylaniline for example) in an organic solvent such as a nitrile (acetonitrile for example), at a temperature between 75 and 90 ° C. (reflux temperature of the reaction mixture).

Preferably, the reaction is carried out under nitrogen. The product of general formula (II) for which R ₁ is an alkyl radical may be obtained by alkylation of the product for which R ₁ is a hydrogen atom, for example by the action of the suitable halogen derivative, in the presence of potassium carbonate. The product of formula (IV) can be prepared from the hydrazone of formula:

par réaction de Fisher thermique, par analogie avec la méthode décrite par C.H. NGUYEN et E. BISAGNI, Tetrahedron, 42 (8), 2203 (1986).

by thermal Fisher reaction, by analogy with the method described by CH NGUYEN and E. BISAGNI, Tetrahedron, 42 (8), 2203 (1986).

 The hydrazone of formula (V) is prepared by condensation of hydrazino-4-1H-pyridone-2 with 6-methoxy-2-tetralone. The reaction is generally carried out in an organic solvent such as an alcohol (ethanol for example), at the reflux temperature of the reaction mixture.

 6-methoxy tetralone can be obtained according to the method described in J. Am. Chenu Soc, 82, 2573 (1960).

 Hydrazino-4-1H-pyridone-2 can be prepared according to the method described by C.H. NGUYEN and E. BISAGNI, Tetrahedron, 42 (8), 2203 (1986).

 The pyridobenzoindole derivatives of general formula (I) can be purified by crystallization or by chromatography.

 The new pyridobenzoindole derivatives according to the invention can be converted into addition salts with acids, by the action of an acid in an organic solvent. The salt precipitates, possibly after concentration of its solution, it is separated by filtration or decantation.

As pharmaceutically acceptable salts, mention may be made of addition salts with mineral acids such as hydrochlorides, hydrobromides, sulfates, nitrates, phosphates, or organic salts such as acetate, propionates, succinates, naleates, fumarates, methanesulfonates, p.toluenesulfonates, isethionates or derivatives of substitution of these compounds.

 The pyridobenzoindole derivatives according to the invention and their salts can exist in the form of hydrates, it is understood that these hydrated forms also come within the scope of the present invention.

 In addition, when the symbol alk is a branched alkylene radical, the pyridobenzoindole derivatives have isomeric forms. It is understood that these isomeric forms also come within the scope of the invention.

 The derivatives of general formula (I) are particularly advantageous as anti-tumor agents.

 Their anti-tumor properties have in particular been demonstrated in vitro at a concentration close to 15 to 1.5 μg / disc in the differential cytotoxicity test, according to the technique described by TH Corbett et al., Investigational new drug, 4, 207-220 (1986), and in vivo on grafted tumors of mice, more particularly on leukemia P388, the product studied was shown to be active at a dose of 20 mg / kg intraperitoneally.

 Their toxicity in mice expressed by their maximum tolerated dose is greater than 20 and 40 mg / kg intraperitoneally.

 Of particular interest for their anti-tumor activity are mainly the pyridobenzoindole derivatives of general formula (I), for which:

R is a methyl radical, alk is a straight or branched alkyl radical containing 2 to 4 carbon atoms, R ₁ is a hydrogen atom or a methyl radical, and R ₂ is a hydroxy radical, as well as their salts and the case where appropriate their hydrates.

 In particular [(dimethylamino-3) propyl] amino-1 hydroxy-9-5H-pyrido [4,3-b] benzol [e] indole, as well as its salts and its hydrated forms,

The following examples, given without limitation, illustrate the present invention. EXAMPLE 1

 [(3-dimethylamino) propyl] amino-1 methoxy-9 5H-pyrido [4,3-b] benzo [e] indole (free base) is released by excess ammonia from 1.8 g of corresponding bimaleate then extracted and dried.

The product thus obtained is placed in a 250 cm 3 reaction flask fitted with a stirrer under argon, with 66 cm ³ of hydrobromic acid (d = 1.47; 47%). This mixture, which has become homogeneous at boiling, is heated at reflux for 6 hours and concentrated to dryness under reduced pressure. The solid residue is dissolved in water (200 cm ³ ) made alkaline with 28% ammonia (20 cm ³ ) which is added drop by drop, to form a gummy precipitate.

To the resulting mixture, 200 cm ³ of ethyl acetate are added and the whole is stirred for one hour. The organic phase is decanted and the mother liquors saturated with sodium chloride are extracted twice with 100 cm ³ of ethyl acetate. The organic phase is dried over sodium sulfate, filtered, treated with 5 g of animal charcoal to discolor it, then filtered and concentrated to dryness.

The solid residue is taken up in 40 cm ³ of ethanol, and treated with 2.5 g of maleic acid in solution in 20 cm ³ of ethanol at the boil. [(Dimethylamino-3) propyl] amino-1 hydroxy-9 5H-pyrido [4,3-b] benzo [e] indole bimaleate hydrate with 0.25 H ₂ O (1.25 g, Yield = 70% ) is thus obtained, in the form of microcrystals; F = 195-200 ° C (decomposition).

 [(Dimethylamino-3) propyl] amino-1-methoxy-9 5H-pyrido [4,3-b] benzo [e] indole bimaleate can be obtained in the following manner:

Chloro-1 methoxy-9 5H-pyrido [4,3-b] benzo [e] indole (1.8 g) is placed in a 250 cm ³ flask containing 50 cm ³ (large excess) of 3-dimethylamino propylamine and heated to reflux (150 ° C), under nitrogen and with stirring, for 48 hours (total disappearance of the chlorinated derivative, checked on silica plate).

The excess diamine is evaporated in a water bath under reduced pressure and the residue is taken up in water and then made alkaline with ammonia. The solid formed is filtered, washed with water, taken up in methylene chloride and washed with 150 cm ³ of water and 10 cm ³ of ammonia.

 After chromatography on alumina and salification with maleic acid under the usual conditions, 2.56 g (R = 69%) of [(dimethylamino-3) propyl] bimaleate-9-methoxy-5H-pyrido [4H] bimaleate are obtained. , 3-b] benzo [e] indole expected, melting at 204-206 ° C with decomposition.

 Chloro-1 methoxy-9 5H-pyrido [4,3-b] benzo [e] indole can be obtained in the following way:

4 g of methoxy-9-2H, 5H-pyrido [4,3-b] are introduced into a 1 liter flask fitted with a magnetic stirrer, an addition funnel, a condenser and kept under nitrogen. ] benzo [e] indolone-1 taken up in boiling ethanol and carefully dried, 14.1 g (4 equivalents) of benzyl triethylammonium chloride, 3.6 g (4 equivalents) of acetamide, 40 cm ³ of acetonitrile and 9.12 g (4 equivalents) of freshly rectified aniethylaniline.

70 cm ³ of rectified phosphorus oxychloride are gradually added through the bulb, and an exothermic reaction is observed. The mixture is heated at reflux for 20 hours, while after having passed through a homogeneous phase, a precipitate appears. Concentration to dryness, in a water bath heated to 70 ° C and under reduced pressure (15 mm Hg).

To the residue obtained, 300 cm ³ of ice water are added and the mixture, well stirred for 2 hours, is heated at 70 ° C for 20 minutes. It is made up to 600 cm ³ with water, heated to boiling for 5 minutes and allowed to cool to room temperature with stirring. After cooling, a yellow precipitate is obtained, which is filtered cold and then washed with cold water. This precipitate is taken up in 500 cm ³ of distilled water and the mixture is basified, cold, with ammonia. The whole is left stirring for 1 night and the precipitate is filtered and washed with water. It is recrystallized from alcohol, giving 2.6 g of yellow needles of 1-chloro-9-methoxy 5H-pyrido [4,3-b] benzo [e] indole; M> 260 ° C;

Yield 60%.

 Methoxy-9-2H, 5H-pyrido [4,3-b] benzo [e] indolone-1 can be obtained in the following way:

In a 1 liter three-necked flask, the N- (1H-pyriάone-2) is mixed yl) -4 N '- (6-methoxy-1,2,3,4-naphthalenylidene) -2 hydrazine (18 g) and diphenyl ether (700 cm ³ ) then heated at reflux for 30 minutes, maintaining the together under good agitation and under nitrogen. The mixture becomes homogeneous and discolours. The heating is interrupted to allow it to cool to 200 ° C. and to check that the starting compound is completely transformed: Merck silica plate, eluent methylene chloride-ethanol 8/2 by volume, (rf hydrazone = 0.5, rf intermediate compound = 0 , 8). Continuing to stir, the 10% palladium-on-charcoal (4 g) suspended in diphenyl ether (20 cm ³ ) is added gradually, with caution (foams and release of hydrogen), and the new mixture is heated at reflux for 30 minutes (disappearance of the intermediate compound) (rf = 0.35 silica-pure ethyl acetate; expected product: rf = 0.58). The mixture is then taken up in 600 cm ³ of boiling ethoxy-2 ethanol, filtered to remove the palladium-on-carbon and concentrated to dryness. The solid obtained is added with 70 cm ³ of absolute ethanol, stirred for 18 hours, filtered and dried to give 11 g (R = 68%) of yellowish microcrystals (F> 260ºC) corresponding to methoxy-9-2H, 5H -pyrido [4,3-b] benzo [e] indolone-1.

 N- (1H-pyridone-2 yl) -4 N '- (6-methoxy-1,2,3,4 tetrahydro naphthalenylidene) -2 hydrazine can be obtained in the following way:

The mixture of hydrazino-4-1H-pyridone-2 (9.8 g) in absolute ethanol (150 cm ³ ) is heated to reflux. To the solution, which has become homogeneous at boiling, 6-methoxy-tetralone-2 (16.6 g) is added and the mixture, which has become rapidly heterogeneous, is heated to reflux for 2 hours (disappearance of the hydrazine controlled by CCH on Merck silica plate, eluting methylene chloride-ethanol 1/1 by volume). The precipitate formed is filtered, taken up in ethanol in which the hydrazone is poorly soluble and then filtered cold to give (19.3 g (83%)) of yellow microcrystals of N- (1H-pyridone-2 yl) - 4 N '- (6-methoxy-1,2,3,4-naphthalenylidene tetrahydro) -2 hydrazine, mp 220-225 ° C, with decomposition.

Hydrazino-4-1H-pyridone-2 can be obtained according to the method described by E. Bisagni and CH Nguyen, Tetrahedron, 42, 2303 (1986). The present invention also relates to pharmaceutical compositions which contain as active product at least one product of general formula (I) in the pure state (in free form or in salt form) or in combination with one or more pharmaceutically acceptable adjuvants. These compositions can be used parenterally.

 The compositions for parenteral administration can be sterile aqueous or non-aqueous solutions, suspensions or emulsions. As solvent or vehicle, propyleneglycol, a polyethylene glycol, vegetable oils, in particular olive oil and injectable organic esters, for example ethyl oleate, can be used. These compositions can also contain adjuvants, in particular wetting agents, emulsifiers or dispersants. Sterilization can be done in several ways, for example using a bacteriological filter, by incorporating sterilizing agents into the composition, by irradiation or by heating. They can also be prepared in the form of sterile solid compositions which will be dissolved at the time of use in sterile water or any other sterile injectable medium.

 In human therapy, the medicaments according to the present invention are particularly useful in the treatment of digestive cancers, of lung cancers, of cancers of the testes or of the ovaries as well as in the treatments of cancers of the head and of the neck.

 In general, the doctor will determine the dosage he considers most appropriate based on age, weight and all other factors specific to the subject to be treated.

The preferred mode of administration is the intravenous route. As an indication, the medicaments according to the invention can be administered to humans at a rate of 30 to 200 mg / m ² per treatment, by intravenous route. The following example, given without limitation, illustrates a composition according to the present invention:

EXAMPLE:

 A solution containing 8.54 g of [(dimethylamino-3) propyl] 1-amino-hydroxy-95H-pyrido [4,3-b] benzo bimaleate is prepared.

[e] hydrated indole (0.5 H ₂ O), by dissolving this product in pyrogen-free physiological solution in sufficient quantity to obtain

100 cm ³ .

The solution obtained is aseptically distributed in ampoules, at the rate of 2 cm ³ per ampoule. The ampoules are sealed and each contain 100 mg of [(dimethylamino-3) propyl] amino-1 hydroxy-9 5H-pyrido [4,3-b] benzo [e] indole.

Claims

1 - A new pyridobenzoindole derivative of general formula

in which  R represents a hydrogen atom or an alkyl radical containing 1 or 2 carbon atoms, alk represents a straight or branched alkyl radical containing 2 to 4 carbon atoms, R ₁ represents a hydrogen atom or an alkyl radical containing 1 or 2 carbon atoms, and R ₂ represents a hydroxy or methoxy radical, as well as its addition salts with acids, and where appropriate its hydrates and its isomeric forms and their mixtures. 2 - A new pyridobenzoindole derivative according to claim 1 for which:  R is a methyl radical, alk is a straight or branched alkyl radical containing 2 to 4 carbon atoms, R ₁ is a hydrogen atom or a methyl radical, and R ₂ is hydroxy radical as well as its salts and, where appropriate, its hydrates and its isomeric forms and their mixtures. 3 - [(3-dimethylamino) propyl] amino-1 hydroxy-9-5H-pyrido [4,3-b] benzo [e] indole, as well as its salts and hydrated forms. 4 - A process for the preparation of a pyridobenzoindole derivative according to claim 1, characterized in that an amine of general formula is made to act: H ₂ N - alk - N (R) ₂ in which alk and R are defined as in claim 1, on a chlorinated derivative of formula:

in which R ₁ is defined as in claim 1, then transforms, where appropriate, the methoxylated product obtained, of general formula:

wherein R ₁ is defined as in claim 1, in one derivative 9-hydroxy pyrido [4,3-b] benzo [e] indole and optionally transforms the product obtained into an addition salt with an acid. 5 - Pharmaceutical composition characterized in that it contains a product according to claim 1 in the pure state or in the form of an association with any compatible and pharmaceutically acceptable diluent or adjuvant.