KR101645476B1 - Cosmetic Composition containing Fermentative Extract of Hippophae rhamnoides with Increased Amount of Vitamin C Fermented by Aureobasidium pullulans - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a fermented extract of wild-caucasian black yeast. The fermented extract of Saccharomyces cerevisiae of the present invention can be effectively used for antioxidant, anti-inflammation, promotion of collagen synthesis, improvement of skin wrinkles, moisturizing, whitening, and skin barrier improvement.

Description

[0001] The present invention relates to a cosmetic composition containing a fermented extract of a bitterous tree having increased vitamin C content through black yeast fermentation,

More particularly, the present invention relates to a cosmetic composition containing a fermented extract having an increased content of vitamin C by fermenting the extract of Sanjay tree with black yeast mycelium .

Skin aging can be caused by active oxygen species activated by ultraviolet rays, stress, disease states, environmental factors, wounds, and age. When such conditions are intensified, the antioxidant defense network in the living body is destroyed, and cells and tissues are damaged, thereby promoting skin aging. Specifically, lipids, proteins, polysaccharides and nucleic acids, which are major constituents of the skin, are oxidized to destroy skin cells and tissues, resulting in skin aging. Oxidation of protein breaks down collagen, hyaluronic acid, elastin, proteoglycans, fibronectin, etc. which are skin connective tissues, causing severe inflammation reaction and affect skin elasticity. When it gets worse, mutation of DNA leads to mutation, cancer, and immune function.

Therefore, it is necessary to protect the cell membrane by eradicating reactive oxygen species that are mediated by the body's metabolic process, ultraviolet radiation, inflammatory reaction, and regenerate the already damaged cells by active metabolism to proliferate the cells. It can restore and maintain healthy skin.

On the other hand, the skin is an organ that functions as a protective film for protecting the living body from the external environment, and serves to prevent loss of biocomponents in the human body, that is, water and electrolytes, and to prevent harmful substances from entering the human body from the outside. The skin is divided into the skin layer, the dermal layer and the subcutaneous fat layer. The epidermal layer consists of keratinocytes and melanocytes. Since the keratinocytes in the outermost layer of the skin are in direct contact with the external environment, they are required to have strong resistance against physical, chemical, or material permeation and prevent moisture from being lost to the outside At the same time as zooming, it should maintain its flexibility by holding proper water on its own.

Generally, the moisture content of the skin is about 70% in the dermal layer, but decreases to the skin layer, which is about 10 to 30% in the keratin layer. The water supplied from the dermis layer mainly migrates to the upper part of the stratum corneum by passive diffusion and eventually to the outside. This is called Trans Epidermal Water Loss (TEWL). It is known that the lipid component of the keratinocyte layer and epidermal lipid, which maintains the transdermal water loss at an appropriate level, is the lipid component.

It is known that the keratinocyte layer has a hydrophilic water retention ability called "Natural Moisturizing Factor (NMF)" and plays an important role in moisturizing the skin. Normal keratinocyte layer maintains skin smoothness and softness and maintains normal body protection function when moisture is maintained at about 10 to 30%. However, when the moisture content of the keratinocyte layer is less than 10%, the skin becomes rough and the protective function of the body is lost and the aging phenomenon occurs. For example, in the case of dry skin, the keratinocyte aggregation is weakened, and a scaling phenomenon appears in which the keratinocytes are peeled off like scales from the skin surface. The reason why the skin is dried in this way is that the moisture content of keratinocyte layer is smaller than that of normal skin. In addition to this, even healthy skin may be in a harsh environment, such as wind, cold weather, sunshine, cleansing, shaving, etc., which causes water shortage, which can lead to poor skin condition.

Therefore, it is very important to properly maintain the moisture content of the keratinocyte layer. To this end, cosmetics were added with ingredients similar to sebum, moisturizers such as NMF or polyol, and the like. For example, glycerin or sorbitol having three or more hydroxyl groups as a water-soluble polyol exhibits excellent moisture resistance, but it is very sticky and feels uncomfortable when used. Propylene glycol 1,3-butylene glycol having two hydroxyl groups, . In addition, other natural moisturizing factors such as sodium pyrrolidonecarboxylate (PCA-Na), sodium lactate, urea, etc., are highly electrolytic, which deteriorates emulsion stability of the cosmetic. Amino acids, collagen, elastin, etc. are also capable of moisturizing, but also had a limited ability to moisturize.

The ability of the horny layer to retain moisture can be controlled by natural moisturizing factor (NMF), which consists of sebum components, amino acids, lactic acid, urea, and inorganic salts. And the development of a substance with high moisturizing effect is one of the important research subjects in cosmetics.

Meanwhile, in recent years, many cosmetic products using natural products have been developed to reduce skin irritation caused by various chemical substances and the like. In addition to low adverse effects on skin, natural materials have recently become increasingly appreciated as a cosmetic raw material as consumers' response to cosmetics using natural materials has increased.

As a result of studying the applicability of various natural products to cosmetics, the present inventors have found that antioxidant, anti-inflammation, promotion of collagen synthesis, improvement of skin wrinkles , Moisturizing, whitening, and skin barrier function, and completed the present invention.

Korean Patent Registration No. 1229748 discloses a cosmetic composition containing a vitamin-wood fermentation extract, which describes a cosmetic composition containing an extract of Fermented Sanjota tree as an active ingredient.

The present inventors intend to provide a natural extract which is applicable to skin cosmetics and which is harmless to human body and is highly safe, and a cosmetic composition containing it as an active ingredient.

The present invention also provides a cosmetic composition having a natural extract obtained by fermenting an extract of Liliaceae with a black yeast mycelium and an effective ingredient thereof and having antioxidant, anti-inflammatory, promoting collagen synthesis, improving skin wrinkles, moisturizing and skin barrier function The purpose.

In order to attain the above object, the present invention provides a cosmetic composition comprising a fermented extract of Sanjay wood black yeast.

Preferably, the cosmetic composition may be used for antioxidation, anti-inflammation, skin wrinkle improvement, skin whitening, skin moisturizing or skin barrier function improvement.

Preferably, the above-described fermented extract of Sanjayaku black yeast can be obtained by fermenting the extract of Sanjayaki with black yeast for 1 to 7 days while maintaining the pH at 5 to 7 under a temperature condition of 20 to 40 ° C.

In addition, the cosmetic composition may contain 0.0001 to 30.0% by weight of a fermented extract of Sanjia wood black yeast, based on the total weight of the cosmetic composition.

Preferably the cosmetic composition is in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray And can have any one of the formulations selected.

INDUSTRIAL APPLICABILITY The present invention can provide a natural extract which is applicable to skin cosmetics and which is harmless to the human body and has excellent safety, and a cosmetic composition containing it as an active ingredient.

The present invention also provides a cosmetic composition having a natural extract obtained by fermenting the extract of Liliaceae with the mycelia of black yeast and an effective ingredient thereof and having antioxidant, anti-inflammation, promotion of collagen synthesis, improvement of skin wrinkles, moisturizing, whitening and skin barrier function can do.

In addition, the present invention can provide a cosmetic composition having an increased content of vitamin C by fermenting the extract with the black yeast.

FIG. 1 shows HPLC graphs of (a) before fermentation and (b) after fermentation.

The present inventors prepared extracts from various natural materials as various raw materials by various extraction methods and conducted studies to select substances having excellent effects such as antioxidant, anti-inflammation, promotion of collagen synthesis, improvement of skin wrinkles, moisturizing and skin barrier function Respectively. As a result, it has been found that the extract prepared by fermenting with the mycelia of black yeast into the extract of Sanjay wood is most suitable for the purpose of the present invention described above.

Sanjasu ( Hippophae rhamnoides ) is a shrub that grows naturally in bad conditions such as dry breeze barren with extreme temperature difference. It is called as vitamin tree in the name of the plant. It contains 100 kinds of active substances beneficial to the human body and is excellent in antioxidant anti-inflammatory effect. Especially in the case of the fruit tree, vitamin content is the highest among the fruits, 200 to 800 times of the apple is known as the report of vitamins. In addition, it grows about 1 ~ 2m per year with the number of properties. It is strong against pests, has excellent wild planting ability, has excellent self-fertilization ability and can be cultivated without pesticide, and whole berries and leaf root trees suitable for the well-being period can be used as food tea, medicines or cosmetic raw materials.

Aureobasidium pullulans is a microorganism used to make beer and wine, a fungus or mushroom herd, but is a generic name for single celled organisms that do not have mycelium, photosynthetic activity, or motility. It is a microorganism that humans have used for food since 5000 years ago, and yeast itself is used as a cheap fat or protein source. It is rich in vitamin B group and contains vitamin D. In addition, black yeast is a microorganism that produces beta-glucan, and it is known that the black yeast fermented product alone exhibits a moisturizing effect, an antioxidant, and soothing effect. Black yeast beta-glucan contains a large amount of components known to have physiological activities such as anticancer and insulin secretion inhibition.

Until now, the extract of Sanjay has been widely used for other purposes. However, as in the present invention, the effect of the fermentation form, particularly the black yeast fermentation extract of Sanjayura, has not been specifically known.

The cosmetic composition of the present invention comprises a substance obtained by fermenting a fermented extract of Sanjay wood with a plant extract of Sanjay tree with a mycelium of Aureobasidium pullulans and exhibiting excellent antioxidation, anti-inflammation, promotion of collagen synthesis, improvement of skin wrinkles, moisturizing, whitening, It shows the function improvement effect.

In the present invention, the extract of Liliaceae can be prepared according to a conventional method known in the art. Specifically, the fruit, leaf or stem of the spruce tree can be dried and crushed, and then, using a conventional solvent under the conditions of ordinary temperature and pressure. Specifically, the extract is extracted using a solvent selected from the group consisting of water, an anhydrous or lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol, Leaves, or stems of the above-mentioned cells by using the above-mentioned solvent, and then mixing them.

The solvent is preferably an alcohol, more preferably ethanol. At this time, the ethanol is preferably 70% ethanol. The suitable amount of the extraction solvent is 1 to 15 times, more preferably 5 to 10 times, and most preferably 7 times the weight of the dried potato tree.

On the other hand, the acid extract of the present invention includes not only the extract obtained by the above-mentioned solvent extraction method but also the extract obtained through a conventional purification process. For example, by separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatographies (made for separation by size, charge, hydrophobicity or affinity), and the like, It is interpreted that the fraction is also included in the extract of the present invention.

In addition, the plant extract of the present invention may be prepared in powder form by an additional process such as vacuum distillation, freeze drying, spray drying and the like.

The extract of the wild-caught black yeast black yeast can be prepared by fermenting the above-mentioned extract of Sanjay tree with black yeast. Specifically, it can be produced by the following method. First, 1 to 5% by weight of glucose is added as a carbon source to the extract of Sanjay tree, and 0.1 to 1% by weight, preferably 0.2 to 0.5% by weight of peptone is further added to sterilize the extract. To this, 25 g / L of black yeast ( Aureobasidium pullulans ) can be inoculated and cultured. The culture can be carried out using a 5 L fermenter for 1 to 7 days at 20 to 40 ° C while maintaining the pH at 5-7. After culturing, the culture broth was centrifuged and the culture was firstly removed and sterilized (121 ° C, 15 minutes, 1.5 atm) to prevent further culture.

After the fermentation, the fermented product obtained by removing the cultures firstly is added with ethanol so as to be a final 70% (v / v) ethanol solution, refluxed, cooled, and filtered. When the filtration is completed, the extract obtained in the extracting step is transferred to a concentration tank and concentrated under reduced pressure or freeze-dried at 50 ° C or lower to prepare the fermented extract of Sanjia wood black yeast of the present invention.

According to a preferred embodiment of the present invention, the fermented extract of Sanjay Black yeast added to the cosmetic composition is 0.0001 to 80.0% by weight, preferably 0.01 to 50.0% by weight, and most preferably 0.1 to 50.0% 30.0% by weight. When the content of the above fermented extract is less than 0.0001% by weight, the effect of antioxidation, anti-inflammation, promotion of collagen synthesis, improvement of skin wrinkles, moisturizing, whitening and skin barrier improvement is insignificant.

Meanwhile, the cosmetic composition of the present invention may contain, as an active ingredient, components commonly used in cosmetic compositions in addition to the above-described fermented extract of Sanjay wood black yeast, and may contain components such as antioxidants, stabilizers, solubilizers, vitamins, A conventional adjuvant, and a carrier.

Meanwhile, the cosmetic composition of the present invention may be prepared in any form conventionally produced in the art, and examples thereof include a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, Containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. However, the present invention is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a convergent 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 cosmetic composition of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide Can be used.

When the formulation of the cosmetic composition 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, Propane / butane, or dimethyl ether. ≪ RTI ID = 0.0 >

When the formulation of the cosmetic composition 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 , Propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the cosmetic composition of the present invention is in the form of a suspension, the carrier component may include water, a liquid diluent such as ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester Suspending agent, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the cosmetic composition of the present invention is a cleansing composition containing an interfacial active agent, it is preferable to use, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, Fatty acid amide ether sulfate, alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

On the other hand, the fermented extract of Sanjia wood black yeast according to the present invention has an antioxidant effect (see Experimental Example 2), anti-inflammatory effect (see Experimental Example 4), collagen synthesis effect (see Experimental Example 5) Example 6). In addition, in the case of the cosmetic composition prepared by containing the fermented extract of Sanjay wood black yeast having such an effect as an active ingredient, it was confirmed that the cosmetic composition having an excellent skin moisturizing effect (Experiment Example 7) and skin barrier improving effect (Experiment Example 8) Respectively.

Hereinafter, the present invention will be described in more detail with reference to the following Production Examples. However, the following Production Examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to the following Production Examples.

[ Manufacturing example  One: Sanjay  Preparation of extract]

The fruit of Sanjay was washed with purified water, dried and then crushed, and finely ground with a sieve of 100 mesh. 70% ethanol was added to 100 g / L of acid millet, and the mixture was refluxed three times for 5 hours. After cooling, the mixture was filtered through a filter of whatman # 3, concentrated under reduced pressure at 50 캜 or below, The extracts of Sanjogi trees were prepared.

[ Manufacturing example  2: Sanjay Black yeast  Preparation of fermented extract]

2 g of glucose was added as a carbon source to 5 g of the extract of Sanjay tree prepared in Preparation Example 1, and 0.5% of peptone was further added to sterilize. To this, 25 g / L of black yeast ( Aureobasidium pullulans ) were inoculated and cultured. Cultures were cultured in a 5 L fermenter for 7 days at 37 ° C and pH 5-7. After culturing, the culture broth was centrifuged and the culture was firstly removed and sterilized (121 ° C, 15 minutes, 1.5 atm) to prevent further culture.

After the fermentation, the fermented product of the acidophilus, from which the culture was firstly removed, was refluxed three times for 5 hours with the addition of ethanol so as to be a final 70% (v / v) ethanol aqueous solution. After cooling, Filtered. When the filtration is completed, the extract obtained in the extraction step is transferred to a concentration tank and concentrated under reduced pressure or freeze-dried at 50 ° C or lower.

[Comparative Production Example 1]

It was added a 2% Four clique as a carbon source, and sterilized by adding an additional 0.5% peptone. To this, 25 g / L of black yeast ( Aureobasidium pullulans ) were inoculated and cultured. Cultures were cultured in a 5 L fermenter for 7 days at 37 ° C and pH 5-7. After culturing, the culture broth was centrifuged and the culture was firstly removed and sterilized (121 ° C, 15 minutes, 1.5 atm) to prevent further culture.

After the fermentation, the cultures were firstly removed, ethanol was added to make a final 70% (v / v) ethanol aqueous solution, refluxed three times for 5 hours, cooled, and filtered through Whatman # 3 filter paper. When the filtration was completed, the filtrate was transferred to a concentration tank and concentrated under reduced pressure or freeze-dried at 50 ° C or lower to prepare a black yeast fermentation broth without the extract of Liliaceae.

[ Manufacturing example  3 to 6]

The fermented hornblende extract was prepared in the same manner as in Preparation Example 2, except that the mycelium of Table 1 was used instead of the black yeast.

mycelium Production Example 3   Saccharomyces cerevisiae Production Example 4 Lactobacillus casei Production Example 5 Asperillus oryzae Production Example 6 Bacillus subtilis

[ Experimental Example  1: Vitamin C analysis]

The content of vitamin C in the extract of Sanjay tree of Preparation Example 1 and the fermented extract of Prussian blueberry yeast of Preparation Example 2 was analyzed by HPLC (mobile phase: 25 mM potassium phosphate (KH2PO4), pH 2.5). The analytical column was analyzed with OptiPack C18, column temperature 30 ° C and ultraviolet detector absorbance wavelength 254 nm.

The results of the analysis of the vitamin C content of the extracts of the wild plants of the black yeast fermentation (a) and the black yeast fermentation (b) are shown in FIG. Table 2 shows the vitamin C contents of the fermented extracts of Sanjay Black yeast fermentation extracts of Production Example 2 and Fermented Extracts of Sanjay Woods of Production Examples 3 to 7.

The content of vitamin C before and after fermentation Before fermentation (ppm) After fermentation (ppm) Production Example 2 (Black yeast fermentation) 2,803.4 18,292.5 Production Example 3 (yeast fermentation) 2,803.4 2,805.43 Production Example 4 (fermentation of lactic acid bacteria) 2,803.4 2,803.21 Production Example 5 (fermentation of M. tuberculosis) 2,803.4 2,805.41 Production Example 6 (Bacillus subtilis fermentation) 2,803.4 2,803.25

The content of vitamin C before and after the black yeast fermentation of the extract of Sanjayama was 2803.4 ppm before fermentation, but it was 18,292.5 ppm after fermentation (Production Example 2). However, in the case of other mycelial fermented products, no significant increase in the content of vitamin C was observed.

[ Experimental Example  2: Sanjay  Extract and Sanjay Black yeast  Fermented extract DPPH Evaluation of antioxidant activity by the method]

Free radical scavenging activity was evaluated according to the concentrations (31.5, 62.5, 125, 250, 500, 1000 ㎍ / ml) of the fermented extract of Sanjay wood black yeast obtained in Preparation Examples 1 to 6 and the extract of Sanjay Respectively.

The DPPH assay measures absorbance at 540 nm to the extent that the inhibitor decolorizes the stable radical DPPH (2,2-Diphenyl-1-picrylhydrazyl radical). The used samples are shown in Table 3 below, and the free radical scavenging activity was measured using the following equation (1). The control group was Epigallocatechin gallate (EGCG).

material Control group Blank of C Exp. Group Blank of E A DPPH (MeOH) 180 (180) 180 (180) B Sample (solvent) (20) (20) 20 20

A: Abs of experimental group

B: Abs blank of experimental group

C: Abs of control group

D: Abs of blank of control group

Free radical scavenging activity (%) Production Example 1 Comparative Preparation Example 1 Production Example 2 Production Example 3 Production Example 4 Production Example 5 Production Example 6 EGG 25uM - - - - - - - 54.34 0.001 wt% 12.23 4.93 42.15 22.13 15.54 13.13 21.19 - 0.002 wt% 22.16 6.09 68.24 30.09 22.19 24.17 28.42 - 0.005 wt% 24.58 8.12 83.41 38.42 31.41 33.21 35.64 - 0.01 wt% 35.24 9.09 95.58 55.19 41.28 44.42 45.24 -

As shown in Table 4, the extracts of Fermented Sanjogami (Preparation Examples 2 to 6) were superior to the extracts of Sanjayaki (Preparation Example 1) in free radical scavenging activity. Among fermented extracts, black yeast fermented extract (Preparation Example 2) showed the highest free radical scavenging activity. In addition, superior antioxidative effect was confirmed in comparison with Comparative Production Example 1 including only black yeast fermentation broth. Therefore, it was confirmed that the antimicrobial fermented extract of Sanjia wood black yeast according to the present invention has an excellent antioxidative effect as compared with the extract from Sanjayu.

[ Experimental Example  3: Sanjay  Extract and Sanjay  Evaluation of cell viability of fermented extract]

Toxicity to the cells of the extracts of Sanjay and Sanjayu fermentation extracts obtained in Production Examples 1 to 6 and Comparative Production Example 1 was measured. Cytotoxicity was performed by modifying Mosmann's method of measuring cell viability using 3- (4,5-dimethylthiazol-2-yl) -2-5-diphenyltetrazolium bromide (MTT) reagent.

HDF was dispensed into 96-well plates at a concentration of 1 x 10 ⁴ cells / well and cultured at 37 ° C and 5% CO ₂ for 24 hours. The culture medium was removed and the sample was cultured for 24 hours in a concentration-treated medium. The medium was removed and washed twice with PBS (phosphate buffered saline). MTT was dissolved in PBS at 5 mg / mL, and 50 μL was added and cultured at 37 ° C and 5% CO ₂ for 2 hours. DMSO was added to each well (100 μL), stirred for 10 minutes, and then absorbance was measured at 540 nm.

The measurement results are shown in Table 5 below. As shown in Table 5, no cytotoxicity was observed at all concentrations of each sample. From this, it can be confirmed that the extract of Fermented Sanjay Black yeast according to the present invention is harmless to human body and has excellent safety.

Cell survival rate (%) Name of sample Treatment concentration (ppm) Cell survival rate (%) Production Example 1
50 101.3 100 103.5 Comparative Preparation Example 1
50 99.8 100 101.6 Production Example 2 50 103.5 100 102.5 Production Example 3
50 98.7 100 100.5 Production Example 4
50 99.8 100 103.5 Production Example 5
50 97.5 100 98.6 Production Example 6
50 97.6 100 101.5

[ Experimental Example  4: Sanjay  Extract and Sanjay  Evaluation of anti-inflammation of fermented extract (5- Lipoxygenase  Inhibition)]

To evaluate the anti-inflammatory effect of 5-Lipoxygenase inhibitory activity of the extracts of Fermented Sanjota tree and Sanjay wood extracts obtained in Preparation Examples 1 to 6 and Comparative Preparation Example 1.

Lipoxygenase produces a variety of substances from arachidonic acid that act as various chemical mediators involved in inflammatory and allergic reactions in vivo. Therefore, a substance that inhibits lipoxygenase may result in inhibiting the production of various chemical mediators, so it can be assumed that it is effective for allergy. The activity level of lipoxygenase is an experiment to measure the degree of peroxide formation using substrate and enzyme.

The used samples were tested as shown in Table 6 below, and the 5-lipoxygenase inhibitory activity (%) was measured using the equation (1). The control group was nordihydroguaiaretic acid (NDGA).

material Control group Blank of C Exp. Group Blank of E A Buffer solution 2ml 2ml 2ml 2ml B Sample (solvent) (20 [mu] l) (20 [mu] l) 20ul 20ul C Enzyme 40ul - 40ul - D Substrate 70ul 70ul 70ul 70ul

Production Example 1 Comparative Preparation Example 1 Production Example 2 Production Example 3 Production Example 4 Production Example 5 Production Example 6 NDGA 250 nM - - - - - - - 51.55 0.001 wt% 19.22 3.25 33.19 25.41 21.51 21.39 35.01 - 0.002 wt% 28.49 5.03 41.84 32.01 35.21 34.15 32.54 - 0.005 wt% 33.16 5.42 62.30 34.61 36.76 38.69 32.92 - 0.01 wt% 32.90 5.32 74.43 49.53 41.43 43.04 41.85 -

As shown in the results of Table 7, the extracts of Fermented Sanjota (Production Examples 2 to 6) were superior to the extracts of Liliaceae (Production Example 1) and Comparative Production Example 1 in the inhibition of 5-lipoxygenase. Among the fermented extracts, the extracts of Fermented Blackberry yeast (Preparation Example 2) showed the highest 5-lipoxygenase inhibitory ability. In general, the higher the inhibitory effect of 5-lipoxygenase is, the more anti-inflammatory effect is obtained. Therefore, the above results confirm that the fermented extract of Sanjia wood black yeast has a higher anti-inflammatory effect than the common extract of Sanjogami.

[ Experimental Example  5: Sanjay  Extract and Sanjay  Evaluation of collagen synthesis amount of fermented extract]

The rate of increase in collagen biosynthesis of the extracts of Fermented Sanjota tree and Sanjay wood extracts obtained in Preparation Examples 1 to 6 and Comparative Preparation Example 1 was evaluated.

HDF was added to a 24-well plate at a concentration of 5 x 10 ⁴ cells / well and cultured at 37 ° C and 5% CO ₂ for 24 hours. Serum-free DMEM medium supplemented with the extracts prepared in Preparation Examples 1 to 6 and Comparative Preparation Example 1 and the control group were further cultured for 24 hours in a serum-free DMEM medium containing no extracts from the extracts of Japanese corn tree . 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 / ml, Respectively. Collagen biosynthesis increase rate (%) was calculated according to the following formula (2), and the control group was L-ascorbic acid (L-AA).

Production Example 1 Comparative Preparation Example 1 Production Example 2 Production Example 3 Production Example 4 Production Example 5 Production Example 6 L-AA 0.3 mM - - - - - - - 131.51 0.001 wt% 101.03 101.22 126.65 107.10 114.15 105.96 113.84 - 0.002 wt% 106.15 102.54 162.38 115.03 115.36 114.47 126.37 - 0.005 wt% 115.64 103.65 174.44 120.60 121.52 123.24 129.61 - 0.01 wt% 116.22 103.74 182.35 127.48 131.64 125.82 133.52 -

As can be seen from the results of Table 8, the extracts of Fermented Sanjota Wood (Preparation Examples 2 to 6) of the present invention were excellent in the amount of collagen synthesis as compared to the extracts of Production of Lumber (Preparation Example 1) and Comparative Preparation Example 1. Among the fermented extracts, black yeast fermented extract (Preparation Example 2) showed the highest amount of collagen synthesis.

Experimental Example  6: Measurement of inhibition of melanin synthesis by B16F1 melanocytes

In Experimental Example 6, arbutin known as a whitening agent was used as a comparative sample and B16F1 melanocyte was used to confirm the melanin synthesis inhibitory effect of the samples obtained in Production Examples 1 to 6 and Comparative Production Example 1.

The B16F1 melanocyte used in Experimental Example 6 is a cell line derived from a mouse and is a cell that secretes a melanin pigment called melanin. The inhibitory effect of B16F1 melanin on melanin synthesis was measured as follows. B16F1 melanocytes were plated at a concentration of 2 x 10 ⁶ per well in a 6-well plate, and the cells were adhered, treated at a concentration not causing toxicity, and cultured for 72 hours. After culturing for 72 hours, 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 Rotan (R Lotan and D Lotan, Cancer Res, 40: 3345-3350, 1980). Cell pellets were washed once with PBS and then 1 ml of homogenization buffer (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. Melanin was quantified The percent inhibition of melanin formation in the sample was measured. The inhibition rate (%) of melanin production of B16F1 melanocytes was calculated according to Equation (3).

Melanin synthesis inhibition rate (%) = [(A-B) / A] 100

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

Production Example 1 Comparative Preparation Example 1 Production Example 2 Production Example 3 Production Example 4 Production Example 5 Production Example 6 Arbutin 0.2 wt% - - - - - - - 131.51 0.001 wt% 5.03 3.22 46.65 7.10 5.15 5.96 13.84 - 0.002 wt% 10.10 11.54 62.38 5.03 15.36 14.47 26.37 - 0.005 wt% 15.64 20.65 74.44 16.60 21.52 23.24 29.61 - 0.01 wt% 23.22 23.74 82.35 17.48 22.64 25.82 33.52 -

The inhibitory effect of melanin on B16F1 melanocytes was measured and the results are shown in Table 9. < tb > < TABLE > The fermented extract of Sanjia wood black yeast prepared in Preparation Example 2 showed superior efficacy to arbutin. It was confirmed that it had a better whitening effect than the other production examples.

From the above results, it was confirmed that the melanin synthesis inhibition rate of the fermented extract of Sanjia wood black yeast was excellent, and the excellent whitening effect of the composition for external application for skin of the present invention was confirmed based on this.

[ Manufacturing example  7: Sanjay Black yeast  Fermented extract Cosmetic  Produce]

The cosmetic composition containing 20.0% by weight of the fermented extract of Sanjay wood black yeast of Preparation Example 1 was prepared in the composition shown in Table 10 below, referred to as Prescription Example 1, A cosmetic composition containing a black yeast fermentation broth was prepared and referred to as Comparative Prescription Example 2.

ingredient Prescription Example 1
(weight%) Comparative Prescription Example 1
(weight%) Comparative Prescription Example 2
(weight%) Production Example 1 - 20.0 - Production Example 2 20.0 - - Comparative Preparation Example 1 - - 20.0 1,3-BG 10.0 10.0 10.0 glycerin 5.1 5.1 5.1 Propylene glycol 4.2 4.2 4.2 Tocopheryl acetate 3.0 3.0 3.0 Liquid paraffin 4.6 4.6 4.6 Triethanolamine 1.0 1.0 1.0 Squalane 3.1 3.1 3.1 Macadamia nut oil 2.5 2.5 2.5 Polyahyuvait 60 1.6 1.6 1.6 Azithromycin 1.6 1.6 1.6 Propylparaben 0.6 0.6 0.6 Carboxyl vinyl polymer 1.5 1.5 1.5 incense a very small amount a very small amount a very small amount antiseptic a very small amount a very small amount a very small amount Purified water Balance Balance Balance system 100 100 100

[ Experimental Example  7: Sanjay Black yeast  Skin of fermented extract Moisture power  Improvement effect]

The following experiment was carried out to examine the skin moisturizing effect of the cosmetic composition containing the fermented extract of Saccharomyces cerevisiae of the present invention. Twenty patients in the 20s to 40s without skin diseases were divided into two groups each consisting of 20 patients per group. The nutritional creams of the cream of Prescription Example 1 and Comparative Prescription Example 1 prepared in Preparation Example 7 were divided into two groups each day. For one month. The skin conductivity was measured by using a corneometer (CM820 courage Khazaka electronic GmbH, Germany) under the constant temperature and humidity conditions (24 ° C, 40% humidity) before the start of application, and the skin conductivity was measured at 1 week, 2 weeks, The rate of increase in skin conductivity (%) was measured and the rate of increase was evaluated. The results are shown in Table 11 below.

sample After 1 week after 2 weeks After 4 weeks Prescription Example 1 52 56 61 Comparative Prescription Example 1 28 35 39 Comparative Prescription Example 2 16 19 25

As shown in the results of Table 11, in the case of Formulation Example 1, which is a cosmetic containing the fermented extract of Sanjay Blackberry yeasts of the present invention, the rate of skin conductivity increase was much higher than Comparative Prescription Examples 1 and 2. Since the skin conductivity is generally proportional to the skin moisture content, the above results confirm that the cosmetic composition containing the fermented extract of Sanjay wood black yeast keeps the skin moisture content higher than that of the cosmetic composition not containing it.

[Experimental Example 8: Effect of fermented black tea yeast extract on skin barrier function improvement]

The following experiment was conducted to examine the skin barrier function improving effect of the cosmetic composition containing the fermented extract of the present invention. In this Experimental Example, the cosmetic material of Preparation Example 7 was evaluated by conducting a comparative experiment using a Tewameterr (TM300, Courage and Khazaka Electronic Co., Germany) for transepidermal water loss (TEWL) in humans. Each sample was applied to the right and left faces of 40 women in their 20s and 40s. Percutaneous water loss before, 1, 2, 4, and 6 hours was applied before and 3 cm from the eye. The results are shown in Table 1. The transdermal water loss was measured using a Tewameter (TM300, Courage and Khazaka Electronic Co., Germany) three times and the mean value was evaluated.

division 1 hours 2 hours 4 hours 6 hours Prescription Example 1 15.2 14.8 12.4 9.2 Comparative Prescription Example 1 18.1 17.0 14.2 11.9 Comparative Prescription Example 2 18.7 17.1 15.5 14.6

As shown in Table 12, when the amount of transdermal water loss was measured, the amount of transdermal water loss was reduced in the facial skin of an experimenter who applied cream containing the extract of Fermented Sanjay Black Yeast Extract, .

Claims (10)

A cosmetic composition comprising a black yeast fermented extract of an indigenous fruit. The method according to claim 1,
Wherein the cosmetic composition is for antioxidation. The method according to claim 1,
Wherein the cosmetic composition is for anti-inflammatory use. The method according to claim 1,
Wherein the cosmetic composition is for improving skin wrinkles. The method according to claim 1,
Wherein the cosmetic composition is for skin whitening. The method according to claim 1,
Wherein the cosmetic composition is for moisturizing the skin. The method according to claim 1,
Wherein the cosmetic composition is for improving the skin barrier function. 8. The method according to any one of claims 1 to 7,
The black yeast fermentation extract of the above-mentioned nuts has been obtained by fermenting the extract of Sanjay Natto for 1 to 7 days while maintaining the pH at 5 to 7 under a temperature condition of 20 to 40 ° C as black yeast. . 8. The method according to any one of claims 1 to 7,
Wherein the black yeast fermented extract of the fruit of Sanji is added in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. 8. The method according to any one of claims 1 to 7,
The cosmetic composition may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray Wherein the cosmetic composition has one formulation.

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