RU2836731C1 - 1'-(3,4-dichlorophenyl)-2-oxo-5',6',7',7a'-tetrahydrospiro[indoline-3,3'-pyrrolysine]-2',2'(1'h)- dicarbonitrile with antimicrobial properties - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as an agent with antimicrobial activity. Technical result is achieved due to use of 1'-(3,4-dichlorophenyl)-2-oxo-5',6',7',7a'-tetrahydrospiro[indoline-3,3'-pyrrolysine]-2',2'(1'H)-dicarbonitrile of formula (I) or a pharmacologically acceptable salt thereof as an agent with antimicrobial activity for controlling human and animal diseases caused by strains of bacteria Pseudomonas aeruginosa, or Escherichia coli, or Staphylococcus aureus.

EFFECT: wider range of pharmaceutical agents.

1 cl, 1 tbl

Description

Field of technology

The invention relates to the field of pharmaceutical chemistry, namely to a compound of formula ( I ) or its pharmaceutically acceptable salt, and can be used in medicine and veterinary science.

( I )

State of the art

A topical issue in modern pharmaceutical chemistry remains the search for simple and environmentally friendly methods for synthesizing new antimicrobial compounds. Among these, spiro-fused indole derivatives (1-3) and methods for synthesizing these compounds continue to be actively studied.

Spirothiapyrrolizidines ( 1a-d ) and spiropyrrolidines ( 2a-h ) are known, containing cyano groups as substituents in the pyrrolidine ring and various substituents in the indole and phenyl fragments (1). Formulas:

The antimicrobial activity of the above compounds was tested against gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and gram-positive bacteria Staphylococcus aureus . The minimum inhibitory concentration (MIC) was determined. Gentamicin and Ampicillin were used as comparison drugs. Among the studied bacteria, good activity was demonstrated only against Klebsiella pneumoniae , with the best results demonstrated by indole derivatives containing the spirothiazolidine fragment 1b-d . The MIC of spirothiapyrrolizidine 1c is 0.005 mg/ml, which is 156 and 78 times higher than the values of the standards Gentamicin and Ampicillin, respectively.

Spiropyrrolidines ( 3a-e ) are known that contain cyano groups as substituents in the pyrrolidine ring and various heteroaromatic substituents in the 4 position of the pyrrolidine cycle (2). Formulas:

The antimicrobial activity of the above compounds was tested against gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa , Klebsiella pneumoniae, Acinetobacter baumannii and gram-positive bacteria Staphylococcus aureus using the agarose gel diffusion method. Levofloxacin was used as a comparison drug. The activity of the studied compounds is much lower than the activity of the Levofloxacin standard.

1'-(3,4-Dichlorophenyl)-2-oxo-5',6',7',7a'-tetrahydrospiro[indoline-3,3'-pyrrolizine]-2',2'(1'H)-dicarbonitrile was synthesized (3), but its antimicrobial properties have not been studied.

Disclosure of the essence of the invention

The technical challenge is to expand the class of substances with antimicrobial properties suitable for use in medicine and veterinary medicine.

The technical result consists in expanding the arsenal of pharmaceutical agents due to the representative of substituted spiropyrrolizidine dicarbonitriles.

The technical result is achieved by using 1'-(3,4-dichlorophenyl)-2-oxo-5',6',7',7a'-tetrahydrospiro[indoline-3,3'-pyrrolizine]-2',2'(1'H)-dicarbonitrile of formula ( I )

(I)

or its pharmacologically acceptable salt as an agent with antimicrobial activity for combating diseases of humans and animals caused by strains of bacteria Pseudomonas aeruginosa , or Escherichia coli , or Staphylococcus aureus .

Implementation of the invention

The antimicrobial properties of 1'-(3,4-dichlorophenyl)-2-oxo-5',6',7',7a'-tetrahydrospiro[indoline-3,3'-pyrrolizine]-2',2'(1'H)-dicarbonitrile I were studied.

A three-component synthesis of indole 1'-(3,4-dichlorophenyl)-2-oxo-5',6',7',7a'-tetrahydrospiro[indoline-3,3'-pyrrolizine]-2',2'(1'H)-dicarbonitrile I was carried out using a simple, environmentally friendly method in one step, starting from 2-(3,4-dichlorobenzylidene)malononitrile, isatin and proline, using isopropyl alcohol as a solvent (3).

To evaluate the antimicrobial activity of the synthesized compound, the half-maximum effective concentration (EC ₅₀ ) was determined in comparison with the drug "Ciprofloxacin" (OJSC "Sintez", Russia, Kurgan). Antimicrobial activity was assessed against Staphylococcus aureus ( Staphylococcus aureus B-8172), Escherichia coli ( Escherichia coli K-12) and Pseudomonas aeruginosa ( Pseudomonas aeruginosa B-6643), obtained from the All-Russian Collection of Industrial Microorganisms. The studies were carried out using the method of measuring the optical density of bacterial cultures during growth on a liquid nutrient medium LB containing yeast extract (5 g / l), tryptone (10 g / l) and sodium chloride (5 g / l).

Table 1

EC ₅₀ , μg/ml
Bacterial strains P. aeruginosa
ATCC 9027 S. aureus
ATCC 25923 E. coli K-12 Compound I 5.9±1.1 3.4±0.3 80±8 Ciprofloxacin 69±7 36±4 0.56±0.16

Compound I demonstrated antimicrobial activity against Pseudomonas aeruginosa that was 11 times greater than the commercial drug Ciprofloxacin, and 10 times greater against Staphylococcus aureus . Weaker antimicrobial activity was detected against Escherichia coli , being more than 140 times less than that of Ciprofloxacin.

Methodology for assessing antimicrobial activity

The bacterial culture was inoculated onto liquid LB medium for 18 hours. Then 100 μl of the culture was added to 25 ml of liquid LB medium and incubated at 37°C for 1 hour. 150 μl of liquid LB nutrient medium were added to each well of a 96-well microplate. 24 μl of the test compound and comparison substance solutions in DMSO at a concentration of 10 mg/ml were added to the first wells, they were diluted twofold from 400 to 0.195 μg/ml, and 150 μl of the 1-hour bacterial culture containing approximately 10 ⁵ cells per 1 ml were added to each well of the plate.

The optical density was measured in each well at 0 hours of incubation using a Multiskan Ascent plate reader (Thermo Scientific, Finland) at a wavelength of 595 nm. The plates were then incubated in a shaker-incubator for 1-8 hours at 37°C. The optical density was measured every hour at 595 nm in each well of the plate. The obtained data were processed using the Excel 2007 software package (Microsoft, USA), and the EC ₅₀ value of the compounds (μg/ml) was determined based on the graph.

All experiments to determine antimicrobial activity were performed in triplicate. Statistical analysis of the data was performed using the Excel 2007 software package (Microsoft, USA). The results are presented as mean values with confidence intervals at P = 0.05 (confidence level 95%).

Literature

1. Dandia A., Khan S., Soni P., Indora A., Mahawar D., Pandya P., Chauhan C. S. Diversity-oriented sustainable synthesis of antimicrobial spiropyrrolidine/thiapyrrolizidine oxindole derivatives: New ligands for a metallo-b-lactamase from Klebsiella pneumonia // Bioorganic & Medicinal Chemistry Letters. 2017. Vol. 16. No. 27. P. 2873-2880. DOI: 10.1016/j.bmcl.2017.04.083.

2. Vidya S., Priya K., Jayasree V., Deepthi A., Biju P. Synthesis of heterocycle appended spiro(oxindole-3,2'-pyrrolidine) derivatives from heterocyclic ylidenes and azomethine ylide through 1,3-dipolar cycloaddition reactions//Synthetic Communications. 2019. Vol. 49. No. 12. P.1592-1602. DOI: 10.1080/00397911.2019.1605444.

3. Borisova S.B. Reactions of 1,3-dipolar cycloaddition in the synthesis of spiro-fused pyrrolidines and pyrrolizidines. Diss. for the degree of candidate of chemical sciences, Saratov, 2023. P.151.

Claims (3)

Use of the compound 1'-(3,4-dichlorophenyl)-2-oxo-5',6',7',7a'-tetrahydrospiro[indoline-3,3'-pyrrolizine]-2',2'(1'H)-dicarbonitrile of formula ( I ) ( I ) or a pharmaceutically acceptable salt thereof as an agent having antimicrobial activity for combating diseases in humans and animals caused by strains of bacteria Pseudomonas aeruginosa , or Escherichia coli , or Staphylococcus aureus .