KR20200083712A - Composition containing an mixed fermentation extract of rubus coreanus, soy bean and zanthoxylum schinifolium as an active ingredient and its use - Google Patents

Abstract

The present invention relates to a cosmetic material composition containing a composite fermented extract of Rubus coreanus, soybeans, and Zanthoxylum schinifolium as an active ingredient. More specifically, the present invention relates to the cosmetic material composition including an active ingredient produced by fermenting a composite extract of Rubus coreanus, soybeans, and Zanthoxylum schinifolium having antioxidant, anti-inflammatory, and skin whitening effects. The composition produced according to the present invention and including the composite fermented extract of Rubus coreanus, soybeans, and Zanthoxylum schinifolium has excellent antioxidant, antiaging, anti-inflammatory, skin whitening, and wrinkle alleviating effects, thereby being capable of being used as a cosmetic material composition, a pharmaceutical composition, and a health food.

Description

Cosmetic composition containing the compound fermented extract of soybean, soybean, and wild pepper as an active ingredient and its use{COMPOSITION CONTAINING AN MIXED FERMENTATION EXTRACT OF RUBUS COREANUS, SOY BEAN AND ZANTHOXYLUM SCHINIFOLIUM AS AN ACTIVE INGREDIENT AND ITS USE}

The present invention relates to a cosmetic composition containing a complex fermented extract of bokbunja, soybean and sancho as an active ingredient. In more detail, the present invention relates to a cosmetic composition comprising an active ingredient prepared by fermenting a complex extract of bokbunja, soybean and sancho, which has antioxidant, anti-inflammatory, skin whitening and wrinkle improvement effects.

Researches on bioactive substances from natural products are being actively conducted, and plant resources are widely used as treatment and preventive agents or health supplements for diseases.

Skin aging includes endogenous aging, in which the structure and function of the skin are continuously reduced along with organs over time, and exogenous aging, which is a change in skin tissue due to prolonged exposure to sunlight. Through this aging process, the skin becomes dry, wrinkles are formed, and the skin loses elasticity.

The most important topics in skin aging research are free radicals and reactive oxygen species. They are naturally produced in vivo, but they are generated by pollution, solar ultraviolet rays, chemical oxidizing agents and microorganisms. The free electromagnetic or free radicals generated at this time exert oxidative stress on the skin cells, and the material produced in this process is considered to be the cause of melanin and wrinkles.

The free electrons or free radicals generated at this time exert oxidative stress on the skin cells, and the material produced in this process is considered to be the source of melanin and wrinkles.

These free radicals disintegrate the skin antioxidant barrier made of enzymatic and non-enzymatic antioxidants, oxidative modification of biomolecules, damage to skin barrier and chain breakage of connective tissue such as collagen and hyaluronic acid, and wrinkle formation by abnormal cross-linking, etc. Accelerates skin aging

As a result, in order to prevent skin aging, it is necessary to suppress the generation of excess free radicals and to effectively remove the free radicals produced. In addition, studies on the inhibition of enzymes that cause skin wrinkles and the inhibition of enzyme activity that progresses pigmentation are needed.

On the other hand, pigmentation caused by melanin production or proliferation of melanocytes can be suppressed in various ways. In general, various pathways such as inhibition of tyrosinase activity, an important enzyme for melanin synthesis, reduction of melanocyte function, reduction of melanin production through inhibition of an automatic oxidation reaction, and inhibition of inflammatory reactions such as erythema can be used.

As a result, in order to prevent skin aging, it is necessary to suppress the generation of excess free radicals and to effectively remove the generated free radicals, and to suppress the enzymes that cause wrinkles on the skin and to inhibit the activity of enzymes that undergo pigmentation. Research is needed.

In this regard, research using plants has been conducted in relation to skin aging and whitening.

Rubus coreauns is a medicinal name that refers to the fruit of a bokbunja strawberry belonging to the family Rosaceae. Bokbunja contains ingredients such as total sugar (17.3%), reduced sugar (8.6%), crude protein (10.6%), crude ash (4.5%), crude fat (3.1%) and crude fiber (3.9%), and contains amyl alcohol It contains 11 kinds of alcohols, acids containing valeric acid, 20 kinds of carbonyls including hexanal, 5 kinds of hydrocarbons including 2-heptanone, and methyl palmitate. Contains three types of ester fragrance components. In addition, since it contains a large amount of polyphenols, it has anti-cancer effects, lowering cholesterol, suppressing hypertension or arteriosclerosis, preventing the formation of lipid peroxide, preventing aging, lowering the lipid concentration in serum, and suppressing the production of neutral lipids. It has been reported to have an effect of preventing and enhancing capillary resistance.

Soybeans are traditionally used as a source of protein in Korea because about 40% of the amount of building is composed of protein and rich in essential amino acids. Considering that the amount of protein in other cereals is usually 8~12% and that of other legumes is about 20~30%, the protein composition of soybean is very high. In addition, soybean is rich in isoflavones and phytic acid, which have recently been reported to have bioactive functions, and are widely used as raw materials for processed foods worldwide.

Sancho ( Zanthoxylum Schinifolium ) is a deciduous shrub with the dicotyledonous family of the Euphorbia musculus and grows in the mountains. The height is 3m, the twigs are thorny and reddish brown. The leaves are misaligned, composed of 13-21 small leaves. The small leaves are 1-5cm long, lanceolate, narrow at both ends, and have wavy teeth on the edges, with transparent spots. Sancho is used by picking leaves and using it raw or peeled and dried before being dried or concentrated. Sancho fruit is effective for indigestion, plant, stomach and sewage, gastric expansion, vomiting, dysentery, diarrhea, nervous breakdown, coughing, and roundworm remedies. It is used in a similar way to pepper.

Cardiovascular disease treatment use of the fermentation extract of the bimolecular molecule (refer to patent document 1), hyperlipidemia treatment use of soybean fermentation extract (refer to patent document 2), or atopy skin improvement use of a composition containing wild botanical oil derived from wild herb (see patent document 3) Studies have been conducted, but studies on antioxidant, anti-aging, anti-inflammatory and whitening efficacy, or studies related to bokbunja, soybean, and herbaceous complex fermentation extract have not been conducted.

Accordingly, the present inventors completed the present invention by confirming the antioxidant, anti-inflammatory, and whitening efficacy of the complex fermentation extract by conducting a study using the complex fermentation extract of bokbunja, soybean, and green pepper.

Korean Registered Patent Publication No. 10-0862610 Korean Registered Patent Publication No. 10-0656241 Korean Registered Patent Publication No. 10-1056475

An object of the present invention is to provide a cosmetic composition containing a complex fermentation extract of bokbunja, soybean and sancho as an active ingredient.

Another object of the present invention is to provide a method for preparing an active ingredient having anti-oxidation, anti-aging, anti-inflammatory and skin whitening effects by fermenting a complex extract of bokbunja, soybean and sancho, and a cosmetic composition comprising the active ingredient .

Another object of the present invention is to provide a health food having antioxidant, anti-aging, anti-inflammatory, and skin whitening effects containing a complex fermented extract of bokbunja, soybean, and wild pepper as an active ingredient.

The present invention provides a cosmetic composition containing a complex fermentation extract of bokbunja, soybean and sancho as an active ingredient.

The cosmetic composition may have antioxidant, anti-aging, anti-inflammatory, skin whitening and wrinkle improvement effects.

The complex extract of the bokbunja, soybean and sancho can be prepared by mixing bokbunja, soybean and sancho in a weight ratio of 1:1:1. When the weight ratio of bokbunja, soybean, and pepper in the complex extract is outside the 1:1:1 range, the fermentation efficiency during fermentation is low and the anti-inflammatory activity, antioxidant activity, whitening effect and wrinkle improvement effect of the complex fermentation extract prepared by fermentation Low and inefficient.

The active ingredient may be prepared by fermenting lactic acid bacteria with a complex extract of bokbunja, soybean, and sancho, and the lactic acid bacteria may be any of the Lactobacillus strains, preferably Pediococcus strains, Preferably Pediococcus pentosaceus B1, accession number KACC 81067BP).

The fermentation may be carried out using various strains of Pediococcus strains based on a composite extract sample in a ratio of 7% by weight, Pediococcus pentosaceus B1, accession number KACC 81067BP It is preferred to use strains. When a strain other than Pediococcus pentosaceus B1 (Accession No. KACC 81067BP) is used, fermentation efficiency is low and thus inefficient. In addition, when the proportion of the strain solution is used in an amount of less than 7% by weight or more than 7% by weight, fermentation efficiency is low and thus inefficient.

The fermentation can be carried out at 37°C for 2 to 7 days. When fermenting at a temperature lower than 37°C, the fermentation efficiency is low, and when fermenting for less than 2 days or for more than 7 days, the fermentation efficiency is low, which is inefficient.

The complex fermentation extract of the active ingredient according to the present invention, bokbunja, soybean and sancho, may be included at 50 to 600 µg/mL, more preferably 100 to 500 µg/mL, most preferably It may be included at 200 to 300 μg/mL.

When the content of the complex molecule, soybean and wild-fermented fermentation extract is less than 100 µg/mL, the whitening efficacy, antioxidant activity, anti-aging activity and anti-inflammatory activity may not be exhibited, and when it exceeds 500 µg/mL, The degree of increase in effect may be negligible, and it is not economical due to strong cytotoxicity.

The cosmetic composition may be selected from the group consisting of lotion, essence, lotion, cream, pack, foundation, gel, ointment or spray. It is not necessarily limited to this.

In addition, the cosmetic composition is a fat substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an anti-aging agent, a suspending agent, a stabilizing agent, a foaming agent, in addition to the complex fermented extract of the compound of the present invention, soybean and pepper ), fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion blockers and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipids It may contain adjuvants commonly used in the cosmetic field, such as vesicles or any other ingredients commonly used in cosmetics.

The present invention provides a pharmaceutical composition having an antioxidant, anti-aging effect and anti-inflammatory effect, which contains a complex fermented extract of bokbunja, soybean and wild pepper as an active ingredient.

The pharmaceutical composition of the present invention may further include commonly used excipients, disintegrants, sweeteners, lubricants, flavoring agents, and the like, and tablets, capsules, powders, granules, suspensions, emulsions, by conventional methods, It can be formulated as a syrup, other liquid. Specifically, the pharmaceutical composition of the present invention may be formulated for oral administration, for example, tablets, troches, lozenges, water-soluble or dominant suspensions, preparation powders or granules, emulsions, hard or soft capsules, syrups or It is formulated as elixirs. Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, stearic acid for formulation into tablets and capsules Contains lubricants such as magnesium, calcium stearate, sodium stearyl fumarate, or polyethylene glycol wax. In the case of capsule formulations, in addition to the above-mentioned substances, a liquid carrier such as fatty oil is contained.

In addition, the pharmaceutical composition of the present invention can be administered orally or parenterally, it is preferable to select a subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method when parenteral administration. In order to formulate a formulation for parenteral administration, the centifedgrass extract of the present invention, the fraction or its active fraction separated from the fraction is mixed with water with a stabilizer or buffer in a suspension to prepare a suspension and prepared as a unit dosage form of ampoules or vials do.

The dosage of the active ingredient according to the present invention is appropriately selected according to the absorption of the active ingredient in the body, the rate of inactivation and excretion rate, the patient's age, sex and condition, and severity of the disease to be treated, but in the case of oral dosage forms, in general The extract of the present invention per kg of body weight per day can be administered to an adult once or several times in an amount of 0.0001 to 500 mg, preferably in an amount of 0.001 to 100 mg.

In addition, the present invention provides a healthy food having antioxidant, anti-aging, anti-inflammatory, skin whitening and wrinkle improvement effects containing the complex fermented extract of bokbunja, soybean and wild pepper as an active ingredient.

The compound of the present invention, the complex fermentation extract of soybean and wild pepper, has excellent antioxidant, anti-aging, anti-inflammatory, skin whitening and wrinkle improvement effects, and thus can be usefully used as a health food.

The health food of the present invention may be added as a complex fermented extract of bokbunja, soybean, and herb, or may be used together with other foods or food ingredients, and may be suitably used according to a conventional method.

There are no particular restrictions on the type of food. Examples of foods to which the complex fermented soybean, soybean, and wild pepper complex fermentation extracts can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, and various soups , Beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and includes all healthy foods in the usual sense.

The healthy beverage composition of the present invention may contain various flavoring agents or natural carbohydrates, etc., as additional components, like a conventional beverage. The aforementioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g per 100 ml of the composition of the present invention, preferably about 0.02 to 0.03 g.

In addition to the above, the complex fermented extract of the present invention, soybean and pepper, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers , Preservatives, glycerin, alcohol, carbonic acid used in carbonated beverages, and the like.

In addition, the complex fermented extract of the compound of the present invention, soybean and pepper, may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients can be used independently or in combination. The proportion of these additives is not critical, but is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The composition comprising a complex fermented extract of bokbunja, soybean, and green pepper prepared according to the present invention has excellent antioxidant, anti-aging, anti-inflammatory, skin whitening and wrinkle improvement effects, and can be used as a cosmetic composition, pharmaceutical composition, and health food .

Figure 1 shows the manufacturing process of the complex fermentation extract of bokbunja, soybean and sancho.
Figure 2 relates to the phylogenetic strain used in the present invention Pediococcus pentosaceus ( Pediococcus pentosaceus ) B1 is a schematic diagram.
Figure 3 relates to the results of measuring the antioxidant activity before and after fermentation in the process of preparing a complex fermentation extract of bokbunja, soybean and sancho according to the present invention.
Figure 4 relates to the results of measuring the cytotoxicity of the complex fermentation extract of bokbunja, soybean and wild pepper according to the present invention.
Figure 5 relates to the results of measuring the NO (Nitric Oxide) inhibitory activity of the complex fermentation extract of bokbunja, soybean and wild pepper according to the present invention.
Figure 6 relates to the results of measuring the anti-inflammatory inhibitory activity of the complex fermentation extract of bokbunja, soybean and wild pepper according to the present invention.
Figure 7 relates to the results of measuring the tyrosinase inhibitory activity of the complex fermentation extract of bokbunja, soybean and wild pepper according to the present invention.
Figure 8 relates to the results of measuring the melanin production inhibitory activity of the complex fermentation extract of bokbunja, soybean and sancho according to the present invention.
Figure 9 relates to the results of measuring the collagen production activity of the complex fermentation extract of bokbunja, soybean and wild pepper according to the present invention.

Hereinafter, the present invention will be described in more detail through examples and experimental examples, but they do not limit the scope of the present invention.

Example 1. Preparation of complex fermentation extract

1.1 Preparation of complex extracts

After harvesting and washing the bokbunja fruit and soybean, lyophilized and crushed to 150 mesh or more. The crushed dry bokbunja powder. Sieve soybean powder and peppercorns were sifted with a 200 mesh to uniformly unify the particles of the powder, and then mixed in a 1: 1 weight ratio, distilled water was added, and the pH was adjusted to 6.5.

The pH-controlled composite mixture was sterilized at 121° C. for 15 minutes to prepare a composite extract sample of bokbunja, soybean, and green pepper (see FIG. 1 ).

1.2 Preparation of fermented strains

Lactic acid bacteria Pediococcus pentosaceus B1 (see FIG. 2 and deposit) was inoculated in MRS medium and cultured at 37° C. for 24 hours. The cultured strain was centrifuged at 3,000 rpm, 4 ^o C for 15 minutes to recover the cells, and the cells were washed three times with a phosphate buffer solution. For the washed cells, a strain solution of OD (Optical Density) 1.0 was prepared using a phosphate buffer (see FIG. 1).

1.3 Preparation of complex fermented extracts

The strain solution according to Example 1.2 was inoculated at a ratio of 5% by weight based on the entire sample of the composite extract according to Example 1.1.

After inoculation, fermentation was prepared by fermenting at 37°C for 5 days. The fermented product was extracted by performing extraction for 5 hours using 20% ethanol as an extraction solvent. After completely removing the solids in the extract, the complex fermentation extract was completed through freeze drying (see FIG. 1 ).

Example 2. Cytotoxicity measurement

In order to evaluate the toxicity to the cells used in the experiment, the cell viability was evaluated according to the MTT assay. Passage-cultured Raw 264.7 cells were dispensed 1×10 ⁵ cells/well in 96-well plates using DMEM medium, and cultured for 24 hours in a 37°C, 5% CO ₂ incubator. After incubation, the extracellular medium was removed, and then 100 µl of the DMEM solution composed of extracts by concentration was treated and reacted at 37° C. in a 5% CO ₂ incubator for 24 hours. As a control, an experiment was conducted in the same way by treating a DMEM solution that was not treated with a sample. After the reaction was completed, the residual medium was removed and 50 µl/well of 5 mg/ml MTT (3-(4,5-dimethylthi azol-2-yl) -2,5-diphenyltetrazolium bromide) solution was treated at 37°C and 4 h reacted. Thereafter, the MTT solution was removed, and the drying process was performed at 37°C for 30 minutes, and 100 μl of DMSO (dimethyl sulfoxide) solution was added to completely dissolve the formazan, and absorbance was measured at a wavelength of 550 nm using a microplate reader. As a control, the absorbance of cells not treated with a sample was analyzed and used, and the absorbance of the treatment group was calculated as a percentage based on 100% of the control, and the cell viability was calculated.

density
(Μg/ml) Cell viability (%) Complex extract
(complex extract) Fermentation complex extract
(Fermented complex extract) 50 98.63±1.12 109.00±2.08 100 94.39±2.81 107.90±1.35 200 86.75±2.03 102.55±2.03 300 88.97±2.56 100.47±1.26 400 86.91±2.97 98.48±1.38 500 86.80±0.88 96.26±1.14 600 62.99±1.76 81.30±1.64

As a result of measuring cell viability, it was confirmed that the complex fermentation extract had lower cytotoxicity than the complex extract without fermentation (see Table 1 and FIG. 3).

Example 3. Measurement of antioxidant activity

3.1 DPPH scavenging activity

Free radicals are known to cause aging, especially skin aging. To measure the free radical scavenging activity, a 0.2 mM DPPH (2,2-Diphenyl-1-picryl-hydrazyl) solution was prepared using methanol and then mixed with the extracts prepared for each concentration. After the mixture was reacted at 37°C for 30 minutes, absorbance was analyzed at a wavelength of 517 nm using a UV-Vis spectrophotometer. As a control, a DPPH solution without an extract was used in the formula for calculating the result value by analyzing the absorbance under the same conditions, and was measured by the following formula to confirm the DPPH radical scavenging activity using the absorbance value. .

DPPH radical scavenging activity (%) = [(Abs _DPPH -Abs _Sample )/Abs _DPPH ] × 100

Here, Abssample is the absorbance of the sample reaction solution, Absblank is the absorbance of only the sample, and Abscontrol is the absorbance of the DPPH solution.

As a result of measuring the DPPH removal activity of the complex extract prepared without the fermentation process and the complex fermentation extract produced by the fermentation process, it was confirmed that the DPPH removal activity is higher when using the complex fermentation extract prepared by the fermentation process ( See Figure 4).

2.2 Measurement of polyphenol content

Measurement of total polyphenol content (total polyphenol content) was quantified using the principle that Folin-reagent is reduced by a phenolic compound to develop a molybdenum blue color. The standard was prepared using gallic acid (Sigma) at a concentration of 0-100 µg/mL, and the polyphenol content of the sample was calculated based on the standard calibration curve obtained by analyzing in the same manner as the sample. After extracting the extract of each sample in 60% ethanol, the diluted sample and Folin-Ciocalteau's phenol reagent reagent were mixed and reacted for 1 hour while being shaded at room temperature. After the reaction, 200 μL was dispensed into 96 well plates, and the absorbance was measured at 765 nm, and the total polyphenol content eluted from 1 g of the dried sample was measured based on gallic acid.

As a result of comparing the total polyphenol content of the complex extract prepared without the fermentation process and the complex fermentation extract produced by the fermentation process, it is confirmed that the total polyphenol content is higher when using the complex fermentation extract produced by the fermentation process (See FIG. 4).

Example 4. Measurement of anti-inflammatory activity

3.1 Measurement of NO (Nitric Oxide) inhibitory activity

For the complex fermentation extract according to the present invention, the effect of alleviating the inflammatory response, which is one of the main causes of wrinkle formation, was evaluated by measuring the effect of inhibiting NO (Nitric Oxide) production.

In order to measure the effect of inhibiting NO production, RAW264.7 cells (mouse monocyte/macrophage-like cells) were added to phenol red free DMEM medium containing 10% FBS at 5×10 ⁵ cells/well in 24-well plates. I was busy. After processing the sample and taking the culture after 48 hours, the sample was treated 1:1 with Griess reagent, and then reacted for 15 minutes at room temperature, and then measured at a wavelength of 540 nm.

density
(Μg/ml) NO production inhibitory activity (%) Complex extract
(complex extract) Complex fermentation extract
(Fermented complex extract) 100 57.46±3.98 51.57±1.24 200 43.60±0.89 38.00±0.39 300 39.36±1.18 32.36±1.54 400 27.77±1.62 32.59±1.52 500 28.13±1.91 28.70±0.28

As a result of measuring the NO production inhibitory activity of the complex extract prepared without the fermentation process and the complex fermentation extract produced by the fermentation process, the NO production inhibitory activity is somewhat high when a large amount of the complex fermentation extract produced by the fermentation process is used. It was confirmed (see Table 2 and Figure 5).

3.2 Cytokine (TNF-α) inhibitory activity measurement

In the complex fermentation extract according to the present invention, in order to evaluate whether there is an effect of alleviating the inflammatory response, which is one of the main causes of wrinkles, the concentration of the inflammatory reaction mediator TNF-α is measured to confirm the inhibitory activity during the treatment of the complex fermentation extract Did.

Raw 264.7 cells were dispensed at 5×10 ⁵ cells/well in 24-well plates using DMEM medium with 10% FBS and 1% antibiotics, and cultured for 24 hours in a 37°C, 5% CO ₂ incubator. After incubation, the medium was removed and 500 μl of the extract solution according to the concentration composed of 1 ppm of LPS for TNF-α induction was treated and reacted for 24 h. Then, the culture was taken and the concentration of TNF-α was measured using a TNF-α ELISA kit.

density
(Μg/ml) TNF-α secretion inhibitory activity
(TNF-α secretion inhibitory activity, %) Complex extract
(complex extract) Fermentation complex extract
(Fermented complex extract) 100 22.81 45.03 200 41.87 51.87 300 54.72 63.28 400 65.53 67.94 500 76.88 78.31

As a result of measuring the inhibitory activity of TNF-α production of the complex extract prepared without the fermentation process and the complex fermentation extract produced by the fermentation process, the TNF-α production inhibitory activity was observed when using the complex fermentation extract prepared by the fermentation process. It was confirmed that it was higher (see Table 3 and Figure 6).

Example 4. Measurement of whitening activity

4.1 Measurement of tyrosinase inhibitory activity

To evaluate the whitening activity of the complex fermentation extract according to the present invention, tyrosinase inhibitory activity was measured. Tyrosinase acts on a mechanism for synthesizing melanin and is one of the enzyme factors that induce melanin formation.

The tyrosinase inhibitory activity of the sample was measured using B16F10 melanoma. B16F10 melanoma was dispensed in 2 mL by using each cell medium containing 10% FBS in a 6 well plate to be 1×10 ⁴ cells per well, and cultured for 24 hours in a 37°C, 5% CO ₂ incubator. After confirming the adhesion of cells, the samples were exchanged for each concentration, and after 1 hour, after 100 nM α-MSH stimulator treatment, the cells were incubated for 2 days at 37°C and 5% CO ₂ incubator. After removing the medium, the Lysis buffer to which Triton X-100 and 100 mM PMSF was added was treated and centrifuged at 15,000 rpm for 10 minutes. Thereafter, the mixture buffer mixed with 2% N,N-dimethyl formamide and 5 mM L-DOPA was mixed with the centrifuged cell supernatant to react and absorbance was measured at 475 nm. The inhibitory activity of tyrosinase was expressed as a percentage compared to the group treated with α-MSH only.

density
(Μg/ml) Tyrosinase inhibitory activity
(tyrosinase inhibitory activity, %) Complex extract
(complex extract) Fermentation complex extract
(Fermented complex extract) 100 22.19±2.98 23.18±3.53 200 29.37±2.08 38.86±1.59 300 43.27±1.82 45.70±2.69 400 45.19±1.63 50.56±0.89

As a result of measuring the tyrosinase inhibitory activity to measure the whitening activity of the complex fermentation extract, it was confirmed that the tyrosinase inhibitory activity increased according to the amount of use. In addition, in the same amount, it was found that the effect of the complex fermentation extract prepared by the fermentation process is higher than that of the complex extract prepared without the fermentation process (see Table 4 and FIG. 7).

4.2 Measurement of melanin production inhibitory activity

In order to evaluate the whitening activity of the complex fermentation extract according to the present invention, melanin production inhibition inhibitory activity was measured.

In order to examine the effect on the biosynthesis of melanin in the sample, B16F10 melanoma was dispensed to 1×10 ⁴ in a 6 well plate and cultured for 24 hours in a 37°C, 5% CO ₂ incubator. After confirming the adhesion of the cells, in order to promote melanin production, 10% FBS was added to the medium containing each concentration and cultured for 4 hours, and then treated with 100 nM α-MSH stimulant and cultured for 2 days. The culture was removed, washed with PBS and scrapped. After centrifugation at 15,000 rpm for 20 minutes, the supernatant was removed and 1 N NaOH solution containing 10% DMSO was added to the pellet to dissolve at 80°C for 1 hour, and absorbance was measured at 475 nm using a microplate reader. Did. 100 nM α-MSH was used as a stimulant to find out about over-produced melanin in melanoma cells for external stimulation. The experiment was repeated three or more times, and the graph was compared with the sample-free group.

density
(Μg/ml) Melanin content (Melanin Content, %) Complex extract
(complex extract) Complex fermentation extract
(Fermented complex extract) 100 98.04±0.77 94.56±1.04 200 90.49±1.8 79.03±0.80 300 76.77±1.45 74.78±0.94 400 61.52±0.95 59.94±0.80

As a result of measuring the melanin synthesis inhibitory activity to measure the whitening activity of the complex fermentation extract, it was confirmed that the melanin synthesis inhibitory activity increases according to the amount of the complex fermentation extract. In addition, in the same amount, it was found that the effect of the complex fermentation extract prepared by the fermentation process is higher than that of the complex extract prepared without the fermentation process (FIG. 9).

Example 5. Measurement of wrinkle improvement activity

In order to evaluate the wrinkle improvement activity of the complex fermentation extract according to the present invention, collagen production activity was measured.

Using human fibroblast cells, the effect on the collagen production of the sample was confirmed. Fibroblast cells were dispensed 1×10 ⁴ per well into a 96-well plate using Dermal fibroblast growth medium medium, and cultured for 24 hours in a 37°C, 5% CO ₂ incubator to attach the cells. Subsequently, the medium solution prepared for each extract concentration was treated, and after 24 h, the concentration of collagen generated in the cell culture was measured using a Procollagen Type I C-peptide EIA kit.

It was found that the effect of the complex fermentation extract prepared by the fermentation process was higher than that of the complex extract prepared without the fermentation process (FIG. 9 ).

Example 6. Preparation of cosmetic composition

A cosmetic composition was prepared under the conditions shown in Table 6 (unit, weight percent).

Raw material name Content (% by weight) Purified water Balance Glycerin 5.000 Sodium hyaluronate 0.010 hydrogel PFKC 1.500 Dissolvine NA2-S 0.020 Betaine 1.500 carbopol 980 0.250 1.3ㆍBG 1.000 Keltrol F 0.060 CS-165 1.200 ARLACEL 60 0.800 TWEEN 60 0.800 M.C.T 5.000 Lanetto O 1.500 Lipex shea soft 0.100 Tocopheryl acetate 0.100 Bees wax 0.500 Dimethicone 6CS 0.300 TEA 0.250 SEPPIPLUS 400 0.150 Spices a very small amount Complex fermented extract of Bokbunja, Soybean and Sancho 7.000 Phenoxyethanol 0.300 1,2-HEXANEDIOL 0.400

Example 7. Preparation of the drug containing the extract

The pharmaceutical composition was prepared under the conditions (unit, weight%) shown in Table 7 below.

Composition Content (% by weight) Complex fermented extract of Bokbunja, Soybean and Sancho 2 saccharin 0.8 Party 25.4 glycerin 8 Spices 0.04 ethanol 4 Sorbic acid 0.4 Distilled water 59.36

Example 8. Preparation of healthy food containing extracts

25 parts by weight of the complex fermented extract of bokbunja, soybean and sancho were added to the pastry dough, and bread, cake, cookies, crackers and noodles were prepared using this mixture (see Table 8).

Composition Content (parts by weight) Pastry dough flour 100 parts by weight milk 10 to 20 parts by weight egg 10 to 20 parts by weight Sugar 5 to 10 parts by weight Baking powder 0.5 to 1.5 parts by weight Baking soda 0.5 to 1 part by weight butter 5 to 10 parts by weight Complex fermentation extract of Bokbunja, Soybean, and Sancho 25 parts by weight compared to 100 parts by weight of the pastry dough

Simple modifications or changes of the present invention can be easily carried out by those skilled in the art, and all such modifications or changes can be considered to be included in the scope of the present invention.

Depository name: Institute for Agricultural Biotechnology

Accession number: KACC81067

Date of Deposit: 20171124

Claims (10)

A cosmetic composition containing a complex fermented extract of bokbunja, soybean and sancho as an active ingredient.
According to claim 1, wherein the complex extract of bokbunja, soybean and sancho, characterized in that the bokbunja, soybean and sancho are mixed in a weight ratio of 1: 1, cosmetic composition.
According to claim 1,
The active ingredient is produced by fermenting a complex extract of bokbunja, soybean, and sancho using lactic acid bacteria, cosmetic composition.
According to claim 3,
The lactic acid bacteria is Pediococcus pentosaceus B1 (KACC 81067BP), cosmetic composition.

According to claim 3,
The fermentation is characterized in that carried out for 2 to 7 days at 37 ℃, cosmetic composition.
According to claim 1,
The cosmetic composition is characterized in that the antioxidant, anti-aging, anti-inflammatory, skin whitening and wrinkle improvement purposes, cosmetic composition.
According to claim 1,
The active ingredient is characterized in that it contains 100 to 500㎍ / mL, cosmetic composition.
According to claim 1,
The cosmetic composition is a cosmetic composition having any one formulation selected from the group consisting of lotion, essence, lotion, cream, pack, foundation, gel, ointment and spray.
Antioxidant, anti-aging and anti-inflammatory pharmaceutical composition containing a complex fermented extract of bokbunja, soybean and sancho as an active ingredient.
A health food for improving antioxidant, anti-aging, anti-inflammatory, skin whitening and wrinkles, which contains a complex fermented extract of bokbunja, soybean, and sancho as an active ingredient.

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