RU2781135C1 - Benzylammonium salt of 4-ethoxycarbonyl-5-oxo-2-(2-oxo-2-phenylethylidene)-2,5-dihydro-1h-pyrrole-3-olate, showing antimicrobial activity - Google Patents

Abstract

FIELD: benzyl ammonium salts.

SUBSTANCE: invention relates to the benzyl ammonium salt of 4-ethoxycarbonyl-5-oxo-2-(2-oxo-2-phenylethylidene)-2,5-dihydro-1H-pyrrol-3-olate of the formula I.

EFFECT: obtaining a new compound, which can be used in medicine as an antimicrobial drug.

1 cl, 1 tbl

Description

The claimed compound belongs to the field of organic chemistry, to derivatives of 5-oxopyrroles, namely to new biologically active substances - benzyl ammonium salt of 4-ethoxycarbonyl-5-oxo-2- (2-oxo-2-phenylethylidene) -2,5-dihydro- 1H-pyrrole-3-olate formula:

which can be used as an antimicrobial drug.

Compound I is prepared by reacting 2-amino-5-(2-oxo-2-phenylethylidene)-4-oxo-1H-4,5-dihydrofuran-3-carboxylic acid ethyl ester II with benzylamine in ethanol. This compound is a yellow crystalline substance, soluble in dimethylformamide, dimethyl sulfoxide, when heated - in ethanol, stable during storage. Structural analogues of the claimed compound are known, obtained from N-acylhomoserine lactones by the irreversible Claisen reaction, having antimicrobial, antiviral, fungicidal activity, as well as antitumor properties [Brodyck J.L. Royles. Naturally Occurring Tetramic Acids: Structure, Isolation, and Synthesis // Chem. Rev. - 1995. - V. 95, No. 6. - P. 1981-2001; Colin A. Lowery, Junguk Park, Christian Gloeckner [et al.]. Defining the Mode of Action of Tetramic Acid Antibacterials Derived from Pseudomonas aeruginosa Quorum Sensing Signals // J. Am. Chem. soc. - 2009. V. 131, No. 40. - P. 14473 - 14479]. Among the structural analogues, tenuazonic acid is also known, which has cytotoxic properties [G.F. Kaufmann, R. Sartorio, S.-H. Lee [et al.]. Revisiting quorum sensing: Discovery of additional chemical and biological functions for 3-oxo-N-acylhomoserine lactones //Proc. Nat. Acad. sci. - 2005. V. 102, No. 2. - P. 309 - 314].

As a comparison standard, we took the antimicrobial drug dioxidine and the antifungal drug flucanazole [Patent No. RU 2303025 C2, patent No. RU 2690009 C1]. The objective of the invention is to obtain a new, previously undescribed benzyl ammonium salt of 4-ethoxycarbonyl-5-oxo-2-(2-oxo-2-phenylethylidene)-2,5-dihydro-1H-pyrrol-3-olate, which has an antimicrobial effect.

The claimed substance I was obtained by the interaction of 2-amino-5-(2-oxo-2-phenylethylidene)-4-oxo-1H-4,5-dihydrofuran-3-carboxylic acid ethyl ester II with benzylamine in ethanol according to the scheme:

Preparation of Compound I

10 mmol of 2-amino-5-(2-oxo-2-phenylethylidene)-4-oxo-1H-4,5-dihydrofuran-3-carboxylic acid ethyl ester was boiled in 30 ml of ethanol with the addition of 10 mmol of benzylamine for about one hour, monitoring the course of the reaction by TLC. Then the solvent was distilled off, the precipitate that formed was triturated from acetonitrile, filtered off, and recrystallized from ethanol.

Received 2.02 g of a yellow crystalline substance I (51%) with a melting point of 170-172°C. C ₂₂ H ₂₄ N ₂ O ₅ . IR spectrum, ν, cm ^-1 : 3279 (NH), 1724 (COOEt), 1694, 1646, 1609 (С=O, С=С). ¹ H NMR spectrum (DMSO-d ₆ ) δ, ppm: 1.17 t (3H, Me, J 8.0 Hz), 4.05 q (2H, CH ₂ , J 8.0 Hz), 4.90 k (2H, CH ₂ , J 6.4 Hz), 6.75 s (1H, CH), 7.26 m (5H _arom ), 7.56 m (2H _arom , J 8.5 Hz ), 7.66 m (1H _arom , J 8.5 Hz), 8.03 m (2H _arom , J 8.5 Hz) 10.01 s (1H, NH). ¹³ C NMR spectrum (DMSO-d ₆ ) δ, ppm: 14.7, 56.6, 94.6, 99.4, 126.2, 127.8, 128.3, 128.5, 128 .8, 133.0, 137.7, 138.8, 145.5, 146.5, 163.7, 166.8, 175.4, 189.7.

The claimed compound is a yellow crystalline substance, soluble in dimethylformamide, dimethyl sulfoxide, when heated - in ethanol, stable during storage.

Study of biological activity The antimicrobial properties of the claimed compound were studied on 2 collection conditionally pathogenic strains of microorganisms: Staphylococcus aureus (strain 906) and Candida albicans (PKPGY 1353/1277).

To study the antimicrobial action, the generally accepted method of two-fold serial dilutions in a liquid nutrient medium by the micromethod was used [Khabriev R.U. Guidelines for the experimental (preclinical) study of new pharmacological substances. - M.: Publishing House of Medicine, 2005. - 832 p.]. 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. In the wells of a sterile 96 well flat-bottomed microplate, two parallel rows of two-fold serial dilutions of the chemical compounds were prepared in RPB broth, Saburo. Each well contained 150 µl of the determined concentration of the test substance and 150 µl of the culture inoculum. The last rows contained nutrient medium and culture in equal volumes (control). The maximum tested concentration corresponded to 1000.0 μg/ml, the minimum - 1.0 μg/ml.

The results were evaluated using Epoch microplate Gen 5 spectrophotometer software. The last well of the row with growth retardation and OD values equal to the optical density of the control well corresponds to the minimum inhibitory concentration of the compound.

Studies have shown (see table) that compound I inhibits the growth of S. aureus bacteria at a concentration of 62.5 μg/ml, which is comparable to the reference drug, showed a fungistatic effect at a concentration of 31.2 μg/ml against yeast C. albicans.

The claimed compound - benzyl ammonium salt of 4-ethoxycarbonyl-5-oxo-2-(2-oxo-2-phenylethylidene)-2,5-dihydro-1H-pyrrol-3-olate can be used in pharmacology, medicine and veterinary medicine as potential medicines.

REFERENCES

1. Brodyck J.L. Royles. Naturally Occurring Tetramic Acids: Structure, Isolation, and Synthesis // Chem. Rev. - 1995. - V. 95, No. 6. - P. 1981-2001.

2. Colin A. Lowery, Junguk Park, Christian Gloeckner [et al.]. Defining the Mode of Action of Tetramic Acid Antibacterials Derived from Pseudomonas aeruginosa Quorum Sensing Signals // J. Am. Chem. soc. - 2009. V. 131, No. 40. -P. 14473-14479.

3. Gunnar F. Kaufmann, Rafaella Sartorio, Sang-Hyeup Lee [et al.]. Revisiting quorum sensing: Discovery of additional chemical and biological functions for 3-oxo-N-acylhomoserine lactones // Proc. Nat. Acad. sci. - 2005. V. 102, No. 2. - P. 309-314.

4. Pat. No. 2303025, Russian Federation, IPC С07С 49/16. 2,2,5,5-Tetrabromo-1,6-di-(4-methylphenyl)-1,3,4,6-hexanetraone with antimicrobial activity / N.M. Igidov, Odegova T.F., Tomilov M.V., Syropyatov B.Ya., Vakhrin M.I.; applicant and patent holder GOU VPO PGFA Roszdrav. - No. 2005123562/04; dec. 07/25/2005; publ. 07/20/2007.

5. Pat. No. 2690009, Russian Federation, IPC С07С 45/61, С07С 45/78, С07С 49/80. Method for obtaining 4,4,4-trichloro-1-(4-chlorophenyl)butan-1,3-dione with anti-candidiasis activity / N.Yu. Lisovenko, S.Yu. Balandin; applicant and patent holder federal state budgetary educational institution of higher education "Perm State National Research University". - No. 2018104421; App. 02/06/2018; publ. 05/30/2019, Bulletin No. 16.

6. Khabriev R.U. Guidelines for the experimental (preclinical) study of new pharmacological substances. - M.: Publishing House of Medicine, 2005. - 832 p.

Claims (2)

Benzylammonium salt of 4-ethoxycarbonyl-5-oxo-2-(2-oxo-2-phenylethylidene)-2,5-dihydro-1 H -pyrrol-3-olate, exhibiting antimicrobial activity,