KR101067028B1 - Composition for preventing and treating menopausal syndrome in women containing wild herbs extract - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of female menopausal syndrome containing wild herbs extract or fractions thereof, and more specifically, yunpan ( Disporum uniflorum ) or Humulus japonicus extracted with water, alcohol or a mixed solvent thereof By confirming that the extract or fractions thereof is a substance having estrogen activity by promoting expression of a promoter including an Estrogen Responsive Element (ERE) and promoting growth of MCF-7, a human breast tissue-derived cell line, It can be usefully used as a composition for preventing and treating menopausal syndrome.

Description

Composition containing the extracts or fractions of wild edible greens for the prevention and treatment of postmenopausal syndrome}

The present invention relates to a composition for the prevention and treatment of female menopausal syndrome containing wild herbs extract or fractions thereof, and more specifically, yunpan ( Disporum uniflorum ) or Humulus japonicus extracted with water, alcohol or a mixed solvent thereof It relates to a composition or health food for the prevention and treatment of female menopausal syndrome containing an extract or a fraction thereof as an active ingredient.

As women get older, they go through a series of sexual maturity processes, called puberty. It is the process of reaching a state in which an individual's physical structure and emotions can fulfill their original functions. After puberty, women secrete female hormones, estrogen and progesterone. It is the hypothalamus that controls the secretion of hormones and the pituitary gland secrets hormones under its direction. When the gonadotropin secreted by the pituitary gland transmits a signal, the hormones that control menstrual cycle and allow pregnancy are secreted by Folliocle Stimulating Hormone (FSH) and Luteizing Hormone (LH). This in turn acts on the ovary to secrete estrogen and progesterone. However, as we age, the function of the ovary decreases, which leads to a significant decrease in estrogen secretion, a decrease in response to follicle stimulating hormone (FSH) and luteinizing hormone (LH), and shortening of the early follicular phase, the menstrual cycle. Is further shortened, the ovulatory phase is further shortened, and progesterone production is reduced. Eventually, the follicles will not be able to react, producing estrogens. This cessation of ovulation is stopped menstruation is called menopause, menopause symptoms due to the reduction of hormones before or during menopause. Reduced secretion of estrogens affects the urinary and reproductive systems of women as well as other parts of the body. In postmenopausal women, the rapid decrease in estrogen causes symptoms such as hot flushes, sudden heartbeat changes, and sleep disorders, which are the most frequent menopausal symptoms. This lack of estrogen can lead to neurosis, irritability, poor concentration, depressed mood, anxiety, loss of confidence and sexual desire. In addition, due to the reduction of estrogen blood supply is not made smoothly and lose the elasticity and moisture of the skin and vaginal mucosa. For example, if the mucous membranes of your eyes, nose, and mouth are dry, you may feel uncomfortable or thirsty. Occasionally, the urinary tract or vagina may dry up, causing pain due to vaginal dryness and repeated urinary tract hepatitis. In addition, osteoporosis, atherosclerosis, and the like, which cause chronic diseases, and the need for treatment for postmenopausal women is gradually increasing, and the replacement of hormones for artificially administering estrogen as a method for treating the postmenopausal women. Hormone Replacement Therapy (HRT) is widely used.

There are two types of estrogens: natural and synthetic estrogens. Oral administration is the most widely used method of administering a synthetic estrogen hormone, and parenteral administration includes transdermal patches, transdermal creams, or gels and vaginal creams. However, the use of artificial synthetic hormones is effective in improving menopausal symptoms, but controversy has been heated as a claim that long-term use is likely to cause uterine cancer or breast cancer, venous thrombosis, and gallbladder disease. Therefore, as a substitute for the synthetic estrogens used in the hormone replacement therapy as described above, there is a trend to actively search for natural estrogens derived from natural products, especially plant-derived, having similar activities as synthetic estrogens and having fewer side effects. In particular, natural estrogens derived from plants are commonly referred to as phytoestrogen.

Phytoestrogen generally refers to a nonsteroidal compound derived from a plant that is similar in function and structure to female hormone estrogen and is found in the stem, root, flower or seed of a plant. Phytoestrogens are largely composed of soybean and isoflavone, clover, coumestan, and flax seed lignan. Others include Flavanones, Flavones, and Dihydrocalchones. Isoflavones exist in plants as inactive glycosides of Biochanin A and Formononetin, and are absorbed into the body and have a structure similar to estrogens by glucosidase of intestinal bacteria. It is converted into genistein and daidzein, and shows estrogen-like activity. As a representative example of phytoestrogens, genistein has been reported to have an anticancer effect of inhibiting the growth of cancer cells by inhibiting protein protein tyrosine kinase ( The Journal of Biological Chemistry ., 262 (12), 5592-5, 1987; Cancer Research., 49, 5111-7, 1989). However, there are also reports that it may cause cancer in people who are ineffective or have genetic predisposition to uterine and breast cancer. Therefore, it is necessary to develop new phytoestrogens that can supplement the side effects of genistein.

The young shoots of Disporum uniflorum and Humulus japonicus were edible (Jang Jun-geun, 2003, healthy wild grasses, Nexus Books) as wild herbs, which is very advantageous in terms of side effects and safety. More specifically, the Yunpanmul is a lily family and is distributed in Korea, China, Japan, and Sakhalin Island. Yunpannamul is usually planted for ornamental purposes, and young spring shoots are eaten as herbs. In folk medicine, it is used as a medicine for cough, planting or pulmonary tuberculosis. Hwansam vine has the effect of lowering the fever, making the urine go out, removing the blood and releasing the poison in the body. It also treats urine deficiency, malaria, diarrhea, dysentery, pulmonary tuberculosis, lung abscess, and pneumonia. The main herbaceous tree (本草綱目) is said to lubricate samcho (三招), to digest five grains, to supplement five intestines, to kill various insects in the stomach, and to control the whole area. It also eats young shoots sprouting in early spring.

Accordingly, the present inventors searched for new phytoestrogens for domestic native plants, especially wild vegetables, while extracts of the sperm uniflorum or Humulus japonicus showing activity against the estrogen response element. By discovering a composition comprising the active ingredient and confirming that they promote the growth of breast tissue-derived cells, the composition has a phytoestrogens activity can be used as a hormone replacement composition, thus used in the prevention and treatment of female menopausal syndrome The present invention has been completed by revealing that it can be.

An object of the present invention is a hormone replacement therapy for effectively preventing and treating menopausal syndrome symptoms in postmenopausal women, a composition for preventing and treating female menopausal syndrome containing wild herbs extract or a fraction thereof, which can replace synthetic estrogen. To provide.

In order to achieve the above object, the present invention provides a composition for preventing and treating female menopausal syndrome containing the extract of yunpanmul ( Disporum uniflorum ) or Humulus japonicus as an active ingredient.

The present invention also provides a composition for the prevention and treatment of female menopausal syndrome containing the fraction of the yunpanmul or hwansamsu vine extract as an active ingredient.

In another aspect, the present invention provides a health food for preventing and improving menopausal syndrome female containing yunpanmul or hwansam vine extract as an active ingredient.

In addition, the present invention provides a health food for the prevention and improvement of female menopausal syndrome containing the fraction of the yunpanmul or hwansamsu vine extract as an active ingredient.

The composition containing the yunpanmul or hwansamvine extract of the present invention or a fraction thereof as an active ingredient is carried out to prevent, alleviate or treat menopausal syndrome such as hot flashes, osteoporosis, venous thrombosis caused by estrogen deficiency in postmenopausal women. It can be usefully used as a composition for hormone replacement therapy (Hormone replacement therapy) (HRT), especially yunpannamul, hwansam vine is a safe substance with little side effects enough to eat young shoots as herbs.

1 is a graph showing the effect of inducing luciferase activity induced by Estrogen Responsive Element (ERE) by Yunpanmul extract.
Figure 2 is a graph showing the effect of inducing luciferase activity induced by the Estrogen Responsive Element (ERE) by the yunpanmul fraction.
Figure 3 is a graph showing the effect of inducing luciferase activity induced by the Estrogen Responsive Element (ERE) by the hwangi vine extract.
Figure 4 is a graph confirming the growth promoting effect of MCF-7 human breast-derived cell line by yunpanmul and hwansam vine extract by MTT test method.
5 is a graph confirming the growth-promoting effect of MCF-7, a breast-derived cell line by yunpanmul and hwansamvine extract by colony formation assay.

Hereinafter, terms used in the present invention will be described.

As used herein, the term "prevention" refers to any action that inhibits the symptoms of female menopausal syndrome by administration of a composition of the invention.

As used herein, the terms "treatment" and "improvement" refer to all actions in which female menopausal syndrome symptoms improve or benefit from administration of a composition of the present invention.

As used herein, the term "administration" means providing a subject with a composition of the present invention in any suitable manner.

As used herein, the term "individual" refers to any animal, including humans, monkeys, dogs, goats, pigs, or rats, having the female menopausal syndrome by administering the composition of the present invention.

As used herein, the term “pharmaceutically effective amount” means an amount sufficient to treat female menopausal syndrome at a reasonable benefit or risk ratio applicable to medical treatment, which means the type, severity, and drug of the female menopausal syndrome in the individual. Can be determined according to the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of excretion, the duration of treatment, the factors including the drug used simultaneously and other factors well known in the medical field.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for the prevention and treatment of female menopausal syndrome containing the extract of yunpanmul ( Disporum uniflorum ) or Humulus japonicus as an active ingredient.

The present invention also provides a composition for the prevention and treatment of female menopausal syndrome containing the organic solvent fractions of Yunpanmul or hwansamsu vine extract as an active ingredient.

The female menopausal syndrome is preferably any one selected from the group consisting of hot flashes, osteoporosis and venous thrombosis, but is not limited thereto.

Yunpanmul or hwansamvine extract of the present invention is preferably prepared in the following steps, but is not limited thereto:

1) extracting the dried yunpanmul or hwansam vine by adding an extraction solvent;

2) filtering the extract of step 1);

3) preparing the extract by concentrating the filtered extract of step 2) under reduced pressure; And

4) extracting the extract of step 3) with an additional organic solvent to prepare a fraction.

In the above method, yunpanmul or hwansam vine can be used without limitation, such as harvested, cultured or commercially available. The yunpan herbs or hwansam vines can be used both leaves, stems or roots, preferably the leaves and stems of the yunpan herbs, hwansam vines can be used above ground parts, but is not limited thereto. The yunpanmul or hwansam vine can be used at any time during the growth stage, but preferably a young shoot.

The extraction solvent is preferably a solvent selected from water, an alcohol or a mixture thereof, preferably a lower alcohol of C ₁ to C ₄ or a mixed solvent thereof, more preferably extracted with methanol or ethanol, but is not limited thereto. It doesn't happen. The amount of the extraction solvent is preferably 2 to 10 times the dry weight of yunpanmul or hwansam vine, but is not limited thereto. The extraction method may be an extraction method such as hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction, it may be preferably extracted one to five times by the ultrasonic extraction method. When the extraction temperature is preferably 10 ℃ to 100 ℃ and more preferably room temperature, but is not limited thereto. The extraction time is preferably 1 day to 7 days, more preferably 3 to 7 days, but is not limited thereto.

The method of preparing the extract is conventional extraction in the art, such as a method using an extraction device such as supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction or ultrasonic extraction method or using an adsorption resin containing XAD and HP-20. The method may be used, but is preferably warmed and extracted at reflux or at room temperature, but is not limited thereto. The extraction number is preferably 1 to 5 times, more preferably 3 times repeated extraction is not limited thereto.

The organic solvent is preferably obtained by sequentially fractionating n-hexane, ethyl acetate, n-butanol, and water, but is not limited thereto.

In one embodiment of the present invention, the dried leaf of yunpanmul or hwansam vines are ground to an appropriate size, put in an extraction container, ethanol is added, the solution is boiled and refluxed, left for a certain time and then filtered through a filter paper to obtain an ethanol extract. . Then, the extract of the yunpanmul or hwansam vine was further fractionated sequentially with n-hexane, ethyl acetate, n-butanol, water to obtain an n-hexane fraction, ethyl acetate fraction, n-butanol or water fraction.

In one embodiment of the present invention, MCF- transfected pERE-vitallogenin-Luc plasmid (pERE-vitallogenin-Luc plasmid) in order to determine whether the yunpanmul or hwangi vine extract or fraction has estrogen activity Luciferase activity was measured in 7 (human breast cancer cell lines) to confirm the degree of activation of the Estrogen Responsive Element (ERE). As a result, it was found that the yunpanmul or hwansamsu vine extract or fraction of the present invention significantly increased luciferase activity in a concentration-dependent manner, compared to the control group, so that the extract and fraction strongly activated the estrogen reaction site. (See FIGS. 1-3).

In another embodiment of the present invention in order to determine the effect of yunpanmul or hwansam vine extract or fraction on the growth of MCF-7 cells in hormone-deficient culture conditions, MTT test method and colony assay (Colongenic assay) It confirmed using. As a result, it was confirmed in both the MTT test method and the colony formation method that the yunpanmul or hwansam vine extract or fraction of the present invention significantly induced the growth of MCF-7 cells (see FIGS. 4 and 5).

This suppresses the growth of MCF-7 cells, which are human breast cancer-derived cell lines under culture conditions that are hormone-deficient, and increases the growth of MCF-7 cells by adding estrogen components under these conditions. Observing the growth of MCF-7 cells after treatment with specific components in the conditions can indirectly confirm the estrogen efficacy of the specific components.

Summarizing the above experimental results, it can be seen that the yunpanmul or hwansam vine extract, or fractions thereof have significant estrogen activity. That is, the yunpanmul or hwansamvine extract of the present invention or fractions thereof activate the estrogen reaction site, and by confirming that the growth of human breast cancer MCF-7 cell line under hormonal deficiency conditions, the yunpanmul or hwansamvine extract, or these It can be seen that the fraction of has phytoestrogens activity.

Therefore, the composition containing the yunpanmul or hwansamvine extract, or fractions thereof of the present invention showing estrogen activity may be usefully used as a composition for the prevention and treatment of symptoms of female menopausal syndrome caused by estrogen deficiency. .

The composition comprising the yunpanmul or hwansam vine extract of the present invention or a fraction thereof, preferably contains 0.1 to 50% by weight of the yunpanmul or hwansam vine extract or a fraction thereof based on the total weight of the composition, but is not limited thereto. .

The composition of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of a medicament.

The compositions according to the invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. have. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in the composition of the present invention, for example, starch, calcium carbonate, sucrose (sucrose), lactose (lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the preferred effect, the composition of the present invention is preferably administered at 0.0001 to 1 g / kg, preferably at 0.001 to 200 mg / kg. Administration may be administered once a day or may be divided several times. The dose is not intended to limit the scope of the invention in any way.

In another aspect, the present invention provides a health food for preventing and improving menopausal syndrome female containing yunpanmul or hwansam vine extract as an active ingredient.

In addition, the present invention provides a health food for the prevention and improvement of female menopausal syndrome containing a fraction of Yunpannamul or hwansamsu vine extract as an active ingredient.

The female menopausal syndrome is preferably any one selected from the group consisting of hot flashes, osteoporosis and venous thrombosis, but is not limited thereto.

Yunpanmul or hwansamvine extract of the present invention, or fractions thereof activate the estrogen response site, induce the growth of human breast cancer MCF-7 cell line under hormonal deficiency conditions, confirming that it has phytoestrogens activity by estrogen deficiency It can be seen that it can be usefully used as an active ingredient in the health food for preventing and improving the symptoms of female menopausal syndrome.

There is no particular limitation on the kind of food. Examples of foods to which the above-mentioned substances may be added include dairy products including drinks, meat, sausages, breads, biscuits, rice cakes, chocolates, candy, snacks, confectionery, pizza, ramen, other noodles, gums and ice cream, various soups, Beverages, alcoholic beverages, vitamin complexes, and the like, and include all healthy foods in the conventional sense.

The yunpanmul or hwansam vine extract of the present invention, or a fraction thereof, may be added to a food as it is or used with other foods or food ingredients, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose (for prevention or improvement). In general, the amount of the extract in the health food can be added to 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for health and hygiene or health control purposes, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

The health functional beverage composition of the present invention has no particular limitation on the other ingredients other than the above-mentioned extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the yunpanmul or hwansam vine extract of the present invention, or fractions thereof are various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, such as flavoring agents, coloring agents and neutralizing agents (cheese, chocolate, etc.) , Pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. In addition, the yunpanmul or hwansamsu vine extract of the present invention, or a fraction thereof may contain a pulp for the production of natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected from the range of 0.1 to about 20 parts by weight per 100 parts by weight of the yunpanmul or hwansamvine extract, or fractions thereof.

Hereinafter, the present invention will be described in detail by Examples, Experimental Examples and Preparation Examples.

However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples, Experimental Examples, and Preparation Examples.

Example 1 Preparation of Wild Vegetable Extract

In the case of yunpan herbs and hwansam vines were collected from Pyeongchang, Gangwon-do, they were washed and dried. The leafy stems were leaves and stems, and the ground ginseng vines were chopped above the ground and put into an extraction container and ethanol (Ethanol) was added. After leaving these at room temperature for 3 to 7 days, the filter paper was filtered with a filter paper to obtain yunpannamul extract and hwansam vine extract, respectively. Specifically, 700 mL of ethanol was added to 20 g of yunpanmul dried material, followed by extraction and concentration to obtain 2.8 g of extract. 21 g of dried ginseng vines were extracted three times with 900 mL of ethanol and concentrated to obtain 1.3 g of extract.

Example 2 Preparation of Wild Vegetable Fractions

 The yunpanmul extract prepared in Example 1 was sequentially partitioned into n-hexane, ethyl acetate, n-butanol and water, 144.9 mg in n-hexane, 149.3 mg in ethyl acetate, 88.3 mg in n-butanol, and 486.3 mg of fractions were obtained in water. In addition, the extract of the Korean Peninsula vine extract prepared in Example 1 was sequentially partitioned into n-hexane, ethyl acetate, n-butanol and water, and 140.5 mg in n-hexane, 145.4 mg in ethyl acetate, and 85.2 in n-butanol. 480.5 mg of the fraction could be obtained in mg and in water.

The extracts and fractions were dissolved in DMSO (dimethyl sulfoxide) and used in the following experiment.

Experimental Example 1 Determination of Estrogen Responsive Element Activation Effect by Extracts or Fractions of the Present Invention

The estrogen effect of the yunpanmul extract, hwansam vine extract, or fractions thereof prepared in <Example 1> and <Example 2> using MCF-7 breast cancer cell line (American Type of Cell Culture, ATCC) Measured. MCF-7 cells were seeded at a density of 1 × 10 ⁵ cells / well and incubated at 37 ° C., 5% CO ₂ for 24 hours. Then, the pERE-vitalogogenin-Luc plasmid was transfected (pERE-vitallogenin-Luc plasmid). After 24 hours of cultivation, the yunpanmul or hwansam vine extract or fraction dissolved in DMSO, and genistein isolated from soybean as a comparative group were treated at a concentration of 10 μM. At this time, only DMSO was treated as a control. After 24 hours, the cells were lysed, and then the luciferase activity was measured using a Dual-Luciferase assay system, and the activity of the control group was analyzed.

As a result, as shown in Table 1, and Figures 1 to 3, when the Yunpanmul extract, hwansam vine extract, and their fractions were treated for each concentration, the luciferase activity significantly increased compared to the control group. In addition, the luciferase activity was found to increase concentration-dependently (Table 1, and FIGS. 1 to 3).

sample Relative Luciferase Activity Control group 0.6 Genistein 9.8 Cultivated Sprout Extract 5 ug / ml / 10 ug / ml / 15 ug / ml 2.0 / 2.5 / 3.8 Yunfanmul Hexane Fraction 10 ug / ml / 20 ug / ml / 40 ug / ml 1.8 / 1.6 / 1.5 Yunpanmul Ethyl Acetate Fraction 10 ug / ml / 20 ug / ml / 40 ug / ml 1.8 / 2.4 / 3.8 Yunpanmul Butanol Fraction 10 ug / ml / 20 ug / ml / 40 ug / ml 1.2 / 1.8 / 2.0 Cultivated Sprout Water Fraction 10 ug / ml / 20 ug / ml / 40 ug / ml 1.2 / 1.8 / 1.9 Prickly Pear Extract 10 ug / ml / 20 ug / ml / 40 ug / ml 4.2 / 4.5 / 5.2 Round vine hexane fraction 10 ug / ml / 20 ug / ml / 40 ug / ml 4.0 / 4.2 / 4.8 Round Ginseng Ethyl Acetate Fraction 10 ug / ml / 20 ug / ml / 40 ug / ml 4.2 / 4.8 / 5.4 Round Ginseng Butanol Fraction 10 ug / ml / 20 ug / ml / 40 ug / ml 4.2 / 4.5 / 5.0 Round Ginseng Water Fraction 10 ug / ml / 20 ug / ml / 40 ug / ml 3.8 / 4.0 / 4.8

Experimental Example 2 Comparison of MCF-7 Cell Proliferation by Extracts or Fractions of the Present Invention Through the MTT Method

Measuring the degree of cell proliferation through MTT assay (MTT assay) in the yunpanmul extract or hwansam vine extract prepared in <Example 1> in the MCF-7 breast cancer cell line (American Type of Cell Culture, ATCC) It was. MCF-7 breast cancer cell lines were seeded at a density of 1 × 10 ³ cells / well and incubated at 37 ° C., 5% CO ₂ for 24 hours. After incubation for 24 hours, the ethanol extract or round vine extract dissolved in DMSO was treated by concentration in a medium containing 10% charcoal dextran treated FBS, and treated with estradiol as a positive control. Only DMSO was treated. Thereafter, the media containing each estrogen component was replaced at intervals of three days. Six days after the initial treatment, the degree of cell proliferation was compared by MTT assay.

As a result, it was found that Yunpan sprout extract or hwansam vine extract significantly increased cell proliferation of MCF-7 cell line (FIG. 4).

Experimental Example 3 Comparison of MCF-7 Cell Proliferation by Extracts or Fractions of the Present Invention Through a Colonogenic Assay

In the yunpanmul extract or hwansamsu vine extract prepared in <Example 1>, the cell proliferation of the MCF-7 breast cancer cell line (American Type of Cell Culture, ATCC) through colonization assay (Clonogenic assay) Measured. MCF-7 breast cancer cell lines were seeded at a density of 1 × 10 ³ cells / well and incubated at 37 ° C., 5% CO ₂ for 24 hours. In order to exclude the effect of estrogen contained in FBS, it was incubated for 48 hours in a medium containing 10% charcoal dextran treated FBS. Thereafter, the medium containing each estrogen component was replaced at intervals of 4 days. Genistein was treated as a positive control group, extracts of yunpanmul and hwansam vine were treated by concentration, and the control group was treated only with DMSO. After 12 days from the initial treatment, the formed cell colonies were stained with Coomasie Brillant Blue-R250, and the degree of colony formation was observed. Thereafter, photographs were taken using an i-Max ^TM Gel Image analysis system, and spot density was measured using Alpha View Software.

As a result, it was found that Yunpan sprout extract or hwansam vine extract significantly increased the cell proliferation of MCF-7 cell line compared to the control (Fig. 5).

Hereinafter, the preparation examples of the pharmaceutical composition including the extract of the present invention will be described, but the present invention is not intended to be limited thereto, but is intended to be described in detail.

Production Example 1 Preparation of Powder

Yunpanmul or Prunus vine Extract 500 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Preparation Example 2 Preparation of Tablet

Yunpanmul or Prunus vine Extract 500 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Production Example 3 Preparation of Capsule

Yunpanmul or ginseng vine fraction 500 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Preparation Example 4 Preparation of Injection

Yunpanmul or ginseng vine fraction 500 mg

Sterile sterilized water for injection

pH adjuster

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Production Example 5 Preparation of Liquid

Yunpanmul or hwansam vine fraction 100 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method for preparing a liquid, each component is added and dissolved in purified water, lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by adding purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Production Example 6 Preparation of Health Food

Yunpannamul or Prunus vine Extract 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

0.13 mg of vitamin

0.15 mg of vitamin B ₂

0.5 mg of vitamin B ₆

Vitamin B ₁₂ 0.2 g

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the above-mentioned vitamin and mineral mixture is mixed with a composition suitable for a health food in a preferred embodiment, but the compounding ratio may be arbitrarily modified. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Preparation Example 7 Preparation of Healthy Drinks

Yunpannamul or Prunus vine Extract 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components according to the conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated and then stored in a healthy beverage Used to prepare the composition.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Preparation Example 8 Preparation of Other Health Foods

<8-1> Preparation of Drink

522 mg of honey

Chioctosanamide 5 mg

Nicotinamide 10 mg

Riboflavin Sodium Hydrochloride 3 mg

Pyridoxine hydrochloride 2 mg

Inositol 30 mg

Orthoic acid 50 mg

Yunpanmul or Prunus vine Extract 0.48 ~ 1.28 mg

200 ml of water

A beverage was prepared using the above-mentioned composition and content by a conventional method.

<8-2> Preparation of Chewing Gum

Gum base 20%

Sugar 76.36-76.76%

Yunpanmul or Prunus vine Fraction 0.24 ~ 0.64%

1% fruit flavor

Water 2%

Chewing gum was prepared using the above-mentioned composition and content by a conventional method.

<8-3> Preparation of Candy

50-60% sugar

Starch syrup 39.26 ~ 49.66%

Yunpanmul or Prunus vine Fraction 0.24 ~ 0.64%

Orange flavor 0.1%

The composition and the content of the candy were prepared using a conventional method.

<8-4> Preparation of Flour Food

0.5 to 5 parts by weight of yunpanmul or hwansam vine extract was added to 100 parts by weight of wheat flour, and bread, cake, cookies, crackers and noodles were prepared using the mixture to prepare foods for health promotion.

<8-5> Production of Dairy Products

5 to 10 parts by weight of yunpanmul or hwansam vine extract was added to 100 parts by weight of milk, and various dairy products such as butter and ice cream were prepared using the milk.

<8-6> Preparation of Wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted. Black soybeans, black sesame seeds, and perilla were also steamed and dried in a known manner, and then roasted to prepare a powder having a particle size of 60 mesh. The grains and seeds produced above and the yunpannamul or hwansam vine fraction of the present invention were prepared by combining the following ratios.

Brown Rice 30%

15% rate

Barley 20%

Perilla 7%

Black Bean 7%

Black sesame 7%

Yunpanmul or Prunus vine Fraction 3%

Ganoderma 0.5%

Foxglove 0.5%

As described above, the extracts or fractions of the yunpanna rim or hwansam vine which is the composition of the present invention show distinct estrogen activity and are relatively safer than the synthetic estrogen in menopausal women because they are derived from edible materials as wild herbs. It can be usefully used as a hormone replacement composition for prevention, alleviation and treatment.

Claims (10)

A composition for preventing and treating female menopausal syndrome, comprising the extract of yunpanmul ( Disporum uniflorum ) or Humulus japonicus , or a fraction thereof.
The method of claim 1, wherein the extract is water, C ₁ ~ C ₄ lower alcohol or a mixture thereof, characterized in that the composition for preventing and treating female menopausal syndrome.
3. The composition for preventing and treating female menopausal syndrome according to claim 2, wherein the lower alcohol is methanol or ethanol.
The method of claim 1, wherein the fraction is n-hexane fraction, ethyl acetate fraction, n-butanol fraction or water fraction obtained by sequentially dividing Yunpannamul or hwansamsu vine extract with n-hexane, ethyl acetate, n-butanol and water. A composition for preventing and treating female menopausal syndrome, characterized in that.
The method of claim 1, wherein the female menopausal syndrome is any one selected from the group consisting of hot flashes, osteoporosis, and venous thrombosis.
Health food for the prevention and improvement of female menopausal syndrome containing Disporum uniflorum or Humulus japonicus extract, or fractions thereof as an active ingredient.
The method of claim 6, wherein the extract is water, lower alcohol of C ₁ ~ C ₄ or a mixture of these, characterized in that the female food for preventing and improving menopause syndrome characterized in that the extraction.
The method of claim 6, wherein the lower alcohol is a female food for preventing and improving menopause syndrome, characterized in that the ethanol.
The method of claim 6, wherein the fraction is n-hexane fraction, ethyl acetate fraction, n-butanol fraction or water fraction obtained by sequentially dividing Yunpannamul or hwansamsu vine extract with n-hexane, ethyl acetate, n-butanol and water. Health foods for the prevention and improvement of female menopause syndrome, characterized in that.
The method of claim 6, wherein the female menopausal syndrome is any one selected from the group consisting of hot flashes, osteoporosis and venous thrombosis health food for preventing and improving female menopause syndrome.

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