KR101950384B1 - Cosmetic composition comprising extracts of Trachelospermum jasminoides, Coptis chinensis and Sophora Japonica as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition containing, as an active ingredient, at least two plant extracts selected from the group consisting of a pineal gland extract, a yellowtail extract and a pictorial tree extract. The plant extracts of at least two selected from the group consisting of a hairy line extract, a Rhodiola extract and a conifer tree extract according to the present invention have an anti-aging effect on the skin. Specifically, the extract has the effect of promoting the aquaporin biosynthesis, the inhibitory effect of MMP-1 expression, Promoting effect, inhibiting melanin formation, inhibiting miR-378b expression, promoting miR-181a expression, promoting miR-340 expression, promoting dermal keratinocyte differentiation, enhancing collagen biosynthesis, improving skin barrier function, , Whitening effect, skin wrinkle improving effect, skin elasticity improving effect, and dermal density improving effect, and thus various functional cosmetic materials can be provided.

Description

[0001] The present invention relates to a cosmetic composition containing, as an active ingredient, an extract of Alaska pollack, an extract of Alaska pollack, a leaf extract of Alaska pollack,

The present invention relates to a cosmetic composition comprising a pine needle extract, a Rhodiola extract and a conifer tree extract as an active ingredient.

A variety of physical and chemical changes occur in human skin during the aging process. It is divided into intrinsic aging and photo-aging depending on the cause, and researches on it have been actively conducted. Free radicals may be activated due to ultraviolet rays, stress, disease state, environmental factors, wounds, age and the like. When such state is deepened, the antioxidant defense network existing in the living body is destroyed, Thereby promoting aging.

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

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.

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

Korean Patent Laid-Open Publication No. 2001-0018657 relates to a cosmetic composition containing an extract of Lactobacillus japonica. Korean Patent Laid-Open Publication No. 2016-0084900 relates to a cosmetic composition for skin whitening containing an inorganic ring composed of extracts of Wang Lil, Satsu Oil, and Baekjak extract.

The present inventors have studied the applicability of cosmetics to various natural products that have not been known until now. As a result, the present inventors have found that the extracts of the present invention can be used to produce MMP-1 production inhibitory effect, collagen biosynthesis effect, Enhancing effect of aquaporin biosynthesis, promoting biosynthesis of hyaluronic acid, promoting dermal keratinocyte differentiation, inhibiting melanin production, moisturizing effect, whitening effect, skin barrier function improving effect, skin wrinkle improving effect, skin elasticity improvement effect, And anti-aging effect on skin, it was found that the efficacy as a cosmetic product can be expected because the effect is very excellent.

It is an object of the present invention to provide a method of inhibiting MMP-1 production, collagen biosynthesis effect, hyaluronic acid biosynthesis promotion effect, aquaporin biosynthesis promotion effect, skin keratinocyte differentiation promotion effect, melanin production The present invention is to provide a cosmetic composition which exhibits an inhibitory effect, a moisturizing effect, a whitening effect, a skin barrier function improving effect, a skin wrinkle improving effect, a skin elasticity improving effect, a dermis tightness improving effect and a skin antiaging effect.

It is another object of the present invention to provide a cosmetic composition exhibiting superior efficacy and efficacy through preparation of supercritical, ultrasonic and fermented extracts and purification to maximize the efficacy of the pine needle extract, Huaxia extract, and coniferous tree extract.

It is another object of the present invention to provide a makeup method using the cosmetic composition containing the pine needle extract, Huanglian extract, and coniferous tree extract.

The present invention relates to a cosmetic composition containing two or more plant extracts selected from the group consisting of a pineal gland extract, a yellowtail extract, and a pictorial tree extract. And more preferably to a cosmetic composition containing a wooly husk extract, a thyme extract and a conifer tree extract.

Preferably, the cosmetic composition has an effect of improving the skin barrier function.

Preferably, the cosmetic composition has skin moisturizing effect.

Preferably, the cosmetic composition has an effect of improving skin elasticity

Preferably, the cosmetic composition has an effect of improving skin wrinkles.

Preferably, the cosmetic composition has a skin whitening effect.

Preferably, the plant extract is contained in an amount of 0.001 to 30.0% by weight based on the total weight of the cosmetic composition.

Preferably, the plant extract is selected from the group consisting of (a) water, an anhydrous or lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, dichloromethane, Hexane and mixtures thereof, or (b) by extraction with fermentation or natural fermentation from a microorganism.

Preferably, the microorganism is Lactobacillus plantarum ATCC 10012.

The plant-mixed extracts selected from the group consisting of fur extract, Huaxia extract and Conifer tree extract according to the present invention were found to be effective in promoting aquaporin biosynthesis (Experimental Example 1), skin keratinocyte differentiation promoting effect (Experimental Example 2), melanin production inhibition (Example 5), collagen biosynthesis effect (Experimental Example 6), miR-378b expression inhibitory effect (Experimental Example 7), collagen synthesis inhibitory effect (Experimental Example 3), and hyaluronic acid expression promoting effect (Experimental Example 4) (Experimental Example 9), skin barrier function improving effect (Experimental Example 10), moisturizing effect (Experimental Example 11), whitening effect (Experimental Example 12), miR-181a expression enhancing effect (Experimental Example 8) , Skin elasticity improving effect (Experimental Example 13), skin wrinkle improving effect (Experimental Example 14), and dermal density improving effect (Experimental Example 15).

1 is a graph showing the effect of promoting biosynthesis of hyaluronic acid in a plant extract according to the present invention.

Fossil fuels used as active ingredients in the cosmetic composition of the present invention are represented by the scientific name Trachelospermum jasminoides , and are also called tinnitals such as fur, pine, rhizome, and fir. The fur line is a vine plant in the middle part of the Kangwon tree, and the length is 2 ~ 3m. Leaves are opposite and long oval. Leaf length is 4 ~ 8cm, width is 2 ~ 5cm and tip is sharp. From May to June, a yellowish white flower blooms at the tip of the branch. The fruit is 10 ~ 15cm in length and has a spindle shape. They are distributed in southern Korea, Japan, and China. In the present invention, the wooly crest extract is obtained by extracting from various organs or parts (e.g., leaves, branches, flowers, roots, stems, etc.) of wooly threads, preferably an extract obtained from the outposts or stems, Preferably an extract obtained from the outpost.

Coptis Chinensis is used as an active ingredient in the cosmetic composition of the present invention, and its phylum is also referred to as Coptis Chinensis . Chrysanthemum is an evergreen perennial plant of the family Ranunculaceae, which grows in Korea, Japan, China and other places. It grows in ecological land of shrubs in the mountains. The underground stem is thick and spreads to the side. It has many beard roots, 4 ~ 5 roots at the end of the stem, and the length is 10 ~ 27cm. The leaf is a compound leaf with three small leaves. The small leaf is a little hard, sharp and sharp. The petiole of the petiole is a sheath with sawteeth on the edge of the leaf. In April ~ April, two to three white flowers bloom on the flower stalk which grew to about 10cm in diameter. Flowers have blooming flowers and male flowers. The white calyx is five to six long and looks like a petal. Petals are 8-15 small, linear. There are many stamens and 9 ~ 16 pistils. An ovary sack grows, and a fenugreek (袋 果) is lined on it. It is named after the stem and the stem of the underground stem are dark yellow. The Rhizome extract means an extract obtained from various organs or parts (eg, leaves, branches, flowers, roots, stems, etc.) of Rhizoctonia, preferably an extract obtained from a plant or root, .

The pictured tree used as an active ingredient in the cosmetic composition of the present invention is Sophora japonica . Painted wood is rosemary bean and sorghum, and the height of the tree grows up to about 25m and the branches spread widely. Small branches are green and smell when cut. Leaves are alternate and once folded leaves. Small leaves are 7 ~ 17 eggs, ovate or egg-shaped, oval on the back, with small petiole on the back, there are loose hairs. The flowers bloom in August and run on cones. The shape of the fruit that opens after the flower is formed is pod-like, and the shape of the round seeds is unique. It is about 5-8cm in length, and the seeds of the seeds are narrowed down and sagged downward. The bud is called 槐花 or 미米, and the fruit is called 槐 实, and it is used as medicinal medicine in Oriental medicine. The fruit is a cone with a cylindrical or bead shape. It is used for arteriosclerosis and hypertension, and for making beer and paper yellow. The sprouts are used for hemorrhoid treatment, with branches and bark. It grows in the mountains of Korea, and uses garden or wood as furniture. In the present invention, the Painted Tree Extract is obtained by extracting from various organs or parts (e.g., leaves, branches, flowers, roots, stems, fruits, etc.) of a painting tree, preferably an extract obtained from a plant or stem , And most preferably from an outpost.

The present invention relates to a cosmetic composition comprising a plant extract containing two or more, preferably all three, selected from the group consisting of a pineal gland extract, a thyme extract and a pictorial tree extract.

In case of mixing two kinds of the extracts of Wooly Rhizome, Rhodiola extract and Concha tree, the plant extract to be mixed may be mixed at a weight ratio of 1: 5 to 5: 1. 1 to 5: 1 to 5: 1, preferably 1: 3: 1 to 3: 1 to 3, in the case of mixing the three kinds of modes of the pine needle root extract, Preferably in a weight ratio of 1: 1: 1. In the case of using the pine root extract, Rhodiola extract, and coniferous tree extract outside the above range, synergistic effects obtained by mixing the components as in the case of using each alone can not be obtained.

The plant extracts according to the present invention can be prepared by using conventional solvents known in the art, that is, under the conditions of ordinary temperature and pressure, using conventional solvents (A) water, anhydrous or lower alcohol having 1 to 4 carbon atoms (for example, methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, di Extraction can be carried out using a solvent extraction method using a solvent selected from the group consisting of ethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane, and mixtures thereof.

In the present invention, a solvent extraction method using an extraction solvent is preferably used. Preferably ethanol, 70% (v / v) ethanol or water, and most preferably 70% (v / v) ethanol.

It will be apparent to those skilled in the art that the extracts of Trichinella crassa, Rhodochrosite extract and Coniferous tree extract of the present invention can be obtained not only by the above-mentioned extraction solvents but also by other extraction solvents, which exhibit substantially the same effects.

In addition, the plant extracts according to the present invention (hairy roots extract, Huangliao extract, and conifer tree extract) also include fermented extracts. The fermented extracts of Curcuma longa, Rhododendron japonica extract and Coniferous tree extract can be prepared as follows. Each plant extract is finely crushed to a size of about 100 to 500 mesh, and 1 to 30% by weight of a plant extract is added to a conventional microorganism culture solution, and lactic acid bacteria are added in an amount of 10,000 to 10,000,000 cfu / mL. The culture temperature is 30 to 37 ° C. The pH is 5 to 7 and is usually anaerobically cultured for about 24 to 72 hours. Which can be obtained through aging and filtration. The lactic acid bacteria used in the present invention may preferably be Lactobacillus plantarum ATCC 10012.

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

Therefore, the cosmetic composition of the present invention comprising two or three kinds of wooly crestal extract, Rhodiola extract, and pictorial tree extract is for preventing skin aging. Specifically, prevention of skin aging can be effectively prevented by inhibiting the production of MMP-1, inhibiting miR-371b expression, promoting miR-181a expression, inhibiting miR-340 expression, promoting hyaluronic acid expression, and promoting aquaporin biosynthesis A skin barrier function, a skin wrinkle improving effect, a skin elasticity improving effect, and a dermatome tightness improving effect, which are effective in improving the dermal keratinocyte differentiation promotion effect, melanin generation inhibitory effect, moisturizing effect, whitening effect, skin barrier function.

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 .

Production Example 1: Production of a wooly husk extract

100 g of shredded shredded pods were filtered with 3 times of reflux with 70% (v / v) aqueous ethanol solution for 5 hours, followed by cooling, followed by filtration through Whatman # 4 filter paper. The filtered extract was concentrated under reduced pressure and freeze-dried at 50 DEG C or less to obtain 17.4 g of a pine needle extract.

Production Example 2: Preparation of Rhodiola extract

After shredding, 100 g of shrunken roots were refluxed with 70% (v / v) aqueous ethanol solution for 3 hours each time, cooled down, and filtered through Whatman # 4 filter paper. The filtered extract was concentrated under reduced pressure and freeze-dried at 50 DEG C or lower to obtain 19.7 g of Rhodiola extract.

Production Example 3: Production of Painted Tree Extract

100g of the shrubs and shrubs were washed with a 70% (v / v) aqueous ethanol solution for 3 hours, refluxed, and filtered through Whatman # 4 filter paper. The filtered extract was concentrated under reduced pressure and freeze-dried at 50 ° C or lower to obtain 18.1 g of a tree extract.

Production Example 4: Preparation of Fermented Extract of Fur Seed Extract

Production Example 4 was prepared by fermenting lactic acid bacteria in Production Example 1 as a fermentation extract of a wooly husk extract. Lactobacillus MRS broth (DIFCO, USA) was used as a culture medium for lactic acid bacteria, and 1 to 30% by weight, preferably 10% by weight, of the wooly crest extract of Preparation Example 1 was added thereto.

The Lactobacillus MRS medium was inoculated with 1,000,000 cfu / ml of Lactobacillus plantarum ATCC 10012 and cultured at 37 ° C for 72 hours. After the incubation, the culture was centrifuged to remove the bacteria, and the fermented product of the furrow extract was added thereto. Ethanol was added to the final 70% (V / V), refluxed three times for 5 hours, And filtered through Whatman # 4 filter paper. The filtered fermented extract was filtered with 0.25 μm filter paper, and then concentrated under reduced pressure and freeze-dried at 60 ° C or lower. Ethanol and butyleneglycol were dissolved in at least one solvent so as to contain 0.001 to 70% by weight of the thus obtained reduced-pressure concentrate or lyophilized product to obtain a fermented extract of Fur Seed Extract (Preparation Example 4).

Preparation Example 5: Preparation of fermented extract of Rhodiola extract

Production Example 5 was prepared by fermenting lactic acid bacteria of Preparation Example 2 as a fermented extract of Huangyan extract. Lactobacillus MRS broth (DIFCO, USA) was used as a culture medium for lactic acid bacteria, and 1 to 30 wt.%, Preferably 10 wt.

The Lactobacillus MRS medium was inoculated with 1,000,000 cfu / ml of Lactobacillus plantarum ATCC 10012 and cultured at 37 ° C for 72 hours. After culturing, the culture broth was centrifuged to remove the bacteria to obtain a fermented product of Rhodiola extract. Ethanol was added to the final 70% (v / v), refluxed for 3 hours for 5 hours, And filtered through Whatman # 4 filter paper. The filtered fermented extract was filtered with 0.25 μm filter paper, and then concentrated under reduced pressure and freeze-dried at 60 ° C or lower. The fermented extract of Rhodiola extract was obtained by dissolving the thus obtained reduced pressure concentrate or lyophilizate in at least one solvent in purified water, ethanol and butylene glycol so as to contain 0.001 to 70% by weight (Preparation Example 5).

Production Example 6: Production of a woody fermented extract

Production Example 6 was prepared by fermenting lactic acid bacteria in Production Example 1 as a fermented extract of Coniferous tree extract. Lactobacillus MRS broth (DIFCO, USA) was used as a culture medium for lactic acid bacteria, and 1 to 30% by weight, preferably 10% by weight, of the extract of Painting Tree of Preparation Example 3 was added thereto.

The Lactobacillus MRS medium was inoculated with 1,000,000 cfu / ml of Lactobacillus plantarum ATCC 10012 and cultured at 37 ° C for 72 hours. After the culture, the culture was centrifuged to remove the bacteria, and the resulting extract was subjected to refluxing for 3 hours for 5 hours, followed by addition of ethanol to the final 70% (V / V) And filtered through Whatman # 4 filter paper. The filtered fermented extract was filtered with 0.25 μm filter paper, and then concentrated under reduced pressure and freeze-dried at 60 ° C or lower. Ethanol and butyleneglycol were dissolved in at least one solvent to obtain 0.001 to 70% by weight of the thus obtained reduced-pressure concentrate or lyophilized product.

Manufacturing example  Preparation of 7 to 18: Preparation of two mixed plant extracts

The plant extracts and fermentation extracts of Preparation Examples 1 to 6 thus prepared were mixed so that the weight ratio of the two species was 5: 1 to 1: 5. Production Example 7 is a 1: 3 mixture of Production Example 1 and Production Example 2, Production Example 8 is a 3: 1 mixture of Production Example 1 and Production Example 2, Production Example 10 is a 1: 3 mixture of Production Example 1 and Production Example 3, Production Example 11 is a 3: 1 mixture of Production Example 1 and Production Example 3, Production Example 12 is Production Example 1 and Production Example 3, Production Example 13 is a 1: 3 mixture of Production Example 2 and Production Example 3, Production Example 14 is a 3: 1 mixture of Production Example 2 and Production Example 3, Production Example 15 is a 1: 2, and Production Example 3, Production Example 16 is a 1: 1 mixture of Production Example 4 and Production Example 5, Production Example 17 is a 1: 1 mixture of Production Example 4 and Production Example 6, 18 is a 1: 1 mixture of Preparation Example 5 and Preparation Example 6.

Manufacturing example  Preparation of 19 to 32: Preparation of three kinds of mixed plant extracts

The plant extracts and the fermentation extracts of the above-mentioned Production Examples 1 to 6 were mixed so that the weight ratio of the three species was 1: 5: 1 to 5: 1 to 5. Production Example 19 is a 5: 1: 1 mixture of Production Example 1, Production Example 2 and Production Example 3, Production Example 20 is a 3: 1: 1 mixture of Production Example 1, Production Example 2 and Production Example 3, 21 is a 1: 1: 1 mixture of Production Example 1, Production Example 2 and Production Example 3, Production Example 22 is a 1: 1: 3 mixture of Production Example 1, Production Example 2 and Production Example 3, 1: 5 mixture of Production Example 1, Production Example 2 and Production Example 3, Production Example 24 is a 1: 3: 1 mixture of Production Example 1, Production Example 2 and Production Example 3, Production Example 26 is the 5: 1: 1 mixture of Production Example 4, Production Example 5 and Production Example 6, Production Example 27 is the mixture of Production Example 4 and Production Example 5, Production Example 28 is a 1: 1: 1 mixture of Production Example 4, Production Example 5 and Production Example 6, Production Example 29 is a mixture of Production Example 4 and Production Example 6, 5 and Preparation Example 6, Production Example 30 is a 1: 1: 5 mixture of Production Example 4, Production Example 5 and Production Example 6 , Production Example 31 is a 1: 3: 1 mixture of Production Example 4, Production Example 5 and Production Example 6, and Production Example 32 is a 1: 5: 1 mixture of Production Example 4, Production Example 5 and Production Example 6. [

Experimental Example  One: Furring  extract, goldthread  Extract and Painted Tree Extract Aquaporin  3 Biosynthesis Promotion Experiment

In order to measure the acceleration of the formation of aquaporin 3 in the samples prepared in Preparation Examples 1 to 32, the following experiment was conducted. Human keratinocytes were inoculated on a 24-well plate (Falcon, USA) at a concentration of 1 × 10 ⁴ cells / well, and then cultured for one day to stabilize the cells. And cultured in serum-free medium for 2 days. After incubation, the supernatant was taken and the absorbance at 460 nm was compared using aquaporin 3 assay kit (Aquaporin 3 ELISA KIT, Gill Blood Group, USA) to compare the degree of production of aquaporin 3 to determine the effect of each sample on the aquaporin biosynthesis Respectively.

As can be seen from the results of Table 1, the extracts of Production Examples 1 to 32 of the present invention promote the biosynthesis of aquaporin 3 in dermal keratinocytes. In particular, the effects of Preparation Examples 7 to 18, which are two kinds of preparations, are superior to those of Production Examples 1 to 6, which are single extracts, and Production Examples 19 to 32, which are three kinds of preparations, have the best promoting effect on biosynthesis of aquaporin 3 .

Experimental Example  2: Fur Seed Extract, goldthread  Promotes the differentiation of skin keratinocytes in extracts and tree extracts

Human keratinocytes were inoculated into 6 well plates. After 24 hours, the cells grown to 70-80% were replaced with serum-free DMEM and cultured for 5 hours. After culturing for 5 hours, Production Examples 1 to 32 were treated with 0.1%. As a comparative example, a solvent (DMSO) was used instead of the production example. After 16 hours of incubation, the cells were washed with PBS, and total RNA was extracted with RNA prep kit (Qiagen, Germany). The amount of RNA was standardized by UV absorbance, and filaggrin, The expression levels of involucrin, keratin 1, and keratin 10 genes were quantified by qRT-PCR. The expression level of the gene was corrected by GAPDH. The results are shown in Table 2.

As shown in Table 2, it can be seen that Production Examples 1 to 32 promote gene expression of differentiation inducing proteins of keratinocytes. The effects of Preparation Examples 7 to 18, which are two kinds of preparations compared to the single extracts of Preparation Examples 1 to 6, are excellent. In particular, Production Examples 19 to 32, which are three kinds of preparations, promote the gene expression of the differentiation inducing protein of keratinocytes It can be confirmed that the effect is remarkably excellent.

Experimental Example  3: Fur Seed Extract, Rhodiola extract  And Painted Tree Extract B16F1  Inhibitory effect of melanin-forming cells on melanogenesis

In Experimental Example 3, in order to examine the whitening effect of the plant extracts of Preparation Examples 1 to 32, the inhibitory effect on melanin formation on B16F1 melanin-forming cells was examined to determine the whitening effect. 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 after the cells were attached, the extracts of Preparations 1 to 32 were treated at a concentration not causing toxicity and cultured for 72 hours. After incubation for 72 hours, the cells were detached with trypsin-EDTA, the number of cells was measured, and the cells were recovered by centrifugation. Quantification of intracellular melanin was carried out with a slight modification of the method of Lotan (Cancer Res., 40: 3345-3350, 1980). Cell pellet was washed once with PBS, and 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt the cells. After centrifugation (3,000 rpm, 10 min), 1N NaOH (10% DMSO) was added to the cell filtrate to dissolve the extracted melanin and the absorbance of melanin was measured at 405 nm with a microplate reader. The inhibition rate (%) of melanin formation was measured. Melanin formation inhibition rate (%) of B16F1 melanin-forming cells was calculated by the following equation (1), and the results are shown in Table 3. [

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

As can be seen from the results of Table 3, it was found that the extracts prepared according to Production Examples 1 to 32 of the present invention inhibited melanin production in B16F1 melanin-forming cells. Production Examples 7 to 18, which are two kinds of preparations as compared with the single extracts of Preparation Examples 1 to 6, inhibit melanogenesis in B16F1 melanin-forming cells, and in particular, Preparation Examples 19 to 32, which are mixtures of three, inhibit melanin production in B16F1 melanin- It is possible to confirm that the generation is greatly inhibited.

Experimental Example  4: Fur Seed Extract, Rhodiola extract  Promoting effect of hyaluronic acid biosynthesis on coniferous tree extract

Human fibroblasts were cultured in DMEM containing 10% FBS at a concentration of 2 × 10 ⁴ cells / well in a 24-well plate. After 24 hours, the cells grown to 70-80% were replaced with serum-free DMEM and cultured for 5 hours. After culturing for 5 hours, Production Examples 1 to 32 were treated with 0.5%. Three days after the culture, the amount of total hyaluronic acid in the medium was measured by using a Hyaluronan Quantikine ELISA kit (R & D biosystem), and the result was shown in FIG.

As can be seen from the results of FIG. 1, it was found that the extracts prepared according to Production Examples 1 to 32 of the present invention promote biosynthesis of hyaluronic acid in fibroblasts. The effects of Preparation Examples 7 to 18, which are two kinds of preparations compared to the single extracts of Preparation Examples 1 to 6, promote the biosynthesis of hyaluronic acid in the fibroblasts, and in particular, Preparation Examples 19 to 32, which are mixtures of three, It can be confirmed that biosynthesis is greatly promoted.

Experimental Example 5: Inhibitory effect on the production of MMP-1 by the extracts of Walnut, Rhodochrosite and Painted Wood

Experimental example 5 measures the effect of inhibiting the production of collagenase, MMP-1, by the extracts obtained in Preparation Examples 1 to 32.

Human normal skin cells were inoculated into a 48-well microplate (Nunc, Denmark) at a concentration of 1 × 10 ⁶ cells per well and cultured in DMEM medium (Sigma, USA) and 37 ° C For 24 hours, and further cultured in serum-free DMEM medium supplemented with the extracts of Preparation Examples 1 to 32 and in serum-free DMEM medium containing no extract for 48 hours as a control group.

After culturing, the supernatant of each well was collected and the amount (ng / ml) of the newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, USA) The inhibition rate (%) was calculated and the results are shown in Table 4.

As shown in the results of Table 5, it was confirmed that the extracts 1 to 32 of the present invention inhibited the production of MMP-1 in a concentration-dependent manner. In particular, Production Examples 7 to 18, which are two kinds of preparations compared to Production Examples 1 to 6, which are single extracts, Production Examples 19 to 32, which are three kinds of mixtures as compared to the two kinds of preparations, produce MMP-1 in fibroblasts . ≪ / RTI >

EXPERIMENTAL EXAMPLE 6. Enhancement of Collagen Synthesis Effect of Walnut Extract, Huang Lien Extract and Art Tree Extract

Human normal epithelial cells, fibroblasts, were inoculated into 48-well microplates at a density of 1 x 10 ⁶ cells per well and cultured in DMEM medium for 24 hours. Serum-free DMEM medium supplemented with the extracts prepared in Preparation Examples 1 to 32 and the control group were further cultured in serum-free DMEM medium without extract for 48 hours. After the incubation, the supernatant of each well was collected, and the amount of procollagen type I C-peptide (PICP) was measured using a collagen kit (Takara, Japan) and converted into ng / Respectively. The increase rate (%) of collagen biosynthesis was calculated according to the following formula (3), and the results are shown in Table 5.

As can be seen from Table 5, it was confirmed that the collagen biosynthesis rate of the extract prepared according to Production Examples 1 to 32 of the present invention increased in a concentration-dependent manner. The collagen biosynthesis of the two kinds of preparations 7 to 18 as compared with the single extracts of Preparation Examples 1 to 6 was promoted, and in particular, it was confirmed that Production Examples 19 to 32, which were three kinds of mixtures, significantly enhanced collagen biosynthesis in fibroblasts .

Experimental Example  7. Wheatgrass extract, Rhodiola extract  And Painted Tree Extract miR -378b expression inhibitory effect

Human normal epithelial cells, fibroblasts, were inoculated into 48-well microplates at a density of 1 x 10 ⁶ cells per well and cultured in DMEM medium for 24 hours. The cells were further cultured in serum-free DMEM medium supplemented with the extract prepared in Preparation Examples 1 to 32 and in a serum-free DMEM medium containing no extract, followed by washing with PBS, and the miRNA prep kit (Qiagen, Germany ). The expression level of miR-378b, an miRNA that inhibits the expression of SIRT6, which regulates the expression of COL1A1, was quantified using qRT-PCR. The expression level of the gene was corrected using SnoRD6. The results are shown in Table 6.

According to Table 6, it was confirmed that the expression of miR-378b in the extracts prepared according to Production Examples 1 to 32 of the present invention decreased in a concentration-dependent manner. The production of miR-378b in fibroblasts was significantly lowered in the two preparations, which were the two kinds of preparations compared to the single extracts of Preparation Examples 1 to 6. In particular, the preparation examples 19 to 32, -378b significantly decreased and the effect of improving the elasticity was excellent.

EXPERIMENTAL EXAMPLE 8. Promoting Expression of miR-181a Expression in Walnut Extract, Hwangryun Extract, and Conifer Tree Extract

Human normal keratinocyte keratinocyte was inoculated into a 48-well microplate at a density of 1 x 10 ⁶ cells per well and cultured in DMEM medium for 24 hours. The cells were further cultured in serum-free DMEM medium supplemented with the extract prepared in Preparation Examples 1 to 32 and in a serum-free DMEM medium containing no extract, followed by washing with PBS, and the miRNA prep kit (Qiagen, Germany ), The amount of miRNA was standardized by UV absorbance, and the amount of expression of miR-181a, a miRNA associated with keratinocyte differentiation, was quantified by qRT-PCR. The expression level of the gene was corrected by SnoRD6. The results are shown in Table 7.

According to Table 7, it was confirmed that the expression of miR-181a in the extracts prepared according to Production Examples 1 to 32 of the present invention increased in a concentration-dependent manner. Production Examples 7 to 18, which are two kinds of preparations as compared with the single extracts of Preparation Examples 1 to 6, increased the expression of miR-181a in keratinocytes, and in particular, Preparation Examples 19 to 32, which are mixtures of three, the expression of miR-181a was greatly increased and it was confirmed that the moisturizing effect was excellent.

Experimental Example  9. Wheatgrass extract, Rhodiola extract  And Painted Tree Extract miR -340 expression promotion effect

B16F1 melanin-forming cells were inoculated on a 48-well microplate at 1 × 10 ⁶ cells per well and cultured in DMEM medium for 24 hours. The cells were further cultured in serum-free DMEM medium supplemented with the extract prepared in Preparation Examples 1 to 24 and in a serum-free DMEM medium containing no extract, followed by washing with PBS, and the miRNA prep kit (Qiagen, Germany ), The amount of miRNA was normalized by UV absorbance, and the expression level of miR-340, an miRNA that inhibits the expression of MITF, was quantified using qRT-PCR. The expression level of the gene was corrected by SnoRD6. The results are shown in Table 8.

As can be seen from Table 8, the expression ratios of miR-340 of the purified extracts prepared in Production Examples 1 to 32 of the present invention were increased in a concentration-dependent manner. The effects of Preparation Examples 7 to 18, which are two kinds of preparations compared to the single extracts of Preparation Examples 1 to 6, were excellent, and in particular, Production Examples 19 to 32, which were three kinds of preparations, significantly increased the expression of miR-340 in melanin- And the whitening effect is excellent.

Formulation Example 1: Preparation of Cosmetic Composition Containing Hairy Rat Extract, Rhododendron Extract and Painted Wood Extract

Formulation Examples 1 to 16 were prepared as the following cosmetic preparations containing the extract of Preparation Example according to the present invention as shown in Tables 9 and 10 below. Comparative Examples 1 to 3, which each contain a pine needle extract, Rhodiola extract and Concha tree extract, and comparative formulation examples 4 and 5, which contain glycerin and 1,3-butylene glycol as a moisturizing agent instead of the extract, Comparative Formulation Example 5 containing adenosine as the wrinkle improving material instead of the extract, Comparative Formulation Example 6 containing no extract or moisturizing agent, Comparative Formulation Example 7 containing the niacinamide as the whitening material instead of the extract, .

division ingredient Formulation Example 1 Formulation Example 2 Formulation Example 3 Formulation Example 4 Formulation Example 5 Formulation Example 6 A Production Example 7 2 - - - - - Production Example 9 - 2 - - - - Production Example 10 - - 2 - - - Production Example 12 - - - 2 - - Production Example 13 - - - - 2 - Production Example 15 - - - - - 2 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 to 100 B Cetostearyl alcohol 2 2 2 2 2 2 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 Microcrystalline 0.7 0.7 0.7 0.7 0.7 0.7 Squalane 5 5 5 5 5 5 Liquid paraffin 3 3 3 3 3 3 Trioctanoin 5 5 5 5 5 5 Polysorbate 1.2 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3 3 3 3 3 3 BHT 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

division ingredient Formulation Example 7 Formulation Example 8 Formulation Example 9 Formulation Example 10 Formulation Example 11 Formulation Example 12 Formulation Example 13 Formulation Example 14 Formulation Example 15 Formulation Example 16 A Production Example 19 2 - - - - - - - - - Production example 20 - 2 - - - - - - - - Production Example 21 - - 2 - - - - - - - Production Example 22 - - - 2 - - - - - - Production Example 23 - - - - 2 - - - - - Production Example 26 - - - - - 2 - - - - Production Example 27 - - - - - - 2 - - - Production Example 28 - - - - - - - 2 - - Production Example 29 - - - - - - - - 2 - Production Example 30 - - - - - - - - - 2 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 Cetostearyl alcohol 2 2 2 2 2 2 2 2 2 2 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 B Microcrystalline 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7 Squalane 5 5 5 5 5 5 5 5 5 5 Liquid paraffin 3 3 3 3 3 3 3 3 3 3 Trioctanoin 5 5 5 5 5 5 5 5 5 5 Polysorbate 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3 3 3 3 3 3 3 3 3 3 BHT 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

division ingredient Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Comparative Formulation Example 4 Comparative Formulation Example 5 Comparative Formulation Example 6 Comparative Formulation Example 7 A Production Example 1 2 - - - - - - Production Example 2 - 2 - - - - - Production Example 3 - - 2 - - - - glycerin - - - 5 - - - 1,3-butylene glycol - - - 5 - - - Adenosine - - - - 3 - - Niacinamide - - - - - - 2 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 to 100 to 100 Cetostearyl alcohol 2 2 2 2 2 2 2 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 B Microcrystalline 0.7 0.7 0.7 0.7 0.7 0.7 0.7 Squalane 5 5 5 5 5 5 5 Liquid paraffin 3 3 3 3 3 3 3 Trioctanoin 5 5 5 5 5 5 5 Polysorbate 1.2 1.2 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3 3 3 3 3 3 3 BHT 0.05 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

Experimental Example  10: Fur extract, Rhodiola extract  And Painted Tree Extract Skin barrier function  Improvement effect

The skin barrier function improving effects of Formulation Examples 1 to 16 and the comparative formulations were evaluated. A total of 210 adult males and females over 20 years old were randomly divided into 21 groups (1 to 21 groups), followed by a 20-minute fin-chamber application of 1.0% SLS aqueous solution to a total of 210 females. . Thereafter, Formulation Examples 1 to 16 and Comparative Formulation Examples 1 to 4 and 6 were applied to Groups 1 to 19, respectively, twice a day for one week. The effect of improving the skin barrier function was measured by Transepidermal Water Loss (TEWL) for 7 days after SLS application, coaptation, and application of the formulation. The results are shown in Table 12.

Before the test SLS Collaboration Day1 Day2 Day3 Day4 Day5 Day6 Day7 Formulation Example 1 6.5 21.5 18.9 16.2 12.5 10.5 7.2 6.5 6.5 Formulation Example 2 6.7 21.3 19.5 16.7 12.4 10.3 7.6 6.5 6.4 Formulation Example 3 6.7 22.4 20.4 15.4 12.6 11.1 7.5 6.4 6.5 Formulation Example 4 6.6 22.3 19.8 16.1 12.5 9.8 7.4 6.4 6.5 Formulation Example 5 6.6 21.6 19.2 16 12.3 9.9 7.7 6.5 6.4 Formulation Example 6 6.7 20.5 19.3 15.5 12.6 10.2 7.6 6.4 6.5 Formulation Example 7 6.5 22.3 19.3 9.7 7.3 6.8 6.7 6.5 6.4 Formulation Example 8 6.7 22.1 19.1 9.8 7.5 6.9 6.5 6.6 6.5 Formulation Example 9 6.8 22.3 18.8 10.3 7.7 6.7 6.5 6.5 6.5 Formulation Example 10 6.7 21.8 19.1 9.7 7.6 6.7 6.8 6.6 6.7 Formulation Example 11 6.5 21.7 18.8 9.5 7 6.8 6.7 6.7 6.5 Formulation Example 12 6.8 21.5 18.9 9.3 7.3 6.8 6.7 6.6 6.7 Formulation Example 13 6.5 22 19.2 10.1 7.2 6.7 6.6 6.7 6.5 Formulation Example 14 6.7 21.4 19.4 10.1 7.4 6.8 6.5 6.7 6.8 Formulation Example 15 6.8 22 19.1 9.8 7.3 6.7 6.6 6.7 6.7 Formulation Example 16 6.7 21.7 15.8 10.3 7.4 6.8 6.5 6.8 6.7 Comparative Formulation Example 1 6.7 21.4 20.4 18.1 14.4 11.2 8.7 7.2 6.8 Comparative Formulation Example 2 6.8 22 20.1 17.8 14.3 11.7 8.6 7.3 6.7 Comparative Formulation Example 3 6.7 21.7 18.8 18.3 14.4 11.3 8.5 7.1 6.7 Comparative Formulation Example 4 6.5 21.5 17.6 13.2 8.5 6.9 6.5 6.4 6.6 Comparative Formulation Example 6 6.5 22.1 20.2 19.3 18.5 17.3 15.7 12.6 10.4

As can be seen from Table 12, it was confirmed that the areas coated with Formulation Examples 1 to 16 restored the barrier function more rapidly than Comparative Formulation Examples 4 and 6. [ In particular, it was confirmed that Formulation Examples 7 to 16 containing a mixture of three kinds of extracts recovered the barrier function more rapidly.

Experimental Example 11: Moisturizing effect of the extracts of Walnut, Rhodochrosite and Painted Wood

In order to measure the moisturizing effect of the cosmetic compositions of Formulation Examples 1 to 16, clinical tests were carried out as follows. A total of 210 healthy adult men and women who felt dry skin were divided into 21 groups (1 to 21 groups) by 10 persons, and the cosmetic compositions of Formulation Examples 1 to 16 were administered to groups 1 to 16 at 17 to 19 (Comparative Formulation Example 4) containing glycerin and 1,3-butylene glycol as moisturizing agents instead of extracts (Comparative Formulation Example 4), Comparative Example Formulations 1 to 3 containing cosmetic compositions containing no extracts and moisturizers The composition (Comparative Formulation Example 6) was applied to the face for 2 weeks each 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 13 below.

division Before use After 2 weeks of use After 4 weeks of use Formulation Example 1 22.4 31.9 35.64 Formulation Example 2 22.3 32.5 35.7 Formulation Example 3 21.6 31.4 35.4 Formulation Example 4 21.8 32.7 36.1 Formulation Example 5 22.3 32.5 36.3 Formulation Example 6 22.1 33.5 35.7 Formulation Example 7 21.7 40.1 55.9 Formulation Example 8 22.0 40.3 56.1 Formulation Example 9 23.1 38.6 55.7 Formulation Example 10 23.4 39.4 55.8 Formulation Example 11 22.8 39.3 56.2 Formulation Example 12 23.2 40.5 54.7 Formulation Example 13 23.5 40.7 55.1 Formulation Example 14 21.4 38.9 52.3 Formulation Example 15 23.5 39.8 55.8 Formulation Example 16 22.9 40.7 56.9 Comparative Formulation Example 1 21.4 28.9 31.3 Comparative Formulation Example 2 23.5 29.8 30.8 Comparative Formulation Example 3 22.9 27.7 30.9 Comparative Formulation Example 4 23.3 37.4 49.7 Comparative Formulation Example 6 22.2 24.8 23.8

As shown in Table 13, it was confirmed that the moisturizing effect of the cosmetic compositions containing the extract of the present invention (Formulation Examples 1 to 16) was superior to the cosmetic compositions containing other moisturizing agents (Comparative Formulation Example 4). Especially, it was confirmed that Formulation Examples 7 to 16 containing a mixture of three kinds of extracts had better moisturizing effect.

EXPERIMENTAL EXAMPLE 12: Whitening effect of extracts of Wooly Rhododendrons, Rhodophilus edodes, and Painted Woods

Clinical trials for skin whitening effects of cosmetic compositions containing the extract of the present invention were conducted. Clinical trials were evaluated through actual use tests of each cosmetic product. Twenty-one groups (1 to 21 groups) of 10 women (30 to 50 years old) were randomly divided into 10 groups, and the cosmetic compositions of Formulation Examples 1 to 16 were administered to Groups 1 to 16, Comparative Formulation Examples 1 to 3 were applied to a cosmetic composition (Comparative Formulation Example 4) containing glycerin and 1,3-butylene glycol as a moisturizing agent instead of an extract in Group 20, a cosmetic composition containing 2% by weight of niacinamide in Group 21 (Comparative Formulation Example 7) were applied to the face for 2 weeks each for 4 weeks. The brightness (L *) was measured with a Chromameter (Minolta CR300) using both sides of the face, and the whitening effect after 4 weeks was confirmed. The results of the whitening effect based on this evaluation are shown in Table 14 below.

division The measured value (L *) Before use After 2 weeks of use After 4 weeks of use Formulation Example 1 58.5 61.4 61.8 Formulation Example 2 58.5 61.5 62.9 Formulation Example 3 58.5 61.3 61.8 Formulation Example 4 58.4 61.8 61.9 Formulation Example 5 58.1 60.5 61.7 Formulation Example 6 57.8 60.1 61.2 Formulation Example 7 58.1 62.0 65.2 Formulation Example 8 58.3 62.7 65.9 Formulation Example 9 58.2 63.2 65.8 Formulation Example 10 57.9 62.3 65.1 Formulation Example 11 57.8 63.4 66.0 Formulation Example 12 58.1 63.5 66.3 Formulation Example 13 58 63.1 66.1 Formulation Example 14 58.6 62.2 66.2 Formulation Example 15 58.7 63.8 65.9 Formulation Example 16 58.6 62.5 65.2 Comparative Formulation Example 1 58.6 59.2 60.1 Comparative Formulation Example 2 58.7 59.8 60.5 Comparative Formulation Example 3 58.6 60.5 60.2 Comparative Formulation Example 4 58.1 58.5 58 Comparative Formulation Example 7 58.2 61.4 64.9

As shown in Table 14, it was confirmed that the whitening effect of the cosmetic compositions containing the extract of the present invention (Formulation Examples 1 to 16) was superior to the cosmetic containing the whitening component (Comparative Formulation Example 7). Especially, it was confirmed that the whitening effect of Formulation Examples 7 to 16 containing a mixture of three kinds of extracts was more excellent.

Experimental Example  13: Fur extract, Rhodiola extract  And skin tree extracts

Experimental Example 13 was conducted to evaluate the skin elasticity improving effect of Formulation Examples 1 to 16, Comparative Formulation Examples 1 to 3 containing the extract of the present invention and Comparative Formulation Example 4 containing a moisturizer instead of the extract, 200 women with reduced strength in their 40s were measured. 10 groups were divided into 20 groups, and Formulations 1 to 16 were applied to Groups 1 to 16, Comparative Formulations 1 to 3 were prepared for Groups 17 to 19, and Comparative Formulation 4 was prepared for Group 20, 2 times for 12 weeks. After 8 weeks, skin elasticity was measured. 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 15 below, and the results are an average value of improvement for each group.

Experimental product Skin elasticity improvement (%) Formulation Example 1 18.5 Formulation Example 2 19.1 Formulation Example 3 19.4 Formulation Example 4 18.2 Formulation Example 5 19.5 Formulation Example 6 18.3 Formulation Example 7 32.2 Formulation Example 8 31.9 Formulation Example 9 32 Formulation Example 10 32.1 Formulation Example 11 31.4 Formulation Example 12 32.1 Formulation Example 13 31.5 Formulation Example 14 31.8 Formulation Example 15 32.4 Formulation Example 16 31.2 Comparative Formulation Example 1 15.8 Comparative Formulation Example 2 13.4 Comparative Formulation Example 3 14.2 Comparative Formulation Example 4 6.3

As shown in Table 15, Formulations 1 to 16 containing the extracts showed significantly improved skin elasticity as compared to Comparative Formulations 1 to 4. Especially, it was confirmed that Formulation Examples 7 to 16 containing a mixture of three kinds of extracts were more excellent in improving the elasticity.

Experimental Example  14: Fur extract, Rhodiola extract  And wrinkle of skin extract

Clinical trials were carried out on the effect of the cosmetic composition containing the extract of the present invention to improve the skin wrinkles. Clinical trials were evaluated through actual use tests of each cosmetic product. A total of 210 women (30 to 50 years old) were randomly divided into 21 groups of 10 individuals each, and Formulation Examples 1 to 16 were applied to Groups 1 to 16, Comparative Formulation Examples 1 to 5 Was applied twice a day for 8 weeks, and the wrinkle improvement effect after 8 weeks using both sides of the face was evaluated by visual observation and instrument measurement (cutometer SEM 575, C + K Electronic Co., Germany) As shown in Table 16 below.

Wrinkle improvement effect Effective rate (%) Great slightly none Formulation Example 1 7 5 8 60 Formulation Example 2 7 6 7 65 Formulation Example 3 6 7 7 65 Formulation Example 4 5 7 8 60 Formulation Example 5 5 7 8 60 Formulation Example 6 5 8 7 65 Formulation Example 7 16 4 One 95 Formulation Example 8 15 5 2 90 Formulation Example 9 15 5 2 90 Formulation Example 10 16 4 One 95 Formulation Example 11 17 3 One 95 Formulation Example 12 17 3 One 95 Formulation Example 13 13 7 2 90 Formulation Example 14 10 8 2 90 Formulation Example 15 11 7 2 90 Formulation Example 16 13 6 One 95 Comparative Formulation Example 1 4 4 12 40 Comparative Formulation Example 2 3 5 12 40 Comparative Formulation Example 3 3 5 12 40 Comparative Formulation Example 4 3 3 14 30 Comparative Formulation Example 5 12 6 3 85

As can be seen from the results of Table 16, the cosmetic compositions of Formulation Examples 1 to 16 containing the extract of the present invention showed a wrinkle-improving effect about 3.0 times higher than Comparative Formulation Example 4 containing a moisturizer. Especially, it was confirmed that the wrinkle improving effect of Formulations 7 to 16 containing a mixture of three kinds of extracts was more excellent. Similar results were obtained with the cosmetic preparation of Comparative Formulation Example 5 containing adenosine, which is known to have a wrinkle-reducing effect instead of the extract.

Experimental Example  15: Wool crumb extract, goldthread  Extract and Painted Tree Extract Dermal density  Measure improvement

Clinical trials were conducted on the effect of improving the dermal density of cosmetic preparations containing the extract of the present invention. Clinical trials were evaluated through actual use tests of each cosmetic product. 200 women (30 to 50 years old) were divided into 20 groups of 10 individuals each, and Formulation Examples 1 to 16 were applied to Groups 1 to 16, Comparative Formulation Examples 1 to 5 were applied to groups 17 to 20 After applying to the face twice a day for 4 weeks, the improvement of the dermatome density after 4 weeks using both sides of the face was confirmed by a device evaluation using SkinScanner DUBcutis (tpm (terbana pro medicum), Luneburg, Germany). The dermal density increase rate was calculated according to the following equation (4), and the dermal density improvement effect is shown in Table 17 below.

Experimental product Density increase rate of dermis (%) Formulation Example 1 16.7 Formulation Example 2 16.8 Formulation Example 3 17.7 Formulation Example 4 17.3 Formulation Example 5 17.9 Formulation Example 6 18.1 Formulation Example 7 26.7 Formulation Example 8 26 Formulation Example 9 27.7 Formulation Example 10 27.1 Formulation Example 11 26.5 Formulation Example 12 26.2 Formulation Example 13 25.6 Formulation Example 14 27.1 Formulation Example 15 26.7 Formulation Example 16 27.7 Comparative Formulation Example 1 10.8 Comparative Formulation Example 2 11.1 Comparative Formulation Example 3 10.7 Comparative Formulation Example 4 3.5 Comparative Formulation Example 5 25.1

As shown in Table 16, it can be seen that Formulations 1 to 16 containing the extract significantly improved skin dermal density as compared to Comparative Formulations 1 to 5. In particular, it was confirmed that Formulation Examples 7 to 16 containing the mixture of the three kinds of extracts were more excellent in improving the dermal density.

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

Claims (4)

Wherein the plant extract comprises two kinds of plant extracts selected from the group consisting of pine root extract,
The weight ratio of the two plant extracts was 1: 1,
The plant extract is extracted by fermentation from microorganisms or extraction using natural fermentation metabolism,
A skin elasticity improvement effect, a skin wrinkle improvement effect and a dermis densification improvement effect. The method according to claim 1,
Wherein the plant extract is contained in an amount of 0.001 to 30.0% by weight based on the total weight of the cosmetic composition. The method according to claim 1,
Wherein the microorganism is Lactobacillus plantarum ATCC 10012. delete

Images