RU2803571C1 - Application of 5-[(1h-indol-3-yl)methyl]-7-methyl-2,3-dihydro-1h-pyrrolo[1,2-d][1,4]diazepin-4(5h)-one as an antifungal agent for yeasts - Google Patents

Abstract

FIELD: organic chemistry.

SUBSTANCE: new biologically active compounds - substituted pyrrolo[1,2-d][1,4]diazepinones, namely 5-[(1 H-indol-3-yl)methyl]-7-methyl -2,3-dihydro-1 H-pyrrolo[1,2-d][1,4]diazepin-4(5 H )-one of Formula 1, which has antimycotic activity, which suggests its use in medicine as a potential drug with antimycotic properties. The use of 5-[(1H-indol-3-yl)methyl]-7-methyl-2,3-dihydro-1H-pyrrolo[1,2-d][1,4]diazepin-4(5H)-one as an antifungal agent against yeast fungi.

EFFECT: invention provides 5-[(1H-indol-3-yl)methyl]-7-methyl-2,3-dihydro-1H-pyrrolo[1,2-d][1,4]diazepin-4(5H)-one with 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 pyrrolo[1,2-d][1,4]diazepinones, namely 5-[(1H-indol-3-yl)methyl]-7-methyl- 2,3-dihydro-1H-pyrrolo[1,2-d][1,4]diazepin-4(5H)-one 1 formula:

compound 1 has antimycotic activity, which suggests its use in medicine as a potential 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 pyrrolo[1,2-d][1,4]diazepinones.

The goal is achieved by obtaining 5-[(1H-indol-3-yl)methyl]-7-methyl-2,3-dihydro-1H-pyrrolo[1,2-d][1,4]diazepine-4(5H) -one that has antimycotic activity.

The claimed compound 1 is synthesized by treating 5-methyl-2-(2'-aminoethyl)furan with N-Boc-tryptophan by boiling for 12 hours in absolute dichloroethane in the presence of 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide and N-methylmorpholine. Then the resulting crude amide is treated with a mixture of glacial acetic and concentrated hydrochloric acids at room temperature, followed by neutralization of hydrochloric acid Na ₂ CO ₃ , heating the reaction mixture at 120°C and isolating the target product using standard methods of synthetic organic chemistry according to the scheme:

Example 1. Preparation of compound 1. To a stirred solution of 912 mg (3 mmol) of N-Boc-tryptophan in 10 ml of absolute dichloroethane add 400 μl (3.2 mmol) of 5-methyl-2-(2'-aminoethyl)furan, 632 mg ( 3.3 mmol) 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide and 362 μl (3.3 mmol) N-methylmorpholine. The reaction mixture is refluxed for 12 hours. The solvent is then evaporated to dryness under reduced pressure. To a solution of the crude amide in ice-cold 22.5 ml of AcOH add 2.5 ml of conc. HCl. The reaction mixture is stirred at room temperature for 24 hours. Then 4.4 g of Na ₂ CO ₃ is added to the reaction mixture in portions and refluxed for 30 minutes. Next, the solvent is evaporated to dryness under reduced pressure. The residue was purified by column chromatography on silica gel using petroleum ether/ethyl acetate (9:1) as an eluent, purified product 1 was recrystallized from petroleum ether/ethyl acetate. Yield 58%. T _pl =201-202°C (with decomposition). Found, %: C, 73.43; N, 6.24; N, 14.68. C ₁₈ H ₁₉ N ₃ O. Calculated, %: C, 73.69; N, 6.53; N, 14.32. IR spectrum, (KBr), ν _max , cm ^-1 : 3281, 3178, 1649, 1257, 1094. ¹ H NMR spectrum, (400 MHz, DMSO-d ₆ ), d, ppm: 10.74 (br. s, 1H), 7.83 (br. s, 1H), 7.41-7.39 (m, 1H), 7.33-7.31 (m, 1H), 7.07-7.04 (m, 1H), 6.99-6.95 (m, 1H), 6.59 (s, 1H), 5.73 (d, J=3.0 Hz, 1H), 5.49 (d, J=3.0 Hz, 1H), 4.77-4.74 (m, 1H), 3.46-3.41 (m, 2H), 3.35 -3.32 (m, 1H), 3.17-3.09 (m, 2H), 2.96-2.87 (m, 1H), 1.46 (s, 3H). ¹³ C NMR spectrum (100 MHz, DMSO-d ₆ ), d, ppm: 170.4,135.9,130.3,128.8, 126.9, 123.8, 120.8, 118.4, 117.7, 111.2, 109.2, 105.5, 105.3, 62.8, 43.0 , 31.0, 26.6, 11.5; m/z (%) (EI, 70 eV) 293 (97) [M ⁺ ], 164 (57), 130 (100), 107 (60), 94 (17), 77 (12), 43 (64) . The resulting compound 1 is a light beige crystalline substance, soluble in ethanol, toluene, acetone, chloroform, and ethyl acetate.

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 LD ₅₀ is > 1500 mg/kg.

According to the classification of drug toxicity, compound 1 belongs to class V of practically non-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 possible the use of 5-[(1H-indol-3-yl)methyl]-7-methyl-2,3-dihydro-1H-pyrrolo[1,2-d][1,4]diazepine-4 (5H)-one 1 for the creation of new drugs as an antifungal agent against yeast fungi.

Claims (3)

Applications of 5-[( 1H- indol-3-yl)methyl]-7-methyl-2,3-dihydro- 1H- pyrrolo[1,2- d ][1,4]diazepine-4( 5H ) -she: as an antifungal agent against yeast fungi.