RU2637414C1 - Method for production of preparation with anti-inflammatory activity - Google Patents

Abstract

FIELD: pharmacology.

SUBSTANCE: method for production of an agent having anti-inflammatory activity, including garden burnet triple extraction with ethyl alcohol, filtering, evaporation, concentration and drying of the alcohol extract, at that, garden burnet herb crushed to a particle size of 2.0 mm is used, extraction is done by 70% alcohol at 90°C at raw materials - extractant ratio of 1:50 within 2 h, at that, this first extraction is performed for 60 minutes, the second and the third - for 30 minutes.

EFFECT: increased anti-inflammatory activity of the agent.

5 tbl, 2 ex

Description

The present invention relates to medicine, namely to pharmacy, and relates to a method of obtaining funds from plant materials with anti-inflammatory activity.

Currently, there are a large number of anti-inflammatory drugs and numerous studies are underway to develop, study and put into practice drugs for the treatment and prevention of inflammatory diseases. The most widely represented group of non-steroidal anti-inflammatory drugs, for example diclofenac, indomethacin, ibuprofen, acetylsalicylic acid and others, which, with their pronounced therapeutic effect, are not devoid of side effects, have contraindications and restrictions on their use, namely, they cause immunosuppression, hemorrhagic complications, and an increased risk of ulcerative diseases of the stomach and duodenum and its complications [Pharmacology: textbook. - 10th ed., Rev., Rev. and add. - Kharkevich D.A. 2010. - 752 p .; Karateev A.E., Nasonova V.A. The problem of NSAID-induced gastropathy: past and present // breast cancer. The appendix "Diseases of the digestive system." - 2004. - No. 1. - S. 36-43; Schacke N., Docke W. D., Asadullah K., Mechanisms involved in the side effects of glucocorticoids // Pharmacology & therapeutics. - 2002. - T. 96. - No. 1. - C. 23-43].

Expanding research to find sources to obtain new effective and safe medicines, including herbal origin, is an urgent task of medicine. Of great interest in this direction are medicinal plants as potential sources of new anti-inflammatory drugs. The advantage of medicinal plants is a wide range of their therapeutic and prophylactic effects, low toxicity, mild action, the ability to quickly eliminate symptoms of exacerbation, prevent relapses [Mashkovsky, M.D. Medicines / M.D. Mashkovsky. - 15th ed., Rev., Rev. and add. - M .: LLC "Publishing house New wave", 2005. - 1200 p.].

Known herbal remedy "Kaleflon" containing an extract of calendula flowers, which is used as anti-inflammatory, stimulating reparative processes; herbal remedy "Romazulan", which is a liquid extract from chamomile flowers pharmacy and used as an anti-inflammatory, antiseptic, antispasmodic [Reference Vidal. Medicines, 2016]. The disadvantages of these funds are the possibility of developing side effects in the form of allergic reactions with intolerance to the components, a burning sensation and pain in the epigastric region, bitterness in the mouth.

Herbal preparations based on individual components isolated from plants are known, for example, the use of the ursolic acid derivative as an agent with anti-inflammatory, antioxidant, hepatoprotective properties, which exhibits high biological activity, however, the disadvantage of this compound is its low solubility in polar and non-polar solvents, which can lead to low bioavailability [RF Patent No. 2436793 "A tool with antioxidant, hepatoprotective and p otivovospalitelnymi properties ", A. Sorokin et al., 12.20.2011]. A dihydroquercetin-based agent with anti-inflammatory, capillary-strengthening, antioxidant and antihistamine effects. However, this compound is not water soluble, which negatively affects the bioavailability [RF Patent No. 20144841 "Method for the preparation of a soluble dihydroquercitin preparation", Sokolov et al., 30.06.1994].

A known method of obtaining funds with anti-inflammatory and wound healing activity, including extraction with ethanol of alfalfa grass, which is carried out three times, sequentially in three percolators at a ratio of raw materials: extractant 1: 2 at room temperature, the initial infusion in each percolator is carried out for 6-7 hours, the combined extracts are defended and filtered [application RU 97105729, published on 04/20/1999].

A known method of obtaining funds with anti-inflammatory activity, by extracting plant material, while the herbal material used is the collection of medicinal plants, consisting of the aerial part of the woolly panzerina and speckled cocoa, which is extracted with 50-60% ethyl alcohol with triple extraction in the ratio plant material - extractant equal to 1: (14-16) (taking into account the coefficient of water absorption), the first and second time for 60 minutes, the third - 30 minutes at a temperature of 60-65 ° C [application RU 201 3114909, published 10/10/2014].

A known method of obtaining funds with anti-inflammatory activity, by extracting a plant mixture containing, by weight, by weight: black leaves of frangipani - 3.0; chamomile flowers pharmacy - 2.8; yarrow grass - 2.4; peppermint leaves - 1.8. The plant mixture, ground to a particle size of 0.5-2.0 mm, is extracted twice with 80% ethanol at room temperature with a feed: extractant ratio of 1: (10-12) for 2 hours, then twice with 40% ethanol at a temperature of 90 ± 5 ° C for 1 h, the filtered alcoholic extracts are evaporated, combined, concentrated, dried in a vacuum oven to obtain a dry extract and crushed [patent RU 2331429, 2008].

The closest analogue of the invention is a method of obtaining funds with choleretic and anti-inflammatory activity, which consists in the fact that the rhizomes and roots of elecampane high, the rhizomes and roots of the Sophora yellowish, the rhizomes and roots of the hemorrhagic medicinal, the rhizomes of ginger medicinal, Siberian elderberry, the blood of the hawthorn red and apple fruit are extracted with 25-30% ethanol in the ratio of raw materials - extractant equal to 1: (18-20), at 60-65 ° C with constant stirring. The process is repeated three times. The extraction time at the 1st phase contact is 120 minutes, at the 2nd and 3rd phase contacts - 60 minutes. The extracts are filtered, combined, evaporated and purified by separation. The purified extract is doubled, dried in a vacuum dryer and crushed [patent RU 2159123, 2000]. The disadvantages of this method are prolonged extraction, in which biologically active substances can partially be destroyed, as well as the use of a complex of medicinal plants with versatile pharmacological effects, which can interfere with the allocation of the main group of tannins, which are contained in the drug hemorrhage to a greater extent and provide a greater anti-inflammatory effect.

The objective of the invention is to expand the range of anti-inflammatory drugs based on medicinal plants.

The technical result is an increase in anti-inflammatory activity, a simplification of the method due to the use of one type of medicinal plant, containing tannins to the maximum, which, when interacting with an inflamed surface, provides its compaction and an active reduction in the area of the inflammatory process.

The specified technical result is achieved by the fact that in the method of obtaining funds having anti-inflammatory activity, including triple extraction with ethyl alcohol hemorrhage drug, filtration, evaporation, concentration and drying of the alcohol extract, according to the invention, the herb hemorrhage medicinal, crushed to a particle size of 2.0 mm, is used, extraction is carried out with 70% alcohol at 90 ° C with a ratio of raw material - extractant 1:50 for 2 hours, while the first extraction is carried out for 60 minutes, the second and a third th - 30 min.

The proposed method of obtaining the sum of extractive substances is quite simple, does not require laborious costs, allows you to get a product of constant composition.

To prove the materiality of the proposed conditions for a method for producing an agent having anti-inflammatory activity, namely, the concentration of extractant, the degree of grinding of raw materials, the ratio of raw materials and extractant, the duration and multiplicity of the extraction process, a series of experiments were carried out in comparison with the known and used raw blood hemoptysis raw material - rhizomes and roots ( Rhizoma et radices Sanguisorbae ), the data of which are presented in tables 1-4.

One of the quality indicators of medicinal plant materials that characterizes the content of the whole complex of substances (active, associated and ballast) is the determination of the amount of extractive substances, which was carried out according to the pharmacopeia method [State Pharmacopoeia of the Russian Federation XIII edition, OFS.1.5.3.0006.15]. When studying this indicator, various extractants were used (purified water and ethyl alcohol in a concentration of 30 to 96%) in order to determine which of them most fully extracts extractives (Table 1). It was found that the optimal extractants are 70% ethyl alcohol and water (at 100 ° C), which allow to extract the largest amount of extractive substances.

The peculiarity of extracting biologically active substances from medicinal plant materials is that the cell wall, which is a semi-permeable septum, is on the way to the substances contained in the cell. Therefore, the degree of its fineness is of particular importance for the extraction of active substances from plant material. To assess the efficiency of extraction, the content of extractives was determined (Table 2). It was found that grinding raw materials to a particle size of more than 2.0 mm is undesirable due to a decrease in the yield of extractive substances.

The yield of extractives is affected by the ratio of raw materials - extractant. Experiments were carried out to determine the optimal ratio of raw materials to extractant (table. 3). As a result of the analysis, it was found that the ratio of 1:50 is optimal, a further increase is impractical, since with a significant increase in the volume of extractant, the content of extractives does not significantly increase.

The effect of time and extraction rate on the yield of the sum of biologically active substances was studied (Table 4). Based on the experiments, it was found that the equilibrium state in the system for the dominant components at the first contact of the phases is achieved in 60 minutes, at the second and third contact of the phases - in 30 minutes. Therefore, triple extraction ensures the depletion of raw materials for the main components by an average of 80-90% of the initial content in the raw materials.

As a result of the analysis, it was found that the optimal conditions for extracting the herb of the hemorrhagic drug are: extractant - 70% ethyl alcohol, its ratio to raw materials is 1:50, the degree of grinding of the raw materials is 2.0 mm, the extraction temperature is 90 ° C, the time extraction - 60 minutes, the second - 30 minutes, the third - 30 minutes

The method is illustrated by the following examples:

EXAMPLE 1 1.0 g (accurately weighed) milled medicinal plants burnet (Herba Sanguisorbae, Sanguisorba officinalis L., Rosaceae), sieved through a sieve with apertures of 2 mm, placed in a conical flask of 250 ml, 50 ml of 70 % ethyl alcohol, the flask is closed with a stopper, weighed with an accuracy of ± 0.01 g and left for 1 hour. Then the flask is combined with a reflux condenser and heated in a water bath, maintaining a weak boil for 1 hour. After cooling, the flask with the contents again close with the same cork ivayut and make up for the loss in mass of solvent. The contents of the flask are filtered through a dry paper filter into a volumetric flask with a capacity of 150 ml. The filter with a weighed portion is again placed in the original flask and the boiling procedure is repeated in a water bath under reflux for another 2 times for 30 minutes, each time filtering the extract into the same flask and adjust the solution to the mark with the same solvent. The combined alcohol extract was evaporated on a rotary evaporator to remove alcohol. The final product is obtained by drying the concentrated extract in a vacuum oven.

The preparation of aqueous extraction from the raw material of the hemorrhage was carried out in accordance with the requirements of the State Pharmacopoeia of the Russian Federation of the XIII edition, article “Infusions and decoctions” (OFS.1.4.1.0018.15).

Example 2. 10.0 g (accurately weighed) of the chopped medicinal plant material of a blood hemorrhage sifted through a 2 mm sieve, placed in an infundir apparatus, filled with 100 ml of purified room temperature water, insisted on a boiling water bath for 15 min ( grass) and for 30 minutes (rhizomes and roots), then the grass is cooled at room temperature for 45 minutes, and the rhizomes and roots are filtered hot to avoid precipitation of tannins in the sediment, squeeze the plant material and add water to the required emogo extraction volume.

For a comparative assessment of the anti-inflammatory activity of water and alcohol extracts from the raw materials of the hemoptysis, a screening model of formalin paw edema in mice was used [Khabriev R.U. Guidance on the experimental (preclinical) study of new pharmacological substances / R.U. Khabriev. - M .: Medicine, 2005. - 832 p.]. The studies were carried out on 36 adult white mice - males weighing 19-25 g. For the experiments, animals with an average motor activity prevailing in the population were selected, and therefore it can be argued that these mice will develop the most typical reaction to any effect. The experimental animals were kept in vivarium conditions (with natural lighting; at a temperature of 22-24 ° C) and free access to water and food. Before the experiment, animals were quarantined for 10-14 days. Then the animals were injected intragastrically using a special extraction probe prepared from grass, rhizomes and roots of the blood fever in an amount of 0.08-0.1 ml in a preventive mode for 7 days (the dried alcohol extract was dissolved in water in the ratio 1 before administration: 10). The control group received water. As a comparison drug used diclofenac-sodium "Hemofarm AD" at a dose of 10 mg / kg On the last day, an hour after the administration of the extracts, phlogogen was administered. Acute inflammation was induced by introducing 0.05 ml of a 1% formalin solution under the plantar aponeurosis of the right leg of the mouse. This caused paw swelling that develops over time. After 5 hours, the animals were euthanized under ether anesthesia, observing the "Rules for working with laboratory animals" (Appendix to the order of the Ministry of Health of the USSR dated 08/12/1977 No. 755). The anti-inflammatory effect was evaluated by reducing the swelling of the legs and performed statistical processing of the results of the study (table. 5). [Glanz S. Biomedical statistics / S. Glanz. - M.: Practice, 1998. - 459 p.].

As a result, it was found that all the tested extracts of the blood hemorrhage during intragastric administration showed anti-inflammatory activity, but it was more pronounced for alcoholic extracts than water, and given the belonging to the morphological group of raw materials, a more pronounced effect was observed in grass compared with rhizomes and roots. So, when using extracts from the herb of the hemorrhagic drug, the swelling of the legs in mice significantly decreased by 26.7% when using an aqueous solution and by 33.3% when using an alcohol extract. When extracts from rhizomes and roots were used, the swelling of the legs decreased to a lesser extent: by 10% when using an aqueous solution and by 14.4% when using an alcohol extract.

Claims (1)

The method of obtaining funds with anti-inflammatory activity, including triple extraction with ethyl alcohol hemorrhage drug, filtering, evaporating, concentrating and drying the alcohol extract, characterized in that they use medicinal hemorrhage grass, crushed to a particle size of 2.0 mm, the extraction is carried out 70% alcohol at 90 ° C at a ratio of raw materials - extractant 1:50 for 2 hours, with the first extraction carried out for 60 minutes, the second and third for 30 minutes.