KR20190064721A - Pharmaceutical compositions for preventing or treating menopausal syndrome of women comprising Aristotelia chilensis extract or its enzymatic extract as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating a menopausal symptom, containing an aristoteliachilensis (molina) stuntz extract or an enzymatic hydrolysate thereof as an active ingredient. Particularly, it is confirmed that the aristoteliachilensis (molina) stuntz extract and the enzymatic hydrolysate thereof have an estrogen activity in a cell, and act as an estrogen receptor agonist. Therefore, the aristoteliachilensis (molina) stuntz extract or an enzymatic hydrolysate thereof can be used for an alternative therapy of female hormones for preventing and treating a menopausal symptom derived from estrogen deficiency in a menopausal female.

Description

[0001] The present invention relates to a pharmaceutical composition for preventing or treating menopausal symptoms, which comprises an extract of Mackberry or an enzyme hydrolyzate thereof as an active ingredient. [0002] The present invention relates to a pharmaceutical composition for preventing or treating menopausal symptoms,

The invention lacquer berry (Aristotelia chilensis (Molina) Stuntz) or an enzymatic hydrolyzate thereof as an active ingredient, and a health functional food for preventing or ameliorating menopausal symptoms.

In the life of modern industrialized by the development of science and civilization, the life span of human being is gradually extended, and at the same time, the immune system is degraded due to various stresses, pollution and spontaneous aging phenomenon, and aging progresses. Especially, women suffer physical and mental difficulties due to the menopausal symptoms gradually starting after their forties.

Menopause is a type of endocrine syndrome that refers to a transition period in which physiological function and sexual function are diminished or lost due to a decrease in estrogen, a female hormone resulting from the general and gradual aging of ovarian function. Symptoms of these menopausal symptoms include symptoms due to vascular changes such as facial flushing, tachycardia, sweating or headache, symptoms due to musculoskeletal changes such as myalgia, arthralgia and back pain, symptoms due to genitourinary changes such as urinary frequency or urinary incontinence And depression, depression, loss of concentration, and dizziness. In addition, symptoms such as decreased visual acuity and changes in skin and hair occur, and osteoporosis and cardiovascular diseases caused by hormonal changes Which can lead to health-related diseases.

Therefore, in order to improve the physical and mental health and quality of life of middle-aged women, it was required to develop a therapeutic agent capable of improving the symptoms of menopausal symptoms. To improve these symptoms, hormone replacement therapy and non-steroidal drugs were developed . However, most of these drugs are known to have side effects such as headache and weight gain. Especially, in the case of estrogen replacement therapy, since the hormone is artificially administered to the body, It is known that the risk of seizures, breast cancer and uterine cancer may increase.

Therefore, due to these problems, there is a growing interest in replacing estrogen therapy with natural methods of ingestion in the form of food or additives, and it is necessary to develop a new therapeutic agent for menopausal symptoms that is effective in relieving symptoms of menopause without side effects .

On the other hand, estrogen has a variety of biological functions including increased cell number and differentiation. The biological actions of estrogen exert its activity through two estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ), which belong to the nuclear receptor superfamily. The mechanism of action of the estrogen receptor is the linkage of estrogen and intracellular estrogen receptors to form dimers of receptors and they cause specific estrogen response elements (EREs) located in the promoters of the target gene ) To control the expression of the target gene. Because the two estrogen receptors have different tissue distribution patterns, ligand binding capacity, this difference enables 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 the development of hormone replacement therapy for postmenopausal women and the development of chemotherapy drugs for gynecological cancer. Therefore, if a natural product that activates estrogen receptors is extracted from natural products that do not have side effects on the human body, researches are steadily being carried out because they can be useful for the treatment and prevention of female menopausal symptoms.

The scientific name of maci berry is Aristotelia chilensis (Molina) Stuntz and belongs to Elaeocarpaceae. It is known that the fruit that grows wild in South America such as Chile and has a deep purple color has excellent antioxidative activity including anthocyanin. It is also known that delphinidin, an anthocyanin contained in macchi berry, has an anti-inflammatory effect. However, the effects on women's menopause are not yet known.

Accordingly, the inventors of the present invention have made efforts to develop a therapeutic agent for female menopausal symptoms derived from natural products safe from the human body. As a result, it has been confirmed that the extract of Makibieri and its hydrolyzate have estrogenic activity in cells and act as an estrogen receptor agonist The present invention has been completed based on the finding that the extract of the present invention or the enzyme hydrolyzate thereof can be used in female hormone replacement therapy for the prevention and treatment of menopausal symptoms derived from estrogen deficiency in menopausal women.

Korean Patent No. 1,625,882

Swaran L. Nand, Michael A. Webster, Rodney Baber, Vivienne O'Connor (1998) Bleeding Pattern and Endometrial Changes During Continuous Combined Hormone Replacement Therapy. Obstetrics & Gynecology 91 (5): 678-684. Stefan Nilsson, Sari Makela, Eckardt Treuter, Michel Tujague, Jane Thomsen, Goran Andersson, Eva Enmark, Katarina Pettersson, Margaret Warner, Jan-Ake Gustafsson (2001) Mechanisms of Estrogen Action. Physiological Reviews 81 (4): 1535-1565. Kuiper GG, Carlsson B, Grandien K, Enmark E, Haggblad J, Nilsson S, Gustafsson JA (1997) Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors alpha and beta. Endocrinology 138 (3): 863-870.

An object of the present invention lacquer berry extract (Aristotelia chilensis (Molina) Stuntz) or an enzymatic hydrolyzate thereof as an active ingredient, to provide a pharmaceutical composition for preventing or treating female menopausal symptoms.

It is another object of the present invention to provide a health functional food for preventing or ameliorating female menopausal symptoms comprising an extract of MakiBerry or an enzyme hydrolyzate thereof as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating female menopausal symptoms, which comprises an extract of Machiavelli ( Aristotelia chilensis (Molina) Stuntz) or an enzyme hydrolyzate thereof as an active ingredient.

The present invention also provides a health functional food for preventing or ameliorating female menopausal symptoms comprising an extract of Machiavelli or an enzyme hydrolyzate thereof as an active ingredient.

The inventive Aristotelia chilensis (Molina) Stuntz) extract and its enzymatic hydrolysates have estrogenic activity in cells and act as estrogen receptor agonists, so they are hormone replacement therapy that can improve various symptoms of estrogen deficiency in menopausal women Can be usefully used.

Figure 1 is the estrogen receptor-positive breast cancer cell line MCF-7 estrogen receptor (estrogen receptor; ER) agonists (agonist), ER agonists and ER antagonist (antagonist), or berry lacquer of the present invention (Aristotelia chilensis (Molina) Stuntz) and then confirmed ER-mediated transcriptional activity:
Ethanol: ethanol treatment;
E2 (1 nM): treatment with ER agonist E2 (17β-estradiol) 1 nM;
E2 + ICI: E2 1 nM and ER antagonist ICI (ICI 182,780) 1 μM treatment;
Maqui_100: treatment with 100 [mu] g / ml of the Machiovari extract of the present invention;
Maqui_500: treatment with 500 [mu] g / ml of the maki berry extract of the present invention; And
Maqui_1000: treated with 1000 [mu] g / ml of the extract of the present invention.
Figure 2 shows ER-mediated transcriptional activity after treatment of the ER, ER and ER antagonists, the Makibieri extract of the present invention or an enzymatic hydrolyzate thereof in the estrogen receptor-positive breast cancer cell line MCF-7:
Ethanol: ethanol treatment;
E2 (1 nM): treatment with ER agonist E2 (17β-estradiol) 1 nM;
E2 + ICI: E2 1 nM and ER antagonist ICI (ICI 182,780) 1 μM treatment;
Maqui_100: treatment with 100 [mu] g / ml of the Machiovari extract of the present invention;
Maqui_500: treatment with 500 [mu] g / ml of the maki berry extract of the present invention;
Maqui_1000: treatment with 1000 [mu] g / ml of the barberry extract of the present invention;
Maqui_HE_100: treatment with 100 μg / ml of the enzyme hydrolyzate of the Machiavaryi extract of the present invention;
Maqui_HE_250: Treatment of the enzymatic hydrolyzate of the maki berry extract of the present invention at 250 占 퐂 / ml; And
Maqui_HE_500: Treatment of the enzyme hydrolyzate of the extract of Machiavelli according to the present invention at 500 μg / ml.

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

The present invention relates to a method for the treatment and prevention of arthritis chilensis (Molina) Stuntz) or an enzymatic hydrolyzate thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating female menopausal symptoms.

The active ingredient of the present invention, the makiberry extract, is preferably, but not limited to, prepared by a method comprising the steps of:

1) extracting an extractive solvent in the makibieri; And

2) filtering the extract of step 1).

In the production method of the present invention, the step b) of the step 1) may be cultivated or commercially available.

In the production method of the present invention, it is preferable to use water, an alcohol or a mixture thereof as the extraction solvent in the step 1). As the alcohol, C ₁ to C ₄ lower alcohols are preferably used, and as the lower alcohol, ethanol or methanol is preferably used. The extraction method is preferably, but not limited to, ultrasonic extraction, shaking extraction, Soxhelt extraction or reflux extraction. It is preferable to wash the extraction solvent by 1 to 15 times the amount of the well dried mackerel, and more preferably 2 to 10 times the extraction. The extraction temperature is preferably 20 to 110 DEG C, but is not limited thereto. The extraction time is preferably 10 minutes to 72 hours, more preferably 20 minutes to 48 hours, but is not limited thereto. The number of extraction times is preferably 1 to 5, but is not limited thereto.

In the manufacturing method of the present invention, after the step 2), the method may further include the following steps:

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

4) drying the concentrate of step 3).

In the production method of the present invention, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

In addition, the hydrolyzate of the makibieri extract as an active ingredient of the present invention is preferably, but not limited to, prepared by a method comprising the steps of:

(a) enzymatic hydrolysis of an extract of Machiavelli prepared through steps 1) and 2);

(b) heating the enzyme hydrolyzate of the Makiberry extract of step (a) to inactivate the enzyme; And

(c) adding a solvent to the enzyme hydrolyzate of the makibieri extract of step (b) and filtering.

In the production method of the present invention, the enzyme in step (a) preferably includes at least one of cellulase, hemicellulase, pectinase, and enzymes thereof, It is more preferable to include a thiazine, but it is not limited thereto. In addition, the enzyme may be treated at 0.1 to 15 parts by weight with respect to the weight of the maki berry extract, the enzyme hydrolysis temperature may be 40 to 60 ° C, and the enzyme hydrolysis time may be 1 to 4 hours, but is not limited thereto .

In the production method of the present invention, the enzyme inactivation temperature in step (b) may be 90 to 110 ° C, but is not limited thereto.

In the preparation method of the present invention, in step (c), the solvent may be added to the enzyme hydrolyzate of the Makibieri extract in a ratio of 1: 1 to 3, and the solvent may be water, ethanol or methanol, I never do that.

In addition, after step c), the method may further comprise the steps of:

(d) concentrating the filtrate of step (c) under reduced pressure; And

(e) drying the concentrate of step (d).

The female menopausal symptoms are collectively referred to as symptoms and diseases manifesting in women through menopause as the secretion of estrogen is reduced due to the aging of the ovary. These symptoms include facial flushing, sweating, dry skin, vaginal dryness, vaginal atrophy, , Vaginitis, cystitis, dysuria, rheumatism, intoxication, short-term memory impairment, anxiety, nervousness, memory loss, myalgia, arthralgia or osteoporosis.

The composition has an estrogenic activity and exhibits an estrogen receptor (ER) agonist activity, and has a therapeutic effect on women's menopausal symptoms.

In a specific example of the present invention, the inventors of the present invention measured the ER-mediated transcriptional activity by treating the estrogen receptor-positive breast cancer cell line MCF-7 after preparing the extract of Makibieri and its enzyme hydrolyzate, The ER-mediated transcriptional activity was increased in a concentration-dependent manner in the cells treated with the extract and the enzyme hydrolyzate thereof (see FIGS. 1 and 2).

Therefore, the extract of MakiBerry and the enzyme hydrolyzate thereof of the present invention have an estrogenic activity and exhibit activity as an ER agonist. Therefore, it is an effective ingredient of a pharmaceutical composition for preventing or treating menopausal symptoms derived from estrogen deficiency in menopausal women It can be useful.

The pharmaceutical compositions according to the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the production of pharmaceutical compositions.

The pharmaceutical composition according to the present invention can be administered orally or parenterally, and can be administered orally or parenterally when an external or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, But is not limited thereto.

The pharmaceutical composition according to the present invention can be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like oral preparation, external preparation, suppository and sterilized injection solution according to a conventional method Can be used. Examples of carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added.

A preferable 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 of the patient, sex, and the degree of obesity, but can be appropriately selected by those skilled in the art. For the desired effect, however, in the case of an oral administration agent, it is generally preferred that the composition of the present invention is administered to an adult at a dose of 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg, per 1 kg of body weight per day good. 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 pharmaceutical composition according to the present invention may contain at least one active ingredient which exhibits the same or similar function in addition to the makiberry extract or the enzyme hydrolyzate thereof.

The present invention also provides a health functional food for preventing or ameliorating female menopausal symptoms comprising an extract of Machiavelli or an enzyme hydrolyzate thereof as an active ingredient.

In the present invention, the extract is preferably water, alcohol or a mixture thereof. As the alcohol, C ₁ to C ₄ lower alcohols are preferably used, and as the lower alcohol, ethanol or methanol is preferably used, but it is not limited thereto.

In the present invention, it is preferable that the enzyme hydrolyzate is prepared by treating cellulase, hemicellulase, pectinase and at least one of these enzymes with a makibieri extract, more preferably by treating cellulase and pectinase But is not limited thereto.

In the present invention, the female menopausal symptoms are collectively referred to as symptoms and diseases that appear in women through menopause as the secretion of estrogen is reduced due to aging of the ovary. The symptoms include facial flushing, sweating, skin dryness, vaginal dryness, vaginal atrophy But are not limited to, lower urinary tract atrophy, vaginitis, cystitis, dysuria, rheumatism, intoxication, short-term memory impairment, anxiety, nervousness, memory loss, muscle pain, arthralgia or osteoporosis.

In the present invention, the health functional food has estrogenic activity, exhibits estrogen receptor agonist activity, and has an effect of improving female menopausal symptoms.

The extract of Makibieri of the present invention and the enzyme hydrolyzate thereof have an estrogenic activity and exhibit activity as an ER agonist. Therefore, it is useful as an active ingredient of a health functional food for preventing or ameliorating menopausal symptoms derived from estrogen deficiency in menopausal women Can be used.

The extract of Makibieri or the enzyme hydrolyzate thereof of the present invention can be provided as a food composition by mixing with a pharmaceutically acceptable carrier.

When the maki berry extract or the enzyme hydrolyzate thereof of the present invention is used as a food or beverage additive, the above-mentioned maki berry extract or an enzyme hydrolyzate thereof may be directly added, used together with other foods or food ingredients, and used appropriately according to a conventional method . The mixing amount of the above-mentioned Makibieri extract or the enzyme hydrolyzate thereof can be suitably determined according to its use purpose (prevention, health or therapeutic treatment). In the case of long-term ingestion intended for health and hygiene purposes or for the purpose of controlling health, the above-mentioned Makibieri extract or its enzyme hydrolyzate has no problem in terms of safety, so it can be taken for a long time. There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, drinks, tea, , Alcoholic beverages and vitamin complexes. In case of formulation with beverage, the liquid ingredient to be added in addition to the extract of Makiberry is not limited to this, but it may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of such natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose and polysaccharides such as those commonly used in dextrin and cyclodextrins, , Sorbitol, and erythritol. The ratio 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. Natural flavors such as taumarin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (e.g., saccharin, aspartame, etc.) can be used as flavors other than those described above .

In another aspect, the food composition of the present invention may contain various nutrients, flavors such as vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, , Protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. The food composition of the present invention may also contain pulp for the production of fruit and vegetable drinks. These components may be used singly or in combination, and the proportion of such additives is generally selected in the range of 0.001 to 50 parts by weight based on the total weight of the composition.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation 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 Production Examples.

< Example  1> Maki Berry ( Aristotelia chilensis ( Molina ) Stuntz ) Preparation of extract

Aristotelia chilensis (Molina) Stuntz) The freeze-dried powder was purchased and purchased from Terrasoul Superfoods Inc. of Chile as origin of Chile. Twenty milliliters of water was added to 4 g of the lyophilized Maquisberry dried powder, followed by ultrasonic extraction for 20 minutes. Ultrasonically extracted Mackberry extract was centrifuged at 3,000 rpm for 10 minutes, and the supernatant was separated and used.

< Example  2> Maki Berry ( Aristotelia chilensis ( Molina ) Stuntz ) Enzyme hydrolyzate of extract of  Produce

The following procedure was carried out to prepare an enzyme hydrolyzate of a water extract of macchi berries.

Specifically, a mixture of 100 mg / ml of cellulase and 100 mg / ml of pectinase in a volume ratio of 1: 4 was mixed with 400 mg of the extract of Makibieri water prepared in Example 1 / Ml were mixed at a volume ratio of 1: 1 and stirred in a constant temperature water bath at 50 캜 for 2 hours. After completion of the reaction, the enzyme was inactivated by heating at 100 DEG C and then cooled to room temperature. Then, 100% ethanol was added to the reaction mixture in a volume ratio of 1: 1 (ethanol: mixture), stirred for 10 minutes, and centrifuged at 15,000 rpm for 15 minutes to obtain enzyme hydrolyzate .

< Experimental Example  1> Maki Berry  The estrogen receptor (ER) agonist of the extract agonist ) Active confirmation

The biological action of estrogen is mediated by the binding of estrogen as a ligand to an estrogen receptor (ER), which acts as a transcription factor and binds to an ERE (estrogen response element), thus causing an ERE reporter, It is known that the efficacy of therapeutic agents can be monitored by confirming ER-mediated transcriptional activity using lase. Therefore, in order to investigate whether the extract of Mackerberry acts as an ER agonist and can improve the menopausal symptoms derived from estrogen deficiency in menopausal women, the cells transfected with plasmids containing ERE-luciferase were treated with Mackberry extract ER-mediated transcriptional activity was measured.

Specifically, the estrogen receptor-positive breast cancer cell line MCF-7 purchased from the American Type Culture Collection (ATCC) is supplemented with RPMI 1680 medium supplemented with non-essential amino acids, antibiotics / antimycotics, sodium pyruvate (sodium pyruvate) and 10% fetal bovine serum as a basic medium. And subcultured once a week on average. Cell seeding was carried out 48-60 hours (2-3 days), and the whole serum was treated with dextran-charcoal three times to obtain a stripped serum containing phenol-red Cells were cultured in RPMI 1680 medium. This medium was continuously used until the end of the luciferase assay. The reason for using the medium is to exclude the effect of estrogen receptor interactions due to the estrogenic properties of phenol-red itself and the hormones and growth factors contained in the 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 ERE-luciferase plasmid DNA (0.25 μg / well) Transfection reagent (Invitrogen) was used to transfect the breast cancer cell line.

E2 (17β-estradiol) (ER agonist, 1 nM, Sigma) and ICI 182,780 (Pure ER antagonist, 1 μM, Tocris) were then used as positive controls. E2 and ICI were dissolved in ethanol. In addition, the extract of Makibieri prepared in Example 1 was tested at three concentrations of 100 μg / ml, 500 μg / ml and 1000 μg / ml. Independent experiments confirmed that the final amount of ethanol did not exceed 0.2% of the volume of the medium and that the amount of ethanol had no effect on the final experimental results. On the other hand, the extracts of MakiBerry and the control group were treated with cells and incubated for a further 18-24 hours on average. Then, a luciferase assay was performed. The luciferase activity was evaluated after removing the medium, After washing twice with cold PBS, lysate was extracted with 1 × passive lysis buffer (Promega), and 50 μl of cell-soluble material was added to 50 μl luciferase assay reagent (luciferin, substrate for luciferin and enzyme reaction after mixing the solution, Promega) containing an appropriate buffer, the luminescence intensity that occurred was measured using a SpectraMax M5 ^e (Molecular Device).

As a result, as shown in FIG. 1, it was shown that the cell population treated with the extract of Makibieri showed an increase in ER-mediated transcription activity in a concentration-dependent manner. Therefore, the extract of the present invention had estrogen activity in a concentration- It was confirmed that it exhibited activity as an agonist.

< Experimental Example  2> Maki Berry  Enzyme of extract Hydrolyzate  Identification of estrogen receptor agonist activity

To investigate whether the enzymatic hydrolysates of Makibieri extract could serve as ER agonists to improve the menopausal symptoms induced by estrogen deficiency in menopausal women, the cells transfected with plasmids containing ERE- The enzymatic hydrolysates of the makibieri extracts were treated and the degree of ER-mediated transcription activity was measured.

Example 1 was repeated except that the enzyme hydrolyzate of the extract of Makiberry prepared in Example 2 was treated with 100 μg / ml, 250 μg / ml and 500 μg / ml, instead of the extract of Makiberry. And the degree of ER-mediated transcriptional activity was measured.

As a result, as shown in Fig. 2, the degree of ER-mediated transcriptional activity was also increased in a concentration-dependent manner even in the case of the cell line treated with the enzyme hydrolyzate of the extract of Machiavelli. Therefore, the enzyme hydrolyzate Showed an estrogenic activity and an ER agonist activity in a concentration dependent manner.

Therefore, it was confirmed from the results of <Experimental Example 1> and <Experimental Example 2> that the extract of the present invention and the enzyme hydrolyzate of the present invention have estrogenic activity and function as an ER agonist. Therefore, Enzymatic hydrolysates can be used in female hormone replacement therapies to prevent and treat menopausal symptoms resulting from estrogen deficiency in menopausal women.

Hereinafter, production examples of each preparation according to the present invention will be illustrated. The following Preparation Examples are intended to aid the understanding of the practice of the present invention and do not imply that the process for preparing a formulation according to the present invention is limited to the following Preparation Examples.

&Lt; Preparation Example 1 > Preparation of pharmaceutical preparations

<1-1> Preparation of powder

2 g of the Machiavellian extract of the present invention or the enzyme hydrolyzate thereof

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

<1-2> Preparation of tablets

100 mg of the extract of the present invention or the enzyme hydrolyzate thereof

Microcrystalline cellulose 100 mg

Lactose 100 mg

Starch sodium glyconate 18 mg

Magnesium stearate 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

&Lt; 1-3 > Preparation of capsules

100 mg of the extract of the present invention or the enzyme hydrolyzate thereof

Corn starch 100 mg

100 mg of milk

Magnesium stearate 2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

&Lt; 1-4 >

The extract of the present invention or the enzyme hydrolyzate thereof 1 g

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

<1-5> Preparation of granules

150 mg of the extract of the present invention or its enzyme hydrolyzate

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 캜 to form granules, which were then filled in a capsule.

< Manufacturing example  2> Manufacturing of food

<2-1> Manufacture of confectionery and slices

0.5 to 5.0 parts by weight of the extract of the present invention or its enzyme hydrolyzate of the present invention was added to wheat flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture to prepare a food for health promotion.

<2-2> Manufacture of dairy products

5 to 10 parts by weight of the extract of the present invention or the enzyme hydrolyzate thereof 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> Solar  Produce

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh. Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The Makiberry extract of the present invention or its enzyme hydrolyzate was concentrated under reduced pressure in a vacuum concentrator, dried by spraying, and dried with a hot air drier, and pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder. The grains, seeds and SBE dry powders prepared above were blended in the following proportions.

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

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

(3 parts by weight) of the extract of Makibieri of the present invention or the enzyme hydrolyzate thereof,

(0.5 part by weight), and rhizome (0.5 part by weight)

< Manufacturing example  3> Manufacturing of beverage

The extract of Makibieri of the present invention or its enzyme hydrolyzate 1000 mg

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water was added to a total of 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 ° C for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, It is used in the manufacture of health food of the invention.

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Claims (13)

Aristotelia chilensis (Molina) Stuntz) or an enzymatic hydrolyzate thereof as an active ingredient for the prevention or treatment of female menopausal symptoms.
The method according to claim 1, wherein the Makiberry extract is water, C ₁ To C ₄ lower alcohols or a mixed solvent thereof.
[3] The pharmaceutical composition according to claim 2, wherein the extract is extracted with at least one solvent selected from the group consisting of water, methanol, ethanol, and a mixed solvent thereof.
[Claim 2] The method according to claim 1, wherein the enzyme hydrolyzate is prepared by treating at least one enzyme selected from the group consisting of Cellulase, Hemicellulase, and Pectinase, A pharmaceutical composition for the prevention or treatment of female menopausal symptoms.
The method of claim 1, wherein the female menopausal symptoms are selected from the group consisting of facial flushing, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urinary tract atrophy, vaginitis, cystitis, dysuria, Decline, muscle pain, arthralgia, and osteoporosis. The pharmaceutical composition for preventing or treating female menopausal symptom.
The pharmaceutical composition according to claim 1, wherein the composition is an estrogen receptor agonist.
The pharmaceutical composition according to claim 1, wherein the composition has an estrogenic activity.
A health functional food for preventing or ameliorating female menopausal symptoms, comprising an extract of Makiberry or an enzyme hydrolyzate thereof as an active ingredient.
9. The method of claim 8, wherein the female menopausal symptoms are selected from the group consisting of facial flushing, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urinary tract atrophy, vaginitis, cystitis, dysuria, Depression, muscular pain, arthralgia, and osteoporosis. The health functional food for preventing or ameliorating female menopausal symptoms.
A food composition for preventing or ameliorating a female menopausal symptom, which comprises an extract of Mackberry or an enzyme hydrolyzate thereof as an active ingredient.
11. The method of claim 10, wherein the female menopausal symptoms are selected from the group consisting of facial flushing, sweating, dry skin, vaginal dryness, vaginal atrophy, lower urinary tract atrophy, vaginitis, cystitis, dysuria, Decongestion, myalgia, joint pain, and osteoporosis. The composition for preventing or improving female menopausal symptom.
1) extracting an extractive solvent in the makibieri; And
2) filtering the extract of step 1).
a) treating enzyme-treated enzyme-hydrolyzed Mackberry extract prepared by the method of claim 12;
b) heating the enzyme hydrolyzate of the maki berry extract of step a) to inactivate the enzyme; And
and c) adding an enzyme hydrolyzate of the maki berry extract of step b) to a solvent, followed by filtration, to obtain an enzyme hydrolyzate of a makibieri extract.

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