RU2809486C2 - Drug exhibiting antihypoxic activity - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to the use of a thiotetrazole derivative

where R=methyl, Alk=N,N-diethylaminoethyl as a salt with citric acid, or R=phenethyl, Alk=N,N-diethylaminoethyl as a salt with toluenesulfonic acid, or R=phenyl, Alk=N,N-piperidinoethyl, as an antihypoxic agent.

EFFECT: invention provides for the development of compounds with high antihypoxic activity.

1 cl, 3 tbl, 3 ex, 1 dwg

Description

The invention relates to medicine, in particular to pharmacology and clinical pharmacology, and concerns the development of new pharmacological agents that increase the body's resistance to oxygen deficiency and accelerate the process of post-hypoxic recovery.

Acute hypoxia develops when exposed to various harmful factors on the body, such as low oxygen content in the inspired air, associated with an increased concentration of carbon dioxide (normobaric hypoxia with hypercapnia), exposure to substances that impede the processes of normal tissue respiration (histotoxic hypoxia) or interfere with oxygen delivery due to the deactivation of hemoglobin (hemic hypoxia). Such phenomena can occur independently (for example, when being in a hermetic volume, during accidents at chemical production, etc.), or accompany various diseases, for example, disturbances in the functioning of the cardiovascular system, intoxications of various natures, can lead to hypoxia. lung diseases, etc. In any case, hypoxia, as a damaging factor, can lead to serious disruption of the body’s functions, including its death.

The most well-known drugs that increase the body's resistance to oxygen deficiency are hypoxen, confumin and mexidol. These drugs are characterized by high dosages but low effectiveness. Hypoxen and confumin, in addition, are intended for parenteral administration. Thus, in general, the search and implementation of safe, highly effective drugs against acute hypoxia is an urgent task.

The purpose of this invention is to create compounds with high antihypoxic activity.

This goal is achieved by synthesizing chemical compounds derived from thiotetrazoles with the general formula presented in Figure 1, and in particular such compounds include:

N,N-diethyl-2-((1-methyl-1H-tetrazol-5-yl)thio)ethan-1-amine (I),

N,N-Diethyl-2-((1-phenethyl-1H-tetrazol-5-yl)thio)ethane-1-amine (II) and

1-(2-((1-Phenyl-1H-tetrazol-5-yl)thio)ethyl)piperidine (III),

where Alk=N,N-diethylaminoethyl, R=methyl (I), phenethyl (II); Alk=piperidinoethyl, R=phenyl (III).

When thiotetrazole derivatives were administered, a positive effect was observed under conditions of acute hypoxia of various origins (hypoxic with hypercapnia, acute hemic, acute histotoxic hypoxia).

Synthesis of compounds

Example 1.

N,N-Diethyl-2-((1-methyl-1H-tetrazol-5-yl)thio)ethan-1-amine (I). To a solution of 1-methyl tetrazole-5-thiol (2 g, 17 mmol) in 30 ml MeOH was added 2.96 g (17 mmol) 2-chloro-N,N-diethylethan-1-amine hydrochloride and 1.93 g (34 mmol) KOH . The solution was stirred at room temperature for 24 hours, MeOH was evaporated under vacuum. The product was purified by chromatography on silica gel CH ₃ OH-CHCl ₃ (1:10). We obtained 2.62 g of colorless oil (73%). By adding 1 equivalent of citric acid to a solution of the base N,N-Diethyl-2-((1-methyl-1H-tetrazol-5-yl)thio)ethan-1-amine in THF, a salt of the indicated compound was obtained - citrate - a colorless crystalline substance with m.p. 155-156°C ^{. 1H} NMR spectrum, δ, ppm (D ₂ O): 1.19-1.23 (6H, t), 2.64-2.81 (4H, dd), 3.18-3.24 (4H, dd), 3.45-3.58 (4H, m), 3.89 (3H, s). ¹³ C NMR spectrum, δ, ppm (D ₂ O): 8.11, 26.30, 33.70, 43.61, 47.78, 50.41, 73.78, 153.86, 174.65, 178.53. Elemental analysis, calculated: C, 44.63; N, 7.96; N, 32.53; S, 14.89. C ₈ H ₁₇ N ₅ S. Found: C, 44.48; N, 8.00; N, 32.38, S, 14.73.

Example 2

N,N-Diethyl-2-((1-phenethyl-1H-tetrazol-5-yl)thio)ethan-1-amine (II) To a solution of 1-phenethyl tetrazol-5-thiol (2 g, 9.7 mmol) in 1.65 g (9.7 mmol) of 2-chloro-N,N-diethylethan-1-amine hydrochloride and 1.93 g (34 mmol) of KOH were added to 30 ml of MeOH. The solution was stirred at room temperature for 24 hours, MeOH was evaporated under vacuum. The product was purified by chromatography on silica gel CH ₃ OH-CHCl ₃ (1:10). We obtained 2.45 g of colorless oil (83%). By adding 1 equivalent of toluenesulfonic acid to a solution of the base N,N-Diethyl-2-((1-methyl-1H-tetrazol-5-yl)thio)ethan-1-amine in THF, a salt of the indicated compound, toluenesulfonate, was obtained - a colorless crystalline substance with m.p. 81-83°C. NMR spectrum ¹ H, δ, ppm (D ₂ O): 1.13-1.18 (6H, t), 2.28 (3H, s), 3.08-3.14 (8H, m), 3.20- 2.35 (2H, m), 4.49-4.54 (2H, m), 6.92-6.96 (2H, m), 7.17-7.27 (5H, m), 7.56-7.59 (2H, d). ¹³ C NMR spectrum, δ, ppm (D ₂ O): 8.13, 20.44, 26.02, 34.68, 47.76, 49.17, 50.20, 125.30, 127.21, 128.82, 129.09, 129.39, 135.66, 1 36.69, 142.40, 154.18 . Elemental analysis, calculated: C, 58.98; N, 7.59; N, 22.93; S, 10.50. C ₁₅ H ₂₃ N ₅ S. Found: C, 59.12; N, 7.44; N, 22.88; S, 10.41.

Example 3.

1-(2-((1-Phenyl-1H-tetrazol-5-yl)thio)ethyl)piperidine (III). To a solution of 1-phenyltetrazole-5-thiol (2 g, 11.2 mmol) in 30 ml of MeOH was added 3.04 g (11.2 mmol) of 1-(2-bromoethyl)piperidine hydrobromide and 1.93 g (34 mmol) of KOH. The solution was stirred at room temperature for 24 hours, MeOH was evaporated under vacuum. The product was purified by chromatography on silica gel CH ₃ OH-CHCl ₃ (1:10). The product was obtained as a free base with m.p. 65-68°C ^{. 1H} NMR spectrum, δ, ppm (CDCl ₃ ): 1.34-1.54 (6H, m), 2.35-2.38 (4H, t), 2.64-2.67 (2H, t), 3.49 -3.352 (2H, t), 7.65-7.70 (5H, m). ¹³ C NMR spectrum, δ, ppm (CDCl ₃ ): 24.28, 25.78, 31.46, 54.03, 57.21, 125.07, 130.48, 131.06, 133.62, 155.27. Elemental analysis, calculated: C, 58.98; N, 7.59; N, 22.93; S, 10.50. Found: C, 59.12; N, 7.44; N, 22.88; S, 10.41.

Study of the biological activity of thiotetrazole derivatives

Biological activity studies were carried out in accordance with Article 11 of the Declaration of Helsinki of the World Medical Association (1964), International Recommendations for Biomedical Research Using Animals (1985) and the Rules of Laboratory Practice in the Russian Federation (Order of the Ministry of Health of the Russian Federation No. 267 dated June 19. 2003).

The antihypoxic properties of new thiotetrazole derivatives were studied in models of acute hypoxia with hypercapnia, acute hemic hypoxia and acute histotoxic hypoxia. The antihypoxic effect was assessed over the lifespan of mice in minutes. Hypoxia with hypercapnia was modeled by placing mice individually in hermetically sealed vessels with a volume of 200 cm ³ . The antihypoxic effect of chemical compounds was assessed by the life expectancy of experimental animals in an environment with a high carbon dioxide content. Hemic hypoxia was reproduced by a single injection of sodium nitrite at a dose of 230 mg/kg subcutaneously. Histotoxic hypoxia was induced by intraperitoneal injection of 0.2% sodium nitroprusside solution at a dose of 20 mg/kg into mice. When modeling hemic and histotoxic hypoxia, sodium nitrite and sodium nitroprusside were diluted in isotonic sodium chloride solution on the day of the experiment and administered to animals in a volume of 10 ml/kg.

45 minutes before modeling hypoxia, mice were intraperitoneally injected with new synthetic thiotetrazole derivatives: compounds I, II and III in doses lower than 1/10 LD ₅₀ : 25 and 50 mg/kg. Compounds I, II and III were dissolved in distilled water and/or 0.05% Tween-80 solution. The antihypoxic activity of the compounds was compared with a control group that received an isotonic sodium chloride solution in an equivalent volume, and a group that received the reference drug Mexidol, administered intraperitoneally at a dose of 200 mg/kg [Lukyanova L.D. Modern approaches to the search for antihypoxic agents // Current problems of pharmacology and the search for new drugs. T. 10. Proceedings of the conference dedicated to the 15th anniversary of the Research Institute of Pharmacology. -Tomsk, 1999. - P.59-67]. The effectiveness of the compounds was judged by the life expectancy of animals, estimated in minutes [Guide to conducting preclinical studies of drugs. Part one / ed. A.N. Mironov. - M.: Grif i K, 2012].

The statistical significance of differences was assessed using the Graph Pad Prism 6 software package. Indicators in the control and experimental groups were compared using one-way analysis of variance ANOVA and Student's t-test. Differences were considered statistically significant at p<0.05. The arithmetic mean and standard error of the mean were used to present the obtained data.

The studies were carried out on 220 outbred male mice weighing 23-27 g, obtained from the Rappolovo nursery (Leningrad region, Russia). The animals were kept under standard conditions (air temperature 21-23°C, 12-hour day/night cycle) with free access to food and water.

Research results

It has been established that the magnitude and severity of the antihypoxic effect depend on the compound, dose and model of hypoxia.

In the test of acute hypoxia with hypercapnia, the death of mice in the control group was observed after 18.8±0.5 minutes. Mexidol at a dose of 200 mg/kg increased the life expectancy of animals by 31% compared to the control group. Compound I at doses of 25 and 50 mg/kg increased this indicator by only 17 and 19%, respectively. When II was administered at a dose of 25 mg/kg, a slight antihypoxic effect was observed, and when the drug was administered at a dose of 50 mg/kg, the life expectancy of mice increased by 52% compared to the control group. Compound III had antihypoxic activity, and its activity was similar to the drug Mexidol (Table 1).

Under conditions of acute hemic hypoxia, the life expectancy of mice in the control group was 22.5±1.5 minutes. The reference drug Mexidol and compound I (25 and 50 mg/kg) slightly increased this indicator, and compound III turned out to be inactive. Compound II exhibited a pronounced antihypoxic effect under conditions of acute hemic hypoxia at doses of 25 and 50 mg/kg, significantly increasing the life expectancy of mice by 64 and 44%, respectively (Table 2).

With histotoxic hypoxia, the death of mice occurred after 10.0±0.8 minutes. Mexidol increased this indicator by 43%. With the prophylactic administration of compounds I and II, both at a dose of 25 mg/kg and 50 mg, life expectancy significantly increased by 52.45 and 50.52%, respectively (Table 3).

Claims (3)

Application of thiotetrazole derivative formula where R=methyl, Alk=N,N-diethylaminoethyl as a salt with citric acid, or R=phenethyl, Alk=N,N-diethylaminoethyl as a salt with toluenesulfonic acid, or R=phenyl, Alk=N,N-piperidinoethyl, as an antihypoxic agent.

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