HK1034073B - Nitrogen-containing tetracyclic compounds - Google Patents

Description

Nitrogen-containing tetracyclic compounds

Technical Field

The present invention relates to compounds having a high affinity for mitochondrially diazepam binding inhibitor receptors (MDR for short).

Background

Benzodiazepine (BZ) receptor, a site of action of sedative-hypnotic drugs, can be classified as being present in GABA_AThe central benzodiazepine * receptor (CBR) on the receptor/chloride channel complex and the MDR 2 species present in the central nervous system (glial cells) or adrenal gland (clin. neuropharmacol., 16, 401. 417, 1993). CBR agonists represented by diazepam are widely used as sedative hypnotic agents, but since CBR agonists act directly on GABA_AThe receptor/chloride channel complex exhibits sedative-hypnotic activity and also exhibits side effects such as excessive sedation or mental dependence. On the other hand, MDR agonists act indirectly on GABA by synthesis of neuroactive steroids (endogenous sedative hypnotic substances), neurocholesterol_AThe receptor/chloride channel, while exhibiting sedative-hypnotic effects, also exhibits side effects such as excessive sedation or neuro-dependence (J.Pharmacol. exp. Ther., 267, 462-471, 1993; ibid., 265, 649-656, 1993).

Therefore, it is desired to develop an MDR agonist as a therapeutic agent for symptoms (obsessive-compulsive disorder, panic disorder) which have not sufficiently exhibited a therapeutic effect on the conventional BZ-series, and a sedative hypnotic agent which alleviates the side effects of the conventional BZ-series.

In addition, compounds acting on MDR are due to GABA_AThe receptor action may become a therapeutic agent for insomnia, epilepsy, dyskinesia with muscular stiffness, eating disorder, circulatory disorder, learning disorder and drug dependence (Progress in neurobiology, 38, 379-. Furthermore, from the physiological function of MDR, it is possible to be a therapeutic agent for cancer (Biochimica et BIOphysica Acta, 1241, 453-470, 1995), lipid metabolism disorder (Eur. J. Pharmacol., 294, 601-607, 1995), schizophrenia (Neuropharmacology, 35, 1075-1079, 1996), cerebral infarction (J. Neurosci., 15, 5263-5274, 1995), ADIS (extracts of the after international conference AIDS, P458, 1989), Alzheimer's disease (Alzheimer's disease. Assoc. Disod.2, 331-336, 1988) or Huntington's chorea (Brain Res., 248, 396-401, 1982).

As compounds having affinity for MDR, indole compounds disclosed in Japanese patent application laid-open No. 6-501030 are mentioned.

Description of the invention

The present inventors have made intensive studies on a compound having a higher affinity for MDR, and as a result, have found that a specific nitrogen-containing tetracyclic compound achieves the object, thereby completing the present invention. As described above, indole compounds are known to have an affinity for MDR, but no nitrogen-containing tetracyclic compounds have been reported to have an affinity for MDR.

That is, the present invention is a nitrogen-containing tetracyclic compound represented by the formula [ I ] or a pharmaceutically acceptable salt thereof.

[ in the formula, Y¹-Y²-Y³Represents N-C-N or formula C-NR³(in the formula, R³Represents a hydrogen atom, C_1-5Alkyl or C_2-10Nitrogen-containing alkyl group of (b). Y is⁴Denotes S, SO₂、CH₂Or NR⁴(in the formula, R⁴Is represented by C_1-5Alkanoyl or C_1-5Alkyl) of (a). R¹And R²The same or different represent a hydrogen atom, C_1-10Alkyl of (C)_3-15Alkoxyalkyl or C_3-15Or R is alkyl amino alkyl, or¹And R²Together with the adjacent nitrogen atom, form a cyclic amino group. X¹And X²The same or different represent a hydrogen atom, C_1-5Alkyl of (C)_1-5Alkoxy or halogen atom of (2), n represents 0, 1 or 2)

In the present invention, R³、R⁴、X¹、X²C of (A)_1-5The alkyl group represents a linear, branched or cyclic alkyl group, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, cyclopropylmethyl, pentyl, isopentyl and the like. R³C of (A)_2-10Nitrogen-containing alkyl groups such as methylaminopropyl, dimethylaminoethyl, pyrrolidinylethyl, 4-methylpiperazinylethyl and the like. R⁴C of (A)_1-5Alkanoyl groups such as formyl, acetyl, propionyl and the like. R¹、R²C of (A)_1-10The alkyl group represents a linear, branched or cyclic alkyl group, and examples thereof include a methyl group, an ethyl group, a propyl group, an isopropyl group, a cyclopropyl group, a butyl group, an isobutyl group, a cyclobutyl group, a cyclopropylmethyl group, a pentyl group, an isopentyl group, a cyclopentyl group, a cyclobutylmethyl group, a 1-ethylpropyl group, a hexyl group, an isohexyl group, a cyclohexyl group, a cyclopentylmethyl group, a 1-ethylbutyl group, a heptyl group, an isoheptyl group, a cyclohexylmethyl group, an octyl group, a nonyl group, and a decyl group. R¹、R²C of (A)_3-5Alkoxyalkyl represents a straight-chain, branched or cyclic C_1-13alkoxy-C_2-14Alkyl radicals, e.g. methylOxyethyl, methoxypropyl, methoxybutyl, ethoxyethyl, ethoxypropyl, ethoxybutyl, ethoxypentyl, ethoxyhexyl, ethoxyheptyl, propoxyethyl, propoxypropyl, propoxybutyl, isopropoxyethyl, cyclopropylmethoxyethyl and the like. R¹、R²C of (A)_3-15Alkylaminoalkyl represents a straight, branched or cyclic C_1-13alkylamino-C_2-14Alkyl groups such as methylaminoethyl, dimethylaminoethyl, methylaminopropyl, dimethylaminopropyl, methylaminobutyl, ethylaminoethyl, ethylaminopropyl, ethylaminobutyl, ethylaminopentyl, ethylaminohexyl, ethylaminoheptyl, ethylaminooctyl, propylaminoethyl, propylaminopropyl, propylaminobutyl, isopropylaminoethyl, cyclopropylmethylaminoethyl, pyrrolidinylethyl and the like. R¹、R²And a cyclic amino group formed adjacent thereto by a nitrogen atom such as pyrrolidinyl, piperidinyl, homopiperidinyl, morpholinyl, piperazinyl, N-methylpiperazinyl, 3, 5-dimethylpiperazinyl and the like. X¹、X²C of (A)_1-5The alkoxy group represents a linear, branched or cyclic alkoxy group, and examples thereof include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a butoxy group, an isobutoxy group, a cyclopropylmethoxy group, a pentyloxy group, and an isopentyloxy group. X¹、X²The halogen atom of (a) represents a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

The pharmaceutically acceptable salts in the present invention include salts with inorganic acids such as sulfuric acid, hydrochloric acid, and phosphoric acid, and salts with organic acids such as acetic acid, oxalic acid, lactic acid, tartaric acid, fumaric acid, maleic acid, methanesulfonic acid, and benzenesulfonic acid.

The compound of the formula [ I ] can be produced by the following conventional production methods 1 to 6 (in the following reaction scheme, Y¹、Y²、Y³、Y⁴、R¹、R²、X¹、X²And n is the same as above, R⁵Represents a hydrogen atom or C_1-5Alkyl of R⁶Is represented by C_1-5Alkyl or C_2-10Nitrogen-containing alkyl of R⁷And R⁸Is the same as orIn the different, represents C_1-5Alkyl or benzyl of, X³Represents a chlorine atom, a bromine atom or an iodine atom, X⁴Represents a fluorine atom, a chlorine atom, a bromine atom or an iodine atom, and Boc represents a tert-butoxycarbonyl group).

[ conventional preparation method 1 ]

Step A: a tetracyclic indole derivative (3) is produced by a Fischer indole synthesis method using a keto acid derivative (1) and a phenylhydrazine derivative (2). R of tetracyclic indole derivative (3)⁵Is C_1-5When the alkyl group is substituted, the ester thereof may be hydrolyzed by a common base or acid to derive the carboxylic acid derivative (R)⁵＝H)。

And B: the compound (5) of the present invention can be synthesized from the tetracyclic indole derivative (3) via an acid halide, a mixed acid anhydride or the like.

The acid halide here represents an acid chloride, an acid bromide or the like, and can be reacted with the tetracycloindole derivative (3) (R) in an inert solvent by using a halogenating agent such as thionyl chloride, thionyl bromide, oxalyl chloride, carbon tetrachloride-triphenylphosphine, carbon tetrabromide-triphenylphosphine or the like⁵H) reaction. The inert solvent is ethers such as tetrahydrofuran and the like; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as chloroform and methylene chloride, acetonitrile, and N, N-dimethylformamide.

The mixed acid anhydride represents a carboxylic acid derivative (3) (R)⁵H) with a carbonate or a carboxylic acid, and the like, can be obtained by reacting a halogenated carbonate such as ethyl chlorocarbonate or isobutyl chlorocarbonate, or a carboxylic acid such as acetic acid, propionic acid, benzoic acid or naphthoic acid in an inert solvent in the presence of an organic base such as triethylamine, diisopropylethylamine, N-methylmorpholine or pyridine, or an inorganic base such as sodium hydroxide. The inert solvent is ethers such as tetrahydrofuran and the like; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as chloroform and methylene chloride, acetonitrile, and N, N-dimethylformamide.

The compound (5) of the present invention can also be obtained by reacting the tetracyclic indole derivative (3) with a condensing agent and an amine (4) in an inert solvent.

The condensing agent herein means an amidation agent generally used, and examples thereof include diphenylphosphoramide, diethyl cyanophosphate, carbonyldiimidazole, N '-dicyclohexylcarbodiimide, N-ethyl-N' -dimethylaminopropylcarbodiimide hydrochloride and the like. Inert solvents such as ethers like 1, 2-dimethoxyethane, tetrahydrofuran, etc.; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as chloroform and methylene chloride; acetonitrile, N-dimethylformamide, and the like. In addition, if necessary, N-hydroxysuccinimide, 1-hydroxybenzotriazole, 3-hydroxy-4-oxo-3, 4-dihydro-1, 2, 3-benzotriazine, or the like may be added as an activator in the present reaction.

[ conventional preparation method 2 ]

The compound (5) of the present invention can also be obtained by amidating a keto acid (6) in step B and then reacting the resulting ketoamide derivative (7) under the Fischer indole synthesis conditions of step A.

[ conventional preparation method 3 ]

And C: compound (9) of the present invention can be obtained by reacting tetracyclic compound (5) with halogenated compound (8) in an inert solvent in the presence or absence of a phase transfer catalyst together with a base.

Inert solvents herein such as alcohols such as methanol and ethanol; ethers such as 1, 2-dimethoxyethane and tetrahydrofuran; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as chloroform and methylene chloride; acetonitrile, N-dimethylformamide, and the like. A phase transfer catalyst such as a quaternary ammonium salt such as benzyltriethylammonium bromide, tetrabutylammonium bromide and the like; crown ethers such as 18-crown-6-ether, and the like. Bases such as inorganic bases including potassium carbonate, sodium hydroxide, sodium hydride, and metallic sodium; alcoholates such as potassium tert-butoxide and sodium ethoxide.

[ conventional preparation method 4 ]

Step D: the benzylmalonic acid derivative (12) can be produced by reacting a 2-cyanobenzyl halide (10) with a 2-aminomalonate diester derivative (11) in an inert solvent in the presence or absence of a phase transfer catalyst together with a base.

Inert solvents such as alcohols such as methanol and ethanol; ethers such as 1, 2-dimethoxyethane and tetrahydrofuran; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as chloroform and methylene chloride; acetonitrile, N-dimethylformamide, and the like. A phase transfer catalyst such as a quaternary ammonium salt such as benzyltriethylammonium bromide, tetrabutylammonium bromide and the like; crown ethers such as 18-crown-6-ether, and the like. Bases such as inorganic bases including potassium carbonate, sodium hydroxide, sodium hydride, and metallic sodium; alcoholates such as potassium tert-butoxide and sodium ethoxide.

Step E: the phenylalanine derivative (13) can be obtained by hydrolyzing an ester with a base or an acid in an inert solvent and then decarboxylating the ester. Inert solvents such as alcohols such as methanol and ethanol; ethers such as 1, 2-dimethoxyethane and tetrahydrofuran; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as chloroform and methylene chloride; acetonitrile, N-dimethylformamide, water or a mixed solvent thereof. Bases such as inorganic bases such as potassium carbonate, sodium carbonate, potassium hydroxide, sodium hydroxide, etc., acids such as hydrochloric acid, sulfuric acid, phosphoric acid, etc.

The phenylalanine derivative (13) can be obtained by reacting in step B, and then deprotecting it by treatment with an organic acid such as trifluoroacetic acid or formic acid, or an inorganic acid such as hydrogen chloride, hydrochloric acid, hydrobromic acid, or sulfuric acid to give an amide compound (14).

Step F: the aniline derivative (16) can be derived by reacting the amide compound (14) with the nitrobenzene derivative (15) in an inert solvent in the presence or absence of a base.

Examples of the base include inorganic bases such as potassium carbonate, sodium hydroxide, sodium hydride and metallic sodium, alcoholates such as potassium tert-butoxide and sodium ethoxide, and organic bases such as triethylamine, diisopropylethylamine and pyridine. Inert solvents such as alcohols such as methanol and ethanol; ethers such as 1, 2-dimethoxyethane and tetrahydrofuran; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as chloroform and methylene chloride; acetonitrile, N-dimethylformamide, and the like.

Step G: the compound (17) of the present invention can be obtained by reducing the nitro group of the aniline derivative (16) in an inert solvent and then subjecting the resultant to an acid treatment in an inert solvent.

The reduction herein refers to hydrogenation using platinum dioxide, palladium, or the like, or metal reduction using a metal such as tin, iron, zinc, or a metal salt such as stannous chloride under acidic, neutral, or alkaline conditions. In this case, an inert solvent such as alcohols such as methanol and ethanol; ethers such as diethyl ether and tetrahydrofuran; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as dichloromethane and chloroform; organic carboxylic acids such as acetonitrile, N-dimethylformamide, and acetic acid, water, or a mixed solvent thereof. The acid treatment in the inert solvent is carried out by reacting an alcohol such as methanol or ethanol with an acid such as hydrogen chloride, hydrogen bromide, sulfuric acid or trifluoroacetic acid in a single solvent or in a mixed solvent of an alcohol such as ethers such as diethyl ether or tetrahydrofuran, hydrocarbons such as toluene or benzene, halogenated hydrocarbon solvents such as methylene chloride or chloroform, or a mixed solvent of an alcohol with N, N-dimethylformamide.

[ conventional preparation method 5 ]

Step H: racemic or optically active sulfoxide derivatives (19) or sulfone derivatives (20) can be obtained by treating tetracyclic compounds (18) containing a sulfur atom with an oxidizing agent in an inert solvent.

Inert solvents such as alcohols such as methanol and ethanol; ethers such as diethyl ether and tetrahydrofuran; hydrocarbons such as toluene and benzene; halogenated hydrocarbon solvents such as dichloromethane and chloroform; organic carboxylic acids such as acetonitrile, N-dimethylformamide, and acetic acid, water, or a mixed solvent thereof. Oxidizing agents such as percarboxylic acids (e.g., m-chloroperbenzoic acid, peracetic acid, etc.), hydrogen peroxide, potassium peroxide (OXONE, 2 KSO)₅·KHSO₄·K₂SO₄) And the like.

[ conventional preparation method 6 ]

Step I: the optically active form (22) can also be prepared by resolution of the racemate (21) of the compound of the present invention by HPLC using a chiral stationary phase.

The chiral stationary phase is cellulose ester, cellulose carbamate, amylose carbamate, crown ether, polymethacrylate and other derivatives.

Step J: the racemic carboxylic acid derivative (23) can be resolved into the optically active form (24) by forming a salt with a chiral amine.

The chiral amine is (+) or (-) -1-phenylethylamine, (+) or (-) -2-amino-1-butanol, (+) or (-) -aminopropanol (アラニノ - ル), strychnine, cinchonidine, cinchonine, quinine, quinidine, dehydroabietylamine, etc.

The optically active substance (22) can be obtained by amidating the optically active substance (24) in step B.

[ conventional preparation method 7 ]

When R is the compound (21) or (22) of the present invention shown in general preparation Process 6¹、R²When either or both of them contain a nitrogen atom protected with an acyl group, an alkoxycarbonyl group or the like, these protecting groups can be removed by an acid or a base to obtain the compound of the present invention.

Here, the acid means trifluoroacetic acid, formic acid, hydrogen chloride, hydrogen bromide, hydrochloric acid, hydrobromic acid, etc., and the base means potassium carbonate, sodium carbonate, potassium hydroxide, sodium hydroxide, lithium hydroxide, barium hydroxide, etc.

Industrial applicability

The compounds of the invention have a high affinity for MDR. Therefore, it is useful as a therapeutic or prophylactic agent for restlessness or a disease related thereto, depression, epilepsy, sleep disorders, cognitive and learning disorders, central diseases such as schizophrenia, dyskinesia associated with muscular stiffness, eating disorders, circulatory disorders, drug dependence, cancer, lipid metabolism disorders, cerebral infarction, AIDS, Alzheimer's disease or Huntington's chorea.

Best mode for carrying out the invention

The present invention will be specifically described below with reference to examples and experimental examples.

Example 1

Preparation of N-2- (propylamino) ethyl-N-hexyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide

(1) To 100ml of ethanol solution of 22.54g of (4-oxo-thiochroman-2-yl) -carboxylic acid and 10.7ml of phenylhydrazine was added 15ml of sulfuric acid, and the mixture was refluxed for 5 hours. The reaction mixture was cooled to room temperature, poured into 500ml of ice water, and extracted with diethyl ether. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, the filtrate was concentrated under reduced pressure, and the residue was recrystallized from ethanol-hexane to give 21.85g of ethyl 6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxylate.

(2) 19.46g of potassium hydroxide was dissolved in 40ml of water, and the solution was added to 100ml of an ethanol solution containing 21.63g of ethyl 6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxylate. After heating and refluxing for 2 hours, concentrated hydrochloric acid was added dropwise to the reaction solution, the pH was adjusted to 3, and the mixture was extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, the filtrate was concentrated under reduced pressure, and the residue was recrystallized from ethanol-hexane to give 19.94g of 6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxylic acid.

m.p.141.5～142.5℃

(3) To a solution of 844mg of 6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxylic acid and 1.66g of N-2- (N-t-butoxycarbonylpropylamino) ethyl-hexylamine in 44ml of methylene chloride were added 552mg of 1-hydroxybenzotriazole monohydrate and 863mg of N-ethyl-N' -dimethylaminopropyl carbodiimide hydrochloride, and the mixture was stirred at room temperature for 1 night. The reaction mixture was concentrated under reduced pressure, and then dissolved in ethyl acetate, washed with water, a 5% aqueous sodium hydrogen sulfate solution, a saturated aqueous sodium bicarbonate solution and a saturated common salt solution, dried over anhydrous sodium sulfate, filtered to remove a drying agent, the filtrate was concentrated under reduced pressure, and the residue was treated with flash chromatography (silica gel: Chromatrex NHDM 1020 (fuji Devisone chemical corporation), a developing solvent: hexane-ethyl acetate ═ 2: 1 to 3: 2) to obtain 1.35g of amorphous N-2- (N-t-butoxycarbonylpropylamino) ethyl-N-hexyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide.

(4) In 4.2ml of 99% formic acid, 600mg of N-2- (N-tert-butoxycarbonylpropylamino) ethyl-N-hexyl-6, 11-dihydro-5-thioxo-11-aza-benzo [ a ] fluorene-6-carboxamide was stirred for 5 hours. The reaction mixture was concentrated under reduced pressure, and then dissolved in ethyl acetate, washed with a saturated aqueous sodium bicarbonate solution and a saturated common salt solution, dried over anhydrous sodium sulfate, and then the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure, and the residue was recrystallized from ethyl acetate to obtain 406mg of N-2- (propylamino) ethyl-N-hexyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide.

The structures and physical data of this compound and the compound obtained by the same are shown in table 1.

Example 2

Preparation of N, N-dihexyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide

(1) To a solution of 20.00g of (4-oxo-thiochroman-2-yl) -carboxylic acid in 200ml of benzene was added 14.0ml of thionyl chloride, and the mixture was refluxed for 3 hours. The reaction mixture was concentrated under reduced pressure, and 100ml of a dichloromethane solution of the residue was added dropwise to 200ml of a dichloromethane solution of 24.6ml of dihexylamine and 20.0ml of triethylamine under ice-water bath conditions with stirring. After stirring at room temperature for 1 night, the reaction mixture was concentrated under reduced pressure, ethyl acetate was added to the residue, which was washed with water, 1N hydrochloric acid, a saturated aqueous sodium bicarbonate solution, and a saturated saline solution, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography (silica gel: wacogel c200 (manufactured by wako pure chemical industries, Ltd.) and a developing solvent: hexane-ethyl acetate 5: 1 to 3: 1), and then recrystallized from hexane to obtain 29.22g of N, N-dihexyl- (4-oxo-thiochroman-2-yl) -carboxamide.

(2) After stirring 1.00g of N, N-dihexyl- (4-oxo-thiochroman-2-yl) -carboxamide and 0.26ml of phenylhydrazine at 100 ℃ for 30 minutes, the reaction mixture was dried under reduced pressure at 50 ℃ for 30 minutes. To the residue was added 1.44g of anhydrous zinc chloride, and the mixture was stirred at 170 ℃ for 5 minutes and cooled to room temperature. To the reaction mixture was added ice water, and the mixture was extracted with ethyl acetate, washed with 1N hydrochloric acid, a saturated aqueous sodium bicarbonate solution and a saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure. The residue was recrystallized from ethyl acetate-hexane to give 0.79g of N, N-dihexyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide.

(3) The racemic N, N-dihexyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide was resolved by high performance liquid chromatography (chiralpak AD (Daicel Co., Ltd.), 2. phi. times.25 cm, mobile phase: hexane-ethanol 3: 7, flow rate: 5 ml/min).

(-) -N, N-dihexyl-6, 11-dihydro-5-thioxo-11-aza-benzo [ a ] fluorene-6-carboxamide

[α]_D ²⁶-25.9(c ═ 0.180, EtOH), retention time: and 20 min.

(+) -N, N-dihexyl-6, 11-dihydro-5-thioxo-11-aza-benzo [ a ] fluorene-6-carboxamide

[α]_D ²⁶+25.9(c ═ 0.207, EtOH), retention time: and (4) 37 min.

The structures and physical data of the compound and the compound obtained by the same are shown in tables 1 and 2.

Example 3

Preparation of N, N-dihexyl-6, 11-dihydro-11-methyl-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide

To a solution of 200mg of N, N-dihexyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide in 10ml of N, N-dimethylformamide was added 21mg of 60% sodium hydride/oil, and the mixture was stirred at room temperature for 1 hour. To the solution was added 33. mu.l of methyl iodide, and the mixture was stirred at room temperature for 5 hours. To the reaction solution was added ethyl acetate, which was washed with water, 1N hydrochloric acid, a saturated aqueous sodium bicarbonate solution, and a saturated common salt solution, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure. The residue was recrystallized from hexane to give 155mg of N, N-dihexyl-6, 11-dihydro-11-methyl-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide.

The structures and physical data of this compound and the compound obtained by the same are shown in table 2.

Example 4

N, N-dihexyl-5, 6-dihydro-benzo [ 4, 5 ] imidazo [ 2, 1-a ] isoquinoline-6-carboxamide

(1) 0.49g of sodium was dissolved in 20ml of ethanol, and a solution of 5.90g of diethyl 2-N-t-butoxycarbonylaminomalonate in 10ml of ethanol was added thereto at room temperature with stirring. After stirring for 20 minutes, a 10ml solution of 2-cyanobenzylbromide (4.00 g) in ethanol was added to the reaction mixture, and the mixture was stirred at room temperature for 10 minutes and then under reflux for 3.5 hours. The reaction mixture was concentrated under reduced pressure, ethyl acetate was added to the residue, which was washed with water, a 5% aqueous solution of potassium hydrogensulfate, a saturated aqueous solution of sodium hydrogencarbonate and saturated brine, dried over anhydrous sodium sulfate, and then the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography (silica gel: Wacogel C200 (Wako pure chemical industries, Ltd.), developing solvent: hexane-ethyl acetate 5: 1) to give 7.86g of diethyl 2- (2-cyanophenylmethyl) -2-N-tert-butoxycarbonylaminomalonate as an oil.

(2) To 1.17g of diethyl 2- (2-cyanophenylmethyl) -2-N-tert-butoxycarbonylaminomalonate were added 20ml of ethanol and 0.36g of aqueous sodium hydroxide solution (0.5 ml of sodium hydroxide/water), and the mixture was stirred under reflux for 2 hours. The reaction mixture was concentrated under reduced pressure, and a 5% aqueous solution of potassium hydrogensulfate was added to the residue to conduct extraction with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to give 0.81g of crude N-tert-butoxycarbonyl- (2-cyanophenyl) alanine as an oil. The compound was used in the next step without purification.

(3) 0.81g of crude N-t-butoxycarbonyl- (2-cyanophenyl) alanine and 0.67g of dihexylamine were dissolved in 8ml of N, N-dimethylformamide, and thereto were added 0.55g of 1-hydroxybenzotriazole monohydrate and 0.69g of N-ethyl-N' -dimethylaminopropyl carbodiimide hydrochloride, followed by stirring at room temperature for 1 night. The reaction mixture was poured into water, extracted with ethyl acetate, washed with a 5% aqueous potassium hydrogensulfate solution, a saturated aqueous sodium hydrogencarbonate solution and a saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography (silica gel: Wacogel C200 (manufactured by Wacogel Co., Ltd.), and a developing solvent: hexane-ethyl acetate 4: 1) to give 1.01g of N-tert-butoxycarbonyl- (2-cyanophenyl) alanine dihexylamide.

(4) 0.98g of N-tert-butoxycarbonyl- (2-cyanophenyl) alanine dihexylamide was dissolved in 1.7ml of methylene chloride, and 1.7ml of trifluoroacetic acid was added thereto, followed by stirring at room temperature for 1.5 hours. The reaction mixture was concentrated under reduced pressure, and a saturated aqueous sodium hydrogencarbonate solution was added to the residue to conduct extraction with methylene chloride, and the residue was washed with saturated brine, dried over anhydrous sodium sulfate, and then the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain 0.76g of crude (2-cyanophenyl) alanine dihexylamide. The compound was used in the next step without purification.

(5) 0.76g of crude (2-cyanophenyl) alanine dihexylamide, 0.30g of 2-fluoronitrobenzene, and 0.36g of anhydrous potassium carbonate were refluxed in 8ml of N, N-dimethylformamide for 2.5 hours. The reaction mixture was poured into water, extracted with ethyl acetate, washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography (silica gel: Wacogel C200 (manufactured by Wacogel Co., Ltd.), and a developing solvent: hexane-ethyl acetate 5: 1) to give 0.42g of N- (2-nitrophenyl) - (2-cyanophenyl) alanine dihexylamide.

(6) 95mg of N- (2-nitrophenyl) - (2-cyanophenyl) alanine dihexylamide and 10mg of platinum dioxide were stirred in 3ml of methanol under a hydrogen atmosphere for 2 hours. After insoluble matter was removed by filtration using a Celite plate, the filtrate was concentrated under reduced pressure.

The residue was dissolved in 5ml of ethanol, and hydrogen chloride gas was introduced thereinto to obtain a saturated solution. The reaction mixture was stirred for 4 hours, poured into a saturated aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography (silica gel: Wacogel C200 (Wacogel Co., Ltd.), developing solvent: hexane-ethyl acetate 2: 1) and then recrystallized from ethyl acetate to give 24mg of N, N-dihexyl-5, 6-dihydro-benzo [ 4, 5 ] imidazo [ 2, 1-a ] isoquinoline-6-carboxamide.

The structures and physical data of this compound and the compound obtained by the same are shown in table 3.

Example 5

Preparation of N, N-dipropyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide 5, 5-dioxide

Under the ice-water bath condition, a solution of 710mg of m-chloroperbenzoic acid (containing 70% or more) in 30ml of dichloromethane was added dropwise to a solution of 500mg of N, N-dipropyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide in 30ml of dichloromethane over 20 minutes under stirring. After stirring at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure, the residue was dissolved in ethyl acetate, washed with a saturated aqueous sodium bicarbonate solution and a saturated brine, dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography (silica gel: Wacogel C200 (manufactured by Wacogel corporation), and a developing solvent: hexane-ethyl acetate 3: 1 to 1: 1), and then recrystallized from ethyl acetate-hexane to obtain 230mg of N, N-dipropyl-6, 11-dihydro-5-thio-11-aza-benzo [ a ] fluorene-6-carboxamide 5, 5-dioxide.

The structures and physical data of this compound and the compound obtained by the same are shown in table 2.

TABLE 1

Comp. Exp. R¹ R² n X¹ X² R³ Y⁴ m.p.(Recry.sol.^*3)

No.^*1 No.^*2 (℃)

01 2 H H 0 H H H S 241.0～242.5 (AcOEt)

02 2 n-Pr H 0 H H H S 172.0～174.0(AcOEt/Hex)

03 2 Me Me 0 H H H S 254.5～256.5(AcOEt)

04 2 Et Et 0 H H H S 242.0～243.5(AcOEt)

05 2 n-Pr n-Pr 0 H H H S 196.0～197.0(AcOEt)

06 2 n-Hex n-Hex 0 H H H S 138.5～140.0(AcOEt/Hex)

07^*42 n-Hex n-Hex 0 HHHHs 99.0-100.5 (Amplifier)^*5)

08^*42 n-Hex n-Hex 0 HHHHs 98.0-99.0 (Amplifier)^*5)

09 2 n-Hex n-Hex 0 8-F H H S 110.5～112.5(AcOEt/Hex)

10 2 n-Hex n-Hex 0 8-Cl H H S 123.0～124.5(AcOEt/Hex)

11 2 n-Hex n-Hex 0 8-Me H H S 139.5～141.5(AcOEt/Hex)

12 2 n-Hex h-Hex 0 8-F 10-F H S 148.0～149.0(AcOEt/Hex)

13 1 n-Hex CH₃(CH₂)₂NH(CH₂)₂ 0 H H H S 135.5～138.0(AcOEt)

14 1 n-Hex CH₃(CH₂)₂O(CH₂)₂ 0 H H H S 165.0～167.0(AcOEt)

15 2 CH₃O(CH₂)₂ CH₃O(CH₂)₂ 0 H H H S 206.0～208.0(AcOEt)

*1: compound numbering

*2: example numbers used for Compound Synthesis

*3: recrystallization solvents AcOEt ═ ethyl acetate, Hex ═ hexane, Et₂O ═ diethyl ether

*4: optically active form of compound 6 (compound 7 ═ compound 6, compound 8 ═ compound 6)

*5: the column was purified, dried, and left to crystallize.

In addition, R¹、R²In the formula, n-Hex ═ n-hexyl and n-Pr ═ n-propyl.

TABLE 2

Comp. Exp. R¹ R² n X¹ X² R³ Y⁴ m.p.(Recry.sol.^*3)

No.^*1 No.^*2 (℃)

16 3 n-Hex n-Hex 0 H H Me S 111.0～112.0(Hex)

17 3 n-Hex n-Hex 0 H H (CH₂)₂N(CH₂)₄ S 50.0～52.0(Hex)

18 3 n-Hex n-Hex 0 H H (CH₃)₂N(CH₃)₂ S 89.5～90.5(Et₂O/Hex)^*4

19 2 n-Hex n-Hex 1 H H H S 155.0～157.0(AcOEt/Hex)

20 5 n-Pr H 0 H H H SO₂ 296.5～298.0(AcOEt)

21 5 n-Pr n-Pr 0 H H H SO₂ 266.0～267.0(AcOEt/Hex)

22 5 n-Hex n-Hex 0 H H H SO₂ 156.0～157.0(AcOEt/Hex)

23 2 n-Hex n-Hex 0 H H H CH₂ 152.0～153.5(AcOEt/Hex)

27 2 n-Hex n-Hex 0 H H H OHCN 137.0～139.0(Et₂O/Hex)

28 2 n-Hex n-Hex 0 H H H AcN 145.0～146.0(AcOEt/Hex)

*1: compound numbering

*2: example numbers used for Compound Synthesis

*3: recrystallization solvents AcOEt ═ ethyl acetate, Hex ═ ethane, Et₂O ═ diethyl ether

*4: 1 hydrochloride salt

In addition, R¹、R²In the formula, n-Hex ═ n-hexyl and n-Pr ═ n-propyl.

TABLE 3

Comp Exp. R¹ R² n X¹ X² Y⁴ m.p.(Recry.sol.^*3)

No.^*1 No.^*2 (℃)

24 4 n-Et n-Et 0 H H CH₂206.5-207.5 (Amplifier)^*4)

25 4 n-Pr n-Pr 0 H H CH₂ 188.6～190.0(AcOEt/Hex)

26 4 n-Hex n-Hex 0 H H CH₂ 134.5～136.5(AcOEt)

*1: compound numbering

*2: example numbers used for Compound Synthesis

*3: recrystallization solvent AcOEt ═ ethyl acetate, Hex ═ hexane

*4: the column was purified, dried, and left to crystallize.

In addition, R¹、R²In the formula, n-Hex ═ n-hexyl and n-Pr ═ n-propyl.

Test example [ MDR receptor binding test ]

The pachytene fraction prepared from rat cerebral cortex was used as a receptor standard.

Use of³H]PK11195 as [ 2 ]³H]The ligand is identified.

Use of³H]The binding assay for labeled ligands was performed according to the following method described in Journal of pharmacological and Experimental Therapeutics, 262, 971 (1992).

Preparation of receptor standard: the cerebral cortex was homogenized with 10mM Hepes buffer (pH7.4) containing 0.32M sucrose in a volume 10 times its wet weight using a Teflon homogenizer. The homogenate was centrifuged at 900 Xg for 10 minutes. The resulting supernatant was centrifuged at 9000 Xg for 10 minutes. The sediment was suspended in Hepes buffer to a protein concentration of 1mg/ml, and centrifuged at 12000 Xg for 10 minutes. The obtained sediment was suspended in 50mM Hepes buffer (pH7.4) to obtain a crude mitochondrial fraction.

MDR binding assay: the mitochondrial standard (1.0mg protein/ml), (ii) is prepared³H]PK11195(2nM) and the test drug were reacted at 4 ℃ for 90 minutes.

After completion of the reaction, the reaction mixture was filtered by suction through a glass filter (GF/B) treated with 0.3% polyethyleneimine, and the reflection energy of the filter paper was measured by a liquid scintillation meter.

The combination of [ 10 ] MPK11195 and [ 2 ]³H]Non-specific binding of PK11195, the difference between total binding and non-specific binding was taken as specific binding. By allowing a certain concentration of [ alpha ], [³H]PK11195(2nM) reacts with the test drug at a varying concentration under the above-mentioned conditions to obtain an inhibition curve, and from the inhibition curve, 50% inhibition [ b ], [³H]PK11195 binding to the concentration of drug tested (IC)₅₀) The results are shown in Table 4.

TABLE 4

Comp.No.	MDRIC50(nM)
		020304050607080910111214151621222325262728	55.938.50.4430.3683.7642.32.1513.820.113.922.11.354.981.351.121.231.3573.942.335.112.6

Claims (1)

1. A nitrogen-containing tetracyclic compound represented by the formula or a pharmaceutically acceptable salt thereof.

In the formula, Y¹-Y²-Y³Represents N-C-N or formula C-NR³(in the formula, R³Represents a hydrogen atom, C_1-5Alkyl or C_2-10Nitrogen-containing alkyl group of (a); y is⁴Denotes S, SO₂、CH₂Or NR⁴(in the formula, R⁴Is represented by C_1-5Alkanoyl or C_1-5Alkyl) of (a); r¹And R²The same or different represent a hydrogen atom, C_1-10Alkyl of (C)_3-15Alkoxyalkyl or C_3-15Alkylaminoalkyl of (a); x¹And X²The same or different represent a hydrogen atom, C_1-5Alkyl of (C)_1-5N represents 0, 1 or 2.