KR20150136290A - Extract Of bee venom, extract of willow bark and extract of fermented rice Containing Cosmetic Composition Having Anti-aging Effect - Google Patents

Abstract

The present invention relates to a cosmetic composition for anti-aging, containing, as active ingredients, extracts of bee venom, the bark of willow trees, and fermented rice. The cosmetic composition for anti-aging is excellent in terms of antioxidative effects, effects of biosynthesizing collagen fibers, and effects of inhibiting hyaluronidase activities. Furthermore, the composition is safe on skin and shows excellent effects of improving skin elasticity.

Description

TECHNICAL FIELD The present invention relates to an anti-aging cosmetic composition containing bee venom extract, willow bark extract and rice fermented extract.

The present invention relates to an anti-aging cosmetic composition, and more particularly, to an anti-aging cosmetic composition containing bee venom extract, willow bark extract and rice fermented extract as an active ingredient. The antioxidant effect, the biosynthesis effect of collagen fiber And the effect of inhibiting the activity of hyaluronase. The skin elasticity enhancement effect and skin safety are excellent.

Recently, there has been a lot of interest in skin aging due to the well-being and healing craze. This is because aging of muscles and skeleton, along with aging of skin, is a process that directly affects individual's daily life and psychological and social well-being.

Skin is also the primary sensory organ, the body's primary line of defense against infectious organisms and physical harm, and it also plays an important role in regulating body temperature.

The skin aging mechanism which is the foundation of the development of antioxidant cosmetics is that the skin is continuously exposed to oxidative stress from harmful environment such as air pollution, ultraviolet ray exposure, stress or disease, and the radicals in the body are increased, It can be explained that it collapses collagen (collagen), elastin, hyaluronic acid which are connective tissues and causes settlement phenomenon (wrinkles) in a certain part of the skin. In addition, such skin aging mechanism oxidizes the lipid part of the cell membrane and causes destruction of cells, thereby causing diseases such as dermatitis, acne or skin cancer. In addition, the radicals in the body participate in the melanin formation process, which causes spots, freckles and wrinkles.

Regarding the skin aging mechanism, studies on the systemic and endogenous physiological aging with the increase in age and the local exogenous photoaging by ultraviolet rays exposed for a long time are attracting attention. Especially, face and tofu, which are important parts of cosmetics, are found to be greatly affected by photoaging, and research on photoaging and wrinkling of human skin and animal models is actively under way. The internal structure of the skin changes depending on the photoaging, for example, epidermal, dermal hypertrophy, accumulation of elastin, or change in the three-dimensional structure of elastic fibers. In addition, the microfibrillation of the collagen fibers and the weakening of the fiber shape have been revealed in association with collagen, which is the main constituent skeleton of the skin structure. (Zheng P. and Kligman LH, uvA-induced ultrastructural changes in hairless mouse skin: A comparison However, as for collagen fiber bundles formed by binding collagen fibers, sufficient changes have been made up to now on structural changes due to photoaging, changes in skin physical properties, and the like. It is known only that the skin toughness and the like are affected by the study of the skin wound treatment process and the like.

Recently, research is underway on the direct and indirect effects of sunlight due to destruction of the ozone layer, which has been found to be the mainstream of skin aging research. Particularly, in order to clarify the effect of the sunlight on the formation of wrinkles, various natural ingredients are being actively studied for effect on collagen fibers.

Conventional methods for preventing skin aging or improving skin wrinkles have been to use cosmetic preparations or medicines containing ascorbic acid,? -Tocopherol, retinol and its derivatives, SOD, and the like. However, these cosmetic and pharmaceutical products are not only expensive but also have poor chemical stability, so that it is difficult to expect a substantial effect in pharmaceuticals, foods, and the cosmetics industry.

Recently, interest in natural materials has been increasingly studied. Among these, bee venom, which is a pure natural substance and has a strong antibacterial and antiinflammatory effect, is stored in the money of queen bee or worker beetle. It gradually increases in quantity after birth and becomes about 0.3 mg at 15 days old bees After about 17 days of age to keep rumors, most of the bee venom, which is not produced further, is a protein and peptide ingredient, often measuring the maturity of poison production by the total protein content of the capsular bag. The toxins obtained from bees are very complex mixtures composed of enzymes, polypeptides, and various materials with small molecular weights. About 40 kinds of toxins are known as main components. Among them, representative components are melitin, phospholipase phospholipase, hyaluronidase A 2, histamine, apamin, adolapin, MCD peptide, dopamine, protease inhibitor, (B), α-glucosidase, and lysophospholipase, in particular, the main component of the main component, mellipine Melittin has anti-inflammatory and antimicrobial actions, powerful analgesic action, and immune enhancement. Bee venom has both antibacterial activity against gram negative and positive bacteria, but has stronger effect against gram positive bacteria. Also, bee venom promotes regeneration of skin damaged by ultraviolet rays, and has been used for diseases such as arthritis and gout for a long time because of the incontinence therapy because there is little risk of side effects and residues.

The bee venom of domestic honey bee (Apis mellifera) can be obtained by collecting only a large amount of pure bee venom without damaging the honey bee by using the bee venom collecting device using the electric impact method, and then removing the foreign matter and freeze drying the purified bee venom.

The willow is a shrub and tree belonging to the Salicaceae Willow (Salixaceae). It is the origin of the northern temperate region. It is a deciduous arboreous tree about 10-15㎝ long, with branches hanging downward and small branches brown or reddish brown . Leaves are sharp sawtooth, long end and pointed, flowers bloom in March-April, and the time of fruit is May. Leaves are 柳 葉, flowers are 毛花, fruit is lily of the field, and husky skin is 유 유 유 유. It grows in China, origin in Jeollanam-do, Korea in the country, and also grows in fields of China and Europe. There are three species of willow tree, S. nigra, S. fragilis and S. alba, all of which grow up to over 20㎝ tall. Salix nigra grows in North America, while the other two are native to Eurasia, but now they are growing as a naturalized plant.

Rice is a valuable food resource, one of the world's top three grains, which is half of the world's population, and is a key national resource, especially in Asia, which can not be confronted with other crops. Rice, which is rice husk, which is the surface of rice, is called brown rice, rice rice with rice husk and rice husk is removed and rice is called white rice.

Rice (Oryza sativa) is a first-year primordium belonging to rice paddy and is widely distributed throughout the world. Rice constitutes about 70 ~ 85% of carbohydrates, 6.5 ~ 8.0% of protein, 1.0 ~ 2.0% of fat and about half of the world 's population is used as stock. Rice accounts for about 21% of the calories consumed by humans and is a very important crop grown in over 120 countries. In the case of brown rice, protein, lipid, fiber and ash are distributed in the chaff and colony, and it is known that the content of minerals and vitamins is 2 ~ 3 times as high as that of nutrition.

If the embryo is removed from the embryo, the embryo will be removed from the embryo. If the embryo is left untreated, the embryo will retain its taste, and the protein and lipids contained in the embryo, as well as other essential nutrients such as minerals and vitamins .

Accordingly, the present inventors have repeatedly carried out studies for imparting anti-aging effects by using bee venom extract, willow bark extract, and rice fermentation extract in order to develop a cosmetic composition for preventing skin aging from rapid westernization and environmental pollution As a result, the extracts of bee venom extract, willow bark extract and rice fermentation extract have excellent antioxidative effect, biosynthesis effect of collagen fiber and inhibition of hyaluronase activity. Skin elasticity enhancement effect, and skin safety. The present invention has been completed based on this finding.

Accordingly, an object of the present invention is to provide an anti-aging cosmetic composition comprising bee venom extract, willow bark extract and rice fermented extract as an active ingredient.

In order to achieve the above object, the present invention provides an anti-aging cosmetic composition comprising bee venom extract, willow bark extract, and rice fermented extract as an active ingredient.

The bee venom among the active ingredients in the cosmetic composition for anti-aging of the present invention means a secretion secreted from the bees' innards, and is not particularly limited as long as it is a secretory secretory bee.

The bee venom can be obtained by an electric shock method.

The cosmetic composition according to one aspect of the present invention may contain 0.0001 to 30% by weight of a composition containing bee venom extract as an active ingredient based on the total weight of the composition. The cosmetic composition according to another aspect of the present invention may contain 0.001 to 15% by weight of a composition containing bee venom extract as an active ingredient based on the total weight of the composition. The cosmetic composition according to another aspect of the present invention may contain 0.01 to 8% by weight of a composition containing bee venom extract as an active ingredient based on the total weight of the composition. When used in the above range, it is not only suitable for exhibiting the intended effect of the present invention but also can satisfy both the stability and safety of the composition and may be suitably used within the above range in terms of cost effectiveness. More specifically, when the content of the composition containing the bee venom extract as an active ingredient is lower than the above range, skin whitening or skin moisturizing effect due to the bee venom ingredient can not be sufficiently obtained, and when it exceeds the above range, The increase in effectiveness may not be significant.

The willow bark used in the willow bark extract is S. nigra, S. fragilis or S. alba, preferably Salixiniger (< RTI ID = 0.0 > S. nigra) or Salix alba (S. alba).

The extract of willow bark is a non-irritant substance when it is used directly as salicylic acid because it is a natural extract. The amount of Willow bark extract used in the present invention is preferably 0.001 to 10 wt%, more preferably 0.01 to 8 wt% , More preferably from 0.5 to 5% by weight.

The willow bark extract may be provided in the form of a solution, an extract or a powder by filtration or centrifugation, and the form thereof is not limited.

Preferably, the willow bark extract of the present invention can be prepared by the following process:

A first step of crushing the willow bark; The pulverized material pulverized in the above step 1 is purified water; Lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; Hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether and dichloromethane; Alternatively, they may be put into a mixed solvent, boiled in a reflux extractor equipped with a condenser for a certain period of time, extracted by heating, filtered, and then the residue is further extracted a predetermined number of times by the same method; The extracts are combined and filtered at room temperature to remove insoluble matter. The extract is concentrated in a distillation apparatus equipped with a cooling condenser at a reduced pressure of 50 to 80 ° C and then freeze-dried to obtain a dry extract of willow bark . In addition, the active ingredient may be extracted by immersion at room temperature for a considerable period of time.

The content of the willow extract contained in the composition according to the present invention is preferably 0.1 to 10% by weight based on the total weight of the composition. If the content is less than 0.1% by weight, %, The remarkable increase in the effect by the addition amount may not be exhibited.

In the present invention, "rice fermented extract" means a rice fermented product obtained by fermenting rice or a product resulting from the fermented product. Specifically, in the present invention, the term "rice fermentation extract" means rice fermented product itself, or the fermented rice may be obtained by a method such as extraction of a solvent (organic solvent such as water or ethanol), hot water extraction, cold extraction, reflux cooling extraction, It is meant to include extracted concentrates. The rice fermentation extract may be provided in the form of a solution, an extract or a powder by filtration or centrifugation, and there is no limitation on the form of the delivery.

Preferably, the rice fermentation extract of the present invention can be produced by the following production method:

1) mixing the rice with water and heating,

2) adding 0.05 to 10% by weight of yeast to the above-mentioned aged rice and saccharifying at 20 to 40 ° C for 10 to 30 hours,

 3) culturing at 15 to 40 DEG C for 1 to 10 days, and

4) Sterilization and concentration step.

Step 1) is a step of mixing rice with water and heat-grinding it. At this time, water is preferably added in an amount of 100 to 500% by weight, preferably 200 to 400% by weight, based on 100% by weight of the rice. It is also preferable to be called at 10 to 40 캜, preferably at 15 to 25 캜 for 1 to 24 hours, preferably for 2 to 6 hours. When the rice is cooked in water to make it contain proper moisture and then heat it to boil the rice, the starch particles of the rice gelatinize into the gel state and the micelle particles of the rice are formed to be wide, So that the fermentation can be promoted in the step of culturing in step 3). The heat aging step can be carried out for about 30 to 120 minutes, preferably for 40 to 60 minutes, with heat of 50 to 120 캜, preferably 70 to 90 캜, but the conditions can be suitably selected within a range obvious to a person skilled in the art .

Step 2) is a step of saccharification by adding nuruk to the above-mentioned gorgeous rice. This step is to saccharify the starch of rice to produce a monosaccharide, and to induce fermentation in the next step more easily by the monosaccharide. Nuruk can not only sacrifice the starch of rice but also prevent contamination of germs. In the present saccharification process, a composition containing a yeast mold can be used, and yeast can be usually used. Aspergillus kawachii, Aspergillus shirousamii, Aspergillus oryzae, Aspergillus niger, Aspergillus awamori, Aspergillus usamii, and Rhizopus sp. Can be used. But is not limited to.

In a specific example of the present invention, white koji was used as koji.

The koji is preferably added in an amount of 0.05 to 10% by weight, preferably 0.1 to 1% by weight, based on 100% by weight of the gorgeous rice. The koji is preferably 10 to 45 캜, After 10 to 48 hours, preferably 10 to 30 hours, at 40 ° C, various enzymes are leached to act on rice starch and glycosylation proceeds.

Step 3) is a step of culturing the saccharified rice to proceed fermentation. In this step, the saccharified rice can be cultured at 15 to 40 째 C, preferably at 15 to 35 째 C for 1 to 10 days, preferably 2 to 5 days. The micellar particles of rice are widely diffused by the above culturing step, and various enzymes can be smoothly performed, so that fermentation can be more surely proceeded.

Step 4) is a step of sterilizing and concentrating the fermented rice fermentation extract which has been fermented. The sterilization and concentration can be carried out according to a conventional method known in the art, and the sterilized and concentrated rice fermentation extract can be used as a food composition or a pharmaceutical composition by pulverization by spray drying or freeze drying.

The rice fermented extract of the present invention produced as described above is characterized by being free of fat (crude fat, saturated fat, and trans fat), and little or no cholesterol. Preferably, the rice fermented extract of the present invention may comprise the following nutritional composition:

0 to 10 wt% of water, 0 to 3 wt% of ash, 60 to 95 wt% of carbohydrate, 5 to 30 wt% of saccharides, 0 to 10 wt% of crude protein, 0 to 0.1 wt% of crude fat, 0 to 0.1 wt% More preferably 0.7 wt% of water, 1.1 wt% of ash, 83.4 wt% of carbohydrate, 20.7 wt% of saccharide, 0.1 wt% of trans fat, 0 to 0.1 mg / 100 g of cholesterol and 0 to 500 mg / 8.5 wt%, crude fat 0.0 wt%, saturated fat 0.0 wt%, trans fat 0.0 wt%, cholesterol 0mg / 100g, and sodium 47.08mg / 100g.

Preferably, the rice fermented extract of the present invention may contain 2 to 4% by weight of beta-glucan.

The method for preparing the mixture of the rice extract of the present invention and the fermented extract of rice comprises the steps of adjusting the rice to a moisture content of 5 wt% or less by drying, adjusting the rice to a moisture content of 5 wt% To 100 < 0 > C for 30 minutes to 1 hour to form a chemical condition so that the nutrient component of the raw material is eluted and the fermentation is facilitated. At the same time, Under anaerobic fermentation conditions, rice is fermented by solid fermentation using Bacillus subtilis such as Bacillus subtilis under dry cultivation condition, and then sterilization step is carried out to extract the extract with purified water. The extract is recovered and concentrated and purified. For further purification, it can be purified by ultrafiltration at 4 ° C. At this time, the separated extract is dried and the dry weight is measured to calculate the productivity.

The characteristics of the Bacillus sp. Strain used in the solid fermentation of the present invention are endogenous spore forming ability as Gram positive bacteria and have the form of enteric bacillus. It is also capable of growing under aerobic or flow anaerobic conditions but is not capable of growing under anaerobic conditions and has catalase production, casein, gelatin and starch resolution. As the growth conditions, it is possible to grow under 7% saline, and it is possible to grow at about 45 ° C, which is a fermentation condition, and the growth is active at about pH 5.0 to 7.5. It is also characterized in that an acid can be produced using glucose, arabinose, xylose and mannitol.

The cosmetic composition according to the present invention is characterized by containing 0.001 to 20% by weight, preferably 2% by weight, of the mixture of the above-prepared rice extract and the fermented extract of rice.

The rice raw material used in the extract of the present invention is characterized by being selected from the forms such as brown rice and white rice, and can be prepared by mixing with the solid fermentation extract of rice and subjecting to an ultrafiltration process to produce an extract containing fractions having excellent antioxidative effect have.

At this time, the mixture of rice extract and rice fermented extract according to the present invention shows an insufficient antioxidative effect and moisturizing elasticity function of the present invention when added in an amount of 0.001% by weight or less. When 20% by weight or more of the mixture is used, It is difficult to apply the final product due to intense smell and color due to flavor.

The cosmetic composition having an anti-aging effect of the skin according to the present invention may have conventional formulations and may be, for example, a softening agent, a facial essence (mask), a nutrition lotion, a nutritional cream, a sunblock cream, a body lotion, ≪ / RTI > The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art. Hereinafter, a representative nutritional cream and a nutritional lotion composition of a cosmetic composition having an anti-aging effect will be described as an example.

In addition, the extract of the present invention having excellent anti-aging effect on the skin can be used according to a conventional method of use, and the use frequency of the extract may be varied depending on the skin condition or taste of the user. In addition, it can be applied to hair products such as soap, facial foam, body cleanser, cleanser, shampoo, rinse, etc., and patch form such as mask pack is also possible. In addition, if necessary, it can be blended with components generally used in external preparations for skin such as an aqueous component, an oily component, a surfactant, a moisturizing agent, a thickening agent, an antiseptic, a perfume pigment and the like.

Examples of the surfactant used in the composition of the present invention include lecithin, xanthan gum, cetyl alcohol, and the like. Lecithin is a naturally occurring diglyceride mixture of stearic acid, palmitic acid and oleic acid, which is bound to the choline ester of phosphoric acid and can be extracted mostly from eggs or beans and used as a skin conditioning agent, skin softening agent, surfactant - Used as an emulsifier.

It is a high molecular weight heteropolysaccharide gum obtained by fermentation of carbohydrate by pure cultivation of black beans with black soybean flour. It mainly extracts from polysaccharide of plant and increases viscidity. It enhances skin elasticity, Active agents - emulsifiers, thickening agents - water soluble. Cetyl alcohol is alcohol extracted from coconut and acts as a lubricant. It mixes well the water and oil components contained in cosmetics. It acts as emulsifying stabilizer and viscosity controlling agent. It can make highly viscous cosmetics and increase the moisture of cosmetics. Smoothes skin and supplies oil.

In the composition of the present invention, glycerin, hyaluronic acid, butylene glycol, and allantoin may be used as the moisturizing agent. Glycerin is a complex solvent and has the function of fixing it as an active ingredient, which smoothes the skin and has effects such as fragrance, skin moisturizer, skin care agent and viscosity reducing agent. It is stable and has excellent moisturizing effect and is mainly used in cosmetics. Hyaluronic acid is a combination of protein and polysaccharide. It is also called glycosaminoglycan. It is bonded to connective tissue fiber-collagen and elastin of the dermis, filling the space between them. Since it is a polymer compound, its molecular size is very large. It is a natural moisturizing agent that acts as a polymer of water-soluble substances and strongly catches moisture on the epidermis. It is present in many human skin and earthworms and acts as a skin conditioning agent, thickening agent and moisturizer. Butyleneglycol is a large organic alcohol that is used as a powerful moisturizing agent to attract moisture to the skin. Allantoin is a naturally occurring substance in nitrogen fertilizer, which not only functions to moisturize the skin, but also heals wounds, manages ulcers, and improves smooth and rough skin.

In the composition of the present invention, natural preservative GSE (Grapefruit Seed Extract) or MSE (Multi-Scutellaria etracts) can be used as a preservative. GSE is a grapefruit seed extract. MSE is extracted from the roots of Lamiaceae plants. It has antimicrobial effect on inflammation and skin troubles of various skin, antioxidant effect, whitening effect, anti-aging effect.

In another aspect, the composition for external application for skin according to the present invention may be manufactured in the form of a cream, a lotion, an emulsifier, a liniment, etc., which are semi-solid preparations, but not limited thereto, and may be a suitable colorant, a flavoring agent, A surfactant, and the like. In addition, the composition for external application for skin according to the present invention may be prepared as a liquid preparation, and may be a solution, a suspension, or an emulsion such as water, an alcohol, a propylene glycol solution or the like, but is not limited thereto. Suitable colorants, , A viscosifying agent, and the like.

Specifically, the external preparation for skin may be a pharmaceutically acceptable base such as vaseline or stearyl alcohol; Suitable pharmaceutically acceptable surfactants such as polysorbate, sorbitan sesquioleate, and the like; A suitable pharmaceutically acceptable humectant such as glycerin; A suitable pharmaceutically acceptable solvent; And a conventional external preparation for skin preparation in which a flavoring agent, a coloring agent, a stabilizer, a tackifier and the like are homogeneously mixed.

The present invention relates to an anti-aging cosmetic composition containing bee venom extract, willow bark extract and rice fermented extract as an active ingredient. The cosmetic composition for anti-aging not only has an antioxidative effect, an effect of biosynthesis of collagen fibers, and an effect of inhibiting the activity of hyaluronase. The skin elasticity enhancement effect and skin safety are excellent.

Hereinafter, the present invention will be described in detail. However, the following examples, reference examples and experimental examples are illustrative of the present invention, and the present invention is not limited thereto.

Production Example 1. Production of bee venom and preparation of bee venom extract

Bee venom was obtained by electric shock method. The bee venom obtained from the electric shock method was poured with distilled water to float bees or debris in bee venom. Filtered suspended bees or debris to obtain purified beacon.

The obtained bee venom was extracted with 70% ethanol. Specifically, 70% ethanol corresponding to 3 times the volume of the bee venom obtained was added, and the mixture was extracted and degreased with stirring at room temperature five times. 70% ethanol was added to the degreased bees poison four times, and the mixture was refluxed three times and then immersed at 15 ° C for 2 days. The precipitate was filtered and centrifuged to separate the residue and filtrate, and the separated filtrate was concentrated under reduced pressure to obtain an extract. After the resulting extract was suspended in water, the pigment was removed using ether, and the aqueous layer was extracted with butanol five times. The obtained organic solvent layer was concentrated under reduced pressure to give an extract. The resulting extract was dissolved in methanol, ethyl acetate was added, and the resulting precipitate was dried to obtain a bee venom extract.

Production Example 2. Preparation of Birch Bark Extract

100 g of milled pulverized product obtained by purchasing commercially available willow bark (S. Nigra Bark) was added to 3 L of purified water for solvent, and the mixture was boiled for 3 hours in a reflux condenser equipped with a condenser and then filtered through 100 mesh filter cloth. . The extracts were combined and filtered twice with Whatman filter paper at room temperature to remove insoluble matter. The extracts were concentrated under reduced pressure at 60 ° C in a distillation apparatus equipped with a cooling condenser, and then lyophilized to obtain a dry extract of willow bark.

Production Example 3. Preparation of rice fermented extract

The rice was cooked at 15 ~ 25 ℃ for 2 ~ 6 hours. The rice was cooked at 80 ℃ using 6 times water. The temperature was lowered to 30 ℃ and the rice flour was added to the rice at 10%. Next, saccharification was carried out at 30 DEG C for 12 hours, followed by culturing at 25 DEG C for 48 hours, followed by sterilization and filtration to obtain powder (spray drying or lyophilization). Beta-glucan was selected as an index material for the rice fermentation extract prepared above, and analyzed according to the "Health Functional Food Slot II.3.4.3 Beta Glucan ". As a result, beta-glucan was found in the fermented rice extract ~ 4%, and exactly 3%.

In addition, it was confirmed that the nutritional components contained in the fermented rice extract were as follows:

A trans fat of 0.0% by weight, a cholesterol of 0 mg / 100 g, and a sodium of 47.08% by weight, water content of 7.0% by weight, ash content of 1.1% by weight, carbohydrate of 83.4% by weight, saccharides of 20.7% by weight, crude protein of 8.5% by weight, crude fat of 0.0% mg / 100g, calories 368kcal.

Production Example 4. Preparation of complex extract

The bee venom extract, willow bark extract and rice fermentation extract prepared in Preparation Examples 1 to 3 were mixed at a ratio of 0.5: 0.5: 1, respectively, and used in the following Production Examples and Experimental Examples.

Production Example 5. Preparation of nutritional cream

The nutritional cream was prepared using the compound extract prepared in Preparation Example 4, and the total weight of bee venom, willow bark and rice fermentation extract was 8.00% by weight based on the total weight of nutritional cream.

First, purified water, a complex extract, Carbomer, Disodium EDTA, Butylene Glycol and Methyl Paraben were weighed according to the composition ratios shown in Table 1 below, And mixed at 2 DEG C to complete a water phase. In addition, according to the composition ratio shown in Table 1 below, it is possible to use propyl paraben, Macademia nut oil, mineral oil, dimethicon, caprylic caprylic, Capric Triglyceride) was weighed and mixed at 80 ± 2 ℃ to complete the oil phase.

Next, the oil phase was added to the water phase so that the two phases were mixed with each other to prepare an emulsion-type mixture. Triethanolamine, a neutralizing agent, was added to the mixture, and the mixture was cooled to 45 캜. Then, flavorings (Argeville Co., M.A. CO 25106B) were added and the mixture was cooled to 35 캜 to prepare nutritional cream.

The control nutrient cream was prepared according to the composition ratio shown in Table 1 below, except that bee venom extract, willow bark extract, and rice fermentation extract were not used as a control group and the weight ratio of purified water was 84.95%.

division Raw material name Nourishing cream containing complex extract Control Nutrition Cream Awards One Purified water 76.95 84.95 2 Complex extract 8.00 - 3 Carbomer 0.15 0.15 4 Disodium edetate 0.05 0.05 5 Butylene glycol 4.00 4.00 6 Methyl paraben 0.20 0.20 Paid 7 Propylparaben 0.10 0.10 8 Macadamia nut oil 4.00 4.00 9 Mineral oil 3.00 3.00 10 Dimethicone 1.00 1.00 11 Caprylic Capric Triglyceride 2.00 2.00 corrector 12 Trylethanolamine 0.25 0.25 additive 13 Spices 0.30 0.30

Production Example 6. Production of Nutrition Lotion

In Preparation Example 4, the nutritional lotion was prepared using the complex extract, and the extract was contained in an amount of 8.00 wt% based on the total weight of the nutritional lotion.

First, the purified water, bee venom extract, willow bark extract, rice fermented extract, carbomer, disodium edetate, butylene glycol, and methylparaben were weighed and mixed at 80 ± 2 ° C according to the composition ratio shown in Table 2 below. (water phase). Further, according to the composition ratio shown in the following Table 3, propylparaben, macadamia nut oil, mineral oil, dimethicone, sorbitan sesquioleate and polysorbate 60 were weighed, And mixed at 2 캜 to complete an oil phase.

Next, the oil phase was added to the water phase so that the two phases were mixed with each other to prepare an emulsion-type mixture. Triethanolamine, a neutralizing agent, was added to the mixture, cooled to 45 ° C, perfumed (Argeville, M.A. CO 25106B) was added and the mixture was cooled to 35 ° C to prepare a lotion.

A control lotion was prepared according to the composition ratio shown in Table 2 below, with the weight ratio of purified water being 86.13%, without using the complex extract as a control.

division Raw material name Nutritional lotion containing complex extract Control Nutrition Lotion Awards One Purified water 78.13 86.13 2 Complex extract 8.00 - 3 Carbomer 0.15 0.15 4 Disodium edetate 0.20 0.20 5 Butylene glycol 1.00 1.00 6 Methyl paraben 0.20 0.20 Paid 7 Propylparaben 0.10 0.10 8 Macadamia nut oil 2.00 2.00 9 Mineral oil 6.00 6.00 10 Dimethicone 2.00 2.00 11 Sorbitan sesquioleate 0.40 0.40 12 Polysorbate 60 1.60 1.60 corrector 13 Trylethanolamine 0.15 0.15 additive 14 Spices 0.25 0.25

Experimental Example 1. Antioxidative Effect of Compound Extract

To confirm that the complex extract prepared in Preparation Example 4 had an antioxidative effect, a thiobutylacetate (TBA) test was conducted using a lipid peroxidation system by Fenton's reaction.

First, 25 mM Trizma hydrochloric acid / 0.75 mM potassium chloride buffer solution was prepared by adding 0.2% sodium dodecylsulfoxide (SDS), 10 mol of ferrous chloride and 2 mol of hydrogen peroxide to 10 mL of ethyl linoleate.

Next, 0.01 g of the complex extract of Preparation Example 4 was dissolved in 0.1 mL of dimethylsulfoxide (DMSO) solution. The solution in which the complex extract was dissolved and 5 mL of 25 mM Trizma hydrochloric acid / 0.75 mM potassium chloride buffer solution were mixed and cultured at 55 ° C for 16 hours, followed by addition of 50 mL of 4% butylhydroxyltoluene , The reaction was stopped and then placed in a 0.3 mL test tube. 3.0 mL of 0.05 M hydrochloric acid and 4.0 mL of butanol were added to the 0.3 mL test tube, and the absorbance of the sample was measured at 535 nm by centrifugation.

The absorbance of the sample was measured at 535 nm as a control. The measurement results of the absorbance are shown in Table 3 below.

Manufacturing example Absorbance The complex extract of Preparation Example 4 0.311 Control group 0.075

Using the absorbance in Table 3, the antioxidant effect was calculated. The antioxidant effect can be calculated by the following formula, and the results are shown in Table 4 below.

              Test group absorbance - Control absorbance

Antioxidant effect = ------------------------------ X 100

Control absorbance

sample Antioxidative effect(%) The complex extract of Preparation Example 4 89.25 Vitamin E 45.20

As can be seen from the results of Table 4 above, the antioxidative effect of the complex extract of Preparation Example 4 (present invention) obtained by using an extraction solvent prepared by mixing ethanol: butylene glycol: distilled water at a weight ratio of 2: 1: Was 89.25, while the antioxidant effect of vitamin E, which is widely known to have antioxidant effect, was only 45.20. From the above results, it can be seen that the antioxidative effect of the complex extract of the present invention is very high.

Experimental Example 2. Effect of Combined Extracts on Collagen Fiber Biosynthesis

A test was conducted to confirm that the complex extract prepared in Preparation Example 4 had an effect of biosynthesizing collagen fibers.

First, human fibroblast (ATCC-28) was put into a culture flask and cultured until it grew to about 70 to 80%. The cultured fibroblasts were treated with a complex extract, and the complex extracts were treated at a ratio of three kinds of extracts: 0.01 wt%, 0.10 wt%, and 1.00 wt%, based on the total weight of the fibroblasts. After that, the ratio (%) of collagen fiber biosynthesis of the fibroblasts treated with the complex extract was measured using a commercially available kit for measuring collagen fiber biosynthesis (BCA protein assay kit, pierce 23225).

Then, the untreated fibroblasts were used as a control group. In the case of not irradiating ultraviolet light, control group 1 was used, and when ultraviolet light was irradiated, control group 2 was used. After the above steps, the fibroblast collagen fiber biosynthesis rate %) Was measured.

The percentages of collagen fiber biosynthesis (%) as measured above are shown in Table 5 below.

sample Collagen fiber biosynthesis rate (%) Compound extract 0.01 (%) Compound extract 0.10 (%) Compound extract 1.00 (%) Fibroblasts treated with complex extracts 176 184 197 Control 1 96 96 96 Control group 2 68 68 68

As can be seen from the results of Table 5, the collagen of fibroblasts treated with the complex extract of Preparation Example 4 obtained using an extraction solvent prepared by mixing ethanol: butylene glycol: distilled water at a weight ratio of 2: 1: The percentage of fiber biosynthesis (%) showed about twice the collagen fiber biosynthesis rate (%) compared with the control 1 or control 2 not treated with the combined extract.

As can be seen from the above, the fact that the fibroblasts treated with the complex extract of the present invention have a high rate of collagen fiber biosynthesis shows that the complex extract of the present invention has an excellent effect in preventing wrinkles caused by aging, will be.

Experimental Example 3. Inhibitory Effect of Compound Extract on Hyaluronase Activity

A test was conducted to confirm that the combined extract prepared in Preparation Example 4 had an effect of inhibiting the activity of hyaluronase (Sigma, type 4). The hyaluronase is an enzyme that degrades hyaluronic acid involved in moisture retention of cells. When the activity of hyaluronase is inhibited, the stability of hyaluronic acid is enhanced, so that the ability of the cells to retain moisture can be enhanced.

First, 10 mg of the combined extract was dissolved in 100 mL of 0.1 M acetic acid buffer solution (pH 4.0) and placed in a test tube. Thereafter, 50 mL of acetic acid in which 0.1 mg of hyaluronase was dissolved was added to the test tube, and the mixture was allowed to stand at 37 캜 for 20 minutes.

Next, 250 mL of acetic acid buffer solution in which 200 mg of sodium hyaluronate had been dissolved was added to the test tube, and the mixture was allowed to stand at 37 DEG C for 40 minutes. Thereafter, 100 mL of 0.4 M sodium hydroxide was added to the test tube, and the mixture was cooled.

Next, 100 mL of a boronic acid buffer solution (pH 9.0) was added to the test tube, and the sample was incubated at 37 ° C for 20 minutes. The absorbance was measured at 585 nm.

The absorbance of the untreated sample was measured at 585 nm as a control. The measurement results of the absorbance are shown in Table 6 below.

sample Absorbance Complex extract 0.198 Control group 0.102

Using the absorbance in Table 6, the inhibitory effect of hyaluronase was calculated. The activity inhibitory effect of hyaluronase can be calculated by the following equation, and the results are shown in Table 7 below.

                                Test group absorbance - Control absorbance

Inhibition of hyaluronase activity = ------------------------------ X 100

                                  Control absorbance

sample Inhibitory Effect of Hyaluronase (%) Complex extract 92.77

As can be seen from the results of Table 7 above, the activity of inhibiting the activity of hyaluronase of the complex extract of Preparation Example obtained using an extraction solvent prepared by mixing ethanol: butylene glycol: distilled water at a weight ratio of 2: 1: 1 was 92.77 %. From the above results, it can be seen that the compound extract of the present invention has a very high effect of inhibiting the activity of hyaluronase.

Experimental Example 4. Skin Elasticity Enhancement Effect of Nourishing Cream

The nutritional cream prepared in Preparation Example 5 and the control nutrition cream were tested to see if they had the effect of improving skin elasticity.

For this study, 35 healthy women aged 28 to 40 years (mean age 34 years) were divided into two groups. Nutritive cream and control nutrition cream were added to each group for 12 weeks 2 times a day.

Skin elasticity of the nutrient cream before use (0 weeks) and skin elasticity after using the nutritional cream for 12 weeks were measured with a skin elasticity meter (Cutometer SEM575, Courage + Khazaka). The net elasticity (R5) can be calculated using the measured values, and the closer to 1 the sealant number, the better the skin elasticity enhancement effect. The physical modulus of elasticity was calculated by the following formula, and the results are shown in Table 8.

Actual Young's modulus (R5) = R5 (12 weeks) - R5 (0 weeks)

The actual modulus of elasticity (R5) Nourishing cream containing complex extracts 0.19 Control Nutrition Cream 0.015

As can be seen from the results of Table 8, the real elasticity coefficient when the nutritional cream containing the compound extract of the present invention (the present invention) was used was 0.19. And 1267% higher than the control nutrition cream containing no complex extract. In conclusion, it was found that the nutritional cream containing the complex extract of the present invention effectively promoted skin elasticity.

Experimental Example 5. Cytotoxicity Experiment

The cytotoxicity of the compound extract prepared in Example 4 was measured by using the mouse-derived dermal fibroblast according to the following method.

The mouse skin fibroblasts of the logarithmic growth phase were inoculated into a 96-well plate at 10,000 per well and cultured in DMEM medium (80 μl) for 24 hours. The final concentration of the complex extract was 0, 50, 100, 200 and 500 g / , 15 μl of a 5 mg / ml MTT solution was added thereto. After 4 hours, a dimethylsulfoxide solution was added at a rate of 150 μl per well And the absorbance was measured at 570 nm using a microplate reader.

The cytotoxicity of the combined extracts was expressed by the cell viability (%) and was calculated by the following equation.

                  Absorbance at the time of extract treatment

Cell survival rate (%) = ---------------------- X 100

Control absorbance

Treatment concentration (ug / ml) of compound extract Cell viability (%) 0 100 50 102.4 100 100.7 200 104.2 500 101.3

As shown in Table 9, the complex extract did not show significant cytotoxicity as compared with the negative control in itself.

Experimental Example 6: First-order irritation test on human body (closed pellet)

In order to examine the safety of the combined extracts, a first stimulation test was performed on human body through a human patch test as described below.

Twenty healthy men and women were enrolled in the CTFA Guidelines (The Cosmetic, Toiletry and Fragrance Association, Inc., Washington, DC, 20036, 1991). 15 mg / 15 μl of the sample and solution was dropped into a Finn chamber, and this was placed on the arm of the human body to be tested and fixed with a tape. Subsequently, the skin was reacted for 24 hours, and then the patch was removed. After 2 hours, the skin reaction of the test site was judged according to the following criteria. Table 10 below shows the criterion of judgment, and Table 11 shows the skin irritation index of each sample.

reaction score Symptoms and judging criteria - 0 No response +/- 0.5 Weak erythema + One Clear erythema ++ 2 Severe erythema with edema +++ 3 Severe erythema, blisters, edema

Skin irritation index = [Σ (number of points × number of stimuli)] / total number of test subjects × number of total stimulation evaluations

sample Number of subjects Skin irritation index Bee venom extract 1% 30 0.0 Willow bark extract 2% 30 0.0 Rice fermented extract 2% 30 0.0 Complex extract 30 0.0

As shown in Table 11, the combined extract of bee venom extract, rice bran extract, rice fermentation extract, bee venom, willow bark and rice fermentation extract, which are effective ingredients of the cosmetic composition according to the present invention, ≪ / RTI >

Claims (8)

An anti-aging cosmetic composition comprising bee venom extract, willow bark extract and rice fermented extract as active ingredients
The antioxidant cosmetic composition according to claim 1, which comprises 0.0001 to 30% by weight of a composition containing the bee venom extract as an active ingredient.
The antioxidant cosmetic composition according to claim 1, wherein the bee venom extract is obtained by an electric shock method.
The method according to claim 1, characterized in that the willow bark used in the willow bark extract is S. nigra, S. fragilis or S. alba By weight based on the total weight of the cosmetic composition.
The cosmetic composition for preventing aging according to claim 1, wherein the fermented rice extract is a fermented rice obtained by fermenting rice or a product produced from the fermented product.
The method of claim 1, wherein the willow bark extract comprises: a first step of grinding a willow bark; The pulverized material pulverized in the above step 1 is purified water; Lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; Hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether and dichloromethane; Alternatively, they may be put into a mixed solvent, boiled in a reflux extractor equipped with a condenser for a certain period of time, extracted by heating, filtered, and then the residue is further extracted a predetermined number of times by the same method; A third step of removing the insoluble matter by filtration at room temperature after the addition of each of the above extracts, followed by concentration under reduced pressure at 50 ° C to 80 ° C in a distillation apparatus equipped with a cooling condenser, followed by lyophilization.
[3] The method of claim 1, wherein the rice fermented extract comprises: 1) mixing rice with water and heating and boiling; 2) adding 0.05 to 10% by weight of yeast to the grated rice and saccharifying at 20 to 40 ° C for 10 to 30 hours; 3) culturing at 15 to 40 DEG C for 1 to 10 days, and 4) sterilizing and concentration.
The method according to claim 7, wherein the yeast is selected from the group consisting of Aspergillus kawachii, Aspergillus shirousamii, Aspergillus oryzae, Aspergillus niger, Aspergillus awamori, Aspergillus usamii and the like or Rhizopus sp. Wherein the cosmetic composition is at least one kind selected from the group consisting of cosmetics and cosmetics.