WO2007083868A1

WO2007083868A1 - Cosmetic composition containing extracts of pinus koraiensis as active ingredient

Info

Publication number: WO2007083868A1
Application number: PCT/KR2006/002549
Authority: WO
Inventors: Han Sung Kim; Youn Joon Kim; Sung Il Lee; Hee Kyung Joo; Duck Hee Kim; Han Kon Kim; Ih Seop Chang
Original assignee: Amorepacific Corporation
Priority date: 2006-01-19
Filing date: 2006-06-30
Publication date: 2007-07-26
Also published as: JP5175744B2; CN101360478B; KR101252548B1; JP2009523785A; CN101360478A; KR20070076656A

Abstract

The present invention relates to a cosmetic composition comprising extract of Pinus koraiensis and at least one selected from the group consisting ofextract of Primus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient. More particularly, the cosmetic composition according to the present invention comprises extract of Pinus koraiensis and at least one selected from the group consisting ofextract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient to show the effect promoting the growth of keratinocyte, the effect eliminating oxygen free radicals, the effect promoting the biosynthesis of Type 1 procollagen by reduction of the biosynthesis of MMP-I , the effect preventing aging and the effect moisturizing skin.

Description

[DESCRIPTION]

[Invention Title]

Cosmetic composition containing extracts of Pinus koraiensis as active ingredient [Technical Field]

The present invention relates to a cosmetic composition comprising extract of Pinus koraiensis and at least one selected from the group consisting ofextract of Prunus mume, extract of Phaseolus angularis, extract of

Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient. More particularly, the cosmetic composition according to the present invention comprises extract of Pinus koraiensis and at least one selected from the group consisting ofextract of Prunus mume, extract of

Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient, to show the effect promoting the growth of keratinocyte, the effect removing oxygen free radicals, the effect promoting biosynthesis of Type 1 procollagen by reduction of biosynthesis of MMP-I, the effect preventing skin aging and the effect moisturizing skin.

[Background Art]

The skin which is a primary defensive membrane to protect organs in the body from change of temperature and humidity and external environment such as UV rays, pollutants and the like and to play an important role in maintaining homeostasis such as body temperature control. However, UV rays, excessive physical and chemical stimulation from the outside, stress and malnutrition may obstruct normal functions of the skin and urge skin aging phenomena such as elasticity loss, cornification, wrinkle formation and the like.

Particularly, as new keratinocytes are produced orderly and continuously and old keratinocytes in the outermost layer occasionally come off, if the cornification to maintain the outer skin to a regular thickness does not proceed normally, keratinocytes are abnormally produced and accumulated at the surface of the skin to increase the skin thickness, roughen and darken the skin and form fine wrinkles, thereby causing skin aging. Also, production of oxygen free radicals by UV rays and reduction of biosynthesis of Type 1 procollagen through biosynthesis of MMP-I by UV rays associated with decomposition of the skin substrate relate to the skin aging.

Therefore, in order to prevent the above-described phenomena and maintain healthier and more elastic skin, it has been examined the use of cosmetic products supplemented with bioactive materials obtained from various animals, plants and microorganisms to maintain original functions of the skin and activate dermal cells, thereby effectively inhibiting the skin aging. However, conventional materials of cosmetic products mostly have problems in insufficient effects and induction of side effects. [Disclosure] [Technical problem]

Therefore, the present inventors have made efforts to find natural materials having the effect moisturizing skin and the effect preventing the skin aging without causing side effects on skin moisturizing and found that a composition comprising extract of Pinus koraiensis and at least one selected from the group consisting ofextract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient has the effect promoting the growth of keratinocyte, the effect removing oxygen free radicals, the effect promoting biosynthesis of Type 1 procollagen by reduction of biosynthesis of MMP-I, the effect preventing aging and the effect moisturizing skin. Based on the above finding, the present invention has been completed.

Therefore, the object of the present invention is to provide a cosmetic composition comprising extract of Pinus koraiensis and at least one selected from the group consisting extract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient to show the effect promoting the growth of keratinocyte, the effect removing oxygen free radicals, the effect promoting biosynthesis of Type 1 procollagen by reduction of biosynthesis of MMP-I, the effect preventing aging and the effect moisturizing skin. [Technical solution] In order to accomplish the above object, according to the present invention, there is provided a cosmetic composition comprising extract of Pinus koraiensis and at least one selected from the group consisting of extract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient. The cosmetic composition according to the present invention shows the effect promoting the growth of keratinocyte, the effect removing oxygen free radicals, the effect promoting biosynthesis of Type 1 procollagen by reduction of biosynthesis of MMP-I, the effect preventing aging and the effect moisturizing skin. Now, the respective active ingredients of the cosmetic composition according to the present invention are described in detail. Pinus koraiensis is dried ripen seed of Korean white pine belonging to the Family Pinaceae and its nut is called "Baekja", "Songja" and "Silback" as a herb medicine, which is contained in a cone looking like that of Korean pine tree and has a shape of a triangular egg. From old times, Pinus koraiensis has been generally used as a nutritive tonic and known to be good in the period of convalescence. Also, it is a high calorie food rich in fats and proteins, particularly rich in vitamin B, and functions to make the skin smooth and to lower the blood pressure.

Prunus mume is fruit of Japanese apricot tree belonging to the Family Rosaceae and generally called "Maesil". It is recorded in Shinnongbonchogyeong, a Chinese ancient book, that Prunus mume is one of the oldest fruit tree used for medicinal purpose. In various books on medicine such as Dongeui Bogam (Thesaurus of Eastern Medicine), there are records on the use of Prunus mume as herb medicines. In herb medicines, Prunus mume is used to show the effect making the stomach strong, stopping bleeding, stopping diarrhea, discharging phlegm, discharge alcoholic poison and neutralizing poison. Prunus mume is a alkali food and good for recovery from fatigue and has effects of improving one's physical constitution. Particularly, it is excellent in neutralizing poison and good for skin and has antibacterial effect. Recently, its potential for use as an anti-cancer food has been studied.

Phaseolus angularis is a plant belonging to the Family Leguminosae and Sodoo is a herb medicine name for red bean. Its origin is assumed to be a region of China and cultivated in the oriental region from the ancient. In herb medicine, it is used in controlling dropsy, treating blood extravasation, accumulating energy, treating swelling, removing wetness in the body, promoting blood circulation, neutralizing poison, discharging purulence, strengthening the spleen and the stomach, strengthening mulscle and bone .

Chaenomeles sinensis is a fruit of Chaenomeles sinensis tree which is a cultivated plant native to china. It looks like melon but hangs on the tree. Therefore, it is also called tree fruit. Main components of Chaenomeles sinensis include saponin, organic acid, flavonoid, tannin and the like. In herb medicine, it is used in releasing abnormal stain condition of muscle, vitalizing blood vessels, leveling the energy of the liver, treating functional disorder of the stomach, removing wetness in the body and alleviating pain. Sesamum indicum is known to be effective in chronic gastroenteritis, neuronitis, high blood pressure, constipation, robustness, anemia and the like and acts to reduce cholesterol, facilitate blood circulation and make skin or hairs gloss. In the herb medicine, it is used in making five organisms in good condition, strengthening the liver and the kidney, strengthening hairs, strengthening functions of the eyes and ears, growing fat and strengthening functions of the intestines. The proteins of Sesamum indicum are mainly globulin and also contain y -tocopherol to help antioxidization. Therefore, it can prevent skin aging by the anti-oxidizing action and vitalizing skin metabolism to prevent pigmentation in a small area which turns black. The extracts included in the cosmetic composition according to the present invention are prepared by a method known to the persons skilled in the art and commonly used to extract an active ingredient from plants. For example, a pulverized herb medicine is mixed with water, anhydrous or hydrated low alcohol having 1 to 3 carbon atoms, acetone, ethyl acetate or diethyl ether in an amount of 1 to 15 times volume based on the dry weight of the pulverized, extracted in an extractor, equipped with a cooling condenser, at a temperature of 50 to 100 °C for 5 to 24 hours while heating; mixed with water, anhydrous or hydrated low alcohol having 1 to 3 carbon atoms, acetone, ethyl acetate or diethyl ether in an amount of 1 to 15 times volume based on the dry weight of the pulverized, extracted by dipping at a temperature of 4 to 25 ^°C for 3 to 20 days 5 to 24 hours. Alternatively, the anhydrous or hydrated ethanol or methanol extract is mixed with a proper amount of water, followed by filtration to remove precipitations. The filtrate is thoroughly mixed with ethyl acetate, butanol or diethyl ether, left for a while for phase separation. The upper layer is separated and condensed in an extractor equipped with a cooling condenser to obtain an extract. The filtrate of the herb extract obtained by the above-described method is concentrated in vacuum, dried and combined into a cosmetic composition.

The extracts according to the present invention may be used in a mixture of the extracts which are separately extracted from the herbs or in an extract which is extracted from a mixture of the herbs.

The cosmetic composition comprises, as an active ingredient, the extract of Pinus koraiensis and at least one selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum and the active ingredient is in an amount of 0.0001 to 25 wt%, based on the total weight of the composition. If the active ingredient is less than 0.0001 wt% the effect is insufficient. If it exceeds 25 wt%, there may be problems in the use feeling of the product and stability of the formulation.

The cosmetic composition according to the present invention shows the effect promoting the growth of keratinocyte, the effect eliminating oxygen free radicals, the effect promoting the biosynthesis of Type 1 procollagen by reduction of the biosynthesis of MMP-I, the effect preventing aging and the effect moisturizing skin.

Also, in addition to the extract according to the present invention, the cosmetic composition may comprise other ingredients which may provide synergic effect to the main effect the present invention within the scope in which the main effect the present invention is not impaired. The additive ingredients may be properly selected and combined by a person in the art according to the formulation and purpose of the cosmetic composition.

For example, the cosmetic composition may comprise a skin absorption promoter in order to increase the effect. Also, the cosmetic composition may further comprise one selected from the group consisting of water-soluble vitamin, oil-soluble vitamin, polypeptide, polysaccharide, sphingolipid and sea alga extracts.

Moreover, the cosmetic composition according to the present invention may further comprise an additive, which is commonly added to a cosmetic composition, as needed, in addition to the essential ingredients.

Examples of the additive includes, oily components, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powder, UV ray absorbent, preservative, anti-bacteria, antioxidants, plant extracts, pH controller, alcohol, colorants, fragrance, blood circulation promoter, cooling agent, anti-perspiration, purified water and the like.

The additives which may be added to the cosmetic composition according to the present invention are not limited to the above-described.

Also, the mixing ratio of the additives is preferably 0.01 to 10 wt%, more preferably 0.01 to 10 wt%, though it is not limited to the foregoing range but may be vary within the scope in which the purpose and the effect the present invention are not impaired.

The cosmetic composition according to the present invention may be formulated without any particular limit. For example, the formulation includes milk lotion, cream, skin lotion, essence, pack, gel, powder, lip-stick, makeup base, foundation, lotion, ointment, patch, beauty liquid, cleansing foam, cleansing cream, cleansing water, soap or spray. [Best Mode]

Hereinafter, the present invention is further described in detail through Examples and Experimental Examples. However, the present invention is not limited thereto .

[Reference 1] Preparation of extract of herb medicines Each of Pinus koraiensis, Prunus mume, Phaseolus angularis, Chaenomeles sinensis and Sesamum indicum (Kunwha Pharmaceutical Company Ltd.) was collected and high quality herbs were selected, washed to remove impurities and dried in the shade. Then, 100 g of each of dried Pinus koraiensis, Prunus mume, Phaseolus angularis, Chaenomeles sinensis and Sesamum indicum was added to 2 L of purified water and extracted in a reflux apparatus at about 95 to 100^°C for 18 hours while heating. After the completion of the extraction, the extract was successively distilled and concentrated to the total weight of about 500 g and cooled to room temperature. Then, the product was put into a filter cloth, pressed to separate and remove the solid content and filtered to obtain viscous concentrate. The concentrate was vacuum concentrated at 80 ^"C to remove water completely, dried in vacuum to obtain 5.0 g of Pinus koraiensis, 1O g of Prunus mume, 5.0 g of Phaseolus angularis, 5.0 g of Chaenomeles sinensis and 5.0 g of Sesamum indicum as dried powder.

[Examples 1 to 15 and Comparative Example 1] Each herb extract obtained by extraction with water in Reference 1 was mixed according to description in Table 1 and used in experiments of Examples 1 to 15 and Comparative Example 1. Here, the mixing ratio of each herb extract was the same. [Table 1 ]

[Experimental Example 1] Effect promoting growth of keratinocyte

(MTT assay)

The effect of the extracts of Examples 1 to 15 and Comparative Example 1 on growth of keratinocyte was measured by the MTT method and the method was briefly described as follows. Firstly, keratinocytes were seeded in a 96 well plate in a concentration of 5 X 10³ /200 μi/wel\, cultured for 24 hours and treated with the extracts of Examples 1 to 15 and Comparative Example 1 at a concentration of 0.0001, 0.001, 0.01 and 0.1 %, cultured for 48 hours. After, the culture fluid was removed by suction, the cells were washed once with PBS, treated with 100 μi of MTT (Methylthiazolyldiphenyl-tetrazolium bromide, Sigma, USA) solution (0.5 mg/m#) and cultured for 4 hours at 37 ⁰C , 5% CO₂. After the culture fluid was removed by suction, the cells were treated with 200 μl of DMSO (dimethyl sulfoxide) solution, stirred in a shaker for 10 minutes and measured for absorption at 540 run on ELISA reader (DIbiotech, Korea. Here, the non-treatment group which had not been treated with the extracts of Examples 1 to 15 and Comparative Example 1 was used as control. The results are shown in Table 2. [Table 2]

As shown in Table 2, it was noted that the effects were shown in all the cases and generally, when the concentration of the used extract was increased the effect promoting the growth of keratinocytes was greater. Also, the compositions of Examples 1 to 15 including at least one selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum in addition to the extract of Pinus koraiensis were more effective than the composition of Comparative Example 1 including only the extract of Pinus koraiensis and the kinds of the extracts selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum added to the extract of Pinus koraiensis was increased, the effect promoting the growth of keratinocytes was greater.

[Experimental Example 2] Effect eliminating oxygen free radicals In order to examine the effect of the extracts of Examples 1 to 15 and Comparative Example 1 inhibiting production of oxygen free radicals produced by UV rays, that is, the effect eliminating oxygen free radicals, the following experiment was performed using a fluorescent material.

The cell line used in the experiment was the human keratinocytes HaCaT cell line distributed by Dr. Fusenig in The German Cancer Research Center. The cells were plated in a 96 well black plate for fluorescence measurement in an amount of 2.0 X 10⁴ per well, cultured in DMEM (Dulbeccos Modification of Eagles Medium, FBS 10%, Gibco, USA) supplemented with penicillin/streptomycin at 37 ^°C , 5% CO₂ for 1 day and treated with the extracts of Examples 1 to 15 and Comparative Example 1 at a concentration of 0.5, 0.25, 0.125, 0.0625 and 0.03125 %.

Then, the test samples were added to the wells and cultured for 24 hours. The plate was washed with HCSS (HEPES-buffered control salt solution, Gibco, USA) to remove the remaining medium. 100 μi of 20 μ M DCFH-DA (2',7^l-dichlorodihydro-fluorescein diacetate, Molecular Probes, USA) was added to the HCSS, cultured at 37 ^°C , 5 % CO₂ for 20 minutes and washed with HCSS. Then, the fluorescence of DCF (dichlorofluorescein) oxidized to oxygen free radicals after 100 βi of HCSS which had been treated with the sample at different concentrations had been added was measured on fluorescence plate reader (Ex=485 nm, Em=530 nm). UVB(30mJ/cm²) was irradiated and the fluorescence right after the irradiation (Table 3) and after 3 hours (Table 4) were measured on fluorescence plate reader (Ex=485 nm, Em=530nm). Here, as control, the non-treatment group which had not been treated with the extracts of Examples 1 to 15 and Comparative Example 1 was used. [Table 3]

[Table 4]

As shown in Table 3 and Table 4, it was noted that all the test samples showed the effect inhibiting oxygen free radicals and when the concentration of the extracts was increased and the production of oxygen free radicals were generally more inhibited. Also, the compositions of Examples 1 to 15 including at least one selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum in addition to the extract of Pinus koraiensis were more excellent than the composition of Comparative Example 1 including only the extract of Pinus koraiensis and when the kinds of the extracts selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum added to the extract of Pinus koraiensis was increased, the production of oxygen free radicals were more inhibited. Also, the effect was greater 3 hours later than right after the treatment.

[Experimental Example 3] MMP-I and procollagen assay In order to examine the effect of the extracts of Examples 1 to 15 and Comparative Example 1 promoting biosynthesis of Type 1 procollagen by reducing biosynthesis of MMP-I by UV rays associated with decomposition of skin substrates, the MMP-I and procollagen assay was performed as follows.

The normal epidermis with the fat layer removed was chopped into small pieces to separate the epidermis and the dermis by collagenase. The epidermis and the dermis tissues were separately taken into 0.25 % trypsin solution and incubated at 37 ^°C , 5% CO₂ for 10 minutes. Then, the vortex was performed to isolate keratinocytes and fibroblasts. The isolated cells were collected and washed. The keratinocytes were cultured in KGM (keratinocyte growth medium, Clonetics, USA) at a concentration of 1 X lO⁴ cells/cm² and the fibroblasts were cultured in DMEM supplemented with 10 % fetal bovine serum (FBS). When the cells grew 70 to 80 %, they were distributed in a mixing ratio of 1:3 and subcultured. The third and fourth subcultured cells were used in the experiment.

In order to measure the amount of MMP-I, the fibroblasts were cultured in a 48 well plate to at least 90 % and left under starvation condition for one day. Then, the cells were washed two times with PBS, treated with 100 μi of PBS and irradiated by UV ray A (Dermlight cube 401 with UVA filters mounted, UVAtec, USA) at A 15J/cm² while the cover of the plate was opened. After the irradiation, the cells were washed once with PBS, placed in fetal bovine serum free DMEM containing the extracts of Examples 1 to 15 and Comparative Example 1 in a portion of 0.001 and 0.01 % and incubated at 37 ^°C , 5% CO₂ for 48 hours. The amount of MMP-I isolated in the medium was measured using the human ELISA system (RPN2610, Amersham Pharmacia Biothch, UK) and corrected by the total protein amount of the fibroblast (Table 5). Here, as control, the non-treatment group which had not been treated with the extracts of Examples 1 to 15 and Comparative Example 1 was used.

Meanwhile, in order to measure the amount of procollagen, the cells under the same conditions to measure the amount of MMP-I were treated with the extracts of Examples 1 to 15 and Comparative Example 1 in a portion of 0.001 and 0.01% at the starvation condition without UV irradiation. After 24 hours, the amount of procollagen isolated in the medium was measured using the procollagen type-1 C-peptide EIA kit (MKlOl, Takara, Japan) (Table 6). As control, the non-treatment group which had not been treated with the extracts of Examples 1 to 15 and Comparative Example 1 was used. [Table 5]

[Table 6]

As shown in the effect inhibiting the biosynthesis of MMP-I of Table 5, it was noted that the effect was shown in all the cases and generally, when the concentration of the used extract was increased the amount of the produced MMP-I was reduced. Also, the compositions of Examples 1 to 15 including at least one selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum in addition to the extract of Pinus koraiensis were more effective than the composition of Comparative Example 1 including only the extract of Pinus koraiensis and the the kinds of the extracts selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum added to the extract of Pinus koraiensis was increased, the amount of the produced MMP-I was reduced.

Also, as shown in the effect increasing the biosynthesis of Type 1 procollagen of Table 6, it was noted that the effect was shown in all the cases and generally, when the concentration of the used extract was increased the amount of the produced procollagen was increased. Also, the compositions of Examples 1 to 15 including at least one selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum in addition to the extract of Pinus koraiensis were more effective than the composition of Comparative Example 1 including only the extract of Pinus koraiensis and the the kinds of the extracts selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum added to the extract of Pinus koraiensis was increased, the amount of the produced procollagen was increased.

[Formulation 1 to 15 and Comparative Formulation 1] Preparation of cosmetic composition The cosmetic compositions of Formulations 1 to 15 and Comparative

Formulation 1 were prepared using the extracts of Examples 1 to 15 and Comparative Example 1 according to the composition described in Table 7. The oily phase of the Ingredient Nos. 1 to 7 and the aqueous phase of the Ingredient Nos. 8 to 15 in the composition of Table 7 were separately dissolved by heating. The oily phase was slowly added to the aqueous phase, while stirring, for emulsification.

The cosmetic compositions of Formulations 1 to 15 comprised, as an active ingredient, the extract of Pinus koraiensis and at least one selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum while the cosmetic composition of Comparative Formulation 1 included only the extract of Pinus koraiensis. The oily phase and the aqueous phase of the compositions were dissolved completely at 65 to 75 ^°C and emulsified at 7,000 to 8,000 rpm for 5 minutes to prepare a cream formulation.

[Table 7]

[Experimental Example 4] Effect improving wrinkles In order to examine the effect improving wrinkles of Formulation 15 and Comparative Formulation 1 , the following process was performed.

140 women of thirties were divided into 7 groups of 20 persons and had cream formulations of Formulation 15 and Comparative Formulation 1 applied around eyes 2 times per day for 8 weeks. The wrinkle condition was measured using a replica made of silicone on a visiometer (SV60, Courage+Khazaka electronic GmbH, Germany) as a image analyzer. As control, the non-treatment group which had not been treated with creams of Formulation 15 and Comparative Formulation 1 was used. The result which is an average of values obtained by subtracting a parameter after 8 weeks from a parameter before 8 weeks is shown in Table 8. That is, the negative value means that the effect improving wrinkles is greater. [Table 8]

From the result of Table 8, it was noted that the composition of Formulation 15 including all of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and extract of Sesamum indicum in addition to the extract of Pinus koraiensis showed considerably increased wrinkle improving effect, as compared to the composition of Comparative Formulation 1 including only the extract of Pinus koraiensis.

[Experimental Example 5] Effect promoting the differentiation of keratinocytes

Human keratinocytes were cultured in a culture flask, grown to about 80% and treated with samples of Table 9 at a concentration of 5 ppm for 4 days. Then, a mixture of a denaturing agent such as urea or SDS (sodium dodecyl sulfate) and a reducing agent such as β -mercaptoethanol was added to the culture to remove proteins. The remaining cornified layer was measured for absorption at 310 run to analyze peptide concentration. The result obtained by comparing the samples to the negative control of distilled water (0%) is shown in Table 9. [Table 9]

From the result of Table 9, it was confirmed that the compositions of Examples 1 to 15 according to the present invention had excellent effect promoting differentiation of keratinocytes.

[Formulations 16 to 30 and Comparative Formulation 2] Preparation of cosmetic composition for skin moisturizing

The cosmetic compositions of Formulations 16 to 30 and Comparative

Formulation 2 were prepared using the extracts of Examples 1 to 15 and Comparative Example 1 according to the composition described in Table 10.

The aqueous phase of the Ingredient Nos. 1 to 5 and the alcoholic phase of the

Ingredient Nos. 6 to 9 in the composition of Table 10 were separately dissolved at room temperature. The alcoholic phase was slowly added to the aqueous phase, while stirring, for solubilization. Then, after the completion of the addition of the alcoholic phase, the reaction was further stirred for 3 to

5 minutes and deaerated in vacuum to complete the preparation of the cosmetic composition.

[Table 10]

[Experimental Example 6]

Effect recovering the barrier function of skin

In order to measure the effect recovering the barrier function of skin of Formulations 16 to 30 and Comparative Formulation 2, acetone was repeatedly applied on the skin of hairless mouse to damage the barrier function. When the transdermal water loss (TEWL) reached 4.0g/m²/h upon measurement by evaporimeter EPl (Servomed, Sweden), the samples of Formulations 16 to 30 and Comparative Formulation 2 were applied on 5 cm² area and the transdermal water loss was measured at 1, 2, 4 and 8 hours later to determine the reduced amount, thereby evaluating the recovering of the barrier function. Here, a known lipid mixture (mixture of ceramide, cholesterol and fatty acid (2:1 :1) was used as a positive control. The result was determined by change in the transdermal water loss according to time, based upon the transdermal water loss at the damaged barrier status (100%) and is shown in Table 11 below. [Table 11 ] Transdermal water loss (%)

From the result of Table 11 , it was noted that the compositions of Formulations 16 to 30 including at least one selected from the group consisting of the extract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum in addition to the extract of Pinus koraiensis were more effective than the composition of Comparative Formulation 2 including only the extract of Pinus koraiensis and when the kinds of the extracts selected from the group consisting of the extract of Prunus mume, the extract of Phaseolus angularis, the extract of Chaenomeles sinensis and the extract of Sesamum indicum added to the extract of Pinus koraiensis was increased, the effect recovering the barrier function was much greater. Now, Formulations of the cosmetic compositions according to the present invention are described. However, they are only for illustrative purpose and the present invention is not limited thereto.

<Soap formulation > [Table 12]

<Lotion formulation> [Table 13]

<Cream formulation> [Table 14]

<Pack formulation> [Table 15]

<Beauty liquid formulation> [Table 16]

[Mode for Invention] [Industrial Applicability]

As described above, the cosmetic composition according to the present invention comprises extract of Pinus koraiensis and at least one selected from the group consisting of extract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum to show the effect promoting the growth of keratinocyte, the effect removing oxygen free radicals produced by UV rays, the effect promoting biosynthesis of Type 1 procollagen by reducing the biosynthesis of MMP-I caused by UV rays associated with decomposition of skin substrates, the effect preventing aging and the effect moisturizing skin. Therefore, the composition may be used as a composition for promoting the growth of keratinocytes, a composition for removing oxygen free radicals, a composition for promoting biosynthesis of Type 1 procollagen by reducing the biosynthesis of MMP-I, a composition for prevention skin aging and a composition for moisturizing skin.

Claims

[CLAIMS] [Claim 1 ]

A cosmetic composition for promoting growth of keratinocyte comprising extract of Pinus koraiensis and at least one selected from the group consisting extract of Primus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient.

[Claim 2]

A cosmetic composition for removing oxygen free radicals comprising extract of Pinus koraiensis and at least one selected from the group consisting extract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient.

[Claim 3] A cosmetic composition for promoting biosynthesis of Type 1 procollagen by reduction of biosynthesis of MMP-I comprising extract of Pinus koraiensis and at least one selected from the group consisting extract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient.

[Claim 4]

A cosmetic composition for preventing aging of skin comprising extract of Pinus koraiensis and at least one selected from the group consisting extract of Prunus mume, extract of Phaseolus angularis, extract of

Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient.

[Claim 5]

A cosmetic composition for moisturizing skin comprising extract of Pinus koraiensis and at least one selected from the group consisting of extract of Prunus mume, extract of Phaseolus angularis, extract of Chaenomeles sinensis and extract of Sesamum indicum, as an active ingredient.

[Claim 6]

The composition according to any one of claims 1 to 5, in which the extracts are extracted with water as an extract solvent.

[Claim 7] The composition according to claim 6, which further comprises 1,3- butylene glycol. [Claim 8]

The composition according to any one of claims 1 to 5, in which the extracts are included in an amount of 0.0001 to 25 wt% based on the total weight of the composition.