KR101985356B1 - Composition for skin external application containing fermented soybean extract - Google Patents

Abstract

The present invention provides an external preparation for skin comprising a fermented soybean extract as an active ingredient. According to the composition for external application for skin containing the fermented soybean extract of the present invention, the content of the fermented soybean extract promotes skin cell regeneration, wound healing, prevention and improvement of skin troubles such as atopy, hair loss prevention effect, It is possible to provide a skin external composition composition and a hair composition for various purposes as cosmetics or medicaments by providing an improvement effect of gloss and softness, a hair growth effect, and a hair loss prevention effect.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for skin external application containing a fermented soybean extract,

The present invention relates to a composition for external application for skin containing fermented soybean extract, and more particularly to a composition for external application for skin, which promotes skin cell regeneration, promote wound healing, improve atopy, prevent hair loss, prevent dandruff, improve hair gloss and softness, Of the present invention.

Skin is the primary barrier of the human body and protects the internal organs of the body from changes in temperature and humidity, from external stimuli such as ultraviolet rays and pollutants, and plays an important role in body homeostasis such as body temperature control. However, excessive physical and chemical stimuli from outside, stress and nutritional deficiency deteriorate the normal function of the skin and promote skin aging phenomenon such as loss of elasticity, keratinization and wrinkle formation. To prevent such phenomenon, There has been an effort to maintain skin's natural functions and activate skin cells to effectively inhibit skin aging by using cosmetics having physiologically active substances obtained from various plants, microorganisms, However, most conventional cosmetic raw materials have various problems such as insufficient efficacy and skin side effects.

In addition, as the tendency to emphasize beauty has recently increased, many people, both male and female, use hair dye or permanent agent to satisfy the need for individual expression. However, in some sensitive scalp, chemical treatment such as dyeing and perming may cause stimulation and inflammation such as itching or stinging of the scalp, and such stimulation and inflammation lead to environmental hair loss. In addition, the hair and scalp are part of the body, and as the skin ages, the skin ages like a skin, resulting in various changes and problems.

Therefore, it is possible to reduce the irritation and inflammation of the scalp caused by the use of a composition for external application for skin having excellent efficacy and skin stability and a variety of hair dye and permanent agents, to protect hair from harmful environments, to strengthen hair follicles, prevent hair loss, There is a need for a composition capable of inhibiting hair growth and promoting hair growth.

Published Patent No. 10-2010-0117514

In order to solve the above-mentioned problems, the present invention provides a fermented soybean extract, which is obtained by fermenting soybean, fermenting an extract obtained from soybean, or fermented soybean extract as a mixture thereof as an active ingredient, It is an object of the present invention to provide a composition for external application for skin.

In order to solve the above-mentioned problems, the present invention provides a composition for external application for skin, which comprises a fermented soybean extract as an active ingredient.

In one embodiment of the present invention, the fermented soybean extract may be extracted after fermentation of soybean, fermented with an extract obtained from soybean, or a mixture thereof.

In one embodiment of the present invention, the fermentation of the fermented soybean extract may be one or more selected from the group consisting of natural fermentation, bacterial fermentation and salting fermentation.

In one embodiment of the present invention, the fermented soybean extract may be extracted from water, glycerin, propanediol, lower alcohols having 1 to 4 carbon atoms, or a mixed solvent thereof at 10 to 90 ° C for 4 to 24 hours.

In one embodiment of the present invention, the fermented soybean extract may be contained in an amount of 0.01 to 99% by weight based on the total weight of the composition.

In one embodiment of the present invention, the composition may be effective for promoting skin cell regeneration, promoting wound healing, improving atopy, preventing hair loss, preventing dandruff, improving gloss and softness of hair, preventing hair growth and preventing hair loss.

In one embodiment of the present invention, the composition may have an effect of stimulating skin cell regeneration by collagenase production inhibition.

In one embodiment of the present invention, the composition may be one which promotes skin cell regeneration by promoting cell proliferation, prevents dandruff and hair growth.

In one embodiment of the present invention, the composition may have the effect of inhibiting the occurrence of black cohosh and prevention of black cohosh by promoting melanin production.

In one embodiment of the present invention, the composition may be one that promotes MITF and tyrosinase expression.

According to the composition for external application for skin containing the fermented soybean extract of the present invention, the content of the fermented soybean extract promotes skin cell regeneration, wound healing, prevention and improvement of skin troubles such as atopy, hair loss prevention effect, It is possible to provide a skin external composition composition and a hair composition for various purposes as cosmetics or medicaments by providing an improvement effect of gloss and softness, a hair growth effect, and a hair loss prevention effect.

Hereinafter, preferred embodiments of the present invention will be described in detail in order to facilitate the present invention by those skilled in the art.

The present invention provides an external preparation for skin comprising a fermented soybean extract as an active ingredient.

In the present invention, the fermented soybean extract may be obtained by fermenting soybean, fermenting the extract obtained from soybean, or a mixture thereof.

The term " fermentation " in the present invention means a process in which a microorganism decomposes an organic substance using an enzyme contained therein. In the present invention, the fermentation can be performed by a conventional fermentation method. Preferably, natural fermentation, bacterial fermentation and salting fermentation may be carried out by one or more methods. Specifically, it may be made by a microorganism generally used for fermentation, and may be made by a non-pathogenic microorganism, yeast or dry yeast.

The fermented soybean extract after fermentation of the soybean is obtained, for example, by extracting the fermented soybean after natural fermentation, bacterial fermentation or salting fermentation of the soybean. The method for preparing the extract is not particularly limited as long as it is a method known in the art. For example, water or ethanol or a mixture thereof may be added and the mixture may be extracted at 10 to 90 ° C for 4 to 24 hours.

In addition, the fermentation soybean extract obtained by fermenting the extract obtained from the soybean can be produced, for example, by extracting the soybean extract by a method known in the art and extracting the extracted soybean by natural fermentation, bacterial fermentation or salt fermentation. The soybean extract may be produced by any method known in the art without limitation, but may be, for example, extraction with water or ethanol or a mixture thereof at 10 to 90 ° C for 4 to 24 hours.

Preferably, the solid component is finely divided and then extracted with water, glycerin, propanediol, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, or a dry extract obtained by removing the solvent by various methods such as drying, vacuum distillation and freeze- Can be obtained. In addition, the extract of the present invention may be subjected to a purification process commonly used in the extraction process, for example, filtration, recrystallization, and various kinds of chromatography.

The fermented soybean extract may be contained in an amount of 0.01 to 99% by weight based on the total weight of the composition. If the content of the fermented soybean extract is less than 0.01% by weight, the effect of promoting skin cell regeneration, wound healing effect, prevention and improvement of skin troubles such as atopy, prevention of hair loss, effect of preventing dandruff, improvement of gloss and softness of hair, And the content of the formulation raw material contained in order to make the external preparation for application to the skin and the hair is not sufficient and the application easiness is poor.

The composition for external application for skin according to the present invention can increase the content of active ingredient by using a fermented soybean extract as compared with the case of using a soybean extract and has various excellent effects on skin, scalp, and hair. The composition for external application for skin of the present invention may be one having an effect of promoting skin cell regeneration, promoting wound healing, improving atopy, preventing hair loss, preventing dandruff, improving gloss and softness of hair, preventing hair growth and preventing hair loss.

The composition for external application for skin according to the present invention may have an effect of promoting skin cell regeneration by inhibition of collagenase production. Thus, skin aging can be suppressed. This can be confirmed in a test example to be described later.

In addition, the composition for external application for skin according to the present invention may be one having the promotion of skin cell regeneration by promoting cell proliferation, the prevention of dandruff and the hair growth. This can be confirmed in the following test examples.

Specifically, skin regeneration can be promoted by cell proliferation of normal fibroblasts.

In addition, hair growth can be promoted by cell proliferation of dermal papilla cells (DPC). The dermal papilla cells are the cells located at the base of the hair follicle. They supply oxygen and nutrients to the cells constituting the hair follicle, and control hair growth and follicle cycle regulation. Therefore, when the growth of the dermal papilla cells is promoted, the hair becomes healthy, hair growth is promoted, and hair loss can be prevented.

In addition, dendritic cells can be reduced by cell proliferation of fibroblasts.

The composition for external application for skin according to the present invention may be one which has the effect of preventing melanosis and preventing white hair by promoting the production of melanin and promoting the expression of MITF and tyrosinase. This can be confirmed in the following test examples.

The composition for external application for skin according to the present invention is a generic term that can be applied to any part of the skin, for example, skin, scalp or hair, and various forms of cosmetics and medicines may be included therein. That is, the external preparation for skin according to the present invention may be, for example, a pharmaceutical composition or a cosmetic composition. Formulations of the composition for external application for skin may be in the form of cosmetics, gels, water-soluble liquids, creams, essences, basic cosmetic formulations comprising emulsions and ointments of O / W type and W / O type, But are not limited to, color cosmetic formulations comprising a foundation, a skin cover, a lipstick, a lip gloss, a face powder, a two-way cake, an eye shadow, a teak color and an eyebrow pencil.

The pharmaceutical composition may further contain a preservative, a stabilizer, a wetting agent or an emulsifying accelerator, a pharmaceutical adjuvant such as a salt and / or a buffer for controlling osmotic pressure, and other therapeutically useful substances, May be formulated into oral administration forms. Parenteral dosage forms, and may be, for example, but not limited to lotions, ointments, gels, creams, patches or spray formulations.

The dosage of the active ingredient is within the level of those skilled in the art, and the daily dose of the drug depends on various factors such as the degree of progress of the subject to be administered, the age of onset, age, health condition, As a standard, it is generally possible to administer the composition at a dose of 1 / / kg to 200 mg / kg, preferably 50 / / kg to 50 mg / kg, once or twice a day, Are not intended to limit the scope of the present invention.

The formulation of the cosmetic composition is not particularly limited and may be appropriately selected according to the purpose. For example, it can be applied to softeners (skin lotion and milk lotion), nutrition lotion, essence, nutrition cream, massage cream, pack, gel, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, A body lotion, a body cream, a body oil, and a body essence, but the present invention is not limited thereto.

The content of the active ingredient is not particularly limited, but may be 0.01 to 99% by weight, preferably 0.01 to 20% by weight based on the total weight of the composition. When the active ingredient satisfies the above content, it can exhibit excellent efficacy without side effects.

The cosmetic composition may also comprise a cosmetically acceptable excipient which acts as a diluent, dispersant or carrier so as to facilitate distribution of the composition when applied to the skin. Preferably, the composition is an oil-in-water (O / W) emulsion, and the emulsion preferably contains water in an amount of at least 80% by weight of the excipient. However, the present invention is not limited thereto, and any known excipient for cosmetics may be used.

The cosmetic composition may also contain cosmetic auxiliaries commonly used in the field of the present invention such as lipids, organic solvents, silicones, thickeners, softeners, supplemental sunscreens, foam inhibitors, moisturizers, perfumes, preservatives, Pigments or nanopigments (in particular, ultraviolet radiation may be applied physically or chemically) to the surface of the substrate, such as, but not limited to, water, a total charge, a sequestering agent, a blocking agent, a cationic, anionic, nonionic or ampholytic polymers or mixtures thereof, propellants, To provide a photoprotective effect supplementarily), or various other ingredients commonly used in cosmetics, especially those used in the manufacture of sunscreen compositions.

Among the organic solvents, lower alcohols and polyols, such as ethanol, isopropanol, propylene glycol, glycerol, sorbitol and the like can be mentioned.

The fatty material may comprise oil or wax or mixtures thereof, fatty acids, fatty acid esters, fatty alcohols, petrolatum, paraffin, lanolin, hydrogenated lanolin or acetylated lanolin. Oils may be selected from animal, vegetable, mineral or synthetic oils, especially hydrogenated palm oil, hydrogenated castor oil, liquid petrolatum, liquid paraffin, purcellin oil, volatile or nonvolatile silicone oil and isoparaffin.

The composition for hair in the composition for external application for skin according to the present invention can be used before, after, before or after hair dyeing or bleaching, during or after a wave permor or straight permer, for lotion or gel for hair styling or treatment, A lotion or gel for a setter, a hair lacquer or a composition for a wave or straight permanent composition of hair, and a composition for hair dyeing or discoloring.

The composition for hair of the present invention is not particularly limited in formulation, but may be formulated into hair tonic, hair nutritional lotion, scarf treatment, hair treatment, hair rinse, hair shampoo, hair lotion or scalp hair- .

In addition, in the respective formulations, the ingredients other than the above-mentioned essential ingredients can be suitably selected and blended by those skilled in the art depending on the kind of the external preparation, the purpose of use, and the like. That is, components commonly used in conventional products can be added to the composition within a range not to impair the effect of the present invention. For example, oils, fragrances, thickeners, dispersants, solubilizers, surfactants and the like may be used, but they are not particularly limited depending on their use.

The present invention will be described in more detail through the following examples. However, the examples are for illustrating the present invention, and the scope of the present invention is not limited thereto.

[Example 1] Preparation of fermented soybean extract

1.5 kg of soybeans were added to 2 liters of water and boiled at medium temperature (60 to 80 ° C) for 1 hour. After boiling the beans, the beans were cooled at room temperature for 1 hour. The beans were put in a pot and stored in a shade for 3 days at room temperature. Then, the beans were pulverized with a blender, and then 7 liters of an 80% aqueous ethanol solution was added, refluxed three times, and then immersed at 15 ° C for 1 day. Thereafter, the residue and the filtrate were separated by filtration and centrifugation through a filter cloth, and the separated filtrate was concentrated under reduced pressure to obtain 421 g of a fermented soybean extract.

[Comparative Example 1] Production of soybean extract

1.5 kg of soybean was pulverized with a mixer, and 7 liters of 80% aqueous ethanol solution was added thereto, refluxed three times, and then immersed at 15 ° C for 1 day. Thereafter, the residue and the filtrate were separated by filtration through a filter cloth and centrifugation, and the separated filtrate was concentrated under reduced pressure to obtain 343 g of a soybean extract.

[Formulation Example 1 and Comparative Formulation Examples 1 and 2]

In order to use the composition of the present invention as a cosmetic, a cream containing a fermented soybean extract and a soybean extract was prepared in the composition shown in Table 1 below.

Ingredients (% by weight) Formulation Example 1 Comparative Formulation Example 1 Comparative Formulation Example 2 Purified water To 100 To 100 To 100 Fermented soybean extract 0.1 - - Soybean extract - 0.1 - Vegetable hydrogenated oil 1.50 1.50 1.50 Stearic acid 0.60 0.60 0.60 Stearyl alcohol 2.00 2.00 2.00 Glycerol stearate 1.00 1.00 1.00 Polyglyceryl-10 < / RTI > pentastearate &
Behenyl alcohol and sodium stearoyl lactylate 1.00 1.00 1.00 Arachidyl behenyl alcohol and arachidyl glucoside 1.00 1.00 1.00 Cetylaryl Alcohol and Cetearyl Glucoside 2.00 2.00 2.00 PEG-100 Stearate & Glycerololate & Propylene Glycol 1.50 1.50 1.50 Caprylic / capric triglyceride 11.00 11.00 11.00 Cyclomedicone 6.00 6.00 6.00 Triethanolamine 0.1 0.1 0.1

[Test Example 1] Test for improving atopy symptoms in the human body

In order to confirm the effect of improving the atopic symptoms by the composition of the present invention, the cream prepared in Formulation Example 1 and Comparative Formulation Examples 1 and 2 was applied to the panels to evaluate the degree of improvement of atopic dermatitis. For each panel of 15 panelists who thought that they had symptoms of dermatitis showing similar symptoms, each composition was divided into 3 groups of 5 each, and each composition of the following Table 2 was administered twice daily for 12 weeks at a temperature of 24 to 26 캜 and a humidity of 75% And applied to the face. At the end of the test, subjects were asked to fill out a questionnaire and conducted a subjective efficacy evaluation. In the questionnaire, symptoms of atopic dermatitis were classified as "pruritus", "dryness", "keratinization", "dandruff", "erythema", "swelling", "skin crack", " And the degree of improvement was evaluated for each symptom in the case of the above symptoms, and the improvement effect of the atopy symptom was evaluated on an individual basis. The results are shown in Table 2.

Application group (number of respondents - persons) Formulation Example 1 Comparative Formulation Example 1 Comparative Formulation Example 2 Significant improvement 4 One Improving One 2 No change 2 5 worse

From the above Table 2, it can be confirmed that the composition containing the fermented soybean extract of the present invention further improved the symptoms of the atopic dermatitis as compared with the Comparative Formulation Example 2. [ In addition, it was confirmed that the fermented soybean extract further improved the symptoms of the atopic dermatitis than the case of applying the composition containing the soybean extract.

[Test Example 2] Collagenase inhibitory effect

The ability of the fermented soybean extract prepared in Example 1 to inhibit collagenase production was measured in comparison with soybean extract (Comparative Example 1), tocopherol (Comparative Example 2) and EGCG (Comparative Example 3). Tocopherol and EGCG are substances known to regenerate epidermal cells of the skin and prevent aging of the skin.

Human fibroblasts were plated at 5,000 cells / well in a 96-well microtiter plate containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% fetal bovine serum, Lt; / RTI > until growth. Then, the cells were cultured in a serum-free DMEM medium for 24 hours. Then, 100 μg / ml concentration of the fermented soybean extract of Example 1, 100 μg / ml concentration of the soybean extract of Comparative Example 1, 10 g / ml of tocopherol and EGCG dissolved in serum- ^-4 molar ratio for 24 hours, and then the cell culture solution was collected.

The degree of collagenase production in the cell culture was measured using a collagenase measuring device (Amersham Pharmacia, USA). First, the cell culture solution collected in a 96-well plate uniformly coated with the primary collagenase antibody was added and the antigen-antibody reaction was performed in a thermostatic chamber for 3 hours.

After 3 hours, the second collagen antibody bound to the chromophore was put into a 96-well plate and reacted for another 15 minutes. After 15 minutes, the color development inducing substance was added, coloring was performed at room temperature for 15 minutes, and 1 M sulfuric acid was added to stop the reaction (color development). The color of the reaction solution became yellow, and the degree of yellow was different depending on the progress of the reaction Respectively.

The absorbance of a yellow 96-well plate was measured at 405 nm using a spectrophotometer, and the degree of synthesis of collagenase was calculated by the following equation (1). At this time, the reaction absorbance of the cell culture obtained from the untreated group was used as a control. That is, the degree of expression of collagenase in the untreated group was set at 100, and the degree of expression of collagenase in the group treated with the test substance was determined, and the results are shown in Table 3 below.

[Equation 1]

Collagenase expression level (%) = A / B x 100

A: Absorbance of the test substance-treated cell group

B: Absorbance of the control group

Test substance Expression level of collagenase (%) Untreated group 100 Soybean extract (Comparative Example 1) 86 Tocopherol (Comparative Example 2) 76 EGCG (Comparative Example 3) 59 Fermented soybean extract 61

As can be seen from Table 3, the fermented soybean extract effectively inhibited the expression of collagenase in vitro and was superior to tocopherol, which is known as an antioxidant, in suppressing the expression of collagenase . In addition, the fermented soybean extract was found to be superior to the soybean extract in inhibiting the expression of collagenase.

[Test Example 3] Effect of increasing cell proliferation

Human skin fibroblasts were inoculated into each well of a 96-well microplate so as to have a skin cell reproduction effect as a skin regeneration effect so as to be 1 × 10 ⁴ cells. And cultured in DMEM (Dulbecco's Modified Eagle's Medium) for 24 hours. After fermentation, the fermented soybean extract and soybean extract were replaced with serum-free DMEM medium adjusted to a final concentration of 100 μg / ml, and further cultured for 24 hours. 3- (4,5-Dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium (MTT: 5 mg / ml) was added to each well and allowed to stand for 4 hours, after which the medium was discarded. 100 μl of dimethyl sulfoxide solution was added to each well and the absorbance was measured at 570 nm with a microplate reader.

The above procedure was tested to determine the cell proliferation effect from the following equation (2), and the results are shown in Table 4 below.

&Quot; (2) "

Cell proliferation effect (%) =

Cell regeneration effect Test substance Absorbance (570 nm) Cell proliferation effect (%) Control (μg / ml) 0.300 - Fermented soybean extract (100 μg / ml) 0.475 58.3 Soybean extract (100 μg / ml) 0.321 7.0

As shown in Table 4, the fermented soybean extract showed cell proliferation effect of normal fibroblasts. In addition, the fermented soybean extract has a better cell proliferation effect than the soybean extract.

 [Formulation Example 2 and Comparative Formulation Examples 3 and 4]

Shampoos were prepared with the compositions shown in Table 5 below. Specifically, the surfactant and ethylene glycol distearate were added to purified water, heated to 80 DEG C to dissolve uniformly, and gradually cooled to 40 DEG C with stirring. To the mixture was added the extract of the present invention, preservative, A viscosity adjusting agent, a perfume, and a hair conditioning agent were mixed and stirred, followed by cooling to room temperature.

ingredient Formulation Example 2 Comparative Formulation Example 3 Comparative Formulation Example 4 Ammonium lauryl sulfate 10 10 10 Polyoxyethylene lauryl ammonium sulfate 5 5 5 Cocoamidopropyl betaine 2 2 2 Ethylene glycol distearate 1.5 1.5 1.5 Cocoyl monoethanolamide 0.8 0.8 0.8 Fermented soybean extract 5.0 - - Soybean extract - 5.0 - Polyquaternium-10 0.2 0.2 0.2 Blue No. 1 0.0002 0.0002 0.0002 Yellow No. 4 0.0001 0.0001 0.0001 Methyl paraben 0.1 0.1 0.1 Spices 0.8 0.8 0.8 Citric acid 0.1 0.1 0.1 Dimethicone 1.0 1.0 1.0 water to 100 to 100 to 100

[Test Example 4] Anti-dandruff evaluation

Anti-dandruff power of Formulation Example 2 and Comparative Formulation Examples 3 and 4 was measured and compared. At this time, as a test strain, dicembryum Pityrosporum Ovalae (ATCC 12078) was used. The skin disc diffusion method was used to measure the antibacterial activity. The skin disc diffusion method (SDDM) is the method closest to the consumer's habits and is a method of measuring the antimicrobial activity against guinea pig skin similar to human skin. Specifically, guinea pig skin was peeled off, treated with 70% ethanol, uniformly spread, and skin disks cut to a certain size were used. The sterilized skin discs were immersed in the shampoo dilution solution for 3 minutes and then rinsed with flowing water. Then, the skin discs were placed on a solid medium inoculated with the test bacteria, and then the test bacteria were cultured. After the culture, the size of the clear zone in which the growth of the bacteria was inhibited was measured, and the relative antibacterial activity was measured. The experimental results are shown in Table 6 below as the average value of the results of three measurements.

Anti-dandruff evaluation division Restraining Size (mm) Formulation Example 2 Comparative Formulation Example 3 Comparative Formulation Example 4 Strain Reduction Size 3.2 0.5 0

Table 6 shows that Comparative Formulation Example 4, which did not contain fermented soybean extract, had no effect of reducing dandruff, but effectively reduced dandruff in Formulation Example 2 containing fermented soybean extract. In addition, it can be confirmed that the anti-dandruff effect is excellent because the effect of reducing dandruff is superior to that of Comparative Example 3 containing soybean extract.

[Test Example 5] Dandruff reduction effect test

A total of 36 men aged 19 to 35 years who had relatively high dandruff were selected and used in three groups of 12 persons for each of Formulation Example 2 and Comparative Formulation Examples 3 and 4 for 1 month in the following manner, Respectively.

Before the start of the test, the hair was smeared with a normal shampoo, and the dandruff accumulated for 2 days after shedding was collected. The dandruff weight was sampled once every two days with the shampoo of each of Formulation Example 2 and Comparative Formulation Examples 3 and 4, The weights of accumulated dandruff for two days were compared and evaluated. The accumulated dandruff was collected directly from the scalp using a vacuum suction device, and the dandruff reduction rate was calculated according to the following equation (3). The results are shown in Table 7 below.

&Quot; (3) "

Formulation Example 2 Comparative Formulation Example 3 Comparative Formulation Example 4 Dandruff reduction rate (%) 88.87 31.99 0.7 93.02 33.48 -0.5 73.21 26.35 -1.1 98.00 35.28 1.5 94.68 34.08 2.2 85.07 30.62 1.3 78.30 28.18 6.3 75.22 27.07 3.6 89.22 32.11 3.3 93.25 33.57 4.6 88.27 31.77 3.7 79.25 28.53 4.2 Dandruff reduction rate (%, average value) 86.4 31.09 2.5 SD 8.14 2.93 2.2

As can be seen from the above Table 7, Formulation Example 2 containing the fermented soybean extract shows excellent dandruff-preventing effect. In addition, it can be seen that the dandruff prevention effect is superior to Comparative Example 3 containing soybean extract.

[Test Example 6] Test for preventing the itchiness of the scalp

Twenty-four male and female twenty-four to twenty-five to forty-five years of age who felt severe scalp itching were selected and divided into three groups of eight, and the shampoo of each of Formulation Example 2 and Comparative Formulation Examples 3 and 4 was used once for three days for two weeks The scalp itching prevention effect was evaluated through the following evaluation criteria, and the results are shown in Table 8 below.

[Evaluation standard]

Very good - 5 points

Excellent - 4 points

Usually - 3 points

Poor - 2 points

Very bad - 1 point

division Formulation Example 2 Comparative Formulation Example 3 Comparative Formulation Example 4 The itching effect of scalp 4.4 2.8 2.3

As can be seen from the above Table 8, it can be seen that Formulation Example 2 containing the fermented soybean extract shows a superior effect in preventing scalp itching. In addition, it can be seen that the formulation containing the fermented soybean extract has a better effect than the Comparative Formulation Example 3 containing the soybean extract.

[Test Example 7] Effect of proliferation of dermal papilla cells

The keratin proteins that make up the hair are produced in hair follicle keratinocyte, which keratinocytes differentiate from the dermal papilla cells. In order to evaluate the activity of the dermal papilla cells of the present composition, DP6 (rat immortalized dermal papilla cell) cell line was used in the present invention (Wendy Filsell, Journal of Cell Science 107, 1761-1772 (1994)). This dermal papilla cell line was obtained by microdissection in the hair roots of male PVG rat beard and was cultured in DMEM (Dulbecco's modified Eagle's medium, Gibco BRL, Gaithersburg, MD, USA) containing 10% FBS (Fetal bovine serum) USA) for 24 hours in an incubator maintained at 37 ° C in 5% CO 2. DP6 was added to a 96-well plate and cultured in a 37 ° C incubator for 24 hours. Then, the present fermented soybean extract was treated at concentrations of 10 ppm, 50 ppm and 100 ppm, respectively. After 24 hours of drug treatment, cell proliferation was measured using WST-1 kit (Roche). The results are shown in Table 9 below.

division Cell proliferation (%) Untreated Control (Control) 100 Fermented soybean extract (10ppm) 112 Fermented soybean extract (50ppm) 127 Fermented soybean extract (100ppm) 148 Soybean extract (10ppm) 101 Soybean extract (50ppm) 103 Soybean extract (100ppm) 111

As can be seen from Table 9, when the fermented soybean extract was treated, the proliferation of the dermal papilla cells was increased, which was significantly increased depending on the concentration. In addition, the fermented soybean extract treated with soybean extract showed better proliferation of dermal papilla cells.

[Test 8] Evaluation of Potassium Ion Channel Activity Increase Effect

Minoxidil, a therapeutic agent for depilation, is known as a potential mitochondrial potassium ion channel opener (KATP channel opener) and is a representative drug used for the treatment of androgenetic alopecia. To evaluate the mechanism of minoxidil, cytotoxicity was inhibited by treatment with tolbutamide (SIGMA AlDRICH, T0891) blocking the KATP channel in fibroblasts constituting the dermis of the scalp, and the cell proliferation was restored by opening the potassium ion channel again Was used.

To evaluate the function of the present composition as a KATP channel opener, the fibroblast cell line NIH3T3 (mouse embryonic fibroblast cell line) cell line was used in the present invention. This cell line is a cell line obtained by natural immortalization of fibroblasts isolated from NIH Swiss mouse embryo by the 3T3 protocol. The cell line was cultured in DMEM (Gibco BRL, Gaithersburg, Md., USA) containing 10% FBS for 24 hours in an incubator maintained at 37 ° C in 5% CO2. NIH3T3 was added to a 96-well plate, cultured in a 37 ° C incubator for 24 hours, treated with 2.5 mM tolbutamide, and 10 minutes after the positive control, 10 μM of minoxidil, fermented soybean extract and soybean extract were added at concentrations of 10 ppm, 50 ppm and 100 ppm , And cell proliferation was measured using WST-1 kit (Roche) after 48 hours of drug treatment. The results are shown in Table 10 below.

division Cell proliferation (%) Untreated Control (Control) 100 Minoxidil 133 Fermented soybean extract (10ppm) 114 Fermented soybean extract (50ppm) 123 Fermented soybean extract (100ppm) 135 Soybean extract (10ppm) 102 Soybean extract (50ppm) 105 Soybean extract (100ppm) 115

As shown in Table 10, when the fermented soybean extract was treated, the proliferation of fibroblasts was restored and the cell proliferating ability was found to be increased depending on the concentration of the fermented soybean extract treated. When the fermented soybean extract was changed to 100 ppm It can be confirmed that the proliferation of fibroblast cells is restored to minoxidil level. In addition, the fermented soybean extract treated with soybean extract showed better proliferation effect of fibroblast.

[Formulation Example 3 and Comparative Formulation Examples 5 and 6]

A cream-based composition was prepared in a conventional manner according to the composition of Table 11 below (unit: wt%).

Compounding ingredient Formulation Example 3 Comparative Formulation Example 5 Comparative Formulation Example 6 Labrafac 23 23 23 Tween 80 10.5 10.5 10.5 Glycerol monostearate 4.0 4.0 4.0 Stearic acid 7.0 7.0 7.0 Cetyl alcohol 2.0 2.0 2.0 Propylene glycol 9.0 9.0 9.0 Glycerol monooleate 3.5 3.5 3.5 Fermented soybean extract One - - Soybean extract - One - water 40 41 41

[Test Example 9] Wool effect of fermented soybean extract

The hair of the back of the ICR mouse was depilated by a depilator, and hair was completely removed by using a depilatory cream (bit cream). Three groups except the control group (normal group) were applied to the skin once a day with 200 쨉 l of 1% DNCB to induce skin inflammation for 3 days. In order to minimize the hair growth by setting stress conditions in the skin by inducing skin inflammation by this method and to evaluate the hair regrowing effect of the composition of the present invention, the above formulation 3 and comparative formulation 5, and 6 creams were treated and the length of hair growth was measured according to the days of treatment and compared with the control group. The results are shown in Table 12 below.

Days to be treated (days) Fur Length (cm) Formulation Example 3 Treatment Comparative Formulation Example 5 Treatment Comparative Formulation Example 6 Treatment 0 0.1 0.1 0.1 6 0.2 0.1 0.1 9 0.5 0.2 0.1 12 0.9 0.3 0.1

As shown in Table 12, the length of hair of the group treated with the cream base of Formulation Example 3 showed a statistically significant difference (p < 0.01) as compared with the group treated with the cream base of Comparative Formulation Example 6, The length of the hair was increased to about 0.9 cm on the 12th day, and the cream base material of Comparative Example 6, which did not contain the fermented soybean extract, It is possible to confirm that the length is longer. In addition, it can be confirmed that the length of hair is longer than that of the group treated with the cream base of Comparative Formulation Example 5 containing soybean extract.

This suggests that fermented soybean extract promoted hair regeneration even under conditions of restoration of hair formed under skin stress. Thus, it can be seen that the fermented soybean extract has a regenerating function for hair removed under stress.

[Test Example 10] Melanin production promoting effect test of fermented soybean extract

Melanocytes (melan-a) were added to a 24-well microtiter plate at 50,000 cells / well in RPMI medium supplemented with 5% fetal bovine serum, 100 IU penicillin G and 0.2 μM TPA Respectively. Next day, the fermented soybean extract or soybean extract as a test substance was treated at a final concentration of 10 ppm or 50 ppm, and 0.1% DMSO was used as a negative control, 100 μM IBMX was used as a positive control, and the cells were incubated at 37 ° C. for 3 days Respectively. After incubation, the wells were washed with PBS, and 100 μl of 1N NaOH was added thereto, and melanin in the cells was dissolved. The absorbance of dissolved melanin was measured at 405 nm using a microplate reader. The results of comparing the melanin production promoting effect of the fermented soybean extract with the control group are shown in Table 13 below.

sample Amount of melanin synthesis (%) DMSO (0.1%) 100 IBMX (100 [mu] M) 120 Fermented soybean extract (10ppm) 110 Fermented soybean extract (50ppm) 123 Soybean extract (10ppm) 101 Soybean extract (50ppm) 109

As shown in Table 13, fermented soybean extract promotes melanin synthesis of melanocyte to increase melanin production, thereby showing excellent melanin production promoting effect. It is also found that the fermented soybean extract has a better melanin production promoting effect than the soybean extract.

[Test Example 11] Promotion of MITF and tyrosinase expression in melanocytes of fermented soybean extract

The cells were divided into 6-well microtiter plates at 500,000 cells / well using a 501mel cell line. DMSO 0.1% as a negative control, IBMX 100 μM as a positive control group, and fermented soybean extract Or soybean extract was treated at 10 ppm and cultured at 37 ° C for 24 hours, 48 hours, and 72 hours to obtain a protein. The proteins thus obtained were subjected to western blotting using MITF and tyrosinase antibodies. Protein extraction and Western blotting were routinely performed by standard methods used by those skilled in the art. After Western blotting, the result was compared with the negative control group of 100, and the results are shown in Table 14 below.

MITF Tyrosinase IBMX Fermented soybean extract Soybean extract IBMX Fermented soybean extract Soybean extract 24hr 98 99 99 160 149 101 48hr 121 110 103 149 157 106 72hr 224 123 109 298 191 110

As shown in Table 14, it can be confirmed that fermented soybean extract increases MITF and tyrosinase protein expression in melanocytes. In addition, fermented soybean extracts showed better MITF and tyrosinase protein expression than soybean extracts.

[Test Example 12] Evaluation of anti-white coat and infertility effect of in vivo fermented soybean extract using white algae-stimulated albumin mouse

Vitiligo mice (C57bl / 6-Mitfmi-vit) were purchased from The Jackson Lab, USA. The anti-whiplash effect test method using the mouse in which the white moth is promoted is as follows. Dorsal hairs of 12-week-old mice were removed by depilation. However, the width of the hair removal area was adjusted to be the same for each individual. From the day after the hair was pulled out, twice a day, the anti-whiplash material was applied on the hairless area. A mixture of EtOH: 1,3-BG: DW = 3: 2: 5 (volume ratio) was used as the vehicle of the anti-whiplash material. The carrier was used as a negative control, and a solution containing 50 mM of IBMX As a control, 2.5% of fermented soybean extract or soybean extract was added to the test group. After about 3 weeks, when the difference in antibacterial effect between the substances was visually recognized, newly-emerged hairs were collected and the amount of melanin in the hair was measured. The amount of melanin in hair was measured using the protein hydrolyzing enzyme Esperase (Novozyme). Reaction buffer was prepared by dissolving Esperase in a buffer (50 mM Tris-HCl, 5 mM DTT, pH 9.3) to a concentration of 1 NPU / ml. Five milligrams of mouse hair was added to 1 ml of reaction buffer and allowed to react for 13 hours at 37 ° C with shaking at 1,000 rpm. Then, the hair and the reaction solution were separated by an instant centrifugation. The reaction solution thus obtained is placed in a 96-well plate, and the amount of melanin in the reaction solution can be measured by measuring the absorbance at a wavelength of 405 nm. The results are shown in Table 15 below when the negative control, the positive control, and the test group material were treated with the vitiligo-stimulated vitiligo mouse model, and their effects were visually observed and measured by the melanin quantification method in the hair.

Negative control group IMBX Fermented soybean extract Soybean extract The% 100 105.9 108.2 102.4

According to the above Table 15, it can be seen that the fermented soybean extract inhibits white moths in the body of a mouse in which white moth is promoted, and increases the amount of melanin in the hair to promote the black moth induction. In addition, the fermented soybean extract was found to be superior to the soybean extract in promoting the black cohosh.

Claims (10)

Fermented soybean extract as an active ingredient,
A cosmetic composition for external application to the skin, comprising at least one of prevention of hair loss, prevention of dandruff, improvement of gloss and softness of hair, prevention of hair growth and prevention of hair loss. The method according to claim 1,
The fermented soybean extract is obtained by fermenting soybeans and then extracting them, fermenting the extracts obtained from soybeans, or a mixture thereof. The method according to claim 1,
Wherein the fermentation soybean extract is at least one selected from the group consisting of natural fermentation, bacterial fermentation and salt fermentation. The method according to claim 1,
Wherein the fermented soybean extract is extracted with water, glycerin, propanediol, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof at 10 to 90 DEG C for 4 to 24 hours. The method according to claim 1,
Wherein the fermented soybean extract is contained in an amount of 0.01 to 99% by weight based on the total weight of the composition. delete delete The method according to claim 1,
Wherein the composition has an effect of preventing dandruff and hair growth by promoting cell proliferation. The method according to claim 1,
Wherein the composition has an effect of preventing black cohosh and prevention of black curl by promoting melanin production. The method according to claim 1,
Wherein the composition promotes microphthalmia-associated transcription factor (MITF) and tyrosinase expression.