EA002379B1 - Remedy having anti-ischemic, antihypoxic and hypotensive activities - Google Patents

Abstract

1. The remedy having antihypoxic activity, comprising N-carbamoylmethyl-4-phenyl-2-pyrrolidone, characterized in that it additionally comprises dibasic- or tribasic carboxylic acid. 2. The remedy according Claim 1, characterized in that it comprises oxalic acid or malic acid or siccine acid as a dibasic acid. 3. The remedy according Claim 1, characterized in that it comprises citric acid as a tribasic acid. 4. The remedy according Claims 1,2,3, characterized in that molar ratio N-carbamoylmethyl-4-phenyl-2-pyrrolidone: carboxylic acid is 1(0,5/2,0).

Description

The invention relates to the pharmaceutical industry and relates to agents affecting the cardiovascular system.

Various biologically active compounds and their compositions are known that affect the cardiovascular system. The agent [1] is described, which is 3-methyl-5 [2- (3-ter. Butylamino-2-hydroxypropoxy) phenoxymethyl] -1,2,4-oxadiazole, which exhibits antihypertensive, antiarrhythmic, coronary expansion, anti-ischemic and anti-glaucoma activity. Its anti-ischemic activity is not great, antihypoxic activity is not described.

One of the widely used agents with anti-ischemic activity is obzidan (anaprilin) [2]. However, its use is limited in patients with obstructive pulmonary diseases due to the bronchospasm effect and in patients with obliterating diseases of the peripheral vessels due to the presence of a negative inotropic effect.

The most famous antihypoxic agent is piracetam [3]. It has a positive effect on metabolic processes and blood circulation. There is data [4] also on its positive effect in patients with coronary heart disease, but only in old and old age.

Succinic acid and its salts are known to have adaptogenic ability and have antihypoxic, anti-stress and neurotropic effects, normalize energy metabolism. The cardiotropic and anti-ischemic action of succinic acid is also known [5]. The hypotensive effect of succinic acid is not described.

Closest to the proposed invention is I-carbamoylmethyl-4-phenyl2-pyrrolidone, exhibiting antihypertensive [6] and antihypoxic activity [7, 8]. Its anti-ischemic activity is not described.

The objective of the invention is to create an effective low-toxic drug with anti-ischemic, antihypertensive and antihypoxic activity, expanding the arsenal of cardiovascular agents.

The problem is realized by the proposed tool, including Ν-carbamoylmethyl-4-phenyl-2-pyrrolidone and dibasic or tribasic carboxylic acid. As the dibasic acid, either oxalic, or malic, or succinic acid is used; as the tribasic acid, citric. Moreover, the ratio of No. carbamoylmethyl-4-phenyl-2-pyrrolidone (CFP): carboxylic acid is 1: (0.5-2) mol.

The invention is illustrated by the following examples.

Example 1

In a two-necked flask equipped with a stirrer and a reflux condenser, 60-100 ml of alcohol (ethyl, methyl, isopropyl) is placed, then the corresponding amounts of carbamoylmethyl-4-phenyl-2-pyrrolidone (CPP) and carboxylic acid are added. The mixture is heated to boiling, then cooled to room temperature and left under stirring for 1-1.5 hours. The precipitated crystalline precipitate is separated and dried.

The number of components are given in table. one.

Example 2

The ball mill is charged with the appropriate amounts of CPP and carboxylic acid. The mixture is thoroughly stirred for 0.5-1 h, then the resulting product is discharged.

The amounts of the components are the same as in example 1.

Table 1. The ratio of No. carbamoylmethyl-4-phenyl-2pyrrolidone (CPP) and carboxylic acids

CFP, g (mol) Oxalic acid, g (mol) Succinic acid, g (mol) Malic acid, g (mol) Citric acid, g (mol) 10.9 (0.05) 9.0 (0.1) 11.8 (0.1) 13.4 (0.1) 21.0 (0.1) 21.8 (0.1) 9.0 (0.1) 11.8 (0.1) 13.4 (0.1) 21.0 (0.1) 32.7 (0.15) 9.0 (0.1) 11.8 (0.1) 13.4 (0.1) 21.0 (0.1) 43.6 (0.2) 9.0 (0.1) 11.8 (0.1) 13.4 (0.1) 21.0 (0.1)

The output in all examples is quantitative.

The ratio of CFP: carboxylic acid is 1 mol: (0.5-2).

All of the following compositions were studied in experiments on animals in models for anti-ischemic, antihypertensive and antihypoxic activity.

No. carbamoylmethyl-4-phenyl-2-pyrrolidone + oxalic acid (KPP-SH) No. carbamoylmethyl-4-phenyl-2-pyrrolidone + succinic acid (KPP-YAN) No. kabamoylmethyl-4-phenyl-2-pyrrolidone + malic acid ( KFP-YAB) No. carbamoylmethyl-4-phenyl-2-pyrrolidone + citric acid (KFP-L)

Anti-ischemic activity of CFP and proposed agents

The anti-ischemic effect was studied in a model of short-term ischemia caused by occlusion of the descending branch of the left coronary artery (VDVL) at the border of the upper and middle third in cats and rats (for CFP). The effect of the drugs was assessed by their effect on the dynamics of the total increase in the 8T segment of the epicardial electrogram recorded from three points of the heart surface located in the ischemic zone, at the border of the ischemia and intact zone, in the intact region, which reflects the functional state of the ischemic focus. Obzidan was used as a comparison drug.

CPP was injected into the peritoneum at a dose of 50 mg / kg 30 minutes before ONLVA, the myocardium of the animals was examined using the morphometric method, determining the area of the necrosis zone, ischemia zone, and total myocardial lesion zone (necrosis + ischemia) in the experimental and control groups (with the introduction of fiz. solution 10 minutes before ONLVA and obzidan at a dose of 0.5 mg / kg 30 minutes before ONVLKA). The results of the study are presented in table. 2.

Table 2. The results of studies on the effect of CFP on the area of myocardial damage in experimental rat-induced heart attack caused by ONCLA

Group The area of the myocardium, mm ² Area of ischemia zone, _mm ² The ratio of ischemia / myocardium The area of the necrosis zone, _mm ² The ratio of the area of necrosis / myocardium The area of the affected area, _mm ² The ratio of necrosis / ischemia Fiz. rr 485.97 ± 4.87 233.09 ± 11.34 0.479 ± 0.022 36.00 ± 5.84 0.074 ± 0.012 269.09 ± 9.07 0.165 ± 0.036 Obzidan 481.13 ± 8.12 175.21 ± 4.01 * 0.365 ± 0.009 * 28.48 ± 4.08 0.059 ± 0.009 203.68 ± 6.08 * 0.163 ± 0.024 CFP 492.55 ± 4.85 163.33 ± 7.84 * 0.332 ± 0.017 * 19.21 ± 6.35 0.039 ± 0.013 182.53 ± 12.16 * 0.113 ± 0.036

* - The differences are significant compared with the group receiving saline.

These results show that KPF has a pronounced anti-ischemic activity that exceeds that of obzidan.

The test substances at a dose of 50 mg / kg and the reference drug - obzidan at a dose of 0.25 mg / kg, were administered 5 min before ONVLK. In all groups of the studied substances, a significant decrease in the total increase in the 8T segment was revealed. The greatest effect was observed with the introduction of KFP-U, the introduction of which caused an almost 50% decrease in the total increase in the 8T segment by the fifth minute after ONVLK. This effect exceeded that of obzidan. The results are presented in table. 3.

Table 3. The effect of the proposed funds on the total 8T segment elevation in cats after ONVLK

Drug name Originally Time after ONLK, min Time after removal of ONVLK, min 0.5 one 3 5 0.5 one 3 5 Fiz. rr 1.7 ± 0.4 3.5 ± 0.8 3.6 ± 1.1 4.8 ± 0.8 4.9 ± 1 4.1 ± 1 3.6 ± 0.6 2.5 ± 0.9 2.4 ± 1 KFP-SH 1.7 ± 0.6 1.9 ± 0.4 * (- 45.7) 1.9 ± 0.6 (-47.2) * 2.5 ± 0.6 (-47.9) * 2.4 ± 0.6 (-51) * 1.9 ± 0.5 (-53.7) * 1.7 ± 0.8 (-52.8) * 1.4 ± 0.5 (-44) * 1.2 ± 0.5 (-fifty)* KFP-L 1.7 ± 1.3 2.0 ± 1.2 (-42.9) * 2.0 ± 1.3 (-44.4) * 2.5 ± 1.2 (-47.9) * 2.6 ± 1.1 (-46.9) * 2.6 ± 0.8 (-36.6) * 2.7 ± 1.2 (-25.0) * 1.9 ± 1.1 (-24.0) * 1.5 ± 1.6 (-37.5) * KFP-YAB 1.7 ± 0.3 2.3 ± 0.3 2.4 ± 0.5 2.5 ± 0.8 2.6 ± 0.0 2.5 ± 0.6 1.9 ± 0.3 1.8 ± 0.4 1.6 ± 0.3 * (- 34.3) (-33.3) * (-47.9) * (-46.9) (-39.0) * (-47.2) * (-28.0) * (-33.3) * KFP-YAN 1.7 + 0.9 2.2 + 1.3 2.7 + 0.6 2.8 + 1.8 3.3 + 1.5 2.3 + 1.2 1.9 + 0.8 1.3 + 0.6 1.3 + 0.6 (-42.9) (-25.0) (-41.7) (-36.7) (-43.9) (-47.2) (-48.0) (-45.8) Obzidan 1.7 ± 1.4 2.8 ± 1.6 2.9 ± 1.7 5.5 ± 2.1 5.6 ± 2.2 5.9 ± 2.5 4.7 ± 1.9 3.4 ± 2.0 3.5 ± 2.3 (-20.0) (-19.4) (+14.6) (+14.3) (+43.9) (+30.6) (+36.0) (+45.8)

* - Statistically significant (p <0.05) compared with the control group (saline solution).

Thus, it can be seen from the above data that all the proposed agents exhibit pronounced anti-ischemic activity in excess of this activity of CPP and obzidan.

Antihypertensive activity

It was previously shown [6] that CPP exhibits hypotensive activity in various types of arterial hypertension, which is confirmed by the following data.

In experiments on anesthetized and non-anesthetized cats, CPP in doses of 25-50 mg / kg lowered blood pressure (BP) by 15-20%, the maximum effect - by 60-90 minutes. At the same time, a decrease in total peripheral resistance (OPS) by 26 ± 7.1% and an increase in minute blood volume (IOC) by 14.9 ± 3.8%. The method for determining the IOC - thermal dilution according to Red1et (1954) in the modification of M.I. Gurevich (1967), D.E. Vankova and Yu.I. Qibina (1973).

All the proposed means in experiments on cats showed a clear antihypertensive effect, the most active was the composition of KPP-YAN. The drug KFP-YAN in experiments on cats gives a clear hypotensive effect (at a dose of 50 mg / kg, an intravenous decrease in blood pressure (BP) by 18-20%, maximum effect by 60-90 minutes, a decrease in total peripheral resistance by 38%) and reduces blood pressure (by 28.2%) and heart rate (by 15%) when injected into the ventricles of the brain of rats at a dose of 10-500 μg, which indicates its effect on the central mechanisms of blood circulation regulation. The realization of this effect probably occurs with the participation of GABA-ergic (suppressing the hypotensive effect of CPP-YAN with biculin and picrotoxin) and noradrenergic (suppressing the effect of phentolamine, and _2- adrenergic blocker yohimbine and preliminary administration of reserpine) brain systems.

In the renal (25 cats), adrenal regenerative (25 cats) and neurogenic (25 cats) models of arterial hypertension (AH), a moderate but prolonged decrease in blood pressure was observed due to the administration of CPP-YAN (50 mg / kg, iv) decrease in total peripheral resistance (OPS) while increasing cardiac output. Discontinuation of treatment did not cause withdrawal syndrome (Table 4).

Table 4. The antihypertensive effect of CPP-YAN with various models of arterial hypertension (p <0.05)

Type of AG BP reduction (%) Decrease in OPS (%) during treatment Not too late. after the introduction of the teacher Across 2 weeks treatment Across 1 week by- after cancellation Adrenal regenerative 23.3 25.3 25,2 32,5 Neurogenic 21.1 28.1 23,2 39,4 Renal 1.3 28.3 28,2 39,4

Compositions KFP-L, KFP-Shch and KFP-YaB also showed a distinct antihypertensive effect, comparable to the effect of KFP-YAN and exceeding that of KFP.

Antihypoxic activity

The antihypoxic activity of CPP was studied on a model of normobaric hypoxic hypoxia [8] in male mice weighing 20-30 g. The protective effect was evaluated by the life span of animals, CPP increased the life span of mice in a pressure chamber, i.e. under normobaric hypoxic hypoxia, it has an antihypoxic effect, unlike piracetam, which does not exhibit such properties.

The antihypoxic activity of the proposed drugs was studied under conditions of acute hypoxic hypoxia in order to more deeply and in detail evaluate the antihypoxic effect. All of the proposed funds showed a pronounced antihypoxic activity, higher than that of CPP. The most active was the composition of KPP-YAN.

In conditions of acute hypoxic hypoxia, KPF-YAN, administered intraperitoneally to rats, inhibits a decrease in the activity of the enzymes of the glutathione antioxidant system (glutathione reductase and glutathione peroxidase by 19.3 and 30.5%, respectively), 5'-nucleotidase (by 9.4%), Ba ⁺ , K ⁺ - ATPase (by 24.9%) and has a depriminating effect on the activity of lysosomal enzymes (cathepsin Ό and β-glucosidase by 25 and 32.8%, respectively, compared with the control group) and the intensity of peroxide processes lipid oxidation (LPO) in rat myocardium; under conditions of ischemic damage to the myocardium and brain (carotid ligation) KFP-YAN (50 mg / kg, iv) caused a significant decrease in diene conjugates, free fatty acids and malondialdehyde in the myocardium and brain compared with the control group, table . 5 and 6.

Table 5. The effect of CFP-YAN on the parameters of lipid peroxidation and the content of energy substrates in experimental myocardial and cerebral ischemia

Experimental myocardial infarction card ATP ADP The sum of the nucleotides AMF Myocardium Without treatment 433.3% 431.5% 423.9% 4116.7% KFP-YAN 440.0% 421.6% 426.8% 434.7% Brain Without treatment 437.1% 426.5% 426.8% nd KFP-YAN 430.0% 438.2% 431.1% nd Cerebral ischemia ATP ADP The sum of the nucleotides AMF Brain Without treatment 422.4% 422.2% 428.2% 483.0% KFP-YAN 485.9% nd 451.9% 431.8%

Note: 4 - increase, 4 - decrease, nd - data are not reliable;

(p <0.05 compared with data before ischemia).

Table 6. The decrease (%) in the content of lipid peroxidation products on the background of the administration of KFP-YAN 50 mg / kg / peritoneum in experimental rat myocardial and cerebral ischemia

Products FLOOR Diene conjugates, µmol / l Free fatty acids, µmol / L Malondialdehyde, mmol / L Myocardium Head brain Myocardial ischemia 18.8 23.9 23.9 47.6 Cerebral ischemia 33.9 29.3 29.3 37.5

In a study of the myocardial oxygen consumption by the Warburg manometric method, it was shown that in rats under conditions of acute hypoxic hypoxia, the prophylactic administration of CPP-J in / peritoneum leads to an increase in oxygen consumption by the myocardium, liver and cerebral hemispheres. Consequently,

KPP-YAN prevents the formation of "oxygen debt" in the studied organs, contributing to the extraction of animal tissues from the blood of the required amount of oxygen.

Table 7. The effect of CFP-YAN (50 mg / kg, intraperitoneally) and cordarone (10 mg / kg, intraperitoneally) on oxygen consumption by the myocardium, liver and cerebral hemispheres

Group Oxygen consumption (μl / h / mg dry matter) Myocardium Liver Brain The initial state 4.8 ± 0.2 4.3 ± 0.1 6.0 ± 0.1 KFP-YAN 4.8 ± 0.1 4.4 ± 0.2 6.1 ± 0.1 Cordaron 4.7 ± 0.2 4.3 ± 0.2 5.9 ± 0.2 Hypoxic hypoxia (GG) 6.0 ± 0.1 5.3 ± 0.2 7.2 ± 0.1 KFP-YAN + GG 5.0 ± 0.2 * 4,5 ± 0,1 * 6.3 ± 0.2 * Cordaron + GG 5.0 ± 0.1 * 4.4 ± 0.2 * 5.8 ± 0.2 *

* - Data are reliable regarding hypoxic hypoxia (p <0.05).

CFP-YAN, previously introduced in / peritoneally, prevents the development in rat myocardium of changes associated with the damaging effect of acute hypoxic hypoxia on the ultrastructural organization of the hematoparenchymal barrier and mitochondrial apparatus (data from pathamorphological studies). The remaining proposed means had a similar effect.

The results of the study of acute toxicity showed that for all the proposed means, LI 50 when administered intraperitoneally to mice is 1050-1340 mg / kg (LI 50 CFP 1030 mg / kg).

The proposed funds are low toxic, they can be used in any suitable dosage forms containing an effective amount of the proposed funds and pharmacologically acceptable carriers and / or excipients.

Thus, the proposed tools have anti-ischemic, antihypertensive and antihypoxic activity exceeding the similar activity of Y-carbamoylmethyl-4-phenyl-2-pyrrolidone (CPP) and a number of known drugs. In addition, the presence of anti-ischemic activity in combination with antihypertensive and antihypoxic drugs makes these drugs promising for the treatment of cardiovascular diseases.

Literature

1. RF patent No. 1827067, A 61K 31/41, BI No. 19, 1994

2. M.D. Mashkovsky. Medicines Kharkov: Torsing, 1997, v. 1, p. 262-264.

3. In the same place. from. 109-111.

4. L.A. Leshchinsky, L.G. Pimenov, V.S. Fedorov. Therapeutic use of piracetam in patients with myocardial infarction // Cardiology, 1987, No. 2, p. 46-49.

5. Succinic acid in medicine, food industry, agriculture. Collection of scientific articles. Pushchino, 1997

6. A.S. USSR No. 797219, BI No. 21, 1995, A 61 K 31/41, S07 D 202/26.

7. Yu.G. Bobkov, I.S. Morozov, O.M. Glozman et al. Pharmacological characteristics of the new phenyl analogue of piracetam-4 phenylpiracetam. // Bull.exper.biology and medicine. 1983, KhSU, p. 50-53.

8. RF patent No. 2050851. A 61 K 31/40, BI No. 36.1995,

Claims (4)

CLAIM 1. The tool has anti-ischemic, antihypertensive and antihypoxic activity, including Y-carbamoylmethyl-4-phenyl-2-pyrrolidone, characterized in that it additionally contains dibasic or tribasic carboxylic acid. 2. The tool according to claim 1, characterized in that as the dibasic acid, it contains oxalic, or malic, or succinic acid. 3. The tool according to claim 1, characterized in that as a tribasic acid, it contains citric acid. 4. The tool according to claims 1 to 3, characterized in that the ratio of Y-carbamoylmethyl-4-phenyl-2pyrrolidone: carboxylic acid is 1: (0.5-2.0) mol.