KR20230068357A - Compositions for preventing or treating menopausal syndrome of women comprising cudrania tricuspidata extract, angelica sinensis extract and lycii radicis cortex extract as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of menopausal symptoms containing a mixed extract of Cudrania tricuspidata, Angelica gigas, and Lycium chinense as an active component. Specifically, the mixed extract mixing extracts of Cudrania tricuspidata, Angelica gigas, and Lycium chinense has estrogenic activity, and confirms that the extract acts as an estrogen receptor agonist without side effects, thereby being usefully used in female hormone replacement therapy to prevent and treat menopausal symptoms caused by estrogen deficiency in menopausal women.

Description

Composition for preventing or treating menopausal symptoms containing kujippong extract, angelica extract, and gigolpi extract as active ingredients

The present invention relates to a pharmaceutical composition for preventing or treating menopausal symptoms containing an extract of Cudrania tricuspidata , an extract of Angelica sinensis , and an extract of Lycii Radicis Cortex as active ingredients, and a health functional food for preventing or improving menopausal symptoms It is about.

Women experience menopausal symptoms gradually, 10 years before and after menopause, usually between the ages of 45 and 55, and suffer from physical and mental difficulties. According to the statistics of the Health Insurance Review and Assessment Service, about 430,000 patients visited the hospital with menopausal symptoms as of 2013. is gradually decreasing, so it is necessary to manage women's menopausal symptoms.

Menopause is a type of endocrine syndrome, and refers to a transitional period in which physiological and sexual functions are reduced or lost due to a decrease in female hormones, that is, estrogen, caused by overall and gradual aging of ovarian function. These menopausal symptoms include symptoms due to vascular changes such as hot flashes, tachycardia, sweating or headaches, symptoms due to musculoskeletal changes such as muscle pain, joint pain and back pain, and symptoms due to genitourinary changes such as frequent urination or urinary incontinence. There are symptoms caused by changes in the brain nervous system such as memory loss, depression, loss of concentration, and dizziness. In addition, symptoms such as vision loss and skin and hair changes occur, and women such as osteoporosis or cardiovascular disease due to hormonal changes diseases that can be fatal to your health.

Therefore, in order to improve the physical and mental health and quality of life of middle-aged women, the development of therapeutic agents capable of improving menopausal symptoms has been required, and drugs such as hormone replacement therapy and non-steroidal agents have been developed to improve these menopausal symptoms. has been However, most of these drugs are known to have side effects such as headaches and weight gain. In particular, even in the case of estrogen replacement therapy, since hormones are artificially administered into the body, they are rejected along with uterine bleeding, stroke, and heart disease. It is known that it may increase the risk of developing seizures, breast and uterine cancer.

Because of these problems, interest in replacing estrogen therapy with natural methods ingested in the form of food or additives is increasing, and the development of new menopausal symptom treatments that have excellent effects in relieving menopausal symptoms without side effects is required. There is a situation.

On the other hand, estrogen has various biological functions including cell number increase and differentiation. Biological actions of estrogen exert their activity through two estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ) belonging to the nuclear receptor superfamily. The mechanism of action of the estrogen receptor is the combination of estrogen and intracellular estrogen receptors to form a dimer of the receptors, which are located in the promoters of the target gene. Specific estrogen response elements (EREs) ) to regulate the expression of target genes. Since the two estrogen receptors have different tissue distribution patterns and ligand binding capacity, these differences enable different selective roles of agonists and antagonists of the estrogen receptor in different tissues. In addition, substances acting on estrogen receptors are being used as targets for hormone replacement therapy for postmenopausal women and chemotherapy for genital cancer. Therefore, if a natural product that activates the estrogen receptor is discovered for a natural product with few side effects on the human body, it can be usefully used for the treatment and prevention of female menopausal symptoms, so related research is steadily progressing.

Therefore, as a result of efforts to develop a combination drug for treating female menopausal symptoms derived from natural products that is safe for the human body, the mixed extract of kkujippong extract, Angelica gigas extract, and gigolpi extract has estrogenic activity and acts as an estrogen receptor agonist without side effects. By confirming that, the present invention was completed by revealing that the mixed extract can be used as a female hormone replacement therapy for preventing and treating menopausal symptoms caused by estrogen deficiency in menopausal women.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating female menopausal symptoms, containing Cudrania tricuspidata extract, Angelica sinensis extract, and Lycii Radicis Cortex extract as active ingredients.

Another object of the present invention is to provide a health functional food for preventing or improving women's menopausal symptoms, containing kkujippong extract, angelica extract, and gigolpi extract as active ingredients.

Another object of the present invention is to provide a food composition for preventing or improving female menopausal symptoms, containing kkujippong extract, Angelica gigas extract, and gigolpi extract as active ingredients.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating female menopausal symptoms, containing Cudrania tricuspidata extract, Angelica sinensis extract, and Lycii Radicis Cortex extract as active ingredients. do.

The present invention provides a health functional food for preventing or improving female menopausal symptoms, containing kkujippong extract, angelica extract, and gigolpi extract as active ingredients.

The present invention provides a food composition for preventing or alleviating female menopausal symptoms, containing kkujippong extract, angelica extract, and gigolpi extract as active ingredients.

The mixed extract of the kkujippong extract, angelica extract, and gigolpi extract of the present invention has estrogenic activity and acts as an estrogen receptor agonist without side effects, so it prevents and treats menopausal symptoms caused by estrogen deficiency in menopausal women It can be used for female hormone replacement therapy for

1 is an estrogen receptor-positive breast cancer cell line MCF-7 treated with kkujippong extract, angelica extract, gigolpi extract, or a mixed extract of kkujippong, angelica and gigolpi prepared by mixing in a mixing ratio showing excellent effects according to an embodiment of the present invention It is a diagram confirming the synergistic effect of the mixed extract of kkujippong, Angelica gigas and gigolpi of the present invention by comparing ER-mediated transcriptional activity.
Figure 2 is an estrogen receptor-positive breast cancer cell line MCF-7 treated with kkujippong extract, angelica extract, gigolpi extract, or mixed extract of kkujippong, angelica and gigolpi prepared by mixing in a mixing ratio showing excellent effects according to an embodiment of the present invention It is a diagram confirming the synergistic effect of the estrogen-like activity of the mixed extract of angelica gigas and gigolpi of the present invention by comparing the cell proliferation rate after treatment.
Figure 3 compares ER-mediated transcriptional activity after treating endometrial cancer adenocarcinoma cell line Ishikawa with mixed extracts of kkujippong, angelica and gigolpi prepared by mixing at a mixing ratio showing excellent effects according to an embodiment of the present invention by concentration. It is a diagram confirming the presence or absence of uterine growth side effects of the mixed extract of kkujippong, angelica and gigolpi of the present invention.

Hereinafter, the present invention will be described in more detail.

The present invention provides a pharmaceutical composition for preventing or treating female menopausal symptoms, containing Cudrania tricuspidata extract, Angelica sinensis extract, and Lycii Radicis Cortex extract as active ingredients.

Each of the active ingredients of the present invention, kkujippong extract, Angelica gigas extract, and gigolpi extract, is preferably prepared by a method comprising the following steps, but is not limited thereto:

1) extracting by adding an extraction solvent to each of kkujippong, angelica and jigolpi; and

2) Filtering each of the kkujippong extract, Angelica gigas extract, and gigolpi extract of step 1).

In the present invention, the kkujippong, angelica and gigolpi of step 1) can be used without limitation cultivated or commercially available. In addition, flowers, branches, stems, leaves, fruits, aerial parts, rhizomes, roots, or combinations thereof of the kkujippong and Angelica quai may be used, but are not limited thereto, and the phalanx is the root of Lycium chinense Miller or other plants of the same genus It is preferable that the peel is dried.

In the present invention, it is preferable to use water, alcohol or a mixture thereof as the extraction solvent in step 1). It is preferable to use a C ₁ to C ₂ lower alcohol as the alcohol, and it is preferable to use ethanol or methanol as the lower alcohol. As an extraction method, it is preferable to use ultrasonic extraction, shaking extraction, Soxhelt extraction or reflux extraction, but is not limited thereto. It is preferable to extract by adding 1 to 15 times the amount of washed and well-dried kkujippong, angelica and gigolpi with the extraction solvent, and more preferably to extract by adding 2 to 10 times. The extraction temperature is preferably 20 to 110 ° C, but is not limited thereto. In addition, the extraction time is preferably 10 minutes to 72 hours, more preferably 20 minutes to 48 hours, but is not limited thereto. In addition, the number of times of extraction is preferably 1 to 5 times, but is not limited thereto.

In the present invention, the kkujippong extract, angelica extract and gigolpi extract can be mixed after preparing each extract, but it is also possible to prepare in the form of obtaining an extract thereof after mixing kkujippong, angelica gigolica and gigolpi.

In the present invention, the following steps may be additionally included after step 2):

3) concentrating each of the filtrate of step 2) under reduced pressure; and

4) drying each of the concentrates of step 3).

In the present invention, the vacuum concentration in step 3) is preferably performed using a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, drying is preferably performed under reduced pressure, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.

In the present invention, after mixing kkujippong, angelica and gigolpi, an extract thereof may be obtained, and concentrated under reduced pressure and dried.

In the present invention, the composition may contain kkujippong extract: angelica extract: jigolpi extract in a weight ratio of 1: 1 to 1000: 1 to 500, and more specifically, a weight ratio of 1: 300 to 800: 50 to 400. And, more specifically, it may be contained in a weight ratio of 1:400 to 750:100 to 300.

In the present invention, the female menopausal symptoms collectively refer to symptoms and diseases that occur in women before and after menopause as the secretion of estrogen decreases due to ovarian aging, and hot flushes, sweating, dry skin, vaginal dryness, and vaginal atrophy , lower urethral atrophy, vaginitis, cystitis, dysuria, urgency to urinate, difficulty concentrating, short-term memory impairment, anxiety, nervousness, memory loss, muscle pain, arthralgia, or osteoporosis, but is not limited thereto.

In the present invention, the composition has estrogenic activity and exhibits estrogen receptor (ER) agonist activity, thereby having a therapeutic effect on female menopausal symptoms.

In a specific embodiment of the present invention, the present inventors prepared kkujippong extract, angelica extract and gigolpi extract, and mixed the extracts in a specific mixing ratio to prepare kkujippong, angelica and gigolpi extracts.

In addition, the present inventors measured the degree of ER-mediated transcriptional activity by treating the estrogen receptor-positive breast cancer cell line MCF-7 with mixed extracts of kkujippong, angelica and gigolpi, and as a result, compared to the case of treating kkujippong, angelica and gigolpi extracts respectively, kkujippong Extract: Angelica extract: When the extract of Angelica gigolica was mixed at a mixing ratio of 1: 400 to 750: 100 to 300, it was confirmed that the activity increased more than the sum of the ER-mediated transcriptional activities of each extract, showing a synergistic effect (FIG. 1 reference).

In addition, the present inventors measured the cell viability by treating the mixed extract with the estrogen receptor-positive breast cancer cell line MCF-7, and compared the sum of the cell viability treated with each of the extracts of kkujippong, Angelica and gigolpi. Treatment of the mixed extract It was confirmed that one cell viability increased, indicating a synergistic effect (see FIG. 2).

In addition, the present inventors administered the mixed extract to the endometrial cancer adenocarcinoma cell line Ishikawa at different concentrations and measured the degree of ER-mediated transcriptional activity. Since it was significantly smaller than the result, it was confirmed that there was almost no uterine hyperplasia symptom, one of the side effects of the ER agonist (see FIG. 3).

Therefore, since the mixed extract of kkujippong, Angelica gigas and gigolpi of the present invention has estrogenic activity and exhibits activity as an ER agonist without side effects, the extract of the present invention, kkujippong extract, angelica gigas extract, and gigolpi extract can be used to treat menopausal symptoms caused by estrogen deficiency in menopausal women. It can be usefully used as an active ingredient of a pharmaceutical composition for preventing or treating.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The pharmaceutical composition according to the present invention can be administered orally or parenterally, and in the case of parenteral administration, external skin or intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method is selected. It is preferred, but is not limited thereto.

The pharmaceutical composition according to the present invention is formulated in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods, respectively. can be used Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain at least one or more excipients such as starch, calcium carbonate, sucrose in mixed herbal medicines. ) or by mixing lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, solutions for oral use, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried 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. In addition, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary.

A preferred dosage of the pharmaceutical composition according to the present invention varies depending on the degree of absorption of the active ingredient in the body, the age, sex and severity of symptoms of the patient, but can be appropriately selected by those skilled in the art. However, for the desired effect, in the case of oral administration, it is generally recommended to administer the composition of the present invention at 0.0001 to 1000 mg/kg per day, preferably 0.001 to 500 mg/kg per 1 kg of body weight per day to adults. good night. Administration may be administered once a day, or may be administered in several divided doses. The dosage is not intended to limit the scope of the present invention in any way.

The present invention provides a health functional food for preventing or improving female menopausal symptoms, containing kkujippong extract, angelica extract, and gigolpi extract as active ingredients.

The kkujippong extract, angelica extract, and gigolpi extract, their mixing ratio, and female menopausal symptoms are the same as the description of the pharmaceutical composition for preventing or treating female menopausal symptoms. Only the specific composition of nutraceuticals should be described.

On the other hand, since the mixed extract of kkujippong, Angelica gigas and gigolpi of the present invention has estrogenic activity and exhibits activity as an ER agonist without side effects, the mixed extract of the present invention mixed with kkujippong extract, angelica extract and gigolpi extract is estrogen in menopausal women It can be usefully used as an active ingredient in health functional foods to prevent or improve menopausal symptoms caused by deficiency.

The three extracts of the present invention can be provided as a food composition by mixing with a carrier acceptable for food science.

When the three extracts of the present invention are used as food or beverage additives, the three extracts may be added as they are or used together with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the three extracts may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment). In the case of long-term intake for the purpose of health and hygiene or health control, the three extracts can be taken for a long period of time because there is no problem in terms of safety. There is no particular limitation on the type of food.

Examples of foods to which the above substances can be added include meat, sausages, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, and drinks. , alcoholic beverages and vitamin complexes. When formulated into a beverage, liquid components added in addition to the above three extracts are not limited now, but may contain various flavors or natural carbohydrates as additional components, as in conventional beverages. Examples of the aforementioned natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.) and polysaccharides (eg, common sugars such as dextrins, cyclodextrins, etc.), and xylitol. , sorbitol, erythritol, etc. are sugar alcohols. The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention. As flavoring agents other than those mentioned above, natural flavoring agents [eg, taumarin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)] and synthetic flavoring agents (eg, saccharin, aspartame, etc.) can be used. .

In addition, the food composition of the present invention contains various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, organic acids, protective It may contain a colloidal thickener, a pH adjusting agent, a stabilizer, a preservative, glycerin, alcohol, a carbonating agent used in carbonated beverages, and the like. In addition, the food composition of the present invention may contain fruit flesh for preparing fruit and vegetable beverages. These components may be used alone or in combination, and the proportion of these additives is generally selected from the range of 0.001 to 50 parts by weight per total weight of the composition.

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

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

<Example 1> Kkujippong ( Cudrania tricuspidata ) preparation of the extract

The obtained kkujippong ( Cudrania tricuspidata ) The fruit was completely frozen at minus 20 ° C., dried using a freeze dryer and then pulverized with a grinder. After adding 10 ml of 70% methanol to 2 g of the lyophilized powder of the kkujippong fruit, ultrasonic extraction was performed for 15 minutes. The ultrasonically extracted kkujippong extract was centrifuged at 3,000 rpm for 10 minutes, and then the supernatant was separated and filtered with a 0.45 μm PTFE-syringe filter.

<Example 2> Angelica ( Angelica sinensis ) preparation of the extract

The obtained Angelica sinensis root was completely frozen at minus 20 ° C, dried using a lyophilizer and then pulverized with a grinder. 10 ml of 70% methanol was added to 2 g of the lyophilized powder of Angelica root, followed by ultrasonic extraction for 15 minutes. The ultrasonically extracted kkujippong extract was centrifuged at 3,000 rpm for 10 minutes, and then the supernatant was separated and filtered with a 0.45 μm PTFE-syringe filter.

<Example 3> Phalangeal skin ( Lycii Radicis Cortex ) preparation of the extract

The obtained phagocytic ( Lycii Radicis Cortex ) was completely frozen at minus 20 ° C, dried using a lyophilizer and then pulverized with a grinder. After adding 10 ml of 70% methanol to 2 g of the lyophilized powder of the phalanx, ultrasonic extraction was performed for 15 minutes. The ultrasonically extracted kkujippong extract was centrifuged at 3,000 rpm for 10 minutes, and then the supernatant was separated and filtered with a 0.45 μm PTFE-syringe filter.

<Example 4> Preparation of mixed extracts of kkujippong, Angelica gigas and gigolpi

The kkujippong extract prepared in <Example 1>, the Angelica gigas extract prepared in <Example 2>, and the Angelica gigolica extract prepared in <Example 3> were mixed according to the composition shown in the following [Table 1] to mix kkujippong, angelica gigolica and gigolpi. An extract was prepared.

Example Composition of extract mixture (μg/ml) Extract mixing ratio kkujippong angelica phalanx Cudrania: Angelica: Jigolpi 4 3 2000 500 0.0015:1:0.25

<Experimental Example 1> Confirmation of estrogen receptor (ER) agonist activity of mixed extracts of kkujippong, Angelica gigas and gigolpi

The biological action of estrogen is mediated by the binding of estrogen to the estrogen receptor (ER) as a ligand, and since the ER acts as a transcription factor and binds to ERE (estrogen response element), an ERE reporter, that is, ER-reactive luciferin It is known that the effect of a therapeutic agent can be monitored by confirming ER-mediated transcriptional activity using the enzyme. Therefore, in order to investigate whether the mixed extract of kkujippong, Angelica gigas and gigolpi could improve menopausal symptoms caused by estrogen deficiency in menopausal women by acting as an ER agonist, a specific mixing ratio was applied to cells transfected with a plasmid containing ERE-luciferase. The degree of ER-mediated transcriptional activity was measured after treating the mixed extracts of kkujippong, Angelica gigas and Jigolpi prepared by mixing with .

Specifically, the estrogen receptor-positive breast cancer cell line MCF-7 purchased from the American Type Culture Collection (ATCC) was cultured in DMEM (Dulbecco's modified Eagle's medium) medium containing non-essential amino acids, antibiotics/antibiotics. /antimycotics), sodium pyruvate and 10% fetal bovine serum were added as a basal medium to culture the cells. They were subcultured once a week on average. 48 to 60 hours (2 to 3 days) before cell seeding, serum was treated with dextran-charcoal three times, and it contained stripped serum containing phenol-red. The cells were cultured using a medium prepared with DMEM (Dulbecco's modified Eagle's medium) without the This medium was continuously used until the end of the luciferase assay. The reason for using the medium is to exclude the estrogen properties of phenol-red itself and the result of estrogen receptor interaction by hormones and growth factors contained in serum in large amounts.

Then, the cultured breast cancer cell line MCF-7 was inoculated (10 ⁵ cells/well) in a 24-well plate one day before transfection, and then ERE-luciferase plasmid DNA (0.25 μg/well) was added to Lipofectamine 2000 The breast cancer cell line was transfected using a transfection reagent (Invitrogen).

As a comparative group, the kkujippong extract prepared in <Example 1>, the Angelica gigas extract prepared in <Example 2>, and the Jigolpi extract prepared in <Example 3> were used, and the experimental group prepared in <Example 4> Mixed extracts of kkujippong, angelica and gigolpi were used, and each extract was tested at a concentration of 200 μg/ml. On the other hand, the cells were treated with mixed extracts of kkujippong, Angelica gigas, and gigolpi, and cultured for an additional 24 hours, followed by luciferase assay. After removing the medium, the cells were cultured in cold PBS for 2 After washing twice, the cell-soluble material (lysate) was extracted with 1X passive lysis buffer (Promega), and 50 μl of cell-soluble material was mixed with 50 μl of luciferase assay reagent (including luciferin, which is a substrate of luciferase, and a buffer suitable for enzymatic reaction). After mixing the solution, Promega), the generated luminescence intensity was measured using SpectraMax M5 ^e (Molecular Device). In addition, the % increase value (% Increase) of the ER-mediated transcriptional activity of the Kkujippong, Angelica, and Jigolpi mixed extract treatment group compared to the sum of the ER-mediated transcriptional activity of each of the control groups was calculated using the following equation: %Increase = [(The degree of ER-mediated transcriptional activity of the mixed extract treatment group of Kujippong, Angelica gigas, and Jigolpi - the sum of the levels of ER-mediated transcriptional activity of each control group) / the sum of the levels of ER-mediated transcriptional activity of each control group] × 100.

As a result, as shown in Figure 1, the % increase value of the mixed extract treatment group was 21.2%, and the kkujippong extract treatment group, angelica extract treatment group, and gigolpi extract treatment group treated at the same concentration as the concentration contained in the mixed extract, respectively. It was confirmed that the degree of ER-mediated transcriptional activity of the mixed extract treatment group was significantly increased compared to the sum of the levels of ER-mediated transcriptional activity of (FIG. 1).

Through the above results, it was confirmed that the mixed extract of the present invention exhibited a synergistic effect as an ER agonist, including all of the kkujippong extract, angelica extract, and gigolpi extract.

<Experimental Example 2> Confirmation of estrogen-like activity of mixed extracts of kkujippong, Angelica gigas and gigolpi

Estrogenic activity is known to proliferate cells in which the estrogen receptor (ER) is endogenously expressed. Therefore, in order to find out whether the mixed extract of kkujippong, Angelica gigas and gigolpi can have an action similar to estrogen, the breast cancer cell line MCF-7 was treated with the mixed extract of kkujippong, angelica gigolica and gigolpi, and then the degree of cell proliferation was measured.

Specifically, the breast cancer cell line MCF-7 purchased from the American Type Culture Collection (ATCC) was seeded in a 24-well plate and cultured for 24 hours, followed by dimethylsulfoxide (DMSO) in the control group. , In the comparative group, the kkujippong extract prepared in <Example 1>, the Angelica gigas extract prepared in <Example 2>, and the phalanx extract prepared in <Example 3> were used, and in the experimental group, the extract prepared in <Example 4> was used. A mixed extract of kkujippong, angelica and gigolpi was used, and each extract was tested at a concentration of 200 μg/ml. Then, on the 1st, 3rd, and 5th days after inoculation, cells were treated with each extract, and on the 7th day, MTT assay was performed to quantify the number of surviving cells. 0.5 mg/ml of silver MTT, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, and yellow tetrazole were dispensed into each well by 10 μl, then the medium was removed after 2 hours. , After adding 100 μl of DMSO, dissolving blue formazan crystal, it was measured at 595 nm using SpectraMax M5 ^e (Molecular Device). In addition, the % increase in MCF-7 cell proliferation (%Increase) of the mixed extract of kkujippong, Angelica gigas and gigolpi compared to the sum of the cell viability of each control group was calculated using the following formula: %Increase=[ (The degree of MCF-7 cell proliferation in the mixed extract treatment group of Kujippong, Angelica gigas, and Jigolpi - the sum of the degree of MCF-7 cell proliferation in each control group) / the sum of the degree of MCF-7 cell proliferation in each control group] × 100.

As a result, as shown in Figure 2, the % increase value of the mixed extract treatment group was 31.5%, and the kkujippong extract treatment group, angelica extract treatment group, and gigolpi extract treatment group treated at the same concentration as the concentration contained in the mixed extract, respectively. It was confirmed that the degree of cell proliferation of the mixed extract treatment group was significantly increased compared to the sum of the degree of cell proliferation of (Fig. 2).

Through the above results, it was confirmed that the mixed extract of the present invention exhibits a synergistic effect on estrogen-like activity, including all of the kkujippong extract, angelica extract, and gigolpi extract.

<Experimental Example 3> Confirmation of the safety of mixed extracts of kkujippong, angelica and gigolpi

Estrogen activity is known to be directly or indirectly related to mechanisms such as endometrial cell proliferation, and thus uterine hyperplasia is recognized as one of the side effects of estrogen-type compounds. Therefore, in order to find out whether the mixed extract of kkujippong, angelica and gigolpi is safe for human application, the degree of uterine proliferation was measured after administering the mixed extract of kkujippong, angelica and gigolpi to the endometrial cancer adenocarcinoma cell line Ishikawa.

Specifically, breast cancer cell line MCF-7 and endometrial adenocarcinoma cell line Ishikawa purchased from the American Type Culture Collection (ATCC) were seeded in a 24-well plate and cultured for 24 hours, followed by estrogen response element ERE (ERE). ) The plasmid gene was transfected. Then, 125, 250, 500, and 1000 μg of the mixed extract samples of kkujippong, Angelica gigas, and gigolpi of <Example 4> were added, respectively, and after further culturing for 24 hours, luciferase activity evaluation (luciferase assay) was performed. For the evaluation of the luciferase activity, the medium was removed, the cells were washed twice with cold PBS, and the cell-soluble material (lysate) was extracted with 1X passive lysis buffer (Promega), and 50 μl luciferase assay was performed on 50 μl of cell-soluble material After mixing the reagent (a solution containing luciferin, which is a substrate for luciferase, and a buffer suitable for enzyme reaction, Promega), the generated luminescence intensity was measured using SpectraMax M5 ^e (Molecular Device).

As a result, as shown in FIG. 3, although the degree of transcriptional activity of endometrial adenocarcinoma cells Ishikawa increased by concentration, it was confirmed that the degree of increase was about 11 times lower than that of the breast cancer cell line MCF-7. (Fig. 3).

Through the above results, it was confirmed that the mixed extract of kkujippong, angelica gigolica and gigolpi of the present invention was safe as an ER agonist without side effects of uterine growth.

Therefore, through the results of <Experimental Example 1> to <Experimental Example 3>, it was confirmed that the mixed extract of kkujippong, angelica and gigolpi of the present invention has estrogenic activity and acts as an ER agonist without side effects. The mixed extract of phalanges can be used for female hormone replacement therapy to prevent and treat menopausal symptoms caused by estrogen deficiency in menopausal women.

Hereinafter, production examples of each formulation according to the present invention are exemplified. The following preparation examples are intended to aid understanding of the practice of the present invention, but do not mean that the preparation method of the formulation according to the present invention is limited to the following preparation examples.

<Preparation Example 1> Preparation of pharmaceutical formulation

<1-1> Preparation of powder

2 g of mixed extract of the present invention

1 g lactose

A powder was prepared by mixing the above components and filling in airtight bags.

<1-2> Manufacture of tablets

100 mg of mixed extract of the present invention

Microcrystalline Cellulose 100 mg

Lactose 100 mg

Sodium starch glycolate 18 mg

Magnesium stearate 2 mg

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

<1-3> Preparation of capsules

100 mg of mixed extract of the present invention

Corn Starch 100 mg

100 mg milk sugar

Magnesium stearate 2 mg

After mixing the above ingredients, capsules were prepared by filling gelatin capsules according to a conventional capsule preparation method.

<1-4> Preparation of ring

1 g of mixed extract of the present invention

lactose 1.5 g

1 g of glycerin

0.5 g xylitol

After mixing the above components, it was prepared to be 4 g per ring according to a conventional method.

<1-5> Preparation of granules

Mixed extract of the present invention 150 mg

Soybean Extract 50 mg

Glucose 200 mg

Starch 600 mg

After mixing the above components, 100 mg of 30% ethanol was added and dried at 60° C. to form granules, which were then filled into bags.

<Production Example 2> Production of food

<2-1> Manufacture of confectionery and snacks

0.5 to 5.0 parts by weight of the mixed extract of the present invention was added to wheat flour, and bread, cakes, cookies, crackers, and noodles were prepared using the mixture to prepare health-enhancing foods.

<2-2> Manufacture of dairy products

5 to 10 parts by weight of the mixed extract of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<2-3> Manufacture of wire

Brown rice, barley, glutinous rice, and adlay were alphanized by a known method, dried, roasted, and then prepared into a powder having a particle size of 60 mesh using a grinder. Black beans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, roasted, and then prepared into powder with a particle size of 60 mesh by a grinder.

The dried extract obtained by concentrating the mixed extract of the present invention under reduced pressure in a vacuum concentrator and drying with a spray or hot air dryer was pulverized with a grinder to a particle size of 60 mesh to obtain a dry powder. It was prepared by blending the dry powder of grains, seeds and SBE prepared above in the following ratio.

Cereals (brown rice 30 parts by weight, adlay 15 parts by weight, barley 20 parts by weight),

Seeds (7 parts by weight of perilla seeds, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds),

Kkujippong extract of the present invention or its enzymatic hydrolyzate dry powder (3 parts by weight),

Reishi (0.5 parts by weight), and Rehmannia Root (0.5 parts by weight)

<Production Example 3> Production of beverage

1000 mg of mixed extract of the present invention

Citric Acid 1000 mg

100 g of oligosaccharides

2 g plum concentrate

1 g of taurine

Add purified water to make a total of 900 ml

After mixing the above ingredients according to the usual health drink manufacturing method, 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, and then refrigerated. It is used for manufacturing the health food of the invention.

Although the composition ratio is a mixture of ingredients suitable for a relatively favorite beverage in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the class of demand, the country of demand, and the purpose of use.

Claims (16)

A pharmaceutical composition for preventing or treating female menopausal symptoms, comprising Cudrania tricuspidata extract, Angelica sinensis extract, and Lycii Radicis Cortex extract as active ingredients.
The pharmaceutical composition for preventing or treating female menopausal symptoms according to claim 1, wherein the extract is extracted with water, C ₁ to C ₂ lower alcohol or a mixed solvent thereof.
The pharmaceutical composition for preventing or treating female menopausal symptoms according to claim 2, wherein the extract is extracted with one or more solvents selected from the group consisting of water, methanol, ethanol, and mixed solvents thereof.
The pharmaceutical composition for preventing or treating female menopausal symptoms according to claim 1, wherein the extracts of kkujippong and Angelica gigas are at least one extract selected from flowers, branches, stems, leaves, fruits, aerial parts, rhizomes and roots. .
The pharmaceutical composition for the prevention or treatment of female menopausal symptoms according to claim 1, wherein the composition contains kkujippong extract: angelica extract: jigolpi extract in a weight ratio of 1: 1 to 1000: 1 to 500.
The pharmaceutical composition for preventing or treating female menopausal symptoms according to claim 1, wherein the composition contains kkujippong extract: angelica extract: jigolpi extract in a weight ratio of 1:300 to 800:50 to 400.
The method of claim 1, wherein the female menopausal symptoms include hot flashes, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urethral atrophy, vaginitis, cystitis, dysuria, urgency, concentration disorder, short-term memory disorder, anxiety, nervousness, memory A pharmaceutical composition for preventing or treating female menopausal symptoms, characterized in that at least one selected from the group consisting of decline, muscle pain, joint pain and osteoporosis.
The pharmaceutical composition for preventing or treating female menopausal symptoms according to claim 1, wherein the composition is an estrogen receptor agonist.
The pharmaceutical composition for preventing or treating female menopausal symptoms according to claim 1, wherein the composition has estrogenic activity.
A health functional food for preventing or improving female menopausal symptoms, containing cudrania extract, Angelica gigas extract, and Jigolpi extract as active ingredients.
The health functional food for preventing or improving female menopausal symptoms according to claim 10, wherein the extract is extracted with water, C ₁ to C ₂ lower alcohol or a mixed solvent thereof.
The health functional food for preventing or improving female menopausal symptoms according to claim 10, wherein the extracts of kkujippong and Angelica gigas are extracts of one or more selected from flowers, branches, stems, leaves, fruits, aerial parts, rhizomes and roots. .
[Claim 11] The health functional food for preventing or improving female menopausal symptoms according to claim 10, wherein the composition contains kkujippong extract: angelica extract: jigolpi extract in a weight ratio of 1: 1 to 1000: 1 to 500.
The method of claim 10, wherein the female menopausal symptoms include hot flashes, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urethral atrophy, vaginitis, cystitis, dysuria, urgency, concentration disorder, short-term memory disorder, anxiety, nervousness, and memory. Health functional food for preventing or improving female menopausal symptoms, characterized in that at least one selected from the group consisting of decline, muscle pain, joint pain and osteoporosis.
A food composition for preventing or improving female menopausal symptoms, containing kkujippong extract, angelica extract, and gigolpi extract as active ingredients.
The method of claim 15, wherein the female menopausal symptoms include hot flashes, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urethral atrophy, vaginitis, cystitis, dysuria, urgency, concentration disorder, short-term memory disorder, anxiety, nervousness, and memory. A food composition for preventing or improving female menopausal symptoms, characterized in that at least one selected from the group consisting of decline, muscle pain, joint pain and osteoporosis.

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