RU2806097C1 - ANTITUBERCULOSIS AGENT BASED ON 4'-HYDROXY-1'-(2-HYDROXYPHENYL)-3'-ACYL-SPIRO[BENZO[b][1,4]THIAZINE-2,2'-PYRROLE]-3,5'(1 'H,4H)-DIONES - Google Patents

Abstract

FIELD: organic chemistry; pharmacology.

SUBSTANCE: use of 4'-hydroxy-1'-(2-hydroxyphenyl)-3'-acyl-spiro[benzo[b][1,4]thiazine-2,2'-pyrrole] -3,5'(1'H, 4H)-diones of the general formula:

where R = 2-furyl (a), 4-C₆H₄NO₂ (b) as an anti-tuberculosis agent. The above compounds have been shown to have anti-tuberculosis activity.

EFFECT: development of a drug with anti-tuberculosis activity, which can be used in pharmacology as a potential drug with anti-tuberculosis properties.

1 cl, 1 tbl, 3 ex

Description

The invention relates to the field of organic chemistry, namely to biologically active 4'-hydroxy-1'-(2-hydroxyphenyl)-3'-acyl-spiro[benzo[b][1,4]thiazine-2,2'-pyrrole ]-3,5'(1'H,4H)-diones of the formula:

where R=2-furyl (a), 4-C ₆ H ₄ NO ₂ (b),

possessing antituberculosis activity, which suggests their use in pharmacology and medicine as drugs with an antituberculosis effect.

Structural analogues of the claimed compounds are known - derivatives containing a fragment of 1,5-dihydro-2H-pyrrol-2-one, which have high anti-tuberculosis activity in vitro (Intaraudom S., Boonyuen N., Suvannakad R., Rachtawee P., Pittayakhajonwut P., Penicolinates A-E from endophytic Penicillium sp.BCC16054. // Tetrahedron Letters. - 2013. - T. 54. P. 744-748).

The anti-tuberculosis drug pyrazinamide [Y. Zhang, W.W. Yew. Mechanisms of development of drug resistance in Mycobacterium tuberculosis / International Journal of Tuberculosis and Lung Diseases. T 2. No. 1, 2011, pp. 7-19], widely used in medical practice.

The objective of the invention is to search in a series of derivatives for 4'-hydroxy-1'-(2-hydroxyphenyl)-3'-acyl-spiro[benzo[b][1,4]thiazine-2,2'-pyrrole]-3,5 '(1'H,4H)-diones of substances with a pronounced anti-tuberculosis effect and expanding the arsenal of means of influencing a living organism.

This goal is achieved by obtaining 4'-hydroxy-1'-(2-hydroxyphenyl)-3'-acyl-spiro[benzo[b][1,4]thiazine-2,2'-pyrrole]-3,5'(1 'H,4H)-diones, which have anti-tuberculosis activity.

The claimed compounds are synthesized by reacting 3-(het)aroyl-2,4-dihydro-1H-pyrrolo[2,1-c][1,4]benzoxazine-1,2,4-triones (I a,b) with o -aminothiophenol in a ratio of 1:1.5 in an anhydrous solvent to isolate the target products (IIa, b) according to the presented scheme:

where R=2-furyl (a), 4-C ₆ H ₄ NO ₂ (b).

The process is carried out at t=25°C and anhydrous acetonitrile is used as a solvent.

The invention is illustrated by the following examples.

Example 1. Preparation of 3'-(furan-2-carbonyl)-4'-hydroxy-1'-(2-hydroxyphenyl)spiro[benzo[b][1,4]thiazine-2,2'-pyrrole]-3 ,5'(1H,4H)-dione (IIa).

To a solution of 1.0 mmol 3-(furan-2-carbonyl)-1H-benzo[b]pyrrolo[1,2-d][1,4]oxazine-1,2,4-trione (Ia) in 2 ml of anhydrous acetonitrile 1.5 mmol o-aminothiophenol was added. The reaction mass was stirred at t=25°C for 10 minutes, the resulting precipitate was filtered off. Yield 78%, mp. 224-226°C. Compound (IIa) C ₂₂ H ₁₄ N ₂ O ₆ S.

Found, %: C 60.41; H 3.29; N 6.37; S 7.51.

Calculated, %: C 60.83; And 3.25; N 6.45; S 7.38.

Compound (IIa) is a light yellow crystalline substance, readily soluble in DMSO, DMF, soluble in dichloromethane, chloroform, acetone, alcohols, insoluble in water and alkanes. Stable when stored under normal conditions

The IR spectrum of compound (IIa), recorded as a paste in petroleum jelly, contains bands of stretching vibrations of hydroxo groups 3363, 3292 cm ^-1 , lactam NH group 2925 cm ^-1 , carbonyl groups of lactam, aroyl fragments and pyrrole ring 1717, 1661, 1625 cm ^-1 .

^1H NMR spectrum (400 MHz, DMSO-d ₆ , δ, ppm): 10.89 s (1H), 9.62 s (1H), 7.99 d (J=1.6 Hz, 1H), 759 d (J =3.6 Hz, 1H), 7.07-6.98 m (2H), 6.95-6.86 m (2H), 6.80-6.69 m (4H), 6.60 td (J=7.6, 1.4 Hz, 1H). One of the OH groups is invisible in the ¹ H NMR spectrum.

Example 2. Preparation of 4'-hydroxy-1'-(2-hydroxyphenyl)-3'-(4-nitrobenzoyl)spiro[benzo[b][1,4]thiazine-2,2'-pyrrole]-3,5 '(1'H,4H)-dione (IIb).

1.5 mmol o-aminothiophenol. The reaction mass was stirred at t=25°C for 10 minutes, the resulting precipitate was filtered off. Yield 70%, mp. 236-238°C. Compound (IIb) C ₂₄ H ₁₅ N ₃ O ₇ S.

Found, %: C 58.13; II 3.14; N 8.73; S 6.44

Calculated, %: C 58.89; H 3.09; N 8.59; S 6.55

Compound (IIb) is a yellow crystalline substance, readily soluble in DMSO, DMF, soluble in dichloromethane, chloroform, acetone, alcohols, insoluble in water and alkanes. Stable when stored under normal conditions

The IR spectrum of compound (IIb), recorded as a paste in petrolatum oil, contains bands of stretching vibrations of hydroxo groups 3384, 3303 cm ^-1 , lactam NH group 2921 cm ^-1 , carbonyl groups of lactam and aroyl fragments and pyrrole ring 1726, 1667, 1644 cm ^-1 .

^1H NMR spectrum (400 MHz, DMSO-d ₆ , δ, ppm): 10.94 s (1H), 9.59 s (1H), 8.34 d (J=8.8 Hz, 2H), 7.93 d (J=8.7 Hz, 2H), 7.05-6.95 m (3H), 6.91 td (J=7.7, 1.4 Hz, 1H), 6.71-6.79 m (3H), 6.57 td (J=7.7, 1.4 Hz, 1H). One of the OH groups is invisible in the ¹ H NMR spectrum.

Example 3. Pharmacological study of 4'-hydroxy-1'-(2-hydroxyphenyl)-3'-acyl-spiro[benzo[b][1,4]thiazine-2,2'-pyrrole]-3,5'( 1'H,4H)-diones (IIa,b) for the presence of anti-tuberculosis activity.

Assessment of the tuberculostatic activity of the test substance was carried out by the method of two-fold serial dilutions in a liquid nutrient medium using the growth indicator resazurin sodium salt [Resazurin Microliter Assay Plate: Simple and Inexpensive Method for Detection of Drag Resistance in Mycobacterium tuberculosis / J. - C. Palomino, A. Martin , M. Camacho, et al. // Antimicrobial Agents and Chemotherapy. - 2002. V. 46, No. 8. - P. 2720-2722].

The strain of M. tuberculosis H37Rv ATCC 25618, obtained from the State Scientific Center for Medical and Biology “State Collection of Pathogenic Microorganisms and Cell Cultures”, was used as a test microorganism.

In the wells of a sterile 96-well flat-bottomed microplate, two parallel rows of two-fold serial dilutions of the claimed compound were prepared in Middlebrook nutrient liquid medium. 7H, containing the enrichment additive VASTES MGIT OADC (oleic acid, albumin, dextrose and catalase). Each well contained 150 μl of a certain concentration of the test substance.

A bacterial suspension was prepared from 21 day-old cultures of M. tuberculosis H37Rv, which was standardized according to the Farland optical turbidity standard No. 1 using a densitometer, after which the inoculum was diluted 1:20 in Middlebrook 7H9 nutrient medium and 150 μl was added to each well of the plate.

The last rows contained nutrient medium and culture in equal volumes (control). The maximum concentration tested was 1000 μg/ml, the minimum was 1 μg/ml.

The plates were sealed in plastic bags and incubated at 37°C for 8 days. Then 30 μl of a 0.01% sodium resazurinate solution prepared in sterile distilled water was added to each well and cultured for another 24 hours at 37°C, then the color change of resazurin was visually assessed. A change from blue to pink indicates a decrease in resazurin and, therefore, growth of mycobacteria. MIC is the minimum inhibitory concentration of chemicals at which no color change occurs.

Analysis of the data obtained showed that compounds IIa, IIb have a tuberculostatic effect on the test culture at concentrations of 62.5-250 μg/ml.

The claimed 4'-hydroxy-1'-(2-hydroxyphenyl)-3'-acyl-spiro[benzo[b][1,4]thiazine-2,2'-pyrrole]-3,5'(1'H, 4H)-diones can find application in pharmacology as potential drugs with antituberculosis properties.

Claims (3)

Application of 4'-hydroxy-1'-(2-hydroxyphenyl)-3'-acyl-spiro[benzo[ b ][1,4]thiazine-2,2'-pyrrole]-3,5'( 1'H , 4 H )-diones of the general formula: , where R = 2-furyl (a), 4-C ₆ H ₄ NO ₂ (b) as an anti-tuberculosis agent.