KR101775485B1 - Cosmetic composition containing fermentative extract of Spatholobus suberectus Dunn as active ingredient - Google Patents

Abstract

The present invention relates to a blood- suberectus Dunn ) fermented extract as an active ingredient, characterized in that it contains 0.0001 to 30.0% by weight, based on the total weight of the cosmetic composition, of a fermented extract such as a blood-brain extract as an effective ingredient. According to the present invention, the fermentation extract of the blood-blood-sucking fermentation extract has an effect of inhibiting MMP-1 production, collagen biosynthesis effect, melanin production inhibitory effect, antioxidative effect, skin wrinkle improving effect, skin elasticity improvement, moisturizing effect, Can be provided.

Description

[0001] The present invention relates to a cosmetic composition containing a fermented extract such as blood, blood, or the like as an active ingredient.

The present invention relates to a cosmetic composition containing an extract of fermented blood as an active ingredient.

A variety of physical and chemical changes occur in human skin during the aging process. The cause of this phenomenon is divided into intrinsic aging and photo-aging. Free radicals can be caused by activation of ultraviolet rays, stress, disease state, environmental factors, wounds, and aging. When such state is deepened, the antioxidant defense network existing in the living body is destroyed, And aging.

More specifically, lipids, proteins, polysaccharides and nucleic acids, which are major components of the skin, are oxidized to destroy skin cells and tissues, resulting in skin aging. In particular, the oxidation of proteins can cause collagen (collagen), hyaluronic acid, elastin, proteoglycan, fibronectin, and the like that are severely hyperinflammatory reactions and skin elasticity If this gets worse, DNA mutations can lead to mutations, cancer, and immune deficiency.

Therefore, it is necessary to protect the cell membrane by destroying the free radicals mediated by free radicals, ultraviolet rays, and inflammation that occur during the metabolism of the body, and to regenerate already damaged cells by active metabolism, Can quickly recover and maintain healthy skin.

Aging involves not only these free radicals, but also enzyme called matrix metalloproteinase (MMP), a collagenase. That is, synthesis and degradation of extracellular matrix such as collagen in vivo are appropriately controlled, but collagen synthesis is reduced as aging progresses, and expression of matrix metalloproteinase (MMP), an enzyme that degrades collagen, is promoted, And the wrinkles are formed. These degrading enzymes are also activated by ultraviolet irradiation.

Therefore, there is a demand for development of a substance capable of modulating MMP expression induced in the cell or inhibiting its activity. Until now, the raw materials used as cosmetic materials mostly inhibited only the enzyme activity. Therefore, we tried to control the amount and activity of MMP expression induced by intracellular natural aging and photoaging.

On the other hand, skin changes due to pigment deposition occur. The factors that determine skin color are basically the partial or total pigmentation such as stain, freckle and tanning due to ultraviolet exposure, and the distribution of acne and scars and keratin Blood circulation, stress, and health. The most important factor among these factors is pigmentation.

Melanin, melanoid, carotene and hemoglobin are the most important factors affecting skin color. Among them, melanin is the most important factor affecting the biosynthesis of melanin.

Melanin absorbs or scatters ultraviolet light and plays a major role in preventing damage to the skin from ultraviolet rays. It does not have a specific maximum absorption wavelength and absorbs light in all areas. In addition, it has excellent ability to remove reactive oxygen species, sometimes melanin itself generates active oxygen, and other substances are reduced or oxidized by catechol or quinone in the melanin structure, and melanin itself shows free radical properties Pray.

The biosynthesis of melanin begins with the conversion of tyrosine, an amino acid, into melanosomes of melanocyte-forming melanosomes by tyrosinase and conversion to dihydroxy phenylalanine, followed by a series of oxidation processes to form pheomelanin, eumelanin.

This biosynthesis process is carried out in a special form of melanocyte in the brown intracellular organelle. The melanomas, including melanin granules, migrate from the nucleus to the tip of the dendritic process, and migrate into the cytoplasm by phagocytosis of keratinocytes. It accumulates around the nucleus.

The synthesis of melanin, the number of melanosomes, and the migration to keratinocytes in the periphery are partially affected by hormones and ultraviolet rays, and are primarily affected by genetics. In addition, it is known that interleukins, prostaglandins, histamines, and the like are involved in the expression of tyrosinase and cytokine, which is an intracellular regulator involved in the synthesis and transmission of melanin, and metal ions such as copper, zinc and iron.

(Japanese Patent Laid-open No. 4-9320), hydroquinone (Japanese Patent Laid-Open No. 192062), kojic acid (Japanese Patent Laid-open No. 56-7710), arbutin Activity, and is used as a whitening cosmetic. However, since it is low in stability in cosmetic formulations, its use is restricted due to decomposition and coloring, generation of odor, efficacy at a living body level, unclear effect and safety problem. These substances have proved their tyrosinase inhibitory effect, but their effect is lower in experiments similar to actual living body level. Therefore, it is required to evaluate the inhibitory effect by the melanoma cell culture similar to the biological level. Hydroquinone is also defined as a carcinogenic substance and its use in cosmetics is prohibited or restricted. Kojic acid and ascorbic acid are very unstable substances. When a cosmetic containing a small amount of the above components is kept at room temperature for several weeks, browning occurs.

In order to solve these various problems, recently, many cosmetics using natural products have been developed to reduce skin irritation caused by various chemical substances and the like. Natural materials are not only less harmful to the skin, but also have increased their value as a raw material for cosmetics due to the recent popularity of cosmetics using natural materials.

Korean Patent Laid-Open Publication No. 2010-0120766 discloses a cosmetic composition for improving skin wrinkles containing blood, etc., and Korean Patent Laid-Open No. 2006-0064112 discloses an antioxidant composition which is free from radical scavenging and production inhibition It has been disclosed that it has an efficacy.

The inventors of the present invention have studied the applicability of various natural products to cosmetics as a result of studying the possibility of applying them to cosmetics. As a result, the present inventors have found that fermented extracts can be used to produce MMP-1 production inhibitory effect, antioxidative effect, collagen biosynthesis effect, Production inhibitory effect, moisturizing effect, skin wrinkle improvement effect, skin elasticity improvement effect, and skin antiaging effect, and as a result, it was found that the efficacy as a cosmetic product can be expected.

It is an object of the present invention to provide a pharmaceutical composition containing a fermented extract such as a blood-brain extract and exhibiting an effect of inhibiting MMP-1 production, collagen biosynthesis effect, melanin production inhibitory effect, antioxidative effect, moisturizing effect, skin wrinkle improving effect, skin elasticity improving effect, To provide a cosmetic composition.

Accordingly, the object of the present invention as described above is achieved by a cosmetic composition comprising an extract of fermented blood as an active ingredient.

In addition, the fermentation extract such as blood-borrowing may be fermented with at least one microorganism selected from the group consisting of yeast, lactic acid bacteria and Bacillus.

Preferably, the fermentation extract is obtained by fermentation with Saccharomyces cerevisiae ATCC 18824. [

Preferably, the lactic acid bacteria may be Lactobacillus, Bifidobacterium, Streptococcus, Leuconostock, Pediococcus, Lactococcus, and mixtures thereof.

Preferably, the Bacillus subtilis may be Bacillus subtilis, Bacillus subtilis, Bacillus licheniformis, Bacillus megaterium, Bacillus polyfermenticus, Bacillus mesentericus and mixtures thereof.

Preferably, the cosmetic composition is for improving skin wrinkles.

Further, preferably, the cosmetic composition is characterized by being for skin whitening.

Preferably, the cosmetic composition is characterized by being for antioxidation.

Preferably, the cosmetic composition is characterized in that it is for moisturizing.

Preferably, the cosmetic composition is for improving skin elasticity.

Preferably, the cosmetic composition is characterized in that it is for preventing skin aging.

Preferably, the fermented blood-extracting extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition.

Preferably, the cosmetic composition is a cosmetic composition exhibiting an effect of inhibiting MMP-1 production, collagen biosynthesis effect, melanin formation inhibitory effect, moisturizing effect, skin wrinkle improving effect, skin elasticity improving effect, .

Preferably, the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream and spray Lt; RTI ID = 0.0 > of: < / RTI >

The fermentation extract of the present invention was found to have an effect of suppressing MMP-1 production (Experimental Example 1), an effect of enhancing collagen synthesis (Experimental Example 2), an effect of inhibiting melanin production (Experimental Example 3), an antioxidative effect (Experimental Example 4) (Experimental Example 5), wrinkle improving effect (Experimental Example 6), and skin elasticity improving effect (Experimental Example 7).

(Also called blood spots, blood spots, active blood spots, red light, blood spots, etc.) used as an active ingredient in the cosmetic composition of the present invention include Spatholobus suberectus It is labeled as Dunn . It is a medicinal product made by drying the stem of wheat tofu, which is a soybean and plant of rosemary roots, Mucunae Caulis. Dicotyledonous plant phyllotaxis Rice roots Dried stem of bean flour (dense flower bean), which is used as medicinal herb and origin in China. The small stem of the wheat earthenware was cut and dried in the sun, and the juice of the stem was red like the blood of a chicken. In one room, it acts on the heart and spleen to help the blood circulation, supplement the blood, and eliminate the blood. In addition, there is a function to make menstruation even more women, menstrual irregularities, menstrual pain, amenorrhea is also used. Blood circulation and blood circulation promoting action, such as menstrual cramps, menstrual irregularities and is effective in the effects of limb paralysis. It is also used when you are weak and has anemia. It also works for hypertension, heart disease and angina. Pharmacological actions include inflammation, analgesic action, kidney and uterine metabolic activation, uterine stimulation, coronary artery dilatation, and leukocyte hyperplasia.

In the present invention, the blood-brain-like extract means the one obtained by extracting blood, etc.

The blood or the blood of the present invention may be finely pulverized by 100 to 500 mesh, such as blood or blood, which has been thoroughly washed and dried, and various organs or parts such as blood can be used.

In the present invention, the fermentation extract such as a blood-borne and the like means obtained by fermentation of an extract such as blood-brain. The extract may be prepared by using a conventional solvent under the conditions of ordinary temperature and pressure, but is preferably (a) water, an anhydrous or a lower alcohol having 1 to 4 carbon atoms (e.g., methanol, ethanol Propanol and butanol), a solvent selected from the group consisting of propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane, Solvent extraction, (b) decompression by carbon dioxide, supercritical extraction by high temperature, or (c) ultrasonic extraction.

In addition, a fermentation extract such as a blood-blooded fermentation extract obtained by fermenting microorganisms inoculated with a culture medium obtained by adding the above-mentioned blood-cell extract etc. to a normal microorganism culture medium is used.

The fermentation extract of the present invention may be prepared by adding 1 to 30% by weight of an extract of a blood culture extract to the total culture medium weight in a culture medium, inoculating the fermentation microorganism in an amount of 100,000 to 10,000,000 cfu / L, Deg.] C, preferably 25 to 35 [deg.] C for 12 to 168 hours, preferably 12 to 72 hours.

In the culture medium used for fermentation, the amount of the extract such as a blood-brain extract may be 1 to 30% by weight based on the total culture medium weight. If the extract is less than 1% by weight, the amount of the fermentation broth due to the microorganism is small and the effect is difficult to be exhibited. If the extract is more than 30% by weight, the fermentation efficiency is decreased because the extract such as blood-

The fermenting microorganism used in the present invention may be at least one selected from the group consisting of yeast, lactic acid bacteria, and bacillus, preferably Saccharomyces, more preferably, Saccharomyces cerevisiae ATCC 18824 is used.

The lactic acid bacteria that can be used in the present invention preferably include Lactobacillus, Streptococcus, Bifidobacterium, Leuconostock, Pediococcus, Lactococcus and mixtures thereof, most preferably Lactobacillus . Among lactobacilli, the most preferred lactic acid bacterium for use in fermenting the mist tree of the present invention is Lactobacillus plantarum .

Bacillus bacteria which can be used in the present invention preferably include Bacillus subtilis, Bacillus fumillis, Bacillus licheniformis, Bacillus megaterium, Bacillus polyfermenticus, Bacillus mesentericus, and mixtures thereof And most preferably Bacillus subtilis .

After fermentation, the culture broth is centrifuged to separate only the supernatant to obtain a fermentation extract such as blood. The fermented extract thus obtained is used as it is, or the fermentation broth is dried and used. When the product is dried and used, an extraction solvent of 1 to 20 times the dry weight is added and the product is immersed at 4 to 30 ° C for 3 to 20 days to extract an active ingredient. Extraction with an extraction solvent may be carried out using any extraction solvent conventionally used in the art.

The extract was filtered through Whatman # 10 filter paper, and the filtrate was finally filtered using 0.25 μm filter paper. The filtrate was concentrated under reduced pressure and freeze-dried at 60 ° C or lower to obtain a filtrate, The fermentation extract is finally obtained using an extraction solvent such that it contains 70% by weight.

According to the present invention, the blood fermentation extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition, more preferably 0.01 to 10% by weight . When the content of the extract is less than 0.0001% by weight, the effect of improving the skin is not exhibited. When the content of the extract is more than 30.0% by weight, the degree of improvement of the skin against the increase of the content is insignificant, It is not even.

Therefore, the cosmetic composition of the present invention including the blood-blooded fermentation extract having various functions has excellent collagen synthesis enhancing effect, MMP-1 production inhibiting effect, antioxidant effect, skin wrinkle improving effect, skin elasticity improving effect, skin whitening effect, Melanin production inhibitory effect, moisturizing effect, and skin aging prevention effect.

The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients commonly used in cosmetic compositions in addition to the above-mentioned effective ingredients, and may contain conventional ingredients such as antioxidants, stabilizers, solubilizers, vitamins, Supplements, and carriers.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powdered foundation, emulsion foundation, wax foundation, pack, massage cream and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide ether Alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation or a surfactant-free cleansing formulation, it may be applied to the skin and then wiped off, washed or rinsed with water. The surfactant-containing cleansing formulation may be a cleansing foam, a cleansing water, a cleansing towel and a cleansing pack. The surfactant-free cleansing formulation may be a cleansing cream, , Cleansing lotion, cleansing water and cleansing gel, but is not limited thereto.

On the other hand, the cosmetic process of the present invention refers to all cosmetic processes using the cosmetic composition of the present invention. That is, all methods known in the art using a cosmetic composition belong to the cosmetic method of the present invention.

The cosmetic composition of the present invention may be used alone or in combination, or may be used in combination with other cosmetic compositions other than the present invention. The cosmetic composition according to the present invention may be used according to a conventional method of use, and may be used in a number of times depending on the skin condition or taste of the user.

By carrying out the cosmetic method using the cosmetic composition comprising the fermentation extract of the present invention, it is possible to obtain an effect of inhibiting MMP-1 production, an effect of promoting collagen synthesis, an effect of inhibiting melanin generation, a moisturizing effect, have.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Manufacturing example  1 and 2: Bloodbath  Fermented extract preparation

After shaking 100g of shrimp blood and shade, it was refluxed 3 times for 5 hours with 70% (v / v) aqueous ethanol solution, cooled down and filtered with Whatman # 4 filter paper. The filtered extract was concentrated under reduced pressure at 50 DEG C or below and lyophilized to obtain 21.3 g of blood extract and the like.

Lactobacillus MRS broth (DIFCO, USA) was used as the culture medium for the lactic acid bacteria, and 10% by weight of the above-described blood-brain extract was added to the total microorganism culture medium weight .

Three saccharide in my process to the YPD medium Levy jiae (Saccharomyces cerevisiae ATCC 18824) were inoculated at 10,000,000 cfu and cultured at 30 ° C for 72 hours.

After culturing, the culture broth was centrifuged to remove the bacteria to obtain a fermented product such as blood, and the ethanol was added to the final 70% (v / v), followed by refluxing three times for 5 hours, And filtered through Whatman # 4 filter paper. The filtered fermented extract was filtered through a 0.25 μm filter paper, and the filtrate was concentrated under reduced pressure and freeze-dried at 60 ° C or lower to obtain 8.7 g of fermented extract of fermentation broth (Preparation Example 1).

Lactobacillus plantarum ATCC 10012 was inoculated at 10,000,000 cfu and cultured at 37 DEG C for 72 hours. The extract was added to Lactobacillus MRS medium in an amount of 10% by weight based on the total weight of the medium.

After culturing, the culture broth was centrifuged to remove the bacteria to obtain a fermented product such as blood and blood. Ethanol was added to the final 70% (V / V), refluxed three times for 5 hours, And filtered through Whatman # 4 filter paper. The filtered fermented extract was filtered through a 0.25 μm filter paper, and the filtrate was concentrated under reduced pressure and freeze-dried at 60 ° C or less to obtain 9.1 g of a fermented extract of fermentation broth (Preparation Example 2).

Comparative Manufacturing Example  One : Bloodbath  Solvent extract

As a comparative example of fermented extracts such as blood, blood and the like, extracts were prepared. The blood and blood extracts were washed and shaken 100g of shrimp blood, and then refluxed with 70% (v / v) aqueous ethanol solution for 5 hours for 3 hours. After cooling, the mixture was filtered through Whatman # 4 filter paper . The filtered extract was concentrated under reduced pressure at 50 DEG C or below and lyophilized to obtain 21.3 g of blood extract and the like.

Experimental Example  One : Bloodbath  Fermented extract MMP -1 production inhibitory effect

In Experimental Example 1, the effect of inhibiting the production of collagenase, MMP-1, from the fermentation extracts obtained in Preparation Examples 1 and 2 was measured.

Human normal skin cells were inoculated into a 48-well microplate (Nunc, Denmark) at a concentration of 1 × 10 ⁶ cells per well and cultured in DMEM medium (Sigma, USA) and 37 ° C For 24 hours, and then cultured for 48 hours in serum-free DMEM medium supplemented with Preparation Examples 1 and 2 and Comparative Preparation Example 1 and in serum-free DMEM medium containing no fermentation extract.

After the incubation, the supernatant of each well was collected and the amount (ng / ml) of the newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, USA) The inhibition rate (%) was calculated and the results are shown in Table 1.

Treatment concentration Inhibition rate of MMP-1 production (%) Production Example 1 Production Example 2 Comparative Preparation Example 1 10ppm 51.39 52.07 23.12 25 ppm 80.13 77.45 41.45 50 ppm 94.54 92.71 61.24 TGF-beta (200 nM) 79.52

As shown in the results of Table 1, it was confirmed that the fermentation extract of the present invention of the present invention inhibits MMP-1 production (inhibits MMP-1 activity) in a concentration-dependent manner. Even considering the difference between the concentration of TGF-beta (200 nM), a substance known to have an inhibitory effect on MMP-1 production, and the concentration of fermentation extract (500 ppm) of the present invention, The same degree of inhibition of MMP-1 indicates that the fermentation extract of the present invention has an inhibitory effect on MMP-1 production.

Experimental Example  2. Bloodbath  Effect of fermentation extract on collagen synthesis

Human normal epithelial cells, fibroblasts, were inoculated into 48-well microplates at a density of 1 x 10 ⁶ cells per well and cultured in DMEM medium for 24 hours. Serum-free DMEM medium supplemented with Preparation Examples 1 and 2 or Comparative Preparation Example 1 and a control group were further cultured in serum-free DMEM medium containing no fermentation extract for 48 hours. After the incubation, the supernatant of each well was collected, and the amount of procollagen type I C-peptide (PICP) was measured using a collagen kit (Takara, Japan) and converted into ng / Respectively. Collagen biosynthesis increase rate (%) was calculated according to the following formula (2), and the results are shown in Table 2.

Treatment concentration Collagen biosynthesis rate (%) Production Example 1 Production Example 2 Comparative Preparation Example 1 2.5 ppm 21.2 23.3 7.3 5 ppm 40.1 41.8 11.6 10ppm 62.6 68.4 23.9 25 ppm 125.3 120.1 68.8 50 ppm 165.9 161.5 105.1 TGF-beta (200 nM) 167.7

According to Table 2, it was confirmed that the collagen biosynthesis rate of the fermentation broth obtained by Preparation Examples 1 and 2 of the present invention was increased in a concentration-dependent manner. In particular, it can be confirmed that the effect of the fermentation extract prepared in Production Examples 1 and 2 is better than that of Comparative Production Example 1. [

Experimental Example  3: Bloodbath  Inhibitory Effect of Fermented Extracts on Melanin Production by B16F1 Melanocytes

Experimental Example 3 was performed to determine the whitening effect of B16F1 melanin-forming cells in order to examine the whitening effect of the fermentation extracts obtained from the above-mentioned Preparation Examples 1 and 2. The B16F1 melanin-forming cell used in Experimental Example 3 is a cell strain derived from a mouse, and is a cell that secretes a melanin pigment called melanin.

During the artificial culture of these cells, samples were treated to compare the degree of reduction of melanin pigment. The B16F1 melanin-forming cells used in this experiment were purchased from the American Type Culture Collection (ATCC).

Melanin biosynthesis inhibitory effect of B16F1 melanin-forming cells was measured as follows.

B16F1 melanocyte-forming cells were divided into 6-well plates at a concentration of 2 × 10 ⁵ cells per well. Cells were adhered and treated with the fermentation extracts of the fermentation broths according to Preparation Examples 1 and 2 at a concentration not causing toxicity, and cultured for 72 hours . After incubation for 72 hours, the cells were detached with trypsin-EDTA, the number of cells was measured, and the cells were recovered by centrifugation. Quantification of intracellular melanin was carried out with a slight modification of the method of Lotan (Cancer Res., 40: 3345-3350, 1980). Cell pellet was washed once with PBS, and 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt the cells. After centrifugation (3,000 rpm, 10 min), 1N NaOH (10% DMSO) was added to the cell filtrate to dissolve the extracted melanin and the absorbance of melanin was measured at 405 nm with a microplate reader. The inhibition rate (%) of melanin formation was measured. Melanin formation inhibition rate (%) of B16F1 melanin-forming cells was calculated according to the following equation (3), and the results are shown in Table 3 below.

A: Amount of melanin in wells to which no sample was added

B: Amount of melanin in the well to which the sample was added

Treatment concentration (% by weight) Melanin formation inhibitory effect (%) Production Example 1 Production Example 2 Comparative Preparation Example 1 0.001 21.37 22.75 12.43 0.005 47.85 48.42 22.11 0.01 65.43 67.82 31.32 0.05 79.56 82.21 45.42 Arbutin 200ppm 69.53

As can be seen from the results of Table 3, it can be seen that the fermentation extracts prepared by Preparation Examples 1 and 2 of the present invention significantly inhibited melanin production in B16F1 melanin-forming cells.

Experimental Example  4: Bloodbath  Fermented extract DPPH Radical Scatters  analysis

In Experimental Example 4, the antioxidative effect (free radical scavenging ratio) was measured by the DPPH method in order to measure the antioxidative effect of the fermentation extracts obtained in Preparation Examples 1 and 2. The DPPH method measures the antioxidative effect by reducing power using a free radical called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. The degree of free radical scavenging (%) at a wavelength of 560 nm is measured by comparing the degree of decrease in absorbance of DPPH by the test substance (fermentation extract such as blood) and the absorbance of the blank test solution. As a reagent used, 61.88 mg of a 0.1 mM solution of 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76) was dissolved in methanol to make 100 ml.

As a measuring method

(1) Add 0.15 ml of the sample solution to 0.15 ml of 0.1 mM DPPH solution in a 96-well plate, stir rapidly, and start culturing at 25 ° C for 10 minutes.

② Measure the absorbance St at 560 nm thereafter.

(3) The blank of the sample solution is measured by the same procedure as that for measuring the absorbance St except that methanol is used instead of the 0.1 mM DPPH solution.

(4) The absorbance Bt is measured by operating in the same manner as in the measurement of the absorbance St except that distilled water is used instead of the sample solution.

(5) Blank of the blank test is carried out in the same manner as in the measurement of the absorbance St except that methanol is used instead of the 0.1 mM DPPH solution and distilled water is used instead of the sample solution, and the absorbance Bo is measured.

The results of the free radical scavenging rate (%) were calculated by the equation (4), and the results are shown in Table 4 below.

St: absorbance at 514 nm after free radical scavenging of the sample solution

Bt: Absorbance at 514 nm after free radical scavenging of blank test solution

So: the absorbance at 514 nm before the reaction in the absence of free radicals in the sample solution

Bo: absorbance at 514 nm before the reaction in the absence of the free radical of the blank test solution

Name of sample SC ₅₀ (%) Production Example 1 0.00015 Production Example 2 0.0001 Comparative Preparation Example 1 0.001

SC ₅₀ , which is the concentration of the sample required for 50% removal of the DPPH free radical, was confirmed. As a result, Production Examples 1 and 2 showed antioxidative effects.

Formulation Example  One : Bloodbath  Fermented extract Cosmetics  Preparation of composition

As shown in the following Table 5, cosmetic preparations (Formulations 1 and 2) each containing fermented extracts of blood culture and fermentation extracts of Preparation Examples 1 and 2 were prepared, and for the comparison of efficacy, (Comparative Formulation Example 1) containing a blood-extracting extract of Comparative Preparation Example 1 instead of the fermented extract, a cosmetic composition comprising glycerin and 1,3-butylene glycol as a moisturizing agent instead of a fermentation extract such as a blood- 2), a cosmetic composition containing glycerin and sorbitol as a moisturizing agent (Comparative Formulation Example 3), and a cosmetic composition containing no extract or moisturizer (Comparative Formulation Example 4).

(Unit: wt%) division ingredient Formulation Example
One Formulation Example
2 compare
Formulation Example 1 compare
Formulation Example 2 compare
Formulation Example 3 compare
Formulation Example 4 A Fermentation extract such as blood and blood
(Production Example 1) 5.0 - - - - - Fermentation extract such as blood and blood
(Production Example 2) - 5.0 - - - - Extract of blood etc.
(Comparative Production Example 1) - - 5.0 -  -  - glycerin - - - 5.0 - - 1,3-butylene glycol - - - 5.0 - - Adenosine - - - - 3.0 - EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 to 100 B Cetostearyl alcohol 2.0 2.0 2.0 2.0 2.0 2.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 Microcrystalline 0.7 0.7 0.7 0.7 0.7 0.7 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Liquid paraffin 3.0 3.0 3.0 3.0 3.0 3.0 Trioctanoin 5.0 5.0 5.0 5.0 5.0 5.0 Polysorbate 1.2 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3.0 3.0 3.0 3.0 3.0 3.0 BHT 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

Experimental Example  5: Bloodbath  Moisturizing effect of fermented extract

The clinical moisturizing effects of the cosmetic compositions of Formulation Examples 1 and 2 were measured as follows. A and B groups were divided into five groups (A, B, C, D, E, and F) by randomly dividing 100 healthy men and women who feel skin dry through a questionnaire. (Comparative Formulation Example 1) containing Group B and Group C (Formulation Examples 1 and 2), group C (group 1), and Group D, glycerin and 1,3-butylene (Comparative Formulation Example 2), and a cosmetic composition (Comparative Formulation Example 4) containing no fermentation extract and humectant, such as blood, were applied to the E group twice a day for 8 weeks on the face for 8 weeks. The moisturizing effect was evaluated by Corneometer CM 820 (Corage + Khazaka, Germany) for every 2 weeks and after the start of the test. The experimental results are shown in Table 6 below.

division Before use After 2 weeks of use After 4 weeks of use Formulation Example 1 21.2 38.4 40.2 Formulation Example 2 22.2 39.5 51.3 Comparative Formulation Example 1 22.5 29.7 34.3 Comparative Formulation Example 2 22.3 36.3 45.8 Comparative Formulation Example 4 22.2 24.3 24.7

As shown in Table 6, it was confirmed that the moisturizing effect of the cosmetic compositions (Formulation Examples 1 and 2) containing the fermented extract of the present invention of the present invention was excellent in comparison with the cosmetic compositions containing other moisturizing agents (Comparative Formulation Example 2) have.

Experimental Example  6. Bloodbath  Effect of fermentation extract on skin wrinkles

Clinical trials on the effect of improving the skin wrinkles of cosmetic compositions containing the fermentation extract of the present invention of the present invention were carried out. Clinical trials were conducted through actual use tests of each cosmetic product.

In other words, the clinical tests were conducted in the same manner as in Formulation Examples 1 and 2 containing the fermentation extracts of Preparation Examples 1 and 2 of Table 5 and comparative Formulation Example 1 containing the extracts of the fermentation extracts such as blood and blood, The cosmetic composition of Comparative Formulation Example 2 containing glycerin and 1,3-butylene glycol as a moisturizing agent instead of the extract, Comparative Formulation Example 3 containing adenosine 3% (v / v) instead of a fermentation extract such as blood, And the effect of improving the skin wrinkles due to the use of the cosmetic material was confirmed and compared.

100 women (30 to 50 years old) were divided into 20 experimental groups, each containing a glycerin and 1,3-butylene glycol-containing formulation (Comparative Formulation Example 2) as a moisturizing agent instead of a fermentation extract such as a blood- (Comparative Formulation Example 3) in which the fermented extract was replaced with adenosine 3% (v / v), the experimental group of the formulation (Formulation Example 1) containing Preparation Example 1, the formulation containing Preparation Example 2 ) And the experimental group of the formulation containing Comparative Preparation Example 1 (Comparative Formulation Example 1), and the wrinkle improvement effect after 6 weeks was visually evaluated using both sides of the face to check the wrinkle improvement degree. The results of the wrinkle-improving effect based on this evaluation are shown in Table 7 below.

The effect of improving wrinkles after the completion of the experiment was evaluated by visual examination and instrument measurement (cutometer SEM 575, C + K Electronic Co., Germany) of a skilled physician before and after 3 months of use. The experimental results are shown in Table 7 below.

Wrinkle improvement effect (visual evaluation and device evaluation) of a cosmetic composition containing a fermentation extract such as a blood- Wrinkle improvement effect Effective rate (%) Great slightly none Formulation Example 1 9 8 3 85.0 Formulation Example 2 10 8 2 90.0 Comparative Formulation Example 1 3 7 10 50.0 Comparative Formulation Example 2
(Moisturizer) 11 7 2 30.0 Comparative Formulation Example 3
(Adenosine) (12) (5) (3) (85.0)

As can be seen from the results in Table 7, the cosmetic compositions of Formulation Examples 1 and 2 containing the fermentation extract of the present invention showed a wrinkle improvement effect about 3.0 times higher than Comparative Formulation Example 2 containing moisturizing agent.

In addition, similar results were obtained with the cosmetic preparation of Comparative Formulation Example 3 containing adenosine, which is known to have a wrinkle-reducing effect instead of the fermentation extract such as blood or blood, and the fermentation extracts such as blood-sparing fermentation showed excellent wrinkle-reducing effects.

Experimental Example  7: Measurement of skin elasticity improvement effect

In order to examine the skin elasticity improvement effect of the cosmetic composition according to the formulation examples and comparative formulations containing the fermentation extract obtained from Preparation Examples 1 and 2 obtained in Production Example 1 or 2, Were measured. After dividing into four groups of 10, the first group had Formulation Example 1, the second group had Formulation Example 2, the third group had Comparative Formulation Example 1, and the fourth group had Comparative Formulation Example 2 in the morning and evening 2 times for 12 weeks, and skin elasticity was measured after 12 weeks. The elasticity of the facial region was measured using a cutometer SEM575 (MPA 580, Courage + Khazaka GmbH, Germany), and the R2 (biological elasticity) indicating skin elasticity was measured before and after application . The results are shown in Table 8 below, and the results are the average of the degree of improvement for each group.

Experimental product Skin elasticity improvement (%) Formulation Example 1 26.4 Formulation Example 2 29.6 Comparative Formulation Example 1 11.4 Comparative Formulation Example 2 6.5

As shown in Table 8, Formulation Examples 1 and 2 containing the fermentation extract of blood-blood-sucking improved skin elasticity significantly compared to Comparative Formulation Examples 1 and 2.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (10)

A cosmetic composition for improving skin wrinkles containing fermented extract such as blood or blood as an effective ingredient. A cosmetic composition for skin whitening comprising an extract of fermentation such as blood, blood as an active ingredient. A cosmetic composition for moisturizing, which comprises fermented extract such as blood, blood or the like as an active ingredient. A cosmetic composition for improving the elasticity of skin comprising an extract of fermented extract such as blood and blood as an active ingredient. A cosmetic composition for preventing skin aging comprising an extract of fermented extract such as blood or blood as an active ingredient. The cosmetic composition according to any one of claims 1 to 5, wherein the fermentation extract is fermented with at least one microorganism selected from the group consisting of yeast, lactic acid bacteria, and bacillus. 6. The cosmetic composition according to any one of claims 1 to 5, wherein the fermentation extract is obtained by fermenting Saccharomyces cerevisiae ATCC 18824. 6. The cosmetic composition according to any one of claims 1 to 5, wherein the fermentation extract is obtained by fermentation with Lactobacillus plantarum ATCC 10012. The cosmetic composition according to any one of claims 1 to 5, wherein the fermented blood extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. delete