KR102526718B1 - Composition for the preventing and improving menopausal disorder, comprising extract of acer tegmentosum maxim and extract of Rumecis Radix - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of menopausal diseases comprising a mixed extract or a fraction thereof of Santianmok and Yangjegeun as an active ingredient, a method for preventing or treating menopausal diseases using the pharmaceutical composition, and a mixed extract of Santianmok and Yangjegeun or a pharmaceutical composition thereof It relates to a health functional food composition for preventing or improving menopausal diseases containing fractions, and a feed composition for preventing or improving menopausal diseases containing a mixed extract or fractions of Santenmok and Yangjegeun. The composition containing the mixed extract of Santenmok and Yangjegeun provided in the present invention can effectively improve various symptoms such as reduced levels of ALP, TC, LDL and osteocalcin in the blood, symptoms of depression, and increased fat cell size in menopausal diseases. Therefore, it will be widely used in the development of therapeutic agents for menopausal diseases.

Description

Composition for the preventing and improving menopausal disorder, comprising extract of acer tegmentosum maxim and extract of Rumecis Radix}

The present invention relates to a composition for preventing and ameliorating menopausal diseases comprising an extract of Santianmok and Yangjegeun, and more specifically, the present invention relates to a composition for preventing or treating menopausal diseases comprising a mixed extract of Santianmok and Yangjegeun or a fraction thereof as an active ingredient Pharmaceutical composition, a method for preventing or treating menopausal diseases using the pharmaceutical composition, a health functional food composition for preventing or improving menopausal diseases containing a mixed extract or fractions of Santenmok and Yangjegeun, and a mixed extract or fractions thereof It relates to a feed composition for preventing or improving menopausal diseases comprising a.

Menopause is a transitional period from sexual maturity to old age, and endocrine function, especially ovarian function, declines, and menstrual irregularities and anovulation gradually reach menopause, accompanied by atrophy of the genital organs and systemic aging. There are individual differences in menopause, but it usually falls between the ages of 45 and 55. During this period, various symptoms such as paroxysmal excitement, facial flushing, headache, palpitations, dizziness, tinnitus, and vasomotor disorders such as insomnia, gastrointestinal disorders, and mental disorders are often displayed, and the syndrome accompanied by abnormal symptoms is called climacteric disorder It is said. In particular, the characteristic physical changes of menopausal women include weight gain and bone density decrease. As estrogen secretion decreases, fat cell production increases, body weight increases, and the incidence of osteoporosis increases as the female hormone estrogen decreases and bone density decreases due to calcium lost together.

Osteoporosis is a disease in which bone mass decreases due to various causes and the risk of fracture continuously increases due to degeneration of the microstructure of bone tissue. It occurs when the balance of formation is broken and the osteoclast action is increased more than the osteoblast action. Normal bones form a dense structure like a mesh, but in the case of osteoporosis, the gap between bone microstructures widens and the microstructure becomes thinner and weaker, increasing the risk of easy bone fracture even with a small impact. Postmenopausal osteoporosis It is classified into senile osteoporosis, which gradually develops in men and women over 70 years of age and causes gradual bone loss of the pelvic and vertebral bones, and secondary osteoporosis caused by diseases, drugs, alcohol, smoking, or accidents regardless of age.

Osteoporosis is currently one of the most important social problems, and in the case of the United States, it causes about 260,000 women every year, and about 12 to 20% of them lead to death. In a situation where society is aging and women's participation in society is increasing, osteoporosis and fractures due to osteoporosis in the elderly or postmenopausal women cause serious problems.

Substances currently used as osteoporosis treatments include estrogen, androgenic nagolic ateroid, calcium preparations, ginseng salts, fluoride preparations, ipriflavone, and vitamin D3. Estrogen inhibits apoptosis of osteoblast cells to increase cell survival period and promotes apoptosis of osteoclast cells to decrease cell survival period, which is a somewhat effective method for treating menopausal symptoms and maintaining bone density, breast cancer, and endometrial hyperplasia There are side effects that can cause In addition, calcitonin, parathyroid hormone, and bisphosphonate preparations are examples of drugs that suppress bone destruction by inhibiting the activity of osteoclasts or increase the activity of bone regeneration units through the proliferation of osteoblasts. However, existing osteoporosis treatment drugs only block bone resorption or have only the effect of promoting bone formation, and cause many side effects when administered for a long time. Therefore, there is a demand for the development of safe preventive and therapeutic agents that exhibit a continuous increase in bone density even during long-term administration and have fewer side effects.

Accordingly, studies are being actively conducted to develop therapeutic agents containing ingredients derived from natural products as active ingredients. A pharmaceutical composition for preventing or treating menopausal symptoms containing is disclosed.

Under this background, the present inventors have made diligent research efforts to develop a formulation that can more effectively and safely alleviate menopausal diseases. , completed the present invention.

One object of the present invention is to provide a pharmaceutical composition for preventing or treating menopausal diseases comprising a mixed extract or fractions of Santianmok and Yangjegeun.

Another object of the present invention is to provide a method for preventing or treating menopausal diseases using the pharmaceutical composition.

Another object of the present invention is to provide a health functional food composition for preventing or improving menopausal diseases containing a mixed extract or fractions of Sancheonmok and Yangjegeun.

Another object of the present invention is to provide a feed composition for preventing or improving menopausal diseases comprising a mixed extract or fractions of Santianmok and Yangjegeun.

One embodiment of the present invention for achieving the above object provides a pharmaceutical composition for preventing or treating menopausal diseases comprising a mixed extract or fractions of Santianmok and Yangjegeun.

The term "acer tegmentosum maxim" of the present invention is a plant belonging to the Maple family, which is called Sancheongmok, bee tree, and mountain axim. It is thick and the material is white and light. Since Sancheonmok is non-toxic, it is a medicine with little side effects for any constitution, and it is also used as a blood purifying agent and water purifying agent because of its light taste. It restores the temperature of the liver to normal and allows moisture to be excreted well. In Korea, the leaves and xylem are used to treat liver diseases such as hepatitis, liver cirrhosis, and liver cancer, as well as to treat leukemia, diabetes, nephritis or edema, and are used for alcohol detoxification. It is also used for this purpose. In addition, it has been reported that taking the extract of Santenmok xylem while drinking alcohol can prevent alcohol poisoning.

In the present invention, the Santenmok may be purchased and used commercially or collected or cultivated in nature, but is not limited thereto.

The term "Rumecis Radix" of the present invention refers to the root of Rumex japonicus Houttuyn. or a related plant of the same genus. Morphologically, it is a long columnar shape with horizontal wrinkles and thin branch roots, and the remains of the stem remain at the root, the outer surface is grayish yellow or yellowish brown, and the bent surface is relatively flat and yellowish brown. It is known that dark brown lines appear concentrically similar to tree rings. Yangje (羊蹄) is the name given because the shape of the root resembles a sheep's hoof, and it is known that it can be used as a raw material after lightly blanched. In oriental medicine, the properties of Yangjegeun are defined as having a peculiar smell, bitter taste, cold nature, and a little poison. The yangjegeun has the effect of turning off blood heat and hemostasis by acting on blood, so it can be used for nosebleeds, vomiting blood, fecal bleeding, hemoptysis, uterine bleeding, scabies, ringworm, constipation, etc. Specific pharmacological activities include shortening of blood coagulation time and inhibition of skin fungi , cold prevention, and leukemia suppression effects are known.

In the present invention, the yangjegeun may be purchased and used commercially, or harvested or cultivated in nature, but is not limited thereto.

As used herein, the term "extract" refers to a liquid component obtained by immersing a desired substance in various solvents and then extracting the substance at room temperature or at elevated temperature for a certain period of time, and a solid component obtained by removing the solvent from the liquid component. means In addition, in addition to the above results, it can be comprehensively interpreted as including all dilutions of the results, concentrates thereof, adjusted products, and purified products thereof.

Accordingly, the mixed extract provided in the present invention is an extract obtained by extracting a mixture of Santianmok and Yangjegeun, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, or a mixture thereof. It can be interpreted as including extracts of all formulations that can be formed using extracts themselves and extracts, such as mixtures.

In addition, the mixture of Santianmok and Yangjegeun can be interpreted as a mixture of all or parts (flowers, leaves, fruits, stems, roots, bark, sap, etc.) of Santianmok and Yangjegeun.

In addition, the mixed extract can be interpreted as including even a mixture of Santenmok extract and Yangjegeun extract.

The content ratio of Sancheonmok and Yangjegeun included in the mixed extract is not particularly limited thereto, but as an example, the content ratio of Sancheonmok and Yangjegeun may be 9:1 to 1:9 (w/w). As another example, The content ratio of Santianmok and Yangjegeun may be 7:3 to 3:7 (w/w), and as another example, the content ratio of Santianmok and Yangjegeun may be 6:4 (w/w).

In the present invention, the extract can be obtained by extracting Santianmok, Yangjegeun, or a mixture thereof with water or various organic solvents. At this time, the organic solvent used is not particularly limited thereto, but as an example, it may be water, a polar solvent or a non-polar solvent, and as another example, water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol or butanol) etc.), a mixed solvent thereof, and the like, and as another example, ethanol or a mixed solvent thereof may be used.

In addition, the method for obtaining the extract is also not particularly limited thereto, but preferably, the whole plant of Santianmok, Yangjegeun or a mixture thereof, dried product, processed product, etc. thereof is immersed in the solvent, and the room temperature is 10 to 25 ° C. Methods such as a cold extraction method extracted from, a heating extraction method extracted by heating at 40 to 100 ° C., an ultrasonic extraction method extracted by applying ultrasonic waves, and a reflux extraction method using a reflux condenser may be used.

The term "fraction" of the present invention refers to a product obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various components.

In the present invention, the fraction may be interpreted as a fraction obtained by applying the mixed extract to various fractionation methods. Fractions of the extract can be obtained by applying the extract to various fractionation methods. The fractionation methods are not particularly limited thereto, but solvent fractionation methods performed by treating various solvents and ultrafiltration membranes having a constant molecular weight cut-off value It may be an ultrafiltration fractionation method performed by passing through, a chromatographic fractionation method performing various chromatography (made for separation according to size, charge, hydrophobicity or affinity), and the like. In particular, the solvent used in the solvent fractionation method is not particularly limited thereto, but a polar solvent or a non-polar solvent may be used, and preferably a non-polar solvent may be used. The solvent fractionation method may be performed by sequentially fractionating the extract using a solvent having a low non-polar level from a solvent having a high non-polar level. For example, a method of sequentially fractionating the extract using hexane or ethyl acetate may be used. can

The term "menopausal disease" of the present invention, also referred to as climacteric disorder, refers to a disease that appears during menopause. The menopause is a transitional period from sexual maturity to old age, and endocrine function, especially ovarian function, declines, and menstrual irregularity, anovulation, etc. gradually reach menopause, accompanied by atrophy of the genital organs and systemic aging. There are individual differences in menopause, but it usually falls between the ages of 45 and 55. During this period, various symptoms such as paroxysmal excitement, facial flushing, headache, palpitations, dizziness, tinnitus, vasomotor disorder such as insomnia, gastrointestinal disorder, and mental disorder are often displayed. do.

In the present invention, the menopausal disease is not particularly limited thereto, but may include, for example, weight gain, bone disease, facial flushing, headache, gastrointestinal disorder, depression, vaginal dryness, and the like. It can be a variety of bone diseases such as hypoplasia, rheumatoid arthritis, bone cancer, and bone metastases.

The term "prevention" of the present invention refers to all actions to suppress or delay the onset of the menopausal disease by using the mixed extract of Santianmok and Yangjegeun or a pharmaceutical composition containing the same according to the present invention.

According to one embodiment of the present invention, the mixed extract of Santianmok and Yangjegeun provided in the present invention reduces body weight and weight of adipose tissue in an ovariectomized animal model designed to simulate menopausal diseases (FIG. 1a and 1b) , Reduces the levels of ALP (Alkaline phosphatase), TC (Total cholesterol) and LDL (Low density lipoprotein) whose content is increased in blood by ovariectomy (Fig. 2a to 2c), and blood content by ovariectomy It reduces the increased level of osteocalcin (Fig. 3), improves depression symptoms (Figs. 4a and 4b), and reduces the size of adipocytes increased by ovariectomization (Figs. 5a and 5b).

The pharmaceutical composition of the present invention may include 0.001 to 80, specifically 0.001 to 70, and more specifically 0.001 to 60% by weight of the mixed extract based on the total weight of the composition, but is not limited thereto.

In addition, the pharmaceutical composition may further include a pharmaceutically acceptable carrier, excipient, or diluent commonly used in the preparation of pharmaceutical compositions, and the carrier may include a non-naturally occurring carrier. there is. The carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In addition, the pharmaceutical composition may be formulated according to conventional methods such as tablets, pills, powders, granules, capsules, suspensions, internal solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and transdermal formulations. Absorbents, gels, lotions, ointments, creams, patches, cataplasma agents, pastes, sprays, skin emulsions, skin suspensions, transdermal delivery patches, drug-containing bandages, or suppositories may be formulated and used. Specifically, when formulated, it may be prepared using diluents or excipients such as commonly used fillers, weighting agents, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. Such a solid preparation may be prepared by mixing at least one or more excipients, for example, starch, calcium carbonate, sucrose, lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate and talc may also be used in addition to simple excipients. It may be prepared by adding various excipients, for example, wetting agents, sweeteners, aromatics, and preservatives, in addition to liquids and liquid paraffin for oral use. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for the suppository, Witepsol, Macrogol, Tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.

Another embodiment of the present invention provides a method for preventing or treating menopausal diseases, comprising administering the pharmaceutical composition to a non-human subject in need of prevention or treatment of menopausal diseases.

The term "administration" of the present invention means the act of introducing the pharmaceutical composition to a subject by an appropriate method.

The term "subject" of the present invention refers to all animals, such as rats, mice, livestock, including humans, for which prevention or treatment of menopausal diseases is required. As a specific example, it may be mammals including humans.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is dependent on the type and severity of the subject, age, sex, drug activity, It may be determined according to factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used concurrently, and other factors well known in the medical field. For example, the mixed extract may be administered at a dose of 0.01 to 500 mg/kg per day, specifically 10 to 100 mg/kg, and the administration may be administered once or several times a day.

The pharmaceutical composition may be administered alone or in combination with other immune enhancing agents, and may be administered sequentially or simultaneously with conventional immune enhancing agents. And it can be single or multiple administrations. It is important to administer the amount that can obtain the maximum effect with the minimum amount without side effects in consideration of all the above factors, and can be easily determined by those skilled in the art.

In addition, the pharmaceutical composition may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally or topically applied) according to the desired method, and the dosage is the patient's condition and weight, the degree of disease , Depending on the drug form, administration route and time, it can be appropriately selected by those skilled in the art.

Another embodiment of the present invention provides a health functional food composition for preventing or improving menopausal diseases comprising a mixed extract or fractions of Santianmok and Yangjegeun.

The term "improvement" of the present invention refers to all activities that improve or benefit symptoms of suspected or affected individuals of menopausal diseases by using a health functional food composition for preventing or improving menopausal diseases containing a mixed extract of Sancheonmok and Yangjegeun. say

The health functional food composition for preventing or improving menopausal diseases of the present invention includes pills, powders, granules, infusions, tablets, capsules, or liquids, and foods to which the composition of the present invention can be added include, for example, , various foods, for example, beverages, gum, tea, vitamin complexes, health supplements, and the like.

Except for the mixed extract of Santianmok and Yangjegeun as essential ingredients that can be included in the food composition of the present invention, there are no particular restrictions on other ingredients, and various herbal extracts, food supplement additives or natural carbohydrates, etc. may be included as an additional component.

In addition, the food auxiliary additives include food auxiliary additives common in the art, such as flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, and the like.

Examples of the natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. In addition to the above, natural flavors (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can advantageously be used as flavoring agents.

In addition to the above, the food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like may be contained. In addition, it may contain fruit flesh for the manufacture of natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination.

Specifically, the health supplement includes health functional food and health food. The health functional food (functional food) is the same term as food for special health use (FoSHU), and is a medicine processed to efficiently display bioregulatory functions in addition to nutritional supply, and has high medical effect. means food. Here, "function (sex)" means to obtain useful effects for health purposes such as regulating nutrients for the structure and function of the human body or physiological functions. The food of the present invention can be prepared by a method commonly used in the art, and can be prepared by adding raw materials and ingredients commonly added in the art during the preparation. In addition, the formulation of the food may also be prepared without limitation as long as the formulation is recognized as a food. The food composition of the present invention can be prepared in various types of dosage forms, and unlike general medicines, it has the advantage of not having side effects that may occur when taking medicines for a long time by using food as a raw material, and has excellent portability.

Another aspect of the present invention provides a feed composition for preventing or improving menopausal diseases comprising a mixed extract or fractions of Santianmok and Yangjegeun.

The feed composition may include a feed additive. The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act.

The term "feed" in the present invention may refer to any natural or artificial diet, one meal, etc., or a component of the one meal meal, suitable for or suitable for eating, ingestion and digestion by an animal.

The type of feed is not particularly limited, and feeds commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products; Animal feed such as proteins, inorganic materials, oils, mineral oils, oils, single cell proteins, zooplankton, or food may be mentioned. These may be used alone or in combination of two or more.

The composition containing the mixed extract of Santenmok and Yangjegeun provided in the present invention can effectively improve various symptoms such as reduced levels of ALP, TC, LDL and osteocalcin in the blood, symptoms of depression, and increased fat cell size in menopausal diseases. Therefore, it will be widely used in the development of therapeutic agents for menopausal diseases.

Figure 1a is a graph showing the results of comparison of average body weight change according to the lapse of breeding period for each experimental group provided by the present invention.
Figure 1b is a graph showing the results of comparing the weight of adipose tissue for the experimental animals for each experimental group provided by the present invention.
Figure 2a is a graph showing the results of comparing the levels of ALP (alkaline phosphatase) contained in serum derived from laboratory animals for each experimental group provided by the present invention.
Figure 2b is a graph showing the results of comparing the levels of total cholesterol (TC) contained in serum derived from laboratory animals for each experimental group provided by the present invention.
Figure 2c is a graph showing the results of comparing the level of low density lipoprotein (LDL) contained in serum derived from laboratory animals for each experimental group provided by the present invention.
Figure 3 is a graph showing the results of comparison of the concentration of osteocalcin contained in serum derived from laboratory animals for each experimental group provided by the present invention.
Figure 4a is a graph showing the results of comparing the moving distance and moving time of the experimental animals for each experimental group provided by the present invention in the center (cemter) and edge (periphe) areas in the open field box.
Figure 4b is a schematic diagram illustrating the trajectory of the movement of the experimental animal for each experimental group provided by the present invention in the open field box.
5a is a photomicrograph showing adipocytes included in adipose tissue of experimental animals for each experimental group provided by the present invention.
5B is a graph showing the result of comparing the sizes (diameters) of the adipocytes.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

Example 1: Production of ovariectomy animal model

8-week-old female mice (C57BL/6J) were purchased from Orient Bio Co., Ltd. and acclimatized in a quarantine room for one week. Subsequently, the mouse was anesthetized to sterilize the surgical part, and then the surgical part was incised and the ovary was pulled and exposed. Then, after fixing the ovary by fixing the lower end of the ovary, hemostasis was performed. An ovariectomy animal model (OVX) was produced by incising the hemostasis in the ovary, checking for bleeding, and then suturing the incision.

Example 2: Preparation of samples

First, Santenmok was immersed in distilled water and heated for 4 hours, and then filtered to obtain a liquid component. ) was obtained.

Next, a mixture of 6:2:1:1 (w/w/w/w) of Santenmok, Duchung, Ussul, and Angelica is immersed in distilled water, heated for 4 hours, and then filtered to obtain a liquid component. and concentration under reduced pressure and lyophilization were sequentially performed on the obtained liquid component to obtain a composite extract of Santenmok (ATT).

Finally, a mixture of Sancheonmok and Yangjegeun mixed at a ratio of 6:4 (w/w) was immersed in distilled water and heated for 4 hours, and then filtered to obtain a liquid component. Concentration and freeze-drying were sequentially performed to obtain a mixed extract (RC) of Sancheonmok-Yangjegeun.

Example 3: Administration of Samples

Each sample prepared in Example 2 was orally administered to the ovariectomy animal model (OVX) prepared in Example 1.

First, each of the extracts prepared in Example 2 was dissolved in distilled water to obtain a liquid sample, which was orally administered to OVX, respectively. At this time, oral administration is carried out under the same breeding environment (temperature 23±3℃, relative humidity 55±15%, ventilation frequency 10-20 times/hr, lighting time 12 hours and illumination 150-300 Lux), once a day in the morning. It was performed by administering 200 μL of liquid sample for 10 weeks using a sonde.

In addition, the normal control group (Normal) used mice without ovariectomization to which only distilled water was orally administered, the negative control group (OVX) used OVX to which only distilled water was orally administered, and the positive control group (OVX + EST) was 17- OVX with intravenous administration of beta-estradiol was used.

Finally, the dosage of the sample was set to the conditions in Table 1 below.

Setting up the experimental group experimental group laboratory animal sample dose Normal
OVX
OVX+EST
OVX+AT100
OVX+AT200
OVX+ATT200
OVX+RC100
OVX+RC200 normal mouse
OVX
OVX
OVX
OVX
OVX
OVX
OVX Distilled water
Distilled water
estradiol
AT
AT
ATT
RC
RC -
-
-
100 mg/kg
200 mg/kg
200 mg/kg
100 mg/kg
200 mg/kg

Example 4: Weight change of body weight and adipose tissue

In Example 3, the change in average body weight for each experimental group was measured by measuring the change in body weight of each experimental animal raised during oral administration for 10 weeks once a week (FIG. 1a).

Figure 1a is a graph showing the results of comparison of average body weight change according to the lapse of breeding period for each experimental group provided by the present invention.

As shown in Figure 1a, it was confirmed that the body weight increased with the passage of the breeding period as a whole.

In addition, it was confirmed that the weight of the negative control group (OVX) significantly increased compared to the weight of the normal control group (Normal), while the weight of the positive control group (OVX+EST) significantly decreased compared to the weight of the negative control group (OVX).

In addition, the body weight of the remaining experimental groups (OVX+AT100, OVX+AT200, OVX+ATT200, OVX+RC100 and OVX+RC200) was reduced compared to that of the negative control group (OVX), but it was confirmed that there was no significant difference.

In addition, after breeding was performed for 10 weeks, the weights of adipose tissue isolated from each of the experimental animals were compared (Fig. 1b).

Figure 1b is a graph showing the results of comparing the weight of adipose tissue for the experimental animals for each experimental group provided by the present invention.

As shown in Figure 1b, the fat weight of the negative control group (OVX) increased significantly compared to the fat weight of the normal control group (Normal), whereas the fat weight of the positive control group (OVX + EST) compared to the fat weight of the negative control group (OVX). It was confirmed that the weight was significantly reduced.

In addition, among the remaining experimental groups (OVX+AT100, OVX+AT200, OVX+ATT200, OVX+RC100 and OVX+RC200), the fat weight of OVX+RC100 and OVX+RC200 decreased significantly compared to that of the negative control group (OVX). However, it was confirmed that the fat weight of the other experimental groups (OVX+AT100, OVX+AT200 and OVX+ATT200) did not show a significant difference from that of the negative control group (OVX).

Example 5: Serology analysis

After breeding for 10 weeks in Example 3, blood was collected from the experimental animals of each experimental group, the collected blood was coagulated, and then centrifuged (1,500 g and 10 minutes) to obtain serum.

Subsequently, the obtained serum was applied to an Automated analyzer (7080, Hitachi Ltd., Tokyo, Japan) to measure the levels of Alkaline phosphatase (ALP), total cholesterol (TC) and low density lipoprotein (LDL) contained in the serum. was measured (Figs. 2a to 2c).

Figure 2a is a graph showing the results of comparing the levels of ALP (alkaline phosphatase) contained in serum derived from laboratory animals for each experimental group provided by the present invention.

As shown in Figure 2a, compared to the measured value of the normal control group (Normal), the measured value of the negative control group (OVX) increased significantly, whereas the measured value of the negative control group (OVX) compared to the measured value of the positive control group (OVX + EST) It was confirmed that the measured value was significantly reduced.

In addition, the measured values of the remaining experimental groups (OVX+AT100, OVX+AT200, OVX+ATT200, OVX+RC100 and OVX+RC200) were also reduced to a level showing a statistically significant difference compared to the measured values of the negative control group (OVX). .

In particular, it was found that OVX+RC100 and OVX+RC200 decreased to a significantly lower level than other experimental groups.

Figure 2b is a graph showing the results of comparing the levels of total cholesterol (TC) contained in serum derived from laboratory animals for each experimental group provided by the present invention.

As shown in FIG. 2B, the measured value of the negative control group (OVX) increased significantly compared to the measured value of the normal control group (Normal), whereas the measured value of the positive control group (OVX + EST) compared to the measured value of the negative control group (OVX). It was confirmed that the measured value was significantly reduced.

In addition, the measured values of the remaining experimental groups (OVX+AT100, OVX+AT200, OVX+ATT200, OVX+RC100 and OVX+RC200) were also reduced to a level showing a statistically significant difference compared to the measured values of the negative control group (OVX). .

In particular, it was found that OVX+RC100 and OVX+RC200 decreased to a significantly lower level than other experimental groups.

Figure 2c is a graph showing the results of comparing the level of low density lipoprotein (LDL) contained in serum derived from laboratory animals for each experimental group provided by the present invention.

As shown in Figure 2c, compared to the measured value of the normal control group (Normal), the measured value of the negative control group (OVX) increased significantly, whereas the measured value of the negative control group (OVX) compared to the measured value of the positive control group (OVX + EST) It was confirmed that the measured value was significantly reduced.

In addition, the measured values of the remaining experimental groups (OVX+AT100, OVX+AT200, OVX+ATT200, OVX+RC100 and OVX+RC200) were also reduced to a level showing a statistically significant difference compared to the measured values of the negative control group (OVX). .

In particular, it was found that OVX+RC100 and OVX+RC200 decreased to a significantly lower level than other experimental groups.

Example 6: Blood osteocalcin concentration analysis

Since osteocalcin increases during menopause and is used as a marker for early diagnosis of postmenopausal osteoporosis, the concentration of osteocalcin in blood contained in serum derived from laboratory animals for each experimental group obtained in Example 7 was compared and analyzed.

Roughly, each serum sample was put in a 96-well plate coated with anti-mouse osteocalcin monoclonal antibody included in the enzyme immunoassay set (Takara Bio Inc.), reacted at room temperature for 1 hour, and then secondary peroxidase-labeled anti- After adding Gla-Osteocalcin monoclonal antibody, it was reacted at room temperature for 1 hour. Finally, after adding Substrate Solution (TMBZ) to the plate and reacting for 30 minutes, the reaction was terminated by adding TMBZ stop solution. Optical density was measured for absorbance at 450 nm using a microplate reader (SoftMax PRO, version 3.1 software, CA, USA) (FIG. 3).

Figure 3 is a graph showing the results of comparison of the concentration of osteocalcin contained in serum derived from laboratory animals for each experimental group provided by the present invention.

As shown in FIG. 3, the concentration of osteocalcin in the negative control group (OVX) was significantly increased compared to the concentration of osteocalcin in the normal control group (Normal), whereas the concentration of osteocalcin in the positive control group (OVX+) was significantly increased compared to the concentration of osteocalcin in the negative control group (OVX). EST) was found to significantly decrease the concentration of osteocalcin.

In addition, the concentration of osteocalcin in the remaining experimental groups (OVX + AT100, OVX + AT200, OVX + ATT200, OVX + RC100 and OVX + RC200) was also reduced to a level showing a statistically significant difference compared to the concentration of osteocalcin in the negative control group (OVX). Confirmed.

In particular, it was found that OVX+RC100 and OVX+RC200 decreased to a significantly lower level than other experimental groups.

Example 7: Depression Assay

In order to compare the level of abnormal activity of mice according to the degree of depression of experimental animals for each experimental group provided in the present invention, according to a known method (open field test), using video tracking equipment in an open field box The behavior of the mice was observed for 30 minutes.

Roughly, the movement of each experimental animal is monitored using infrared detectors installed at 2.54 cm intervals, and ambulatory activity, stereotypic activity, and rearing are recorded for frequency and time. analyzed.

As is known, the behavioral patterns that appear when exposing an experimental animal to a new space are innate behaviors and are distinguished from general learned behaviors. In this embodiment, the total activity level for 5 minutes is analyzed. It was observed separately for each factor (parameters), and also exposed to the same activity measurement box twice at 24-hour intervals to examine and compare the memory and learning level for the environment through behavioral analysis (FIG. 4a and 4b).

Figure 4a is a graph showing the results of comparing the moving distance and moving time of the experimental animals for each experimental group provided in the present invention in the center (cemter) and edge (periphe) areas in the open field box, Figure 4b is a graph in the present invention It is a schematic diagram showing the movement trajectory of the experimental animals for each experimental group provided in the open field box.

As shown in FIGS. 4a and 4b, the normal control group (Normal) shows active movement targeting the entire area of the open field box, whereas the negative control group (OVX) shows depression staying in the periphery area rather than the center area (cemter). It was confirmed that a depression-like behavior was exhibited.

However, the positive control group (OVX+EST) showed active movement in the entire area of the open field box at a similar level to the normal control group (Normal), and the remaining experimental groups (OVX+AT100, OVX+AT200, OVX+ATT200, OVX +RC100 and OVX+RC200) also showed more active movements than the negative control group (OVX). I was able to confirm.

Example 8: Histological analysis of the uterus

After performing breeding for 10 weeks in Example 3, uterine tissue was extracted from the experimental animals of each experimental group, and then placed in 10% neutral formalin and fixed for 24 hours. Then, the fixed tissue was processed in a conventional manner to obtain tissue slices having a thickness of 4 μm, and the obtained tissue slices were stained with hematoxylin and eosin (H&E). Stained tissue sections were photographed under a microscope, and digital images were obtained using Leica Application Suite LAS microscope software (Leica Microsystems, Buffalo Grove, IL, USA). The size (diameter) of adipocytes was measured using Image J (NIH, Bethesda, MD, USA) (FIGS. 5a and 5b).

Figure 5a is a photomicrograph showing fat cells included in adipose tissue of experimental animals for each experimental group provided by the present invention, and Figure 5b is a graph showing the result of comparing the sizes (diameters) of the fat cells.

As shown in FIGS. 5A and 5B, the measured values of the negative control group (OVX) were significantly increased compared to the measured values of the normal control group (Normal), whereas the measured values of the negative control group (OVX) were compared to the measured values of the positive control group (OVX+ EST) was confirmed to be significantly reduced.

In addition, among the remaining experimental groups (OVX+AT100, OVX+AT200, OVX+ATT200, OVX+RC100 and OVX+RC200), only the measured values of OVX+RC100 and OVX+RC200 were statistically more significant than the negative control group (OVX). It was confirmed that it was reduced to a level showing a difference.

From the above description, those skilled in the art to which the present invention pertains will be able to understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, the embodiments described above should be understood as illustrative in all respects and not limiting. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the claims to be described later and equivalent concepts rather than the detailed description above are included in the scope of the present invention.

Claims (10)

A pharmaceutical composition for the prevention or treatment of female menopausal diseases comprising a mixed extract of Santianmok and Yangjegeun,
The female menopausal disease is selected from the group consisting of weight gain, facial flushing, headache, gastrointestinal disorders, depression, vaginal dryness, and combinations thereof, the pharmaceutical composition.
According to claim 1,
The pharmaceutical composition, wherein the mixing ratio of Sancheonmok and Yangjegeun is 9: 1 to 1: 9 (w / w).
According to claim 1,
The mixed extract is obtained by extracting a mixture of Santianmok and Yangjegeun or obtained by mixing Santianmok extract and Yangjegeun extract.
delete delete According to claim 1,
The extraction is performed by a hot water extraction method, an ultrasonic extraction method, a room temperature extraction method, a cold extraction method, a reflux cooling extraction method, or a steam extraction method, a pharmaceutical composition.
delete A method for preventing or treating menopausal diseases, comprising administering the pharmaceutical composition according to any one of claims 1 to 3 and 6 to a non-human subject in need of prevention or treatment of women's menopausal diseases.
A health functional food composition for preventing or improving women's menopausal diseases, comprising a water mixture extract of Sancheonmok and Yangjegeun,
The female menopausal disease is selected from the group consisting of weight gain, facial flushing, headache, gastrointestinal disorders, depression, vaginal dryness, and combinations thereof, health functional food composition.
A feed composition for preventing or improving menopausal diseases in female mammals containing a water mixture extract of Santianmok and Yangjegeun,
The menopausal disease of the female mammal is selected from the group consisting of weight gain, facial flushing, headache, gastrointestinal disorder, depression, vaginal dryness, and combinations thereof, feed composition.

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