KR20150045036A - Composition comprising an extract of Cynanchi wilfordii for preventing or treating of climacteric disorder - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and a food composition, which include extract of Cynanchum wilfordii and of natural materials, thereby providing an anti-oxidant effect and other effects of preventing, improving and/or treating climacteric disorder.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for prevention or treatment of female climacteric diseases,

The present invention relates to a pharmaceutical composition and a food composition which contain Baekgoo extract and natural extract and exhibit a synergistic effect on antioxidant efficacy, prevention, improvement and / or treatment of female menopausal disease.

With the recent development of civilization and the progress of science, humans who live in modern life gradually prolong their lifespans. At the same time, due to stress, pollution and aging phenomenon in various environments, immunity is decreased and natural healing power gradually decreases and aging progresses. In particular, women are experiencing physical and emotional distress due to menopausal symptoms that start slowly after their forties.

Women's menopause is a kind of endocrine syndrome, which means a transition period in which physiological function and sexual function are diminished or lost due to a decrease in estrogen, a female hormone due to the general and gradual aging of ovarian function. Symptoms of these menopausal symptoms include vascular changes such as facial flushing, tachycardia, sweating or headache, and symptoms due to musculoskeletal changes such as myalgia, joint pain and back pain. There are also symptoms due to genitourinary changes such as urinary frequency or urinary incontinence, symptoms due to changes in the cerebral nervous system such as memory loss, depression, loss of concentration and dizziness, as well as visual acuity deterioration and changes in skin and hair In addition, osteoporosis and cardiovascular diseases caused by hormonal changes can cause diseases that are fatal to women's health.

Therefore, in order to improve the physical and mental health and quality of life of middle-aged women, it has been required to develop a therapeutic agent capable of improving the symptoms of menopausal symptoms. In order to improve these menopausal symptoms, hormone replacement therapy and non- steroidal drugs It has been developed. However, most of these drugs are known to have side effects such as headache and weight gain. Especially, in the case of estrogen replacement therapy, since the hormone is artificially administered to the body, (Swaran L., et al., Obsterrics & Gynecology , 91, 678-684, 1998).

Therefore, due to these problems, there is a growing interest in replacing estrogen therapy with a natural method of ingesting in the form of food or additives, and it is necessary to develop a new menopausal remedy that is effective in relieving symptoms of menopausal symptoms without side effects .

In addition, the excessively harmful active oxygen oxidizes unsaturated fatty acids, lipids and cholesterol in the body, and destroys cells in the body, resulting in deterioration of cell function and cell death. In particular, lipid, protein, sugar and DNA are irreversibly destroyed by active oxygen, resulting in various human diseases. Especially in women's menopausal disease, which begins before and after menopause, active oxygen promotes sickness, aging, and weakening of ovarian function. Accordingly, there is a growing demand for the development of antioxidants that inhibit harmful oxygen and are not toxic for the prevention or treatment of female menopausal diseases.

As a precedent study related to the treatment of menopausal disease, Korean Patent Laid-Open Publication No. 10-2011-0017419 and Korean Patent Publication No. 2010-0114333 disclose an extract of at least one member selected from the group consisting of lily, bee casualty, And a composition for preventing or alleviating menopausal symptoms comprising as an active ingredient, and Korean Patent Laid-Open No. 10-2010-0115840 provides a composition for preventing and ameliorating menopausal symptoms containing glycerol as an active ingredient .

Therefore, the inventors of the present invention have been studying a composition having antioxidative activity without side effects and having an effect of preventing or treating a menopausal disease, Wherein the mixed composition further comprises at least one natural extract selected from the group consisting of ginseng root, licorice root, ginger, jujube, dermis, The present invention has been accomplished by confirming that the composition exhibits a high total polyphenol content which is higher than that of a composition containing a single component and exhibits a synergistic therapeutic effect in antioxidant ability and treatment of menopausal symptoms.

The present invention is characterized by providing a natural-product-derived mixed composition exhibiting a synergistic effect for prevention, improvement and / or treatment of female menopausal disease.

It is an object of the present invention to provide a method for the production of an extract of Bacillus subtilis, comprising the steps of: (a) extracting Bacillus subtilis; and (b) extracting the bacterium Bacillus subtilis, And at least one kind of natural extract selected from the group consisting of wood,

It is a further object of the present invention to provide a method for the production of a composition comprising (a) Bacillus subtilis extract, and (b) Bacillus thuringiensis, Puerariae lanceolata, Leaf lily, Bellflower, Glycyrrhiza, Licorice, Chrysanthemum, Ginger, Jujube, Dermatophyte, And a gardenia extract. The present invention also provides a pharmaceutical composition for antioxidation.

It is a further object of the present invention to provide a method for the production of a composition comprising (a) Bacillus subtilis extract, and (b) Bacillus thuringiensis, Puerariae lanceolata, Leaf lily, Bellflower, Glycyrrhiza, Licorice, Chrysanthemum, Ginger, Jujube, Dermatophyte, And gum arabic. The present invention also provides a food for preventing or ameliorating female menopausal diseases.

It is a further object of the present invention to provide a method for the production of a composition comprising (a) Bacillus subtilis extract, and (b) Bacillus thuringiensis, Puerariae lanceolata, Leaf lily, Bellflower, Glycyrrhiza, Licorice, Chrysanthemum, Ginger, Jujube, Dermatophyte, Which comprises at least one natural extract selected from the group consisting of ginseng, ginseng, and gardenia.

In order to achieve the above-mentioned object, the present invention provides a method for producing a bamboo plant extract comprising (a) a bamboo plant extract, and (b) a bamboo plant, a rootstock, a lobule, a bellflower, a bellflower, , A ginseng extract, a ginseng extract, a ginseng extract, a ginseng extract, a ginseng extract, a ginseng extract, a ginseng extract, and a gardenia extract.

The present invention also relates to a composition comprising (a) Bacillus subtilis extract, and (b) Bacillus subtilis, Puerariae lanceolata, Leaf lily, Bellflower, Glycyrrhiza, Licorice, Chrysanthemum, Ginger, Jujube, Dermis, And at least one kind of natural extract selected from the group consisting of wood.

The present inventors have made efforts to develop a natural composition derived from natural products having an effect of an estrogen replacement agent used for hormone replacement therapy for postmenopausal women. As a result, it has been found that a mixture of extracts and natural extracts derived from Bacillus subtilis is excellent in phytoestrogenic and antioxidant Activity, showing a synergistic effect in effectively alleviating or preventing the symptoms of female first menopausal period or menopausal symptoms, thus completing the present invention.

As a specific example, the present invention can be a composition having 10 to 15 parts by weight per 100 parts by weight of the composition. When the extract is less than 10 parts by weight, phytoestrogen activity is insufficient and thus there is a limit in achieving an expected effect. When the extract is more than 15 parts by weight, the phytoestrogen and antioxidant activity are not further increased. There is a problem that the effect is not produced and the manufacturing cost is increased.

In another specific example, the natural extract may be 19-100 parts by weight, more preferably 20 parts by weight to 100 parts by weight, based on 100 parts by weight of the extract. When the natural extract is less than 19 parts by weight based on 100 parts by weight of the white water extract, it is difficult to expect a synergistic effect of phytoestrogen activity and antioxidant activity from the composition of the present invention composed of the natural extract to be mixed. When the natural extract is mixed in an amount exceeding 19 parts by weight with respect to 100 parts by weight, it is difficult to obtain an expected effect beyond a threshold value, resulting in an increase in manufacturing cost.

According to one embodiment of the present invention, the total polyphenol content of the composition of the present invention in which Baekgoo extract and natural extract are mixed is measured. As a result, it is found that the composition of the present invention has a much higher level Of polyphenol (Fig. 1). As a result of measuring antioxidant activity through a method known in the art, the synergistic antioxidant activity of the mixed composition was confirmed by confirming that the mixed composition had much better antioxidant ability than the single extract of Bacillus subtilis (FIGS. 2 and 3) 3).

In addition, the present inventors have found that the natural blended extract of the present invention can be used for prevention of female menopausal related diseases by estrogen activity by observing that the cell viability is increased by adding the natural mixed extract of the present invention to an estrogen-dependent MCF-7 breast cancer cell line (Fig. 4).

Therefore, it has been confirmed that the composition of the present invention can exert an excellent effect on the prevention and treatment of female menopausal disease through high total polyphenol content, antioxidant activity and phytoestrogenic activity, Respectively.

Furthermore, the present invention can be used to prevent, ameliorate, and treat various diseases caused by active oxygen as well as women's menopausal diseases since the natural-product-derived mixed composition exhibits a higher potency than the pure water extract alone . Therefore, the present invention can also be used as an antioxidant composition containing the natural product-derived mixed extract. The concentration of the white water extract and the natural mixed extract thereof contained in the pharmaceutical composition for antioxidation may be determined in consideration of the purpose of treatment, the condition of the patient, the period of time required, and the concentration is not limited to a specific range.

In another specific example, the present invention relates to a composition comprising 10 to 12 parts by weight of Bacillus subtilis extract, 8 to 9 parts by weight of Bombyx mori extract, 8 to 9 parts by weight of Puerariae Radix extract, 8 to 9 parts by weight of a lobule extract, 4-5 parts by weight of Platycodon grandiflorum extract, 4 to 5 parts by weight of folic acid extract, 4 to 5 parts by weight of licorice extract, 2 to 3 parts by weight of fescue extract, 2 to 3 parts by weight of ginger extract, 10 to 12 parts by weight of jujube extract, 8 to 9 parts by weight of dermis extract, 8 to 9 parts by weight of the extract of the early stage, 8 to 9 parts by weight of the extract of Seokchangpo, 4 to 5 parts by weight of Seomyoong Extract, 4 to 5 parts by weight of the extract of Maekmoongdong, 4 to 5 parts by weight of the extract of Green tea extract and 2 to 3 parts by weight of Gardenia extract Lt; / RTI >

In the present invention, female menopausal disease refers to the symptoms and diseases that appear in women through menopause as the estrogen secretion is reduced due to ovarian aging. For example, it can be used to treat a variety of symptoms including facial flushing, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urinary tract atrophy, vaginitis, cystitis, dysuria, And cardiovascular abnormalities such as, but not limited to, heart disease, hypertension, stroke, and the like.

The white water extract and the natural extract of the present invention may be extracted according to various methods used in a conventional method for producing natural extract such as juice extraction, steam extraction, hot water extraction, ultrasonic extraction, solvent extraction, or reflux cooling extraction, And may be preferably hydrothermally extracted. The hot water extraction can be performed by a conventional method using hot water. In order to increase the extraction yield, the natural products may be ground and used.

In the case of solvent extraction, water or an organic solvent may be used as a suitable solvent for extraction, and preferable examples thereof include purified water, methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethylacetate, Various solvents such as hexane and cyclohexane may be used alone or in combination.

When the composition of the present invention is used as a pharmaceutical composition, the content of Bacillus subtilis extract and the natural extract in the composition can be appropriately adjusted according to the symptoms of the disease, the progress of symptoms, the condition of the patient and the like, But is not limited to, 0.0001 to 99.9% by weight, and preferably 0.001 to 50% by weight. The content ratio is a value based on the dried amount from which the solvent is removed.

The composition may further contain an appropriate carrier, excipient and diluent conventionally used in the production of a pharmaceutical composition. The composition may be administered orally or parenterally in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, An external preparation, a suppository or a sterilized injection solution, and the like.

When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may include at least one excipient and / or lubricant. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like.

The preferred dosage of the composition varies depending on the condition and the weight of the patient, the degree of the disease, the drug form, the administration route and the period, but can be appropriately selected by those skilled in the art. For a more preferable effect, the dose of the composition of the present invention is preferably 0.1 mg / kg to 100 mg / kg per day based on the active ingredient, but is not limited thereto. The administration may be carried out once a day or divided into several doses. The compositions of the present invention may be administered to a mammal, including a human, in various ways. All modes of administration may be expected, for example, by oral, intravenous, intramuscular, subcutaneous injection, and the like. The pharmaceutical dosage form of the composition of the present invention may be used in the form of a pharmaceutically acceptable salt of the active ingredient, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable combination.

In another aspect, the present invention relates to a method for producing a mixture of (a) Bacillus subtilis extract, and (b) bamboo shoots, Puerariae radix, lobules, bellflower, licorice root, licorice root, ginger root, jujube root, And gum arabic. ≪ Desc / Clms Page number 2 >

The present invention also relates to a composition comprising (a) Bacillus subtilis extract, and (b) Bacillus subtilis, Puerariae lanceolata, Leaf lily, Bellflower, Glycyrrhiza, Licorice, Chrysanthemum, Ginger, Jujube, Dermis, And at least one kind of natural extract selected from the group consisting of wood.

The food and antioxidant food composition for preventing or ameliorating female menopausal disease of the present invention can be easily used as a raw material, additives, food additives, functional foods or beverages of foods.

"Food" means a natural or processed product containing one or more nutrients, preferably a state of being ready to be eaten through a certain degree of processing.

When the composition of the present invention is used as a food composition, it may contain an acceptable food-aid additive and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

The food and antioxidant composition for preventing or ameliorating female menopausal diseases according to the present invention may be, for example, various foods, beverages, gums, tea, vitamin complexes, functional foods, and the like. The food, beverage or food additive may be prepared by a conventional production method.

The white water and the natural product used in the present invention are as follows.

One. Baewooo Oh

There are two kinds of red and white, which is white cedar (Cynanchum wilfordii Hemsley) and Asclepiadaceae. They are also called other names such as mockery, dwarf grass, horsetail, new mockery, multiply root, and have the efficacy of nourishment, blood, blood, and exenteration.

2. Bounty

Bamboo is a generic term for bamboo, also known as), 죽), and 죽 竹. All bamboo is a typical medicine tree that has been used to heat up the past since its taste is sweet and its nature is cool. When you look at various oriental books, all parts of tall bamboo are used as medicine.

3. Changwon

It is called as 甘 葛, 葛 (, 鷄 根 root, 藿 藿 root, 葛 葛, and 黄 ((, and its root is used as a medicinal part. It removes symptoms of thirst, fever, vomiting and numbness, replenishes the 阴气 (phlegm), detoxifies (detoxifies), treats headache (headache) caused by wind chills (scent), opens sweat pores, I will. It treats the sore of the side by the wound which wounded by the scarf and the cold (envy), treats coughing and nauseous by infectious disease, communicates the stomach (stomach) and helps the digestion and decodes the drink.

4. Leaflet

It is a medicinal herb made of leaves of Perilla frutescens var. Acuta KUDO., A perennial herbaceous plant belonging to Lamiaceae. The seeds of lobules are called 蘇 子, and this is widely cultivated in various places. The active ingredient contains about 0.5% essential oil, and it contains 55% of perillaldehyde, cumic acid, perill alcohol and the like. The taste is a bit too warm and the fever is warm. The medicinal effect is not sweating due to the cold, it is used for the symptoms of chills and asthma sometimes with chills and fever. It is especially important for symptoms of colds with digestive disorders. In addition, when the digestive tract is called due to the food disorder, the digestive power is lowered and when the symptoms of the upper limb are appealed, the gas is relaxed while discharging the gas out.

5. Bellflower

It is a perennial plant belonging to the lily of the valley or its root. It is also called 랒 랒 줄 줄............................. Scientific name is Platycodon grandiflorum. Bellflower grows in a sunny place in the mountainous area of the temperate plain of the temperate region and the altitude of 1,000m above sea level. The height of the stem is 40 ~ 100cm. Roots can be eaten and grow thick and short. Leaves are long oval and pointed. In Aug.-Sept., The five-sided split flowers bloom, usually with bluish purple but with white flowers. It is called white bellflower, and the double blooming of flowers is called double bellflower. The bellflower roots contain many carbohydrates, calcium and iron, and fiber is the main component. So the taste of chewing is special, especially the young roots of 2 and 3 year old are very soft. It is caught from spring to autumn, and it eats raw, dried, and caught. It is good from autumn to early spring in terms of ingredients, but it is bitter in this time.

6. Fisicole

It is also called 眼 枳 实 (aan zisil), and the fruit is used as a medicinal part. The cold affects the skin, causing papular papules (papules), curing the itchy symptoms, removing the nervousness of the cold (heat), getting rid of the diarrhea (dysentery), and raising the flesh. It smoothes the function of the pentagon (the pentagon), encourages energy and lightens the body.

7. Licorice

Licorice is called ural liquorice and curved licorice, and it is called 老, 美 草, 蜜 甘, 蜜 草, Yeongtong, 甛 草, 草It is also called. It grows in Russia (Siberia), Iran, Afghanistan, Pakistan, China (Gansuk province, Shin Gangseong), Mongolia and grows in Korea. European licorice is distributed in Southern Europe, Central Asia and China. As a variant of European licorice, there are Russian licorice, Persian licorice, and Iran licorice, but not medicinal. Licorice crust is reddish brown or dark brown with vertical wrinkles and occasionally shrubs, buds and scales. The outer skin of the peeled licorice is pale yellow and fibrous. Licorice has a characteristic odor, flavor, harmonizes the toxicity of all medicines, and it shows good efficacy. It regulates the fever and morale of the book, makes good communication of all blood vessels, and strengthens muscles and bones.

8. Flower

It is a biennial herbaceous plant of Seokjuk and grows as seeds. It blooms from April to May, and white flowers bloom. A white flower blooming from early spring to early summer appears as if a small star is scattered on the ground. Young leaves and stalks of chrysanthemums are edible as herbs, especially in chrysanthemums. Rich in minerals such as protein, calcium, iron and nutrition. In particular, it makes the stomach strong, cleanses the blood and makes the milk come out well. It also treats many inflammations, such as gum disease, cavities, appendicitis, enteritis, and ulcers.

9. Ginger

It is also known as a sardine or a sardine, and Southeast Asia is a country of origin and cultivated as a vegetable. The rootstock grows sideways and is fleshy, lumpy, yellowish, with a pungent flavor and a fragrant smell. At each node of the rootstock, a fake stem made of a sheath is straight up to a height of 30 to 50 cm, and the leaves are arranged in two rows at the top. Leaves are slanted, bar-shaped, with narrow ends on both ends and long bottom leaves. Flowers do not bloom in Korea, but in the tropics, flower stalks are 20 ~ 25cm long and covered with sheaths. The flowers come out of the pores and are 4-6cm long. The calyx is short-barrel shaped, the end of the corolla is divided into 3 pieces, and the split piece is pointed. There is one operation and anther is yellow. The ovary is subordinate and the style is thin like a thread. Pharmacological action to promote gastric secretion, digestive power, cardiac excitement, blood circulation promotion, and bacillary action have been reported.

10. Jujube

It is a fruit of a jujube tree, and its color is red, so it is also called red mulberry. The red mulberry is a jujube of good quality if it is dried with cold dew. The pulp contains mainly sugar and contains mucus, maltose, and tartaric acid. Seeds contain Betuline, Betulic acid, and fat, and are widely used as medicines for diuretic, calmness, and dryness in one room.

11. Dermis

It is said to be called 피 (귀 귀 귀 귀 귀 귀 귀 귀 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 며 , Citrus unshiu Markovich (Citrus unshiu Markovich), or mature periplasm of relative plants. The dermis is said to be better in the long term, and the redder the better, so the name of Hongpie and Dermis is formed. The fruit is called tangerine (Tachibana), the leaf is tachibana (橘 葉), the yellow part where the inside of the fruit bark is removed is called 柑 紅 (橘), the seed is called 橘 核 (橘 核) . It has a peculiar odor, is slightly irritating, it is spicy, and its quality is warm. The dermis cleanses the gums and strengthens the function of the spleen, treating only the abdominal cavity, trimming, vomiting, nausea, indigestion, sluggishness, lingering symptoms, and thin stools. Seawater, sputum and diuretic action.

12. Cloth

It is a perennial herb, 60-90 cm high. The roots are long and long, and the hairs are thin and long. The stem stands right and produces many branches, and the leaves face each other. The taste is spicy, spicy, and it acts on the liver and nerves. It shows liver and god, strengthens muscles and communicates blood vessels.

13. Seokchangpo

Stone porcelain. It is a perennial plant with evergreen leaves, called white whiting, iris, stony grass, and guppy. The rootstock is thick and hard, has many nodes, and is attached to the same spot as a rock gap by taking out roots. Leaves grow from roots to base layers overlapping each other. Narrow strips are brittle and shiny. The tip of the leaf is pointed like a knife, the edge is plain and always smells good. Flowers blend in the shape of a round stick in the middle of the flower-like peduncle. The length of the stick is 5cm and the color is yellowish blue. Rootstock is mainly used as a medicinal herb and contains 0.5 ~ 0.8% essential oil in rootstock. It regulates respiration and smoothes blood circulation. It also helps to spread the airflow, and has effects such as dryness, humidification, and extinction.

14. Bokryeong

Poria cocos Wolf, which is parasitic on the root of pine, is a sclerot of scarlet which is almost scentless, slightly mucous, tastes sweet and flat, and the quality of it is not shifted to one side. . It is used for edema of the abdomen and the whole body, seaweed, vomiting, diarrhea and forgetfulness caused by nervous irritation, oil well, and also for heart edema.

15. McMundong

It is a perennial plant with a monocotyledonous plant lily lily. The name of the plant is derived from the appearance of the root. The roots of Maekmundong are used as medicines in oriental medicine. It grows well in shady areas, so it is planted in the shady garden of apartments and buildings. Leaves gather from short, coarse rootstocks to form an abandon, and often the root ends grow like peanuts. Stems stand straight and 20 to 50 cm high. Leaves are dark green with linear shape, length 30 ~ 50cm, butterfly 8 ~ 12mm and lower part like leaf sheath. The roots are used as medicines in oriental medicine, and they are used as anti-inflammatory, tonic, shinhae, expectorant, and cardiovascular agents.

16. Drake

The Chinese medicine name is sanae (砂 仁), and it is called 綠 砂 (Amamum villosum Loureiro var. xanthioides TL Wu et Senjen or Yangchun sand Amomum villosum Loureiro (ginger and Zingiberaceae) ripe berries or lumps of seed are used for medicinal purposes. It is used only for the stomach fullness of the stomach and for the edible stomach. It is effective for the vomiting diarrhea caused by 脾寒 (脾 寒). It is used in combination with other drugs when it is calm or not digested.

17. Gardenia

The gardener. It is also called Huangzi (chrysanthemum), Sichiza (水 梔子) and uses the fruit as a medicine. Evergreen is a small hardwood, often planted in the southern warm region to pick fruit. Leaves facing each other at each node are elongated elliptical or broad lanceolate. Leaf length is 3 ~ 10cm, both ends are pointed and shiny, there is no sawtooth on edge, and flat. Petioles appear to be very short and do not exist. Flowers bloom one at the end of the newly growing branch. Six to seven stamens are attached to the neck of the flower, and the fragrance surrounds them. Flowers bloom in white, but it turns yellow when seal is over. The fruit, about 3.5cm long, has 6-7 prominent lines and ripens in yellowish red in September. Fever, pain, hemostasis, and diuretic.

The natural products constituting the present invention have extremely few adverse effects on the human body as edible products and have shown through synergistic effects to mitigate or prevent various symptoms appearing in menopausal or menopausal women. The present invention has been completed upon confirming that the effect of an alternative estrogen used is sufficiently exhibited.

The white water extract of the present invention and the natural mixed extract thereof can exhibit antioxidative activity and safe phytoestrogenic activity to mitigate or prevent various symptoms appearing in menopausal or menopausal women of women and have very few adverse effects on the human body, Can be prevented.

Figure 1 is a graph showing the effect of the extract of Cynanchi wilfordii ) and a natural mixed extract thereof (mixed vegetable).
FIG. 2 is a graph showing the antioxidant activity of Bacillus anthracis extract and natural mixed extracts containing Bacillus subtilis extract according to the ABTS method.
FIG. 3 is a graph showing the antioxidative activity of the white water extract and the natural mixed extract including the white water extract and the DPPH method.
FIG. 4 is a graph showing the cell viability of MCF-7 cells of a natural mixed extract containing Bacillus subtilis extract.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Manufacturing example . Preparation of extract

10 kg of purified water, each of which is a natural substance, is washed with distilled water of 10 kg each, 30 L and extracted at a temperature of 100 ° C in a hot water bath for 6 hours to prepare respective extracts.

The prepared extracts were prepared from the extracts of Bacillus thuringiensis, Bacillus thuringiensis, Bacillus thuringiensis, Bacillus thuringiensis, Lobsters, Bellflower, Glycyrrhiza, Licorice, Chrysanthemum, Ginger, Jujube, Dermatophyte, A mixture of the extracts was prepared.

The mixing ratio of the mixture is shown in Table 1 below.

Raw material name Mixing ratio
(Parts by weight) Raw material name Mixing ratio
(Parts by weight) Baisho Oh (big mockery) 10.205 Jujube 10.205 True 8.162 Jin Pia 8.162 칡 (Chang) 8.162 A bridge 8.162 Small leaf 8.162 Stonewall 8.162 Bellflower 4.082 Boryung 4.082 Earthquake 4.082 Mac Moon Dong 4.082 Licorice 4.082 Rust 4.082 Star flower 2.040 Gardenia 2.040 ginger 2.040 Sum 100,000

Experimental Example  1. Determination of the total polyphenol content of the natural extract mixture

Polyphenols are collectively referred to as a form in which two or more hydrogens of a benzene ring are substituted with hydroxyl groups, and these include components having various physiological activities such as flavonoids. Because polyphenols have antioxidant properties, the higher the polyphenol content, the better the prevention, improvement and / or treatment of menopausal disease.

Thus, the present inventors prepared a mixture of Bacillus subtilis extract, Bacillus subtilis extract and other natural extracts, and measured the total polyphenol content. Galic acid was adjusted to 20, 40, 60, 80 and 100 ug / mL using distilled water and samples were adjusted to 50, 100, 150 and 200 ug / mL using distilled water. Galic acid and 0.5 mL of each sample were added with 1 mL of 2% Na ₂ CO ₃ , followed by reaction for 3 minutes. Folin-ciocalteu reagent (Sigma-aldrich) 0.05 mL was added and reacted for 3 minutes. Thereafter, the absorbance was measured at 720 nm.

As a result, it was confirmed that 100 ㎍ / mL of the mixture of Baekgoo extract and Table 1 contained polyphenols of 7.91 ㎍ / mL and 42.1 ㎍ / mL of Galic acid, respectively, It was confirmed that the polyphenol content was as high as about 6 times (Fig. 1).

Experimental Example  2. ABTS Law-based Antioxidant ability  Measure

The water bath was pre-operated at 70 占 폚, and 1.0 mM AAPH (2,2'-azobis- (2-amidinopropane) HCl, sigma-aldrich) And 2.5 mM ABTS2- (2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt, sigma-aldrich) Was dissolved in 100 mL of PBS [(27.117 mg AAPH + 137.175 mg ABTS2-) / 100 mL of PBS] and heated in a water bath for 30 minutes. After cooling at room temperature, it was filtered using PVDF 0.45 mm syringe filter. The standard was diluted to 20, 40, 60, 80, and 100 uL / mL with a 100 uL / mL Vit-C solution and the extract samples were diluted to 15 mg / mL and then diluted to the same ratio. After diluting with PBS to have an OD value of 0.65 ± 0.05 at a wavelength of 734 nm, 20 mL of the extract sample was added to 980 mL of ABTS solution, and reacted at 37 ° C. for 10 minutes and then measured at a wavelength of 734 nm.

The antioxidant capacity was then calculated using the formula below.

As a result of comparison with the representative antioxidant Vit-C, the mixture of Bacillus subtilis extract and the mixture of Table 1 was found to be Vit-C 7.91 Ug / mL and 27.1 ㎍ / mL, respectively, and it was confirmed that the antioxidant activity was enhanced in the mixed composition (FIG. 2).

Experimental Example  3. DPPH Law-based Antioxidant ability  Measure

The antioxidant activity assay using DPPH assay is an experimental method to confirm the DPPH radical scavenging ability of water-soluble substances and less polar substances in the sample. It has the ability to eliminate DPPH radicals having similar properties to reactive oxygen species The results show that the increase of active oxygen resistance by extract intake can be expected.

Thus, the present inventors measured the antioxidative activity of each sample using DPPH assay to confirm the radical scavenging ability of the mixture of Baekgoo extract and the mixture of Table 1. 3.94 mg of DPPH (1-diphenyl-2-picrylhydrazy, wako) was dissolved in 100 mL of EtOH and diluted with EtOH to make an O.D value of 0.7 at 517 nm to prepare a 50 uM DPPH solution. Extract samples and standard (Vit-C) stock solutions were prepared using 50% EtOH to 100 mg / 100 mL of Vit-C and 100 mg / 100 mL of sample. Dilution of the standard to 200,400,600,800,1000 ug / / mL < / RTI > and then diluted to the same ratio. standard and 2.9 mL of DPPH solution were mixed and reacted in the dark for 30 minutes. After that, 1 mL of the solution was added to the cuvette and absorbance was measured at 517 nm. The standard curve for the measured absorbance was obtained. For the control group, a mixture of 2.9 mL of DPPH and 0.1 mL of EtOH was used. Blank was measured by containing only EtOH.

The antioxidant capacity was then calculated using the formula below.

As a result of comparing the antioxidative capacity of the mixture of Baekgoo extract and the mixture of Table 1 to Vit-C, which is a representative antioxidant, the mixture of Baeksoo extract and 100 ㎍ / mL of Table 1 was Vit-C 3.5 Mu] g / mL and 13.1 [mu] g / mL, the antioxidant activity was enhanced in the mixed composition (Fig. 3).

In conclusion, the mixed composition provided by the present invention was found to have the ability to eliminate different kinds of radicals through ABTS analysis and DPPH analysis, which are two representative methods of measuring antioxidant ability, It is expected that it will also be effective in eradicating active oxygen species.

Experimental Example  4. Baewooo Oh  Of the mixed composition of Table 1 containing the extract MCF -7 cell For Cell Viability  Measure

The cell viability of the plant mixture extracts containing the Hwangsuo extracts on the MCF-7 cell line, an estrogen receptor positive human breast cancer cell line, was measured. The MTT assay using MCF-7 cells is an experimental method for confirming the efficacy of naturally occurring female hormones in females. Using a medium without FBS, conditions for menopausal women are prepared and a medium supplemented with FBS The effect of each sample can be grasped by making the condition of a general woman.

MCF-7 cells were cultured in 96-well plates using Dulveco's Modified Eagle Medium (DMEM) supplemented with FBS and DMEM medium not supplemented under different conditions. After culturing for 24 hours, the extract sample was treated and the extract sample was diluted with the third distilled water. After 24 hours, 3- (4,5-dimethylthiazol-2yl)

-2,5-diphenyl-2H-tetrazolium bromide (MTT, Sigma-aldrich) reagent was added and after 3 hours, the medium was removed and 100 μl of DMSO was added. After that, it was measured at 540 nm, and the wavelength of reference filter was set at 655 nm.

As a result, it was confirmed that a natural product mixed extract showed an effect of enhancing the viability of cells in a menopausal female model without FBS (FIG. 4). These results indicate that the extracts of the present invention have a high antioxidant ability and that the extracts are effective for preventing and improving the menopausal symptoms of women.

Claims (12)

White water extract; And
Wherein the natural extract comprises at least one natural extract selected from the group consisting of bamboo shoots, bamboo shoots, lobules, bellflowers, bamboo shoots, licorice leaves, ginger, jujube, dermis,
A pharmaceutical composition for preventing or treating female menopausal disease.
The method according to claim 1,
Wherein the white water extract is contained in an amount of 10 to 15 parts by weight based on 100 parts by weight of the composition.
The method according to claim 1,
Wherein the natural extract is contained in an amount of 19 to 100 parts by weight based on 100 parts by weight of the extract.
The method according to claim 1,
The composition comprises 10 to 12 parts by weight of Bacillus subtilis extract, 8 to 9 parts by weight of Bombyx mori extract, 8 to 9 parts by weight of Puerariae Radix extract, 8 to 9 parts by weight of leaf extract, 4 to 5 parts by weight of Platine Extract, 2 to 3 parts by weight of a ginger extract, 2 to 3 parts by weight of a ginger extract, 10 to 12 parts by weight of a jujube extract, 8 to 9 parts by weight of a dermis extract, 8 to 9 parts by weight of a ginseng extract, 8 to 9 parts by weight of the extract of Seokchangpo, 4 to 5 parts by weight of Seomyoong Extract, 4 to 5 parts by weight of Macnutaea extract, 4 to 5 parts by weight of the extract of Ganoderma lucidum and 2 to 3 parts by weight of Gardenia extract.
The method of claim 1, wherein the menopausal disease is selected from the group consisting of facial flushing, sweating, skin dryness, vaginal dryness, vaginal atrophy, lower urinary tract atrophy, vaginitis, cystitis, dysuria, , Myalgia, joint pain, or osteoporosis.
2. The pharmaceutical composition according to claim 1, wherein the white water extract and the natural extract are extracted using a hot water extraction method.
White water extract; And
Wherein the natural extract comprises at least one natural extract selected from the group consisting of bamboo shoots, bamboo shoots, lobules, bellflowers, bamboo shoots, licorice leaves, ginger, jujube, dermis,
A food composition for preventing or ameliorating a female menopausal disease.
8. The method of claim 7,
Wherein the white water extract is contained in an amount of 10 to 15 parts by weight based on 100 parts by weight of the composition.
8. The method of claim 7,
Wherein the natural extract is from 19 to 100 parts by weight based on 100 parts by weight of the extract.
8. The method of claim 7,
The composition comprises 10 to 12 parts by weight of Bacillus subtilis extract, 8 to 9 parts by weight of Bombyx mori extract, 8 to 9 parts by weight of Puerariae Radix extract, 8 to 9 parts by weight of leaf extract, 4 to 5 parts by weight of Platine Extract, 2 to 3 parts by weight of a ginger extract, 2 to 3 parts by weight of a ginger extract, 10 to 12 parts by weight of a jujube extract, 8 to 9 parts by weight of a dermis extract, 8 to 9 parts by weight of a ginseng extract, 8 to 9 parts by weight of the extract of Seokchangpo, 4 to 5 parts by weight of the extract, 4 to 5 parts by weight of the extract, 2 to 3 parts by weight of the extract of the ginger extract, and 4 to 5 parts by weight of the extract of the green tea extract.
8. The method of claim 7, wherein the menopausal disease is selected from the group consisting of facial flushing, sweating, skin dryness, vaginal dryness, vaginal atrophy, lower urinary tract atrophy, vaginitis, cystitis, dysuria, , Myalgia, joint pain, or osteoporosis.
8. The food composition according to claim 7, wherein the white water extract and the natural extract are extracted using a hot water extraction method.

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