TW200932756A - External skin treatment composition comprising silybin glycosides - Google Patents

Abstract

The present invention relates to a complex silybin glycosides external skin treatment composition; and further, it is an external skin treatment composition including silybin glycosides.

Description

200932756 VI. Description of the Invention: [Technical Field to Be Invented] The present invention relates to a composition for external use on skin. [Prior Art] It is known that difloxacin containing water as a main component is effective in preventing skin aging, in the treatment of erythema, burns, dystrophic state of skin or mucous membranes, dermatitis

Isochronization can effectively promote healing and can effectively protect the skin from external environment (radiation, wind, sun, etc.) (refer to the patent document 曰: 曰本特开平M〇〇132号), in addition, it is known that water can be used. The effect of inhibiting the differentiation of the epidermal keratinocytes of Libin (refer to Japanese Patent Laid-Open No. Hei 2_-_7), and the promotion of the production of type I collagen containing S. sinensis as a main component (see Patent Document 3: WO2004/85429). In addition, it is a technique for dispersing water lysine (see Patent Document 4: JP-A-Japan), because it is hardly soluble in water and oil. "Innocence Bulletin", Gu Shui Shuifei 15 Sustained _ _ complex, there is a technology to view its biological ship (3) = Li Literature 1: Japan made Ping Bu 1G_ bulletin), the formation of microemulsion composition = biological In the case of the singularity of the singularity of the singularity of the singularity of the singularity of the singularity of the product. P makes use of some techniques*, and sometimes does not completely inhibit the precipitation of the water fly Libin 200932756, and even if the precipitation of water _t can be suppressed, there is a restriction on the prescription cake. It is known that the sugar glycoside, galactose glycoside, lactose glycoside, and maltose glycoside have antioxidative effects, and are excellent in water and water. (Ref. 1 * Kosina P. et al. , Phytother. Res. 16, S33-S39 (2002)). However, it is not clear that the skin external composition containing the watery lycopene glycoside, the wrinkle formation inhibitory effect of the watery lycopene glycoside, the epidermal keratin cell differentiation inhibiting effect, and the type I collagen production promoting effect. [Patent Document 1] Japanese Laid-Open Patent Publication No. 2004-91397 (Patent Document 3) WO2004/85429 (Patent Document 4) Japanese Patent Laid-Open No. 2002-34505 [Patent Document 5] 曰本本表-2003-503441 [Patent Document 6] JP-A-3-206090 [Non-Patent Document 1] Kosina P. et al., Phytother. Res. 16, S33 - S39 ❹ (2002) [Explanation] An object of the present invention is to develop a skin external composition containing a silibinose glycoside as an active ingredient. The present inventors completed the present invention by using a silibinin glycoside. 4 200932756 That is, the main constitution of the present invention is as follows: (1) A skin external composition containing a silybin glycoside. (2) The composition for external use on skin according to (1), wherein the silybin glycoside is a silibinin glycoside of the formula (1) or a silibinin of the formula (2) The buds are glycosides.

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Formula (2) (3) An inhibitor of wrinkle formation in which a silybin glycoside is used as an active ingredient. 5 200932756 (4) An epidermal keratinocyte differentiation inhibitor in which a silibinin glycoside is used as an active ingredient. (5) A type I collagen production promoter, wherein a silibinin glycoside is used as an active ingredient. _ (6) A skin roughness improving agent for sun exposure, in which a silybin glycoside is used as an active ingredient. (7) The preparation according to any one of (3) to (6), wherein the silybin glycoside is a silybin milk glycoside of the formula (1) or a formula (2) This milk thistle® bin malt glyceose.

Formula (1) 6 200932756

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Formula (2) ❹

(8) The composition for external use on skin according to (2), or the preparation according to (7), which comprises water as a solvent and contains a silibinose glycoside 〇〇〇.〇〇〇8~5 〇 weight %. The effects of the present invention are as follows: 1. By providing a water-soluble 15 guest glycoside having high arsonability, it is possible to provide an exogenous skin composition, a wrinkle formation inhibitor, and an epidermal keratinocyte which enhance the mechanism of action of the water lycopene. The composition for external use, the composition for external use for preventing aging, the collagen-producing agent for sputum type, and the skin-improving agent for sun exposure. _ is an increase in water solubility, improved safety, low irritation, and expanded use. 2. In particular, it is effective as a silybin glycoside of the silybin formula, or a silybin maltoside glycoside represented by the chemical formula (2). 3. The silybin glycoside used in the present invention has high water solubility and can be used as a water-soluble type. It can be used as a skin external composition such as lotion, lotion, cream or mask. '10(s) silk milk, makeup m-training or cream-like 2 base, etc. 200932756 Cosmetic skin external composition, hand cream, leg warmer A body external composition such as a body lotion, a bathing agent, or the like. 4. The silybin glycoside used in the present invention can be dissolved in water. 〇〇〇8~5 0% by weight. Even in the case of an emulsion or the like, a large amount of the mixture can be blended in the aqueous portion. [Embodiment]

Silybin (CAS No. 22888-70-6) is from the genus Sylvestris sylvestris (scientific name Silibum marianum Gaertn, alias big cockroach, big-winged pheasant, chyle; CAS ❹ No. 84604-20-6 a kind of flavonol lignan extracted from the milk thistle, the flavonoid lignan extracted from the milk thistle is called silymarin (Casi. Cas No. 65666-07-1), in addition to water, and also contains water. Silydianin (CAS Να 29782-68-1), Sily lychristin (CAS No. 33889-69-9), Isosilybin (CAS No. 7258, 71-6) Wait. Shuifeiweibin can be separated from silymarin by chromatography, or it can be obtained by purchasing reagents. Ο Silibinin glycosides can be used according to the literature (KrenV. et al., j.Chem.Soc.,

Perkin Trans 1, 2467-2474 (1997)), using Lewis acid as a catalyst, is prepared by combining the sugar of the base with a water-based fish carp to protect the base sugar. In the reaction system, the sugar selectively forms a glycosyl bond with the hydroxyl group of the primary alcohol of the water. Using Lewis acid as a catalyst, by reacting silibinin and whole-bestinized lactose 8 200932756 A glycosidic bond is formed, which is deacetylated to obtain the silibinin milk glycoside of Wu (1).

Using Lewis acid as a catalyst, a glycosidic bond is formed by reacting silibinin and total acetylated maltose to deacetylate to obtain a silibinin maltose glycoside of formula (2).

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Formula (2) ❹ The present invention enables Guanshui _(4)(4) to be excellent in water solubility, and the water repellency and the water-spraying malt glycoside are soluble in water. Therefore, in the case of the lotion that cannot be used in the past, if it is a water shop, it can be used in conjunction with 0 9 200932756 in the scope of the effect of the county tree (4). Containing oil, surfactant, preservative, multi-tree, ethanol, sugar, metal ion sealing, water polymer, _, powder component, UV absorber, UV blocker, moisturizer, Spices, pH adjusters, etc. In addition, it may also contain other medicinal ingredients and physiologically active ingredients such as vitamins, skin activating agents, blood flow promoters, resident bacteria control agents, active oxygen scavengers, anti-inflammatory agents, whitening agents, and bactericides. The present invention makes (10) Shuifei 15 Bin@@ _ water-soluble excellent, so in the present invention, Shuifei Libin milk glycoside, Shuifei 15-bin malt glycoside can be dissolved in water. Therefore, in the past, it was difficult to match the water (4) lotion, and it was also possible to match the syrup. As the water-spraying agent of the present invention, the 15-glycoside-containing saccharide is an active ingredient-pattern formation inhibitor, and examples thereof include a composition for external use of water, Wei, cream, _, etc. Products, sensation improvement medicines, etc. Even in the case of emulsions, it is possible to mix the silybin glycoside at a high concentration in the aqueous portion. The differentiation of the epidermal dermal keratinocytes of the epidermal dermal keratinocytes, which is the active ingredient of the water-object (4) of the present invention, is proliferated, and the lion is prevented from ameliorating the delay of metabolic renewal, preventing aging, ultraviolet irradiation Since the skin of the epidermis is flattened and the aging skin acts again, it can be used as a skin external composition for preventing aging. I ^ Collagen with the water content of the sucrose as the active ingredient of the fx, the 200932756 promoter is used to improve the tension and impurities of the skin, and the lion can be expected to prevent and improve the leveling and relaxation. Anti-lang skin external composition is used. [Synthesis of Saccharomyces cerevisiae glycoside] The spirulina milk glycoside was synthesized according to the method of Helferich. _ In the presence of air, make water _ 宾 (3. Gg, 6. and Xin Yi waking base ^ lactose (6 · 3g, 9. 2mol) in 1_ of the two gas a hydox - acetonitrile (1: Bu v / v) In the granules, 14 ml of boron trifluoride dimethyl scale complex, and 1 M at room temperature ® yna. After the reaction, the solution was cooled with ice and a saturated aqueous solution of sodium carbonate was added. The extracting treatment was carried out twice with 150 ml of a two-gas ablation process. After the treatment with anhydrous sodium sulfate, the extraction solvent was removed by an evaporator. The triethylamine-nonanol-water (1 ··δ: υ was reacted at 35 〇 for 3 ,, Then, the solvent was removed by an evaporator. Purification was carried out using B0NDESIL_cl8 (Varian) to obtain a water-squeezed 4-negative lactoside (1. 〇g, yield 2% by weight). The MS spectrum of the obtained silybin milk glycoside was obtained. Confirmation of 'detection of mass spectrum peak of [M+H]+ : 8〇7 5 . ® [Synthesis of silibinin maltose glycoside] Synthesis of silibinin maltose glycoside according to Helferich method. In the presence of 'silibinin (3.0 g, 6.2 inol) and octyl-indole-D-maltose (β. 3g, 9.2 mol) in 180 ml of dichloromethane-acetonitrile (1: 1, v / v) solvent, and trifluoride The dimethyl ether complex (1.14 ml, 12.4 mmol) was stirred at room temperature for 19 hours. After the reaction was completed, ice-cooled, a saturated aqueous solution of sodium hydrogencarbonate was added, and extracted twice with 150 ml of dioxane. After the anhydrous sulfur 200932756 sodium sulphate treatment (four) hair removal remove the extraction solvent. The ethamine-methanol-water (1:8: 〇 at 35. 匸 reaction continent hours, then use the solvent to remove. 帛 _ESIL-C18 (Varian Purification was carried out to obtain a gas 麦 ( ( 四 四 Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Q Mass spectrometry peak of 5. [Example 1] Water bathing test 1 · Experimental method Weigh the appropriate amount of water philips, water (four) broth glycoside, and water saponin malt glycoside in 1. 5 ml of the tester (the following will Water Rybin is abbreviated as SB, water, fine milk, sugar, body, SBL, Spirulina, malt, glycoside, abbreviated as Μ), and added pure. Water purification 'matched into various concentrations' by visual inspection It is judged whether the appearance transparency and the centrifugation at room temperature 〇5_rpm, 5min), and whether it is water-soluble in SB. Poor, weighed small amount of water was added drop grant is stirred for about 1 hour ❹ grant 'to stop the mixing operation! Sink is found, it is determined that the house after hours was dissolved in a beaker. TABLE 1

----Compound name/concentration (mM) .__ 0.01 0.1 1 —----- 50 60 70 100 SB ---- △ XXXXXX SBL 〇〇〇〇△ XX SBM 〇〇〇〇△ X ' X 〇 : No precipitation X: Precipitation Δ: The result of the experiment of 200932756 was not determined. It was found that the water solubility of both SBL and SBM was sharply improved and the water solubility was improved to about 5,000 times or more compared with SB. The solubility of both SBL and SBM could not be detected. The SB concentration is 0. OlmM is equivalent to 0.005% (w/v). The concentrations of SBL and SBM 50 mM, 60 mM were equivalent to 4. 1% (w/v) and 4.9% (w/v), respectively. Therefore, if the solubility of SB, SBL, and SBM is compared by mass/capacity %, the solubility of SBL and SBM is increased by about 10,000 times compared with SB. In a preparation such as a water-soluble skin external composition, SB having a concentration of 〇. 0005% (w/v) has been difficult in the past, and β can now contain a SBL of 4.1% (w/v) concentration, which may contain 5 . 0% (w/v) concentration of SBM. Further, the concentration of SBL and SBM is 0. OlmM is equivalent to 000. 0008% (w/v; > [Example 2] Epidermal keratinocyte differentiation inhibition test, proliferation maintenance effect 1. Experimental material 1.1 Human normal epidermal keratinocyte ❷ The human normal epidermal keratinocyte NHEK (Asahi Techno Glass) was cultured in an epidermal keratinocyte culture medium: KGM (Asahi Techno Glass) in a 37 ° C - 5 % C 2 incubator. The experiment used a sub-algebra of 3 to 5 Generation of cells. 1.2 KGM (the medium for epidermal keratinocytes) KGM is a human epithelial cell proliferation factor (0. lng/ml), insulin (5. Ο/zg/ml), hydrogenated on the basal medium of epidermal keratinocytes. The medium after pine (〇. 5/zg/nii), Qingda Zhengsu (50//g/ml), amphotericin B (50/ig/ml), and bovine pituitary gland extracting 200932756 liquid (2 ml). When a sample represented by a silibinin glycoside is added to a cell, the experiment is carried out using KGM medium in which only the bovine brain pituitary extract is removed. 1.3 Adding a sample to Silibinin (SB) and water fly蓟宾麦芽配糠 (SBM), 水飞莉宾乳糖 (SBL) dissolved in DMS0 (two In the sulfoxide: Wako Pure Chemicals, it was added at various concentrations. 2. Experimental method 2.1 Epidermal keratinocyte differentiation inhibition test NHEK was suspended in KGM to make 5×10 4 /ml, and inoculated into 6-well culture plates at 4 ml/well. The cells were incubated for 24 hours, and the cells were adhered to the culture plate. KGM supplemented with each compound was removed at 4 ml/well, and the medium was changed every 2 days for 8 to 10 days. Morphology, when the surface-treated control cells showed morphological changes (flattening) of signs of differentiation, photographing was performed, and the culture was terminated. 2. 2 Epidermal keratinocyte proliferation maintenance test The cells obtained from the above test tissues were passed through the egg-like enzyme. After the plate was peeled off, it was suspended in KGM to be 2. 5_V〇a. The cell suspension was seeded at 2 ml/well on a 24-well culture plate. The medium was changed every 2 days and cultured for 8 days. After the culture, The 瞧 was peeled off from the culture plate by the egg (4) treatment, and the number of cells was determined by a Beckman-Coulter. 3. Experimental results 200932756 3.1 Experimental results obtained by the epidermal keratinocyte differentiation inhibition test are shown in Fig. 1, It is a micrograph of the cells. In comparison with the control group Control and SB 3 (cultured in the medium supplemented with silybin 3#M) of DMS0 treatment, the epidermal keratinocytes were flattened, and the morphological changes of signs of differentiation were revealed. On the other hand, there were no signs of differentiation in SBM3eM and SBL3//M. When cultured in a medium of SB, SBL, or SBM added separately, the differentiation of epidermal keratinocytes was all inhibited. SB, SBL, and SBM all have a differentiation inhibitory effect on epidermal keratinocytes, but SBM and SBL have superior epidermal keratinocyte differentiation inhibitory effects compared with SB. 3. 2 Epidermal keratinocyte proliferation maintenance test In the above-described epidermal keratinocyte differentiation inhibition test, if differentiation is inhibited, the cells should maintain proliferative ability and can be sequentially propagated by subculture. Cells that are induced to differentiate cannot proliferate because they are irreversible reactions. Then, the cells obtained in the above test were subcultured, and the proliferative ability maintained was examined by measuring the number of proliferating ® cells. When the sample is at an hour, the cells after adding SB are added.

The number of cells increased to 2.1 times, and the results of SBL and no addition are shown in Fig. 2. The sample thickness j thinning ability is hardly maintained, Sbm and; to 1.8 times, SBL is compared with no sample added, i Compared with the sample of 200932756, the number of cells increased to L8 times, and it was confirmed that the cell proliferation ability was maintained. Compared with SB, it can be seen that the maintenance effect of cell proliferation ability of SBM and SBL is high. [Example 3] Type I collagen production promoting effect 1. Experimental material L1 Human skin fibroblast cells Human skin fibroblasts CCD1074SK (Daichi Sumitomo Pharmaceutical Co., Ltd.) were cultured in d_Mem with 37 C-5% C〇2 incubator . In this experiment, cells with sub-algebras of 丨〇-15 generations were used.

1.2 D-MEM D-MEM, 10% fetal serum (Hyclone) was added to D-MEM basal medium (GIBC0) and used. Further, when the sample was processed, the experiment was carried out using D-MEM without adding bovine fetal serum. 2. Experimental method Human skin fibroblast CCD1074SK was suspended in D-MEM medium containing 1% fetal calf serum, which was made into 3xlGVml, inoculated in a culture dish, and cultured for 24 tons. On the board. The squama was replaced with (iv) μ medium in which each of the minerals in DMS0 was added to each of the concentrations of Kodak fetal serum. The medium was incubated in the medium for 48 hours, and the culture medium was concentrated. After concentrating 5 〇 _ or less, the egg was quantified, and the egg 200932756 white matter was collected, and the sample was concentrated as a cell culture solution for western blotting. Using a protein of 1 泳gg per lane, using SDS_pAGE After separation, it was transferred onto a nitrocellulose membrane. The transferred nitrocellulose membrane was immersed in a blocking solution (in a pBS containing %. 1% polyoxyethylene (20) sorbitan monolaurate dissolved in skim milk powder to a 5% concentration solution) °c closed - staying up late. Wash with sodium sulphate (containing (% polyoxyethylene (20) 6 linoleic acid monolaurate (tetra) pBS); after the strip, immerse in a _ under antibody [prepared into 5 ng / ml with stripping solution In the polyclonal antibody of type I collagen (Rockland), at room temperature, after silking, impregnated with secondary antibody (prepared with 25 ng/ml of horseradish peroxidase-labeled anti-immunization with washing solution) In the globulin G), the reaction was carried out for 1 hour at room temperature, and after washing with a Western blotting detection reagent (Amersham Biosciences), the results of the test were verified to be the same as the SB. Both SBM have the promotion of i-type collagen production. The experimental results are shown in Fig. 3. The intensity of the obtained band is passed through the program software Image; the results of digitization are shown in Table 2, and the D of DMS0 treatment The amount of collagen produced as a comparative yield when 丨 is shown in Figure 4. Table 2 200932756

DMSQ SB3|jiVl SB1〇mJV1 SBM3_ SBMIC^JVI SBL3mJV1 SBLICVJVl ^5.064 831656 8397 9S977 116102 &4.068 107.797 rBlative irtensitv 1.00 1.86 1.86 ZC9 258 2C9 239 indicates that SB, ·, and SBL all have type I collagen production promoting effects. Compared with the ministry, the ignorance of the fox and the fox is selfish. When the sample concentration is 1 MM, the effect of SBM and SBL is large, and it is remarkable when a high concentration is confirmed. [Example 4] Inhibition of water loss by ultraviolet irradiation, inhibition of formation of wrinkles L experimental materials, apparatus, experimental animals, hairless mice, H〇s; HR1, 5 weeks old (Hoshino experimental material) L2 ultraviolet irradiation device Ultraviolet A wave (FL32SBL/DMR: manufactured by Clinical Supply Co., Ltd.) 200932756 Ultraviolet B wave (FL32SE/DMR: manufactured by c 1 ini ca 1 supp丨y) 1. 3 Percutaneous water loss measuring device

Vapometer (manufactured by k-science) 1.4 Replica collection instrument and replica analysis system

(Yes) Reflective replica collection kit and replica analysis system manufactured by Asahibiomed Co., Ltd. ASA-03RXD 2. Experimental method 2.1 2.1 Feeding environment at 25 °C ± 2 °C, humidity 50% ± 5%, freely ingestible It is raised in a conventional breeding environment as a food feed MR and tap water. Each component was 5 ‘in the same cage. • The photo is from 7 am to 7 pm and is set to stay up every 12 hours. 2.2 UV irradiation The hairless small milk Hos; HRl, after training for 1 week, began to irradiate ultraviolet rays. Hairless mice were transferred to a special cage under UV-ray irradiation, and each group was irradiated with UVB of UVB20 mJ/cm2 and UVAlOJ/cm2. Irradiation was performed on Monday, Wednesday, and Friday, and was cycled 3 days a week for 1 week, and 30 times of ultraviolet rays were irradiated. 2. Three sets of settings After UV irradiation, the surface of the back skin of the mouse was treated with 100//L SB, SBL, SBM sterol solution or solvent (sterol) within 30 minutes. The concentrations of SB, SBL, and SBM coating were set to 丨〇%, 〇3%, 〇1%, respectively. 3 200932756 For all groups, UV (four) for mice coated with solvent methanol, UV-irradiated group and UV Irradiation group. The use of methanol as a solvent is to dissolve the SB in the sterol, and the fox is completely dissolved at the concentration of (9), but π is only in the Z shoufan, the glutinous solution, and the 解. 3% when there is some precipitation, which is found at 1%. I can't do anything. Even when insoluble matter was found in the SB methanol solution, it was directly used for the experiment. 2. 4 Determination of transepidermal water loss The measurement of transepidermal water loss is performed using Vap〇Meter (Keys1; 〇ne entific system)' for the tail from the back to the head direction, from the lumbosacral vertebrae The portion of the right G·5αη was measured three times, and the average value was determined. The Nail model was used to measure the opening at the end of the measurement (the opening was narrowed to correspond to the narrow range of the mouse skin), and it took about 19 seconds for each i-human measurement time. The measurement day was carried out after 1 week of ultraviolet irradiation. 2. 5 Replica image analysis In order to correctly grasp the formation of wrinkles, collect replicas. The replica image analysis was carried out using a reflection replica analysis system, ASA-Q3RXD (manufactured by Asahibiomed). Use the ASA-03RXD 'to illuminate the captured replica from a 27 degree angle to illuminate parallel light (LED light source)'. The shadow image corresponding to the obtained wrinkle shape is imaged by a CCD camera, input to a computer and graphed Like the treatment, the wrinkle volume ratio (#m3/mm2/100) on the surface of the replica was measured. 2. 6 Statistical analysis The test results 帛 mean ± fresh deviation (S. D ·) indicates that the significant difference test 疋 is tested by the Bartlett test-^x method for homoscedasticity test. No rejection of homosexuality leave 20 200932756 At the time of the Dunneti: multiple test, the Dunnett multiple test was used as a reference material when rejecting the same variance hypothesis. 3. Test results 3.1 Changes in body weight and appearance After the end of the irradiation for 10 weeks, there was no significant difference in body weight change between the groups. There were also no mice showing severe lesions. As a result of observing the appearance of the mouse skin, it was found that the ultraviolet irradiation® sterol treatment group of group 2 had wrinkle formation, while the group 4 of 0.3% silibin coating group, group 6 to group 8 of all SBM coating. In all of the SBL-coated groups of the group, the group 9 to the group 11, wrinkle suppression was observed. 3. 2 Percutaneous water loss As an indicator of skin roughness, the amount of water loss in each group was measured using VapoMeter after 1 week of ultraviolet irradiation. The results are shown in Table 3 and Figure 5. Compared with the group 1 ultraviolet non-irradiated group, the percutaneous water fraction of the ultraviolet irradiation group of group 2 lost the increase of f. In group 3 to group 11 of the water silibin (SB) and the silybin sucrose (SBM'SBL) coating group, the significant level of 1% or less significantly inhibited the increase in transepidermal water loss. When the water loss amount of one set (preparation, external line non-irradiation, and decyl alcohol coating) is 〇%, and the water loss amount of two sets (preparation, external irradiation, and decyl alcohol coating) is 1 (10)%, 3 to 5 groups (prepared) The amount of water loss in the outer line irradiation, the part 〇. 1~1% coating) is Shen~π%, and the water content of the 6~8 group (ultraviolet irradiation, SBM 〇1~1% coating) is 200932756 2W '9~丨丨The moisture content of the group (ultraviolet radiation, fox Q. coating) was 24 to 27%. By adding SB, SBM, and SBL, the increase in water loss was suppressed. In addition, it is the effect of SBM and SBL of water-based saccharide body to suppress the increase in water loss. In short, it has the effect of improving the roughness of the skin caused by the sun. Table 3 " Group No. UV Coating Concentration (w/v) Moisture Evaporation (g/m2h) —-— Moisture Evaporation Ratio (%) 1 Non-irradiation Methanol —— 11.8 0 — 2 Irradiation of Methanol - ~~ — 25.0 100 3 Irradiation SB methanol solution 0.1 ---- 19. 2 56 4 m hx. SB sterol solution 0.3 ----- 阳, shot 15.2 2B 5 Irradiation SB sterol solution l7 〇~ ---- 21.8 76 6 Irradiation of SBM methanol solution ------- 0.1 14. 4 20 7 Irradiation of SBM methanol solution 0.3 ---- 14. 6 21 8 Irradiation of SBM methanol solution '~~~---- - 1.0 13.2 11 9 Irradiation of SBL methanol solution 0.1 15.0 24 10 Irradiation of SBL methanol solution 0.3 -------- 15.3 27 11 Irradiation of SBL methanol solution 1.0 ------ 15.0 24 ---

3·3 Wrinkle volume ratio 1 After the end of the irradiation period, 'To collect the replicas correctly, collect the replicas' and measure the wrinkle volume ratio of the surface of the replica by image analysis ("heart_2/1〇〇). The results are shown in Table 4 and Figure 6. The results of Dunnett's significant difference test were carried out, bribe in group 2 (UV 22 200932756 irradiated f alcohol coating), group 1 (ultraviolet non-irradiated methanol coating), group 4 (ultraviolet SB 〇 〇 coating), group 7 (ultraviolet The illuminating volume was significantly suppressed at a significant level of 5% or less, respectively, in Group ii (UV-irradiated SBL 1. 〇% coating). Table 4 Group No.

UV coating concentration % (w/v) Ο Ο 10 —— 11 Non-irradiation irradiation - One irradiation - Irradiation wrinkle volume ratio (;zm3/mmVl〇〇) Methanol sterol SB methanol solution SB sterol solution SB methanol Solution SBM methanol solution SBM sterol solution SBM methanol solution SBL methanol solution SBL methanol solution SBL methanol solution 12 17 20 19 14 [Prescription example 1 lotion] ^% by mass • Water _Bin milk glycoside 0 3:. ) Diglycerol consists of 5.0 •1,3~butanediol 23 200932756 4. (monocondensed) dipropylene glycol 3.0 5. Potassium hydroxide amount 6. citric acid amount 7. Purified water remaining _ (preparation method) Dissolve 1 in 7 6. [Prescription Example 2 Emulsion] Mass % 1. Silibinin malt glycoside 0.3 2. Hydrogenated soybean malaise 0.7 3. Decaglyceryl stearate (HLB 12) 2.0 4. Glycerin 8.0 5. Olive oil 8.0 6. Beryl alcohol 1.0 7. (-) dipropylene glycol 8.0 8. Carbopol 0.1 9. Xanthan gum 0.2 10. Potassium hydroxide amount 11. Molybdic acid amount 12. Pure water remaining

❹ 24 200932756 (Preparation method) 1 to 4 and 7 to 12 are dissolved at 80 ° C. Adding to it has been heated to about ❹

5 and 6 at 80 ° C, stirred and mixed with a homogenizer, and cooled to 30 ° C to obtain an emulsion. [Prescription Example 3 Moisturizing Beauty Liquid]% by mass 1. Silibinin milk with sugar body 0.2 2. Hydrogenated soybean fat 0.6. Decaglycerol monooleic acid S (HLB 12) 1.5 4. Glycerin 7.0 5.1, 3 - Butanediol 5.0 6. Polyethylene glycol 4000 0.1 7. Squalane 5.0 8. Chopped 0.5 9. Two (phytosterol / octyldodecanol) Laurel glutamate 0.2 10. Xanthan gum 0.3 11. Potassium oxyhydroxide amount 12. Capsitric acid amount 13. Purified water remaining (preparation method) Dissolve 1 and 11~13, add 4~6, 10, and then heat to 80 °C to dissolve. 25 200932756 2, 3, 7 to 9 which had been heated to about 80 ° C were added thereto, and cooled to 30 ° C to obtain a moisturizing cosmetic liquid. [Prescription Example 4 Moisturizer] —% by mass 0.5 10.0 〇 8.0 0.5 0.01 0.5 10.0

1. Silibinin malt glycosides 2. (-) diglycerin 3 · (-) dipropylene glycol 4.1, 2-pentanediol 5. L-serine 6. Decaglycerol distearate (HLB9. 5) 7. Decaglycerol monomyristate (HLB14) 8. Eucalyptus oil 9. Macadamia oil 10. Mountain alcohol 11. Oxanone 12. Jojoba oil

13. Vitamin E 14· SIMULGEL NS (manufactured by SEPPIC) 15. Xanthan gum 26 200932756

16. Potassium hydroxide Appropriate amount 17. Citric acid Appropriate amount 18. Purified water Remaining (preparation method) Dissolve 1 and 16~18, add 2~5, and then heat to about 80 °C to dissolve. Add 6 to 14 which has been warmed to about 80 ° C, and cool to 30 ° C to obtain a moisturizer £ [Prescription Example 5 Body lotion] Mass % 1. Silibinin milk glycoside 0.2 2. PEG-60 hydrogenated castor oil (HLB14) 1.5 3. Glycerin 9.0 4. (monocondensed) dipropylene glycol 7.0 5. Sodium hyaluronate 0.001 6. Fluid stone soil 10.0 7. Anthrone 3.0 8. Octyldoxitol 4.0 9. (Acrylic/Acryl Acrylate (C10-30)) Ester Copolymer 0.2 10. Potassium Hydroxide Appropriate 11. Molybdenum Acid 12. Purified Water Remaining 27 200932756 13. Ethanol 2.5 (Manufacturing Method) 1 and 10~12 Stir and dissolve, add 2~5, 9 and then warm to about 80 ° C to dissolve. 6 to 8 which had been heated to about 80 ° C was added thereto, cooled to 30 ° C, and 13 was added to obtain a body lotion. [Prescription Example 6 Massage Cream]% by mass 1. Silibinin malt glycoside 0.05 2. Glycerin 10.0 3. (--) Diglycerin 2.0 4. Propylene glycol 7.0 5. Decaglycerol monostearate (HLB12 1.0 6. Cetyl ethylhexanoate 12.0 7. Samanol 2.0 8. Stearic acid 0.5 9. Sepinov EMT10 (manufactured by SEPPIC) 0.5 10. Flavor amount 11. Phenoxyethanol 0.3 12. Potassium hydroxide Appropriate amount 13. Moxibusty amount

28 200932756 14. Purified water Remaining (preparation method) Stir and dissolve 1 and 11~14, add 2~4, and then heat to 8CTC to dissolve. Mainly, 5 to 9 which had been heated to about 80 ° C was added, cooled to 30 ° C, and 10 was added to obtain a massage cream. 〇 [Prescription Example 7 emulsified foundation]% by mass 1. Silibinin milk glycoside 0.1 2. Glycerin 10. 0 3. (--) dipropylene glycol 8. 0 4.1, 2-pentanediol 1.0 5. Yellow Raw rubber 0.3 ® 6. Polyglycerol-2 triisostearate 1.0 7. Cyclome lime 8.0 8. Anthrone 5.0 9. Isostearyl neopentanoic acid vinegar 5. 0 10. Isostearic acid 1.5 11. Ammonic alcohol 0. 5 12. Dextrin palmitate 1.0 29 200932756 13. Talc 3.0 14. Titanium dioxide 5. 0 15. Iron oxide red 0.5 16. Iron oxide yellow 1.4 17. Iron oxide black 0.1 18. Potassium Hydroxide Appropriate amount 19. Molybdenum acid amount 20. Purified water remaining (preparation method) Stir and dissolve 1 and 18~20, add 2~5, and then heat to about 7 (rc dissolve. Then add fully pulverized 13 ~17 and mix and mix. Add 6~12 which has been warmed to about 80 °C, and cool to 30 °C to obtain an emulsified foundation. [Simplified illustration] Figure 1 shows the inhibition test of keratinocyte differentiation of epidermis Results Fig. 2 shows the results of the epidermal keratinocyte proliferation maintenance test. Fig. 3 shows the results of the type I collagen production promotion test. Fig. 4 shows the band shown in Fig. 3. Fig. 5 shows the numerical value of the amount of water loss in the mouse subjected to the ultraviolet irradiation test. Fig. 6 shows the surface of the skin replica of the mouse subjected to the ultraviolet irradiation test. 200932756 Volume ratio. [Main component symbol description]

Claims (1)

200932756 VII. Patent application scope: 1. A skin external composition containing silybin glycoside. 2. The composition for external use on skin according to claim 1, wherein the silybin glycoside is a silybin milk glycoside of the formula (1) or a water fly of the formula (2) Manbin malt with sugar body. OH 〇H OH Formula (1) OH Formula (2) 3. A wrinkle formation inhibitor in which a silybin glycoside is used as an active ingredient. An inhibitor of epidermal keratinocyte differentiation, wherein a silybin glycoside is used as an active ingredient. 32 200932756 5. A type I collagen production promoter in which a silybin glycoside is used as an active ingredient. 6. A skin roughness improving agent for sun exposure, wherein the silibinin glycoside is an effective ingredient. 7. The preparation according to any one of claims 3 to 6, wherein the water sulphate I1] negative glycoside is the silibinin glycoside of the formula (1) or the formula (2) ) Silibinin malt with glycoside. Formula (2) 8. The preparation described in the above-mentioned application, which is incorporated herein by reference in its entirety, which is incorporated herein by reference. 0% by weight of silybin with glycoside. 34