RU2720510C2 - Coumarin, thiocoumarin and quinolinone derivatives, having anticonvulsant activity - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.SUBSTANCE: invention relates to derivatives of (coumarin-4-yl) aminocarboxylic acids, (thiocoumarin-4-yl) aminocarboxylic acids and (2-oxo-1,2-dihydroquinolin-4-yl) aminocarboxylic acids. In formulas 1.1–1.3, R is hydrogen or a nitro group; X is O, S or NH; Y is OH, OCH, OCH, NHor NHCH. Invention also relates to methods for preparing said compounds and a method of treating epilepsy and paroxysmal conditions using said compounds.EFFECT: can be used in treating epilepsy and paroxysmal conditions.13 cl, 5 tbl, 13 ex

Description

The invention relates to medicine, in particular to pharmacology and pharmacy, to the field of biologically active compounds, specifically to new groups of substituted (coumarin-4-yl) aminocarboxylic acids, (thiocoumarin-4-yl) aminocarboxylic acids, (2-oxo-1, 2-dihydroquinolin-4-yl) aminocarboxylic acids and their derivatives of the general formula (1.1-1.3):

where R may be a hydrogen atom or a nitro group; n may range from 3 to 6; X may be O (1.1), S (1.2), NH (1.3); Y may be O and NH; Z may be a hydrogen atom or a low molecular weight alkyl group,

which can be used to expand the arsenal of drugs used to treat epilepsy and paroxysmal activity that occurs in various pathologies. The invention also relates to methods for producing new groups of substituted (coumarin-4-yl) aminocarboxylic acids, (thiocoumarin-4-yl) aminocarboxylic acids, (2-oxo-1,2-dihydroquinolin-4-yl) aminocarboxylic acids and their derivatives.

State of the art

Epilepsy is a chronic disease characterized by repeated involuntary attacks of impaired motor, sensory, autonomic, mental, or mental functions resulting from excessive neural discharges in a group of cells in various regions of the brain. Attacks can range from a short loss of attention or muscle jerks to severe and prolonged cramps. Epileptic seizures depend on the developed epileptic system, including the formation of a dominant focus and the area of the involved brain structures in the pathological cycle. The classification of epileptic symptoms is diverse and includes a dichotomy both in duration of seizures and in form. Focal and generalized seizures are distinguished, which can include a wide variety of clinical signs. The treatment of epilepsy is to reduce the excitability of neurons in the epileptogenic focus, which can in principle be caused either by inhibition of activating neurons or activation of inhibitory nerve cells [Gennaro, A.R. (1995) Remington: The Science and Practice of Pharmacy, 19th Ed., Vol. II, Easton, PA, Mack Publishing Co., pp. 1164-1165]. Modern antiepileptic drugs include substances of various classes of chemical compounds and a mechanism of action, many of which have been known for a long time, but none of these drugs is universal, equally successfully used in any form of epileptic seizures. For example, carbamazepine is highly effective for seizures, but does not help with absences.

. Could X-Ray Analysis Explain for the Differing Antimicrobial and Antioxidant Activity of Two 2-Arylamino-3-Nitro-Coumarins? // J. Chem. Crystallogr. - 2011. - Vol. 41, №4. - P. 545-451]. Установлена способность N-замещенных 4-аминохинолин-2-онов связываться с NMDA рецептором и ингибировать нейротоксичность [Jung J. Synthesis and biological properties of 4-substituted quinolin-2(1H)-one analogues // J. Heterocycl. Chem. - 2003. - Vol. 40, №4. - P. 617-623]. Описаны некоторые 4,7-дизамещенные кумарины, которые являются ингибиторами моноаминоксидазы Б (МАО-Б) [L. Pisani. Fine molecular tuning at position 4 of 2H-chromen-2-one derivatives in the search of potent and selective monoamine oxidase В inhibitors // Eur. J. Med. Chem. - 2013, - Vol. 70. - P. 723-739]. Ряд производных 3-нитро-4-аминокумаринов обладает нейротропной активностью [Savel'ev V.L. Investigations of pyrans and related compounds. Synthesis of 4-amino-3-nitrocoumarins // Chem. Heterocycl. Compd. - 1973. - Vol. 9. - P. 44-47]. N-замещенные 4-амино-1-тиокумарины и 4-аминокумарины оказывают влияние на центральную нервную систему (ЦНС), потенцируя наркотический эффект тиопентала натрия, проявляют антагонизм с амфетамином на модели амфетаминовой гиперактивности на мышах (AHA) и противосудорожное действие в тесте антагонизма с коразолом [Pryanishnikova N.Т. Synthesis and study of the pharmacological properties of 3,4-diaminocoumarins and 3-substituted 4-aminothiocoumarins // Pharm. Chem. J. - 1978. - Vol. 12, №12. - P. 1590-1593]. Описаны производные кумарина обладающие обезболивающим, противовоспалительным и жаропонижающим действием [Пат. ВЕ-610413-А1. Amino-substituted coumarins and processes for their preparation // A. Geigy, заявл. 19.10.1961, опубл. 16.05.1962].Among the known derivatives of coumarin, thiocoumarin and quinoline, which are closest in structure to the claimed compounds, there are substances with different pharmacological activity. Coumarin derivatives are known to exhibit antimicrobial and fungicidal activity against microorganisms such as Staphylococcus aureus, Salmonella enterica, Sarcina Lutea, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Bacillus subtilis, Aspergillus niger Candida niger, Nigra Candida

. Could X-Ray Analysis Explain for the Differing Antimicrobial and Antioxidant Activity of Two 2-Arylamino-3-Nitro-Coumarins? // J. Chem. Crystallogr. - 2011 .-- Vol. 41, No. 4. - P. 545-451]. The ability of N-substituted 4-aminoquinolin-2-ones to bind to the NMDA receptor and inhibit neurotoxicity was established [Jung J. Synthesis and biological properties of 4-substituted quinolin-2 (1H) -one analog // J. Heterocycl. Chem. - 2003. - Vol. 40, No. 4. - P. 617-623]. Some 4.7-disubstituted coumarins are described which are monoamine oxidase B (MAO-B) inhibitors [L. Pisani. Fine molecular tuning at position 4 of 2H-chromen-2-one derivatives in the search of potent and selective monoamine oxidase B inhibitors // Eur. J. Med. Chem. - 2013, - Vol. 70. - P. 723-739]. A number of derivatives of 3-nitro-4-aminocoumarins have neurotropic activity [Savel'ev VL Investigations of pyrans and related compounds. Synthesis of 4-amino-3-nitrocoumarins // Chem. Heterocycl. Compd - 1973. - Vol. 9. - P. 44-47]. N-substituted 4-amino-1-thiocoumarins and 4-aminocoumarins affect the central nervous system (CNS), potentiating the narcotic effect of sodium thiopental, exhibit amphetamine antagonism in the mouse amphetamine hyperactivity model (AHA) and anticonvulsant effect in the c antagonism test Corazole [Pryanishnikova N.T. Synthesis and study of the pharmacological properties of 3,4-diaminocoumarins and 3-substituted 4-aminothiocoumarins // Pharm. Chem. J. - 1978. - Vol. 12, No. 12. - P. 1590-1593]. Coumarin derivatives having analgesic, anti-inflammatory and antipyretic effects are described [Pat. BE-610413-A1. Amino-substituted coumarins and processes for their preparation // A. Geigy, decl. 10/19/1961, publ. 05/16/1962].

The present invention is based on the task of creating compounds with high antiepileptic activity in the series of N-substituted 4-aminocoumarins, 4-amino-1-thiocoumarins and 4-aminoquinolin-2-ones. Based on this, a pharmacophore model is proposed that includes the following fragments: a coumarin, thiocoumarin or quinoline heterocycle with an electron-withdrawing substituent in 3 position and, possibly, in 5 position, and an ω-amino acid or its derivatives in 4 position. The goal can be achieved by obtaining compounds belonging to the groups of substituted (coumarin-4-yl) aminocarboxylic acids, (thiocoumarin-4-yl) aminocarboxylic acids, (2-oxo-1,2-dihydroquinolin-4-yl) aminocarboxylic acids and their derivatives of the general formula (1.1-1.3).

Compounds of the general formula (1.1-1.3), their properties and the preparation method are not described in the special and patent literature.

As examples of compounds of the formula (1.1-1.3) of the present invention are:

N- (3-nitrocoumarin-4-yl) -4-aminobutyric acid (1.1a),

N- (3-nitrocoumarin-4-yl) -6-aminohexanoic acid (1.1b),

N- (3,6-dinitrocoumarin-4-yl) -4-aminobutyric acid (1.1c),

N- (3-nitrocoumarin-4-yl) -4-aminobutyric acid methyl ester (1.1 g),

N- (3,6-dinitrocoumarin-4-yl) -4-aminobutyric acid methyl ester (1.1e),

N- (3-nitro-1-thiocoumarin-4-yl) -4-aminobutyric acid (1.2a),

N- (3-nitrothiocoumarin-4-yl) -4-aminobutyric acid methyl ester (1.2b),

N- (3-nitro-2-oxo-1,2-dihydroquinolin-4-yl) -4-aminobutyric acid methyl ester (1.3a)

N- (3-nitro-2-oxo-1,2-dihydroquinolin-4-yl) -4-aminobutyric acid (1.3b)

In accordance with the present invention, substituted (coumarin-4-yl) aminocarboxylic acids and their derivatives of the general formula (1.1) can be prepared according to the following general scheme.

3-Nitro-4-chlorocoumarins of the general formula (2), where R have the above meanings, are reacted with aminocarboxylic acids or their derivatives of the general formula (3), where n, Y and Z have the above meanings, in various solvents, such such as benzene, toluene, ethyl acetate, dioxane, ethanol, or the like. Reactions can be carried out both with an excess of compounds (3), and in the presence of bases, such as organic tertiary amines, for example, triethylamine, or pyridine.

, М., Trkovnik, М,

, А., 1984, Syntheses and testing of 4-chloro-3,6-dinitrocoumarin, a new derivatizing agent useful in thin layer chromatography of amines and amino acids, Acta Pharm. Jugosl., 34: 81].3-Nitro-4-chlorocoumarin (2a, R = H) is a commercially available reagent. 3,6-dinitro-4-chlorocoumarin (2b, R = NO ₂ ) is obtained from 4-hydroxycoumarin in accordance with the procedure described in the literature [

, M., Trkovnik, M,

, A., 1984, Syntheses and testing of 4-chloro-3,6-dinitrocoumarin, a new derivatizing agent useful in thin layer chromatography of amines and amino acids, Acta Pharm. Jugosl., 34: 81].

The structure of substances of the general formula (1.1) is confirmed by the data of ¹ H NMR spectra, and their purity - by the data of elemental analysis.

In accordance with the present invention, substituted (thiocoumarin-4-yl) aminocarboxylic acids and their derivatives of the general formula (1.2) can be prepared according to the following general scheme.

3-Nitro-4-chlorothiocoumarin (4) is reacted with aminocarboxylic acids or their derivatives of the general formula (3), where n, Y and Z have the above meanings, in various solvents such as benzene, toluene, ethyl acetate, dioxane ethanol, or the like. Reactions can be carried out both with an excess of compounds (3), and in the presence of bases, such as organic tertiary amines, for example, triethylamine, or pyridine.

3-Nitro-4-chlorothiocoumarin (4) is prepared according to the scheme:

The interaction of an excess of thiophenol (5) with malonic acid (6) when heated in a medium of phosphoryl chloride or thionyl chloride produces dithiophenyl ester of malonic acid (7). When compound 7 is heated in the presence of Lewis acids, such as, for example, aluminum halides and zinc halides, and in the presence of sodium chloride, 4-hydroxythiocoumarin is formed (8). Nitration of 4-hydroxythiocoumarin (8) by the action of nitric acid or a mixture of nitric acid with other acids, such as, for example, sulfuric and acetic acids, leads to the formation of 4-hydroxy-3-nitrothiocoumarin (9). Under the action of chlorinating agents, such as, for example, phosphoryl chloride or thionyl chloride, in polar aprotic solvents, compound (9) is converted to 3-nitro-4-chlorothiocoumarin (4). Suitable polar aprotic solvents are dimethylformamide or dimethyl sulfoxide.

The structure of substances of the general formula (1.2) is confirmed by the data of ¹ H NMR spectra, and their purity - by the data of elemental analysis.

In accordance with the present invention, substituted (2-oxo-1,2-dihydroquinolin-4-yl) aminocarboxylic acids and their derivatives of the general formula (1.3) can be prepared according to the following general scheme.

3-Nitro-4-chloroquinolin-2 (1H) -one (10) is reacted with aminocarboxylic acids or their derivatives of the general formula (3), where n, Y and Z have the above meanings, in various solvents, such as: benzene, toluene, ethyl acetate, dioxane, ethanol, or the like. Reactions can be carried out both with an excess of compounds (3), and in the presence of bases, such as organic tertiary amines, for example, triethylamine, or pyridine. The interaction of compounds 3 and 10 can also be carried out using pyridine as a solvent.

3-Nitro-4-chloroquinolin-2 (1H) -one (10) is obtained according to the scheme:

When an excess of aniline (11) interacts with malonic acid diethyl ether (12) when heated at a temperature of 100-230 ° С, malonic acid bisanilide is formed (13). When compound 13 is heated in the presence of Lewis acids, such as, for example, aluminum halides and zinc halides, or in the presence of polyphosphoric acid, 4-hydroxyquinolin-2 (1H) -one is formed (14). Nitration of 4-hydroxyquinolin-2 (1H) -one (14) by the action of nitric acid or a mixture of nitric acid with other acids, such as, for example, sulfuric and acetic acids, leads to the formation of 4-hydroxy-3-nitroquinolin-2 (1H) -one (15). Under the action of chlorinating agents, such as, for example, phosphoryl chloride or thionyl chloride, in the presence of an excess of quaternary ammonium salts in polar aprotic solvents, compound (15) turns into 3-nitro-4-chloroquinolin-2 (1H) -one (10). Suitable polar aprotic solvents are acetonitrile, dimethylformamide or dimethyl sulfoxide. Suitable quaternary ammonium salts are tetraalkylammonium halides, for example tetrabutylammonium bromide, trialkylbenzylammonium halides, for example tributylbenzylammonium bromide.

In accordance with the present invention, substituted (2-oxo-1,2-dihydroquinolin-4-yl) aminocarboxylic acids of the general formula (1.3 ″) can be prepared according to the following general scheme.

Alkyl esters of (2-oxo-1,2-dihydroquinolin-4-yl) aminocarboxylic acids of the general formula (1.3 '), where n has the above meanings, and Q may be OCH ₃ and OC ₂ H ₅ groups are reacted with alkali hydroxides or alkaline earth metals when heated in a mixture of water with alkyl alcohols, resulting in the formation of compounds 1.3 ''.

The structure of substances of the general formula (1.3) is confirmed by the data of ¹ H NMR spectra, and their purity - by the data of elemental analysis.

The experimental chemical part.

Example 1

N- (3-nitrocoumarin-4-yl) -4-aminobutyric acid (1.1a).

To a solution of 10 mmol (2.25 g) of 3-nitro-4-chlorocoumarin (2a) in 20 ml of dioxane was added 22 mmol (2.34 g) of 4-aminobutyric acid (3a). The reaction mass is boiled for two hours, and then cooled. The resulting precipitate is filtered off, washed with 10 ml of dioxane, three times with 10 ml of distilled water. The resulting material was recrystallized from 80 ml of ethanol to obtain 2.41 g of a yellow crystalline substance (yield 83%). Mp 210-211 ° C. ¹ H NMR spectrum (DMSO, δ, ppm, J / Hz): 1.90 (t, 2H, NHCH ₂ C H ₂ , ³ J ₁ = ³ J ₂ = 7.1); 2.30 (t, 2H, C H ₂ COOH, ³ J = 7.1); 3.18 (t, 2H, NHC H ₂ , ³ J = 7.1); 7.43 (m, 2H, NS (5), NS (6)); 7.73 (dd, 1H, NS (7), ³ J = 8.1); 8.33 (d, 1H, NS (8), ² J = 8.1); 8.43 (br s, 1H, NH). Found (%): C, 53.46; H, 4.24; N, 9.61. C ₁₃ H ₁₃ N ₂ O ₆ . Calculated (%): C, 53.63; H, 4.14; N, 9.59.

Example 2

N- (3-nitrocoumarin-4-yl) -6-aminohexanoic acid (1.1b).

Prepared in accordance with Example 1 from 3-nitro-4-chlorocoumarin (2a) and 6-aminohexanoic acid (3b). Yield 63% (yellow crystals). Mp 163-165 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 1.30 (m, 2H, C H ₂ CH ₂ CH ₂ COOH); 1.49 (m, 2H, C H ₂ CH ₂ COOH); 1.65 (m, 2H, NHCH ₂ C H ₂ ); 2.19 (t, 2H, C H ₂ COOH, ³ J = 7.1); 3.15 (m, 2H, NHC H ₂ ); 7.42 (m, 2H, NS (5), NS (6)); 7.73 (t, 1H, NS (7), ³ J = 7.6); 8.31 (d, 1H, HC (8), ² J = 7.6); 8.51 (br s, 1H, NH). Found (%): C, 56.21; H, 5.11; N, 8.72. C ₁₅ H ₁₅ N ₂ O ₆ . Calculated (%): C, 56.25; H, 5.04; N, 8.75.

Example 3

N- (3,6-dinitrocoumarin-4-yl) -4-aminobutyric acid (1.1c).

Prepared in accordance with Example 1 from 3,6-dinitro-4-chlorocoumarin (2b) and 4-aminobutyric acid (3a). Yield 74% (pale yellow crystals). Mp 183-185 ° C. ¹ H NMR spectrum (DMSO, δ, ppm, J / Hz): 1.91 (t, 2H, NHCH ₂ C H ₂ , ³ J ₁ = 7.2, ³ J ₂ = 6.8); 2.30 (t, 2H, CH ₂ CO, ³ J = 7.2); 3.17 (dt, 2H, NHC H ₂ , ³ J ₁ = 6.8, ³ J ₂ = 5.0); 7.62 (d, 1H, H (8), ³ J = 9.1); 8.50 (dd, 1H, H (7), ³ J = 9.1, ⁴ J = 2.3); 8.85 (br s, 1H, NH); 9.41 (d, 1H, H (5), ⁴ J = 2.3), 12.18 (br.s, 1H, OH). Found (%): C, 46.21; H, 3.50; N, 12.62. C ₁₃ H ₁₁ N ₃ O ₈ . Calculated (%): C, 46.30; H, 3.29; N, 12.46.

Example 4

N- (3-nitrocoumarin-4-yl) -4-aminobutyric acid methyl ester (1.1g).

To a solution of 10 mmol (2.25 g) of 3-nitro-4-chlorocoumarin (2a) in 80 ml of toluene, a thoroughly transferred two-phase mixture of 10 mmol (1.54 g) of 4-aminobutyric acid methyl ester hydrochloride (3c) in a minimum volume of water is added with stirring 8 ml) and 20 mol (2.02 g) of triethylamine. The mixture was stirred at room temperature for 30 minutes. The precipitate formed is filtered off, washed with 50 ml of 1% HCl and 100 ml of distilled water, dried on a filter to obtain 2.30 g of a pale yellow crystalline substance (yield 74%). Mp 125-126 ° C. ¹ H NMR spectrum (DMSO, δ, ppm, J / Hz): 1.92 (t, 2H, NHCH ₂ C H ₂ , ³ J ₁ = ³ J ₂ = 7.1); 2.38 (t, 2H, C H ₂ COOH, ³ J = 7.1); 3.19 (m, 2H, NHC H ₂ ); 3.55 (s, 3H, OCH ₃ ); 7.43 (m, 2H, NS (5), NS (6)); 7.74 (t, 1H, NS (7), ³ J = 8.3); 8.33 (d, 1H, NS (8), ² J = 8.3); 8.42 (br s, 1H, NH). Found (%): C, 54.78; H, 4.72; N, 9.06. C ₁₅ H ₁₅ N ₂ O ₆ . Calculated (%): C, 54.90; H, 4.61; N, 9.15.

Example 5

N- (3,6-dinitrocoumarin-4-yl) -4-aminobutyric acid methyl ester (1.1e).

Prepared in accordance with Example 4 from 3,6-dinitro-4-chlorocoumarin (2b) and 4-aminobutyric acid methyl ester hydrochloride (3c). Yield 79% (pale yellow crystals). Mp 130-131 ° C. ¹ H NMR spectrum (DMSO, δ, ppm, J / Hz): 1.94 (tt, 2H, NHCH ₂ C H ₂ , ³ J ₁ = 7.3, ³ J ₂ = 6.8); 2.40 (t, 2H, CH ₂ CO, ³ J = 7.3); 3.17 (dt, 2H, NHC H ₂ , ³ J ₁ = 6.8, ³ J ₂ = 5.0); 3.57 (s, 3H, OCH ₃ ); 7.64 (d, 1H, H (8), ³ J = 9.1); 8.51 (dd, 1H, H (7), ³ J = 9.1, ⁴ J = 2.5); 8.83 (t, 1H, NH, ³ J = 5.0); 9.4 (d, 1H, H (5), ⁴ J = 2.5). Found (%): C, 48.08; H, 3.62; N, 12.11. C ₁₄ H ₁₃ N ₃ O ₈ . Calculated (%): C, 47.87; H, 3.73; N, 11.96.

Example 6

N- (3-nitro-1-thiocoumarin-4-yl) -4-aminobutyric acid (1.2a).

6.1. Malic acid dithiophenyl ester (7).

75 mmol (7.81 g) of malonic acid, 80 mmol (12.27 g) of phosphoryl chloride and 150 mol (16.52 g) of thiophenol are mixed. The resulting solution is boiled in an oil bath for 40 minutes. Cool the solution and add 80 ml of ethanol. The precipitate formed is filtered off and washed with 50 ml of ethanol to obtain pale blue crystals. After recrystallization from 30 ml of ethanol, 17.90 g of a white crystalline substance are obtained (82% yield). T. pl. 94-95 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm): 4.33 (s, 2H, CH ₂ ); 7.36-7.54 (m, 10H, 2Ph).

6.2. 4-Hydroxy-1-thiocoumarin (8).

To 40 mmol (11.52 g) of malonic acid dithiophenyl ether (7), 120 mmol (16.00 g) of aluminum chloride and 80 mmol (4.68 g) of sodium chloride are added and mixed. The mixture is heated to 195 ° C and maintain the temperature for 25 minutes. After cooling, 200 ml of 1% hydrochloric acid and 200 ml of ethyl acetate are poured into the reaction mass, the mixture is stirred. The organic layer was separated, 200 ml of 10% sodium hydroxide solution was added to it, the mixture was stirred. The aqueous layer is separated and acidified to pH = 4, the precipitate is filtered off, washed twice with 40 ml of distilled water. Obtain 3.56 g of a gray-orange crystalline substance (yield 62%). Mp 210-211 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 3.61 (br s, 1H, OH). 6.14 (s, 1H, HC (3)); 7.48 (m, 2H, NS (5), NS (6)); 7.59 (t, 1H, NS (7), ³ J = 8.0); 8.15 (d, 1H, NS (8), ² J = 8.0).

6.3. 4-Hydroxy-3-nitro-1-thiocoumarin (9).

To a suspension of 18 mmol (3.21 g) of 4-hydroxythiocoumarin (8) in 80 ml of glacial acetic acid was added a mixture of 20 mmol (1.40 ml) of 65% nitric acid and 20 mmol (1.14 ml) of 96% sulfuric acid. The mixture was stirred at room temperature for 3 hours. The precipitate formed is filtered off, washed twice with 10 ml of distilled water, and dissolved in boiling toluene. The hot solution was passed through activated carbon and evaporated to dryness to obtain 3.78 g of a yellow powder (yield 92%). Mp 149-152 ° C. ¹ H NMR spectrum (DMSO, δ, ppm, J / Hz): 7.38 (m, 2H, NS (5), NS (6)); 7.44 (t, 1H, NS (7), ³ J = 8.1); 8.17 (d, 1H, NS (8), ³ J = 8.1).

6.4. 3-Nitro-4-chloro-1-thiocoumarin (4).

4 mmol (0.373 ml) of phosphoryl chloride are dissolved in 2.5 ml of dimethylformamide and stirred for 30 minutes. 4 mmol (0.89 g) of 4-hydroxy-3-nitrothiocoumarin (9) in 9.6 ml of dimethylformamide was added to the solution, and the resulting mixture was stirred for 2 hours. 40 ml of cold water are added to the reaction mixture, the precipitate formed is filtered off, washed three times with 10 ml of distilled water and recrystallized from 10 ml of ethanol. Obtain 0.85 g of dark yellow crystals (yield 88%). Mp 171-173 ° C. ¹ H NMR spectrum (DMSO, δ, ppm, J / Hz): 7.77 (m, 1H, HC (5)); 7.89 (m, 1H, HC (6)); 7.94 (m, 1H, NS (7)), 8.38 (d, 1H, NS (8), ² J = 8.1).

6.5. N- (3-nitro-1-thiocoumarin-4-yl) -4-aminobutyric acid (1.2a).

Prepared in accordance with Example 1 from 3-nitro-4-chloro-1-thiocoumarin (4) and 4-aminobutyric acid (3a). Yield 59% (yellow crystals). Mp 215-216 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 1.90 (m, 2H, NHCH ₂ C H ₂ ); 2.30 (t, 2H, CH ₂ COOH, ³ J = 7.1); 3.18 (m, 2H, NHC H ₂ ); 7.43 (m, 2H, NS (5), NS (6)); 7.73 (t, 1H, NS (7), ³ J = 8.1); 8.33 (d, 1H, NS (8), ³ J = 8.1); 8.43 (br s, 1H, NH). Found (%): C, 50.64; H, 3.69; N, 9.33; S, 10.41. C ₁₃ H ₁₁ N ₂ O ₅ S. Calculated (%): C, 50.65; H, 3.92; N, 9.09; S, 10.40.

Example 7

N- (3-nitrothiocoumarin-4-yl) -4-aminobutyric acid methyl ester (1.2b).

Prepared in accordance with Example 4 from 3-nitro-4-chloro-1-thiocoumarin (4) and 4-aminobutyric acid methyl ester hydrochloride (3c). Yield 77% (pale yellow crystals). Mp 125-126 ° C. ¹ H NMR spectrum (DMSO, δ, ppm, J / Hz): 1.92 (t, 2H, NHCH ₂ C H ₂ , ³ J ₁ = ³ J ₂ = 7.1); 2.38 (t, 2H, C H ₂ COOH, ³ J = 7.1); 3.19 (m, 2H, NHC H ₂ ); 3.55 (s, 3H, OCH ₃ ); 7.43 (m, 2H, NS (5), NS (6)); 7.74 (t, 1H, NS (7), ³ J = 8.3); 8.33 (d, 1H, NS (8), ³ J = 8.3); 8.42 (br s, 1H, NH). Found (%): C, 52.06; H, 4.22; N, 8.87; S, 9.87. C ₁₄ H ₁₄ N ₂ O ₅ S. Calculated (%): C, 52.17; H, 4.38; N, 8.69; S, 9.95.

Example 8

N- (3-nitro-2-oxo-1,2-dihydroquinolin-4-yl) -4-aminobutyric acid methyl ester (1.3a).

8.1. Malonic acid bis anilide (13).

A mixture of 200 mmol (18.64 g) of aniline (11) and 100 mmol (16.02 g) of malonic acid diethyl ester (12) is heated at a temperature of 200 ° C for 8 hours. After cooling the reaction mixture, the precipitate formed is filtered off and recrystallized from ethanol to obtain 19.3 g of a white powder (76% yield). Mp 228-230 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 3.48 (s, 2H, CH ₂ ); 7.07 (dd, 2H, 2 H (4), ³ J ₁ = 7.6, ³ J ₂ = 7.4); 7.32 (dd, 4H, 2 N (3), 2 N (5), ³ J ₁ = 7.6, ³ J ₂ = 8.1); 7.60 (d, 4H, 2H (2), 2H (6), ³ J = 8.1), 10.17 (s, 2H, 2NH).

8.2. 4-Hydroxyquinolin-2 (1H) -one (14).

A mixture of 50 mmol (12.72 g) of malonic acid bisanilide (13) and 80 g of polyphosphoric acid is heated at 215 ° C for 5 hours and poured into 400 g of ice. The precipitate formed is filtered off, washed with water until neutral and dried, yielding 5.88 g of a pale yellow powder (yield 73%). Mp > 300 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 5.74 (s, 1H, HC (3)); 7.13 (dd, 1H, H (6), ³ J ₁ = 8.0, ³ J ₂ = 7.1); 7.27 (d, 1H, H (5), ³ J = 8.0); 7.48 (dd, 1H, H (7), ³ J ₁ = 8.0, ³ J ₂ = 7.1); 7.77 (d, 1H, H (8), ³ J = 8.0); 11.96 (s, 1H, NH).

8.3. 4-Hydroxy-3-nitroquinolin-2 (1H) -one (15).

A suspension of 20 mmol (3.22 g) of 4-hydroxyquinolin-2 (1H) -one (14) in 30 ml of nitric acid (d = 1.33 g / ml) was stirred at room temperature for 10 minutes and at a temperature of 75 ° C for 20 minutes. The resulting solution was poured into 150 ml of ice water. The precipitate formed is filtered off, washed with water until neutral and dried, yielding 3.83 g of an orange powder (93% yield). Mp 243-245 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 7.27 (m, 2H, H (5), H (6)); 7.65 (m, 1H, H (7)); 8.02 (d, 1H, H (8), ³ J = 7.6); 11.96 (s, 1H, NH).

8.4. 3-Nitro-4-chloroquinolin-2 (1H) -one (10).

To a solution of 60 mmol (21.38 g) of tri-n-butylbenzylammonium bromide in 60 ml of acetonitrile, 15 mmol (3.09 g) of 4-hydroxy-3-nitroquinolin-2 (1H) -one (15) and 36 mmol (5.52 g) of phosphoryl chloride are added . The mixture is stirred at a temperature of 40 ° C for 30 minutes, after which it is refluxed for 22 minutes and evaporated to dryness. 60 ml of water are added to the residue and the mixture is stirred at room temperature for 3 hours. The precipitate formed is filtered off, washed with water until neutral and recrystallized from 20 ml of acetone to obtain 2.02 g of a yellow powder (60% yield). Mp 231-233 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 7.46 (m, 2H, H (5), H (6)); 7.80 (m, 1H, H (7)); 8.01 (d, 1H, H (8), ³ J = 7.9); 11.03 (s, 1H, NH).

8.5. N- (3-nitro-2-oxo-1,2-dihydroquinolin-4-yl) -4-aminobutyric acid methyl ester (1.3a).

To a solution of 1.25 mmol (0.28 g) of 3-nitro-4-chloroquinolin-2 (1H) -one (10) in 10 ml of pyridine, 5 mmol (0.77 g) of 4-aminobutyric acid methyl ester hydrochloride was added (3c). The reaction mass is refluxed for 3 hours. The solvent was evaporated to dryness, 15 ml of water was added to the residue, and the product was extracted twice with 15 ml of dichloromethane. The organic layer was evaporated to dryness and the residue was recrystallized from ethanol to obtain 0.20 g of a dark yellow powder (63% yield). Mp 208-210 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 1.98 (m, 2H, NHCH ₂ C H ₂ ); 2.36 (t, 2H, C H ₂ COOH, ³ J = 7.1); 3.11 (m, 2H, NHC H ₂ ); 3.55 (s, 3H, OCH ₃ ); 7.26 (m, 2H, NS (5), NS (6)); 7.42 (s, 1H, NH (Ar)); (7.58 (m, 1H, HC (7)); 8.21 (d, 1H, HC (8), ³ J = 8.1); 11.53 (br s, 1H, NH).

Example 9

N- (3-nitro-2-oxo-1,2-dihydroquinolin-4-yl) -4-aminobutyric acid (1.3b).

A mixture of 0.66 mmol (0.2 g) of N- (3-nitro-2-oxo-1,2-dihydroquinolin-4-yl) -4-aminobutyric acid methyl ester (1.3a), 1.3 mmol (52 mg) of sodium hydroxide, 20 ml of water and 20 ml of methanol are refluxed for 40 minutes, cooled to room temperature, acidified with 1% hydrochloric acid until acidic, the precipitate formed is filtered off, washed with water and dried, yielding 0.12 g of a dark yellow powder (yield 64 %). Mp 119-120 ° C. ¹ H NMR Spectrum (DMSO, δ, ppm, J / Hz): 1.85 (m, 2H, NHCH ₂ C H ₂ ); 2.28 (t, 2H, CH ₂ COOCH ₃ , ³ J = 7.2); 3.08 (m, 2H, NHC H ₂ ); 7.24 (m, 1H, HC (6)); 7.27 (d, 1H, H (8), ³ J = 7.6); 7.43 (m, 1H, N H CH ₂ ); 7.58 (m, 1H, HC (7)); 8.21 (d, 1H, HC (5), ³ J = 8.2); 11.53 (br s, 1H, NH (quinoline)), 12.16 (br s, 1H, COOH). Found (%): C, 53.58; H, 4.45; N, 14.40. C ₁₃ H ₁₃ N ₃ O ₅ . Calculated (%): C, 53.61; H, 4.50; N, 14.43.

The experimental pharmacological part.

The anticonvulsant activity of the new compounds was studied on conventional models of primary generalized seizures in experiments on mice and rats.

Experimental animals were obtained from the Stolbovaya nursery, GU NTsBMT (Moscow region). The keeping of animals corresponded to the rules of laboratory practice (GLP) and normative documents “Sanitary rules for the construction, equipment and maintenance of vivariums” approved by the Chief State Sanitary Doctor on 04/06/1973 No. 1045-73 and the Order of the Ministry of Health and Social Development of the Russian Federation of August 23, 2010 No. 708n “On approval of the Laboratory Practice Rules”.

Statistical processing of the results was performed using MS Excel 2010 and BioStat 2009 (Analyst Soft Inc.). The normality of the data distribution was determined by the Shapiro-Wilk criterion. The significance of differences between the groups was determined using nonparametric criteria: Kruskal-Wallis and Fisher's exact test.

Example 1

Anticonvulsant effects of coumarin, thiocoumarin and quinolinone derivatives on the model of primary generalized seizures caused by maximum electroshock.

The experiments were performed on white outbred male mice weighing 20-26 g. Each dose of the compound was studied in 8-10 animals. The method of maximum electroshock (MES) simulates primary generalized convulsions - the so-called “large” (Grantmal) convulsive seizures and is a basic test in assessing the effect of substances with anticonvulsant activity (Voronina T.A., Nerobkova L.N. Methodological guidelines for the study of anticonvulsant activity of pharmacological substances. "Guidelines for preclinical studies of drugs" Part. 1. FSBI "NTsEMSP". Moscow, GrifiK Publishing House, 2012, Chapter 14, 235-250; Loscher et al., The role of technical, biological and pharmacological factors in the labor atory evaluation of anticonvulsant drugs. II. Maximal electroshock seizure models, Epilepsy Res., 1991, v. 8, p. 79-94; Swinyard EA-Laboratory evaluation of antiepileptic drugs. Reviewof laboratory methods, Epilepsia, 1969, v. 10, pp. 107-119).

The maximum electroconvulsive seizure (MES) was created using a certified Rodent Shocker RS, type 221 (Harvard Apparatus, GmbH, Germany). Animals received electrical stimuli through special corneal electrodes (500/300 V / mA mode: 144 mA, duration 0.3 s). The following indicators were recorded: tonic extension of the hind and forelimbs and animal death. The anticonvulsant effect of the claimed compounds was evaluated by their ability to prevent the development of tonic extension and the death of animals. Compounds were administered intraperitoneally 40 minutes before MES.

It was found that conducting MES caused tonic extension and the death of 75-90% of mice. Compound 1.1a in the dose range of 20–40 mg / kg did not significantly affect seizures and the survival rate in the MES antagonism test, and in doses from 60 to 80 mg / kg it reduced the number of animals with tonic extension and increased the number of surviving animals compared with control (table 1).

Compound 1.1 g in doses from 1 to 60 mg / kg did not have a pronounced effect on mouse cramps caused by MES. Only at doses of 10 and 30 mg / kg under the influence of compound 1.1 g was an increase in the survival rate compared with the control up to 38% (table 1). Compound 1,2a changed the survival curve as a function of dose, and these changes were domed. Thus, at doses of 20 and 40 mg / kg, compound 1.2a helped to eliminate tonic extention and increased the survival of mice to 50 and 63%, respectively, while in the dose range from 60 to 120 mg / kg, the effectiveness of compound 1.2a decreased in terms of survival to 38% (table 1). Compounds 1.1c, 1.1d in doses of 10 to 40 mg / kg and compound 1.2b in doses of 10 to 60 mg / kg did not significantly affect seizures and survival in the MES antagonism test. Compound 1.3a at a dose of 12.5 mg / kg increased the survival rate relative to the control to 63%, but with an increase in dose to 25 and 50 mg / kg, its anticonvulsant effect was not recorded (Table 1).

Note: * - significance of differences from the control group, at p≤0.05 (Fisher's exact test).

Thus, it was found that in the test of antagonism with MES, compound 1.1a in the dose range from 60 to 80 mg / kg increases the number of surviving animals up to 60% compared with the control group, the survival rate of which was 10%. Compounds 1.2a and 1.3a showed activity in the antagonism test with MES at doses of 40 mg / kg and 12.5 mg / kg, respectively, at the level of the trend.

Example 2

Anticonvulsant effects of coumarin, thiocoumarin and quinolinone derivatives on the model of primary generalized seizures caused by corazole.

The experiments were performed on white outbred male mice weighing 20-26 g. Each dose of the compound was studied in 8-10 animals. The antagonism test with corazole (pentylenetetrazole, Sigma-Aldrich, USA), a GABA _A receptor antagonist, is the basic method for evaluating the effects of substances with anticonvulsant activity (Voronina TA, Nerobkova LN Methodological guidelines for the study of the anticonvulsant activity of pharmacological substances. " Guidelines for preclinical studies of drugs "Part. 1. FSBI" NTsEMSP ". Moscow, GrifiK Publishing House, 2012, Chapter 14, 235-250; Loscher et al., The role of technical, biological and pharmacological factors in the laboratory evaluation of anticonvulsant drugs. III. Pentylenetetrazolseizuremodels. Epilepsy Res., 1991, v. 8, p. 171-189). In this technique, convulsions are caused by chemical attack and simulate primary-generalized convulsions in the so-called “small” (Petit mal) seizures. Test groups of animals were injected intraperitoneally with the test substances dissolved in physiological saline 40 minutes before corazole. Control animals were injected intraperitoneally with physiological saline in an equivalent volume. To obtain a convulsive seizure, animals were injected subcutaneously in the cervical spine with a dose of 100 mg / kg, causing seizures in 97% of mice. Animals were observed for 30-60 minutes. after injection of corazole. The number of dead animals was recorded.

It was found that in control animals after administration of corazole at a dose of 100 mg / kg, convulsive manifestations developed in the following sequence. 1. One or more myoclonic twitches of the whole body - 100% of mice. 2. Repeated clonic cramps of the front and / or hind limbs lasting more than 3 seconds without loss of the overturning reflex - 100% of mice. 3. Generalized clonic convulsions of the front and hind limbs with the loss of the overturning reflex - 90% of mice. 4. The death of animals - 100% of mice.

It was found that compounds 1.1a in the dose range from 20 to 80 mg / kg and 1.1c in the dose range from 10 to 40 mg / kg did not prevent the development of convulsions caused by corazole and the death of animals (Table 2). Compound 1.1 g at a dose of 40 mg / kg helped to eliminate convulsive manifestations caused by corazole and death in 38% of mice (table 2). Compound 1.1d exerted the most pronounced protective effect in all studied doses from 10 to 40 mg / kg, preventing death in 50-63% of animals. Compounds 1.2a and 1.2b also showed anticonvulsant effects in doses of 10 to 40 mg / kg, increasing the survival of mice to a maximum of 50%. Compound 1.3a showed anticonvulsant activity at a dose of 12.5 mg / kg, increasing the survival of mice to 56% (Table 2).

Thus, it can be noted that the most pronounced anticonvulsant effect in this experiment is possessed by compound 1.1d at doses of 10, 20 and 40 mg / kg, compounds 1.2a, 1.2b at a dose of 10 mg / kg and 1.3a at a dose of 12.5 mg / kg, which not only increase the latent period of the onset of the first convulsive attack, but also prevent the development of seizures caused by corazole and the death of animals.

Note: * - significance of differences from the control group, at p≤0.05 (Fisher's exact test); # - a tendency to the reliability of differences from the control group, at p≤0.1 (Fisher's exact test).

Thus, the studies showed that among the studied compounds, compound 1.1d in the doses of 20 and 40 mg / kg has the highest anticonvulsant activity, preventing death in 63% of animals. Compounds 1.2a, 1.2b at a dose of 10 mg / kg and 1.3a at a dose of 12.5 mg / kg statistically reliably prevent the development of generalized convulsions caused by corazole and the death of 50-56% of animals.

Example 3

Anticonvulsant effect of N- (3-nitrocoumarin-4-yl) -4-aminobutyric acid (1.1a) on primary-generalized seizures caused by bemegrid in rats with chronically implanted electrodes in various brain structures.

Primarily generalized epileptiform activity (EPA) in rats with chronically implanted electrodes was induced by intramuscular injection of a 0.5% solution of bemegride at a dose of 10 mg / kg. The operation of implantation of chronic electrodes was performed under nembutal anesthesia (45 mg / kg) 5 days before the experiment. The electrodes were implanted in the sensorimotor cortex, the dorsal hippocampus (field CA3), the caudate nucleus, and the lateral hypothalamus. Brain biopotentials were recorded on a 21-channel Neurosensor neurograph operating on the basis of IBM-PC 586 with installed filters at 32 Hz, with a time constant (0.03 s) and with recording digital computer EEG for subsequent processing of the electrogram data of the structures under study. Compound 1.1a was administered 30 minutes before the administration of bemegrid, so that its peak was at the peak of bemegrid activity.

Bemegrid at a dose of 10 mg / kg caused a distinct EPA in the electrograms of all the studied structures (sensorimotor cortex, dorsal hippocampus, caudate nucleus and lateral hypothalamus). Epileptipharmic activity appeared in all animals as early as 5 minutes after the administration of epileptogen and after 15 minutes reached maximum values.

Preliminary administration of compound 1.1a at a dose of 50 mg / kg caused significant inhibition of EPA caused by bemegrid in the sensorimotor cortex, caudate nucleus, dorsal hippocampus, and hypothalamus. A significant decrease in the number of EPA discharges in the sensorimotor cortex and caudate nucleus was observed already 5 minutes after the introduction of bemegrid, reaching a maximum effect after 30 minutes, which was expressed in a reduction (2-2.7 times) in the number of EPA discharges in all studied structures (Table 3 )

* - difference from background indicators, at p≤0.05 ** at p≤0.01 (Kruskal-Wallis test)

Thus, the preliminary administration of compound 1.1a at a dose of 50 mg / kg reduces the severity of epileptipharmic activity caused by the introduction of bemegrid, with a maximum effect in the hippocampus, which was observed by reducing the number of discharges by 2.7 times.

Example 4

The anticonvulsant effect of N- (3-nitrocoumarin-4-yl) -4-aminobutyric acid (1.1a) on partial (focal) secondary-generalized convulsions in a chronic experiment in rats with a chronic cobalt-induced epileptogenic focus.

The studies were performed on male outbred adult white rats weighing 220-250 g. The study was conducted using the technique of creating a chronic epileptogenic focus caused by the application of cobalt, which models partial (focal) and secondary-generalized seizures in a chronic experiment. The technique is widely used to study the mechanisms of action of anticonvulsants in Russia and abroad (Avakyan G.N., Nerobkova L.N., Voronina T.A., Markina N.V. Mitrofanov A.A. Effect of carbamazepine on structural-functional bonds in the development of the epileptic system // Experimental and Clinical Pharmacology, 2002, No. 2, pp. 7-10; Bregman, F. Le Saux, S. Trottier 1, P. Chauvel 1, and Y. Maurin. Chronic Cobalt-induced Epilepsy: Noradrenaline Ionophoresis and Adrenoceptor Binding Studies in the Rat Cerebral Cortex. J. Neural Transmission, 1985, v. 63, p. 109-118) and is recommended by the Guidelines for Preclinical Drug Research funds, FBSI "NTsESMP" of the Ministry of Health and Social Development of Russia "(Voronina TA, Nerobkova LN Methodological guidelines for the study of anticonvulsant activity of pharmacological substances." Guidelines for preclinical studies of drugs ", M., edition of FBSU" NTsESMP "Ministry of Health and Social Development Russia, 2012, part 1, chapter 14, pp. 235-250).

The operations of implanting long-term electrodes into rat brain structures (into the motor zone of the left and right hemisphere cortex, the dorsal hippocampus, the lateral nuclei of the hypothalamus, and the caudate nucleus) were performed using a stereotactic device along the coordinates of the rat brain atlas (Bures et al .. 1960). Electrode implantation operations were performed under chloral hydrated anesthesia (300 mg / kg). The indifferent electrode used in monopolar recording was placed in the nasal bone of the skull. The bioelectric activity was recorded under conditions of free movement of the animal in the experimental chamber. In order to avoid artifacts from the movement of the pins, special spring contacts were used. Brain biopotentials were recorded on a 21-channel Neurosensor neurograph operating on the basis of IBM-PC 586 with installed filters at 32 Hz, with a time constant (0.03 s) and recording digital computer EEG for subsequent data processing.

The epileptogenic focus was created by the application of metallic cobalt powder on the surface of the motor region of the cortex of the left brain of rats. For this purpose, a trepanation hole was drilled into the bones of the skull, into which a glass cannula with cobalt powder was inserted (the diameter of the cannula corresponded to the diameter of the hole and did not exceed 1 mm). The cannula descended to the surface of the cortex (the dura mater was previously opened with a thin injection needle). Application of cobalt on the cerebral cortex of the brain causes hyperactivity of neurons localized at the injection site, which is expressed in the appearance of epileptiform paroxysmal discharges on the EEG. The emerging epileptogenic focus is the beginning of the development of a dynamic, constantly becoming more complex structural and functional system. The functional organization of this system is characterized by the presence of a determinant and dependent foci; the determinant focus enhances and synchronizes the activity of other foci, combining them into a single complex. Several stages are distinguished in the development of the epileptic system caused by the application of cobalt on the sensorimotor cortex of the brain. The main ones are the stage of formation of primary and secondary epileptogenic foci 24-48 hours after surgery and the stage of generalized epileptiform activity (EPA) in various brain structures with a stable level of synchronized paroxysmal discharges 5-6 days after application of cobalt (second stage of development of EPA) .

In all rats, the epileptogenic focus was localized in the sensorimotor region of the cortex of the left hemisphere in the following coordinates: 1 mm forward from bregma and 1 mm to the side of the sagittal suture. The dynamics of EPA in rats with a cobalt epileptogenic focus was studied for 7-8 days after the application of cobalt to the sensorimotor cortical zone. EEG registration was started 48 hours after the application of cobalt (the first stage of the development of the epileptic system and was performed on the 5-6th day (stage of generalization of EPA). The effect of compound 1.1a at a dose of 50 mg / kg on the epileptiform activity of the brain of rats was studied with a single intraperitoneal administration in various stages of the formation of the epileptic system after background recording.After administration of the substance, the EEG was recorded for 120 minutes.

Registration of the background EEG of animals 48 hours after the operation (stage 1) revealed the formation of foci of epileptic activity (EpA) in all studied structures with the largest number and duration of discharges in the ipsilateral cortex, caudate nucleus, and contralateral hippocampus (Table 4).

A study of the effect of compound 1.1a at a dose of 50 mg / kg on the epileptiform activity of rat brain showed that at the first stage of the formation of the epileptic system, 1 hour after a single injection, the number of EPA discharges (on average per minute) decreased statistically significantly in electrocorticograms of ipsi and the contralateral cortex, caudate nucleus, hypothalamus and contralateral hippocampus. Moreover, the greatest severity of the effect (according to the number of discharges) was determined in the ipsi and contralateral cortex, and a shortening of the average duration of discharges in these structures was also observed (Table 4).

* - difference from background indicators, at p≤0.05 (Kruskal-Wallis test)

At the 2nd stage of development of EPA (5-6 days), with persistently formed generalized EPA, a single injection of compound 1.1a at a dose of 50 mg / kg caused a significant decrease in the number and duration of discharges in electrograms of the ipsilateral cortex, but did not change these indicators relative to the background values in the contralateral cortex, lateral hypothalamus, ipsi, contralateral hippocampus and caudate nucleus. (Table 5).

Thus, under the conditions of the partial (focal) epilepsy technique simulating secondary generalized convulsions in a chronic experiment in rats with a chronic cobalt-induced epileptogenic focus, the coumarin derivative - compound 1.1a at a dose of 50 mg / kg has a pronounced anticonvulsant effect on primary epileptic foci in the ipsi, contralateral cortex, hypothalamus, contralateral hippocampus and caudate nucleus, statistically significantly reducing both the number of convulsive discharges and their duration at the 1st stage of ra power ES. Compound 1.1a does not affect the secondary epileptic foci that developed in the 2nd stage of EPA generalization in such structures as the hypothalamus, ipsi and contralateral hippocampus, caudate nucleus, and contralateral cortex. The statistically significant effect of compound 1.1a is detected only in the ipsilateral cortex, which is recorded by a decrease in the number and duration of discharges in this structure.

* - difference from background indicators, at p≤0.05 (Kruskal-Wallis test)

Thus, it was found that the derivatives of coumarin, thiocoumarin and quinolinone have a wide range of anticonvulsant effects, eliminating primary generalized convulsions in antagonism tests with maximum electroshock (MES), corazole. The anticonvulsant effect of the claimed compounds is observed in a wide range of doses (from 10 to 80 mg / kg). A coumarin derivative, compound 1.1a, at a dose of 50 mg / kg reduces the severity of epileptipharmic activity caused by the introduction of bemegrid, with a maximum effect in the hippocampus. Under the conditions of the partial (focal) epilepsy technique that simulates second-generalized convulsions in a chronic experiment in rats with a chronic cobalt-induced epileptogenic focus, the coumarin derivative - compound 1.1a at a dose of 50 mg / kg has a pronounced anticonvulsant effect on the epileptic foci in ipsi, contralateral cortex, hypothalamus, contralateral hippocampus and caudate nucleus in the first stage of ES development, whereas in the second stage of EPA generalization, a statistically significant effect is detected in ipsil teral cortex.

Claims (26)

1. Substituted (coumarin-4-yl) aminocarboxylic acids, (thiocoumarin-4-yl) aminocarboxylic acids, (2-oxo-1,2-dihydroquinolin-4-yl) aminocarboxylic acids and their derivatives of the general formula (1.1-1.3)

, where R may be a hydrogen atom or a nitro group; n may range from 3 to 6; X may be O (1.1), S (1.2), NH (1.3); Y may be OH, OCH ₃ , OS ₂ H ₅ , NH ₂ , NHCH ₃ groups. 2. The compound according to claim 1, which is N- (3-nitrocoumarin-4-yl) -4-aminobutyric acid (1.1a). 3. The compound according to claim 1, which is N- (3,6-dinitrocoumarin-4-yl) -4-aminobutyric acid (1.1c). 4. The compound according to claim 1, which is a methyl ester of N- (3-nitrocoumarin-4-yl) -4-aminobutyric acid (1.1 g). 5. The compound according to claim 1, which is a methyl ester of N- (3,6-dinitrocoumarin-4-yl) -4-aminobutyric acid (1.1e). 6. The compound according to claim 1, which is N- (3-nitro-1-thiocoumarin-4-yl) -4-aminobutyric acid (1.2a). 7. The compound according to claim 1, which is a methyl ester of N- (3-nitrothiocoumarin-4-yl) -4-aminobutyric acid (1.2b). 8. The compound according to claim 1, which is a methyl ester of N- (3-nitro-2-oxo-1,2-dihydroquinolin-4-yl) -4-aminobutyric acid (1.3a). 9. The compound according to claim 1, which is N- (3-nitro-2-oxo-1,2-dihydroquinolin-4-yl) -4-aminobutyric acid (1.3b). 10. The method of obtaining (coumarin-4-yl) aminocarboxylic acids and their derivatives of the General formula (1.1), where R, n and Y have the above values according to claim 1, namely that 3-nitro-4-chlorocoumarins of the general formula (2)

, where R have the above meanings, enter into interaction with aminocarboxylic acids or their derivatives of General formula (3)

, where n and Y have the above meanings in various solvents, such as benzene, toluene, ethyl acetate, dioxane, ethanol, or the like, while the reactions can be carried out both with an excess of compounds (3) and in the presence of bases, such as organic tertiary amines for example triethylamine, or pyridine. 11. The method of obtaining (thiocoumarin-4-yl) aminocarboxylic acids and their derivatives of the general formula (1.2), where R is a hydrogen atom, n and Y have the above values according to claim 1, namely, that 3-nitro-4- chlorothiocoumarin (4)

enter into interaction with aminocarboxylic acids or their derivatives of General formula (3)

, where n and Y have the above meanings in various solvents, such as benzene, toluene, ethyl acetate, dioxane, ethanol, or the like, while the reactions can be carried out both with an excess of compounds (3) and in the presence of bases, such as organic tertiary amines for example triethylamine, or pyridine. 12. The method of obtaining (2-oxo-1,2-dihydroquinolin-4-yl) aminocarboxylic acids and their derivatives of the general formula (1.3), where R is a hydrogen atom, n and Y have the above values according to claim 1, which consists in that 3-nitro-4-chloroquinolin-2 (1H) -one (10)

enter into interaction with aminocarboxylic acids or their derivatives of General formula (3)

, where n and Y have the above meanings in various solvents, such as benzene, toluene, ethyl acetate, dioxane, ethanol, or the like, while the reactions can be carried out both with an excess of compounds (3) and in the presence of bases, such as organic tertiary amines , for example triethylamine, or pyridine, also the interaction of compounds 3 and 10 can be carried out using pyridine as a solvent. 13. A method of treating epilepsy and paroxysmal conditions by administering effective amounts of the compounds of claims. 1-9.