RU2790379C1 - The use of 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-2-thioxo-1,3,6-triazaspiro[4.4]non-8-en-4,7-diones as agents with antimicrobial activity against s. aureus - Google Patents

Abstract

FIELD: microbiology.

SUBSTANCE: invention relates to the use of 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-2-thioxo-1,3,6-triazaspiro[4.4]non-8-en-4,7-diones of the general formula

,

where: Ar=4-ClC6H4, R1=H, R2=B u (a); Ar=4-ClC6H4, R1=Me, R2=Ph (b), as agents with antimicrobial activity against S. aureus.

EFFECT: antimicrobial activity against S. aureus compounds of class 2-thioxo-1,3,6-triazaspiro[4.4]non-8-en-4,7-diones.

1 cl, 2 tbl, 3 ex

Description

The invention relates to the field of organic chemistry, namely the use of individual compounds of the class 2-thioxo-1,3,6-triazaspiro[4.4]non-8-ene-4,7-diones as agents with antimicrobial activity, which can be used as initial products for the synthesis of new heterocyclic systems and in pharmacology.

The claimed compounds 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-2-thioxo-1,3,6-triazaspiro[4.4]non-8-ene-4,7-diones and the method of their synthesis are known from the level technology. The claimed compounds are products of the interaction of 8-aroyl-3,4-dihydro-1H-pyrrolo[2,1-c][1,4]oxazine-1,6,7-triones with substituted thioureas (Facile regiodivergent synthesis of spiro pyrrole- substituted pseudothiohydantoins and thiohydantoins via reaction of [e]-fused 1H-pyrrole-2,3-diones with thiourea Kobelev A.I., Tretyakov N.A., Stepanova E.E., Dmitriev M.V., Rubin M., Maslivets A.N. Beilstein JOC, 2019, 15, 2864 2871. doi: 10.3762/bjoc.15.280), formed as follows:

where: 1: Ar=4-ClC ₆ H ₄ (a), 4-CH ₃ C ₆ H ₄ (b); 2: R ¹ =H (a), Ph (b); 3: Ar=4-ClC ₆ H ₄ , R ¹ =H (a), Ph (b); Ar=4-CH ₃ C ₆ H ₄ , R ¹ =H (c), Ph (d).

The antimicrobial activity of 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-2-thioxo-1,3,6-triazaspiro[4.4]non-8-en-4,7-diones was previously studied (Pat. 2763731 RF, IPC A61K 31/407 Use of 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-1,3-dialkyl-2-thioxo-1,3,6-triazaspiro[4.4]non-8-ene -4,7-diones as agents with antimicrobial activity / N. A. Tretyakov, S. Yu. Balandina, A. N. Maslivets, applicants and patent holder Perm State National Research University (RU) 2021118805; Application 06/17/2021; published 12/30/2021, Bulletin No. 1.) (Table 1):

The objective of the invention is to find new compounds with a higher antimicrobial activity, and to expand the arsenal of means of influencing a living organism.

EFFECT: previously unknown antimicrobial activity against S. aureus 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-2-thioxo-1,3,6-triazaspiro[4.4]non-8-en- 4,7-diones.

The problem is solved by the fact that the compounds 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-2-thioxo-1,3,6-triazaspiro[4.4]non-8-ene-4,7-diones exhibit a high antimicrobial activity against S. aureus at a concentration of 250 µg/ml.

The claimed compounds are synthesized by reacting 8-aroyl-3,4-dihydro-1H-pyrrolo[2,1-c][1,4]oxazine-1,6,7-triones with substituted thioureas in a solvent medium, followed by isolation of target products , according to the following scheme:

where: 4: Ar=4-ClC ₆ H ₄ , R ¹ =H (a), Me (b); 5: R ² =Bu (a), Ph (b); 6: Ar=4-ClC ₆ H ₄ , R ¹ =H, R ² =Bu (a); Ar \u003d 4-ClC ₆ H ₄ , R ¹ \u003d Me, R ² \u003d Ph (b),

The process is carried out at a temperature of 108-110°C, and toluene is used as a solvent.

The invention is illustrated by the following examples.

Example 1 1,3-Dibutyl-8-hydroxy-6-(2-hydroxyethyl)-2-thioxo-9-(4-chlorobenzoyl)-1,3,6-triazaspiro[4,4]non-8-ene -4,7-dione (6a).

0.631 mmol N,N-dibutylthiourea, boiled for 5 h, the precipitate was filtered off. Yield 76%, m.p. 81-83°C (toluene).

Compound (6а) C ₂₃ H ₂₈ ClN ₃ O ₅ S.

Found, %: С 56.27; H 5.39; N 8.18

Calculated, %: С 55.92; H 5.71; N 8.51.

Compound (6a) is a light yellow crystalline substance, readily soluble in DMSO and DMF, sparingly soluble in less polar organic solvents, and insoluble in water and alkanes. Stable when stored under normal conditions.

In the IR spectrum of compound (6а), recorded as a paste in vaseline oil, there are bands of stretching vibrations of the alcohol group OH in the form of a broad peak at 3512 cm ^-1 , the enol group OH in the form of a wide band at 3348 cm ^-1 , two lactam carbonyl groups at 1738 cm ^-1 and 1732 cm ^-1 respectively, the thioxo group C=S at 1662 cm ^-1 and the aroyl carbonyl group at 1632 cm ^-1 .

In the ^1H NMR spectrum of compound (6a), recorded in a solution in DMSO-d ₆ , in addition to the signals of aromatic ring protons, there are two triplets of methyl groups of butyl substituents at 0.70 ppm. and 0.94 ppm, a group of methylene protons of butyl substituents at 1.11-1.24 ppm. (m, 3H), 1.34-1.43 (m, 2H), 1.54-1.63 (m, 1H), 1.65-1.72 (m, 2H), 3.08-3.11 (m, 2H) and 3.80-3.88 (m, 2H) , a group of 4 methylene protons of the ethoxy substituent at 3.21-3.28 ppm (m, 1H), 3.32-3.38 ppm (m, 1H), 3.46-3.52 ppm (m, 1H), 3.62–3.68 (m, 1H), respectively, and a broadened singlet of the proton of the OH ethoxy group at 5.10 ppm.

Example 2 8-Hydroxy-6-(2-hydroxypropyl)-2-thioxo-1,3-diphenyl-9-(4-chlorobenzoyl)-1,3,6-triazaspiro[4,4]non-8-ene -4,7-dione (6b).

To a solution of 0.560 mmol 3-methyl-8-(4-chlorobenzoyl)-3,4-dihydro-1H-pyrrolo[2,1-c][1,4]oxazine-1,6,7-trione in 15 ml of anhydrous toluene, 0.560 mmol of N,N-diphenylthiourea was added, the mixture was boiled for 5 h, and the precipitate was filtered off. Yield 69%, m.p. 264-265°C (toluene).

Compound (6b) C ₂₈ H ₂₂ ClN ₃ O ₅ S.

Found, %: С 61.72; H 3.71; No. 7.38

Calculated, %: С 61.37; H 4.05; N 7.67.

Compound (6b) is a light yellow crystalline substance, readily soluble in DMSO and DMF, sparingly soluble in less polar organic solvents, and insoluble in water and alkanes. Stable when stored under normal conditions.

In the IR spectrum of compound (6b), recorded as a paste in vaseline oil, there are bands of stretching vibrations of the alcohol group OH in the form of a broad peak at 3617 cm ^-1 , the enol group OH in the form of a wide band at 3173 cm ^-1 , two lactam carbonyl groups at 1731 cm ^-1 and 1717 cm ^-1 respectively, the thioxo group C=S at 1674 cm ^-1 and the aroyl carbonyl group at 1627 cm ^-1 .

In the ^1H NMR spectrum of compound (6а), recorded in a solution in DMSO-d ₆ , in addition to the signals of protons of aromatic rings, there is a doublet of the methyl group of the hydroxypropyl fragment at 1.13 ppm, two doublets of doublets of the methylene group of the hydroxypropyl substituent at 3.41 ppm . and 3.72 ppm, a multiplet of the methane group of the hydroxypropyl substituent at 3.79-3.88 ppm, and a broadened singlet of the proton of the ethoxy OH group at 5.68 ppm.

Example 3. Pharmacological study of compounds (6a,b) for the presence of antimicrobial activity.

For research, the generally accepted method of two-fold serial dilutions in a liquid nutrient medium by the micromethod was used [Guidelines for the experimental (preclinical) study of new pharmacological substances - M.: I-vo Medicine, 2005]. Initial dilutions of microorganisms in physiological saline were prepared from daily agar culture according to the optical turbidity standard (OCO) for 5 ME using a densitometer. After a series of dilutions, the final concentration of cells in the experiment was 2.5×10 ⁵ cells/ml.

The antimicrobial properties of the chemical were studied on 3 collection opportunistic strains of microorganisms: Staphylococcus aureus (strain 906), Escherichia coli (strain 1257), Candida albicans (RKPGU 1353/1277), obtained at the Scientific Center for Expertise of Medicinal Products Ministry of Health and Social Development of Russia. The fact of inhibition (inhibition of growth) of microbial cells in dilutions of preparations was noted after 20 hours of thermostating at 37°C. The final results were recorded 7 days after inoculation on RPA agar slant. The maximum tested concentration of the compounds corresponded to 1000.0 μg/ml. Antimicrobial (inhibitory, bactericidal) activity was evaluated by the minimum effective concentration.

Analysis of the obtained data showed:

• ; compounds 6a, b have antimicrobial activity against the culture of S. aureus; inhibitory effect at a concentration of 250.0 µg/ml;

The proposed substances 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-2-thioxo-1,3,6-triazaspiro[4.4]non-8-en-4,7-diones (6a,b) have a pharmacological activity, namely antimicrobial activity against S. aureus , and can be used in pharmacology as potential drugs.

Claims (4)

Application of 9-aroyl-8-hydroxy-6-(2-hydroxyalkyl)-2-thioxo-1,3,6-triazaspiro[4.4]non-8-en-4,7-diones of the general formula

, where: Ar=4-ClC ₆ H ₄ , R ¹ =H, R ² =Bu (a); Ar \u003d 4-ClC ₆ H ₄ , R ¹ \u003d Me, R ² \u003d Ph (b), as agents with antimicrobial activity against S. aureus.