RU2328302C2 - Medicinal agent of antihypoxic action - Google Patents

Abstract

FIELD: medicine; pharmacology.

SUBSTANCE: medicinal agent of antihypoxic action is produced by extraction from dried and milled candle larkspur herb with ethyl alcohol at specified conditions. Medicinal agent of antihypoxic action is produced by extraction from dried and milled candle larkspur herb with chloroform and represents total alkaloids.

EFFECT: agents of evident antihypoxic activity.

3 cl, 1 tbl, 3 ex

Description

The invention relates to medicine, specifically to pharmacology, and can be used as an antihypoxic agent in the treatment of various diseases associated with impaired functions of the respiratory, cardiovascular systems, liver, kidneys, blood transport function, as well as poisoning with various poisons [1, 2].

The closest to the positive effect (basic) is α-tocopherol acetate. This drug has an antioxidant effect, participates in tissue respiration and other important processes of tissue metabolism, but it is a synthetic drug and has a number of side effects [3]. The closest means with antihypoxic action by nature of origin is: 10% tincture of the stalks of elecampane high [4]. However, at present, the issue of expanding the arsenal of antihypoxic agents from natural raw materials is extremely relevant.

The objective of the invention is the expansion of the arsenal of funds with antihypoxic effect from natural raw materials.

The set task is achieved by the fact that the antihypoxic agent contains an alcoholic extract from the aerial part (grass) of the high luminosity (Delphinium elatum. L), in particular a liquid extract from the dried and ground aerial part (grass) of the larkspur high 40% ethyl alcohol in the ratio of vegetable raw materials and extractant 1:10, and a liquid extract from the dried and ground aerial parts (grass) of the livestock with high 40% ethyl alcohol with a ratio of plant materials and extractant 1: 2, as well as a dry extract of the amount of alkaloids, highlighted high from dried and ground aerial parts (grass) of high liveliness.

It was shown for the first time that products containing alcohol extracts from dried and ground aerial parts (grass) of livestock with high 40% ethanol in the proposed ratio, as well as the amount of alkaloids, have an antihypoxic effect. The authors did not find in the literature analyzed by them information about the antihypoxic effect of alcohol extracts from the aerial part (grass) of high vivacity. The antihypoxic effect of alcoholic extracts from the aerial part (grass) of the ankle highly clearly does not follow from the prior art in this field and is not obvious to a specialist. New properties have been experimentally established and the possibility of using alcohol extracts from dried and ground grass of high larkiness as a means with antihypoxic activity in medicine.

Thus, the present invention meets the criteria of patentability "novelty", "inventive step", "industrial applicability".

Antihypoxic agents are alcohol extracts from high liveliness, containing a complex of active and related substances extracted from ground grass of livestock with a high 40% solution of ethyl alcohol, followed by settling and filtration, as well as an alkaloid extract extracted with chloroform and containing the amount alkaloids.

The funds we offer are obtained as follows.

Example 1. The extraction is carried out by maceration. The grass of high livestock collected in July is dried, ground to 3 mm, 10 g of ground grass of high livestock is poured with 100 milliliters of 40% ethyl alcohol and insisted in a dark place for 10 days, stirring occasionally. Then, an alcoholic extract from the dried high-crushed grass of high liveliness (tincture) is filtered, the raw materials are squeezed well, the extraction is brought to the initial volume (100 ml) with 40% ethyl alcohol, it is left to stand in a cool place for 3 days, then it is filtered through an anesthetized filter . Received 10% herbal remedy - 10% tincture of high quickness. Store an alcoholic extract from the dried grass of high larkiness (tincture) in a cool dark place.

Example 2. 50 g of dried and ground grass of high liveliness are loaded into five extractors and the first of them is fed from above with 40% ethanol in an amount that creates a layer over the raw material of 5-7 cm, and left for 24 hours. After this period, the liquid from the first extractor is poured into the second, and 40% ethanol is fed into the first extractor until the initial level is obtained. After 24 hours, successive drains are made from the 2nd extractor to the 3rd, from the 1st to the 2nd, and fresh solvent is fed into the first. Every 24 hours, these operations are repeated until the 5th extractor is filled. Then, 100 ml of extract is obtained from the 5th extractor, while simultaneously producing sequential plums of the 1st in the 2nd, from the 2nd to the 3rd, from the 3rd to the 4th, and from the 4th to the 5th th extractor. After an interval of 24 hours, from the 5th extractor with a similar sequence of fluid movement, we obtain in four portions of 100 ml a liquid extract. The resulting extracts are combined (500 ml). Received a herbal remedy - high liquid quickness extract. Store an alcoholic extract from the dried high-larkiness grass (liquid extract) in a cool dark place.

Example 3. 100 g of dried and ground grass of high larkspur is treated with 5% sodium hydroxide solution (pH 9.0) and loaded into a percolator. 12 hours after loading, 2000 ml of chloroform is poured into it. Extraction with chloroform is carried out in a continuous laboratory apparatus consisting of a percolator, a film evaporator and a direct refrigerator. Continuous circulation of the solvent is carried out at a speed of 2 l / h. Duration of exhaustive extraction is 5 days. The concentrated chloroform extract is drained through the evaporator tap. The last portion of the chloroform extract from the percolator is drained through the bottom valve and concentrated in the evaporator. Next, the chloroform extraction is subjected to purification by converting the alkaloids bases into hydrochloric acid salts and, conversely, followed by extraction with suitable solvents. The chloroform extract is then filtered through a desiccant (15 g of anhydrous sodium sulfate). At the last stage, the resulting amount of alkaloids is concentrated on a rotary evaporator under vacuum. A quantitative determination according to [5] showed that the output of the sum of alkaloids from the aerial part of the apex is high was 0.23% in terms of air-dry raw materials. Received herbal remedy containing the amount of alkaloids.

The antihypoxic effect of a liquid extract from dried and ground grass of anthrax with high 40% ethyl alcohol at a ratio of plant materials and extractant 1:10, and a liquid extract from dried and ground grass of ankle with high 40% ethyl alcohol at a ratio of plant materials and extractant 1 : 2, as well as a dry extract of the amount of alkaloids isolated from dried and ground grass of high liveliness, was found thanks to experimental studies.

As a comparison drug used α - tocopherol acetate (vitamin E), belonging to the group of fat-soluble vitamins. This is a basic drug with antioxidant and antihypoxic effects [3]. The prototype was also a reference drug - tincture of stems of elecampane high. Experimental studies of the drug from elecampane high showed a number of positive effects. There is evidence of pronounced antihypoxic activity of this drug in models of single and repeated hypoxic effects [6].

A study of the antihypoxic effect of herbal remedies from the aerial part (grass) of high ankle was carried out on a model of acute tissue hypoxia, which was created by a single intraperitoneal administration of sodium nitroprusside at a dose of 25 mg / kg to animals. The experiments were carried out on outbred male mice weighing 25-30 g. Animals of the first category were obtained from the collection fund of the Research Laboratory of Experimental Biomodeling of the Siberian Scientific Center of the RAMS Scientific Research Institute of Pharmacology (certificate is available). The animals were kept in accordance with the rules adopted by the European Convention for the Protection of Vertebrate Animals used for experimental and scientific purposes (Strasbourg, 1986). Euthanasia was performed in compliance with the "Rules for the Use of Experimental Animals" approved by the Russian Ministry of Health. In the experiment, animals were divided into groups: control, comparisons, experimental. In comparison groups, α-tocopherol acetate at a dose of 100 mg / kg (base preparation) and tincture of Elecampane stems high (prototype) at a dose of 2.5 ml / kg were used as comparison preparations. Control animals received solvent in an equivalent volume. In the experimental groups, the animals received the proposed tool from the dried and ground grass of high larkiness high in different concentrations. All herbal and comparison preparations were administered intragastrically for 5 days, the last time - one hour before the administration of hypoxant. The criterion for evaluating the effectiveness of antihypoxic effects of drugs was the life expectancy of mice.

The results were processed statistically, by the methods of parametric and nonparametric statistics. The differences were considered significantly significant at P _u <0.05 and P _t <0.05 for quantitative characteristics (Wilconson-Mann-Wigney criterion and Student's test, respectively) [7].

An example of a specific implementation of the experiment 1. During the experiment, the proposed tool containing an alcohol extract from the dried ground grass of the grass with high 40% ethanol in the ratio of plant materials and extractant 1:10, in doses of 0.6 and 2.5 ml / kg was administered to experimental animals through a probe into the stomach for 5 days, the last time - one hour before the introduction of sodium nigroprusside. In the comparison groups, α-tocopherol acetate was used at a dose of 100 mg / kg (basic preparation) and tincture of elecampane stems high (prototype) at a dose of 2.5 ml / kg in a similar mode. Control animals received solvent in an equivalent volume. The antihypoxic effect of the drugs was evaluated by the life span of the mice.

As a result of the study, it was shown that a liquid extract from the grass of the larkspur with high 40% ethanol with a ratio of plant materials and extragens of 1:10 in doses of 0.6 and 2.5 ml / kg exhibits a pronounced antihypoxic effect, increasing the life expectancy of mice 2 and 2.5 times, respectively, compared with the control (table 1). The introduction of α-tocopherol acetate 2.3 times increases the time until the onset of signs of the terminal stage when compared with the control. The prototype preparation tincture of elecampane stems increased 1.2 times the life expectancy of mice (Table 1).

Thus, a liquid extract from the grass of the larkspur with high 40% ethanol with a ratio of plant materials and extragens of 1:10 exhibits antihypoxic activity comparable to the action of the comparison drug α-tocopherol acetate (base) and exceeds the prototype drug tincture of stems of elecampane high.

An example of a specific implementation of the experiment 2. During the experiment, the proposed tool containing an alcohol extract from the dried ground grass of the grass with high 40% ethanol in the ratio of plant materials and extragenza 1: 2, in doses of 0.12 and 0.25 ml / kg was administered to experimental animals through a probe into the stomach for 5 days, the last time - one hour before the introduction of sodium nigroproside. In the comparison groups, α-tocopherol acetate was used at a dose of 100 mg / kg (basic preparation) and tincture of elecampane stems high (prototype) at a dose of 2.5 ml / kg in a similar mode. Control animals received the solvent in an equivalent volume. The antihypoxic effect of the drugs was evaluated by the life span of the mice.

An analysis of the obtained data indicates that a liquid extract from the grass of the larkspur with high 40% ethanol with a ratio of plant materials and extractant 1: 2 in doses of 0.12 and 0.25 ml / kg increases the life expectancy of mice by 2 times relative to the control (table 1). The introduction of α-tocopherol acetate 2.3 times, and tinctures of elecampane stems 1.2 times longer before the onset of signs of the terminal stage when compared with the control (Table 1).

Thus, the liquid extract from the grass of the livestock with high 40% ethanol with a ratio of plant materials and extractant 1: 2 exhibits antihypoxic activity comparable to the action of the comparison drug α-tocopherol acetate (base) and surpasses the prototype preparation tincture of high elecampane stems.

An example of a specific experiment 3. During the experiment, the proposed tool containing a dry extract of the amount of alkaloids isolated from dried and ground grass of high larkiness at doses of 0.05 and 1.5 ml / kg was injected into experimental animals through a probe into the stomach for 5 days, the last time - an hour before the introduction of sodium nitroprusside. In the comparison groups, α-tocopherol acetate was used at a dose of 100 mg / kg (basic preparation) and tincture of elecampane stems high (prototype) at a dose of 2.5 ml / kg in a similar mode. Control animals received solvent in an equivalent volume. The antihypoxic effect of the drugs was evaluated by the life span of the mice.

As a result of the experiment, it was found that a dry extract of the amount of alkaloids isolated from dried and ground grass of high larkiness in doses of 0.05 and 1.5 ml / kg increases the life expectancy of mice by 1.6 and 1.7 times, respectively, relative to the control (table .one). The use of α-tocopherol acetate and tincture of stems of Elecampane high 2.3 and 1.2 times, respectively, prolongs the life expectancy of mice (table).

Table

The effect of extracts from the aerial part of the high vivacity (Delphinium elatum L) on the survival of outbred male mice in conditions of acute tissue hypoxia

Group of animals Dose, mg / kg, ml / kg Number of animals Life span, min The control - 10 13.1 ± 0.9 Tincture of stems of elecampane high 2,5 5 15.7 ± 0.2 * The control - 10 9.0 ± 1.2 Tincture 0.6 6 18.9 ± 3.0 * 2,5 6 22.1 ± 1.8 * Extract 0.12 6 17.6 ± 1.8 * 0.25 6 17.0 + 1.6 * Amount of alkaloids 0.05 6 14.8 ± 0.6 * 1,5 6 15.6 ± 1.0 * α-tocopherol acetate 100.0 5 21.1 ± 3.9 * Note: * - the differences are statistically significant compared with the control at Pt, Pu <0.05.

Thus, the dry extract of the amount of alkaloids isolated from the dried and ground grass of high larkiness has a less pronounced antihypoxic effect than the drug for comparison of α-tocopherol acetate (basic), but it is superior in activity to the prototype tincture of stems of elecampane high.

Summarizing the results of the study, it can be noted that all products containing extracts from the dried and ground aerial parts of the livestock have a high antihypoxic effect, with the highest activity in alcoholic extracts from the grass of the livestock with high 40% ethanol in the ratio of plant materials and extragenga 1:10 (tincture) at a dose of 2.5 ml / kg.

Information sources

1. Lukyanchuk VD, Savchenkova L.V. Antihypoxants: state and prospects // Experiment. and clinic. Farmakol. 1998.V. 61. Number 4. S.72-79.

2. Povetieva TN, Latin VG Features of the adaptogenic action of medicinal plants. Tomsk, 2005.P.29.

3. Reference Vidal: Drugs in Russia. M., 2005.S. 143.

4. Zelenskaya K.L., Povetieva T.N., Pashinsky V.G., Fomina T.L., Timina E.A., Perova A.G. Stress-inducing effect of hypoxia of different genesis and its correction by extract from elecampane high // Bull. an experiment. Biology and Medicine 2005. T.139, No. 4. S.406-409.

5. Beshitaishvili L.V., Sultanhodzhaev M.I., Mujiri K.S., Yunusov M.S. Alkaloids of Delphinium tamara // Chemistry of Natural Compounds 1981. No. 2. S.199-201.

6. Zelenskaya K.L., Pashinsky V.G., Povetieva T.N., Nesterova Yu.V. Elecampane high - adaptogen and antihypoxant. Tomsk, 2004.

7. Gubler E.V. Computational methods of analysis and recognition of pathological processes. L., 1978. S. 72-75.

Claims (3)

1. A tool with antihypoxic effect obtained by extraction of dried and ground grass Larkspur with high 40% ethyl alcohol at a ratio of plant material: extractant 1:10. 2. The tool according to claim 1, obtained by extraction of the dried and ground grass of the larkspur with high 40% ethanol in the ratio of plant material: extractant 1: 2. 3. An agent with antihypoxic effect obtained by extraction of dried and ground grass of anthrax with high chloroform and representing the sum of alkaloids.