RU2619207C1 - Biologically active food supplement with cancer-preventive action - Google Patents

Abstract

FIELD: pharmacology.

SUBSTANCE: drug with a cancer-protective effect, containing a complex extract from green tea leaves, turmeric roots, black cumin seeds, Japan ampelopsis rootgrass, Sigesbeckia orientalis grass, Baikal skullcap roots, leaves and twigs of crosswise-opposite microbiota Natural as components of plant origin, further containing natural honey, at a certain component ratio.

EFFECT: above described agent has a pronounced protective effect, it allows to increase body adaptive potential and resistance to onset and development of neoplastic diseases, and also expands the arsenal of cancer-protective means.

4 dwg, 1 tbl, 8 ex

Description

The invention relates to cancer preventive agents containing substances of plant origin, intended for prophylaxis and adjuvant therapy in order to reduce the risk of the onset and development of cancer.

The spread of tumor diseases is one of the most acute problems of modern healthcare. Cancer (malignant neoplastic disease) is one of the leading causes of death in the world. According to WHO, oncology currently accounts for 13% of the total number of deaths. Over the past 25 years, the incidence of cancer has increased 1.5-2.0 times, and by 2030 it is projected to increase threefold, with a particularly alarming situation in developed countries. Many common types of cancer have one thing in common - these diseases are extremely difficult to treat with fully existing anticancer therapeutic agents. It should be recognized that the treatment of cancer is currently highly costly and relatively ineffective. At the same time, it is believed that up to 40% of cases of cancer can be prevented with a healthy diet, physical activity, and abstinence from tobacco use. Prevention may include the use of various products and agents containing substances that prevent the degeneration of normal cells into cancer cells.

Well-known carcinogenic agents are polyphenols from green tea, flavonoids from berries, resveratrol from red grapes, capsaicin from peppers, curcumin from a tropical turmeric plant and many others [Lu Y-P, Lou Y-R, et al. (2000) Stimulatory effect of oral administration of green tea or caffeine on ultraviolet light-induced increases in epidermal wild-type p53, p21 (WAFI / CIPI), and apoptotic sunburn cells in SKH-1 mice. Cancer Res 60: 4785-4791; Samaha H.S., Kelloff G.J., et al. (1997) Modulation of apoptosis by sulindac, curcumin, phenylethyl-3-methylcaffeate and 6-phenylhexyl isothiocyanate apoptotic index as a biomarker in colon cancer chemoprevention and promotion. Cancer Res 57: 1301-1305; Yang K., Lamprecht S. A., et al. (2000) Chemoprevention studies on the flavonoids quercetin and rutin in normal and azoxymethane-treated mouse colon. Carcinogenesis 21: 1655-1660; Tanaka, T., Kohno H., et al. (2002) Modifying effects of dietary capsaicin and rotenone on 4-nitroquinoline-1-oxide-induced rat tongue carcinogenesis. Carcinogenesis 23: 1361-1367].

Oncological prophylactic agents from algae are known, such as “Klamin” containing saponified kelp concentrate [RU 2034560 C1, 05/10/1995], “Fitolon” and “Fitolon-M” containing ethyl alcohol, copper derivatives of chlorophyll, food flavoring and water [RU 2031654 C1, 03/27/1995], "Fucus" containing a fucus extract obtained from vesiculus fucus [RU 2116798 C1, 08/10/1998].

However, not one of the known biologically active compositions exhibits an enhanced carcinogenic effect.

As a prototype, an agent for the prophylaxis of oncological and non-oncological diseases was selected, which is a dry water-alcohol extract of individual plants selected from the group consisting of geranium, plantain, calendula, shrubby cinquefoil and angelica, or a mixture thereof [RU 2208446 C2, 07.20.2003 ]. A study of the effect of extracts on the growth of chemically induced tumors showed their pronounced chemoprophylactic effect.

A disadvantage of the known plant composition is the relatively weak effectiveness of the tests when tested in vivo using experimental animal models of carcinogenesis.

The technical task of the present invention is to expand the arsenal of tools with cancer-preventing properties.

The problem has been solved by creating a new agent with a cancer-preventive effect, including components of plant origin, which according to the invention as a component of plant origin contains a complex extract of green tea leaves, turmeric roots, seeds of black corms, root tubers of the Japanese vineyard, sisezbekia east grass, roots of Scutellaria baicalensis, leaves and branches of microbiota are cross-paired and additionally contains natural honey, in the following ratio of ingredients in, kg per 100 gave:

Natural honey 600.0-675.0 Green Tea Leaf Complex Extract, turmeric roots, seeds of Chernushka sowing, root of a Japanese vineyard, sigezbekia eastern herbs, roots of Scutellaria baicalensis, leaves and branches of microbiota cross-paired 57.0-60.0

in order to obtain a complex extract, ethyl alcohol of 35% or 70% concentration is used, and the complex extract contains ingredients in the following ratio, kg per 100 decaliters of extract:

Green tea leaves 24.0-25.0 Turmeric roots 24.0-25.0 Seeds of Chernushka sowing 24.0-25.0 Japanese vineyard root tubers 24.0-25.0 Sigezbekia eastern grass 24.0-25.0 The roots of Scutellaria baicalensis 24.0-25.0 Leaves and branches of the microbiota cross-paired 24.0-25.0 Ethyl alcohol, 35% or 280.0-300.0 Ethyl alcohol, 70% 570.0-590.0 Distilled water rest

It is not known from the prior art that the agent of the claimed composition has a cancer-preventive effect, which allows you to enhance the cancer-preventive effect on the in vivo model of skin carcinogenesis, enhance the adaptive and adaptive potential and resistance of the body to the onset and development of tumor diseases, and also expand the arsenal of drugs that have cancer-preventive properties.

The inventive tool has a significant modulating effect on immunological processes in the body in such a way that it stimulates its oncoprotective properties; leads to the correction of morphological and biochemical parameters towards their normalization during the development of skin carcinogenesis; helps to reduce the incidence of new tumor foci, the growth rate, the number and size of tumors in animals; improves the clinical picture of the manifestation of oncopathology, significantly increasing the latent period of tumor growth.

The components of the proposed funds in the claimed qualitative and quantitative ratio exhibit synergistic properties, providing its cancer-preventive effect.

The chemical composition of green tea leaves is represented by various compounds, such as dry weight proteins (15-20%), amino acids (1-4%), carbohydrates, lipids, sterols, vitamins, caffeine and theophylline, minerals, polyphenols - catechins. The predominant importance in ensuring the medicinal properties of green tea belongs to epigallocatechin (EGCG). It has been shown that green tea catechins have a multi-targeting effect, inhibit the proliferation and metastasis of tumor cells, inhibit angiogenesis, induce apoptosis, and reduce oxidative damage to metabolites.

Turmeric contains vitamins K, B, B1, B3, B2, C and trace elements: calcium, iron, phosphorus and iodine. However, turmeric contains components that even in microscopic amounts have a healing effect on the human body. These are essential oils and their constituent sabinen, borneol, zingibern, terpene alcohols, fellandren, curcumin and a number of other components. Curcumin occupies a special place in this list. The useful properties of curcumin have long been interested in doctors. In the course of scientific experiments, it turned out that curcumin causes the death of pathological tumor cells, without affecting healthy cells. Thus, the use of drugs containing curcumin not only suspended growth, but also prevented the emergence of new malignant tumors.

Seeds of Chernushka sowing seeds contain semi-drying fatty oil (up to 44%), glyantide melantin, essential oil. It is traditionally used for various procedures related to problems of the respiratory system, stomach, intestines, kidneys, liver, cardiovascular system, increasing immunity and improving the general condition of the body. In folk medicine, seeds of black nigella were used as tea as a diuretic, carminative, choleretic, laxative, lactogenic and anthelmintic.

The root tubers of the Japanese vineyard contain polyphenols of both flavonoid and non-flavonoid nature. Among the main flavonoids, quercetin and its derivatives, anthocyanins and their derivatives, (+) catechin, (-) epicatechin and their derivatives, rutin can be noted. Of the main non-flavonoids, gallic acid, transcofeic and transcumaric acids, as well as a stilbene derivative, resveratrol, were found in significant quantities. Among polyphenols of grapevines, (+) catechin, (-) epicatechin, oligomeric and polymeric compounds of these substances, quercetin, gallic and caffeic acids possess the maximum antioxidant activity. A decoction of the roots is used as a painkiller, antitumor and blood purifying agent for syphilis, ulcers, carbuncles, abscesses, burns and pyoderma.

Sigezbekia eastern herb has valuable healing properties, which are caused by the content of terpenoids, 3-7-dimethyl ester quercetin, diterpenoids, orientalide sesquiterpenoids and orientin, as well as stigmasterol and beta-sitosterol steroids. A decoction based on eastern sigezbekia is used for various malignant tumors, it is recommended to accelerate the healing of wounds and internal injuries, including with peptic ulcer of the stomach and intestines. The components of the herb sigezbekia east contribute to the regulation of the immune system and are characterized by antirheumatic, antiseptic, antispasmodic properties.

The roots and rhizomes of Scutellaria baicalensis contain flavonoids, valuable in their therapeutic effect, the most important of which is baicalin, which has an anticonvulsant effect and has a positive effect on the vessels, expanding them. Also important role in the complex of active substances of Scutellaria is played by scutellarein, which protects against the action of toxins, and vogonin, which has a vasoconstrictor effect. The general effect of flavonoids helps protect against viruses. Coumarins in the composition of Scutellaria have an antitumor effect. Scutellaria roots are also distinguished by a significant concentration of pyrocatechins, minerals, resins and tannins. The plant contains starch and essential oil. The whole complex of active components of Scutellaria has the ability to relieve cramps, provide a calming effect and reduce blood pressure. Another amazing property of this plant is its ability to inhibit the formation of metastases. The use in animals has confirmed the ability of this plant to prevent the further development of tumors, coupled with the regulation of the level of white blood cells in the blood.

Leaves and branches of the cross-paired microbiota (relative of the thuja eastern) contain many essential oils and resins, as well as bioflavones. The extract of this plant has an antifungal effect. It is known to use this plant for the treatment of colds, as an expectorant for pneumonia and bronchitis.

The use of honey as an effective medicine is based on many of its properties, including antibacterial, bactericidal, anti-inflammatory and anti-allergic effects. The therapeutic effect of honey is facilitated by the composition of sugars, minerals, trace elements, vitamins, enzymes, biologically active substances. Honey is used as a restorative, tonic, restorative remedy. It is used for diseases of the cardiovascular system, kidneys, liver, biliary tract, gastrointestinal tract. When used in food, honey is quickly absorbed by the body. In addition, honey contains a large number of aromatic substances that improve the taste of the product.

The proposed tool is obtained as follows.

The estimated quantity of green tea leaves, turmeric root powder, sowing black root seeds, Japanese root tuber, sisezbekia herb Oriental, Baikal scutellaria roots, branches and leaves of the microbiota cross-pair are placed in the container. The calculated amount of 35% or 70% ethyl alcohol of higher purity is poured to the plant material and insisted for 10 days at room temperature with periodic stirring. The resulting tincture is filtered through 5-6 layers of gauze and evaporated on a rotary evaporator.

The calculated amount of distilled water and natural honey preheated at 60 ° C is added to the plant complex extract and mixed until a homogeneous state.

A study of the chemical composition of plant extracts designated G35 and G75 by UV absorption spectra (standard is quercetin) showed that the flavonoid content in samples G35 and G70 is 48.97 and 62.13 mg / ml, respectively. 20 amino acids were also found in the extracts (Biochrom-30 amino acid analyzer), such as aspartic and glutamic acids, proline, alanine, arginine, etc.

The possibility of carrying out the claimed invention is illustrated by examples.

Example 1. For the preparation of drugs with a cancer-preventive effect, the consumption of constituent ingredients per 100 decalitres, kg:

Natural honey 600,0 Green Tea Leaf Complex Extract, turmeric roots, seeds of Chernushka sowing, root of a Japanese vineyard, sigezbekia eastern herbs, roots of Scutellaria baicalensis, leaves and branches of microbiota cross-paired 57.0

while to obtain a complex extract, ethyl alcohol of 35% concentration is used, and the complex extract contains ingredients in the following ratio, kg per 100 decaliters of extract:

Green tea leaves 24.0 Turmeric roots 24.0 Seeds of Chernushka sowing 24.0 Japanese vineyard root tubers 24.0 Sigezbekia eastern grass 24.0 The roots of Scutellaria baicalensis 24.0 Leaves and branches of the microbiota cross-paired 24.0 Ethyl alcohol, 35% 280,0 Distilled water rest

Example 2. For the preparation of drugs with a cancer-preventive effect, the consumption of constituent ingredients per 100 dal, kg:

Natural honey 675.0 Green Tea Leaf Complex Extract, turmeric roots, seeds of Chernushka sowing, root of a Japanese vineyard, sigezbekia eastern herbs, roots of Scutellaria baicalensis, leaves and branches of microbiota cross-paired 60.0

at the same time, ethyl alcohol of 70% concentration is used to obtain a complex extract, and the complex extract contains ingredients in the following ratio in kg per 100 decaliters of extract:

Green tea leaves 25.0 Turmeric roots 25.0 Seeds of Chernushka sowing 25.0 Japanese vineyard root tubers 25.0 Sigezbekia eastern grass 25.0 The roots of Scutellaria baicalensis 25.0 Leaves and branches of the microbiota cross-paired 25.0 Ethyl alcohol, 35% 300,0 Distilled water rest

Example 3. For the preparation of drugs with a cancer-preventive effect, the consumption of constituent ingredients per 100 dal, kg:

Natural honey 600,0 Green Tea Leaf Complex Extract, turmeric roots, seeds of Chernushka sowing, root of a Japanese vineyard, sigezbekia eastern herbs, roots of Scutellaria baicalensis, leaves and branches of microbiota cross-paired 57.0

while to obtain a complex extract, ethyl alcohol of 35% concentration is used, and the complex extract contains ingredients in the following ratio, kg per 100 decaliters of extract:

Green tea leaves 24.0 Turmeric roots 24.0 Seeds of Chernushka sowing 24.0 Japanese vineyard root tubers 24.0 Sigezbekia eastern grass 24.0 The roots of Scutellaria baicalensis 24.0 Leaves and branches of the microbiota cross-paired 24.0 Ethyl alcohol, 70% 570.0 Distilled water rest

Example 4. For the preparation of drugs with a cancer-preventive effect, the consumption of constituent ingredients per 100 dal, kg:

Natural honey 675.0 Green Tea Leaf Complex Extract, turmeric roots Seeds of Chernushka sowing, root of a Japanese vineyard, sigezbekia eastern herbs, roots of Scutellaria baicalensis, leaves and branches of microbiota cross-paired 60.0

at the same time, ethyl alcohol of 70% concentration is used to obtain a complex extract, and the complex extract contains ingredients in the following ratio in kg per 100 decaliters of extract:

Green tea leaves 25.0 Turmeric roots 25.0 Seeds of Chernushka sowing 25.0 Japanese vineyard root tubers 25.0 Sigezbekia eastern grass 25.0 The roots of Scutellaria baicalensis 25.0 Leaves and branches of the microbiota cross-paired 25.0 Ethyl alcohol, 70% 590.0 Distilled water rest

Toxicological and pharmacological studies of the proposed drug.

Example 5. Toxicological studies

Preliminary toxicological studies on male mice of the line SD-1 and CBA (weight 22 ± 2 g) showed that the claimed drug has very low acute toxicity (LD ₅₀ > 1 g / kg) and therefore its use is safe in a wide range of concentrations.

The study of pharmacological activity was carried out on male mice of the line SD-1 and CBA (weight 22 ± 2 g). The experiments were performed on animals that were obtained from the Pushchino laboratory animal nursery and bred in a certified vivarium of TIBOKh FEB RAS. The animals were kept in accordance with international rules, approved by order No. 267 of the Ministry of Health and Social Development of the Russian Federation dated 06/19/2003, as well as the “Guidelines for the experimental (preclinical) study of new pharmacological substances” (2005).

Example 6. The study of the cancer-preventive activity of the proposed drug

To evaluate carcinogenic activity, a method of chemical carcinogenesis, which is induced in animals by a strong carcinogen substance 7,12, dimethylbenzanthracene (DMBA), was used as an experimental model [A. Farnoush * and I.C. Mackenzie Sequential histological changes and mast cell response in skin during chemically-induced carcinogenesis // J. Oral Pathology & Medicine July 1983; V 12, 14, p. 300-306].

Studies were performed on female mice of the CD-1 line. DMBA was used at a dose of 10 μg per mouse. DMBA was dissolved in benzene and applied 3 times a week for 8 weeks in a volume of 0.05 ml per shaved interscapular region of experimental animals. The studied drugs were administered orally at a dose of 10 mg / kg in a volume of 0.2 ml simultaneously with the application of a carcinogen. Animals were divided into the following groups of 10-12 animals each: 1) intact; 2) carcinogenic (negative control) - control (-); 3) carcinogenic + plant extract, designated "G35"; 4) carcinogenic + plant extract, designated "G70"; 5) carcinogenic + rosmarinic acid (positive control) - RK. As a comparison drug, we used rosmarinic acid (Sigma, USA), which has a pronounced anticarcinogenic, antioxidant, and apoptotic effect in skin carcinogenesis induced by DMBA.

After the appearance of the first papillomas, the following parameters were evaluated weekly: the number of animals with a tumor, the number of papillomas in each group, the size of the papillomas and the latent period (the period between the appearance of the first signs of tumor development until 50% of animals with a tumor in the group appeared), which was calculated by the formula:

- где F - количество опухолей, появившихся за неделю, X - количество недель, на момент учета, N - суммарное количество опухолей.

- where F is the number of tumors that appeared during the week, X is the number of weeks at the time of recording, N is the total number of tumors.

At the 15th week of the experiment, tissue samples were taken for histological and biochemical studies and blood samples were taken from experimental animals.

Example 7. Morphological evaluation of the cancer-preventive activity of the claimed drug

In experimental animals, part of the skin which was treated with DMBA, the first tumor lesions were recorded at the 7th week of the experiment in the control group (-), in 30% of the animals in the group treated with RK, G35 and G70 in 20%. Within 9-14 weeks of the experiment, marked progress in the development of the tumor process in all experimental groups was noted. By week 8, the number of mice with a tumor reached 50% in the control (-) group, in the group treated with the drug with extract G 70 at week 9, and G35 at week 10 of the experiment.

Assessment data for morphological parameters are presented in the table and in FIG. one.

Statistical and graphical processing of experimental data was carried out using the statistical package Microsoft Excel. The significance of differences was determined using the t-student parametric criterion.

FIG. 1. Parameters for changing the number of animals with tumors (a) and the sizes of these tumors (b) in different experimental groups treated with DMBA (5 weeks after the first application of the carcinogen).

Within 13 weeks, in the control (-) and G35 groups, 100% tumor formation was observed, and in the G70 treated group the number of mice with the tumor reached 90%. By 13 weeks, the tumor size in the control (-) group averaged 4.4 mm; in the G35 group, a 1.5-fold decrease in tumor size was recorded compared to the control group.

Thus, the claimed tool helps to reduce the incidence of new tumor foci, the growth rate, number and size of tumors in animals treated with DMBA. In general, the clinical picture of the manifestation of oncological pathology improves, the latent period of tumor growth significantly increases.

Example 8. Biochemical evaluation of the cancer-preventive activity of the claimed drug

To assess the development of the inflammatory process in each group, we analyzed the production of pro- and anti-inflammatory cytokines in blood serum and skin homogenate, using the Sandwich-type immunoassay enzyme-linked immunosorbent assay using the BD Bioscience OptEIA US diagnostic kits. To determine the concentration of total bilirubin, the Novogluk-K, M diagnostic kit was used (Vector-Best, Russia). The state of the antioxidant defense system in experimental animals was determined by the accumulation of secondary products of lipid pericase oxidation (lipid peroxidation) reacting with thiobarbituric acid (TBA-reactive products), the amount of which was expressed as the concentration of MDA per unit weight of protein [Karpishchenko A.I. Medical laboratory technology. - St. Petersburg: Inter-Medica, 1999].

It is known that tumor growth leads to inhibition of the immune system, which complicates the antitumor and immune response from the protective systems of the body. A comparative analysis of the level of cytokines showed that in the control (-) group, the level of both pro- and anti-inflammatory cytokines in the homogenate of the carcinogen-affected skin and slightly in the blood serum of mice of tumor carriers compared to intact animals is significantly reduced, as shown in FIG. 2.

FIG. 2. Determination of the level of pro- and anti-inflammatory cytokines in blood plasma and skin homogenate in experimental animals: IL-1 - interleukin-1; EL-17 - interleukin-17; INFy is interferon-gamma; GM-CSF - granular macrophage colony stimulating factor; TNF - tumor necrosis factor; and anti-inflammatory cytokines: IL-4 - interleukin-4; IL-10 - interleukin-10.

The data obtained show that the claimed tool helps to normalize the level of cytokines both systemically (in blood serum) and locally (in skin homogenate) to a greater extent than in the group of animals that underwent treatment of RK. The use of G35 extract stimulates the production of all studied cytokines in the skin homogenate. Moreover, G70 extract is able to increase the level of only certain cytokines in these homogenates, namely: IL-1, IL-4, and TNF. It is important to emphasize that these drugs have a similar, but less noticeable effect on the level of cytokines in blood serum.

The results show that the claimed tool is able to have a significant modulating effect on the immunological processes in the body in such a way that it stimulates its oncoprotective properties.

It is known that lipid peroxidation often occurs as a result of the development of oxidative stress at all stages of the oncological process. Analysis of the data on the content of malondialdehyde (MDA) in the blood serum of animals showed that in the negative control group, the level of MDA increases by about 1.5 times compared to the group of intact animals, as shown in FIG. 3.

FIG. 3. The content of malondialdehyde (MDA) in the blood serum of animals of different experimental groups.

Analysis of the determination of the level of total bilirubin in blood serum among the studied groups of animals shows that it does not depend on tumor growth (negative control) and inhibition of tumor growth (PK - positive control) in the studied range. In the groups that underwent treatment with G35 and especially G70 drugs, a significant decrease in this biochemical indicator is observed, which is shown in FIG. four.

FIG. 4. The content of total bilirubin in the blood serum of animals of different experimental groups.

The results of a study of the cancer-preventive activity of plant extracts G35 and G70 on a model of skin carcinogenesis induced by DMBA showed that their use leads to the correction of morphological and biochemical parameters towards their normalization in the development of skin carcinogenesis caused by DMBA.

Thus, on the basis of pharmacological studies, we can conclude that the claimed drug exhibits a pronounced carcinogenic and adaptive adaptive effect, increasing the nonspecific resistance of the body through its effect on the elements of the nervous, immune and endocrine systems, which is crucial in prevention and treatment oncological diseases.

This invention extends the arsenal of tools with cancer-preventive properties.

Claims (4)

A tool with a cancer-preventive effect, including components of plant origin, characterized in that as components of plant origin it contains a complex extract of green tea leaves, turmeric roots, sowing Chernushka roots, Japanese vineyard root tubers, eastern sisezbekia grass, Scutellaria baicalensis roots, leaves and branches microbiota cross-paired and additionally contains natural honey, in the following ratio of ingredients, kg per 100 decaliters: Natural honey 600.0-675.0 Green Tea Leaf Complex Extract, turmeric roots, seeds of Chernushka sowing, root of a Japanese vineyard, sigezbekia eastern herbs, roots of Scutellaria baicalensis, leaves and branches of microbiota cross-paired 57.0-60.0 in order to obtain a complex extract, ethyl alcohol of 35% or 70% concentration is used, and the complex extract contains ingredients in the following ratio, kg per 100 decaliters of extract: Green tea leaves 24.0-25.0 Turmeric roots 24.0-25.0 Seeds of Chernushka sowing 24.0-25.0 Japanese vineyard root tubers 24.0-25.0 Sigezbekia eastern grass 24.0-25.0 The roots of Scutellaria baicalensis 24.0-25.0 Leaves and branches of the microbiota cross-paired 24.0-25.0 Ethyl alcohol, 35% or 280.0-300.0 Ethyl alcohol, 70% 570.0-590.0 Distilled water rest

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