RU2192271C1 - Method of cardiotropic agent preparing - Google Patents

Abstract

FIELD: medicine, cardiology, phytotherapy, pharmacy. SUBSTANCE: method involves preparing liquid extract from common fenugreek seeds (Trigonella foenum-graecum) by extraction with 95% ethyl alcohol in the ratio raw : extractant = 1:1 at 18-20 C for 144 h in 6 diffusers. Invention ensures to obtain product devoicing the hepatotoxic effect. EFFECT: improved method of preparing, absence of adverse effect. 3 tbl, 6 ex

Description

 The invention relates to medicine and relates to methods for producing preparations from plant materials, which can be used to treat cardiovascular diseases accompanied by chronic heart failure.

 Known means that improve the functional state of the heart during its decompensation - lantoside, celanide, digoxin, cordigit and others [Radar-Pharmacist / Ed. Krylova Yu.F. - M., 2001. - 1584 s .; Sokolov S.Ya., Zamotaev I.P. Handbook of medicinal plants. M.: Medicine, 1985. - S. 81-83; Mashkovsky M.D. Medicines In 2 volumes - M .: Medicine, 1988. - T. 1. - S. 395.].

 The closest in nature of origin (prototype) is the drug adoniside (Novogalenovy preparation from spring adonis herb), which has cardiotropic activity. Adoniside by the nature of the action takes an intermediate place between strophanthin - K and digitalis preparations. It has a positive inotropic effect on the heart (an increase in contractility function), a negative chronotropic effect (a decrease in heart rate) and a negative bathmotropic effect (a decrease in the excitability of the heart muscle). A known method of obtaining a cardiotropic drug adoniside, which consists in the extraction of plant raw materials of adonis spring mixture of chloroform and 95% ethyl alcohol in a ratio of 95: 5, followed by purification from ballast substances [Technology of dosage forms: 2 tons / Ed. Ivanova L.A. - M .: Medicine, 1991. - T.1. - C. 430-431].

 Adoniside, digitalis preparations are glycosides and therefore their cumulative and hepatotoxic effects are a significant drawback. In this regard, prolonged use of such drugs is undesirable and it is recommended that they be alternated with other drugs in order to avoid side effects.

 However, the list of drugs with a cardiotropic effect is limited [Mashkovsky MD, 1988; Radar Pharmacist. 2001] due to the lack of an appropriate plant base. Therefore, the expansion of the arsenal of herbal preparations, characterized by cardiotropic properties, is an urgent task.

 The aim of the invention is to develop a method for producing a cardiotropic agent from fenugreek, characterized by the absence of hepatoxicity.

The task is achieved in that in order to obtain the target product, the feedstock — the seeds of hay fenugreek (Trigo-nella foenum graecum L.), are treated with 95% ethanol at a temperature of 18-20 ^o C, with a ratio of raw material: extractant 1: 1 for 144 hours in six diffusers.

 There is no data in the literature on the possibility of using hay fenugreek extract as a cardiotropic agent. The proposed cardiotropic agent can be used in medical practice for the treatment of cardiovascular diseases accompanied by circulatory disorders. This property does not explicitly follow for a person skilled in the art. Thus, the proposed solution meets the criteria of the invention - "novelty", "inventive step", "industrially applicable".

 The use of 95% ethyl alcohol as an extractant is due to the most complete extraction of active substances. The temperature regime, the ratio of raw materials: extractant, the number of diffusers, exposure time are optimal and are selected experimentally.

A cardiotropic agent is obtained as follows: crushed hay fenugreek seeds (Trigonella foenum graecum L.) and extractant, 95% ethyl alcohol are divided into six equal parts. The first portion of the raw material is extracted with a pure extractant, and each subsequent (second, third, fourth, fifth, sixth) is extracted with an extract obtained by extraction of the previous portion of the raw material. The ratio of raw materials: extractant is 1: 1, taking into account the absorption of raw materials. The duration of the process is 144 hours. The obtained extract (liquid extract) is filtered through 4 layers of gauze, standing at a temperature not exceeding 10 ^o C for at least 2 days. The extract yield for active substances is 77.2%. The content of steroid compounds in the preparation, in terms of diosgenin, is 2.07 ± 0.001%.

 An example of a specific implementation of the method.

 Example.

 The extract was prepared in six percolators. In each of the six percolators, crushed hay fenugreek seeds weighing 20.0 kg were placed. The volume of extractant introduced into the battery of percolators at each stage of extraction is 40.2 liters. The volume of each extract, selected from the sixth, head percolator, is 20 liters.

 1 day. Percolators 1 and 2 were introduced into the work. At 8 o’clock in the morning, 40.2 l of extractant was poured into percolator 1 and left to infuse until 16 hours.

 At 16 o’clock, the percolator 1 valve was opened and the extraction was poured into the collector 1. 40.2 L of extractant was introduced into the percolator 1 and the liquid phase was continued to drain until it was completely accumulated in the 20 collector 1. After that, the percolator 1 cock was closed, and the extract from the collection 1 was poured into the percolator 2. Percolators 1 and 2 were left to infuse until 8 am the next day (about 16 hours).

 2 day. Percolators 3 and 4 were introduced into the work. At 8 a.m. the cranes of percolators 1 and 2 were opened and the extracts were drained. 40.2 l of extractant was introduced into percolator 1 and the extraction was continued to drain until it was completely accumulated in collector 1 with a volume of 20 l. After that, the percolator valve 1 was closed. Extract from collector 1 was transferred to percolator 2 and 20 liter extract was collected in collector 2, then the percolator 2 valve was closed. Extract from collector 2 was transferred to percolator 3 and left to infuse until 16 hours.

 At 16 o'clock, cranes of percolators 1-3 were opened and the extracts were drained. 40.2 l of extractant was introduced into percolator 1 and the extraction was continued to drain until it was accumulated in collector 1 with a volume of 20 l. After that, the percolator valve 1 was closed. The extract from the collection 1 was transferred to the percolator 2. The drain of the extraction from the collection 2 of 20 l was transferred to the percolator 3, and the drain of the extraction from the collection 3 of 20 l was transferred to the percolator 4. Percolators 1-4 were left to infuse until 8 am the next day.

 3 day. Percolators 5 and 6 were introduced into the work. At 8 a.m., the cranes of percolators 1 and 2 were opened and the extraction was drained. 40.2 L of extractant was introduced into percolator 1. Draining the extract from the collector 1 with a volume of 20 l was transferred to the percolator 2. Draining the extract from the collector 2 with a volume of 20 l was transferred to the percolator 3, draining the extract from the collector 3 with a volume of 20 l was transferred to the percolator 4. Draining the extract from the collector 3 with a volume of 20 l was transferred to the percolator 5. Percolators 1-5 were left to infuse until 16 hours.

 At 16 o'clock cranes of percolators 1-5 are opened and discharge from the first portion of extraction was performed. 40.2 L of extractant was introduced into percolator 1. Draining the extract from collector 1 with a volume of 20 ml was transferred to percolator 2, draining the extract from collector 2 with a volume of 20 ml was transferred to percolator 3, draining the extract from collector 3 with a volume of 20 ml was transferred to percolator 4. Draining the extract from collector 3 with a volume of 20 ml was transferred to percolator 5. Drain the extract from the collection 5 with a volume of 20 ml was transferred to the percolator 6. Percolators 1-6 were left to infuse until 8 o’clock the next day.

 4 day. Take 1 and 2 servings of the extract, take out the percolators 1 and 2. At 8 a.m. the cranes of percolators 1-6 were opened and the extracts were drained. The volume of each extract should be 20 liters. Extract 13 of collection 6 was transferred to a sump. Percolator 1 was taken out of operation. The depleted feed from percolator 1 was unloaded and transferred to a flask for distilling off the alcohol. Extracts from collections 1-5, each 20 ml in volume, were transferred to the next extraction stage, in percolators 2-6, and left to infuse until 16 hours.

 At 16 o'clock the cranes of percolators 2-6 were opened and the extracts were drained. Extract from collection 6 was transferred to a sump. Percolator 2 was taken out of operation. The depleted feed from percolator 2 was unloaded and transferred to a flask to distill off the alcohol. Extracts from collectors 2-5, each 20 ml in volume, were transferred to the next extraction stage in percolators 3-6 and left to infuse until 8 am the next day.

 5 day. 3 and 4 portions of the extract were removed, percolators 3 and 4 were taken out of operation. At 8 o’clock in the morning, cranes of percolators 3-6 were opened and the extracts were drained. The volume of each extract should be 20 ml. Extract 13 of collection 6 was transferred to a sump. Percolator 3 was unloaded and the depleted feed was transferred to a flask for distilling off the alcohol. Extracts from 3-5 collectors, each 20 ml in volume, were transferred to the next stage of extraction percolators 4-6 and left to infuse until 16 hours.

 At 16 o'clock, cranes of percolators 4-6 were opened and the extracts were drained. Extract from collection 6 was transferred to a sump. Percolator 4 was taken out of operation. The depleted feed from percolator 4 was unloaded and transferred to a flask to distill off the alcohol. Extracts from collectors 4-5, each 20 ml in volume, were transferred to the next extraction stage in percolators 5-6 and left to infuse until 8 o’clock the next day.

 6 day. 5 and 6 portions of the extract were removed, percolators 5 and 6 were taken out of operation. At 8 o’clock in the morning, cranes of percolators 5-6 were opened and the extracts were drained. The volume of each extract should be 20 ml. Extract from collection 6 was transferred to a sump. Percolator 5 was taken out of operation. The depleted feed from percolator 5 was unloaded and transferred to a flask for distilling off alcohol. Extracts from collectors 5, with a volume of 20 ml, were transferred to percolator 6 and left to infuse until 16 hours.

At 16 o'clock percolator cranes 6 were opened and extraction was drained. Extract from collection 6, with a volume of 20 ml, was transferred to a sump. Percolator 6 was taken out of operation. The depleted feed from percolator 6 was unloaded and transferred to a flask to distill off the alcohol. All six extracts were combined, mixed and defended at a temperature not exceeding 10 ^o C for 2 days, after which they were filtered off and quality assessment was carried out.

 To establish the cardiotropic properties of fenugreek extract, 2 series of experiments were carried out. The cardiotropic effect was studied by indicators of chronotropic and inotropic effect.

 The chronotropic effect was evaluated by the number of heart contractions per minute. The experiment was carried out on white outbred rats of both sexes, kept in the general vivarium mode, weighing 200-220 g using an EC1T-0.3M cardiograph. The extract solution was injected in a volume of 1.0 ml into the stomach through a tube. The cardiogram was taken before administration and 2 hours after administration of the substance. To exclude the effect of high alcohol concentrations (95%) on the gastric mucosa and central nervous system, the extract was evaporated to 1/3 of the volume, and then the volume was adjusted to the initial saline. In the control, 30% ethanol was added.

 The inotropic effect was evaluated by an electrocardiogram (ECG) according to the degree of shortening of the Q-T interval and a decrease in the voltage of the T wave. In the group of animals receiving the extract at a dose of 20 mg / kg, a shortening of the Q-T interval and a decrease in the voltage of the T wave were observed, which indicates an increase in the positive inotropic effect. Since a quantitative assessment of changes in the nature of the cardiogram is difficult, the inotropic effect was additionally evaluated using a myograph. The experiments were carried out on an isolated frog heart, on which a 1% solution of hay fenugreek extract was applied.

 Statistical processing of the results of all series of experiments was carried out using the method of variation statistics using the student coefficient. The research results are presented in examples 1-5.

 Examples of cardiotropic activity of the drug.

 Example 1

 The introduction of experimental groups of animals (rats) extract of fenugreek hay at a dose of 20 mg / kg led to a decrease in heart rate (HR) by 32.8% compared with the control (table 1).

 Example 2

 The introduction of experimental groups of animals (rats) of adoniside at a dose of 20 mg / kg led to a decrease in heart rate (HR) by 32.1% compared with the control (table 1).

 Example 3

 Application of a 1% solution of hay fenugreek extract on an isolated heart of experimental groups of animals (frogs) led to an increase in heart rate (CVS) by 18.91% compared with the control (table 2).

 Example 4

 Application of a 1% solution of adoniside on an isolated heart of experimental groups of animals (frogs) resulted in an increase in heart rate (CVS) of 17.35% compared with the control (table 2).

 Thus, hay fenugreek extract, prepared using 95% ethanol, has a pronounced cardiotropic effect and is not inferior to the prototype (adoniside) in its ability to reduce heart rate and increase their strength.

 In parallel with the study of the cardiotropic effect of hay fenugreek extract, we studied its acute toxicity. The results of the study are presented in example 5.

 Example 5

 The experiments were carried out on white outbred mice weighing 15-20 g. The extract was concentrated and then injected into the mice in the form of an aqueous solution inside through a probe. Doses of 1000 were studied; 1500; 2000; 2500; 3000 mg / kg. Observation of the animals was carried out for 7 days. For each dose, 6 animals were taken. In group I, animals treated with the test substance at a dose of 1000 mg / kg, there were no dead animals during the observation period. In group II, 1 animal died (dose 1500 mg / kg). In group III (dose 2000 mg / kg) 50% of the animals died (out of 6 animals - 3), in group IV (dose 2500 mg / kg) - out of 6 animals -4. In group V (dose 3000 mg / kg), all animals died. Thus, based on calculations by Kerber, acute toxicity is 2083 mg / kg. The calculations are presented in table 3.

 Thus, hay fenugreek extract belongs to the 3rd class of harmful substances according to hazard level 1 (moderately hazardous substances) [Treatment of acute poisoning / Ed. M.L. Tarakhovsky - Kiev: Health, 1982. - P. 7] or to the class of "low toxicity substances" by combined tabulation of toxicity classes (according to Hodge and Sterner) [Methods for determination of toxicity and hazard of chemicals / Ed. I.V. Sanotsky. - M .: Medicine, 1970. - P.16].

Claims (1)

A method of obtaining a liquid extract from plant material with a cardiotropic effect, characterized in that the hay fenugreek seeds are extracted with 95% ethyl alcohol at a ratio of raw materials: extractant = 1: 1 for 144 hours at 18-20 ^{° C.} in six diffusers.

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