KR20150037101A - Composition for treating or preventing postmenopausal syndrome - Google Patents

Abstract

The present invention relates to a composition for treating or preventing postmenopausal syndrome comprising hypericin as an active ingredient. The hypericin of the present invention promotes expression of estrogen responsive element (ERE) in MCF-7 cells which are human breast tissue derived cells and, more specifically, expression of estrogen receptor (ER), and promotes growth of the MCF-7, thereby having estrogen activity. Also, the hypericin has enhanced stability by not having cytotoxicity; and is originated from plants that have been drunk or eaten for long time, thereby having no side effects. Therefore, the hypericin which is the active ingredient of the composition of the present invention can be usefully applied in treating or preventing postmenopausal syndrome.

Description

Technical Field [0001] The present invention relates to a composition for treating or preventing female menopausal syndrome,

The present invention relates to a composition for the treatment, improvement or prevention of female postmenopausal syndrome comprising a composition for treatment, improvement or prevention of female postmenopausal syndrome, specifically a component derived from a natural product.

Women go through a series of sexual maturation as they get older. This period is called puberty. This refers to a process in which an individual's physical structure and emotions reach a state where they can fulfill their original functions. After puberty, women secrete estrogen and progesterone, the female hormones. Specifically, when the gonadotropin secreted by the pituitary gland transfers a signal, it regulates the menstrual cycle and releases hormones such as Folliocle Stimulating Hormone (FSH) and Lutenizing Hormone (LH) ), And the follicle-stimulating hormone and the luteinizing hormone act on the ovary again to secrete estrogen and progesterone.

The estrogen is a hormone secreted from the follicles in the ovary. It promotes the development of the reproductive organs and functions to develop secondary sex and promotes uterine growth, endometrial hyperplasia, development of the mammary gland, regular menstruation, and the like. In addition to the ovaries, the estrogen is secreted in a small amount from the adrenal cortex and testes, including the placenta. Three kinds of steroids such as estrone, estradiol and estriol found in the human body are known.

Among the above estrogens, 17 beta-estradiol (E2), the most abundant and strongest estrogen in women before menopause, is synthesized while producing follicles and secreted into the blood to partially bind sex hormone-binding globulin and flow to cells throughout the body . The main pathway of the estradiol metabolism is that the estradiol is reversibly oxidized to estrone (E1) which is a weak estrogen and then converted to estriol (E3).

The estrogen stimulates the growth of endometrium, uterine muscle, vagina and urethral epithelium, smoothes vascular flow in the reproductive tract, increases secretion of uterine glands, and induces progesterone and luteinizing hormone receptor expression. In addition to the development of female reproductive organs and secondary manifestations, the estrogen affects skeletal growth and development, female fat distribution and lipid metabolism, and also acts on skin and collagen tissues, neurons and cardiovascular system.

In women, the function of the ovaries declines with age, and this leads to a significant decrease in estrogen secretion. Due to the reduced secretion of estrogen, the response to follicle-stimulating hormone and luteinizing hormone is reduced, the initial follicle shortening is shortened, the menstrual cycle is further shortened, the ovulation phase is further shortened, and progesterone production is reduced. Eventually, the follicle becomes unresponsive and does not produce estrogen.

It is known that climacteric is a transitional period in which physiological function and sexual function are reduced or eliminated due to an overall and gradual decrease of ovarian function, and ovulation is stopped after ovarian function is stopped as a part of menopausal period, A permanent stop is called menopause. On the other hand, the World Health Organization (WHO) defined menopause as a period when ovarian function is lost and estrogen secretion is no longer conceived, and that the transition from adulthood to old age is over.

As described above, women gradually start ovarian function gradually from the age of 40 after the menopausal period, and the average life span of women is gradually extended, the period of hormone deficiency is about 1/3 to half of the life span. Thus, there is an increasing need for special attention to the healthy life of women after menopause.

For example, in Korea, the proportion of elderly people aged 65 or older is estimated to be 9.1% in 2005, and it is expected to be more than 14% of the population in 2019. The average life expectancy of women is higher than that of men In 2002, the average life span of women was 80.4 years. Therefore, women in Korea should live for about 40 years after menopause. In this respect, interest in women's health after menopause is increasing .

The chronic chronological symptoms due to changes in the body during the menopausal period, including the menopause or previous menstrual changes due to changes in hormones, specifically estrogen production, follicle stimulating hormone and luteinizing hormone, are called menopausal symptoms or menopausal symptoms.

In particular, the decrease in estrogen affects not only the female urinary system, genitalia but also other parts of the body, so estrogen secretion decreases greatly throughout the body.

For example, most menopausal women are experiencing acute female hormone deficiency symptoms such as hot flush, tachycardia, sweating, anxiety or sleep disorder, and in some women, chronic female hormone deficiency causes cardiovascular disease, osteoporosis Of chronic illnesses.

Specifically, symptoms of menopausal symptoms include symptoms due to vascular changes such as hot flush, night sweat, tachycardia or headache, and symptoms due to musculoskeletal changes such as myalgia, joint pain and back pain , Memory loss, depression, loss of concentration, anxiety, and dizziness.

In addition, since the estrogen is reduced, the supply of blood is not smoothly performed and the elasticity and moisture of the skin and vaginal mucosa are lost. For example, when the mucous membranes of the eyes, nose, or mouth are dry, they may experience uncomfortable eyes or thirsty burns.

In addition, symptoms such as vaginal dryness, urinary tract infections, urinary frequency or urinary incontinence appearing in the years after menopause and epidermal symptoms, osteoporosis after years of menopause, cardiovascular disease such as atherosclerosis or cerebrovascular disease . In particular, heart disease is one of the major causes of death in postmenopausal women, and many women are suffering from osteoporosis because of the rapid bone loss caused by estrogen deprivation after menopause.

As described above, the necessity of treatment for postmenopausal women is becoming more and more important, and various methods have been studied for the treatment of the symptoms of postmenopausal women, namely, postmenopausal syndrome. For the treatment of postmenopausal syndromes, Hormone Replacement Therapy (HRT) is widely used to administer estrogen.

There are two types of estrogen: natural estrogen and synthetic estrogen. Oral administration is the most widely used method of administering the synthetic estrogen hormone preparation, and parenteral administration includes transdermal patches, transdermal cream, gel, and vaginal cream.

However, such an artificial synthetic estrogen is effective for the improvement of the menopausal symptoms, but it increases the activity of the cancer-inducing gene in the long-term use, thereby posing a risk of uterine cancer or breast cancer and possibly causing venous thrombosis or gallbladder disease. Is being heated.

Accordingly, studies have been made actively to find a estrogen derived from a natural product having a similar activity to a synthetic estrogen as a substitute for the synthetic estrogen used in the above-mentioned hormone replacement therapy, while having a less side effect.

Premarin, made from natural product-derived estrogens and from the animal's natural urine, has been shown to increase the levels of estradiol and estrone in clinical trials to a monthly level. However, there have been reports of serious side effects such as increased breast cancer (26%), heart attack (29%) and fainting (44%), and blood coagulation (2 times) It ended itself in five years, and the US Food and Drug Administration (FDA) decided in 2003 to put a warning on hormone medicines.

Although the use of conventional hormone replacement agents such as synthetic estrogen and animal estrogen products can cope with overall menopausal symptoms, there is a problem that their use is limited due to serious side effects.

Therefore, there is a trend toward research on natural estrogen derived from plant having a similar activity to synthetic estrogen as a substitute for synthetic estrogen used in the above-mentioned hormone replacement therapy and having less side effects.

In particular, plant-derived natural estrogens are often referred to as phytoestrogens.

The vegetable estrogen refers to a non-steroidal compound derived from a plant which is generally similar in function and structure to female hormone estrogen, and is found in the stem, root, flower or seed of a plant.

Examples of the above vegetable estrogens include isoflavone derived from soybean and bean, coumestan derived from clover, and lignan derived from flax seed. In addition, there are flavanones, flavones, dihydrocalchones, and the like.

For example, isoflavones exist as inactive glycosides of biochanin A and formononetin in plants, but are absorbed into the body and are similar to estrogen by glucosidase in intestinal bacteria (Genistein) and daidzein (daidzein), resulting in estrogen-like activity.

Genistein, which is a representative example of the vegetable estrogen, has been reported to inhibit the protein tyrosine kinase of cells and thus to inhibit the growth of cancer cells. However, it has also been reported that it may cause cancer in people who have no effect or who have a genetic predisposition for uterine cancer and breast cancer.

Therefore, it is required to develop new phytoestrogenic estrogens having excellent estrogenic activity without side effects such as the side effects of genistein. Due to this necessity, it is urgently required to objectively verify the natural resources that have been used for a long time as a food and have proved its safety, and to develop a natural substance having a therapeutic, improving or preventive effect on female postmenopausal syndrome.

KR 10-1067028 B KR 10-1075554 B KR 10-2007-0116981 A KR 10-2001-0071822 A

It is an object of the present invention to provide a composition for treating, ameliorating or preventing female menopausal syndrome which comprises a natural product-derived component effective for the treatment, prevention or amelioration of female menopausal syndrome as an active ingredient. .

In order to achieve the above object, the present invention provides a composition for treating or preventing female menopausal syndrome. The composition for treating or preventing female menopausal syndrome may be a pharmaceutical composition or a pharmaceutical composition.

More specifically, the present invention provides a pharmaceutical composition for the treatment or prevention of female postmenopausal syndrome comprising hypericin as an active ingredient.

The hyperlysine may be selected from the group consisting of Hypericum perforatum extract, in particular extracts of flower and leaf blends of Leucocephala. The extract can be a solvent extract prepared by a solvent extraction method, specifically, a solvent extract extracted with water, an alcohol having 1 to 5 carbon atoms, or a mixture thereof.

The female menopausal syndrome may be any one selected from the group consisting of facial flushing, osteoporosis, and venous thrombosis.

The present invention also provides a food composition for improving or preventing female menopausal symptoms comprising hypericin as an active ingredient.

The hyperlysine may be selected from the group consisting of Hypericum perforatum extract, in particular extracts of flower and leaf blends of Leucocephala. The extract can be a solvent extract prepared by a solvent extraction method, specifically, a solvent extract extracted with water, an alcohol having 1 to 5 carbon atoms, or a mixture thereof.

The female menopausal syndrome may be any one selected from the group consisting of facial flushing, osteoporosis, and venous thrombosis.

The food composition for improving or preventing female menopause syndrome may be a health functional food for improving or preventing female menopausal symptoms.

The inventors of the present invention have long been used as foods for the development of phytoestrogens which exhibit excellent estrogenic activity but do not cause side effects such as synthetic estrogens or animal estrogens used in conventional hormone replacement agents and toxicity In the study of its safety-proven natural resources, the extracts of Jojang extract, specifically extracts from leaves and leaves of Junjangjang, were extracted from MCF-7 cells, the human breast tissue-derived cell line, and the estrogen responsive element ERE) specifically promotes the expression of the estrogen receptor (ER) and promotes the growth of MCF-7, and thus has an effect on estrogenic activity, and it has been confirmed that estrogen receptor In studying the ingredients, Hypericin and hyperforin were identified during the analysis of the pharmacological components contained in the extracts and fractions having excellent estrogenic activity among the herbal extracts and the fractions of the extracts of the herbal extracts. Confirming that hyperlysine and hyperferrin have excellent estrogenic activity, and completed the present invention.

As used herein, unless otherwise specified, the active ingredient refers to a component that provides a therapeutic, ameliorative, or preventive effect, particularly an estrogen-like or estrogen-like activity, in a composition, particularly a pharmaceutical composition or a food composition, of a female postmenopausal syndrome do.

Unless otherwise specified herein, hyperlysine is a compound having the general formula C ₃ O H ₁₆ O _{5 with} a molecular weight of 504.44, specifically 1,3,4,6,8,13-hexahydroxy-10, (1, 10, 9, 8, opqra) perylene-7,14-dione, more specifically a compound having the structure of the following formula (1): Hypericum sp. It is a natural pigment separated from plants.

[Chemical Formula 1]

Unless otherwise specified herein, a derivative refers to a compound that can be modified by modifications that are generally recognized by those skilled in the art, including, for example, esters of acids, amides of acids , A protecting group such as a benzyl group for an alcohol or a thiol, and the like.

As used herein, the term " extract " means an extract obtained by treating an extractive solvent with a plant or a product prepared by the method of producing an extract, unless otherwise specified. The extract may be formulated ) Results.

The extract has the meaning of being used as a crude extract, but broadly includes fractions obtained by further fractionating the extract. Specifically, the above-mentioned extract includes not only an extract obtained by using an extraction solvent, but also an extract obtained by additionally applying a purification process thereto. For example, the extract may contain a fraction obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, a fraction obtained by various chromatographies (prepared for separation according to size, charge, hydrophobicity, or affinity) Further comprising fractions obtained through various purification methods.

Unless otherwise specified herein, a pharmaceutically effective amount means an amount sufficient to exert pharmacological efficacy.

Unless otherwise specified herein, estrogen is a female hormone that is mainly secreted in the follicles and progesterone in the ovary of an animal, and is also secreted in the placenta and affects the reproductive cycle. The estrogen is a steroid hormone with a benzene ring and is chemically very stable. Three kinds of estrone (E1), estradiol (E2) and estriol (E3) are well known. Since estrogen directly affects the reproductive cycle, it can be used as a treatment for abnormalities in menstruation, in contraceptive pills, and because menopause or menopausal syndrome is caused by estrogen deficiency when women become menopausal, It may also be used in hormone therapy for the treatment or prevention of disorders or postmenopausal syndromes.

As used herein, unless specified otherwise, menopausal syndrome or menopausal symptoms refers to symptoms resulting from a decrease in hormone production in postmenopausal women, and generally includes facial flushing, sweating, sweating during sleep, dry skin, vaginal dryness, vaginal dryness Is a concept that includes atrophy, atrophy of the lower urinary tract, intercourse, vaginitis, cystitis, dysuria, urinary incontinence, concentration disorders, short term memory impairment, anxiety, nervousness, memory loss, loss of libido, myalgia, arthralgia or osteoporosis.

Unless otherwise specified herein, treatment or amelioration refers to any action by which administration of a composition of the invention improves or alleviates symptoms of female postmenopausal syndrome.

As used herein, " prophylactic " means any action that inhibits the symptoms of female postmenopausal syndrome by administration of the composition of the present invention, unless otherwise specified herein.

In this specification, unless otherwise indicated, administration means to provide any desired composition of the invention to the subject by any suitable method.

As used herein, unless otherwise specified, an individual refers to any animal, particularly a mammal, including humans, monkeys, dogs, goats, pigs or rats, who have undergone female polyneuropathy syndrome by administration of the composition of the present invention.

Unless otherwise specified herein, a pharmaceutically effective amount means an amount sufficient to treat a female menopausal syndrome at a reasonable benefit or risk ratio applicable to medical treatment, including the type, severity, severity of a female postmenopausal syndrome, The activity of the compound, the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, the factors including the drugs used simultaneously and other factors well known in the medical arts.

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

The present invention relates to a pharmaceutical composition for the treatment or prevention of female postmenopausal syndrome comprising hypericin as an active ingredient. The hyperlysine may be isolated from the extract of A. chinensis.

The Hypericum perforatum , St. John? wort) is one of the herbs originating in Europe and West Asia. It grows well in the sunlight as well as in the half-sword, so it is easy to grow at home, and it breeds by bending and sowing. It has been known from the past that it has been eaten as a herbal tea for the purpose of treating depression, treating insomnia, relieving stress, etc., and has been known to have effects such as astringent action, antibacterial action or anti- .

Preferably, the extract is a mixture of flowers and leaves of at least one selected from the group consisting of flowers and leaves, especially flowers and leaves, and the like. The extraction solvent may be at least one selected from the group consisting of water, an alcohol having 1 to 5 carbon atoms, diluted water of the alcohol, and a mixture thereof.

The extract may be a fraction prepared by extracting with an extraction solvent or extracting with an extraction solvent and fractionating the extract with a fraction solvent. The fraction is extracted with an extraction solvent and extracted with any one solvent selected from the group consisting of hexane, methylene chloride, acetone, ethyl acetate, ethyl ether, chloroform, water and mixtures thereof, preferably hexane, methylene chloride and ethyl acetate , More preferably at least one solvent selected from the group consisting of ethyl acetate, and more preferably ethyl acetate.

As described above, the recombinant vector pERα-RL-SV40 (pERα-RL-SV40 plasmid) and the pERβ-RL-SV40 recombinant vector (pERβ-RL-SV40 plasmid) The luciferase activity was measured in the transfected MCF-7 cell line (human breast cancer cell line) to confirm the estrogen receptor (ER) activation degree. As a result, excellent luciferase activity was confirmed, and the MCF-7 cell line As a result, it was confirmed that the hyperlysine could treat, ameliorate or prevent the menopausal syndrome (see FIG. 3 and FIG. 4). Particularly, among the fractions of the aqueous extracts of 80% ethanol aqueous solution, the ethyl acetate fraction was found to promote the growth of the MCF-7 cell line. As a result, the MCF-7 cell line growth was compared with the fractions fractionated by other fraction solvents, . As a result of repeatedly checking the active ingredient of the fraction, it was confirmed that the active ingredient of the fraction was hyperlysine.

In addition, since hyperlysine is isolated from a natural plant, Johansoch, it is similar to a processed product produced by processing a natural product, which is a natural product that has been used for a long time as a food ingredient of food or a beverage such as tea, , And it was confirmed that the cytotoxicity was not a problem.

In this regard, the hyperlysine may be used for the treatment or prevention of female postmenopausal syndrome. From this viewpoint, the hyperlysine can be used as an active ingredient of a pharmaceutical composition for treating or preventing female menopausal syndrome of an extract.

The composition for treating or preventing female menopausal symptoms comprising hyperlysine as an active ingredient can be directly applied to animals including humans.

The term postenopausal syndrome refers to all symptoms caused by a decrease in estrogen not only in menopause but also in perimenopausal and postmenopausal periods.

The full-blown competition is a period before the actual menopause starts. During the entire menopausal period, the generation of estrogen is irregular, and the fertility of the woman is greatly reduced. The full-closed event may last for several months or several years, and climacteric symptoms may occur in the full-closed event.

The menopausal syndrome may be selected from the group consisting of sexual intercourse, vaginal dryness, sleep disorder, mental awareness problem, arthralgia, musculoskeletal disease, dyspepsia, urinary incontinence, fatigue, Climacteric symptoms such as night sweats, palpitations, depression, anxiety and irritability, and further include severe diseases or conditions caused by estrogen deficiency, such as osteoporosis, Genitourinary system disease, cardiovascular disease, dementia, tooth loss or skin collagen loss, and the like.

The female menopausal syndrome is preferably any one selected from the group consisting of facial flushing, osteoporosis, and venous thrombosis.

The composition for treating or preventing female menopausal symptoms comprising hyperlysine as an active ingredient may contain 0.00001% to 50% by weight of the hyperlysine based on the total weight of the composition, but is not limited thereto.

The composition for treating or preventing female menopausal symptoms comprising hyperlysine as an active ingredient may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of medicaments.

The composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method have.

Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

When the composition of the present invention is formulated, it may be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used.

Solid formulations for oral administration of the compositions of the present invention include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium Calcium carbonate, sucrose or lactose, gelatin, and the like. In addition to the above excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration of the composition of the present invention include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, Preservatives, and the like.

Formulations for parenteral administration of the composition of the present invention may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the agent for parenteral administration include non-aqueous solvents, suspensions such as propylene glycol, vegetable oils such as polyethylene glycol, olive oil, injectable esters such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The composition for treating or preventing female menopausal symptoms comprising hyperlysine as an active ingredient may further comprise, as an active ingredient, the hyperlysine alone or a substance having a therapeutic or preventive effect on female menopausal syndrome in the hyperlysine .

Specifically, the composition for treating or preventing female menopausal symptoms comprising the hyperlysine of the present invention as an active ingredient may further comprise, in addition to the above-mentioned effective ingredient, a compound having a therapeutic or preventive effect for a known female postmenopausal syndrome or an extract for a natural product have.

More specifically, the pharmaceutical composition for the treatment or prevention of female menopausal syndrome may further include estrogen in addition to hyperlysine. The estrogen may be estradiol.

The growth of MCF-7 cells, which is a human breast cancer cell line, is inhibited under hormone-deficient culture conditions. When the estrogen component is added under these conditions, the growth of MCF-7 cells is actively increased, , We observed indirectly the estrogenic effect of certain components by observing the growth of MCF-7 cells after treatment with specific components.

The composition for treating or preventing female menopausal symptoms comprising hyperlysine as an active ingredient may further comprise, in addition to the above-mentioned effective ingredients, additional components such as pharmaceutically acceptable or nutritionally acceptable carriers, excipients, diluents Or a subcomponent.

More specifically, the composition for treating or preventing female menopausal symptoms comprising hyperlysine as an active ingredient may further comprise at least one selected from the group consisting of a nutrient, a vitamin, an electrolyte, a flavoring agent, a colorant, a heavy stabilizer, a pectic acid and its salt, A salt thereof, an organic acid, a protective colloid thickener, a pH adjuster, a stabilizer, a preservative, a glycerin, an alcohol, and a carbonating agent used in a carbonated beverage. The carrier, excipient or diluent may be selected from lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acicia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline In the group consisting of celluloses, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, dextrin, calcium carbonate, propylene glycol, liquid paraffin, But the present invention is not limited thereto. Any conventional carrier, excipient or diluent may be used. The components may be added to the active ingredient, i.e., hyperlysine, either independently or in combination.

The composition for treating or preventing female menopausal symptoms comprising hyperlysine as an active ingredient may contain 0.000001% by weight to 50% by weight, preferably 0.00001% by weight to 30% by weight, of the active ingredient, specifically hyperlysine, By weight, more preferably from 0.0001% by weight to 20% by weight.

When the composition for treating or preventing female menopausal symptoms comprising hyperlysine as an active ingredient is weakened, it may be formulated with conventional fillers, extenders, binders, disintegrants, surfactants, anti-coagulants, lubricants, wetting agents, Preservatives, and the like, and they can be used either orally or parenterally.

The formulation of a composition for treating or preventing a female postmenopausal syndrome comprising hyperlysin as an active ingredient may be in a desired form depending on the method of use, and may be used to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal By a method known in the art. Examples of the specific formulations include granules, powders, syrups, liquids, suspensions, tablets, injections, main tablets, cataplasma, capsules, soft or hard gelatin capsules and the like.

Furthermore, compositions for the treatment or prevention of female menopausal syndrome comprising hyperlysine as an active ingredient can be prepared using appropriate methods known in the art or may be prepared using the methods described in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA ). ≪ / RTI >

The dosage of the composition for the treatment or prevention of female menopausal symptoms comprising hyperlysine according to the present invention as an active ingredient may vary depending on factors such as the formulation method, administration method, age, sex and weight of the recipient, severity of disease, , The excretion rate and the responsiveness of the reaction, and the like. As an example, the composition for treating or preventing female menopausal symptoms comprising hyperlysine of the present invention as an active ingredient may be used at a daily dose of 0.000001 mg / kg to 1000 mg / kg or 0.0001 mg / kg / day, 100 mg / kg or 0.003 mg / kg to 50 mg / kg. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The composition for improving or preventing female menopausal symptoms according to another embodiment of the present invention comprises hyperlysine as an active ingredient. The hyperlysine has not only been confirmed to be excellent in estrogenic activity but also has been used for a long time as a food material of food and has no problem in stability and has been found to be free from cytotoxicity as a result of experiments. Since the hyperlysine isolated from the supernatant extract has been identified as a component showing the estrogenic activity of the extract, the hyperlysine can be used for the improvement or prevention of female postmenopausal syndrome. From this viewpoint, the hyperlysine can be used as an active ingredient of a food composition for improving or preventing female menopausal syndrome of an extract.

The composition for the improvement or prevention of female menopausal syndrome includes hypericin as an active ingredient.

As described above, the MCF-7 cell line transfected with the pERα-RL-SV40 recombinant vector (pERα-RL-SV40 plasmid) and the pERβ-RL-SV40 recombinant vector (pERβ-RL-SV40 plasmid) (Human breast cancer cell line), the degree of activation of the estrogen receptor (ER) was confirmed. As a result, the extract of the mixture of the leaves and leaves of the leaves showed remarkably increased luciferase activity in a concentration-dependent manner, And leaves were found to be excellent in the therapeutic activity of the menopausal syndrome (see FIGS. 3 and 4). Particularly, among the fractions of the aqueous extracts of 80% ethanol aqueous solution, the ethyl acetate fraction was found to promote the growth of the MCF-7 cell line. As a result, the MCF-7 cell line growth was compared with the fractions fractionated by other fraction solvents, . As a result of repeatedly checking the active ingredient of the fraction, it was confirmed that the active ingredient of the fraction was hyperlysine. From this viewpoint, the hyperlysine can be evaluated as having excellent effects of improving or preventing female menopausal syndrome.

The menopausal syndrome may be selected from the group consisting of sexual intercourse, vaginal dryness, sleep disorder, mental awareness problem, arthralgia, musculoskeletal disease, dyspepsia, urinary incontinence, fatigue, Climacteric symptoms such as night sweats, palpitations, depression, anxiety and irritability, and further include severe diseases or conditions caused by estrogen deficiency, such as osteoporosis, Genitourinary system disease, cardiovascular disease, dementia, tooth loss or skin collagen loss, and the like.

The female menopausal syndrome is preferably any one selected from the group consisting of facial flushing, osteoporosis, and venous thrombosis.

The composition for improving or preventing female menopausal symptoms comprising hyperlysine as an active ingredient may be a composition containing the hyperlysine alone as an effective ingredient or a substance having an effect of improving or preventing female menopause syndrome in the hyperlysine, A compound having an effect of improving or preventing a known female postmenopausal syndrome or an extract for a natural product.

More specifically, the food composition for improving or preventing the female postmenopausal syndrome may further include estrogen in addition to hyperlysine. The estrogen may be estradiol.

In addition, the food composition for improving or preventing female menopause syndrome may be a health functional food for improving or preventing female menopausal symptoms.

The term " food " means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be eaten directly through a certain degree of processing, Beverages, food additives, beverage additives, and the like.

Such foods include, for example, various foods, beverages, gums, tea, vitamin complexes, and health functional foods. In addition, in the present invention, the food may contain special nutritional foods (e.g., crude oil, spirits, infant food, etc.), meat products, fish products, tofu, jelly, noodles (Such as soy sauce, soybean paste, hot pepper paste, mixed sauce), sauces, confectionery (eg snacks), dairy products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods (E.g., fruit, vegetable beverages, two-oil beverages, fermented beverages, etc.), seasonings (e.g., ramen soup, etc.). The food, the health functional food, the beverage, the food additive and the beverage additive can be produced by a usual production method.

The above-mentioned health functional food refers to a food group to which the function of the food is added to a specific purpose by physical, biochemical, and biotechnological techniques, and to control the bio-defense rhythm of the food composition, Means a food which has been designed and manufactured so that the weight control function related to the living body is sufficiently expressed to the living body.

The health functional food may include food-acceptable food supplementary additives, and may further include suitable carriers, excipients and diluents conventionally used in the production of health functional foods.

The drink refers to a generic term for eliminating thirst or drinking to enjoy a taste, and is intended to include a health functional drink. The beverage is not particularly limited as long as it contains the hyperlysine as an active ingredient in an indicated ratio as an essential ingredient, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages .

Examples of such natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and Xylitol, sorbitol, and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate may be generally about 1 to 20 g, or 5 to 12 g per 100 ml of the food composition of the present invention. In addition, the composition of the present invention may be used for the production of natural fruit juice, fruit juice drink, And may further contain flesh.

In addition to the above, the food composition of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. These components can be used independently or in combination. The proportion of such an additive is not critical, but may be selected from the range of 0 to 200,000 parts by weight per 100 parts by weight of the hyperlysine of the present invention, but is not limited thereto.

The health-functional beverage is used to control the bio-defense rhythm of the beverage group or beverage composition to which the added value is imparted so that the function of the beverage functions for a specific purpose by physical, biochemical, biotechnological, or the like, Means a beverage which has been designed so as to sufficiently express the body controlling function related to the living body.

The health functional beverage is not particularly limited to the other ingredients except that the hypericin of the present invention is contained as an essential ingredient at the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages.

Examples of such natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose, sucrose and the like and polysaccharides such as dextrins, cyclodextrins and the like, and xylitol , Sorbitol, and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably 5 to 12 g per 100 ml of the composition of the present invention.

In addition, in the food composition for improving or preventing female menopausal symptoms comprising hyperlysine as an active ingredient, the effective ingredient may be contained in an amount of 0.0000001 to 10% by weight of the total food, And may be contained in a proportion of 0.00000002 g to 5 g.

The hyperlysine of the present invention was isolated from the extract of Jojang extract. From the results of the experiment, the extract of Jojang extract was found to have estrogenic activity. Further, the extracts of the jujitsu extract and fractions were further tested. As a result, Since hyperlissin has been identified as an active ingredient exhibiting estrogenic activity of the extract, the hyperlysine has been used as a hormone replacement therapy (hereinafter referred to as " hormone replacement therapy ") to prevent, alleviate or treat menopausal syndromes such as facial flushing, osteoporosis and venous thrombosis caused by estrogen deficiency Hormone replacement therapy (HRT), and it is not cytotoxic unlike the conventional therapeutic agent for postmenopausal syndrome, and the hyperlysine is derived from a herb that has been used as a tea or food for a long time. Less likely to occur Since a qualified guess material, and has a superior effect compared with the conventional therapeutic agents for female menopause syndrome.

FIG. 1 is a view illustrating a process for preparing extract and fraction according to an embodiment of the present invention.
FIG. 2 is a graph showing cytotoxicity of an extract of a mixture of L. japonica flowers and leaves according to an embodiment of the present invention. In the graph, CNTs in the horizontal axis represent a control group treated with DMSO as a solvent, The vertical axis represents the relative number of cells measured on the basis of the control group.
FIG. 3 and FIG. 4 are graphs showing estrogenic activity of extracts of a mixture of L. japonica flowers and leaves according to an embodiment of the present invention. FIG. 2 is a graph showing the activity of luciferase activity measured after transfection of ERα into MCF-7 FIG. 3 shows the result of measuring luciferase activity after transfection of ERβ into MCF-7. In each graph, CNTs in the abscissa indicate the negative control group treated with DMSO as a solvent, and positive indicates 17β-estradiol The number on the horizontal axis represents the dose of the extract and the vertical axis represents the degree of fluorescence measured on the basis of the negative control, that is, the luciferase activity.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

< Example 1 . John  The extract and Fraction  Manufacturing>

In the case of Johannesburg, imported Chinese cabbage was used, washed and dried again. The leaves were divided into leaves (flower and leaf mixture). After extracting each part of the extract, 10 g of the supernatant and 200 ml of a 70% aqueous solution of ethanol were placed in an extraction container and extraction was performed at 50 ° C. After the hot water extraction, filtration and concentration were carried out and the extract was dried at -20 ° C.

Further, as shown in Fig. 1, the lyophilized extract prepared by the above-mentioned production method was fractionated while sequentially treating an organic solvent to prepare fractions. Specifically, the lyophilized Prunus mume extract was dissolved in a solvent, and the same amount of hexane was added thereto. The mixture was shaken to the left and right, and left to separate the layers. After separation into a hexane layer and a water layer, the hexane fraction was transferred to water and an equal amount of chloroform was added to the remaining water layer. The solution was shaken to the left and right to allow the two solutions to react well, and then left to separate the layers. When the chloroform layer and the water layer were separated, the chloroform fraction was transferred to water and an equal amount of ethyl acetate was added to the remaining water layer. The solution was shaken to the left and right to allow the two solutions to react well, and then left to separate the layers. Ethyl acetate layer and water layer were separated. The ethyl acetate fraction was transferred to hand and the same amount of butanol was added to the remaining water layer. The solution was shaken to the left and right to allow the two solutions to react well, and then left to separate the layers. The Buthanol layer and the water layer were separated and the butanol fraction was transferred to hand and the remaining water layer was transferred to the hand. Fractions were prepared by evaporating fraction solvents such as hexane, chloroform, ethyl acetate, butanol and water from each of the fractions collected in the manual, followed by drying at -20 ° C.

The prepared extracts and fractions were stored under refrigeration conditions and dissolved in DMSO if necessary.

< Example 2 . John  Determination of Estrogenic Activity of Extracts>

In order to measure the estrogenic activity of the thus-prepared jujuba extract, a sample was first prepared. 17β-estradiol (E8875-1G, Sigma, USA) was prepared with ethanol at a concentration of 3 mM (0.81714 g / ml) and diluted by adding solvent thereto. The extract of Jojang extract was prepared by dissolving the extract of the mixture of Jojoba ganoderma and leaves prepared in Example 1 in DMSO at a concentration of 20 mg / ml and diluting it.

Example  2-1. Toxicity assessment

MCF-7 cells were cultured in a culture medium (DMEM / F12 (11330-032, GIBCO, USA) + 10% FBS (26140-111, GIBCO, USA) + ATCC (American Type of Cell Culture) 1% Penicillin Streptomycin (15140-122, GIBCO, USA) + 1% Bovine insulin (Cellapplications, USA)). After the incubation, the cells were seeded in 96-well plates at a concentration of 2 × 10 ⁴ cells / 100 μl, and then cultured in a 5% CO ₂ incubator at 37 ° C. for 24 hours. After 24 hours of treatment, CCK-8 (ck04-13, Woongbee, Korea) was treated with 10 μl of each extract and the absorbance at 450 nm was measured to confirm cytotoxicity. The absorbance was measured on the basis of a control (CNT) treated with only the same amount of DMSO instead of the sample, and the result is shown in FIG.

As shown in FIG. 2, it was confirmed that the extracts of the mixture of A. chinensis and leaf did not have cytotoxicity at all concentrations.

Example  2-2. Transfected MCF Evaluation of estrogenic activity using -7 cell line

The MCF-7 cell line subcultured in the medium as described in Example 2-1 was seeded into each well at 2 × 10 ⁴ cells / 100 μl per well in a 96-well plate and cultured at 37 ° C. in 5% CO 2 ₂ incubator for 24 hours.

0.11 μg of transfection DNA prepared by injecting ER α (28230, addgene Co., USA) or ER β (35562, addgene Co., USA) into pRL-SV40 vector and 5.2 μl of total reaction solution were mixed with FuGENE HD E3311, Promega, USA) and incubated at room temperature for 10 min. 5 μl of the reaction mixture was added to each well and incubated for 24 hrs. The MCF-7 cell line was incubated with each estrogen receptor (ER ). &Lt; / RTI &gt;

The transfected cells were washed with PBS, and 100 μl of the medium containing the extracts and fractions in the specified amount was added thereto, followed by culturing for 48 hours. After the cultivation, 20 μl of 1 × Passive Lysis Buffer was added to the wells in which the Luciferase Assay (E1501, Promega, USA) was run, and 1 × Renilla Luciferase Lysis Buffer 20 μl was added, and the mixture was stirred for 15 minutes. The lysate dissolved in each of the above lysis buffers was transferred to a white plate (136101, nunc, USA) in a volume of 20 μl, and the degree of fluorescence was measured using a luminometer. At this time, only negative amounts of DMSO were treated with negative control (CNT) and treated with 10 nM of 17β-estradiol in case of positive control. The results are shown in FIG. 3 and FIG.

As shown in FIG. 3, in the case of ERα, the fluorescence activity was increased in a concentration-dependent manner in the case of the extract of the mixture of Isocholate and leaf, and specifically, when the extract of the mixture of Isoflavone and leaf required for DMSO was treated at 60 ppm And 10 nM of a positive control estrogen. When the highest concentration of 100 ppm was used, about 250% of the fluorescence activity was exhibited compared with the negative control. In addition, as shown in FIG. 3, the fluorescence activity was also found in the extract of the mixture of the leaves and the leaves of ERβ. Specifically, when the extract of the mixture of the flowers and leaves required for DMSO was treated at 20 ppm , A remarkably superior fluorescence activity of about 300% to 500% was observed for both positive control as well as positive control.

From the above results, it was confirmed that the extracts of A. chinensis and leaves had excellent estrogenic activity.

Claims (7)

A pharmaceutical composition for the treatment or prevention of female postmenopausal syndrome comprising hypericin as an active ingredient. The method according to claim 1,
Wherein the hypericin is isolated from a solvent extract of a flower and leaf mixture of Hypericum perforatum . The method according to claim 1,
Wherein the female menopausal syndrome is any one selected from the group consisting of facial flushing, osteoporosis, and venous thrombosis. 4. The method according to any one of claims 1 to 3,
Wherein the pharmaceutical composition for the treatment or prevention of female menopausal syndrome further comprises estrogen. A food composition for improving or preventing a female postmenopausal syndrome comprising hypericin as an active ingredient. 6. The method of claim 5,
Wherein the food composition for improving or preventing female menopause syndrome further comprises estrogen. The method according to claim 6,
The food composition for improving or preventing female menopause syndrome is a health functional food for improving or preventing female postmenopausal syndrome.

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