RU2803600C1 - APPLICATION OF 10-METHYL-6,7-DIHYDRO-5H-PYRIDO[2,3-b]-PYRROLO[1,2-d][1,4]DIAZEPINE AS AN ANTIFUNGAL AGENT AGAINST YEASTS - Google Patents

Abstract

FIELD: organic chemistry.

SUBSTANCE: new biologically active compounds - substituted pyrido[b]pyrrolo[1,2-d][1,4]diazepines, namely 10-methyl-6,7-dihydro-5H-pyrido[2 ,3-b]pyrrolo[1,2-d][1,4]diazepine of Formula 1, which has antimycotic activity, which suggests its use in medicine as a drug with antimycotic properties. The use of 10-methyl-6,7-dihydro-5H-pyrido[2,3-b]pyrrolo[1,2-d][1,4]diazepine of Formula (1) as an antifungal agent against yeast fungi is presented.

EFFECT: invention provides 10-methyl-6,7-dihydro-5H-pyrido[2,3- b]pyrrolo[1,2-d][1,4]diazepine of Formula (1), which has a pronounced antimycotic effect and low toxicity.

1 cl, 1 tbl, 3 ex

Description

The invention relates to the field of organic chemistry - new biologically active compounds - substituted pyrido[b]pyrrolo[1,2-d][1,4]diazepines, namely 10-methyl-6,7-dihydro-5H-pyrido[2 ,3-b]pyrrolo[1,2-d][1,4]diazepine 1 formula:

compound 1 has antimycotic activity, which suggests its use in medicine as a drug with antimycotic properties.

The structural analogue of the claimed compound is 7-chloro-4-(4-ethylphenyl)-5,6-dihydro-4H-benzo[f]pyrrolo[1,2-a][1,4]diazepine 2, which has antimycotic activity [ Bioorg. Med. Chem. Lett. 2005, 15, 3453 - 3458; doi: 10.1016/j.bmcl.2005.05.007] formulas:

Ketoconazole 3 formulas were chosen as the standard of comparison:

which is widely used in medical practice and is analogous in action [Mashkovsky M.D. Medicines. - 16th ed., revised, corrected. and additional - M.: New Wave, 2012. - p. 918; Med. Chem. Res. 2013, 22, 211 -218; doi:10.1007/s00044-012-0021-2].

The objective of the invention is to search for substances with a pronounced antimycotic effect and low toxicity in the series of substituted pyrido[b]pyrrolo[1,2-d][1,4]diazepines.

This goal is achieved by obtaining 10-methyl-6,7-dihydro-5H-pyrido[2,3-b]pyrrolo[1,2-d][1,4]diazepine, which has antimycotic activity.

The claimed compound 1 is synthesized by a two-step method, which includes treatment of N-[2-(5-methylfuran-2-yl)ethyl]-3-nitropyridin-2-amine 4 with a mixture of glacial acetic and concentrated hydrochloric acids at room temperature and treatment of the resulting 7- [(3-nitropyridin-2-yl)amino]heptan-2,5-dione 5 with carbonyl iron powder in glacial acetic acid at boiling, followed by isolation of the target product using standard methods of synthetic organic chemistry according to the scheme:

Example 1. Preparation of compound 1. Step 1. To a solution of 741 mg (3 mmol) N-[2-(5-methylfuran-2-yl)ethyl]-3-nitropyridin-2-amine 4 in 16 ml of glacial acetic acid is added concentrated hydrochloric acid (12 M, 3.2 ml). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was then poured into H ₂ O (50 ml) and neutralized to pH ~ 7 with NaHCO ₃ . The product is extracted with ethyl acetate (3×30 ml). The combined organic fractions are washed with saturated NaCl solution (3×20 ml), dried over anhydrous Na ₂ SO ₄ . The solvent is evaporated to dryness under reduced pressure. The residue was purified by silica gel column chromatography using petroleum ether/ethyl acetate (4:1) as the eluent to give purified product 5 as a yellow oil. Yield 80%. Step 2. To a solution of 398 mg (1.5 mmol) of 7-[(3-nitropyridin-2-yl)amino]heptan-2,5-dione 5 in glacial acetic acid (6 ml) add 1.26 g (22.5 mmol) of carbonyl iron . The reaction mixture is refluxed for 15 minutes. The reaction mixture is then poured into H ₂ O (100 ml) and neutralized to pH ~ 7 with NaHCO ₃ . Unreacted iron is filtered off and washed with hot ethyl acetate (3×5 ml). The aqueous layer was extracted with ethyl acetate (3×40 ml). The combined organic fractions are washed with saturated NaCl solution (3×20 ml), dried over anhydrous Na ₂ S0 ₄ . The solvent is evaporated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel using petroleum ether/ethyl acetate (10:1) as an eluent. Product 1 is recrystallized from petroleum ether/ethyl acetate. Yield 84%. T _pl = 151-152°C. Found, %: C, 72.33; N, 6.49; N, 21.07. _{C12H13N3 . _}

Calculated, %: C, 72.33; N, 6.58; N, 21.09. IR spectrum, (CaF ₂ ), ν _max , cm ^-1 : 3232, 1595, 1523, 1410, 1325, 1304, 1242, 1220. ¹ H NMR spectrum, (400 MHz, DMSO-d ₆ ), d, m. d.: 7.98-7.99 (m, 1H), 7.46 (d, J=7.6 Hz, 1H), 6.80-6.77 (m, 1H), 6.08 (br. s, 1H), 5.88 (d, J=3.2 Hz , 1H), 5.83 (d, J=3.2 Hz, 1H), 3.51-3.47 (m, 2H), 2.76-2.73 (m, 2H), 2.14 (s, 3H). ¹³ C NMR spectrum (100 MHz, DMSO-d ₆ ), d, ppm: 152.9, 145.4, 133.4, 132.4, 127.5, 120.5, 113.6, 108.4, 103.7, 49.6, 25.1, 13.2. The resulting compound 1 is a pale yellow crystalline substance, soluble in chloroform, toluene, acetone, and ethanol.

Example 2. To characterize the antimycotic activity of compounds, standard parameters were used: minimum inhibitory concentration (MIC), which was determined by a modified method of two-fold serial dilutions (Guidelines: Fungi of the genus Candida. Methods of isolation, identification at the species level and determination of sensitivity to antifungal drugs, Moscow, 2009; MUK 4.2.1890-04 Determination of the sensitivity of microorganisms to antibacterial drugs) and minimum fungicidal concentration (MFC).

Tests were performed using a culture of model microorganisms Candida albicans ATCC 10231 on Mueller-Hinton nutrient medium supplemented with 2% glucose in 96-well polystyrene plates. The concentration of microorganisms in the wells before the start of cultivation was 2.5*10 ⁵ CFU/ml. Cultivation was carried out at 37°C without stirring. MIC determination and inoculations to determine MPA were carried out after 24 hours. The test compounds were dissolved in dimethyl sulfoxide (DMSO) until complete dissolution at a concentration of 20 mg/ml and lower, depending on solubility. Added to the nutrient medium so that the amount of DMSO did not exceed 5%.

Example 3. Acute toxicity (LD ₅₀ , mg/ml) of compound 1 was determined according to the method of G.N. Pershina [Pershin G.N. Methods of experimental chemotherapy // M., S. 100, 1971, 109-117]. Compound 1 was administered intraperitoneally to white mice weighing 16-18 g in the form of a suspension in 2% starch mucus, and the behavior and death of the animals were observed for 10 days. For test compound 1, the _LD50 is >500 mg/kg.

According to the classification of drug toxicity, compound 1 belongs to class IV of low-toxic drugs [Izmerov N.F., Sanotsky I.V., Sidorov K.K. Parameters of toxicometry of industrial poisons with a single exposure: Handbook. M., 1977, p. 196]. The test results are presented in the table:

As can be seen from the table, the claimed compound 1 is comparable in antimycotic activity to the reference drug (Ketoconazole) against C. albicans. Thus, this makes it possible to use 10-methyl-6,7-dihydro-5H-pyrido[2,3-b]pyrrolo[1,2-d][1,4]diazepine 1 to create new drugs as an antifungal agents against yeast fungi.

Claims (3)

Use of 10-methyl-6,7-dihydro- 5H -pyrido[2,3- b ]pyrrolo[1,2- d ][1,4]diazepine: as an antifungal agent against yeast fungi.