KR20170133721A - A composition comprising extract of inula japonica thunberg, epimedium koreanum and knotgrass as active ingrediant for functional cosmetics - Google Patents

Abstract

The present invention relates to a method for producing extracts of Bacillus subtilis, Bacillus thuringiensis L. and Mamacillus complexes having synergistic effect by mixing among extracts, and a functional cosmetic composition prepared therefrom, outstanding. Particularly, the natural extracts of the present invention are excellent in antioxidation, cell protection, skin moisturizing and skin soothing effects, and the cosmetic compositions containing the natural plant extract as an active ingredient are excellent in antioxidation, skin moisturizing And cosmetics for skin soothing.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a cosmetic composition, a method for preparing the same, and a functional cosmetic composition prepared from the same. BACKGROUND ART [0002]

The present invention relates to a method for producing extracts of Bacillus subtilis, Bacillus thuringiensis L. and Mamacillus complexes having synergistic effect by mixing among extracts, and a functional cosmetic composition prepared therefrom, outstanding.

Human is experiencing the aging phenomenon that physiological function declines with time, and skin is one of the index that can judge the degree of biological aging. Since the skin is always exposed to the external environment, a lot of skin changes due to aging can be attributed to the influence of external factors. In particular, the inflammatory response to external factors, active oxygen, and ultraviolet light are considered to be the main causes.

Active oxygen causes damage to skin cells and tissues, destroying the antioxidant defense system, and is involved in skin aging as well as biological aging. It causes functional damage to various organs, thereby inducing various diseases and promoting aging. In addition, due to persistent oxidative stress, it causes damage of bio-components such as activation of proteolytic enzymes, collagen and elastin cleavage, and hyaluronic acid cleavage, and inhibits skin inflammation and skin immune function, thereby increasing bacterial infection or cancer. Therefore, skin cells should be protected by inhibiting active oxygen production, inhibiting inflammation caused by active oxygen, and increasing skin moisturizing power.

The human body contains about 72% moisture, which keeps the skin moist and protects it gently. Therefore, moisture retention in the skin is a very important factor for maintaining a healthy skin, and it may be the most basic factor that can cope with weakening of the skin resistance over time. Moisture is maintained in the stratum corneum to maintain the elasticity and flexibility of the skin. Normal skin contains about 30% water and water-soluble ingredients. The skin is largely divided into three layers: epidermis, dermis, and subcutaneous tissue in order from the outside. It has a function to protect the internal organs of the body from changes in temperature and humidity, ultraviolet rays, and other physical and chemical external stimuli. In particular, the epidermis plays an important role in preventing water evaporation inside the human body.

The epidermis is divided into the stratum corneum, granular layer, the superficial layer and the basal layer in order from the outside, and the cells of the stratum corneum act like bricks and the intercellular lipids between the keratinocytes function as the mortar to constitute the skin barrier. (J. Invest. Dermatol 80 (Suppl.), 44-49, 1983). In addition, a healthy human keratinocyte has a high concentration of natural moisturizing factor (NMF) to help maintain moisture in the skin. For example, substances such as amino acids are water-soluble, Thereby suppressing drying. (J. Invest. Dermatol., 54, 24-31, 1970).

There are two major types of water in the stratum corneum: bound water and free water. It is reported that the bound water is bound in molecular form with the ions, amino acids and proteins in the stratum corneum, and it is reported that water which is usually present in the dried stratum corneum, which is dried and contained in the stratum corneum, is about 5% have. In addition, water molecules that are in a form that binds very rapidly to the stratum corneum but does not fall off even in a dry state and is in a relatively strongly bound state is called a secondary binding water. Therefore, when moisture is bound to the stratum corneum and the water exists in the form of bonded water, particularly in the form of secondary bonded water, the moisture is bound to the stratum corneum, and the skin is moisturized even in a dry state with low external humidity.

However, as it is nowadays, there is a tendency to adjust the temperature of the air / heating due to the change of environment or life pattern, the stress caused by various stresses and environmental pollution in the social life, frequent washing according to make- Due to various causes such as aging of the skin, the moisture of the stratum corneum is decreased, and the skin becomes dry, the surface becomes rough, the skin becomes loose, the moisture is lost and the skin looks livelier. .

Conventionally, a method of increasing the moisture retention in the stratum corneum has been performed by using a humectant having a property of absorbing moisture as a humectant and an occlusive moisturizer preventing evaporation of water. As humectant, there are substances such as glycerin, propylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, 2-pyrrolidone-5-carboxylate and the like, (J. Invest. Dermatol. (5), 731-740, 1994). However, the stability of the emulsified formulations is maintained even though the lipid components such as ceramides and essential fatty acids and lipid complexes have been used as the closed moisturizer And there are disadvantages in manufacturing transparent gel-like products.

Therefore, there is a need for a simple and immediate composition that can minimize adverse effects on the skin due to stress, that is, to quickly normalize skin barrier protection and keratin turnover, and to rest on the skin, have.

Jinbukcho is a perennial herb that grows in the mountains and fields of each country. It grows well in the places where the mountains and plants are drenched. It is semi-shrub or sunny plant, 20-60㎝ tall, 5-10㎝ long, 1 ~ 3㎝ wide, slanting, long oval, with sparse rim. The flower is yellow and hangs on the tip of the branch and the end of the stem. The diameter is about 3㎝, and the fruit runs around October and has a hair of about 0.5㎝ long. Unlike other chrysanthemums, flowers are characterized by narrow petals and long leaves. Young lilies are used for edible purposes, and flowers are used for medicinal purposes.

Inula japonica Thunberg contains Sesquiterpenoid lactone, britanin and inulin on the ground and Quercetin as a flower embankment, It contains a large amount of sterol components such as isoquercetin, caffeic acid, cholorogenic acid, inulin, and taraxasterol.

It is preferable that the bamboo shoot extract is extracted from Bongbokcho flowers. This is because it enhances the body's immune system, and the antioxidant, anti-inflammatory and antiallergic Flavonoid components are higher than other parts.

Korean Patent Publication No. 2011-0087448 entitled " Composition for preventing and treating inflammation or allergic diseases containing the compound isolated from the prey-laden extract as an active ingredient "includes " a compound isolated from the lyophilized Inhibit the production of cyclooxygenase-2 (COX-2) dependent prostaglandin and the production of leukotriene from mast cells that cause allergic diseases and inhibit the release of histamine from mast cells And thus it can be usefully used as a medicament or a health functional food for the prevention and treatment of various inflammatory or allergic diseases.

J. Soc. Cosmet. Scientists Korea Vol. 35, No. 3, September 2009, 209-217, "Antimicrobial and Antioxidant Activity of Flower Extracts from Bamboo Vulgid Flower" has the antimicrobial activity against the skin topical bacterium, the antioxidant activity and the tyrosinase inhibitory activity, Quot; has been disclosed.

Epimedium koreanum is a perennial plant of the perennial herbaceous perennial plant, which is a plant native to Korea and northeastern China. It grows mainly in the shade of the mountain in the mountainous region, and the rootstock extends to the side and has many roots. Stem is united, height is 30cm, thin, hairless, and the bottom is surrounded by scaly leaves. The upper part of the stem is divided into 3 branches and 3 leaves per branch. The leaves from the root are blunt and the petiole is long. Leaf is 5 ~ 13.5cm long ovate with pointed end, lower part is heart-shaped and has fringed sawtooth on edge. The flower blooms in May and runs on the bottom of the stem, forming a gunshot at the end of the stem. The fruit is cornelian and has a cylindrical shape with a pointed end with a length of 10-13 mm. In one room, the whole plant is used as a medicinal substance called yin yang (阴羊羊 藿), which has an aphrodisiac, tang, gangjeong, and a strong wind effect. In the private sector, it is used for negative (negative), nervous breakdown, forgetfulness, hysteria, and lack of erection.

It is a perennial plant belonging to the genus Porphyra spp., And has no hair on the whole. The stem is divided and spreads on the top, there are many vertical lines, and the nodes are swollen. The height is about 10 to 40 cm, and the leaves are short oval, long oval, 2 to 4 cm long, with no hairs, and the back side has a middle vein. The sheath is membranous, irregularly torn, and has a thin vein. It blooms on small green flowers on the leaf axil in July-September. There are 5 calyxes, oval, greenish white or pinkish red, 6 ~ 8 stamens, 3 styles, fruit is abundant in water, sparsely dispersed, and glossy. Flowering period is from June to July, young leaves are edible as herbs, and outposts are used as medicinal plants. It is widely used throughout the world and has a variety of efficacy, so it is being used as a private herb in many countries. In particular, it has been known that there is diuretic action, blood pressure lowering action, accelerated action of pancreatic activity and contraction of uterus, and also there is pulmonary tuberculosis, neuroleptic, antiseptic, nephrolithiasis, gastroenteritis, child diarrhea, respiratory disease, gastric ulcer and headache It is also known to be used in many other diseases.

However, none of the above documents disclose the synergistic effect of the active ingredient when the natural plant extracts are mixed. Accordingly, the present inventors have studied the activity of the extracts of Bombyx mori, Sangjiwai lobster and Mamallipu complex and their activities. When the components are independently contained in the cosmetic composition, they are more excellent in antioxidation, cell protection and skin moisturizing , And skin soothing effect.

KR publication 10-2011-0087448 (August 3, 2011)

 J. Soc. Cosmet. Scientists Korea Vol. 35, No. 3, September 2009, 209-217

The present invention provides a method for producing extracts of Bombyx mori, Bacillus thuringiensis and Mamacil complexes and a functional cosmetic composition containing the same, wherein the skin-related effect is further enhanced by the synergistic effect, as compared with the cosmetic composition containing the extract of the present invention .

The present invention has been conceived in order to obtain a complex extract having a better activity enhancing effect than the prior art,

The present invention provides a cosmetic composition comprising Aspergillus oryzae L., Mulberry Leaf Lobster, and Madillip composite extract as active ingredients.

In addition, the cosmetic composition of the present invention comprises the combined extracts obtained by mixing the fragrance of the genus Prunus mume, the three leaves of Leek mung bean and the mallow of the present invention in an amount of 1 to 15: 1 to 15: 1 to 15 by weight.

In addition, the cosmetic composition of the present invention provides a cosmetic composition characterized by an antioxidative activity effect.

In addition, the cosmetic composition of the present invention provides a cosmetic composition characterized by having a cytoprotective effect.

Further, in the present invention, the cosmetic composition is provided with a skin moisturizing effect.

In addition, the cosmetic composition of the present invention provides a cosmetic composition characterized by a skin soothing effect.

In addition, the cosmetic composition of the present invention is characterized by being a hot water extract or a solvent extract.

In the present invention, the solvent may be at least one selected from the group consisting of water, fermented alcohol, ethanol, methanol, butanol, propanol, ethyl acetate, isopentyldiol, glycerin, ethylene glycol, propylene glycol, butylene glycol, chloroform, dichloromethane, , At least one solvent selected from the group consisting of acetonitrile, petroleum ether and diethyl ether.

The cosmetic composition according to the present invention may further contain 0.001 to 10% by weight of the extract of Aspergillus oryzae.

In the present invention, the cosmetic composition for antioxidation, cell protection, skin moisturizing and skin cleansing may be at least one selected from the group consisting of lotion, cream, essence, cleansing foam, cleansing water, pack, patch, body wash, body lotion, body oil, , A hair conditioner, a hair gel, a skin adhesive gel, a foundation, a lipstick, a mascara, and a makeup base product.

Also, there is provided a method for manufacturing a bean curd, comprising:

Heating the mixed natural plant at 80 to 120 캜 to obtain an extract;

Filtering the extract with a filter to obtain a filtrate; And

And then concentrating and lyophilizing the filtrate to obtain a combined extract.

The cosmetic composition containing the natural plant extract as an active ingredient is excellent in antioxidation, cell protection, skin protection, skin moisturizing effect, and skin soothing effect, It can be useful as a cosmetic for moisturizing and soothing the skin.

Hereinafter, the present invention will be described in detail.

As used herein, the term "Sanko Rhizome extracts" means those obtained by extracting from various organs or parts (e.g., fruits, leaves, flowers, roots, stems, branches and shells) of Sanko leaves, ≪ / RTI > of the whole body of the plant.

According to one embodiment of the present invention, firstly, the granulomatosus sphaeroa, the sage lily of the valley and the falciparum are cut out and mixed. Then, a solvent is added to the natural mixture to prepare a combined extract. Alternatively, the complex extract of Aspergillus oryzae may be used in the form of a combined extract obtained by extracting each of the components with a solvent.

According to an aspect of the present invention,

The mixture is extracted by mixing at a weight ratio of 1: 15: 1 to 15: 1 to 15, and the mixture is extracted by mixing at a weight ratio of 1: 10: 1 to 10: 1 to 10: desirable. Particularly, it is most preferable to mix and extract Gwangbukcho: Sajjiwolbyeol: mallowwool = 1 ~ 5: 1 ~ 5: 1 ~ 5 weight ratio.

The combination extract shows excellent antioxidation, cell protection, skin moisturizing and skin soothing effects. A more detailed understanding of this may be made through the following examples and experimental examples.

The combined extracts of Ganpungcho, Sangjiwoobyphi and Mamallifolia are hot water extracts, water, fermented alcohol, ethanol, methanol, butanol, propanol, ethyl acetate, isopentyldiol, glycerin, ethylene glycol, propylene glycol, butylene glycol, chloroform, dichloro Is a solvent extract by at least one solvent or at least one solvent selected from the group consisting of methane, hexane, acetone, acetonitrile, petroleum ether and diethyl ether, and is preferably a hot water extract.

The complex extract preferably contains 0.001 to 20% by weight, and preferably 0.01 to 10% by weight based on the total weight of the composition. Further, it is most preferable to contain 1 to 5% by weight. If the weight ratio is less than 0.001% by weight, a desired effect can not be expected. If the weight ratio is more than 20% by weight, the increase of the effect is insignificant, cytotoxicity may be caused, and formulation stability may be deteriorated.

Each of the above extracts has antioxidant, cell protection, skin moisturizing effect and skin soothing effect as well as an increased effect due to the combination of each component. A more detailed understanding of this may be made through the following examples and experimental examples.

The cosmetic composition of the present invention may be manufactured into various cosmetic composition formulations and may be in the form of, for example and without limitation, lotions, creams, essences, cleansing foams, cleansing waters, packs, patches, body wash, body lotions, A cream, a cream, a paste, a gel, a shampoo, a rinse, a hair conditioner, a hair gel, a skin adhesive gel, a foundation, a lipstick, a mascara and a makeup base. More specifically, Etc., and may include various conventional auxiliary agents and carriers suitable for each of these formulations and well known in the art.

In another aspect of the present invention, there is provided a method for producing extracts of Aspergillus oryzae, Leaves of Aspergillus oryzae, and extracts of Edelweiss complex, wherein the natural plants are mixed and extracted with hot water, followed by filtration and freeze-drying. More specifically, it is mixed at a weight ratio of 1: 15-15: 1 to 15: 1, preferably 1: 10: 1 to 10: 1 to 10: , And more preferably from the range of from 5: 1 to 5: 1 to 5: 1: 5: 1: 5; Heating the mixed natural plant at 80 to 120 캜 to obtain an extract; Filtering the extract with a filter, preferably with a 0.4 to 1.5 μm filter to obtain a filtrate; And a step of concentrating and freeze-drying the filtrate.

Hereinafter, the present invention will be described in more detail with reference to examples. It is to be understood that the following examples are for the purpose of illustration only and are not intended to limit the scope of the present invention, and ordinary variations of the present invention are possible within the scope of the present invention.

Example

Example  One

After adding 16.6 g each of 1: 1: 1 to the extraction chamber, 950 g of water is added to the extraction portion, and extraction is carried out at 80 ° C for 2 hours. The thus-extracted extract was filtered with a 0.8 μm and 0.45 μm filter to obtain a filtrate. The filtrate was concentrated at 60 ° C. or lower using a rotary evaporator (NE-series, Eyela, Japan) The concentrate was freeze-dried at -21 ° C and freeze-dried at -80 ° C for 48 hours using a freeze-dryer (Bondiro, Ilshin, Korea) to obtain extracts of Bombyx mori. The thus obtained product was used in the following test examples.

Comparative Example  One

After adding 50 g of sample to the extraction chamber, 950 g of water is added to the extraction portion and extraction is carried out at 80 ° C for 3 hours. The thus-extracted extract was filtered with a 0.8 μm and 0.45 μm filter to obtain a filtrate. The filtrate was concentrated at 60 ° C. or lower using a rotary evaporator (NE-series, Eyela, Japan) The concentrate was freeze-dried at -21 ° C and freeze-dried at -80 ° C for 48 hours using a freeze dryer (Bondiro, Ilshin, Korea) The thus obtained product was used in the following test examples.

Comparative Example  2

After adding 50 g of samarium bamboo leaf sample to the extraction chamber, 950 g of water is added to the extraction portion and extraction is carried out at 80 ° C. for 3 hours. The thus-extracted extract was filtered with a 0.8 μm and 0.45 μm filter to obtain a filtrate. The filtrate was concentrated at 60 ° C. or lower using a rotary evaporator (NE-series, Eyela, Japan) The concentrate was freeze-dried at -21 ° C and freeze-dried at -80 ° C for 48 hours using a freeze dryer (Bondiro, Ilshin, Korea) The thus obtained product was used in the following test examples.

Comparative Example  3

After adding 50 g of marigold sample to the extraction chamber, 950 g of water is added to the extraction portion and extraction is carried out at 80 ° C for 3 hours. The thus-extracted extract was filtered with a 0.8 μm and 0.45 μm filter to obtain a filtrate. The filtrate was concentrated at 60 ° C. or lower using a rotary evaporator (NE-series, Eyela, Japan) The concentrate was freeze-dried at -21 ° C and freeze-dried at -80 ° C for 48 hours using a freeze dryer (Bondiro, Ilshin, Korea) The thus obtained product was used in the following test examples.

Test Example

Test Example  1 - Antioxidant effect measurement test

The free radical scavenging test and the active oxygen scavenging test were carried out in order to confirm the antioxidant effect of the extract of the present invention,

The free radical scavenging test uses the fact that the absorbance of stable DPPH exhibits the maximum absorbance at 540 nm. As the free radical DPPH is erased by the sample and becomes transparent color in purple, that is, as the free radical scavenging rate is increased, And the absorbance of the sample was decreased.

First, 1 mL of the above-mentioned Examples and Comparative Examples were diluted to a suitable concentration in methanol and mixed at 1 mL of 0.1 mM 2,2-diphenyl-1-picryl-hydrazyl radical (DPPH, Sigma) After incubation, the absorbance was measured at 540 nm using a microplate reader (UVT-06685, Thermo max, USA).

In the free radical scavenging test, 1 ml of DPPH and 1 ml of methanol were added to the control group, and 1 ml of methanol and 1 ml of the sample were added to obtain the respective color correction values for the sample and the control group.

The free radical scavenging ratios were calculated using the following equation (1). In Table 1, FSC ₅₀ is the concentration of the sample required to eliminate 50% of the free radicals, and the smaller the value, the higher the antioxidant activity.

In the active oxygen scavenging test, the change of absorbance by the oxidation of nitroblue tetrazolium (NBT) by reactive oxygen species using active oxygen generation by enzyme reaction of xanthine / xanthine oxidase (Sigma) By measuring, it is possible to know the scavenging ability of the active oxygen.

Add 0.1 mL of Na ₂ CO ₃ , 0.1 mL of xanthine (Sigma), 0.1 mL of ethylenediamine tetraacetic acid (EDTA), 0.1 mL of bovine serum albumin (BSA, Sigma), 0.1 mL of NBT and 0.1 mL of the sample and add to the vortex mixer (Type 37600 Mixer, After incubation at 25 ° C for 10 minutes, 0.1 ml of xanthine oxidase was added and reacted at 25 ° C for 20 minutes. The reaction was stopped by adding 6 mM CuCl ₂ to the microplate reader (UVT-06685, Thermo Absorbance was measured at a wavelength of 540 nm by using a spectrophotometer.

The control group in the active oxygen scavenging activity test was prepared by adding the third distilled water instead of the sample solution and measuring by the same method and adding the third distilled water instead of the xanthine oxidase solution to obtain the respective color correction values for the extracted sample and the control group.

The active oxygen scavenging ratio is numerically calculated using Equation (2), and is shown in Table 1 below. In Table 1, the IC ₅₀ is a sample concentration required to eliminate 50% of active oxygen, and a smaller value means that the antioxidant activity is stronger.

The samples used in the test were the extracts of Bombyx mori, Bacillus thuringiensis and Bombyx mori extracts obtained in Example 1 and the extracts of Bombyx mori, Sambusae leaf mackerel or Momordicae japonica extract obtained in Comparative Examples 1 to 3, and in order to compare the antioxidant effect, And the results are shown in Table 1.

As shown in Table 1, the antioxidative effect of the extract was highest in the extracts obtained from extracts of Mulberry, Sangjiwoobyum or Mamallifolia, and the antioxidative activity of the extracts of Mulberry, Mulberry leafhopper and Mulberry extract were superior. In addition, the free radical scavenging activity was similar to that of BHT, a strong synthetic antioxidant used as a positive control.

Test Example  2 - Moisturizing effect  Measurement test

The measurement of the moisturizing effect was carried out by using a capacitance measurement method using a water level measuring device (WSK-P500U, Inforward., Inc, Japan), and by using the 1% aqueous solution samples of Examples and Comparative Examples and various studies, hyaluronic acid acid sodium salt, Sigma). Immediately prior to the start of the experiment, the lateral oysters were allowed to adjust for 20 minutes in a constant temperature and humidity chamber (22 ° C, 40 to 60% relative humidity) to maintain the moisture content of the skin. Then, yusubun meter (WSK-P500U, Inforward. Inc, Japan), the patient in order to minimize the error due After measuring the initial water retention of each sample application site, the samples 2.0 ㎎ / ㎝ ² to forearm oyster side using With different positions of the sample to be applied. The skin moisture content was first measured at the time of applying 10 minutes, 30 minutes, 60 minutes, 90 minutes and 120 minutes, and the skin moisture was again measured after 2 hours. The position of each measurement was repeated three times and the skin moisture content was calculated by the average value, and it is shown in Table 2.

As a result, it was confirmed that all the samples exhibited a higher moisturizing effect than the purified water. Among them, the 1% aqueous solution of the extracts of Bombyx mori, Bacillus thuringiensis extract and Mallipyk complex extract obtained in Example 1 was found to be a 1% aqueous solution of hyaluronic acid And a similar effect was observed, and it was confirmed that the moisturizing effect was the best in the samples.

Test Example  3 - After irradiation with ultraviolet rays, MMP -1 expression inhibition evaluation test

Enzyme immunoassay (ELISA) was performed as described below to assay the inhibition of MMP-1 expression by measuring the concentration of MMP-1 after UV irradiation and sample addition of the extracts of the present invention.

UVA is irradiated to human dermal fibroblasts at an energy of 5 J / cm < 2 > using a UV chamber. Ultraviolet irradiation dose and incubation time were established by preliminary experiment to maximize MMP expression level in fibroblasts. Negative controls were wrapped in silver foil and kept in the UVA environment for the same amount of time. UVA emission was measured using a UV radiometer. The UVA-irradiated cells are irradiated with UVA, and then cultured for 24 hours in the medium containing the examples and the comparative examples. The medium is recovered and coated on the 96-well plate. The primary antibody (MMP-1 (Ab-5) monoclonal antibody and MMP-2 (Ab-3) monoclonal antibody) is treated and reacted at 37 ° C for 60 minutes. After incubation for 60 minutes with a secondary antibody, anti-mouse IgG (alkaline phosphatase conjugated), alkaline phosphatase substrate solution (1 mg / ml ρ-nitrophenyl phosphate in diethanolamine buffer) was reacted at room temperature for 30 minutes. Absorbance is measured at 405 nm using a reader (UVT-06685, Thermo max, USA). In order to evaluate inhibition of MMP-1 expression, retinol, which is known to have excellent inhibitory effect on MMP-1 expression, was set as a comparative group, and the results are shown in Table 3 below.

As shown in the above Table 3, the extracts of Mulberry, Mulberry leafhopper and Mulberry extract showed superior inhibitory effects on MMP-1 expression, which is superior to the control of retinol. In addition, the inhibition rate of retinol was similar or higher than that of retinol.

Test Example  4 - Cell Protection against UV Irradiation  Increase effect test

In order to examine the effect of the present invention on the protection of cells against UV irradiation, the following experiment was conducted as follows.

Fibroblasts were placed in 24-well test plates at 1 × 10 ⁵ cells for 24 h. Each well was washed once with PBS and 500 uL of PBS was added to each well. After irradiating the fibroblasts with 10 mJ / cm ² of ultraviolet light using an ultraviolet B (UVB) lamp (Model: F15T8, UVB 15 W, Sankyo Dennki, Japan), PBS was removed and cultured in a cell culture medium Was added. After treating the extracts of Bombyx mori, Bacillus thuringiensis and Bombyx mori extracts (Example 1), Bombyx mori extract (Comparative Example 1), Trilobarbital extract (Comparative Example 2) or Marigold extract (Comparative Example 3) Lt; / RTI > After 24 hours, the medium was removed, and 500 μL of the cell culture medium and 60 μL of the MTT solution (2.5 mg / mL) were added to each well, followed by culturing in a 37 ° C. CO ₂ incubator for 2 hours. The medium was removed and 500 μL of isopropanol-HCl (0.04 N) was added. Cells were lysed by shaking for 5 minutes and 100 μL of the supernatant was transferred to a 96-well test plate and absorbance at 565 nm was measured using a microplate reader (UVT-06685, Thermo max, USA). The cell survival rate (%) was measured by the formula (3), and the cell protection increase rate against the ultraviolet ray was calculated by the formula (4) and is shown in the following table.

As shown in the above Table 4, it was found that the cell protection against UV rays was effective when the extracts of Bombyx mori, Bacillus thuringiensis and Bombyx mori extracts were more complex than the extracts of Bombyx mori.

Test Example  5 - keratinocyte cell line From HaCaT Retinoic acid  or Sodium lauryl sulfate Sodium Lauryl Sulphate , SLS ) Stimulation effect test by suppression of prostaglandin E2 and α-MSH (Melanocyte stimulating hormone) secretion by stimulation

1% fetal bovine serum (FBS, available from Gibco, USA) and 1% fetal calf serum (cell line name: HaCaT available from Dr. NE Fusenig, Deutsches Krebsforschungszentrum, Heidelberg, Germany) (96-well plate) was cultured in DMEM (Dulbecco's Modified Eagle Medium, available from Lonza, USA) containing penicillin-streptomycin (Gibco, USA) ⁴ cells / well and then cultured at 37 ° C under 5% CO ₂ for 24 hours. After the culture medium was removed, the cells were washed once with 200 μl of phosphate buffered saline (PBS), and 200 μl of a DMEM medium containing no fetal bovine serum was added thereto and cultured for 24 hours. Then, the following Examples and Comparative Examples were treated with 0.01% concentration of DMEM containing 1% fetal bovine serum for 10 minutes, and then 100 μM of retinoic acid (Retinoic acid, available from Sigma-Aldrich, USA) or 10 ppm of sodium (Hereinafter referred to as " substance-treated group ") with lauryl sulfate (sodium lauryl sulfate, available from Sigma-Aldrich, USA) for 24 hours. Prostaglandin E2 (PGE ₂₎ ELISA kit (available processing: R & D system, USA) by taking the culture medium was measured prostaglandin E2 50㎕ glass inhibitory effect of retinoic acid or sodium lauryl sulfate using a, the inhibitory ability was standard A standard curve was drawn based on the absorbance of rh PGE _{2 to determine} the equation between the absorbance and the reference material, and the absorbance of the treated material was substituted to obtain the secretion amount of PGE ₂ . Then, the stimulation relaxation rate was evaluated by the following equation (5), and the results are shown in Table 5 below.

The keratinocyte was cultured in the same manner as described above and cultured in the same manner as in Example and Comparative Example. 50 μl of the culture was taken and α-MSH (Melanocyte stimulating hormone) was used instead of the prostaglandin E2 (PGE ₂ ) ELISA kit. The inhibitory effect of retinoic acid or sodium lauryl sulfate on the α-MSH activity was measured using an EIA-ELISA kit (available from Phoenix Pharmaceutical Inc. USA). The inhibitory activity was measured using standard The curve was drawn to obtain the reaction equation between the absorbance and the standard substance, and the absorbance of the treated substance was substituted to obtain the secretion amount of? -MSH. The stimulation relaxation rate was evaluated by the following equation (6), and the results are shown in Table 5 below.

The results of the above Table 5 show that the inhibitory effect of prostaglandin E2 on secretion was significantly reduced in the case of treating the extracts of Bombyx mori, Bacillus thuringiensis and Bombyx mori copra prepared in Example 1 compared with the case of treating the single extract prepared in Comparative Examples 1 to 3 Inhibiting the secretion of prostaglandin E2 to a remarkably high level.

In addition, the secretion performance of? -MSH was significantly higher in the case of treating the extract of Bombyx mori, Bacillus thuringiensis and Bombyx mori copra prepared in Example 1 compared to the case of treating the single extract prepared in Comparative Examples 1 to 3 And the inhibition of the secretion of [alpha] -MSH at the < RTI ID = 0.0 >

Test Example  6 - According to the invention Cosmetics  Skin Soothing Test of Composition

In order to examine the skin irritation mitigation effect (skin soothing effect) when applying the cosmetic composition according to the present invention, skin irritation mitigation test was conducted on 20 men and women in their 20s and 40s.

L-lactic acid was added to the four kinds of extracts prepared in Example 1 and Comparative Examples 1 to 3 for 10 days as a stimulus source, and a relaxation effect on stimulation caused by lactic acid Respectively. Every morning and evening, they were asked to use the questionnaire to indicate whether they were stimulated or not. The results of the test were evaluated once per day and divided into subjective stimuli such as tingling or itching and objective stimuli such as skin erythema and edema. The results are shown in Table 6 below.

As can be seen from Table 6, it was found that the extracts of Bombyx mori, Bacillus thuringiensis and Bombyx mori extract prepared in Example 1 showed the best mitigation effect of skin irritation induced by 5% lactic acid.

Examples of formulations of the present invention are a soft lotion, a convergent lotion, a nutritional lotion, a massage cream, an essence and a pack, but the formulation of the cosmetic composition of the present invention should not be construed as being limited thereto. Lt; / RTI > is possible.

[Formulation Example 1] Flexible longevity

[Formulation Example 2] Convergent lotion

[Formulation Example 3] Nutritional lotion

[Formulation Example 4] Essence

[Formulation Example 5] Pack

Claims (11)

The present invention relates to a cosmetic composition comprising as an active ingredient an extract of Aspergillus oryzae. The cosmetic composition as set forth in claim 1, wherein the granulosa extract, Sanko leafworm, and Japanese mackerel are mixed in an amount of 1 to 15: 1 to 15: 1 to 15 by weight, and the combined extract is used as an active ingredient. The cosmetic composition according to claim 1, wherein the cosmetic composition has an antioxidative activity effect. The cosmetic composition according to claim 1, wherein the cosmetic composition has a cytoprotective effect. The cosmetic composition according to claim 1, wherein the cosmetic composition has skin moisturizing effect. The cosmetic composition according to claim 1, wherein the cosmetic composition is skin-soothing. [Claim 2] The cosmetic composition according to claim 1, wherein the extracts of Ganpungcho, Samjungwoo lobster, and Mamallipu complex are hydrothermal extracts or solvent extracts. The method of claim 7, wherein the solvent is selected from the group consisting of water, fermented alcohol, ethanol, methanol, butanol, propanol, ethyl acetate, isopentyldiol, glycerin, ethylene glycol, propylene glycol, butylene glycol, chloroform, dichloromethane, And at least one solvent selected from the group consisting of acetonitrile, petroleum ether and diethyl ether. [Claim 2] The cosmetic composition according to claim 1, wherein the granular extracts of Ganoderma lucidum, Lycopersicon esculentum, and Pseudomonas sp. Are contained in an amount of 0.001 to 10 wt% based on the total weight of the cosmetic composition. The cosmetic composition according to claim 1, wherein the antioxidant, cell protection, skin moisturizing and skin clearing cosmetic composition is at least one selected from the group consisting of lotion, cream, essence, cleansing foam, cleansing water, pack, patch, body wash, body lotion, body oil, Wherein the cosmetic composition is prepared by any one of the following formulations: a rinse, a hair conditioner, a hair gel, a skin adhesive gel, a foundation, a lipstick, a mascara and a makeup base product. A step of mixing phytophthora balsa herbaceousum, three leaves groundworm, and mustard seeds;
Heating the mixture at 80 to 120 캜 to obtain an extracted extract;
Filtering the extract with a filter to obtain a filtrate; And
And concentrating and lyophilizing the filtrate to obtain a combined extract. ≪ RTI ID = 0.0 > 8. < / RTI >