JP4648669B2 - Silymarin-containing cosmetics - Google Patents

Description

  The present invention relates to a cosmetic containing silymarin.

Silymarin is useful in preventing aging, promotes healing in the treatment of erythema, burns, dystrophic conditions of the skin or mucous membranes, dermatitis, etc., and protects the skin from external environmental stimuli (radiation, wind, sun, etc.) It is known that it is useful to do (patent document 1). In addition, silymarin suppresses sebum secretion (Patent Document 2), enhances epidermal permeation barrier (Patent Document 3), treats psoriasis and atopic dermatitis (Patent Document 4), improves epidermis flattening (Patent Document 5) )It has been known. Further, the present applicant has been continually researching silymarin, and has filed a patent application for the effect of promoting the production of type I collagen and elastin (Patent Document 6).
On the other hand, since silymarin hardly dissolves in water, it has a problem that it is difficult to mix it in an aqueous composition, and a technique of a beverage in which silymarin is dispersed is disclosed (Patent Document 7). Moreover, the technique which makes biocompatibility advantageous by making silymarin into a phospholipid complex is disclosed (patent document 1). In addition, a technique for improving the bioavailability by using a microemulsion composition has been disclosed (Patent Document 8). However, a silymarin solubilization technique for increasing the bioavailability of silymarin or preventing the precipitation of silymarin in transparent cosmetics is not known.

Japanese Patent Laid-Open No. 1-100132 JP 2000-169332 A JP 2000-169328 A JP-A-5-286864 JP 2004-91397 A International Patent Application PCT / JP2004 / 3978 JP 2002-34505 A Special table 2003-503441 Japanese Patent Publication No. 63-41396 Natural Drug Encyclopedia, edited by Takuo Okuda, Yodogawa Shoten, March 3, 1986 Wagner, H.C. , Et al. , Arznein. Forsch, 18, 696, 1968. Wagner, H.C. , Et al. , Arznein. Forsch, 24, 466, 1974. Titel, G.M. , Et al. , J .; Chromatogr. , 135, 499, 1977. Titel, G.M. , Et al. , J .; Chromatogr. 153, 227, 1978. Quercia, V.M. , Et al. , Chromatography in Biochemistry, Medicine and Environmental Research, Frigerio A. et al. (Ed). , Elsevier Scientific Publishing Company, Amsterdam, 1983, p1.

In order for silymarin blended in cosmetics to effectively act on the skin, it is essential that the silymarin is dissolved and penetrates into the skin. By simply dispersing silymarin as a fine solid in the cosmetic, silymarin does not penetrate the skin, and the effect of silymarin cannot be expected.
Silymarin dissolves in specific polyhydric alcohols, but when water is added, it immediately becomes cloudy and silymarin precipitates. It is not completely soluble in ethanol and some insolubles are produced. Moreover, when water is added, it becomes cloudy immediately when it is dissolved in the polyhydric alcohol. Silymarin can be solubilized if a high concentration surfactant is used, but it is not practical because it is too sticky. In addition, when the concentration of the surfactant is high, skin irritation occurs and there is a problem in safety. Reduction of surfactant is an important issue, and there is a demand for cosmetics with a minimal amount of surfactant. The present inventor has already applied for the effect of promoting production of silymarin type I collagen and elastin (Patent Document 6). ), And faced the problem that silymarin was difficult to dissolve in an aqueous base. Most of the prior art formulation examples do not solubilize and disperse silymarin. In such formulations, silymarin skin penetration cannot be expected, and the superior effects of silymarin cannot be obtained.
The object of the present invention is to provide cosmetics that solubilize silymarin, reduce the amount of surfactant used, and have excellent effectiveness and stability over time, as well as excellent usability and safety. It is to be.

The main configuration of the present invention is as follows.
1. A cosmetic comprising the following (1) to (5) and not containing ethanol .
(1) Silymarin (2) One or more polyhydric alcohols selected from polyethylene glycol, dipropylene glycol, 1,3-butylene glycol, 1,2-pentanediol, propylene glycol (3) For the amount of silymarin 0.5 to 6 times the amount of the surfactant , the surfactant is polyoxyethylene hydrogenated castor oil and / or polyoxyethylene polyoxypropylene alkyl ether,
(4) Strong base (5) Water
2 . 1. The concentration of silymarin is 0.7% by mass or more . Cosmetics described in 1.
3. The pH is 6.0 to 9.5. Or 2. Cosmetics described.
4). The pH is from 7.5 to 9.5. To 3. Cosmetics in any one of.
5. 4. above . A two-pack type cosmetic characterized in that the cosmetic described in 1 and an acidic cosmetic having a pH of 3.0 to 6.0 are separately filled in a container and mixed for use.

It was possible to provide a cosmetic in which silymarin was dissolved with a reduced amount of surfactant, less stickiness, and excellent usability and safety.
By incorporating a strong base, the solubility and dissolution stability of silymarin could be improved. By solubilizing silymarin, the function of silymarin is effectively exhibited by improving transdermal absorbability.
It becomes possible to solubilize 0.7% or more of silymarin, and to effectively express type I collagen production promoting action and elastin production promoting action.
By blending a specific polyhydric alcohol, a surfactant, a strong base, and water in combination, it was possible to solubilize silymarin more than 6 times the blending amount of the surfactant.
In order to adjust the pH to 7.5 to 9.5 to enhance the dissolution stability of silymarin, the cosmetic can be used as a two-part cosmetic used in combination with an acidic cosmetic having a pH of 3.0 to 6.0.

Silymarin is an invention in which it has been found that type I collagen production promoting action and elastin production promoting action are remarkable, and this action has been put to practical use in cosmetics. The concentration of silymarin blended in the cosmetic of the present invention is preferably 0.1% or more, and more preferably 0.7% or more. When the concentration of silymarin is 0.7% or more, type I collagen production promoting action and elastin production promoting action are remarkable, and when it is 0.7% or more, the permeability to the dermis is rapidly increased. The present invention is one in which the transdermal absorbability of this silymarin is enhanced and the usability, safety and stability as cosmetics are enhanced. The present invention solubilizes poorly soluble silymarin by blending a specific polyhydric alcohol, a surfactant, a strong base, and water, and reduces the blending amount of the surfactant used at that time. The present invention provides cosmetics that are excellent in stability and stability over time, and are excellent in usability and safety.
The cosmetic is preferably pH 6.0 to 9.5, more preferably pH 7.5 to 9.5.
It is also proposed that acidic cosmetics having a pH of 3.0 to 6.0 and other compositions are separately filled in a container to form a two-pack type cosmetic and mixed for use.

Silymarin (CAS No. 65666-07-1) is a flavono extracted from the asteraceae Maria Thistle (scientific name Silibam marianam Gaertn, also known as thistle, giant thistle, milk thistle; CAS No. 84604-20-6). It is a generic term of lignan, represented by the molecular formula _C 25 _H 22 _{O 10,} silybin (silybin; CAS No.22888-70-6), silidianin (silydianin; CAS No.29782-68-1), silichristin (silychristin CAS No. 33889-69-9), isosilybin (CAS No. 72581-71-6) and the like (Non-patent Document 1). In the present invention, the composition containing these flavonolignans contained in the extract of thistle is called silymarin as in the prior art. Silymarin is a mixture of flavonolignans as described above, and the content of plant extracts and plants as silymarin is determined by a method based on measurement with a spectrophotometer (Non-patent Document 2), a method by thin layer chromatography ( Non-patent document 3), and a method by high-performance liquid chromatography (non-patent documents 4 to 6). Among these measurement methods, 2,4-dinitrophenylhydrazine analysis, which is one of the methods based on spectrophotometer measurement, has been reported to the German Pharmacopoeia (monograph on the fruit of Sillybum marium) It is used. Also in the present invention, when determining the mixed composition of the above components, the content of silymarin was determined in terms of silybin using 2,4-dinitrophenylhydrazine analysis.

As a method for isolating silymarin with high purity from the fruit of thistle, methods of isolating with a purity of 70 to 80% and a method of isolating with a purity of 90 to 96% (Patent Document 9) have already been reported. . Silymarin is usually marketed as an extract raw material that is extracted from the seeds of Maria thistle with ethanol, ethyl acetate, acetone, etc., and obtained as a dry powder by spray drying. Silymarin used in the present invention is prepared in this manner, and commercially available silymarin can be used as it is. In addition, extracts obtained by concentrating constituents of silymarin such as silybin, silydinine, silycristin, and isosiribine from Maria thistle, and those isolated and purified can be used as compounds.
The plant body containing silymarin in the present invention can be used in all parts such as leaves, stems, buds, flowers, woody parts, bark parts (bark), ground parts such as roots, tubers, seeds, resins, etc. is there.
The silymarin and the plant body containing the same in the present invention can be used as a dried product obtained by drying itself and a dissolved product obtained by dissolving them using various solvents. For example, it can be used as a dissolved product using water or alcohols such as ethanol and methanol, polyhydric alcohols such as propylene glycol and 1,3-butylene glycol, and organic solvents such as ether, acetone and ethyl acetate.
The plant body containing silymarin in the present invention can be used as it is by natural drying, hot air drying, freeze drying, or fermentation. Moreover, when preparing a plant extract, what was obtained by performing processes, such as extraction, concentration, and pulverization, according to a conventional method can be used.
The concentration of silymarin blended in the cosmetic of the present invention is preferably 0.1% or more, and more preferably 0.7% or more. As shown in Test Examples 1 to 3, in a three-dimensional human skin model, a type I collagen production promoting effect and an elastin production promoting effect are remarkable at a silymarin concentration of 0.7% or more, and to the dermis at 0.7% or more. This suggests that the penetration of

The strong base used in the cosmetic of the present invention is a base having a high degree of ionization, and examples thereof include sodium hydroxide, potassium hydroxide, calcium hydroxide, and barium hydroxide. Of these, sodium hydroxide and potassium hydroxide are excellent in solubility and stability over time when silymarin is dissolved. By dissolving silymarin in an aqueous solution that has been made basic with a strong base in advance, it is possible to effectively solubilize silymarin that is hardly soluble in water.
As the silymarin dissolves, the pH of the system decreases, but it is necessary to add a strong base in advance so that the final pH of the cosmetic is 6.0 or higher. When the pH of the cosmetic is less than 6.0, silymarin is insufficiently dissolved, and silymarin is likely to precipitate over time. Moreover, when the pH of the cosmetic is adjusted to 7.5 or more, the stability over time is particularly excellent.
When the cosmetic is made basic, the dissolution stability of silymarin is improved, but the problem of rough skin occurs. As a means for applying basic cosmetic material to the skin with weak acidity, there is a two-component cosmetic that can be mixed with acidic cosmetic immediately before use. At that time, since cosmetics having a pH exceeding 9.5 have a too high pH, even if a two-component cosmetic is used, there is a problem in safety during production and use, and it is not suitable as a cosmetic.

Examples of the surfactant of the present invention include nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, naturally occurring surfactants, and the like. Among them, nonionic surfactants are preferable from the viewpoint of safety.
Nonionic surfactants include, for example, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene hydrogenated castor oil, polyglycerin fatty acid esters, sucrose fatty acid esters, polyoxyethylene polyoxypropylene alkyl And ethers. Of these, polyoxyethylene hydrogenated castor oil and polyoxyethylene polyoxypropylene alkyl ethers are particularly excellent in the solubility of silymarin.
Examples of the anionic surfactant include fatty acid salts such as sodium laurate, higher alkyl sulfates such as sodium lauryl sulfate, alkyl ether sulfates such as POE lauryl sulfate triethanolamine, N-acyl sarcosine acid and sulfosuccinic acid. Salts, N-acylamino acid salts and the like.
Examples of the cationic surfactant include alkyltrimethylammonium salts such as stearyltrimethylammonium chloride, benzalkonium chloride, and benzethonium chloride.
Examples of amphoteric surfactants include betaine surfactants such as alkyl betaines and amide betaines.
Examples of naturally occurring surfactants include lecithin, hydrogenated lecithin, lysolecithin and the like.
By adding a surfactant, the solubility of silymarin and the stability of the dissolved state are improved, but stickiness and skin irritation problems occur when the concentration of the surfactant is high. The concentration of the surfactant of the present invention can be 6 times or less the concentration of silymarin. The concentration of the surfactant is 0.5 times or more the concentration of silymarin. When the concentration of the surfactant is less than 0.5 times the concentration of silymarin, the effect of solubilizing silymarin is insufficient.

  The polyhydric alcohol used in the cosmetic of the present invention is polyethylene glycol, dipropylene glycol, 1,3-butylene glycol, 1,2-pentanediol, propylene glycol, and these polyhydric alcohols have an effect of dissolving silymarin. high. These can be used alone or in combination of two or more. Although other polyhydric alcohols can be blended, dissolution of silymarin is hardly promoted even when glycerin, diglycerin or the like is blended.

  In the cosmetics of the present invention, oils and fats such as vegetable oils, hydrocarbons such as waxes, higher fatty acids, higher alcohols, silicones, preservatives, saccharides, sequestering agents, water-soluble, as appropriate, depending on the application form A polymer such as a polymer, a thickener, a powder component, an ultraviolet absorber, an ultraviolet blocker, a moisturizer such as hyaluronic acid, a fragrance, a pH adjuster, and the like can be contained. In addition, vitamins, skin activators, blood circulation promoters, resident bacteria control agents, active oxygen scavengers, anti-inflammatory agents, whitening agents, bactericides, and other medicinal components and physiologically active components can also be contained.

  Examples of oils and fats include camellia oil, evening primrose oil, macadamia nut oil, olive oil, rapeseed oil, corn oil, sesame oil, jojoba oil, germ oil, wheat germ oil, glycerin trioctanoate, and the like, cocoa butter, coconut oil, Hardened coconut oil, palm oil, palm kernel oil, owl, owl kernel oil, hardened oil, hardened castor oil and other solid fats, beeswax, candelilla wax, cotton wax, nutca wax, lanolin, lanolin acetate, liquid lanolin, sugarcane wax, etc. Waxes.

Examples of the hydrocarbons include liquid paraffin, squalene, squalane, and microcrystalline wax.
Examples of higher fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and the like.

  Examples of the higher alcohol include linear alcohols such as lauryl alcohol, stearyl alcohol, cetyl alcohol, and cetostearyl alcohol, and branched chain alcohols such as monostearyl glycerin ether, lanolin alcohol, cholesterol, phytosterol, and octyldodecanol.

  Examples of silicone include linear polysiloxanes such as dimethylpolysiloxane and methylphenylpolysiloxane, and cyclic polysiloxanes such as decamethylcyclopentasiloxane.

Examples of preservatives include methyl paraben and ethyl paraben.
Examples of the sequestering agent include edetates such as disodium ethylenediaminetetraacetate, edetic acid, and sodium edetate.
Examples of polymers include gum arabic, gum tragacanth, galactan, guar gum, carrageenan, pectin, agar, quince seed, dextran, pullulan, carboxymethyl starch, collagen, casein, gelatin, methylcellulose, methylhydroxypropylcellulose, hydroxyethylcellulose, carboxymethylcellulose Examples thereof include vinyl polymers such as sodium (CMC), sodium alginate, and carboxyvinyl polymer (such as CARBOPOL).

  Examples of the thickener include carrageenan, tragacanth gum, quince seed, casein, dextrin, gelatin, CMC, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxyvinyl polymer, guar gum, xanthan gum, sodium carboxymethyldextran, bentonite and the like.

  Examples of the powder component include talc, kaolin, mica, silica, zeolite, polyethylene powder, polystyrene powder, cellulose powder, inorganic white pigment, inorganic red pigment, titanium oxide coated mica, titanium oxide coated talc, and colored titanium oxide coated mica. And organic pigments such as Red No. 201 and Red No. 202.

Examples of the ultraviolet absorber include paraaminobenzoic acid, phenyl salicylate, isopropyl paramethoxycinnamate, octyl paramethoxycinnamate, 2,4-dihydroxybenzophenone, and the like.
Examples of the ultraviolet blocking agent include titanium oxide, talc, carmine, bentonite, kaolin, and zinc oxide.

  Examples of the humectant include xylitol, maltitol, maltose, sorbitol, glucose, fructose, sodium chondroitin sulfate, sodium hyaluronate, sodium lactate, pyrrolidone carboxylic acid, and cyclodextrin.

The medicinal ingredient, for example, vitamin A oil, vitamin A such as retinol, vitamin B ₂ such as riboflavin, B ₆ such as pyridoxine hydrochloride, L- ascorbic acid, L- ascorbic acid phosphoric acid ester, L- Vitamin Cs such as ascorbic acid monopalmitate, L-ascorbic acid dipalmitate, L-ascorbic acid-2-glucoside, pantothenic acids such as calcium pantothenate, vitamin Ds such as vitamin D ₂ and cholecalciferol Vitamins such as vitamin E such as α-tocopherol, tocopherol acetate, DL-α-tocopherol nicotinate; Whitening agents such as placenta extract, glutathione, and Yukinosita extract, skin activators such as royal jelly, beech extract, capsaicin, gingeron, cantalis tincture, ictamol, caffeine, tannic acid, blood circulation promoters such as γ-oryzanol, glycyrrhizic acid Derivatives, glycyrrhetinic acid derivatives, anti-inflammatory agents such as azulene, amino acids such as arginine, serine, leucine and tryptophan, maltose sucrose condensates of resident bacteria control agents, lysozyme chloride and the like. In addition, chamomile extract, parsley extract, beech extract, wine yeast extract, grapefruit extract, honeysuckle extract, rice extract, grape extract, hop extract, rice bran extract, loquat extract, buckwheat extract, yakuinin extract, assembly extract, merirot extract, birch extract, licorice extract , Peony extract, bonito extract, loofah extract, capsicum extract, lemon extract, gentian extract, perilla extract, aloe extract, rosemary extract, sage extract, thyme extract, tea extract, seaweed extract, cucumber extract, clove extract, carrot extract, Examples include various extracts such as maroni extract, hamamelis extract and mulberry extract.

  Cosmetics of the present invention are skin cosmetics such as lotions, emulsions, creams, packs, makeup bases such as makeup base lotions, makeup creams, emulsions or creamy foundations, hand creams, leg creams, body It can be used as body cosmetics such as lotions, bathing agents and the like.

[Example]
Examples 1 to 9 and Comparative Examples 1 to 6 were prepared according to the following procedure with the formulations shown in Tables 1 and 2.
A (Silymarin, surfactant, polyhydric alcohol) and B (strong base, water) in the table were mixed and heated to 70 ° C., B was added to A, mixed uniformly, and cooled to room temperature. The experimental scale was 100 g in total. For Comparative Examples 4 and 5 containing no surfactant or polyhydric alcohol, B at 70 ° C. was directly added to the powder of 70% silymarin-containing Maria thistle extract, mixed uniformly, and cooled to room temperature.
Solubility, stickiness, pH safety, and collagen / elastin production promoting effect were evaluated according to the following criteria, and the results are shown in Tables 1 and 2.

Solubility ◎: Transparent ○: Almost transparent but slightly turbid.
Δ: Translucent ×: Cloudiness

Stickiness ◎: Not sticky ○: Almost non-sticky △: Slightly sticky ×: Sticky

pH safety ○: It is a safe pH when applied to the skin.
(Triangle | delta): It can use safely by setting it as the 2 agent type cosmetics mixed with acidic cosmetics.
X: pH is too high and dangerous.

Collagen / elastin production promoting effect ○: Based on Test Examples 1 to 3, a collagen / elastin production promoting effect can be expected effectively.
Δ: Based on Test Examples 1 to 3, the effectiveness of the collagen / elastin production promoting effect is not high.

As shown in Examples 1 to 3, in a cosmetic containing 0.7% silymarin (70% silymarin-containing maria thistle extract 1%), by adding polyhydric alcohol and a strong base, 5. It could be dissolved with 7 times the amount of surfactant.
As shown in Comparative Example 1, the cosmetic containing 0.7% of silymarin was translucent even when 8.6 times the amount of surfactant was used as silymarin, and silymarin could not be solubilized. Furthermore, since Comparative Example 1 is too sticky, it is not suitable for use as a cosmetic.

Test example 1
[Evaluation of type I collagen production promoting effect in a three-dimensional human skin model]
The human skin three-dimensional model is widely used for safety evaluation and effectiveness evaluation as a pseudo model of human skin. The human skin three-dimensional model was cultured using TESTSKIN (LSE-high) (Toyobo) according to the attached protocol. 70% silymarin-containing Maria thistle extract (Indina), soybean lecithin (Wako Pure Chemicals), and retinoic acid (all-trans-retinoic acid; Wako Pure Chemicals) were dissolved in dipropylene glycol (Wako Pure Chemicals) at each concentration. The 70% silymarin-containing maria thistle extract was dissolved in dipropylene glycol in terms of the concentration of silymarin as an active ingredient. The 70% silymarin-containing Maria thistle extract and soybean lecithin were dissolved in dipropylene glycol by heat treatment, and retinoic acid was dissolved by stirring at room temperature. Dipropylene glycol was used as an untreated control.

  60 μl of each test substance was added onto the tissue in the assay ring and cultured for 24 hours. Thereafter, the culture medium was changed, and the assay ring was washed with a medium, and a new sample was added, followed by further incubation for 24 hours. The tissue was collected, and a tissue extraction solution {50 mM Tris-HCl (pH 7.5), 0.5% (Octylphenoxy) polyethylethanol (Sigma-Aldrich)} was added and homogenized with a Teflon homogenizer. The tissue piece was removed by centrifugation at 10,000 × G for 30 minutes, dialyzed overnight at 4 ° C. in distilled water, and then water was removed by lyophilization. 20 mM Tris-HCl (pH 7.5) was added so as to be 20-fold concentrated, and used as a sample for Western blotting. The amount of protein in the sample was quantified by a Lowry method (Non-patent Document 11) using a DC protein assay kit (Bio-Rad Laboratories), and 20 mM Tris-HCl (pH 7.5) was adjusted to 2 μg / μl. ) To adjust. Add an equal amount of 2x concentrated Laemmli sample buttsfer (Bio-Rad Laboratories) containing 5% 2-mercaptoethanol (Bio-Rad Laboratories) to the sample, and heat-treat at 100 ° C for 5 minutes. Reduced protein.

  Using this sample, the type I collagen production promoting effect was evaluated by Western blotting. 10 μg of protein was applied per lane, separated by SDS-PAGE, and transferred to a nitrocellulose membrane. The transferred nitrocellulose membrane is immersed in a blocking solution (a solution in which skim milk is dissolved in PBS containing 0.1% polyoxyethylene (20) sorbitan monolaurate to a concentration of 5%) at 4 ° C. all day and night. Blocked. After washing with a washing solution {PBS containing 0.1% polyoxyethylene (20) sorbitan monolaurate}, soaked in a primary antibody {polyclonal antibody against type I collagen (Rockland) prepared to 500 ng / ml with the washing solution} The reaction was allowed to proceed for 1 hour at room temperature. After washing, it was immersed in a secondary antibody (horseradish peroxidase-labeled anti-rabbit immunoglobulin G prepared to 250 ng / ml with a washing solution) and allowed to react at room temperature for 1 hour. After washing, detection was performed using an ECL plus western plotting detection reagent (Amersham Bioscience). In addition, the detected band was captured using an image scanner (Amersham Bioscience), and then analyzed and digitized using image master software (Amersham Bioscience).

  The measurement result of the type I collagen production promoting action of each test substance by Western blotting is shown in FIG. Table 3 shows the results of quantification of full-length type I collagen (the band labeled as collagen in FIG. 1).

Table 3

  When treated with 0.5% silymarin, it was equivalent to the untreated control, but it was 2.7 times at 0.7%, 3.3 times at 1.0%, and 2.4 times at 1.5%. In 2.0%, it was promoted 2.2 times. As a positive control, treatment with retinoic acid at 0.05% promoted 2.0 times compared to the untreated control. On the other hand, when soy lecithin was treated with 1.0% as a negative control, it was similar to the untreated control. From this, it became clear that treatment of silymarin with 0.7% or more promotes type I collagen production of the three-dimensional skin model by 2.0 times or more.

Test example 2
[Evaluation of elastin production promoting effect in a three-dimensional human skin model]
The elastin production promoting action in the human skin three-dimensional model was evaluated in the same manner as the evaluation of the type I collagen production promoting action in the three-dimensional human skin model described above.
The measurement results of the elastin production promoting action of each compound by Western blotting are shown in FIG. Table 4 shows the results of quantification of full-length elastin (the band labeled as elastin in FIG. 2).

Table 4

  When treated with 0.5% silymarin, it was equivalent to the untreated control, but 2.5 times at 0.7%, 4.3 times at 1.0%, 3.3 times at 1.5% 2.0%, it was promoted 2.0 times. As a positive control, treatment with retinoic acid at 0.05% promoted 3.3 times compared to the untreated control. On the other hand, when soy lecithin was treated with 1.0% as a negative control, it was similar to the untreated control. From this, it became clear that the elastin production of a three-dimensional skin model was promoted 2.0 times or more by treating silymarin with 0.7% or more.

Test example 3
[Evaluation of percutaneous absorption of silymarin using pig skin]
We evaluated the percutaneous absorption of silymarin using pig skin, which is reported to be close to human skin absorption. The evaluation was performed by a method using a diffusion cell devised by Franz. Yucatan minipig skin (five months old female; Nippon Charles River) stored at −80 ° C. after collection was allowed to stand at room temperature for about 20 minutes to be in a half-thawed state. Fat was removed with both blunt scissors for surgery. Put the phosphate buffer physiological saline solution (pH 7.4) containing 0.05% kanamycin sulfate (Wako Pure Chemical Industries) in the receptor cell in the lower layer of the diffusion cell devised by Franz, set the pig skin, and set the upper donor cell did. The cell was placed in a 37 ° C. incubator, and the receptor cell solution was stirred with a stirrer. 100 μl of a sample obtained by dissolving 70% silymarin-containing maria thistle extract in dipropylene glycol at each silymarin concentration was added to the donor cell and sealed with parafilm. Each silymarin concentration was evaluated using two pig skins. After 24 hours, the sample remaining on the pig skin was removed, and only the skin where the sample was applied was removed with scissors. The skin was sandwiched with a spatula warmed in an 80 ° C. water bath, heated for 20 seconds, and the epidermis and dermis were peeled off using tweezers. The dermis was finely chopped with scissors and then placed in a 15 ml centrifuge tube. 2 ml of 99.8% methanol (Wako Pure Chemical Industries, Ltd.) was added and crushed with a polytron homogenizer. The tissue piece was removed by centrifugation at 10,000 × G for 30 minutes, and then filtered through a 0.45 μm membrane filter to obtain a sample for component analysis.
Silymarin infiltrated into the dermis of pig skin was measured by high performance liquid chromatography using silymarin in a sample prepared by dissolving 70% silymarin-containing Maria thistle extract in dipropylene glycol at each silymarin concentration. The conditions for high performance liquid chromatography are shown below. When analyzed under the following conditions, peaks of silicristin, silidinin, silibinin A, silibinin B, isociribine A, and isociribine B are observed.

(High performance liquid chromatography conditions)
Column; C18 UG120 4.6mm-250mm (Shiseido)
Mobile phase; water: methanol: acetic acid = 65: 35: 5
Column temperature: 40 ° C
Measurement wavelength: 288 nm
Flow rate: 1.0 ml / min

The total value of the peak areas of each component was calculated as the silymarin concentration, and the amount of silymarin in a sample obtained by dissolving 70% silymarin-containing maria thistle extract in dipropylene glycol at each silymarin concentration was taken as 100% and penetrated into the dermis of pig skin. The amount of silymarin was expressed in%. The results were represented by graphing the average value of silymarin concentration penetrating into the dermis of two pig skins (FIG. 3). The slope of 0.7% to 2.0% is moderate with respect to the slope of silymarin concentration of 0.5% to 0.7%, and the permeability to the dermis is rapidly increasing at 0.7%. It has been shown.
Silymarin exhibits type I collagen and elastin production promotion effect in human skin based on the evaluation of silymarin's type I collagen and elastin production promotion effect on three-dimensional skin models and the evaluation of silymarin penetration into porcine skin Therefore, a silymarin concentration of 0.7% or more was considered effective.

The figure which shows the production promotion effect | action of type I collagen in the human three-dimensional skin model by a silymarin. The figure which shows the production promotion effect of the elastin in the human three-dimensional skin model by a silymarin. The figure which shows the permeability | transmittance to the dermis of the pig skin of silymarin.

Claims (5)

A cosmetic comprising the following (1) to (5) and not containing ethanol .
(1) Silymarin (2) One or more polyhydric alcohols selected from polyethylene glycol, dipropylene glycol, 1,3-butylene glycol, 1,2-pentanediol, propylene glycol (3) For the amount of silymarin 0.5 to 6 times the amount of the surfactant , the surfactant is polyoxyethylene hydrogenated castor oil and / or polyoxyethylene polyoxypropylene alkyl ether,
(4) Strong base (5) Water The cosmetic according to claim 1, wherein the concentration of silymarin is 0.7% by mass or more. The cosmetic according to claim 1 or 2, which has a pH of 6.0 to 9.5. The cosmetic according to any one of claims 1 to 3, which has a pH of 7.5 to 9.5. A two-pack type cosmetic, wherein the cosmetic according to claim 4 and an acidic cosmetic having a pH of 3.0 to 6.0 are separately filled in a container and mixed when used.