RU2729784C1 - Herbal tea possessing antioxidant, anti-inflammatory and capillary strengthening properties - Google Patents

Abstract

FIELD: pharmaceutics.SUBSTANCE: invention refers to pharmaceutical industry, namely to an herbal tea with antioxidant, anti-inflammatory and capillary firming activity. Collection possessing antioxidant, anti-inflammatory and capillary firming activity, contains: Aesculus hippocastanum seeds, Bupleurum multinerve herb, Filipendula ulmaria flowers, Aronia melanocarpa fruits, Fragaria vesca leaves, Calendula officinalis flowers in certain proportions.EFFECT: above collection shows manifested antioxidant, anti-inflammatory activity and capillary strengthening.1 cl, 8 tbl

Description

The invention relates to medicine, in particular to herbal products, and can be used, for example, as an antioxidant, anti-inflammatory and capillary-strengthening agent for venous diseases, with increased capillary fragility.

The use of herbal remedies in medical practice is determined by the fact that they are practically non-toxic, contain a complex of biologically active substances and exhibit polyfunctional properties. Charges from medicinal plant materials, as a rule, have a pronounced pharmacological activity in several directions, which is due to the content in their composition of plants containing different groups of biologically active substances. The component composition of the collection, which has a theoretical basis and experimental confirmation, is an effective drug. The use of medicinal products from raw materials of plant origin is noted in various fields of medicine, including diseases of veins and capillaries.

Plants are sources of biologically active substances, and the use of herbal collections allows you to create a composition consisting of a complex of biologically active substances that affect many stages of the pathological process.

Varicose veins currently occupy one of the leading places among cardiovascular diseases and are diagnosed in more than 20% of men and 40% of women [6]. Oxidative stress plays an important role in the development of vascular and capillary diseases; therefore, the use of antioxidant agents at the stage of disease development, its course and for prevention is justified by the use of biologically active substances with an antioxidant effect [1].

To enhance blood flow, increase vascular tone and reduce blood viscosity, biologically active substances of horse chestnut seeds are used, which are triterpene compounds and coumarins [2]. With the development of the pathology of the veins and capillaries, the inflammatory process turns on, therefore it is rational to introduce into the composition of the agent plants with a pronounced anti-inflammatory effect. In case of violation of capillary permeability, drugs with P-vitamin activity are prescribed, which restore the semi-permeability of cell membranes. Such properties are possessed by biologically active substances such as flavonoids - quercetin, rutin, hesperidin. With a combination of biologically active substances - flavonoids and ascorbic acid, synergism is observed in the manifestation of the pharmacological effect of antioxidant and capillary-strengthening.

In medical practice, preparations are used based on biologically active substances of horse chestnut seeds (triterpene saponins and coumarins) - liquid extract, escuzan tablets, escuzan drops. Preparations based on flavonoids: Detralex (contains diosmin and hesperidin), venarus (diosmin and hesperidin), phlebaven (diosmin and hesperidin), phlebodia 600 (diosmin), phlebofa (diosmin), rutin tablets, ascorutin tablets. In most cases, the substances of flavonoids are not of natural origin, but obtained by synthesis [2].

It is known "Antioxidant, capillary-protective, anti-inflammatory and antihistamine agent" for use as an agent with P-vitamin activity, which is an individual compound - dihydroquercetin. [RF patent 2014841, A61K 31/352, AK 61K 36/15. Authors: Sokolov S.Ya., Tyukavkina N.A., Kolkhir V.K. and etc.]. Dihydroquercetin is obtained by extraction of crushed larch wood with water during heating and subsequent purification on a polyamide sorbent.

It is known "Means of plant origin with antioxidant activity" [RF patent 2342942, A61K 36/185, A61P 39/06. Authors: Latypova G.M., Romanova Z.R. and etc.]. The product is a thick extract of spring primrose herb for the prevention and treatment of free radical pathologies.

Known "Collection of medicinal plants antioxidant action" [RF patent 2613171, A61K, authors: Shilova IV, Suslov NI, Samylina IA]. The tool is a vegetable collection from the shoots of the Siberian prince, green leaves of badan, grass meadowsweet.

Known "Composition" Angionorm ", which has antiaggregatory, capillary-protective, venotonic action" [RF patent 2241483, A61K 9/20, A61P 7/00, authors: Kolkhir VK, Voskoboinikova IV]. The herbal composition, which has antiaggregatory, capillaroprotective, venotonic action, is a dry extract obtained by alcohol extraction of a mixture of common horse chestnut fruits, licorice roots, rose hips, hawthorn fruits, taken in certain proportions. The agent is recommended for the treatment of acute pathological conditions and / or exacerbations of chronic diseases associated with vascular disorders, such as increased platelet aggregation, impaired capillary permeability, microcirculation and venous circulation.

The considered funds have disadvantages - an incomplete spectrum of pharmacological activity for the manifestation of antioxidant, anti-inflammatory, capillary-strengthening action.

The closest is "Collecting" Kasmin "for the prevention and treatment of thrombosis" [RF patent 2104024, A61K 36/738, A61K 36/185, authors: Bykov V.A., Klimakhin G.I., Chernykh D.S. and etc.]. The product is a collection of medicinal plants containing horse chestnut seeds (7-12), licorice roots (12-17), peppermint leaves or lemon balm herb (20), hawthorn fruits (20) and the rest of the rose hips. The collection affects not only the homeostasis system itself, but also the cardiovascular system as a whole, lipid and carbohydrate metabolism. The disadvantage of the collection is the insufficient amount of flavonoids in the components of the collection, which determine the antioxidant, anti-inflammatory action and P-vitamin activity.

The purpose of the invention is to develop a herbal collection with antioxidant, anti-inflammatory and capillary-strengthening activity.

The technical result of the invention is the expansion of the range of herbal medicines with antioxidant, anti-inflammatory and capillary-strengthening activity, due to the use of widespread plant raw materials, which have a reliable and secure raw material base, expansion of the spectrum of pharmacological activity due to the multicomponent nature of the drug and the experimentally substantiated ratio of the collection components.

The technical result is achieved by the fact that the plant collection contains in its composition:

grass follicle multifilament - Bupleuri multinervis herba (VFS 42-580-76) - 20% (2 hours);

horse chestnut seeds - Aesculi hippocastani semen (TU 64-4-75-87) - 20% (2 hours);

meadowsweet flowers - Filipendulae ulmariae flores (VFS 42-1777-87) - 30% (3 hours);

chokeberry fruits - Aroniae melanocarpae fructus (FS 2.5.0003.15) - 10% (1 h);

wild strawberry leaves - Fragariae vescae folia (FS.2.5.0016.15) - 10% (1 h);

calendula flowers - Calendulae officinalis flores (FS 2.5.0030.15) - 10% (1 h).

The component composition of the collection and the proposed ratio of plant raw materials were substantiated as a result of theoretical and experimental photochemical and pharmacological studies.

Biologically active substances of the bundle of multistinned flavonoids - rutin, quercetin, isoquercetin, saponins, which normalize capillary permeability and have an anti-inflammatory effect.

The active ingredients of horse chestnut seeds are triterpene saponin escin, coumarins esculin, and fraksin have venotonic, antithrombotic effects, increase venous blood flow, reduce capillary permeability and fragility.

The active ingredients of meadowsweet flowers are flavonoids - quercetin, rutin, kaempferol, narcissin, simple phenolic compounds spirein, isosalicin and monotropitin, caffeic and ellagic acids. They have antioxidant, anti-inflammatory, immunomodulatory effects.

Dry chokeberry fruits are a source of a complex of flavonoids and ascorbic acid. The flavonoids contain cyanidin-3-glucoside, rutin, quercetin, triterpene saponins. Pharmacological action antioxidant, biologically active substances give elasticity and flexibility to blood vessels, restore their permeability, and reduce blood pressure.

Biologically active substances of wild strawberry are ascorbic acid, flavonoids (mainly rutin), traces of essential oil. Aqueous extracts of strawberry leaves have a diuretic effect and are a source of ascorbic acid.

Calendula flowers are a source of carotene, flavonoids narcissin, isorhamnetin, isorhamnetin-3-glucoside, steroids - β-sitosterol, stigmasterol, cholestanol, campestanol; triterpenoids - arnidiol and faradiol, coumarins - scopoletin, umbelliferone and esculetin. The action of the infusion of calendula flowers is anti-inflammatory, antispasmodic, improving regeneration processes.

The technology of preparation of the collection is that each type of medicinal raw material is crushed separately: grass and leaves to a particle size of no more than 7 mm, flowers no more than 5 mm, fruits - no more than 3 mm, seeds - no more than 2 mm. Then the required amount of ingredients is weighed, thoroughly mixed until a uniform mixture is obtained and sifted from dust through a sieve with a hole diameter of 0.18 mm.

To prepare the infusion, 10 g of the collection is poured into 210 ml (taking into account the calculated water absorption coefficient equal to 2.1) with purified room temperature water, infused in an infundir apparatus in a boiling water bath with frequent stirring for 15 minutes, then cooled at room temperature for 45 minutes ; filter (squeezing the plant material) and add water to the required extraction volume. As a result, an infusion is obtained in the ratio of raw materials - extractant 1:10 [3].

Qualitative analytical reactions and chromatographic studies in the proposed collection revealed triterpene saponins escin, saicosaponin A, flavonoids - rutin, hyperoside, quercetin, isorhamnetin, isorhamnetin-3-O-rutinoside, cyanidin-3-glucoside; phenol carboxylic acids - 5-O-caffeylquinic acid, 1,3-di-O-caffeylquinic acid; coumarins - esculin, esculetin; tannins.

The quantitative content of the sum of flavonoids was determined by the spectrophotometric method based on the reaction of complexation of flavonoids with aluminum chloride in terms of rutin. The content of the sum of phenolcarboxylic acids in terms of chlorogenic acid was determined by the spectrophotometric method, the content of ascorbic acid by the titrimetric method (recommended for rose hips according to the GF XIV edition). The content of the sum of polyphenolic compounds was determined by the permanganatometric method, extractives according to SP XIV [3].

To determine the optimal ratio of ingredients in the collection were prepared options for collections No. 1-No. 5 with a different ratio of components (table. 1).

Further, by the method of maceration, 50% ethyl alcohol was prepared from the extracts in the ratio of raw material-extractant (2:50), in which the content of phenolic compounds, dry residue and antioxidant activity were determined (Tables 2 and 3).

Antioxidant activity was studied using donor blood serum in in vitro experiments. Method: to 2 ml of donor blood serum was added 1 ml of dealcoholized test extract, 0.1 ml of 0.5 M copper (II) sulfate solution, as an inducer of lipid peroxidation. For comparison, a control without test extraction was used. The reaction mixture was incubated for 24 hours at 37 ° C, and examined for the content of diene conjugates (DC) and malondialdehyde (MDA) [9].

All collection options reduce the formation of DC in comparison with the control (purified water) from 10.50 to 37.00%. Extraction of collection option No. 3 maximally reduces the content of DC by 37% and MDA in by 67.33% (Table 2).

Extracts from collection No. 3 contain the largest amount of biologically active substances and have high antioxidant activity. We have chosen this ratio of components as the base for compiling the recipe for the collection.

Analysis of plant raw materials from collection No. 3 showed the content of the sum of polyphenolic compounds - 11.96 ± 0.42%, the sum of flavonoids in terms of rutin 4.24 ± 0.13%, the amount of phenol carboxylic acids in terms of chlorogenic acid 3.50 ± 0, 06%, ascorbic acid - 0.25 ± 0.03%, extractives extracted with purified water - 33.01 ± 0.4%, extractives extracted with ethyl alcohol 50% - 43.60 ± 0.8%.

Pharmacological studies were carried out in accordance with the established documents "On the approval of the rules of laboratory practice" (Ministry of Health of the Russian Federation, order dated June 19, 2003 No. 267), Good Laboratory Practice for Nonclinical Laboratory Studies (FDA, 21 CFR Part 58, 22.12.1978), OECD Principles on Good Laboratory Practice (OECD, ENV / MC / CHEM (98) 17, 1977).

Study of anti-inflammatory activity of collection No. 3. The experiments were carried out on 30 white Wistar rats of both sexes weighing 180-200 g. The studies were carried out in three series of experiments: evaluation of the effect on the exudative phase of inflammation [7], the phase of proliferation [8] and on the processes of alteration [5]. In all experiments, animals were enterally injected with infusion of collection No. 3 at a dose of 100 mg / kg (the first group of rats) and in the form of a reference drug - an infusion of chamomile flowers at a dose of 100 mg / kg (the second group of rats). Animals of the control group were injected with water in an equal volume according to a similar scheme. In the first series of experiments, the antiexudative effect of collection No. 3 was studied on a model of acute inflammatory edema caused by subplantar injection of 0.1 ml of 2% formalin aqueous solution into the hind paw of rats. The infusions of collection No. 3 and chamomile were administered to the corresponding groups of animals 2 hours before the introduction and 5 and 18 hours after the introduction of formalin. Measurement of the volume of the paws was carried out using an oncometer before the start of the experiment and at the moment of maximum development of edema (after 4 hours). Collection activity No. 3 was expressed as a percentage of edema inhibition.

In the second series of experiments, the effect of collection No. 3 on the processes of alteration and regeneration in the focus of inflammation was studied by subcutaneous injection of 0.5 ml of a 9% solution of acetic acid into the back region of rats with previously trimmed hair. At the same time, rats were injected with a dextran solution intraperitoneally at a dose of 300 mg / kg. The first administration of the collection preparation No. 3 and chamomile to the indicated groups of rats was carried out 1 hour before the administration of acetic acid, and then every day once a day for 25 days. On days 9 and 29 of the experiment, the area of necrotic tissue was assessed by drawing a necrosis contour on a transparent film.

In the third series of experiments, the effect of collection No. 3 on proliferation processes was studied by implanting sterile cotton balls weighing 15 mg under the skin of rats in the back in aseptic conditions. Infusion collection No. 3 to the first group of rats and chamomile pharmacy to another group of rats was injected intragastrically 1 time per day for 7 days. After 7 days, granulomas were removed from animals anesthetized under light ether anesthesia and immediately weighed on an analytical balance and after drying at 70 ° C for 24 hours (to constant weight).

Statistical data processing was carried out using Student's t-test.

As a result of the studies, it was found that collection No. 3 reduces formalin edema of rat paws by 38.60%, while pharmaceutical chamomile - by 29.80% compared to control (Table 4).

In addition, collection # 3 reduced tissue damage. In particular, on the 9th day in rats receiving infusion of collection No. 3, the area of necrotic tissue was 35.00% less than in rats of the control group, and in animals receiving infusion of chamomile, by 39.20%. Collection No. 3 effectively stimulated regenerative processes, which resulted in a decrease in the area of alteration on the 29th day of the experiment by 58.90%, while in rats receiving an infusion of chamomile, the area of damage decreased by 27.10% (Table 5).

As a result of the third series of experiments, it was found that in animals that received collection No. 3, the mass of the extracted granulomas exceeded the control by 21%. (Table 6). The infusion of chamomile used as a comparison, on the contrary, reduced the mass of granulomas by 45% compared to the control.

Collection number 3 reduces formalin edema, altration and promotes the process of proliferation. This proves the pronounced anti-inflammatory activity of collection No. 3.

Study of the effect of collection No. 3 on capillary permeability. To study the capillary-strengthening effect, we used a model of local inflammatory reaction using xylene on female rats weighing 180-200 g according to the method of K.N. Monakova [4]. The animals were divided into 3 groups of 6 animals each. The first group is intact animals. The second group received rutin solution as a reference drug. The third group was injected with the infusion of collection No. 3 30 minutes before the experiment. In rats, the hair on the skin of the abdomen was cut (0.5 cm area). The drugs were administered per os in a volume of 3 ml (100 mg / kg dry matter) 1 hour before the introduction of a permeability indicator, the role of which was played by a solution of trypan blue. Trypan blue was injected into the tail vein of rats in the form of a 0.5% solution in 0.9% sodium chloride solution at the rate of 1 ml per 100 g of animal weight. After 30 s, a drop of xylene was applied to the skin of the anterior abdominal wall. The time of appearance of petechiae and their distinct staining was recorded. The time of appearance of blue-colored spots on the skin and their diameter served as an indicator of capillary permeability.

The experimental results were statistically processed by methods for a small sample with the determination of the average value (M) and error (m). The degree of reliability of the research results (P) was assessed using the parametric Student's t test for a small sample. Differences between control and experimental data were considered significant at a probability of 95% (P≤0.05).

The difference in the time of the appearance of spots and their diameter before and after the administration of the preparations was used to judge their effect on the permeability of the capillaries.

As can be seen from table 7, in the control, the time of appearance of petechiae averaged 76 seconds; clear staining time 110 sec. We took these indicators as 100%. In the group of animals treated with rutin solution, the time of appearance of petechiae was 111 sec and the time of distinct staining was 126 sec. With the introduction of collection infusion No. 3, these indicators were respectively 126 seconds and 138 seconds, relative to the control. In percentage terms, it looks like this. The rutin solution increased the time of appearance of petechiae by 46.1% and the time of distinct staining by 12.7%, while in the group receiving infusion of collection No. 3, these indicators were 65.7% and 25.5%, respectively.

Collection No. 3 surpasses rutin in capillary-strengthening effect and collection "Kasmin" (Table 7). In terms of the content of the main biologically active substances responsible for the antioxidant, anti-inflammatory and capillary-strengthening effect, collection No. 3 surpasses the collection of "Kasmin" (Table 8).

Literature

1. Glazkov A.A. The role of oxidative stress in vascular damage in arterial hypertension // Scientific community of students of the XXI century. Natural Sciences. Electronic collection of articles based on the materials of the L student international scientific and practical conference. - Novosibirsk: Ed. ANS "SibAK". - 2017. - No. 3 (49) - S. 36-34. / [Electronic resource] - Access mode. - URL: http://www.sibac.info/archive/nature/3(49).pdf

2. State Register of Medicines as of 09/02/2019 [Electronic resource] URL: http://grls.rosminzdrav.ru

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4. Zhilina, I.V. Stepanova E.F., Ogurtsov Yu.A., Golova G.A. Study of the capillary-strengthening action of the gel with the extract of meadowsweet // Scientific statements. Medicine series. Pharmacy. - 2010. No. 22 (93). Issue 12 (2). - from. 29-31.

5. Oyvin I.A., Shetel S.L. Methods for studying local capillary permeability disorders // Mater, on the pathogenesis of inflammation and pathology of blood proteins. - Dushanbe, 1961. - 49. - No. 5. - 167-173.

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Claims (1)

Herbal collection, which has antioxidant, anti-inflammatory and capillary-strengthening effect, containing the seeds of the common horse chestnut, characterized in that it additionally contains the herb of the multifilament buckwheat, the meadowsweet flowers, the chokeberry fruits, the leaves of wild strawberries, the calendula flowers with the ratio of the components of the collection in parts, respectively 2 : 2: 3: 1: 1: 1.