KR20220103334A - A composition for the improving, preventing and treating of menopausal symptome comprising polysaccharide fraction isolated from Cynanchum wilfordii - Google Patents

Abstract

The present invention relates to a composition for improving, preventing or treating female menopausal diseases. The present invention is excellent in preventing or treating female menopausal diseases by containing the polysaccharide fraction derived from cynanchum wilfordii as an active ingredient. Therefore, the present invention can be usefully utilized as food compositions and musical compositions.

Description

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to a composition capable of improving, preventing or treating female menopausal diseases by containing a polysaccharide fraction derived from baeksuo as an active ingredient.

Estrogen has a variety of biological actions, including cell number increase and differentiation. The biological actions of estrogen exert their activity through two estrogen receptor alpha (ER-α) and estrogen receptor beta (ER-β) belonging to the nuclear receptor superfamily (Nilsson et al., 2001, Physiol. Rev., 81(4):1535-1565.). The mechanism of action of the estrogen receptor (ER) is that estrogen and intracellular estrogen receptors combine to form a dimer of receptors, and these receptors are located in the promoters of the target gene, specific estrogen receptor elements (estrogen response) elements, EREs) to regulate the expression of related genes. Since the two estrogen receptors have different tissue distribution patterns, ligand binding capacity, and transactivation sites, these differences enable different selective roles of estrogen receptor agonists and antagonists in different tissues (Kuiper et al. al., 1997, Endocrinology, 138(3):863-870.). Compounds that act on estrogen receptors are being used as targets for hormone replacement therapy in postmenopausal women and chemotherapy for genital cancer.

Among the compounds capable of binding to estrogen receptors, compounds that can act as tissue selectivity and selectively agonists or inhibitors of two estrogen receptors are called selective ER modulators (SERMs) (Paech et al. ., 1997, Science, 277:1508-1510.). Representative SERMs include raloxifene and tamoxifen.

Generally known hormone preparations can supplement estrogen, but they are accompanied by the risk of heart disease, dementia and breast cancer (Chlebowski et al., 2003, JAMA, 289, (24):3243-3253.). While ER-α is known to induce the proliferation of breast cancer cells, it has been found that ER-β can be a tumor suppressor. Various attempts have been made to derive compounds and develop them as hormone replacement therapies.

Phytoestrogens are compounds that mimic or modulate the roles of estrogen produced in the body by binding to the estrogen receptor (Setchell, 1998, Am. J. Clin. Nutr., 68:1333-1346.). These compounds are structurally diverse and include flavonoids, stilbenes and lignans. Their structural features enable them to exert estrogenic or antiestrogenic activity through binding to the estrogen receptor.

Most of phytoestrogens activate the genes of both types of estrogen receptors. If phytoestrogens that selectively activate ER-α or ER-β can be discovered, it can be developed as a hormone replacement that minimizes the cancer-causing side effects of estrogen. have. Phytoestrogens, which selectively act on both estrogen receptors of ER-α or ER-β, can be usefully used for the treatment and prevention of various menopausal syndromes.

Therefore, while the present inventors were studying a substance having selective activity only on estrogen receptor beta (ER-β), the polysaccharide fraction derived from Baeksuo showed selective activity only on estrogen receptor beta (ER-β), thus preventing female menopausal syndrome and The present invention was completed by confirming that it can be usefully used for treatment.

Republic of Korea Patent No. 1544532 Republic of Korea Patent No. 0382040

An object of the present invention is to provide a food composition that can prevent or improve female menopausal diseases by containing a polysaccharide fraction derived from baeksuo as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition capable of preventing or treating female menopausal diseases by containing the polysaccharide fraction derived from baeksuo as an active ingredient.

The food composition capable of preventing or ameliorating female menopausal diseases of the present invention for achieving the above object may contain a polysaccharide fraction derived from baeksuo as an active ingredient.

The polysaccharide fraction derived from Baeksuo may contain 55 to 67% by weight of neutral sugar, 30 to 40% by weight of uronic acid, and 3 to 10% by weight of protein relative to the total polysaccharide fraction.

The neutral polysaccharide may include arabinose, rhamnose, galactose, fucose, glucose, xylose and mannose.

The neutral polysaccharide is 80 to 90 mg/g of arabinose, 20 to 30 mg/g of rhamnose, 65 to 75 mg/g of galactose, 1 to 10 mg/g of fucose, 220 to 230 mg/g of glucose relative to the total neutral polysaccharide. g, xylose 1 to 10 mg/g and mannose 5 to 15 mg/g.

The uronic acid may include galacturonic acid and glucuronic acid.

The food composition may activate estrogen receptor β (ER-β).

The female menopausal disease includes hot flashes, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urethral atrophy, vaginitis, cystitis, dysuria, urgency, difficulty concentrating, short-term memory disorder, anxiety, irritability, memory loss, muscle pain, joint pain, or It may be osteoporosis.

The polysaccharide fraction derived from baeksuo (a) extracting baeksuo with a solvent; and (b) recovering a fraction having a molecular weight of 30 kDa or more from the solvent extract extracted with the solvent.

The solvent may be water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

In addition, the pharmaceutical composition capable of preventing or treating female menopausal disease of the present invention for achieving the above other object may contain a polysaccharide fraction derived from baeksuo as an active ingredient.

The composition for improving, preventing or treating female menopausal diseases of the present invention selectively activates estrogen receptor β (ER-β) only, thereby suppressing uterine atrophy and bone loss without changing the level of estradiol to improve and prevent female menopausal diseases Or it can be treated.

In addition, since the composition of the present invention is excellent in improving, preventing or treating female menopausal diseases, it can be usefully used as a food composition, a pharmaceutical composition, and the like.

1A is a graph of measuring uterine weights in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group), and group 4 (CWE group); 1B is a graph showing the body weights of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group) after the experiment; 1C is a graph showing estradiol levels in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); 1D is a graph showing the concentration of follicle stimulating hormone (FSH) in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); 1E is a graph showing the triglyceride (TG) concentration of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); 1F is a graph showing the cholesterol concentrations in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group).
2A is a femoral radiograph (soft X-ray analysis) of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group) mice; Fig. 2B is a graph quantifying the soft X-ray photograph of Fig. 2A; FIG. 2C is a graph showing the measurement of serum alkaline phosphatase (ALP) in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group), and group 4 (CWE group); 2D is a graph of osteocalcin measurement in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group), and group 4 (CWE group); 2E is a graph showing the TRAP activity of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); 2F is a graph showing the cell viability of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); Figure 2G is a photograph stained to observe the TRAP activity of group 3 (CWPF group) and group 4 (CWE group).
Fig. 3A is a graph in which β-estradiol as a positive control was treated in ER-α expressing cells; 3B is a graph showing β-estradiol treated in ER-β expressing cells; 3C is a graph showing CWPF and CWE were treated in ER-α expressing cells, respectively; 3D is a graph showing CWPF and CWE treated in ER-β expressing cells, respectively.
4A is a graph showing the growth of MCF7 cells treated with various concentrations of E2, control and CWPF; Figure 4B is a Western blot showing the expression of phosphorylation of Akt and ERK in MCF7 cells treated with various concentrations of E2, control and CWPF; 4C is a graph quantitatively showing mRNA levels of HER2, HER3 and PTEN in MCF7 cells treated with various concentrations of E2, control and CWPF.
5A is a graph measuring the mRNA level of ER-α in the uterus of group 1 (sham group), group 2 (OVX group) and group 3 (CWPF group); Figure 5B is a graph measuring the mRNA level of ER-β in the uterus of group 1 (sham group), group 2 (OVX group) and group 3 (CWPF group); Western blot showing the expression of phosphorylation of Akt and ERK in the uterus of group 1 (sham group), group 2 (OVX group), and group 3 (CWPF group) and a graph quantifying it.

The present invention relates to a composition capable of improving, preventing or treating female menopausal diseases by containing a polysaccharide fraction derived from baeksuo as an active ingredient.

Hereinafter, the present invention will be described in detail.

The composition capable of improving, preventing or treating women's menopausal diseases of the present invention contains a polysaccharide fraction derived from baeksuo as an active ingredient.

Cynanchum wilfordii , used as a raw material for the polysaccharide fraction derived from Baeksuo, is the tuberous root of silver gourd. . By using these effects, hepatorenal insufficiency (肝腎虛證), gastric dysphoria, fertility (遺精), yoseulsanyeon, bad luck (脾虛不運), full stomach upset (脘腹脹滿), diarrhea, white hair grow faster than your age It can be used when there is insufficient milk after childbirth.

The polysaccharide fraction derived from Baeksuo of the present invention contains 55 to 67% by weight, preferably 60 to 65% by weight of neutral sugar, relative to the total polysaccharide fraction; 30 to 40% by weight of uronic acid, preferably 30 to 35% by weight; and 3 to 10% by weight of protein, preferably 3 to 5% by weight.

The uronic acid consists of galacturonic acid and glucuronic acid; The neutral polysaccharide includes arabinose, rhamnose, galactose, fucose, glucose, xylose and mannose.

Preferably, the neutral saccharides are 80 to 90 mg/g of arabinose, 20 to 30 mg/g of rhamnose, 65 to 75 mg/g of galactose, and 1 fucose relative to the total neutral polysaccharides constituting the polysaccharide fraction derived from Baeksuo. to 10 mg/g, glucose 220 to 230 mg/g, xylose 1 to 10 mg/g, and mannose 5 to 15 mg/g polysaccharide.

The polysaccharide fraction derived from baeksuo of the present invention comprises the steps of (a) extracting baeksuo with a solvent; and (b) recovering a fraction having a molecular weight of 30 kDa or more from the solvent extract extracted with the solvent;

First, in step (a), Baeksuo is extracted with a solvent to obtain a Baeksuo solvent extract.

Baeksooo used in the present invention may be used undried or dried, and may be pulverized or powdered.

The white water cucumber and the extraction solvent are mixed in a weight ratio of 1: 20 to 50, preferably 1: 35 to 45, and extracted at 90 to 110 ° C. for 5 to 10 hours.

The extraction solvent may include water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The lower alcohol may be an aqueous solution of methanol, ethanol, butanol or propanol in an amount of 20 to 99% by volume, and preferably water for improvement, prevention or treatment of female menopausal diseases.

When the content of the extraction solvent is less than the lower limit based on the white water o, the improvement, prevention or treatment effect of female menopausal disease may be reduced, and if it exceeds the upper limit, the active ingredient may be extracted in a small amount.

In addition, when the extraction temperature and extraction time are less than the lower limit, the active ingredient of baeksuo may be extracted in a small amount, and if it exceeds the upper limit, the functionality may be reduced.

Next, in step (b), a polysaccharide extract with a molecular weight of 30 kDa or more is recovered from the polysaccharide extract by removing residues from the white sorghum extract extracted with the solvent by centrifugation, solvent fractionation, or filtration, etc. do.

The method for obtaining the fraction is not particularly limited as long as it is a method for purification based on molecular weight, but preferably ultrafiltration, solvent fractionation or gel filtration chromatography, and more preferably ultrafiltration. In addition, the form of the final polysaccharide fraction may be an extract, a concentrate or a powder.

Specifically, in the present invention, a polysaccharide fraction (CWPF) derived from Baeksuo was obtained by removing the fraction having a molecular weight of less than 30 kDa using the same method as described above.

In addition, a polysaccharide purification step of removing low molecular weight substances and impurities by adding 50 to 100% alcohol having 1 to 4 carbon atoms to the final polysaccharide fraction may be added.

Unlike conventional compositions that can improve, prevent or treat female menopausal diseases, the polysaccharide fraction derived from Baeksuo prepared in this way activates estrogen receptor β (ER-β), thereby causing hot flashes, sweating, dry skin, vaginal dryness, vaginal dryness, and vaginal dryness. It can improve, prevent or treat female menopausal diseases such as atrophy, lower urethral atrophy, vaginitis, cystitis, dysuria, urgency, difficulty concentrating, short-term memory disorder, anxiety, irritability, memory loss, muscle pain, arthralgia or osteoporosis.

The composition containing the polysaccharide fraction derived from baeksuo as an active ingredient may be a pharmaceutical composition capable of preventing or treating female menopausal disease, or a food composition capable of preventing or ameliorating female menopausal disease.

As used herein, the term 'fraction' used while referring to Baeksuo includes not only a fraction obtained by treatment with an extraction solvent, but also a processed product of a polysaccharide fraction derived from Baeksuo. For example, the polysaccharide fraction derived from baeksuo may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze-drying or spray-drying.

In addition, the polysaccharide fraction derived from baeksuo of the present invention is broadly meant to include processed baeksoooo formulated to be administered to animals, for example, baeksuo powder. Although the present invention carried out the experiment with the polysaccharide fraction derived from baeksuo, it will be expected by those skilled in the art that the desired effect can be achieved even in the same form as the processed product of baeksuo.

Meanwhile, in the present specification, the term 'contained as an active ingredient' means including an amount sufficient to achieve the efficacy or activity of the polysaccharide fraction derived from baeksuo. For example, the polysaccharide fraction derived from Baeksuo is used at a concentration of 1 to 20 mg/Kg, preferably 3 to 10 mg/kg. Since the polysaccharide fraction derived from Baeksoooo is a natural product and there is no side effect to the human body even when administered in an excessive amount, the upper limit of the quantity of the polysaccharide fraction derived from Baeksuo contained in the composition of the present invention can be selected and implemented by those skilled in the art within an appropriate range.

The pharmaceutical composition of the present invention may be prepared using pharmaceutically suitable and physiologically acceptable adjuvants in addition to the active ingredients, and the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, and lubricants. agent or flavoring agent, etc. may be used.

The pharmaceutical composition may be preferably formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredients described above for administration.

Formulations of the pharmaceutical composition may be granules, powders, tablets, external preparations for skin, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectables. For example, for formulation in the form of tablets or capsules (oral preparations), the active ingredient may be combined with an orally, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or required, suitable binders, lubricants, disintegrants and color-developers may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

In the composition formulated as a liquid solution, acceptable pharmaceutical carriers are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets.

Furthermore, it can be preferably formulated according to each disease or component using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA by an appropriate method in the art.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration.

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, administration mode, age, weight, sex, morbidity, food, administration time, administration route, excretion rate, and response sensitivity of the patient, usually Thus, a skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dose of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention may be prepared in a unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient, or may be prepared by internalizing in a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.

In addition, the present invention provides a food composition for preventing or improving female menopausal diseases containing a polysaccharide fraction derived from baeksuo as an active ingredient.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, sweets, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gums, ice cream, vitamin complexes, health supplements etc.

The food composition of the present invention may include not only the polysaccharide fraction derived from baeksuo as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. include Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared as a drink or beverage, citric acid, fructose liquid, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts, etc. may be additionally included in addition to the polysaccharide fraction derived from white sorghum of the present invention. can

The present invention provides a health functional food containing a food composition comprising the polysaccharide fraction derived from Baeksuo as an active ingredient. A health functional food is a food prepared by adding a polysaccharide fraction derived from baeksuo to food materials such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspension, etc. It means to bring it, but unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long period of time using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be ingested on a daily basis. The amount of added polysaccharide fraction derived from baeksuo in such a health functional food varies depending on the type of health functional food and cannot be uniformly defined. It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of a health functional food in the form of pills, granules, tablets or capsules, it is usually added in an amount of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In addition, the present invention provides the use of a polysaccharide fraction derived from baeksuo for the manufacture of a medicament or food for the improvement, prevention or treatment of female menopausal diseases. As described above, the polysaccharide fraction derived from baeksuo may be used for improvement, prevention or treatment of female menopausal diseases.

In addition, the present invention provides a method for improving, preventing or treating female menopausal disease, comprising administering to a mammal an effective amount of a polysaccharide fraction derived from baeksuo.

As used herein, the term 'mammal' refers to a mammal that is the subject of treatment, observation or experiment, and preferably refers to a human.

As used herein, the term "effective amount" refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human as conceived by a researcher, veterinarian, physician or other clinician, which an amount that induces amelioration of the symptoms of a disease or disorder. The effective amount and frequency of administration for the active ingredient of the present invention may vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of active ingredients and other components contained in the composition, the type of formulation, and the age, weight, and general health of the patient It can be adjusted according to various factors including condition, sex and diet, administration time, administration route and secretion rate of the composition, treatment period, and concurrently used drugs. In the prevention, treatment or improvement method of the present invention, it is preferable to administer the composition at a dose of 0.001 g/kg to 10 g/kg when the composition is administered once to several times a day for adults.

In the treatment method of the present invention, the composition comprising the polysaccharide fraction derived from baeksuo as an active ingredient is administered through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a phosphorus manner.

Hereinafter, preferred examples are presented to help the understanding of the present invention, but the following examples are merely illustrative of the present invention, and it will be apparent to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It goes without saying that such variations and modifications fall within the scope of the appended claims.

Example 1. Baeksuo-derived polysaccharide fraction (CWPF)

Baeksuo grown for two years in Jeongseon, Gangwon-do was purchased (Yeongdong Herbal Agriculture Association, Jecheon, Chungcheongbuk-do), powdered and used. After mixing 44 L of water with 1 kg of white sorghum powder, extraction was performed at 95 ° C. for 6 hours, and then centrifuged at 4 ° C. at 6,500 Xg for 15 minutes to remove residues, and filtered through ultrafiltration to molecular weight. This polymer fraction of 30 kDa or more was recovered to obtain a polysaccharide fraction from white succulents.

Comparative Example 1. Baeksuo extract (CWE)

Baeksuo grown for two years in Jeongseon, Gangwon-do was purchased (Yeongdong Herbal Agriculture Association, Jecheon, Chungcheongbuk-do), powdered and used. 1 kg of Baeksuo powder was mixed with 44L of water and extracted at 95°C for 6 hours to obtain a Baeksuo hot water extract.

<Test Example>

Test Example 1. Analysis of constituent sugars of fractions

Total sugar content was determined by the phenol-sulfuric acid method using glucose as standard; Uronic acid and protein content were measured by the mhydroxydiphenyl sulfate method using galacturonic acid as a standard and the Bradford method using bovine serum albumin (BSA, SigmaAldrich, St. Louis, MO, USA), respectively.

Monosaccharide composition was determined by high performance anion exchange chromatography combined with pulsed amperometric (HPAEC-PAD).

Uronic acid in Table 1 below refers to galacturonic acid and glucuronic acid. The sugars constituting the neutral sugars indicate the content of the ingredients based on the total neutral polysaccharides.

Chemical composition (unit: wt%) Example 1 (CWPF) Comparative Example 1 (CWE) Neutral sugar 62.69±1.55 70.07±2.13 Uronic acid 32.18±2.96 27.69±1.81 Protein 4.04±0.14 1.49±0.46 Component of neutral sugar (unit: mg/g) arabinose 85.06±3.08 16.11±0.26 galactose 69.66±2.07 15.74±0.44 rhamnose 22.20±0.37 14.08±0.67 xylose 2.36±0.08 3.45±0.34 glucose 226.74±8.50 171.08±2.89 mannose 8.69±0.34 3.93±0.19 fucose 4.93±0.94 5.01±0.85 fructose 0.0 0.0 Component of Uronic acid (unit: mg/g) Galacturonic acid 167.24±3.46 27.63±1.25 Glucuronic acid 4.56±0.33 1.00±0.19

As shown in Table 1 above, it was confirmed that the crude polysaccharide fraction (CWPF) prepared according to Example 1 of the present invention had a higher content of uronic acid and protein compared to the extract of Baeksuo extract (CWE) of Comparative Example 1.

In particular, among the neutral polysaccharides, the content of arabinose, rhamnose, galactose, glucose, and mannose in the crude polysaccharide fraction (CWPF) prepared according to Example 1 was higher than that of the extract (CWE) of Comparative Example 1. high was confirmed.

Test Example 2. Animal test

laboratory animal

Ovariectomized C57BL/6J female mice (9 weeks old) were purchased from Japan SLC (Shizuoka, Japan). Mice were housed in a controlled temperature (23 °C) and 12 h light/12 h dark (12 h light, 7 am–7 am; 12 h dark, 7 pm–7 am) and normal chow (Teklab). 2018S diet, Harlan Laboratories, Indianapolis, USA) was fed. Three weeks after surgery for recovery and adaptation, the fraction of Example 1, the extract of Comparative Example 1, and vehicle (DW) were orally administered to the mice for 8 weeks (5 times per week), respectively, and body weight was measured weekly.

At the end of the experiment, the mice were anesthetized and uterine tissues were collected, and plasma samples were collected from the mice and stored at -80°C before analysis. Animal experiments were reviewed and approved by the Animal Care and Use Committee of the Korea Food Research Institute (KFRI-M-18008).

-4 groups of mice-

Group 1 (sham group): non-blanched excision control group

Group 2 (OVX group): Ovariectomy group

Group 3 (CWPF group): 200 mg/kg administration of the polysaccharide fraction of Example 1 after ovariectomy

Group 4 (CWE group): 200 mg/kg administration of the extract of Comparative Example 1 after ovariectomy

Statistical analysis is presented as mean ± SEM and Dunnett's test for post hoc analysis followed by ANOVA was used for multiple statistical comparisons. A probability value of p < 0.05 was used as the criterion for statistical significance. Data were analyzed by Student's t-test to compare the chemical composition of CWE and CWPF, and all analyzes were performed using Sigmaplot V.13.0 (Systat Software Inc., Point Richmond, CA, USA).

2-1. Determination of mouse body weight and uterine weight

1A is a graph of measuring uterine weights in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group), and group 4 (CWE group); FIG. 1B is a graph showing body weights of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group) after the experiment.

In addition, Table 2 below shows the weights of mice in each group for each time period.

Category (Unit: g) sham county OVX Army CWEF Army CWE Army before surgery 18.9±0.3 18.4±0.3 18.1±0.2 18.2±0.2 0 weeks after surgery 18.9±0.5 21.2±0.4 21.5±0.3 21.6±0.4 4 weeks after surgery 20.9±0.4 23.5±0.4 22.1±0.4 22.1±0.4 6 weeks after surgery 21.7±0.5 25.0±0.6 23.1±0.5 22.9±0.6 8 weeks after surgery 22.2±0.28 26.9±0.9 24.4±0.5 24.1±0.6

As shown in Figure 1A, at the end of the experiment to confirm the success of ovariectomy, when the uterus weight was compared between the Sham group and the OVX group, severe uterine atrophy was observed in the OVX group compared to the Sham group, It was confirmed that the uterine weight of the CWPF group and the CWE group increased by 4.1 and 2.4 times, respectively, compared to the OVX group.

Also, as shown in Table 2 and Figure 1B above, it was confirmed that the average body weight of the OVX group increased significantly compared to the Sham group after 3 weeks (week 0) after surgery ([Table 2]), and in the 8-week experiment The OVX group showed significant weight gain compared to the Sham group, but this was alleviated by administration of CWE and CWPF.

It was confirmed that the % uterus/weight ratio of each group was significantly increased in the CWPF group compared to the OVX group ( FIG. 1B ).

2-2. Determination of concentrations of estradiol, follicle stimulating hormone (FSH), triglycerides (TG) and plasma cholesterol

1C is a graph showing estradiol levels in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); 1D is a graph showing the concentration of follicle stimulating hormone (FSH) in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); 1E is a graph showing the triglyceride (TG) concentration of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); 1F is a graph showing the cholesterol concentrations in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group).

The test was measured using plasma samples from mice.

The estradiol and follicle stimulating hormone (FSH) were measured using an ELISA kit; The total triglyceride (TG) and total cholesterol (TC) were measured using a serum triglyceride measurement kit (Sigma-Aldrich) and a cholesterol kit (Biovision Inc.), respectively; The plasma high-density lipoprotein-cholesterol (HDL-c) and low-density lipoprotein cholesterol (LDL-c) were measured using a HDL and LDL/VLDL quantitative colorimetric kit (Biovision Inc.).

1C and 1D, it was confirmed that the plasma estradiol level was higher in the Sham group than in the OVX group, CWPF group and CWE group; It was confirmed that the plasma FSH concentration increased by ovariectomy (OVX group) decreased to normal level (Sham group) by administration of CWPF and CWE.

In addition, as shown in FIGS. 1E and 1F , it was confirmed that the CWPF group reduced OVX-induced hyperlipidemia by decreasing the TG, TC, HDL-c and LDL-c concentrations.

According to the prior art, estrogen treatment after oophorectomy induces uterine hypertrophy, increases plasma estradiol, and decreases plasma FSH, TG and TC in C57BL/6 mice. However, excessive estrogen treatment is limited in treatment because it can cause hormone disturbance and various gynecological diseases including breast cancer.

However, the results of the present invention confirmed that, unlike the prior art, the CWPF group exhibited estrogen-like activity in the uterus and serum lipid profile without changes in plasma estrogen levels.

2-3. Measurement of effects on bone loss and osteoclast differentiation

2A is a femoral radiograph (soft X-ray analysis) of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group) mice; Fig. 2B is a graph of quantification of the soft X-ray photograph of Fig. 2A using Image J software; FIG. 2C is a graph showing the measurement of serum alkaline phosphatase (ALP) in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group), and group 4 (CWE group); 2D is a graph of osteocalcin measurement in group 1 (sham group), group 2 (OVX group), group 3 (CWPF group), and group 4 (CWE group); 2E is a graph showing the TRAP activity of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); 2F is a graph showing the cell viability of group 1 (sham group), group 2 (OVX group), group 3 (CWPF group) and group 4 (CWE group); Figure 2G is a photograph stained to observe the TRAP activity of group 3 (CWPF group) and group 4 (CWE group) (bar: 100 μm)

At the end of the experiment, femurs were collected from the mice and used for soft X-ray analysis. Soft X-ray images were taken using a soft X-ray apparatus (DX80; SOFTEX, Tokyo, Japan) at the distal femur, and X-ray parameters were 40 kV and 40 microA for all samples. In addition, radiographs of the distal femur were quantified using ImageJ software for a relative assessment of trabecular mineral density.

In addition, alkaline phosphatase (ALP) and osteradiol (OC) were measured using an ELISA kit.

In addition, to measure TRAP activity, mouse bone marrow-derived macrophages (BMM) were prepared and α-MEM (Thermo Fisher Scientific, 10% FBS (Thermo Fisher Scientific) and macrophage colony-stimulating factor (M-CSF)) containing ockford, IL, USA). Cells were treated with or without CWE and CWPF (5-40 μg/mL each) in the presence of M-CSF (60 ng/mL) for 3 days. The effect of CWE and CWPF on cell viability was determined using the Cell Counting Kit-8 (CCK-8) assay kit (Dojindo Molecular Technologies Inc., Tokyo, Japan).

For osteoclast differentiation, BMMs were seeded at 1X10 cells/well in 6-well plates or 1X10 cells/well in 96-well plates and at 60 ng/mL M-CSF and 100 ng/well of nuclear factor-κB ligand (RANKL). mL receptor activator. CWE and CWPF (5-40 μg/mL each) were administered to BMM and cultured for 4 days. Tartrate-resistant phosphatase (TRAP) staining was performed to measure osteoclast formation. TRAP-positive multinucleated cells were observed under a microscope and presumed using the colorimetric phosphatase substrate p-nitrophenylphosphate (p-NPP).

As a result of confirming the effects of CWE and CWPF on postmenopausal osteoporosis, as shown in FIGS. 2A and 2B , the CWPF group showed significant inhibition of bone loss, but it was confirmed that the CWE group was not inhibited compared to the OVX group.

In addition, in order to evaluate the anti-osteoporosis effect of CWPF, the effect of CWPF on changes in bone turnover markers, ALP and OC, which are used as early diagnostic markers for postmenopausal osteoporosis, was investigated. As shown in FIGS. 2C and 2D, ALP and It was confirmed that the concentration of OC was decreased by the administration of CWPF, and it was confirmed that it was increased in the OVX group compared to the Sham group.

In addition, as a result of estimating the effect of CWE and CWPF on osteoclasts by measuring the osteoclast differentiation inhibitory ability using TRAP assay, the increasing CWPF concentration as shown in FIG. 2E was dosed in BMM treated with M-CSF and RANKL. It was confirmed that it exhibits a dependent TRAP activity inhibition, which means that the CWPF treatment inhibits the maturation of the differentiated osteoclasts. It was confirmed that CWE had a lower inhibitory effect on TRAP activity than CWPF.

As shown in Figure 2F, CWPF had a cell viability of 72.1% at 40 μg/mL, suggesting that this low cell viability may be responsible for the inhibitory effect of CWPF on osteoclasts.

That is, CWPF administration of Example 1 suppressed bone loss in ovariectomized mice.

2-4. Characterization as an ER-β agonist

Determining whether CWPF may affect ER signaling

ER-α and ER-β agonist assays are performed using β-estradiol, CWE and CWPF.

Fig. 3A is a graph in which β-estradiol as a positive control was treated in ER-α expressing cells; 3B is a graph showing β-estradiol treated in ER-β expressing cells; 3C is a graph showing CWPF and CWE were treated in ER-α expressing cells, respectively; 3D is a graph showing CWPF and CWE were treated in ER-β expressing cells, respectively.

A commercial human ER reporter assay panel was used to measure the reporter gene activity of estrogen receptor (ER) α and β (Cayman, Ann Arbor, MI, USA). 17β-estradiol (E2) purchased from Sigma-Aldrich was used as a positive control, and the agonist mode assay tested various concentrations of CWE and CWPF ranging from 0 to 500 μg/mL.

Briefly, ER-α and ER-β reporter cells are seeded in 96-well plates for luminescence and incubated with CWE, CWPF and E2 for 24 h. After removing the medium contents from each well, 100 μl of a luciferase detection reagent was added and incubated for 5 minutes at room temperature. Luminescence was quantified using a microplate reader (Varioskan Flash, Thermo Fisher Scientific, luminescence mode) with a read time of 0.5 s (500 ms). Relative Light Unit (RLU) was calculated as normalized expression based on maximal activity of positive agonist E2 and mean and standard deviation (n = 3) of each sample concentration was determined.

3A to 3D, it was confirmed that E2 induces both ER-α and -β expression of the luciferase reporter in a robust and reproducible manner ( FIGS. 3A and 3B ).

In addition, it was confirmed that only ER-β expression of the luciferase reporter reacted in a dose-dependent manner to CWE and CWPF treatment, and in particular, it was confirmed that ER-β expression was strongly induced with respect to CWPF treatment ( FIG. 3D ).

On the other hand, it was confirmed that ER-α was not dose-dependently affected by CWE and CWPF treatment ( FIG. 3C ).

These results suggest that CWPF may be an ER-β selective agonist, and in particular, show that CWPF has a much higher ability to activate ER-β compared to CWE at the same concentration.

Confirmation of activation of the ER-β signaling pathway but not the ER-α signaling pathway

4A is a graph showing the growth of MCF7 cells treated with various concentrations of E2, control and CWPF; Figure 4B is a Western blot showing the expression of phosphorylation of Akt and ERK in MCF7 cells treated with various concentrations of E2, control and CWPF; 4C is a graph quantitatively showing mRNA levels of HER2, HER3 and PTEN in MCF7 cells treated with various concentrations of E2, control and CWPF.

To determine whether CWPF is activated by the ER-β signaling pathway rather than the ER-α signaling pathway, the inhibitory effect on cell proliferation was tested using MCF7 cells, an ER-positive breast cancer cell line.

MCF7 human breast cancer cells were obtained from the Korean Type Culture Collection (KTCC, Seoul, Korea) and supplemented with 10% FBS (GIBCO), 100 unit/mL penicillin (Sigma-Aldrich) and 100 μg/mL streptomycin (Sigma-Aldrich). It was cultured in RPMI1640 medium (GIBCO, Grand Island, NY, USA). Cells were maintained at 37 °C in a CO2 incubator, and to determine the effect of CWPF on human breast cancer cell growth, MCF7 cells (1X 10 ⁴ cells/mL) were seeded into 96-well plates and treated with various concentrations of CWPF for 72 h. incubated during MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide; USB Corporation, Cleveland, OH, USA) solution was added to each well (final concentration: 0.5 mg/mL) 37 After removing the culture medium by incubation at ℃ for 1 hour more, dimethyl sulfoxide (Sigma-Aldrich) was applied to dissolve the formazan, and the absorbance at 570 nm (Varioskan Flash, Thermo Fisher Scientific) was detected to determine cell growth. The absorbance was measured with a multimode microplate reader (Varioskan Flash, Thermo Fisher Scientific, Waltham, MA, USA).

In addition, Western blot analysis was performed using MCF7 cells and uterine lysates. MCF7 cells were seeded in 6-well plates and fasted overnight using serum-free RPMI1640 medium. After treatment with CWPF and E2 for 6 hours, cells were harvested to obtain protein, and the protein was quantified with a BCA protein assay kit (Thermo Fisher Scientific). After blotting, polyvinylidenedifluoride membranes (Thermo Fisher Scientific) were applied to p-Akt (Ser473), Akt, p-ERK (Tyr705) and ERK (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C. It was incubated with the primary antibody. A β-actin antibody (Sigma-Aldrich) was used as a loading control, and the protein band was obtained after hybridization with a horseradish peroxidase-conjugated secondary antibody, followed by a chemiluminescence detection kit (Thermo Fisher Scientific) and Alliance Mini HD9 (UVITEC, Cambridge, UK). Quantification of band intensities was performed using ImageJ software (v1.51j).

In addition, mRNA levels in Figure 4C are determined by quantitative real-time PCR using specific primers. MCF7 cells seeded in 100 mm dishes were starved overnight with serum-free RPMI1640 medium. Cells were exposed to CWPF or E2 (10 nM) for 6 h and then harvested for RNA isolation using an RNeasy kit (Qiagen, Hilden, Germany). 1 microgram of total RNA was used for cDNA synthesis via the iScript™ cDNA synthesis kit (Biorad, Hercules, CA, USA). PCR was run on a StepOne Plus instrument (Applied Biosystems, Foster City, CA, USA) using the Fast SYBR Green Master Mix kit (Life Technologies, Gaithersburg, MD, USA). The primer sequences used in this test are presented in [Table 3]. Relative mRNA expression was determined by specific primers and normalized to TATA binding protein (TBP) for MCF7 samples and β-actin for mouse samples.

As shown in FIG. 4A , as a result of measuring cell growth of MCF7 for 3 days in the E2 treatment group, which is a positive control, significant cell proliferation was observed after 48 hours, whereas MCF7 cells treated with CWPF were treated with the control group for 48 hours. By comparison, it was confirmed that there was no significant difference. However, after treatment with CWPF of 100 μg/mL and 200 μg/mL and cultured for 72 hours, it was confirmed that the survival rate decreased by 86.2% and 84.8%, respectively.

These results suggest that CWPF acts through a distinct pathway from E2 that promotes cell proliferation.

Therefore, it is thought that CWPF acts as a selective agonist of ER-β rather than ER-α and activates ER-β-related signaling pathways.

In addition, as shown in Fig. 4B, it was observed that phosphorylation of Akt and ERK was slightly decreased in CWPF-treated MCF7 cells compared to the control vehicle-treated cells, which is consistent with the previous result of treating MCF7 cells with ER-β. do.

Also, as shown in Fig. 4C, the mRNA levels of human epidermal growth factor receptor 2 (HER2) and HER3 were reduced by CWPF treatment, and phosphatase and tensin homolog deleted from chromosome 10 (PTEN) were CWPF It was confirmed that the increase in the treated cells.

These results support that CWPF regulates ER-β signaling pathways such as PTEN/Akt and HER2/HER3 signaling.

For example, CWPF reduces the growth of MCF7 cells through activation of the ER-β signaling pathway.

Identification of the ER-β signaling pathway in the uterus

Figure 5A is a graph measuring the mRNA level of ER-α in the uterus of group 1 (sham group), group 2 (OVX group) and group 3 (CWPF group); Figure 5B is a graph measuring the mRNA level of ER-β in the uterus of group 1 (sham group), group 2 (OVX group) and group 3 (CWPF group); Western blot showing the expression of phosphorylation of Akt and ERK in the uterus of group 1 (sham group), group 2 (OVX group) and group 3 (CWPF group) and a graph quantifying it.

The phosphorylation graphs of Akt and ERK were quantified using Image J software.

As shown in FIGS. 5A and 5B , it was confirmed that the mRNA level of ER-β in the mouse uterus increased in the CWPF group compared to the OVX group, but ER-α did not.

Also, as shown in FIG. 5C , it was confirmed that phosphorylation of Akt and ERK was decreased in the uterus of CWPF-treated mice, consistent with the results of in vitro experiments using MCF7 cells.

These results suggest that CWPF alleviates oophorectomy-induced postmenopausal symptoms by regulating ER-β-related signals such as the PTEN/Akt signaling pathway.

That is, it was confirmed that the CWPF prepared according to Example 1 of the present invention specifically activated the ER-β signaling pathway, but did not induce an increase in estradiol production or ER-α activation that promotes cell proliferation.

Bone loss due to ovariectomy is caused by destruction of bone homeostasis due to an imbalance between bone formation caused by bone resorption by osteoblasts and osteoclasts. According to the prior art, it has been reported that OC and ALP levels were elevated in ovariectomized rodents compared to controls, resulting in increased bone turnover. Restoration of estrogen levels by E2 treatment protected bone loss and reduced levels of ALP and OC in an ovariectomized rodent model. Therefore, OC and ALP can be used as early diagnostic markers for postmenopausal osteoporosis.

The present invention showed that CWPF administration increased bone density in ovariectomized mice with reduced levels of ALP and OC compared to OVX mice, suggesting that CWPF reduces ovariectomized bone loss in vivo.

In addition, it was confirmed that the CWPF of the present invention inhibits TRAP activity and exhibits an inhibitory effect on osteoclast differentiation using BMM. CWPF treatment at 40 μg/mL not only reduced the cell viability of osteoclast precursors (27.9% reduction compared to the control group, Figure 2F), but also showed an osteoclast differentiation inhibitory effect (77.8% inhibition compared to the control group, Figure 2E). Although the inhibitory effect on differentiation is greater than that of osteoclasts on reducing cell viability, it is clear that the cell viability of osteoclasts can be affected by CWPF treatment.

Estrogen induces osteoclast apoptosis mediated by nonclassical ER-α action. In the present invention, it can be suggested that the effect of CWPF on osteoclast apoptosis is mediated by ER-β activation.

It can also be seen that reduced FSH levels in mice administered CWPF alleviated bone loss due to ovariectomy ( FIG. 1D ). Previously, deletion of FSH receptors in mice significantly reduced bone loss after ovariectomy, supporting the idea that FSH is mainly responsible for postmenopausal bone loss.

Therefore, it was suggested that the CWPF of the present invention may help alleviate postmenopausal osteoporosis by reducing bone loss due to loss of ovarian hormone.

Furthermore, the present invention confirmed that CWPF is a selective agonist for ER-β. It has been previously proposed that ER-β regulates the PTEN/PI3K/Akt signaling pathway by down-regulating the expression of HER2 and HER3 in breast cancer cells. The experiments of the present invention showed that the effect of CWPF on the PTEN/Akt signaling pathway (PTEN up-regulation and Akt down-regulation) was significant both in vitro and in vivo.

In MCF7 cells, estradiol rapidly induced phosphorylation of ERK, which is differently regulated by ER-α and -β ( FIG. 4B ). ERK phosphorylation changes were observed to decrease in MCF7 cells and in the uterus of CWPF-administered mice ( FIGS. 4B and 5C ).

These results suggest that CWPF treatment modulates ER-β-related signals such as PTEN/Akt and ERK signaling pathways to alleviate menopausal symptoms and prevent estrogenic effects.

Hereinafter, formulation examples of the composition containing the powder of the present invention will be described, but the present invention is not intended to limit the present invention, but merely to describe it in detail.

Formulation Example 1. Preparation of powder

500 mg of the polysaccharide fraction derived from Baeksuo obtained in Example 1

Lactose 100 mg

talc 10 mg

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

Formulation Example 2. Preparation of tablets

300 mg of the polysaccharide fraction derived from Baeksuo obtained in Example 1

100 mg cornstarch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional manufacturing method of tablets.

Formulation Example 3. Preparation of capsules

200 mg of the polysaccharide fraction derived from Baeksuo obtained in Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

0.2 mg magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled in a gelatin capsule to prepare a capsule.

Formulation Example 4. Preparation of injection

600 mg of polysaccharide fraction derived from Baeksuo obtained in Example 1

mannitol 180 mg

2974 mg of sterile distilled water for injection

Na ₂ HPO _4, 12H ₂ O 26 mg

According to a conventional method for preparing injections, the content of the above ingredients per 1 ampoule is prepared.

Formulation Example 5. Preparation of liquid formulation

4 g of the polysaccharide fraction derived from Baeksuo obtained in Example 1

10 g isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to a conventional liquid preparation method, each component is added and dissolved in purified water, an appropriate amount of lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 g by adding purified water, and then filled in a brown bottle and sterilized to prepare a liquid.

Formulation Example 6. Preparation of granules

1,000 mg of the polysaccharide fraction derived from Baeksuo obtained in Example 1

appropriate amount of vitamin mixture

70 μg vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

0.2 μg of vitamin B12

Vitamin C 10 mg

Biotin 10 μg

1.7 mg of nicotinic acid amide

50 μg of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture appropriate amount

ferrous sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

potassium phosphate monobasic 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

100 mg calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for granules in a preferred embodiment, but the mixing ratio may be arbitrarily modified. It can be prepared and used in the preparation of a health functional food composition according to a conventional method.

Formulation Example 7. Preparation of functional beverage

1,000 mg of the polysaccharide fraction derived from Baeksuo obtained in Example 1

1,000 mg citric acid

100 g of oligosaccharides

2 g of plum concentrate

1 g taurine

Add purified water to total 900 mL

After mixing the above ingredients according to the usual health drink manufacturing method, after stirring and heating at 85 ° C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, then refrigerated. It is used to prepare the functional beverage composition of the present invention.

Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demand class, demanding country, and use.

Claims (11)

A food composition for preventing or improving women's menopausal diseases, comprising a polysaccharide fraction derived from baeksuo as an active ingredient. According to claim 1, wherein the polysaccharide fraction derived from Baeksuo contains 55 to 67% by weight of neutral sugar, 30 to 40% by weight of uronic acid, and 3 to 10% by weight of protein relative to the total polysaccharide fraction. Food composition for preventing or improving female menopausal disease, characterized in that. According to claim 2, wherein the neutral polysaccharide comprises arabinose, rhamnose, galactose, fucose, glucose, xylose and mannose food composition for preventing or improving female menopausal disease. The method according to claim 2, wherein the neutral polysaccharide is 80 to 90 mg/g of arabinose, 20 to 30 mg/g of rhamnose, 65 to 75 mg/g of galactose, 1 to 10 mg/g of fucose, relative to the total neutral polysaccharide, Glucose 220 to 230 mg/g, xylose 1 to 10 mg/g and mannose 5 to 15 mg/g A food composition for preventing or improving female menopausal disease. The food composition for preventing or improving female menopausal diseases according to claim 2, wherein the uronic acid comprises galacturonic acid and glucuronic acid. The food composition for preventing or improving female menopausal disease according to claim 1, wherein the food composition activates estrogen receptor β (ER-β). The method of claim 1, wherein the female menopausal disease is hot flashes, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urethral atrophy, vaginitis, cystitis, dysuria, urgency, difficulty concentrating, short-term memory disorder, anxiety, nervousness, memory. A food composition for the prevention or improvement of female menopausal disease, characterized in that it is decline, muscle pain, arthralgia or osteoporosis. The method of claim 1, wherein the polysaccharide fraction derived from baeksuo is prepared by the steps of: (a) extracting baeksuo with a solvent; and
(b) recovering a fraction with a molecular weight of 30 kDa or more from the solvent extract extracted with the solvent; The food composition for preventing or improving female menopausal disease according to claim 8, wherein the solvent is water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. A pharmaceutical composition for the prevention or treatment of female menopausal diseases comprising a polysaccharide fraction derived from Baeksuo as an active ingredient. 11. The method of claim 10, wherein the polysaccharide fraction derived from Baeksuo contains 55 to 67% by weight of neutral sugar, 30 to 40% by weight of uronic acid, and 3 to 10% by weight of protein relative to the total polysaccharide fraction. A pharmaceutical composition for preventing or treating female menopausal disease, characterized in that.

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