KR101855206B1 - Cosmetic Composition Comprising the Fermented Extract of Panax ginseng - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a fermentation broth of cinnamon germ fermented successively. Preferably, the camellia fermentation broth is obtained by secondary fermentation using a primary fermentation broth obtained by cultivating camellia ginseng as a Bacillus subtilis or a yeast (Saccharomyces cerevisiae) using Lactobacillus plantarum. The camellia fermentation broth of the present invention promotes skin cell activity, promotes the synthesis of skin cell proteins such as collagen, inhibits the production of collagenase, thereby improving wrinkles and wounds caused by aging . The ginseng fermentation broth of the present invention has moisturizing and whitening effect.

Description

[0001] The present invention relates to a cosmetic composition comprising a fermentation broth of camphor ginseng (Cosmetic Composition Comprising the Fermented Extract of Panax ginseng)

The present invention relates to a cosmetic composition comprising a ginseng fermentation broth.

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 reaction 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 And has been used as a whitening cosmetic. However, since they are low in stability in cosmetic formulations, their use is limited due to their decomposition and coloring, the generation of odor, the efficacy at the living body level, the uncertainty of the effect, and the 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 stored 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.

As a result of intensive efforts to solve various skin troubles damaged by aging, the inventors of the present invention have found that when a primary fermentation broth fermented with Bacillus subtilis microorganism or yeast bacterium is used as a fermentation solution secondary fermented with lactic acid bacteria, The present inventors have completed the present invention by confirming that the problems of the skin, particularly the skin wrinkles, can be effectively improved. Accordingly, it is an object of the present invention to provide a cosmetic composition having an excellent skin wrinkle-reducing effect. It is another object of the present invention to provide a cosmetic composition having skin elasticity improvement, moisturizing and whitening effect.

The above-mentioned problem is attained by a cosmetic composition comprising an effective fermentation solution of camellia seeds which has been continuously fermented.

Preferably, the camellia fermentation broth is a fermentation broth comprising primary fermentation of camellia ginseng with a strain of Bacillus subtilis or yeast, wherein the primary fermentation broth is selected from the group consisting of Bifidus bacteria, Bulgarian bacterium, Lactobacillus It may be a fermentation liquid of bacillus-lactic acid bacteria or a yeast-lactic acid fermentation broth obtained by secondary fermentation using one kind of lactic acid bacteria strain.

Also preferably, the strain of Bacillus subtilis is Bacillus subtilis, and the yeast may be Saccharomyces cerevisiae.

In addition, the cosmetic composition has an effect of improving skin wrinkles, improving skin elasticity, moisturizing or whitening.

Preferably, the camellia fermentation broth may be contained in an amount of 0.0001-30.0% (w / w) based on the total weight of the cosmetic composition.

The cosmetic composition of the present invention comprises a tea germ extract as an active ingredient and the ginseng fermentation broth is a fermentation broth of yeast-lactic acid fermentation broth or a bacterium-lactic acid bacterium which promotes cellular activity and collagen synthesis, It has an effect of effectively inhibiting the biosynthesis of the enzyme (MMP-1). The ginseng fermentation broth according to the present invention alleviates skin cell stimulation by external stimulation, brightens skin, and has a skin moisturizing effect.

In order to achieve the above object, the present invention provides a cosmetic composition for improving skin wrinkles comprising a ginseng fermentation broth.

In the present invention, Panax Ginseng is a kind of wild ginseng, and the wild ginseng is classified into three kinds of ginseng, geomorphic, and camphor (racial). The ginseng is natural wild ginseng, which algae excrete in the mountains after eating seeds, The seeds of a thousand species have fallen around, and the ginseng ginseng refers to wild ginseng that has been cultivated in a wild state by artificially sowing the seeds of the wild ginseng in deep mountains. Among these wild ginsengs, it is known that the relatively easy to purchase ginseng extract has higher saponin and pharmacological effect than the ginseng. The known pharmacological efficacy of camphor ginseng is the treatment of diabetes, liver disease, hypertension, gynecological diseases and menopausal symptoms.

In the present specification, the term " camellia ginseng " refers to various organs (e.g., leaves, fruits, flowers, stems and roots) of camphor ginseng, and preferably means roots of camphor ginseng.

As used herein, the term " camellia fermentation broth obtained by sequential fermentation "refers to a fermented bacillus-lactic acid fermentation broth obtained by culturing a lactic acid bacterium strain in a primary fermentation broth fermented using a Bacillus sp. Strain or fermented with yeast Refers to a yeast-lactic acid fermentation broth obtained by cultivating a lactic acid bacterium strain in a second fermentation broth.

In the present invention, the Bacillus strain is a Gram-positive microorganism, capable of endogenous spore formation, and has a rod shape. The Bacillus strain useful for primary fermentation of camellia ginseng is Bacillus subtilis.

In addition, the yeast of the present invention belongs to a eukaryotic microorganism (a nuclear and cytoplasmic organism separated by a nuclear membrane) and has cell organelles such as a cell wall, a cell membrane, a nucleus, a mitochondria, and a granule. Those with no distinction of sex mainly reproduce by budding. Among many separated yeasts, there is no efficacy, fission instead of budding or bud-fission, ballistospore, not ascospore, Or spores (spores) are not observed in the yeast. The size is usually 5 ^-10 × 5 ^-12 μm, 5 ^-10 times the size of bacteria, and the optimum pH is 4.5 to 5.5 or 1 to 10 survivors. Yeast surviving at pH 1 for 12 hours is stronger in acid than in bacteria, so if the pH of the medium is adjusted to about 3.5 to 3.8, bacterial contamination can be suppressed to some extent.

Yeast is slightly different depending on species, but most of them grow well at 20 ~ 30 ℃. It is usually said that yeast is killed at temperatures above 50 ° C, but it can be overcome to some extent by adding growth factors that are not synthesized above that temperature, directly to the medium. In the case of Saccharomyces cerevisiae, it can be prevented by adding oleic acid and ergosterol to the medium at a temperature of 40 ° C. or higher. Yeast cells themselves are unicellular plants and contain large amounts of proteins and trace minerals.

The yeast useful for primary fermentation of camellia seeds of the present invention is Saccharomyces cerevisiae.

In the present invention, the lactic acid bacteria strain used in the second fermentation is a bacterium that produces lactic acid by fermenting sugars and is gram-positive, does not have spores, and is micro-aerobic or anaerobic. Lactic acid bacteria include acidophilus, which is useful for the production of lactic acid bacteria, bifidus bacteria which live in the field of breastfed babies, and bacteria which make fermented products called Bulgarian milk. .

The lactic acid bacterium useful for secondary fermentation of the camellia ginseng according to the present invention is Lactobacillus plantarum.

In the present invention, the successively fermented camellia fermentation broth can be prepared by a method comprising the following steps:

a) finely dividing the ginseng ginseng powder using 100 to 500 mesh;

b) adding 1 to 50 g / L of the camphor ginseng powder prepared in step a) to a conventional microorganism culture solution, adding Bacillus strains or yeasts in an amount of 10,000 to 100,000 cfu / L;

c) culturing the culture of step b) at 30 to 37 DEG C for 5 to 10 days under aerobic or anaerobic conditions at a pH of 5 to 7;

d) vacuum sterilizing the culture of step c) at 80 ° C to 85 ° C;

e) filtering the sterilized culture solution of step d), aging the solution at low temperature for 5 to 10 days and filtering;

f) lyophilizing the filtrate of step e) to obtain a powder;

g) adding lactobacillus strain to the lactic acid bacteria medium in an amount of 100,000 to 1,000,000 cfu / L;

h) 1 to 30 g / L of the fermentation broth of step f) is added to the lactic acid bacterium culture of step g) and cultured at 30 to 37 DEG C for 3 to 6 days at pH 3-6 for 5 to 10 days under mooring or anaerobic conditions ;

i) vacuum sterilizing the culture of step h) at 80 ° C to 85 ° C;

j) filtering the sterilized culture solution of step i), aging the solution at low temperature for 5 to 10 days, and filtering;

k) lyophilization to obtain a ginseng fermentation broth powder.

According to a preferred embodiment of the present invention, the ginseng fermentation broth used in the present invention was obtained as follows:

The ginseng powder was pulverized to 300 mesh, added with 10 g / L to the culture medium, added with 500,000 cfu / L of Bacillus strain or yeast strain and cultured at 30 ~ 37 ℃ under aerobic or tuberous anaerobic conditions And cultivated in a fermenter for about 7 days. After culturing, the resulting culture is vacuum-sterilized at 85 ° C, filtered through 0.25 μM filter paper, and aged at low temperature for 7 days. The resulting filtrate was lyophilized to obtain a powder of yeast fermentation broth. The powder was added to the lactic acid bacteria medium at a concentration of 5 g / L, and the lactobacillus strain was added at 1,000,000 cfu / L. Or anaerobic conditions in a fermenter for about 7 days. The culture was vacuum-sterilized at 85 ° C, filtered through a filter paper of 0.25 μM, aged at low temperature for 7 days, filtered and lyophilized to obtain a ginseng fermentation broth. The ginseng fermentation broth was maintained at a final solid concentration of 10 g / L before use

In the present invention, the fermentation broth of Camellia sinensis promotes cell activity, promotes collagen synthesis, inhibits the production of collagenase, improves moisturization, improves wrinkles, brightens skin, alleviates skin irritation It has a variety of features. Accordingly, the present invention shows very good effects on skin damage caused by aging, i.e., skin wrinkles, reduced elasticity, whitening, and moisturizing, by adding a camellia fermentation broth to a cosmetic.

In the present invention, the content of the camellia fermentation broth is 0.0001 to 30.0% (w / w), more preferably 0.001 to 10% (w / w) based on the total cosmetic composition. When the content of camellia fermentation broth is less than 0.0001% (w / w), it is not expected to have a remarkable effect of improving wrinkles, improving elasticity, moisturizing and whitening effect. When the content of camellia fermentation broth exceeds 30.0% There is no increase in the effect, and it causes skin irritation or problems in the stability of the formulation.

The ingredients contained in the cosmetic composition of the present invention include components commonly used in cosmetic compositions in addition to the camellia fermentation broth as an active ingredient and include conventional additives such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, and Carrier.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion (skin), a nutritional lotion (milk 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 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 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 an interfacial active agent-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 sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing agent, or a surfactant-free cleansing agent, it may be applied to the skin and then wiped off or removed or washed with water. The surfactant-containing cleansing formulation is 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.

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 .

compare Manufacturing example  1. Preparation of yeast fermentation broth using yeast

After washing with purified water and drying, the dried ginseng ginseng was pulverized and made fine using 300 mesh. This was added to a yeast culture solution (Potato Dextrose Broth, Akyumedia, USA) at 10 g / L. Saccharomyces cerevisiae was used as a yeast strain used for fermentation and added at 500,000 cfu / L per culture. The cultures were maintained at 30 ° C and pH 5-7 for 7 days using a 5L fermenter. After incubation, the culture broth was vacuum-sterilized at 85 ° C, centrifuged, and the culture was firstly removed, followed by final filtration using 0.25 μM filter paper. The filtrate thus obtained was aged at a low temperature warehouse at 4 ° C for about 10 days, and then the filtrate was freeze-dried by using a 0.25 μM filter to obtain a 'ginseng yeast fermentation broth' powder.

compare Manufacturing example  2. Bacillus  Used ginseng Bacillus  Manufacture of fermentation broth

After washing with purified water and drying, the dried ginseng ginseng was pulverized and made fine using 300 mesh. This was added to a 10 g / L Bacillus culture (Tryptic soy broth, Acumedia, USA). Bacillus subtilis (Bacillus subtilis) was used for fermentation and added at 1,000,000 cfu / L per culture. Cultures were cultured in a 5 L fermenter for 7 days at 37 ° C and pH 5-7. After incubation, the culture broth was vacuum-sterilized at 85 ° C, centrifuged, and the culture was firstly removed, followed by final filtration using 0.25 μM filter paper. The filtrate thus obtained was aged at a low temperature warehouse at 4 ° C for about 10 days, and then the filtrate was lyophilized using a 0.25 μM filter to obtain a 'Chennai ginseng bacillus fermentation broth' powder.

Manufacturing example  1 and 2. Preparation of fermented Ganoderma lucidum using lactic acid bacteria

Powder of the 'ginseng yeast fermentation broth' or the 'ginseng extract of Bacillus cereus Bacillus' in Comparative Preparation Examples 1 and 2 was added to a lactic acid bacteria culture solution (MRS broth, Difco, USA) so as to have a concentration of 5 g / L. Lactobacillus plantarum was used as a lactic acid bacteria strain for fermentation and added at 1,000,000 cfu / L per culture. Cultures were cultured in a 5 L fermenter for 7 days at 37 ° C and pH 3 to 6. After incubation, the culture broth was vacuum-sterilized at 85 ° C, centrifuged, and the culture was firstly removed, followed by final filtration using 0.25 μM filter paper. The resulting filtrate was aged for about 10 days at a low temperature warehouse at 4 DEG C and the filtrate was lyophilized by using a 0.25 mu M filter to obtain a solution of 'Jangbyeonggam-yeast-lactic acid fermentation broth' (Preparation Example 1) - > Lactic acid fermentation broth (Preparation Example 2). Generally, the above Production Example 1 and Production Example 2 are referred to as a cinnamon germ fermentation broth.

Experimental Example  1: of ginseng fermentation broth MMP -1 production inhibitory effect

Using the samples obtained in Preparation Examples 1 and 2 and Comparative Preparation Examples 1 and 2, the following experiment was conducted. First, normal human fibroblasts were inoculated into each well of a 48-well microplate (Nunc, Denmark) at 1 × 10 ⁶ cells and cultured in DMEM medium (Sigma, USA) at 37 ° C. for 24 hours. Next, experiments were conducted in which the concentrations of Preparation Examples 1 and 2 or Comparative Preparation Examples 1 and 2 were changed to serum-free DMEM medium at 0.1% and 0.5% (wt%), respectively, The control group, which was replaced with serum-free DMEM medium, was further cultured for 48 hours.

After the incubation, the supernatant of each well was collected and the amount of newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, USA), and the amount of MMP-1 was converted to ng / ml. TGF-beta (10 ng / ml, Roche, USA) was used as a positive control and the inhibition rate of MMP-1 production was calculated according to the following formula 1.

Test substance concentration (% by weight) Inhibition rate of MMP-1 production (%) Positive control (TGF-β) 10 ng / ml 67.4 Comparative Preparation Example 1 0.1 wt% 24.8 Comparative Preparation Example 1 0.5 wt% 61.8 Comparative Preparation Example 2 0.1 wt% 22.1 Comparative Preparation Example 2 0.5 wt% 59.1 Preparation Example 1 0.1 wt% 59.4 Preparation Example 1 0.5 wt% 89.3 Preparation Example 2 0.1 wt% 54.8 Preparation Example 2 0.5 wt% 85.8

Experimental Example  2: Enhancement of Collagen Synthesis Effect of Ganoderma lucidum Fermentation Liquid

Normal human fibroblasts were inoculated into each well of a 48-well microplate to a concentration of 1 x 10 ⁶ cells and cultured in DMEM medium at 37 ° C for 24 hours. Subsequently, the control group in which Production Examples 1 and 2 and Comparative Preparation Examples 1 and 2 were replaced with serum-free DMEM medium at a final concentration of 0.1 wt% and 0.5 wt%, respectively, and a control group in which serum-free DMEM medium containing no cambium- Were further cultured for 24 hours. After culturing, the supernatant of each well was collected and the amount of procollagen type I C-peptide (PICP) was measured using a kit (Takara, Japan). The amount of PICP was converted to ng / ml, and TGF-β was also used as a positive control. The increase rate of collagen biosynthesis was calculated by the following equation (2) and the results are shown in Table 2.

Test substance concentration (% by weight) Increase in collagen production (%) Positive control (TGF-β) 10 ng / ml 76.6 Comparative Preparation Example 1 0.1 wt% 20.8 Comparative Preparation Example 1 0.5 wt% 62.5 Comparative Preparation Example 2 0.1 wt% 18.7 Comparative Preparation Example 2 0.5 wt% 59.4 Preparation Example 1 0.1 wt% 67.1 Preparation Example 1 0.5 wt% 94.7 Preparation Example 2 0.1 wt% 64.7 Preparation Example 2 0.5 wt% 89.8

Through the above Experimental Examples 1 and 2, it can be seen that the cambium fermentation broth according to the present invention inhibits the biosynthesis of MMP-1 having collagen degradation characteristics and is excellent in promoting collagen synthesis and has a better effect than the fermentation broth of Camellia Bacillus Was much better.

Experimental Example  3: Fibroblast proliferation effect of ginseng fermentation broth

Normal human fibroblasts were inoculated into each well of a 96-well microplate to a concentration of 1 x 10 ⁴ cells and cultured in DMEM medium at 37 ° C for 24 hours. Then, in Experimental Examples 1 and 2 and Comparative Preparation Examples 1 and 2, the experimental group was changed to a DMEM medium containing 2.5% by weight of serum with a final concentration of 0.1% by weight and 0.5% by weight, respectively, The control group, which was replaced with DMEM medium, was further cultured for 24 hours. After incubation, MTT (Sigma, USA) solution (3 mg / ml) was added to compare the cell viability and the cell viability was measured at 570 nm using an ELISA reader (Molecular Devices, USA) The cell viability (%) was calculated according to the following table.

Test substance concentration (% by weight) Cell growth rate (%) The positive control (TGF-β: 10 ng / ml) 131.6 Comparative Preparation Example 1 0.1 wt% 48.8 Comparative Preparation Example 1 0.5 wt% 129.5 Comparative Preparation Example 2 0.1 wt% 39.3 Comparative Preparation Example 2 0.5 wt% 121.3 Preparation Example 1 0.1 wt% 130.3 Preparation Example 1 0.5 wt% 181.7 Preparation Example 2 0.1 wt% 130.7 Preparation Example 2 0.5 wt% 180.8

From the above experimental results, it can be seen that the cambium fermentation broth according to the present invention has a much higher cell growth rate than the fermentation broth of camphor brewer's yeast and camellia Bacillus and exhibits a similar or even better cell growth rate to that of the positive control, TGF-β there was.

Experimental Example  4. Effect of Lactic Acid on Cell Stimulation in Changb

Human fibroblasts (Korean Cell Line Bank, Korea) were cultured in T-75 flasks (Falcon, USA) until they grew to about 80%. The cells were transferred to 96-well plates (Falcon, USA) at 3 × 10 ⁴ cells / well and cultured for 24 hours. After culturing, cells were completely adhered to each other through a microscope to confirm whether they were growing well. Experiments were conducted using lactic acid as a skin cell stimulating agent. At this time, 0.2 wt% of lactic acid was used. That is, 200 쨉 l of DMEM (Sigma, USA) medium containing 0.2% by weight lactic acid was added to each of 96 wells, and Preparation Examples 1 and 2 and Comparative Preparations 1 and 2 were adjusted to 0.1 and 0.5 wt% Respectively. At this time, no lactic acid and camellia fermentation broth added were used as a control group, and only lactic acid supplemented group was used as a comparative group. MTT (Sigma, USA) solution (3 mg / ml) was added to compare cell survival rates after 12 hours of addition of the test substance, and the cell viability was measured at 570 nm using an ELISA reader (Molecular Devices, And cell viability (%) was calculated according to the following equation (4). The experimental results are shown in Table 4 below.

Test substance concentration (% by weight) Cell growth rate (%) Control group 99.8 The comparative group (lactic acid 0.2%) 0.00 Lactic acid (0.2%) Comparative Production Example 1 0.1% 30.8 Lactic acid (0.2%) + Comparative Preparation Example 1 0.5 wt% 68.9 Lactic acid (0.2%) + Comparative Preparation Example 2 0.1% 32.8 Lactic acid (0.2%) + Comparative Preparation Example 2 0.5% 67.9 Lactic acid (0.2%) + Preparation Example 1 0.1% 74.6 Lactic acid (0.2%) + Preparation Example 1 0.5% 91.6 Lactic acid (0.2%) + Preparation Example 2 0.1% 70.5

From the above results, it was found that the treatment of cambium fermentation broth significantly inhibited the cell formation by lactic acid treatment, greatly increased the cell production, and the cell growth rate was much better than that of the fermentation broth of camphor brewer's yeast and Camellia sinensis Bacillus .

Experimental Example 5: Effect of fermented extract of cinnamon gum on inhibition of melanin formation using B16F1 melanin-forming cells

Experimental Example 5 was performed to determine the whitening effect of B16F1 melanocyte-forming cells in order to examine the whitening effect of the fermentation broth. 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 melanogenesis cells were divided into 6 well plates at a concentration of 2 X 10 ⁵ per well and treated with Preparation 1 and 2 and Comparative Preparations 1 and 2 at a concentration that did not induce toxicity after the cells were attached and incubated for 72 hours Respectively. 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 by the following formula (5), and the results are shown in Table 5.

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
(weight%) Melanin formation inhibitory effect (%) Comparative Preparation Example 1 Comparative Production Example 2 Production Example 1 Production Example 2 0.01 7.17 5.75 22.75 22.54 0.05 15.67 16.19 36.19 39.12 0.1 24.14 21.24 57.24 51.54 0.5 39.16 35.01 85.01 84.05 200ppm arbutin 66.15

As can be seen from the results of Table 5, the fermentation broth of the cinnamon produced by Production Examples 1 and 2 of the present invention significantly inhibited melanogenesis in B16F1 melanin-forming cells, and compared with the inhibitory effect of melanin production of Comparative Preparation Example 1 or 2 And it was confirmed that it was excellent.

Formulation Example 1: Preparation of a cosmetic composition containing a fermentation broth of cinnamon ginseng

The cosmetic compositions containing the camellia fermentation broth of Preparation Examples 1 and 2 were prepared as cosmetic compositions (Formulation Examples 1 and 2) containing the fermentation broth of camellia ginseng. In order to compare the efficacy, (Comparative Formulation Example 1), a cosmetic composition comprising glycerin and 1,3-butylene glycol as a moisturizing agent (Comparative Formulation Example 2) instead of a cinnamon fermentation broth, a cosmetic composition containing adenosine as a wrinkle reducing raw material (Comparative Formulation Example 3), and a cosmetic composition (Comparative Formulation Example 4) that does not contain all of the ginseng fermentation broth and moisturizer are shown in Table 6 below.

(Unit: wt%) division ingredient Formulation Example 1 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Comparative Formulation Example 4 A





Ginseng fermentation broth
(Production Example 1) 2.0 - - - - Fermented juice
(Comparative Production Example 1) - 2.0 - - - glycerin - - 5.0 - - 1,3-butylene glycol - - 5.0 - - EDTA-2Na 0.02 0.02 0.02 0.02 0.02 adenosine - - - 0.04 - Purified water to 100 to 100 to 100 to 100 to 100 B









Cetostearyl alcohol 2.0 2.0 2.0 2.0 2.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 Microcrystalline 0.7 0.7 0.7 0.7 0.7 Squalane 5.0 5.0 5.0 5.0 5.0 Liquid paraffin 3.0 3.0 3.0 3.0 3.0 Trioctanoin 5.0 5.0 5.0 5.0 5.0 Polysorbate 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3.0 3.0 3.0 3.0 3.0 BHT 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

Experimental Example  6: Moisturizing effect of Ganoderma lucidum fermentation broth

The clinical moisturizing effect of the cosmetic composition of Formulation Example 1 was measured as follows. 100 healthy adults and women who feel dry skin were divided randomly into four groups (A, B, C and D) by 25 persons, and the cosmetic composition of Formulation Example 1 was compared to the composition of Group A The cosmetic composition of Formulation Example 1, the cosmetic composition of Comparative Formulation Example 2, and the cosmetic composition of D Comparative Formulation 4 were applied to the face for 2 weeks, twice a day 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 7 below.

division Before use After 2 weeks of use After 4 weeks of use Formulation Example 1 23.1 39.5 54.2 Comparative Formulation Example 1 24.3 29.1 35.9 Comparative Formulation Example 2 23.5 36.3 48.7 Comparative Formulation Example 4 23.7 24.4 24.2

As shown in Table 7, it was confirmed that the moisturizing effect of the cosmetic composition (Formulation Example 1) containing the fermented ginseng fermented broth of the present invention of the present invention is superior to the cosmetic composition containing the other moisturizing agents (Comparative Formulation Example 2).

Experimental Example 7. Improvement of Skin Wrinkle of Root Bran Ginseng Fermentation Solution

       Clinical trials were conducted on the effect of the cosmetic composition containing the fermentation broth of camellia ginseng according to the present invention on the skin wrinkles. Clinical trials were conducted through actual use tests of each cosmetic product. That is, in a clinical experiment, 100 female 40-60 year-olds were randomly divided into four groups (A, B, C and D) by 25 persons. The cosmetic composition of Formulation Example 1 was applied to Group A, 1, the cosmetic composition of Comparative Formulation Example 2, and the cosmetic composition of Comparative Composition D 3 were applied to the face in the face for 8 weeks twice a day, respectively. The wrinkle improvement effect after 8 weeks was visually evaluated to confirm the wrinkle improvement degree. The results of the wrinkle-improving effect based on this evaluation are shown in Table 8 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 8 below.

Wrinkle improvement effect Effective rate (%) Great slightly none Formulation Example 1 19 4 2 92.0 Comparative Formulation Example 1 5 7 13 48.0 Comparative Formulation Example 2 4 4 17 32.0 Comparative Formulation Example 3 15 6 4 84.0

As can be seen from the results of Table 8, the wrinkle-improving effect was about 1.9 times higher than that of Comparative Formulation Example 1 containing the fermentation broth of Form 1 of formulation 1 containing the fermentation broth of cambium of the present invention. In addition, similar results were obtained with the cosmetic preparation of Comparative Example 3 containing adenosine, which is known to have a wrinkle-improving effect instead of the camphor ginseng fermentation broth, indicating that the camellia fermentation broth exhibits excellent wrinkle-improving effect.

Experimental Example  8: Measurement of skin elasticity improvement effect

In Experimental Example 8, to evaluate the skin elasticity improvement effect of the cosmetic composition according to Formulation Example 1 and Comparative Formulation Example containing the fermented solution of cinnamon ginseng obtained in Preparation Example 1, 30 women with 30 to 40 elasticity decreased Respectively. 10 groups were divided into 4 groups of A, B, and C, followed by Formulation Example 1 for Group A, Comparative Formulation Example 1 for Group B, and Comparative Formulation Example 4 for Group C for 12 weeks The 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 9 below, and the results are an average value of improvement for each group.

Experimental product Skin elasticity improvement (%) Formulation Example 1 40.4 Comparative Formulation Example 1 23.4 Comparative Formulation Example 4 3.6

As shown in Table 8, Formulation Example 1 containing the fermented extract of Camellia sinensis significantly improved skin elasticity as compared to Comparative Formulation Example 1 or 4.

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. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

The ginsenosides were firstly fermented using Saccharomyces cerevisiae , which is a yeast strain, and the first fermentation broth was subjected to a second fermentation using lactobacillus plantarum ( Lactobacillus plantarum ) As an active ingredient. The cosmetic composition according to claim 1, wherein the camellia fermentation broth is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition.

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