KR20210089370A - Composition for Preventing Hair Loss or Improving Hair Growth Containing Extract of Fermented Cheju Pureunkong - Google Patents

Abstract

The present invention relates to a composition for preventing hair loss or improving hair growth containing an extract of fermented Jeju green beans and, more specifically, to a composition having an effect of preventing hair loss or improving hair growth by containing an extract of Jeju green beans fermented with Bacillus sp. strains.

Description

Composition for Preventing Hair Loss or Improving Hair Growth Containing Extract of Fermented Cheju Pureunkong, containing an extract of fermented Jeju green beans

The present invention relates to a composition for preventing hair loss or improving hair growth containing an extract of fermented Jeju green beans, and more specifically, to a composition having an effect of preventing hair loss or improving hair growth by containing an extract of Jeju green beans fermented with a Bacillus sp. it's about

The scalp is composed of epidermis, dermis and subcutaneous tissue, and the epidermis, the outermost tissue, functions to protect sensitive tissues of the body from damage. However, due to environmental factors affecting the scalp, for example, stress and lack of influence due to external stimuli such as temperature change, humidity change, exposure to pollutants, etc., scalp function may be lost, and hair loss may also be affected. .

Tissues associated with hair loss include sebaceous glands, keratinocytes of the epidermis and hair follicles, dermal papilla cells, sweat glands, and root sheath of the scalp hair. follicle), etc.

In male expression, it is known that testosterone is involved in skeletal muscle increase, spermatogenesis, etc., and dihydrotestosterone (DHT) is involved in the tissue in acne, sebum increase, hair loss, and prostatic hyperplasia, etc. ( Thiboutot et al., J. Invest. Dermatol ., 105(2):209-14, 1995).

Androgenetic alopecia is a phenomenon that is caused by a male hormone called testosterone. When this testosterone is converted into dihydrotestosterone, a stronger hormone by an enzyme called 5-alpha reductase (5-alpha-reductase), this hormone Acting on this hair follicle, it induces the hair follicle from the growth phase to the degeneration phase, causing hair loss. Therefore, a method of inhibiting the production of DHT by 5 alpha-reductase is mainly used to treat androgenetic alopecia.

On the other hand, it has been found that finasteride, a 5-alpha reductase inhibitor, improves hair loss by reducing the hair loss hormone DHT in hair loss patients (Van Neste et al. , 2000), through which 5-alpha reductase is As an important target for the treatment of hair loss, it is used in the development of a treatment for hair loss, and a study to find a pharmacologically active substance to treat alopecia by inhibiting the activity of 5-alpha reductase, a biological enzyme involved in the reaction of testosterone to dihydrotestosterone A lot is being done

Gynecomastia is mainly caused by a decrease in the amount of estrogen after menopause. Minoxidil or estrogen is mainly used as a treatment for female pattern alopecia. Alopecia areata is caused by autoimmune diseases, mental stress, or genetic predisposition. Alopecia areata has a fundamentally different cause from androgenetic alopecia, and the treatment is also different, so a method of treating adrenocortical hormones, applying minoxidil to the affected area, or artificially stimulating the affected area is used.

For these complex causes of hair loss, ingredients for the purpose of promoting blood circulation, suppressing the action of male hormones, and strengthening the function of hair roots are sold as products on the market, but none with a distinct effect has been found yet, and the problem of side effects is also raised For example, in the case of minoxidil, a sticky feeling of use and side effects causing irritation to the skin have been reported, and in the case of finasteride, which is currently used as a formulation for oral administration, side effects such as sexual dysfunction have been reported according to its ingestion, The effect could be expected only by oral administration, so there were many inconveniences in use.

For the prevention and treatment of such hair loss, hormone replacement therapy is used as the most effective method. However, when hormones such as estrogen are administered for a long time, there is a risk of cancer such as breast cancer and uterine cancer, and there are problems in that side effects such as headache and weight gain occur.

On the other hand, isoflavone of soybean, which is an estrogen-like substance (phytoestrogen) extracted from plants, is commonly used for the purpose of improving hair loss, but issues regarding effectiveness and safety are continuously being raised. For this reason, there is an urgent need for the development of a treatment for hair loss using a safer and more effective natural substance.

Jeju green beans (scientific name: Glycine max , common name: Soybean, cultivar name: Pureunkong) are called 'purin poison peas' in Jeju, which means blue egg-shaped beans. It is also called 'jang-kong' because it is used to make soy sauce. Jeju green beans are a native resource produced only in a part of the southern part of Jeju Island, and are in danger of extinction due to difficulties in preserving seeds. Jeju green beans are taller than regular beans, so they are vulnerable to wind and have a long cultivation period. If planted on land other than Jeju, the quality and ingredients may deteriorate. Jeju green soybeans contain much more isoflavones than ordinary soybeans, and the traditional doenjang made with Jeju green soybeans has many complex bacteria, fungi, and enzymes generated from nature, such as thrombolytic ability, anticancer effect, etc., compared to the improved doenjang sold in the market. It is known to have excellent various effects.

Poly-γ-glutamic acid (γ-PGA) is an anionic amino acid polymer in which the γ-carboxyl group of glutamic acid and the α-amino group of glutamic acid are combined with amide. It is produced from Bacillus genus strains isolated from Cheonggukjang, natto, a traditional fermented soybean food in Japan, and Kinema, a traditional fermented soybean food in Nepal. Polygamma-glutamic acid produced from Bacillus sp. strain is edible, water-soluble, anionic and biodegradable.

7,8,4'-Trihydroxyisoflavone (7,8,4'-Trihydroxyisoflavone, structural formula: C ₁₅ H ₁₀ O ₅ ) is one of the representative isoflavones daidzin and daidzein ) is a major metabolite of 7,8,4'-trihydroxyisoflavones are known to be contained in soybean paste, a fermented soybean food (Park, JS., et al ., Bioorganic & Medicinal Chemistry Letters , 20:1162-1164, 2010; Roh, CH., Biomolecules , 4(4):1093-1101, 2014).

In the case of minoxidil, one of the hair growth promoting drugs approved by the Food and Drug Administration (FDA), it was initially developed as a vasodilator for the treatment of high blood pressure, but as a side effect of hirsutism was reported, it is now used as a hair growth agent. is becoming Although the specific mechanism of action regarding the hair growth effect of minoxidil has not yet been clearly elucidated, K _ATP -channel opening, increased nutrient supply through vasodilation, and activation of the β-catenin pathway in human dermal papilla cells It is thought to induce hair growth. Human dermal papilla cells are mesoderm-derived fibroblasts that cycle through the hair cycle through secretion of proteins such as insulin-like growth factor-1 (IGF-1), hepatocyte growth factor (HGF) and fibroblast growth factor-7 (FGF-7). It is reported to have an effect on the dermal papilla, and minoxidil is known to have an effect of proliferating the growth of human dermal papilla cells by lowering apoptosis of human dermal papilla cells.

Therefore, measuring the 5-alpha reductase activity inhibitory effect of a substance and the growth and proliferation effect of human dermal papilla cells can be an effective indicator for evaluating how effectively the substance can treat hair loss and improve hair growth.

On the other hand, according to the prior art, the white hair, hair loss and dandruff inhibitory efficacy of a composition containing a ginseng extract and a green tea extract is disclosed (refer to Korean Patent Publication No. 10-2015-0030095 [2015.03.19.]), Jeju green beans Although the ceramide biosynthesis promoting effect of the composition containing the extract of has been disclosed (refer to Korean Patent Publication No. 10-2019-0017084 [2019.02.20.]), the extract of fermented Jeju green beans or fermented with a Bacillus sp. The contents of the composition for promoting hair growth and improving symptoms of hair loss, containing the extract of Jeju green beans as an active ingredient, are not disclosed.

Under this technical background, the present inventors have developed a composition that exhibits the effect of improving hair growth and hair loss symptoms by containing as an active ingredient an extract of fermented Jeju green beans, preferably an extract of Jeju green beans fermented with Bacillus sp. .

Accordingly, the present inventors have made diligent efforts to develop a composition containing a plant-derived extract effective for preventing hair loss or improving hair growth, as a result of which the extract of fermented Jeju green beans, preferably the extract of Jeju green beans fermented with Bacillus sp. By promoting the expression of VEGF (Vascular Epidermal Growth Factor) and HGF (Hepatocyte Growth Factor), which are growth factors of

Accordingly, the main object of the present invention is to provide a composition containing, as an active ingredient, an extract of fermented Jeju green beans having excellent hair growth promotion and hair loss prevention effects, preferably an extract of Jeju green beans fermented with a Bacillus sp.

In order to achieve the above object, the present invention provides a composition for preventing hair loss or improving hair growth containing an extract of fermented Jeju green beans, preferably an extract of Jeju green beans fermented with Bacillus sp. It may be characterized as a pharmaceutical composition for preventing hair loss or improving hair growth, a health functional food composition, and a cosmetic composition.

The present invention also provides a method for producing a fermented Jeju green bean extract,

(a) inoculating Bacillus sp. strain into Jeju green beans and fermenting them to obtain a fermented product of Jeju green beans;

(b) preparing a reactant by adding a solvent to the fermented product and then drying the filtrate of the reaction product obtained by incubation to obtain dry matter 1 by filtration;

(c) drying the filtrate of the reaction product obtained by adding a solvent to Jeju green beans to prepare a reaction product and leaving it to stand to obtain dry matter 2 (dry matter 2); and

(d) mixing (combined use) the filtered dried product 1 and the filtered dried product 2 to obtain a fermented Jeju green bean extract;

It provides a method for producing a fermented Jeju green bean extract, characterized in that the content of each of the Jeju green beans of step (a) and the Jeju green beans of step (c) is the same.

When the composition containing the extract of fermented Jeju green beans according to the present invention is used, the effect of preventing hair loss or improving hair growth by promoting the expression of VEGF and HGF, which are signals for promoting hair growth, in tissues associated with hair loss including dermal papilla cells have.

1 is a graph showing the degree of proliferation of human dermal papilla cells treated with an extract of Jeju green beans fermented with a Bacillus sp. strain according to the present invention.
Figure 2 is a graph showing the expression level of VEGF in human dermal papilla cells treated with an extract of Jeju green beans fermented with a Bacillus sp. strain according to the present invention.
3 is a graph showing the expression level of HGF in human dermal papilla cells treated with an extract of Jeju green beans fermented with a Bacillus sp. strain according to the present invention.
Figure 4 is a graph showing the hair growth length of the human hair follicle organ treated with the extract of Jeju green beans fermented with the Bacillus sp. strain according to the present invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is those well known and commonly used in the art.

As used herein, the term “about” used to express length, area, volume, time (period), concentration, content, temperature, etc. means that there is a tolerance of up to 10% in the corresponding numerical value or numerical range.

The extract of fermented Jeju green beans according to the present invention, preferably the extract of Jeju green beans fermented with a Bacillus sp. strain, is a substance that promotes hair growth by regulating the expression of VEGF and HGF, and promotes hair growth through the regulation of male hormones Since it is not involved in male hormones, severe hair loss and female pattern hair loss that have not been resolved by methods of suppressing male hormones can be improved.

Accordingly, in one aspect, the present invention provides a pharmaceutical composition for preventing hair loss or improving hair growth, a health functional food composition or a cosmetic comprising, as an active ingredient, an extract of fermented Jeju green beans, preferably an extract of Jeju green beans fermented with Bacillus sp. to the composition.

In another aspect, the present invention relates to a method for producing a fermented Jeju green bean extract.

Specifically,

(a) inoculating Bacillus sp. strain into Jeju green beans and fermenting them to obtain a fermented product of Jeju green beans;

(b) preparing a reactant by adding a solvent to the fermented product and then drying the filtrate of the reaction product obtained by incubation to obtain dry matter 1 by filtration;

(c) drying the filtrate of the reaction product obtained by adding a solvent to Jeju green beans to prepare a reaction product and leaving it to stand to obtain dry matter 2 (dry matter 2); and

(d) mixing (combined use) the filtered dried product 1 and the filtered dried product 2 to obtain a fermented Jeju green bean extract;

It relates to a method for producing a fermented Jeju green bean extract, characterized in that the content of each of the Jeju green beans of step (a) and the Jeju green beans of step (c) is the same.

The content of the Bacillus sp. strain inoculated into Jeju green beans in step (a) is 0.1 x 10 ⁸ CFU (colony forming unit) / mL ~ 10.0 x 10 ⁸ CFU / mL, preferably 1.0 x 10 ⁸ CFU / mL ~ 2.0 x 10 ⁸ CFU/mL. When the content of the Bacillus sp. strain is ^{less than 0.1 x 10 8} CFU/mL, the fermentation efficacy may be low, and ^{even if the content of the Bacillus sp. strain exceeds 10.0 x 10 8} CFU/mL, the fermentation effect may not further increase.

Fermentation in step (a) may be performed at a temperature of 35.0 to 38.0 ° C., preferably 36.5 to 37.5 ° C. for 24 to 96 hours, preferably 69 to 75 hours, more preferably 71 to 73 hours. When the fermentation temperature is less than 35.0°C, the fermentation efficacy may be low, and even if the fermentation temperature exceeds 38.0°C, the fermentation efficacy may not further increase.

The weight ratio of the fermented product and the solvent constituting the reactant in step (b) may be 1:1 to 9, preferably 1:1 to 4, more preferably 1:1 to 2, and if it is out of the above weight ratio The yield of the reaction product may decrease.

Obtaining the reaction product in step (b) is made by leaving the reactant composed of the fermented product and the solvent at a temperature of 85.0 to 95.0° C., preferably 88.0 to 92.0° C. for 2 to 4 hours, preferably 2 to 3 hours. can When the temperature at the time of obtaining the reaction product is less than 85.0°C, the yield of the reaction product may be low, and even if the temperature at the time of obtaining the reaction product exceeds 95.0°C, the yield of the reaction product may not increase further.

The weight ratio of Jeju green beans and solvent constituting the reactant in step (c) may be 1:1 to 9, preferably 1:1 to 4, more preferably 1:1 to 2, and if it is out of the above weight ratio The yield of the reaction product may decrease.

To obtain the reaction product in step (c), the reaction product composed of Jeju green beans and a solvent is heated at a temperature of 20.0 to 25.0° C., more preferably 21.0 to 23.0° C. for 24 to 96 hours, preferably 69 to 75 hours, more preferably This can be done by leaving it to stand for 71 to 73 hours. When the temperature at the time of obtaining the reaction product is less than 20.0°C, the yield of the reaction product may be low, and even if the temperature at the time of obtaining the reaction product exceeds 25.0°C, the yield of the reaction product may not increase further.

The Jeju green soybean extract fermented in step (d) is 1 to 9: 9 to 1, preferably 1 to 4: 4 to 1, more preferably 1 to 2, based on the weight of the filtered dried product 1 and the filtered dried product 2 : It can be characterized as obtained by mixing (combined use) in a weight ratio of 2-1. If the weight ratio is out of the above weight ratio, the synergistic effect of growth factors (VEGF and/or HGF) and cell proliferation increasing effect of the dermal papilla cells by the fermented Jeju green bean extract may be low, and the hair growth effect of the human hair follicle organ may also be reduced. .

The filtration in steps (b) and (c) may be performed through a 0.1 to 0.75 μm filter, preferably a 0.2 to 0.5 μm filter, more preferably a 0.3 μm filter, but is not limited thereto.

In the present invention, the Bacillus sp. strain is Bacillus subtilis ( Bacillus subtilis ), Bacillus cereus ( Bacillus cereus ), Bacillus natto ( Bacillus natto ), Bacillus licheniformis ( Bacillus licheniformis ), Bacillus pumilus ( Bacillus pumilus ) ) and Bacillus thuringiensis ( Bacillus thuringiensis ) It may be at least one selected from the group consisting of, but is not limited thereto.

In one aspect of the present invention, "Jeju green beans" may be raw Jeju green beans, a pulverized product of raw Jeju green beans, a dried product of Jeju green beans, or a dry pulverized product of Jeju green beans, but is not limited thereto. In addition, the Jeju green beans used in the present specification are not limited in the method of obtaining them, and may be cultivated and used or may be purchased and used commercially, and some or all of the above-ground part or root part of the herb may be used. More specifically, at least one selected from the group consisting of fruits (seeds), leaves, stems, roots, flowers and outposts of the herbaceous Jeju green bean may be used, and more specifically, seeds may be used. In one aspect of the present invention, in the case of Jeju green beans, it is not necessarily manufactured through drying and is not limited as long as it is a raw material in a form suitable for extracting the active ingredients of Jeju green beans.

In one aspect of the present invention, the extract of fermented Jeju green beans is a final product (filtration) derived from a reaction product obtained by using one solvent selected from the group consisting of water, organic solvents, and mixtures thereof. It may be characterized in that it is a mixture of the dried product 1) and the final product (filtered dried product 2) derived from the reaction product obtained by using one solvent selected from the group consisting of water, organic solvents, and mixtures of Jeju green beans. .

In one aspect of the present invention, water includes distilled water or purified water, and the organic solvent is at least one selected from the group consisting of C1-C6 lower alcohols, acetone, ether, ethyl acetate, diethyl ether, ethylmethyl ketone, and chloroform. including, but not limited to. Specifically, the alcohol may be methanol or ethanol.

As used herein, the term "improve" or "alleviate" refers to the reduction or elimination of a condition, disease, disorder or phenotype, including an abnormality or symptom, syndrome. Here, the improvement or alleviation of hair loss means preventing, improving or alleviating hair loss diseases by promoting the expression of VEGF and HGF, which are signals for promoting hair growth, in tissues associated with hair loss, including dermal papilla cells.

As used herein, "treatment" refers to any action in which the symptoms of hair loss disease are improved or cured, and more specifically, the disease or disease to which the term is applied, or one or more symptoms of the disease or disease is reversed. or to alleviate, inhibit its progression, or prevent.

In the present invention, the extract of fermented Jeju green beans, preferably the extract of Jeju green beans fermented with Bacillus sp. strain, is based on the total weight of the composition, 0.1 to 20.0% by weight, preferably 0.5 to 10.0% by weight, more Preferably, it may be characterized in that it is contained in an amount of 1.0 to 5.0% by weight. If the content of the extract of fermented Jeju green beans or the extract of Jeju green beans fermented with Bacillus sp. strain is less than 0.1% by weight, it is difficult to expect the effect of promoting hair growth and preventing hair loss, and if it exceeds 20.0% by weight, the stability of the formulation is lowered and the extract It is expected that the effect will be lowered by affecting the component stability of

In the present invention, the extract of fermented Jeju green beans, preferably the extract of Jeju green beans fermented with Bacillus sp. strain, can promote the expression of VEGF and/or HGF, which are growth factors of dermal papilla cells, more preferably human Promotes the expression of VEGF and/or HGF in human dermal papilla cells derived from the skin tissue of

In the present invention, the extract of fermented Jeju green beans, preferably the extract of Jeju green beans fermented with Bacillus sp. strain, is 1-50 ppm, preferably 10-50 ppm, most preferably 20 ppm (w/w) basis. It may be characterized by promoting the expression of VEGF and HGF in dermal papilla cells at ~50ppm.

In the present invention, the extract of the fermented Jeju green beans, preferably the extract of the Jeju green beans fermented with a Bacillus sp. strain, may be characterized in that the hair of the hair follicle organ grows.

In the present invention, the extract of fermented Jeju green beans, preferably the extract of Jeju green beans fermented with Bacillus sp. strain, promotes the expression of VEGF and/or HGF, which are growth factors of dermal papilla cells, thereby promoting hair growth and/or hair loss symptoms. It may be characterized as having an improvement effect.

In the present invention, the extract of fermented Jeju green beans, preferably the extract of Jeju green beans fermented with Bacillus sp. strain, contains poly-γ-glutamic acid (γ-PGA) and 7,8,4'-tree Hydroxyisoflavone (7,8,4'-Trihydroxyisoflavone, structural formula: C ₁₅ H ₁₀ O ₅ ) It may be characterized in that it contains at least one selected from the group consisting of.

The fermented Jeju green soybean extract contains 1,000 to 4,500 ppm, preferably 2,000 to 3,000 ppm of polygamma-glutamic acid, based on the total ppm (w/w) of the extract, and 7,8,4'-trihydroxy It may be characterized in that it contains isoflavones in an amount of 0.1 to 20 ppm, preferably 1 to 2 ppm.

The pharmaceutical composition containing the extract of the fermented Jeju green beans of the present invention, preferably the extract of the Jeju green beans fermented with a Bacillus sp. strain, contains a preservative, a stabilizer, a wetting agent or emulsification promoter, a salt and/or a buffer for osmotic pressure control It may further contain pharmaceutical adjuvants and other therapeutically useful substances (carriers, excipients, diluents, etc.), and may be formulated in various oral or parenteral administration forms according to conventional methods.

The term “carrier” is defined as a compound that facilitates the addition of the compound into a cell or tissue. For example, dimethylsulfoxide (DMSO) is a commonly used carrier that facilitates the introduction of many organic compounds into cells or tissues of living organisms.

The term "excipient" is a substance added to make the drug easier to eat or to have a certain color and shape when the amount of the main drug is small in the preparation process of tablets or pills, and lactose or starch is mainly used do.

The term "diluent" is defined as a compound that not only stabilizes the biologically active form of the compound of interest, but is diluted in water which will dissolve the compound. Salts dissolved in buffer solutions are used as diluents in the art. A commonly used buffer solution is phosphate buffered saline because it mimics the salt state of human solutions. Because buffer salts can control the pH of a solution at low concentrations, buffer diluents rarely modify the biological activity of a compound.

As used herein, a composition containing an extract of fermented Jeju green beans, preferably an extract of Jeju green beans fermented with a strain of Bacillus sp., is suitable for human patients by themselves or in combination with other active ingredients as in combination therapy or as appropriate. It can be administered as a pharmaceutical composition mixed with a carrier or excipient.

Carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The oral administration agents include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, powders, powders, fine granules, granules, pellets, etc., and these formulations contain surfactants in addition to active ingredients. , diluents (such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (such as silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycol). Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, optionally starch, agar, alginic acid or its sodium salt It may contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavoring agents, and sweetening agents, such as. The tablet may be prepared by a conventional mixing, granulating or coating method.

In addition, the parenteral administration agent may be in the form of injections, drops, ointments, lotions, gels, creams, sprays, suspensions, emulsions, suppositories, patches, etc., but is not limited thereto. it is not

The pharmaceutical composition according to an embodiment of the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, or the like. The pharmaceutical composition according to an embodiment of the present invention may be administered topically to the scalp, for example.

In addition, the pharmaceutically acceptable dose of the active ingredient, that is, the dosage may be determined by the age, sex, and weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathology, the route of administration, and the judgment of the prescriber. It will be different. Dosage determination based on these factors is within the level of one of ordinary skill in the art. The dosage is, for example, from one to several times to be administered in an amount of 0.01 to 5000 mg, preferably 0.1 to 2,000 mg, more preferably 0.5 to 500 mg, and most preferably 1 to 100 mg per 1 kg of body weight per day. It can be administered in divided doses, but the dosage does not limit the scope of the present invention in any way.

The extract of the fermented Jeju green beans of the present invention, preferably the extract of the Jeju green beans fermented with a Bacillus sp. strain, is a natural substance and therefore has no toxicity, so it can be continuously used in large amounts as pharmaceuticals.

In the pharmaceutical composition suitable for use in the present invention, active ingredients, including extracts of fermented Jeju green beans, preferably extracts of Jeju green beans fermented with Bacillus sp. strain, are contained in an amount effective to achieve its intended purpose. compositions are included. More specifically, a therapeutically effective amount means an amount of a compound effective to prolong the survival of the subject being treated or to prevent, alleviate or ameliorate the symptoms of a disease. Determination of a therapeutically effective amount is within the ability of one of ordinary skill in the art, particularly in light of the detailed disclosure provided herein.

The therapeutically effective amount of the fermented Jeju green soybean extract used in the methods of the present invention, preferably the Jeju green soybean extract fermented with a Bacillus sp. strain, and the composition (compounds) containing the same as an active ingredient is determined by cell culture can be initially measured from For example, a dose can be calculated in an animal model to obtain a circulating concentration range that includes an _{IC 50} (half maximal inhibitory concentration) or EC _{50 (half maximal effective concentration) determined in cell culture.} Such information can be used to more accurately determine useful doses in humans.

The toxicity and therapeutic efficacy of the extract of fermented Jeju green beans described herein, preferably the extract of Jeju green beans fermented with a Bacillus sp. strain, or a composition (compounds) containing the same as an active ingredient, is, for example, LD _{To determine 50} (lethal dose for 50% of population), ED ₅₀ (dose having a therapeutic effect on _{50% of population), IC 50} (dose having a therapeutically inhibitory effect on 50% of population), cell culture Alternatively, it can be estimated by table-constraint procedures in laboratory animals. The dose ratio between toxicity and therapeutic effect is the therapeutic index and it can be expressed as the ratio between _{LD 50} and ED ₅₀ (or IC _{50 ).} Compounds exhibiting high therapeutic indices are preferred. Data obtained from these cell culture assays can be used to estimate a range of doses for use in humans. The dosage or application of such compounds is preferably within a range of circulating concentrations that include the _{ED 50} (or IC _{50 ) with little or no toxicity.}

In the present invention, 'functional food' or 'functional food' refers to an extract of fermented Jeju green beans of the present invention, preferably an extract of Jeju green beans fermented with a Bacillus sp. means food.

Functionality can be roughly divided into physical properties and physiological functionality. When the extract of fermented Jeju green beans of the present invention, preferably the extract of Jeju green beans fermented with Bacillus sp. strain, is added to general food, physical properties and physiological functionality of general food This will be improved, and the present invention comprehensively defines the food of such improved function as 'functional food (health functional food)' or 'functional food (health functional food)'.

In the present invention, the term 'health functional food' refers to food manufactured and processed using raw materials or ingredients useful for the human body in accordance with Act No. 10219 of the Health Functional Food Act, and provides nutrients for the structure and function of the human body. It refers to ingestion for the purpose of obtaining useful effects for health purposes, such as controlling or physiological effects.

The health functional food containing the extract of the fermented Jeju green beans of the present invention, preferably the extract of the Jeju green beans fermented with a Bacillus sp. strain, increases the main effect within the range that does not impair the main effect of the present invention. It may contain other ingredients that can give an effect. For example, in order to improve physical properties, additives such as fragrances, pigments, bactericides, antioxidants, preservatives, humectants, thickeners, inorganic salts, emulsifiers, and synthetic polymers may be further included. In addition, auxiliary components such as water-soluble vitamins, oil-soluble vitamins, high molecular peptides, high molecular weight polysaccharides and seaweed extract may be further included. The above ingredients can be appropriately selected and formulated by those skilled in the art without difficulty depending on the formulation or purpose of use, and the amount added may be selected within a range that does not impair the purpose and effect of the present invention. In addition, the composition of the present invention itself can be used in the form of an additive for other foods.

The formulation of the health functional food containing the extract of the fermented Jeju green beans of the present invention, preferably the extract of Jeju green beans fermented with a Bacillus sp. strain is not particularly limited, but, for example, tablets, granules, drinks, beverages, solutions , can be formulated in various forms such as emulsions, viscous mixtures, tablets, powders, and the like. In addition, when administering the health functional food, it may be administered by various methods such as simple drinking, injection administration, spray method, or squeeze method.

According to one embodiment of the present invention, the composition containing the extract of fermented Jeju green beans of the present invention, preferably the extract of Jeju green beans fermented with Bacillus sp. strain, is formulated as a cosmetic formulation to promote hair growth. It can be used in various ways, etc.

The cosmetic composition containing the extract of the fermented Jeju green beans of the present invention, preferably the extract of the Jeju green beans fermented with a Bacillus sp. strain, is not particularly limited in formulation, and may be appropriately selected according to the intended purpose. For example, softening lotion (skin lotion and milk lotion), nourishing lotion, essence, nourishing cream, massage cream, pack, gel, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, Body lotion, body cream, body oil, body essence, hair tonic, hair conditioner, hair essence, hair lotion, hair nourishing lotion, hair shampoo, hair conditioner, hair treatment, hair cream, hair nourishing cream, hair moisturizing cream, hair Massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair drying agent, hair preservative, hair dye, hair wave agent, hair bleach, hair gel, hair glaze, hair dresser , hair lacquer, hair moisturizer, hair mousse and may be prepared in any one or more formulations selected from the group consisting of hair spray, but is not limited thereto, preferably hair tonic, hair conditioner, hair essence, hair lotion, hair Nutritional lotion, hair shampoo, hair conditioner, hair treatment, hair cream, hair nutrition cream, hair moisture cream, hair massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair It may be formulated as a desiccant, hair preservative, hair dye, wave agent for hair, hair bleach, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, hair mousse or hair spray.

When the composition containing the extract of the fermented Jeju green beans of the present invention, preferably the extract of the Jeju green beans fermented with a Bacillus sp. strain, is formulated as a cosmetic formulation, the composition of the present invention may be used in cosmetics in addition to the extract. All of the components may be included, for example, a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, sphingolipids, and seaweed extract.

As the macromolecular peptide, any one can be selected as long as it can be formulated in cosmetics, but preferably collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin and the like can be selected. Polymeric peptides can be purified and obtained by conventional methods such as purification from a microbial culture solution, enzyme method or chemical synthesis method, or can be purified and used from natural products such as dermis of pigs or cattle, silk fibers of silkworms, etc.

As the high molecular weight polysaccharide, any one can be selected as long as it can be formulated in cosmetics, but preferably, hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfuric acid or a salt thereof (sodium salt, etc.) can be selected. For example, chondroitin sulfate or a salt thereof can be purified and used from mammals or fish.

As the sphingolipid, any one can be selected as long as it can be formulated in cosmetics, but preferably ceramide, phytosphingosine, sphingoglycolipid, and the like can be selected. The sphingolipid can be purified by a conventional method from mammals, fish, shellfish, yeast or plants, or can be obtained by chemical synthesis.

In the cosmetic of the present invention, in addition to the above essential ingredients, other ingredients commonly formulated in cosmetics may be blended as needed. At this time, the ingredients include oil and fat ingredients, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohol, pigments, fragrances, blood circulation promoters , a cooling agent, a limiting agent, purified water, etc. may be used.

In addition, the compounding component that may be added is not limited thereto, and any of the above components can be compounded within a range that does not impair the purpose and effect of the present invention, but based on the total weight of the composition, 0.001 to 50% by weight , preferably 0.01 to 5% by weight, more preferably 0.01 to 3% by weight.

Components included in the composition formulated with the cosmetic formulation of the present invention may include components commonly used in cosmetics in addition to the extract as an active ingredient, for example, stabilizers, solubilizers, vitamins, pigments and fragrances and such conventional adjuvants and carriers.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as being limited by these examples.

[Example 1] Preparation of extract derived from Jeju green beans fermented with Bacillus sp.

First, 10 g of Jeju green beans with Bacillus subtilis ( Bacillus subtilis ) 1.0 x 10 ⁸ CFU (colony forming unit) / mL (the strain obtained by culturing in ATCC Medium 3 is suspended in PBS [phosphate buffered saline]) After inoculation, it was fermented at a temperature of about 37° C. for about 72 hours to obtain a fermented product of Jeju green beans.

Purified water is added to the fermented product to prepare a reactant so that the weight ratio of the fermented product and purified water is 1:1, and then incubated at a temperature of about 90° C. for about 3 hours to obtain a reaction product, the reaction product was filtered through a 0.3 μm filter, and the obtained filtrate was dried to obtain a filtered dry matter 1 (dry matter 1).

Next, 10 g of Jeju green beans and a 95% aqueous ethanol solution (ethanol: purified water volume ratio = 95: 5) were prepared so that the weight ratio of the reactant was 1:1, and then left at a temperature of about 22° C. for about 72 hours. The reaction product was filtered through a 0.3 μm filter, and the filtrate obtained was dried to obtain dry matter 2 by filtration.

The filtered dried product 1 and the filtered dried product 2 were mixed at a ratio of 1:1 by weight to prepare a fermented Jeju green soybean-derived extract (fermented Jeju green soybean extract).

[ comparative example One] bacillus fermented with isolates Big head Preparation of Derived Extracts

An extract was prepared in the same manner as in Example 1 using soybeans instead of Jeju green beans.

[ comparative example 2] bacillus fermented with isolates Seoritae Preparation of Derived Extracts

An extract was prepared in the same manner as in Example 1 using Seoritae instead of Jeju green beans.

[ comparative example 3] bacillus fermented with isolates green beans Preparation of Derived Extracts

An extract was prepared in the same manner as in Example 1 using green beans instead of Jeju green beans (fox beans).

[ test example One] bacillus fermented with isolates Jeju Green Beans According to the treatment of the derived extract mammary gland Confirmation of the effect of increasing cell proliferation of cells

Since dermal papilla cells, which play a key role in hair production, exhibit effects of preventing hair loss and/or improving hair growth through proliferation, it was confirmed whether the extract of Jeju blue bean fermented with a Bacillus sp. strain affects the proliferation of dermal papilla cells.

First, dermal papilla cells (iDPC; immortalized Dermal Papilla Cells) were established through immortalization of cells obtained by isolating the dermal papilla from human hair follicles and culturing them.

Then, the established dermal papilla cells were transferred to each well of a 96-well microplate (96-well microplate, Nunc, Wiesbaden, Germany) using DMEM (Dulbecco's Modified Eagle's Medium) containing 10% FBS (Fetal Bovine Serum, Gibco). After seeding (seeding) at 2,000 cells / well (well), 5% CO ₂ /37 ° C incubator for about 24 hours. After culturing, removing the DMEM containing 10% FBS of the 96-well microplate used for dermal papilla cell culture, and containing the extract according to Example 1, Comparative Example 1, Comparative Example 2 or Comparative Example 3 at a final concentration of 20ppm After the addition of SF-DMEM (serum free-DMEM), it _{was cultured in an incubator at 5% CO 2} /37° C. for about 48 hours ([negative control group: SF-DMEM treatment group]; [positive control group: DMEM treatment containing 5% FBS] group]). After culturing, the proliferation rate of the dermal papilla cells treated in Example 1 and Comparative Examples 1 to 3 was measured using the CCK-8 Kit to confirm the cell proliferation rate.

As a result, as shown in FIG. 1, compared to the negative control group, the Jeju green bean-derived extract fermented with the Bacillus sp. strain according to Example 1 increased the cell proliferation of dermal papilla cells by 54%, in Comparative Examples 1 to 3 It was confirmed that the cell proliferation was higher than that of the extract derived from common soybeans (soybean, seoritae, or yak bean) fermented with the Bacillus sp.

[ test example 2] bacillus fermented with isolates Jeju Green Beans Confirmation of changes in growth factors of dermal papilla cells according to the treatment of the derived extract

The effect of preventing hair loss and/or improving hair growth was confirmed by checking the expression levels of VEGF and HGF, known as growth factors involved in the proliferation of dermal papilla cells (Gnann et al., BMC Dermatol , 13:15, 2013; Yamazaki et al. . , J Investig Dermatol Symp Proc . , 4(3):312-5, 1999).

The dermal papilla cells established in Test Example 1 were seeded at 25,000 cells/well in each well of a 6-well plate (6-well plate, Nunc, Wiesbaden, Germany) using DMEM containing 10% FBS, and then approximately Incubated in a 5% CO ₂ /37° C. incubator for 24 hours. After culturing, removing the DMEM containing 10% FBS of the 6-well microplate used for dermal papilla cell culture, and containing the extract according to Example 1, Comparative Example 1, Comparative Example 2 or Comparative Example 3 at a final concentration of 20ppm After the addition of SF-DMEM (serum free-DMEM), it _{was cultured in an incubator at 5% CO 2} /37° C. for about 48 hours ([negative control: SF-DMEM treated group]; [positive control: minoxidil-containing DMEM treated group] ). After culture, the expression levels of each of VEGF and HGF in the dermal papilla cells treated with Example 1 were confirmed with the Human VEGF Quantikine ELISA Kit and the Human HGF Quantikine ELISA Kit (R&D Systems, USA).

As a result, as shown in Figure 2, in comparison with the negative control group, the extract of Jeju green beans fermented with the Bacillus sp. strain according to Example 1 increased the VEGF expression level of the dermal papilla cells by 33%, and the VEGF of minoxidil as a positive control group It was confirmed that the effect of preventing hair loss and / or improving hair growth was excellent due to the proliferation of dermal papilla cells through the increase of the VEGF expression level in the dermal papilla cells.

In addition, as shown in FIG. 3, compared to the negative control group, the extract of Jeju green beans fermented with the Bacillus sp. strain according to Example 1 increased the HGF expression level of dermal papilla cells by 23%, and the HGF release of minoxidil, a positive control group, was increased by 23%. It was confirmed that the effect of preventing hair loss and / or improving hair growth was excellent by the proliferation of dermal papilla cells through the increase of HGF expression level in dermal papilla cells.

Therefore, it was found that the extract of Jeju green beans fermented with Bacillus sp. strain had a hair growth promoting effect by promoting the expression of VEGF and HGF, which are hair growth promoting signals in dermal papilla cells.

[ test example 3] bacillus fermented with isolates Jeju Green Beans Confirmation of hair growth effect of human hair follicle organ according to the treatment of the derived extract

Human hair follicle organs were isolated one by one from the tissue remaining after hair transplantation, and William' E medium (L-glutamine (2 mM), insulin (10 μg/mL), hydrocortisone (40 ng/mL), antibiotics (1% w/v), antifungal agents (1% w/v) containing medium). After the extract according to Example 1 was treated at a concentration of 50 ppm ([negative control group: DMEM]; [positive control group: 1 μM minoxidil]), the length of hair grown from the hair follicle organ was measured on the 6th day of culture.

As a result, as shown in FIG. 4, the extract according to Example 1 exhibited a hair growth effect equal to or greater than that of the positive control group at a concentration of 50 ppm.

Therefore, it was confirmed that the extract of Jeju green beans fermented with Bacillus sp. strain was effective in growing human hair follicle organ.

The pharmaceutical, food and cosmetic composition containing the extract of the fermented Jeju green beans of the present invention, preferably the extract of Jeju green beans fermented with a Bacillus sp. strain as an active ingredient, is commercialized in various formulations of Formulation Examples 1 to 10 (pharmaceutical, food , cosmetics), but is not limited thereto, considering the functionality, cost and other conditions of the product to be implemented, an extract of fermented Jeju green beans in an appropriate amount, preferably an extract of Jeju green beans fermented with a Bacillus sp. It can be prepared into the formulation of various formulations and formulations containing, such as pharmaceutical, food and cosmetic compositions.

[Formulation Example 1] Preparation of soft capsules

An extract of fermented Jeju green beans (preferably, an extract of Jeju green beans fermented with a Bacillus sp. strain) 80 mg, vitamin E 9 mg, vitamin C 9 mg, palm oil 2 mg, hydrogenated vegetable oil 8 mg, yellow wax 4 mg and lecithin 9 mg were mixed, A soft capsule filling solution was prepared by mixing according to the method. Soft capsules were prepared by filling 400 mg per capsule. Separately, a soft capsule sheet was prepared in a ratio of 66 parts by weight of gelatin, 24 parts by weight of glycerin, and 10 parts by weight of sorbitol solution, and the filling solution was filled to prepare a soft capsule containing 400 mg of the composition according to the present invention.

[Formulation Example 2] Preparation of tablets

An extract of fermented Jeju green beans (preferably, an extract of Jeju green beans fermented with Bacillus sp. strain) 80 mg, vitamin E 9 mg, vitamin C 9 mg, galactooligosaccharide 200 mg, lactose 60 mg and maltose 140 mg were mixed and dried in a fluidized bed dryer. After granulation using , 6 mg of sugar ester was added. 504 mg of these compositions were compressed in a conventional manner to prepare tablets.

[Formulation Example 3] Preparation of drink preparation

After mixing 80 mg of fermented Jeju green soybean extract (preferably, extract of Jeju green soybean fermented with Bacillus sp. strain), 9 mg of vitamin E, 9 mg of vitamin C, 10 g of glucose, 0.6 g of citric acid, and 25 g of liquid oligosaccharide, 300 ml of purified water was added and each bottle was filled to a volume of 200 ml. After filling the bottle, it was sterilized at 130° C. for 4 to 5 seconds to prepare a beverage.

[Formulation Example 4] Preparation of granules

An extract of fermented Jeju green beans (preferably, an extract of Jeju green beans fermented with a Bacillus sp. strain) 80 mg, vitamin E 9 mg, vitamin C 9 mg, anhydrous crystalline glucose 250 mg, and starch 550 mg were mixed, and a fluidized bed granulator was used. It was manufactured by molding into granules and then filling the fabric.

[Formulation Example 5] Preparation of nutrient lotion (milk lotion)

Nutrient lotion containing the extract of the fermented Jeju green beans of the present invention (preferably, the extract of Jeju green beans fermented with a Bacillus sp. strain) of the present invention was prepared by mixing the ingredients of Table 1 including the oily component and the aqueous component.

Ingredients Content (wt%) Purified water remaining amount Extract of fermented Jeju green beans 0.1 beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 liquid paraffin 0.5 Montana 202 (Manufacturer: Seppic) 5.0 glycerin 3.0 butylene glycol 3.0 propylene glycol 3.0 Carboxyvinyl Polymer 0.1 triethanolamine 0.2 Preservatives, Colors, Flavors appropriate amount

[ Formulation example 6] Preparation of massage cream

A massage cream containing an extract of fermented Jeju green beans of the present invention (preferably, an extract of Jeju green beans fermented with a Bacillus sp. strain) of the present invention was prepared by mixing the ingredients in Table 2 including the oily component and the aqueous component.

Ingredients Content (wt%) Purified water remaining amount Extract of fermented Jeju green beans 0.1 beeswax 10.0 Polysorbate 60 1.5 sebum 60 hydrogenated castor oil 2.0 Sorbitan sesquioleate 0.8 liquid paraffin 40.0 squalane 5.0 Montana 202 (Manufacturer: Seppic) 4.0 glycerin 5.0 butylene glycol 3.0 propylene glycol 3.0 triethanolamine 0.2 Preservatives, Colors, Flavors appropriate amount

[ Formulation example 7] Manufacture of the pack

A pack containing the extract of the fermented Jeju green beans of the present invention (preferably, the extract of Jeju green beans fermented with a Bacillus sp. strain) was prepared by mixing the ingredients in Table 3 including the oil phase component and the aqueous phase component.

Ingredients Content (wt%) Purified water remaining amount Extract of fermented Jeju green beans 0.1 polyvinyl alcohol 13.0 Sodium Carboxymethyl Cellulose 0.2 glycerin 5.0 allantoin 0.1 ethanol 6.0 PEG-12 Nonylphenyl Ether 0.3 Polysorbate 60 0.3 Preservatives, Colors, Flavors appropriate amount

[ Formulation example 8] Preparation of hair tonic

A hair tonic containing the extract of the fermented Jeju green beans of the present invention (preferably, the extract of Jeju green beans fermented with a Bacillus sp. strain) was prepared by mixing the ingredients in Table 4 including the oil phase component and the aqueous phase component.

Ingredients Content (wt%) Purified water remaining amount Extract of fermented Jeju green beans 5.0 castor oil 5.0 resorcinol 0.1 menthol 0.05 panthenol 0.2 salicylic acid 0.1 tocopheryl acetate 0.1 pyridoxine hydrochloride 0.1 ethanol appropriate amount Preservatives, Colors, Flavors appropriate amount

[ Formulation example 9] of hair shampoo Produce

A hair shampoo containing the extract of the fermented Jeju green beans of the present invention (preferably, the extract of Jeju green beans fermented with a Bacillus sp. strain) of the present invention was prepared by mixing the ingredients in Table 5 including the oil phase component and the aqueous phase component.

Ingredients Content (wt%) Purified water remaining amount Extract of fermented Jeju green beans 5.0 Ammonium Lauryl Sulfate (ALS) 20 Ammonium Laureth Sulfate (ALES) 20 cocamide mipa appropriate amount Disodium EDIT 0.03 Carbomer appropriate amount ethanol appropriate amount cocamide propyl betaine 6.0 Dimethicone (50%) 2.0 Dimethicone (70%) 2.0 DL-Panthenol appropriate amount Tocopheryl Acetate appropriate amount triethanolamine appropriate amount menthol appropriate amount Methylchloroisothiazolinone Methylisothiazolinone Mixture (MC/IMI) 0.03 methylparaben appropriate amount Preservatives, Colors, Flavors appropriate amount

[ Formulation example 10] of hair conditioner Produce

A hair conditioner containing an extract of fermented Jeju green beans of the present invention (preferably, an extract of Jeju green beans fermented with a Bacillus sp. strain) of the present invention was prepared by mixing the ingredients in Table 6 including the oil phase component and the aqueous phase component.

Ingredients Content (wt%) Purified water remaining amount Extract of fermented Jeju green beans 5.0 ethanol 3.5 Self-emulsifying glycerin monostearate 1.5 propylene glycol 2.5 Stearylmethylbenzyl ammonium chloride (25%) 7.0 methyl paraoxybenzoate 0.3 Preservatives, Colors, Flavors appropriate amount

Statistical processing

For statistical analysis, the t-test test method was used to test the significance between the two groups, and compared with the negative control group, * P <0.05, ** P <0.01, *** P <0.001 was considered a valid difference.

As described above in detail a specific part of the content of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (11)

A composition for preventing hair loss or improving hair growth, comprising an extract of fermented Jeju green beans as an active ingredient. The composition for preventing hair loss or improving hair growth according to claim 1, wherein the fermented Jeju green beans are fermented with a Bacillus sp. strain. The method of claim 2, wherein the Bacillus strain is Bacillus subtilis (Bacillus subtilis), Bacillus cereus (Bacillus cereus), Bacillus NATO (Bacillus natto), Bacillus Lee Kenny formate miss (Bacillus licheniformis), Bacillus buffer milreoseu (Bacillus pumilus ) and Bacillus thuringiensis ( Bacillus thuringiensis ) A composition for preventing hair loss or improving hair growth, characterized in that at least one selected from the group consisting of. The composition for preventing hair loss or improving hair growth according to claim 1, wherein the content of the extract is 0.1 to 20.0% by weight based on the total weight of the composition. The composition for preventing hair loss or improving hair growth according to claim 1, wherein the extract promotes the expression of VEGF (Vascular Epidermal Growth Factor) and HGF (Hepatocyte Growth Factor) of dermal papilla cells. The composition for preventing hair loss or improving hair growth according to claim 5, wherein the extract promotes the expression of VEGF and HGF in dermal papilla cells at 1-50 ppm based on ppm (w/w). The composition for preventing hair loss or improving hair growth according to any one of claims 1 to 6, wherein the composition is a pharmaceutical composition, a health functional food composition or a cosmetic composition. A method for producing fermented Jeju green bean extract,
(a) inoculating the Bacillus sp. strain into Jeju green beans and then fermenting them to obtain a fermented product of Jeju green beans;
(b) preparing a reactant by adding a solvent to the fermented product and then drying the filtrate of the reaction product obtained by incubation to obtain dry matter 1 by filtration;
(c) preparing a reaction product by adding a solvent to Jeju green soybeans and drying the filtrate of the reaction product obtained by leaving it to stand to obtain dry matter 2 (dry matter 2); and
(d) mixing the filtered dried product 1 and the filtered dried product 2 to obtain a fermented Jeju green bean extract;
The method for producing a fermented Jeju green bean extract, characterized in that the content of each of the Jeju green beans of step (a) and the Jeju green beans of step (c) is the same. The method of claim 8, wherein the Bacillus strain is Bacillus subtilis (Bacillus subtilis), Bacillus cereus (Bacillus cereus), Bacillus NATO (Bacillus natto), Bacillus Lee Kenny formate miss (Bacillus licheniformis), Bacillus buffer milreoseu (Bacillus pumilus ) and Bacillus thuringiensis ) A method for producing a fermented Jeju green bean extract, characterized in that at least one selected from the group consisting of. The method of claim 8, wherein the solvent in steps (b) and (c) is one selected from the group consisting of water, an organic solvent, and mixtures thereof. The fermented Jeju green beans according to claim 8, wherein the Jeju green bean extract fermented in step (d) is obtained by mixing the filtered dried product 1 and the filtered dried product 2 in a ratio of 1 to 9: 9 to 1 by weight. Method for preparing the extract.

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