KR20190113127A - Composition for improving female climacteric symptoms comprising extract of Trifolium pratense L. - Google Patents

Abstract

The present invention relates to a composition for improving women's menopause, and more particularly, to a composition for improving women's menopause containing red clover ( Trifolium pratense L.) extract as an active ingredient.
According to the present invention, the declover ( Trifolium pratense L.) extract has an estrogen-like activity that activates the signaling of ER (Estrogen Receptor), so as a raw material of pharmaceutical compositions or functional foods that can improve the symptoms of female menopausal symptoms Can be used.

Description

Composition for improving female climacteric symptoms comprising extract of Trifolium pratense L.}

The present invention relates to a composition for improving women's menopause, and more particularly, to a composition for improving women's menopause containing red clover ( Trifolium pratense L.) extract as an active ingredient.

Estrogen is a hormone secreted by follicles in the ovary. It develops the genitals and displays its function to develop secondary sexual characteristics and promote uterine development, endometrial growth, mammary gland development, and regular menstruation. Reduced secretion of estrogens affects the urinary and reproductive systems of women as well as other parts of the body. In postmenopausal women, the rapid decrease in estrogen causes symptoms such as hot flushes, sudden heartbeat changes, and sleep disorders, which are the most frequent menopausal symptoms. This lack of estrogen can lead to neurosis, irritability, poor concentration, depressed mood, anxiety, loss of confidence and sexual desire. In addition, due to the reduction of estrogen blood supply is not made smoothly and lose the elasticity and moisture of the skin and vaginal mucosa. Since this causes chronic diseases, the necessity of treatment for postmenopausal women is gradually increasing. As a method for treating symptoms of postmenopausal women, hormone replacement therapy (HRT) which artificially administers estrogen is widely used. have. As a substitute for synthetic estrogens used in hormone replacement therapy, researches are actively conducted to find natural estrogens derived from natural products, especially plant-derived plants, which have similar activities as synthetic estrogens and have fewer side effects. In particular, natural estrogens derived from plants are commonly referred to as phytoestrogen. The phytoestrogens generally refer to nonsteroidal compounds derived from plants having a similar function and structure to female hormone estrogens. Found in flowers or seeds. Examples of the phytoestrogens are largely isoflavone (isoflavone) derived from legumes (쿠), cumumes (coumestan) derived from clover, lignan (lignan) derived from flax seeds. Isoflavones exist in plants in the inactive forms of biochanin A and formononetin, but they are absorbed into the body and are ingested by glucosidase of intestinal bacteria, each of which has an estrogen-like structure. It is converted to genistein and daidzein and shows estrogen-like activity. Genistein has been reported to have anti-cancer effects that inhibit the growth of cancer cells by inhibiting protein tyrosine kinase in cells. (Botanical antioxidants in the prevention of photocarcinogenesis and photoaging.Exp. Dermatol. 2006. 15: 678-684.) However, it may cause cancer in people who are ineffective or have genetic predisposition to uterine and breast cancer. It has also been reported. Therefore, there is a need to develop new phytoestrogens having good estrogen activity without causing side effects such as those of genistein.

Therefore, the present inventors have objectively verified natural resources, which have been used for a long time, and have been tested in human breast tissue-derived cell line, MCF-7 cells, to develop a natural substance that has the effect of treating, improving or preventing women's menopausal syndrome. As a result of searching for a raw material having MCF-7 cell proliferation activity through the Estrogen Receptor (ER), it was confirmed that the composition containing the red clover extract was excellent in preventing, improving and treating female menopausal disease. The invention was completed.

KR 10-1798882 B KR 10-2018-0006477 A

Therefore, the main object of the present invention is to provide a composition for improving women's menopause containing red clover ( Trifolium pratense L.) extract having estrogen-like activity.

Another object of the present invention to provide a pharmaceutical composition or functional food for improving women's menopause containing the composition.

According to one aspect of the invention, the present invention provides a composition for improving women's menopause containing red clover ( Trifolium pratense L.) extract as an active ingredient.

As the woman ages and the ovary ages and the function decreases, ovulation and the production of female hormones no longer occur. This phenomenon is called menopause. It is usually diagnosed as menopause when there is no menstruation for one year. This change usually begins in the mid-to-late 40s and progresses gradually, starting at about 1 year after menopause appears completely absent from menopause, more commonly referred to as menopause, with an average duration of 4-7 years. About a year. During menopause, women experience symptoms such as hot flushes, sudden heartbeat changes, and sleep disorders due to a rapid decrease in estrogen.

In the present invention, the female menopausal symptoms are skin flushing, hot flashes, hot flashes, sweating, numbness, joint pain, muscle pain, reduced skin elasticity, dry skin, fatigue, palpitations, insomnia, depression, anxiety, headache, Nervousness, vaginal powder

It provides a composition for improving women's menopause, characterized in that any one or more of reduced rain, vaginal dryness, vaginal contractility, dyspareunia and sexual dysfunction.

Red clover ( Trifolium pratense L.) is a legume native to Europe and is also called red shamrock. It has been cultivated for a long time, which has been cultivated for animal feed, but it has grown wild. Red clover is also called purple clover because it is dark pink. Familiar with shamrock in Korea, red clover is a herb used in herbal medicine since ancient Roman times. It is widely applied to blood purification, natural hormonal agent, menopausal women's disease, nervous stability and strengthening of immune system. Flowers have anti-inflammatory and diuretic effect, and dried flowers and drinking herbal tea improves eczema and cough. Also used for arthritis.

In the present invention, the extract can be extracted with any solvent conventionally used, preferably characterized in that extracted with water, lower alcohol having 1 to 5 carbon atoms or a mixed solvent thereof. The lower alcohol contains about a week.

In one embodiment of the present invention, the extract may be an ethanol extract, but is not limited thereto. Depending on the organic solvent to be extracted, the degree of extraction and loss of the active ingredient of the medicine may be different, so select an appropriate organic solvent. The extraction method is not particularly limited and includes, for example, hot water extraction, cold extraction, ultrasonic extraction and reflux extraction. When the extract is concentrated, methods such as concentration under reduced pressure and reverse osmosis concentration may be used. The drying step after concentration includes spray drying, hot air drying, freeze drying, vacuum drying, reduced pressure drying, foam drying, high frequency drying, infrared drying, and the like.

In the present invention, the red clover ( Trifolium pratense L.) extract provides a composition for improving women's menopause, characterized in that it has an estrogen-like activity that activates the signaling of ER (Estrogen Receptor). The composition according to the present invention can be used as a raw material for pharmaceutical compositions or functional foods that can improve female menopausal symptoms.

In the present invention, the composition provides a composition for improving women's menopause, characterized in that it is prepared in the form of a powder, granules, tablets, capsules, tea bags or suspensions. The composition may preferably be formulated in the form of a dietary supplement.

The composition of the present invention may be ingested orally in the form of tea, beverage or dietary supplement. At this time, the intake amount of the red clover ( Trifolium pratense L.) extract according to the present invention may vary depending on the degree of need for improving women's menopausal symptoms, it is preferable to intake in an amount of 100 ~ 600mg / kg body weight.

According to the present invention, the red clover ( Trifolium pratense L.) extract has an estrogen-like activity that activates the signaling of ER (Estrogen Receptor), thereby exhibiting an effect on improving menopausal symptoms caused by a decrease in secretion of female hormone estrogen. have.

1 is a graph evaluating the activity of inducing the proliferation of ER-positive breast adenocarcinoma cells (MCF-7) of the red clover extract and dermis extract of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example 1: Preparation of Extract

To compare the effects of plant extracts on female menopausal effects, 100g of dried dermis (mandarin peel) and red clover ( Trifolium pratense L.) were extracted for 3 hours at 80 ° C using 1L of 50% ethanol. Concentrated to 25 brix using (Eyela N-1300). The concentrated sample was prepared by using the Gyeonggi Bio Center freeze dryer (Labconco 79340-10). Each extract powder was dissolved and used at 1 mg / ml for the evaluation of the menopausal improvement effect using the cells.

Example 2: Estrogen-like activity evaluation experiment

The experiment introduced by MCF-7 cell proliferation: Soto et al., 1995 (1) was performed. The cell proliferation of selected extracts was investigated using MCF-7 cell line (KTCC, Seoul, Korea), a cell line derived from human breast cancer.

The MCF-7 cell line is a cell line derived from human breast cancer and is a useful cell line for evaluating the estrogenicity of test substance because it expresses abundantly estrogen receptor. In the MCF-7 cell line, when a substance with estrogenicity acts as a ligand and binds to the estrogen receptor, the conjugate enters the nucleus and acts as a transcription factor, resulting in cell proliferation.

Phenol-red free media (CDFBS-DMEM) containing charcoal dextran treated serum was used to thoroughly exclude external estrogen effects in cell culture and test material treatment. Incubate MCF-7 cells in 96-well plates for 10 ⁴ cells / well for 24 hours, remove the medium, and test medium (dermis, red clover) at a constant concentration (5, 10, 25, 50, 100 ug / ml). Medium was provided. CD-DMEM medium containing the test substance was prepared by diluting a concentration of a test substance stock solution dissolved in DMSO 500-1000 times in the medium immediately before the experiment. In order to confirm that MCF-7 cell proliferation was ER-mediated, the test with and without the ER-antagonist ICI 182,780 was performed. ICI 182,780 was prepared at a concentration of 10 ^-7 M in 1% FCS, 6 ng ml ^-1 insulin, 2.5 mM Glutamax solution, and treated with a concentration of 10 ^-7 M in MCF-7 cells. DMSO (0.5% ~ 0.1%) was added to CDFBS-DMEM and vehicle control group was treated with 1 nM 17β-ES and 1 μM genistein. 48 hours incubation at 37 ℃, 5% CO ₂ incubator, MTT assay was performed to determine the number of live cells. After completely removing the culture solution, MTT [3- (4,5-dimethylthiazol-2-yl-)-2,5-diphenyltetrazolium bromide, Sigma] solution dissolved in DPBS was treated with 100 μl volume at a concentration of 1 mg / ml, followed by 37 ℃, Incubated for 1 hour in a 5% CO ₂ incubator. After incubation, the solution was completely removed, and isopropanol was added to lyse the cells. At the same time, purple MTT crystals produced by living cells were dissolved. The amount of MTT crystals dissolved in isopropanol was measured at 560 nm by ELISA Plate Reader (Glomax, promega). The experimental result is shown in FIG.

Substances with estrogen-like activity can induce proliferation of ER-positive breast adenocarcinoma cells (MCF-7) by activating the signaling of ER (Estrogen Receptor). Appear proportionally). Using this principle, the dermal and red clover extracts were reacted with MCF-7 cells to confirm the effect of improving women's menopausal effects.

In both raw materials, it was confirmed that the cell proliferation effect increased as the treatment concentration was increased at the concentration of 5 ug / ml or more, and the cell proliferation effect of 25 ug / ml of red clover and 100 ug / ml of the dermis was similar. It was confirmed that the red clover showed high estrogen-like activity.

However, when the extract was treated with the ER antagonist ICI 182,780, the cell proliferation effect of MCF-7 was hardly confirmed as compared to the treatment with red clover extract. Therefore, the estrogen-like activity of the red clover extract was ER According to this mechanism, it is expected that red clover extract can be used as a raw material for food for improving menopausal symptoms.

As described above, according to the present invention, the red clover ( Trifolium pratense L.) extract has an estrogen-like activity that activates the signaling of ER (Estrogen Receptor), thereby improving the menopausal symptoms caused by a decrease in the secretion of female hormone estrogen. Can be effective.

Claims (5)

Red clover ( Trifolium pratense L.) A composition for improving women's menopause containing as an active ingredient. According to claim 1, wherein the red clover ( Trifolium pratense L.) extract is a composition for improving women's menopause, characterized in that extracted with water, lower alcohol having 1 to 5 carbon atoms or a mixed solvent thereof. The composition of claim 1, wherein the red clover ( Trifolium pratense L.) extract has an estrogen-like activity that activates signaling of ER (Estrogen Receptor). The composition of claim 1, wherein the composition is a pharmaceutical composition or a functional food composition. The method of claim 1, wherein the composition is a powder, granules, tablets, capsules, tea bags or a composition for improving female climacteric, characterized in that in the form of a suspension.

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