JP6611308B2 - Skin preparation - Google Patents

Description

  The present invention relates to a skin external preparation having an excellent anti-aging action.

  Research on so-called skin aging suppression, such as prevention of skin wrinkle formation and prevention of skin sagging, is underway. Among them, fibroblasts are important cells that produce substances such as collagen and elastin that function to maintain skin tension and elasticity, and thus various studies have been conducted.

Versican known to be produced by fibroblasts is a large chondroitin sulfate proteoglycan and is known to be temporarily expressed at a high level during wound healing (Non-patent Document 1).
Versican is also known to play an important role in the creation of the skin of fetuses and newborns. Specifically, collagen and other components produced by fibroblasts with versican as a scaffold are three-dimensional in the dermis. Assist in forming the structure (Non-Patent Document 2). It is also known that versican disappears after the dermis structure is completed, and in adults it is hardly expressed except during wound healing as described above (Non-patent Document 3).
It has also been reported that versican can carry hyaluronic acid (Non-Patent Document 4).

  On the other hand, silk, especially hydrolyzed silk protein, has been reported to have various skin effects, such as antioxidant action, skin inflammation prevention action, tyrosinase activity inhibition action, collagen production promotion action, moisture retention improvement action, wrinkle formation An inhibitory action is mentioned (patent documents 1, 2).

  In addition, lupeol, which is one of the triterpenoids, is known to be contained in various plant extracts, and it has been reported that lupeol and its derivatives have effects such as suppression of melanin production (Patent Literature). 3-5).

JP 2007-277119 A JP 2005-15446 A JP 2007-511472 A JP 2004-345959 A Japanese Patent Laid-Open No. 2002-3381

ARAKI Eri et al., Abstracts of Annual Meeting of the Connective Tissue Society of Japan 41, 54 Sorrell JM. Et al., Anat. Embryol., 1999, 199 (1) 45-56 Carrino DA. Et al., Arch Biochem. Biophys., 2000, 373 (1) 91-101 Matsumoto K. et al., J. Biol. Chem., 2003, 278 (42) 41205-41212

  This invention makes it a subject to provide the skin external preparation which has the outstanding anti-aging effect.

  The inventors of the present invention have found that an extract extracted by a specific method from golden silk having a dark yellow amber color among silks promotes versican production in fibroblasts (Japanese Patent Application 2014). -231340). As a result of further diligent research to solve the above problems, it was found that lupeol is an effective component for promoting versican production contained in the extract. Furthermore, it has been conceived that by combining lupeol with hyaluronic acid and collagen, it is possible to improve the moisture retention ability and elasticity of the skin and to provide a skin external preparation that exhibits an excellent anti-aging action.

That is, one aspect of the present invention is a skin external preparation containing lupeol and hyaluronic acid.
Moreover, another aspect of the present invention is an external preparation for skin containing a golden silk extract and hyaluronic acid. In this embodiment, the golden silk extract is preferably obtained by extracting from golden silk with an acidic alcohol solvent of 60 ° C. or higher or an acidic mixed solvent of alcohol and water.
Furthermore, it is preferable that the skin external preparation of this invention contains collagen further.
Furthermore, the skin external preparation of the present invention is preferably a cosmetic, and more preferably an anti-aging cosmetic.

The skin external preparation which exhibits the outstanding anti-aging effect is provided by this invention.
The external preparation for skin of the present invention promotes versican production by fibroblasts, and can create an environment similar to that when the dermis is created in a fetus or a newborn. And since the produced versican carries hyaluronic acid, the moisture retention ability of the skin can be improved. Further, as a result of promoting versican production, a thick and tough three-dimensional structure of collagen fiber bundles can be formed, and the elasticity of the skin can be improved. As a result, the external preparation for skin of the present invention can provide a dermis that is rich and elastic and freshly reborn, and thus can provide an excellent anti-aging cosmetic.

The graph which shows the expression level of versican mRNA in the reference test example 2. FIG. ¹ H-NMR chart of golden silk extract fraction and lupeol. ¹³ C-NMR chart of golden silk extract fraction and lupeol. 10 is a photomicrograph showing the distribution of versican in Reference Test Example 3. 9 is a micrograph showing a collagen fiber bundle in Reference Test Example 4. The graph which shows the elasticity value of the collagen gel in the reference test example 4. FIG. The graph which shows the water retention ability of the collagen gel in the reference test example 5. FIG. 6 is a photograph showing the appearance of a collagen gel in Reference Test Example 5.

  Hereinafter, the present invention will be described in detail, but it is needless to say that the present invention is not limited to specific embodiments.

The skin external preparation of the present invention contains lupeol.
Lupeol is a lupine-type triterpene (CAS number: 545-47-1, molecular weight 426.724). It is known that it is contained in various plants in nature.
The present inventors previously found that the golden silk extract has an excellent versican production-promoting action, and found that its active ingredient is lupeol. Lupeol is thought to be also contained in mulberry leaves, but lupeol also exists in the cocoons made by cocoons that ate mulberry leaves.
Lupeol can be obtained with high purity by extracting from straw. In particular, when extracted from cocoons called golden silk, lupeol with higher purity can be obtained.
Therefore, in the other aspect of this invention, a skin external preparation contains a golden silk extract.

The lupeol according to the present invention may be used after being purified from a golden silk extract or may be used as a golden silk extract.
The golden silk extract may be in the form of an extract, oil, or powder, and the extract extracted as an extract or oil using a solvent may be dried to obtain a powdery extract.

Golden Silk (Golden Silk No. 11) is silkworm (Bonbix)
mori), which is dark yellow. It is a special paddy produced mainly in Thunghuachang Lamphun Thailand, Lampung Province, Kingdom of Thailand, and is also commercially available.

  The golden silk extract in the present invention can be obtained by any production method, but it can be obtained by a production method including a step of extracting golden silk with an acidic alcohol solvent of 60 ° C. or higher or an acidic mixed solvent of alcohol and water. However, it is preferable because an extract containing a large amount of lupeol and having a high ability to promote versican production can be obtained.

  The alcohol solvent is preferably a lower alcohol, and examples thereof include methanol, ethanol, propyl alcohol, and isopropyl alcohol. Among these, methanol and ethanol are preferable, and ethanol is more preferable from the viewpoint of extraction efficiency. Moreover, in the mixed solvent of alcohol and water, it is usually preferable that the alcohol is contained in an amount of 40% v / v or more, particularly preferably 50% v / v or more from the viewpoint of extraction efficiency. Moreover, an upper limit is not specifically limited, Usually, it is 99.9% v / v or less.

The reason for making the extraction solvent acidic is to hydrolyze golden silk, and the pH is usually less than 7 and preferably 3 or less, more preferably 2 or less. The lower limit of the pH is not particularly limited, but is usually 1 or more.
Although golden silk can be hydrolyzed with an alkaline solvent, the golden silk extract extracted with an alkaline solvent is strongly yellowish-brown and is not preferable when blended with a composition such as a skin external preparation. On the other hand, the golden silk extract extracted with an acidic solvent is less colored.
In order to make the extraction solvent acidic, for example, an appropriate amount of acid such as sulfuric acid, hydrochloric acid, or nitric acid can be added to the solvent. Of these, hydrochloric acid is particularly preferred because the extract can be less colored.

Extraction is carried out at 60 ° C. or higher, preferably 80 ° C. or higher. The upper limit of the heating is not particularly limited, and may be performed within a range in which safety can be ensured without excessively changing the pressure.
Although extraction depends on the extraction temperature, it is usually performed for 10 minutes or longer, more preferably 30 minutes or longer, and further preferably for 1 hour or longer. Although the upper limit of extraction time is not specifically limited, Usually, it is 20 hours or less, and it is preferable that it is 10 hours or less.

The obtained extract is preferably neutralized with an alkali. The neutralization can be performed by adding an alkali such as NaOH or KOH.
In addition, it is possible to increase the purity by precipitating starch as appropriate, or to remove impurities by providing a filtration step.

  The method for obtaining lupeol from the golden silk extract can be performed by a conventional method.

In the external preparation for skin of the present invention, the content of lupeol is not particularly limited, but is usually preferably 0.00001 to 0.05 mass%, and 0.0001 to 0.05 mass% with respect to the entire external preparation for skin. More preferably, 0.001-0.05 mass% is further more preferable.
Further, in the external preparation for skin of the present invention, the content of the golden silk extract is not particularly limited, but usually 0.0001 to 0.5% by mass is preferable as an evaporation residue with respect to the entire external preparation for skin. 0.001 to 0.5 mass% is more preferable, and 0.01 to 0.5 mass% is more preferable.

  As described above, versican is usually hardly expressed in adults. In the external preparation for skin of the present invention, lupeol or golden silk extract can promote versican production by fibroblasts and compensate for versican deficiency in adults. As a result, it promotes the formation of a three-dimensional structure of an extracellular matrix such as a collagen fiber bundle, and as a result, plays a role leading to the creation (regeneration) of the dermis.

The skin external preparation of the present invention also contains hyaluronic acid.
Hyaluronic acid is a polysaccharide generally known as a water retention component. In the external preparation for skin of the present invention, hyaluronic acid is held in versican whose production is promoted by lupeol or golden silk extract, and plays a role of improving water retention ability in the dermis by the water retention ability of hyalurone itself.

In the external preparation for skin of the present invention, the content of hyaluronic acid is not particularly limited, but is usually preferably 0.0001 to 0.5 mass%, and preferably 0.001 to 0.5 mass% with respect to the entire external preparation for skin. Is more preferable, and 0.01-0.5 mass% is further more preferable.
The average molecular weight of hyaluronic acid is not particularly limited, but is usually preferably less than 500,000, more preferably less than 10,000, and even more preferably less than 1,000.

In the external preparation for skin of the present invention, lupeol and hyaluronic acid are preferably contained at a mass ratio of 1: 1, more preferably at a mass ratio of 1:10.
Moreover, in the skin external preparation of this invention, it is preferable to contain lupeol and golden silk extract (evaporation residue) by the mass ratio of 1: 1, and it is more preferable to contain by the mass ratio of 1: 5.

The external preparation for skin of the present invention preferably further contains collagen.
Over 90% of the dermis that functions as a support for the skin structure and plays a mechanical role is composed of collagen. It is known that collagen forms a fiber bundle in the dermis and entangles with almost all layers of the dermis to form a three-dimensional network structure, contributing to skin elasticity. In the external preparation for skin of the present invention, collagen forms a thicker and stronger fiber bundle than usual by the action of versican whose production is promoted by lupeol or golden silk extract, and plays a role of improving the elasticity of the dermis.

In the external preparation for skin of the present invention, the content of collagen is not particularly limited, but is usually preferably 0.0001 to 0.5 mass%, and preferably 0.001 to 0.5 mass% with respect to the entire external preparation for skin. More preferred is 0.01 to 0.5% by mass.
The average molecular weight of collagen is not particularly limited, but is usually preferably less than 300000, more preferably less than 5000, and still more preferably less than 1000.

  The skin external preparation of the present invention can be applied to any application such as cosmetics and quasi drugs, but is preferably a cosmetic, and particularly preferably an anti-aging cosmetic.

  In addition to the above, the skin external preparation of the present invention can optionally contain components such as active ingredients usually used in skin external preparations. Also, the dosage form is not particularly limited and may be arbitrarily designed, such as a lotion dosage form, an emulsifier type such as a milky lotion or cream, and an oil dosage form.

Examples of other active ingredients contained in the external preparation for skin of the present invention include whitening ingredients, wrinkle-improving ingredients, anti-inflammatory ingredients, extracts derived from animals and plants, and the like. It may be contained. Moreover, the content is good also as an amount normally contained in a skin external preparation.
In addition, oily components, surfactants, polyhydric alcohols, thickeners, powders, ultraviolet absorbers and the like can be mentioned as optional components.

  The whitening component is not particularly limited. For example, whitening components such as ascorbic acids, hydrogen peroxide, colloidal sulfur, glutathione, hydroquinone, catechol, tyrosinase-related protein inhibitor, endothelin action inhibitor, proton pump inhibitor, stem cell factor Examples include melanin production inhibitors such as binding inhibitors, dendrite elongation inhibitors, melanin transfer or release inhibitors, integrin production promoters, and the like.

  The wrinkle-improving component is not particularly limited. For example, vitamin A or a derivative thereof, retinol, retinal, retinoic acid, tretinoin, isotretinoin, retinoic acid tocopherol, retinol palmitate, retinol acetate, benzyl ursolate, phosphorus ursolate Examples include acid esters, betulinic acid benzyl ester, and benzyl acid phosphate.

  Examples of the anti-inflammatory component include, but are not limited to, for example, clarinone, glabrizine, glycyrrhizic acid, glycyrrhetinic acid, pantothenyl alcohol, and the like. Examples include acids and salts thereof.

The extracts derived from animals and plants are not particularly limited. Mushroom extract, Fennel extract, Udo extract, Ages extract, Ezocogi extract, Enmeiso extract, Ogon extract, Oat extract, Auren extract, Panax ginseng extract, Hypericum extract, Adrianthus extract, Orange extract, Oyster extract, Cuckoo extract, Chamomile extract, Carrot extract , Kawaramugi Extract, Licorice Extract, Kiwi Extract, Cucumber Extract, Guava Extract, Kujin Extract, Gardenia Extract, Kumazasa Extract, Clara Echi , Walnut extract, grapefruit extract, black rice extract, chlorella extract, mulberry extract, quette extract, gentian extract, gentian extract, gentian pepper extract, tea extract, burdock extract, rice extract, rice fermented extract, rice bran fermented extract, rice germ oil, cowberry Extract, Salvia extract, Soap extract, Sasa extract, Hawthorn extract, Sansha extract, Salamander extract, Shiitake extract, Giant extract, Shikon extract, Perilla extract, Linden extract, Citrus extract, Peonies extract, Ginger extract, Ginger root extract, Birch extract, Horsetail extract, Stevia extract, Stevia fermented product, Kizuta extract, Hawthorn extract, Elderberry extract, Achillea millefolium extract Mint extract, sage extract, mallow extract, senkyu extract, senbu extract, scorpion extract, soybean extract, soy extract, tiso extract, thyme extract, dandelion extract, tea extract, clove extract, chimpi extract, strawberry tea extract, red pepper extract, red pepper extract, Tokachinenka extract, tonin extract, spruce extract, dokudami extract, tomato extract, natto extract, carrot extract, garlic extract, wild rose extract, hibiscus extract, bacmond extract, lotus extract, parsley extract, birch extract, clam extract, cypress extract, cypress extract , Loquat extract, Japanese dandelion extract, Japanese quince extract, Bukuryo extract, Butcher bloom extract, grape extract, grape seed extract, loofah Ma extract, safflower extract, peppermint extract, body extract, button extract, hop extract, pine extract, maroonni extract, mizuba sho extract, mukuroji extract, melissa extract, mozuku extract, peach extract, cornflower extract, eucalyptus extract, yukinoshita extract, yuzu extract, lily extract, yokoinin Extract, mugwort extract, lavender extract, green tea extract, apple extract, rooibos tea extract, litchi extract, lettuce extract, lemon extract, forsythia extract, forsythia extract, rose extract, rosemary extract, roman chamomile extract, royal jelly extract, bitumen extract, etc. Is mentioned.

Examples of the oil component include polar oil and volatile hydrocarbon oil.
Polar oils include synthetic ester oils such as isopropyl myristate, cetyl octanoate, octyldodecyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, decyl oleate, hexyldecyl dimethyloctanoate, lactic acid Cetyl, myristyl lactate, lanolin acetate, isocetyl stearate, isocetyl isostearate, cholesteryl 12-hydroxystearylate, ethylene glycol di-2-ethylhexylate, dipentaerythritol fatty acid ester, N-alkylglycol monoisostearate, neopentyl dicaprate Glycol, diisostearyl malate, glycerin di-2-heptylundecanoate, trimethylolpropane tri-2-ethylhexylate It may be mentioned trimethylolpropane triisostearate, tetra-2-ethylhexyl acid pentane erythritol, tri-2-ethylhexyl acid glycerin, trimethylolpropane triisostearate.

  Further, cetyl 2-ethylhexanoate, 2-ethylhexyl palmitate, glyceryl trimyristate, glyceride tri-2-heptylundecanoate, castor oil fatty acid methyl ester, oleic acid oil, cetostearyl alcohol, acetoglyceride, palmitic acid 2 -Heptylundecyl, diisobutyl adipate, N-lauroyl-L-glutamic acid-2-octyldodecyl ester, di-2-heptylundecyl adipate, ethyl laurate, di-2-ethylhexyl sebacate, 2-hexyl myristate Decyl, 2-hexyldecyl palmitate, 2-hexyldecyl adipate, diisopropyl sebacate, 2-ethylhexyl succinate, ethyl acetate, butyl acetate, amyl acetate, triethyl citrate, octyl Toki Shishi runner formate may also be mentioned.

  Natural oils include avocado oil, camellia oil, turtle oil, macadamia nut oil, corn oil, mink oil, olive oil, rapeseed oil, egg yolk oil, sesame oil, persic oil, wheat germ oil, sasanqua oil, castor oil, flaxseed oil, Safflower oil, cottonseed oil, eno oil, soybean oil, peanut oil, tea seed oil, kaya oil, rice bran oil, cinnagari oil, Japanese kiri oil, jojoba oil, germ oil, triglycerin, trioctanoic acid glycerin, triisopalmitic acid glycerin Etc.

  Examples of the volatile hydrocarbon oil include isododecane and isohexadecane.

As surfactants, fatty acid soap (sodium laurate, sodium palmitate, etc.), anionic surfactants such as potassium lauryl sulfate, alkylsulfuric triethanolamine ether, stearyltrimethylammonium chloride, benzalkonium chloride, laurylamine oxide Cationic surfactants such as
Betaine surfactants (alkyl betaines, amide betaines, sulfobetaines, etc.), imidazoline amphoteric surfactants (such as 2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt), acylmethyl taurine Amphoteric surfactants such as
Sorbitan fatty acid esters (such as sorbitan monostearate and sorbitan sesquioleate), glycerin fatty acids (such as glyceryl monostearate), propylene glycol fatty acid esters (such as propylene glycol monostearate), hydrogenated castor oil derivatives, glycerin alkyl ether POE sorbitan fatty acid esters (POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbite fatty acid esters (POE-sorbitol monolaurate, etc.), POE glycerin fatty acid esters (POE-glycerin monoisosteare) Rate), POE fatty acid esters (polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (POE2-octyl) Decyl ether, etc.), POE alkylphenyl ethers (POE nonylphenyl ether, etc.), pluronic types, POE / POP alkyl ethers (POE / POP2-decyltetradecyl ether, etc.), tetronics, POE castor oil / cured castor Examples include oil derivatives (POE castor oil, POE hydrogenated castor oil, etc.), nonionic surfactants such as sucrose fatty acid esters, alkyl glucosides, and the like.

  Polyhydric alcohols include polyethylene glycol, glycerin, 1,3-butylene glycol, erythritol, sorbitol, xylitol, maltitol, propylene glycol, dipropylene glycol, diglycerin, isoprene glycol, 1,2-pentanediol, 2,4 -Hexylene glycol, 1,2-hexanediol, 1,2-octanediol and the like.

  Examples of thickeners include guar gum, quince seed, carrageenan, galactan, gum arabic, pectin, mannan, starch, xanthan gum, curdlan, methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, methylhydroxypropylcellulose, chondroitin sulfate, dermatan sulfate, glycogen, Heparan sulfate, hyaluronic acid, sodium hyaluronate, tragacanth gum, keratan sulfate, chondroitin, mucoitin sulfate, hydroxyethyl guar gum, carboxymethyl guar gum, dextran, kerato sulfate, locust bean gum, succinoglucan, caronic acid, chitin, chitosan, carboxymethyl Chitin, agar, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, al Le-modified carboxyvinyl polymers, sodium polyacrylate, polyethylene glycol, and bentonite.

  As powders, the surface may be treated, mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, silicic anhydride (silica), aluminum oxide, barium sulfate, etc. May be treated, bengara, yellow iron oxide, black iron oxide, cobalt oxide, ultramarine, bitumen, titanium oxide, zinc oxide inorganic pigments, surface may be treated, titanium mica, fish phosphorus foil, Pearl agent such as bismuth oxychloride, red 202, red 228, red 226, yellow 4, blue 404, yellow 5, red 505, red 230, red 223 which may be raked No., Orange No. 201, Red No. 213, Yellow No. 204, Yellow No. 203, Blue No. 1, Green No. 201, Purple No. 201, Red No. 204, organic dyes, polyethylene powder, polymeta Methyl acrylic acid, nylon powder, organic powders such as organopolysiloxane elastomers, and the like.

Examples of the UV absorber include paraaminobenzoic acid UV absorbers, anthranilic acid UV absorbers, salicylic acid UV absorbers, cinnamic acid UV absorbers, benzophenone UV absorbers, sugar UV absorbers, 2- (2 UV absorbers such as'-hydroxy-5'-t-octylphenyl) benzotriazole, 4-methoxy-4'-t-butyldibenzoylmethane, and the like.

  Hereinafter, the present invention will be described in more detail with specific experimental examples. However, the present invention is not limited to the following embodiments.

<Reference Test Example 1>
10 g of golden silk (Golden Silk No. 11 from Tonhuachachan, Lampung, Thailand) was immersed in a mixed solvent of 400 mL of 50% ethanol solution and 1 mL of concentrated hydrochloric acid, and extracted at 80 ° C. for 7 hours. After extraction, the filtered filtrate was neutralized with 1N NaOH. Thereafter, the neutralized extract was allowed to stand at 5 ° C. to precipitate starch. The starch was filtered off to obtain a golden silk extract.
Human fibroblasts were seeded at 7.0 × 10 ⁵ cells / well in 10% FBS / DMEM medium, and cultured at 37 ° C. under 5% CO ₂ conditions. After 24 hours from the start of culture, the medium was removed and the cells were
After washing with BS (−), the golden silk extract was added as an evaporation residue to a concentration of 1 × 10 ⁻³ mass% in 2% FBS / DMEM medium, and further cultured for 48 hours. After the culture, mRNA was collected, and the expression level of versican mRNA was measured by RT-PCR. The expression level was calculated based on the internal standard substance. Also, when 50% ethanol is added instead of the extract as a negative control (control), and when 2 ng / mL of TGF-β having anti-aging action is added as a positive control, the mRNA expression level is measured in the same manner. did.
The relative expression level when the negative control (control) is 1 is shown in FIG. As can be seen from this result, the expression of versican in fibroblasts was promoted by the golden silk extract.

<Reference Test Example 2>
20 g of golden silk was immersed in a mixed solvent of 400 mL of methanol, 400 mL of water and 2 mL of concentrated hydrochloric acid, and extracted at 80 ° C. for 6 hours. After extraction, the filtered filtrate was neutralized with 1N NaOH. Thereafter, the neutralized extract was allowed to stand at 5 ° C. for 1 day or longer to precipitate starch. The starch was filtered off to obtain a golden silk extract.
After the solvent was distilled off from the above golden silk extract, ethyl acetate was added to perform liquid-liquid partitioning. The ethyl acetate layer was dried and purified with a silica gel column using chloroform as a solvent.
¹ H-NMR, ¹³ C-NMR, and MS analysis were performed on the fraction that promoted the expression of versican in fibroblasts by performing the same experiment as in Reference Test Example 1, and these were lupeol (reagent) (Figs. 2-3) and the literature values (EXCLI Journal 2010; 9: 1-10, Research Journal of Pharmaceutical Sciences, Vol. 1 (1), 23-27, 2012) Lupeol was identified as the active ingredient of the golden silk extract for promoting versican production.

<Reference Test Example 3>
Human fibroblasts are seeded at 1.5 × 10 ⁴ cells / well on a collagen gel, and a golden silk extract is added as an evaporation residue so as to have a concentration of 1 × 10 ⁻³ mass%. The cells were cultured for 10 days according to the gel culture method. As a control, human fibroblasts without addition of golden silk extract were also cultured in the same manner. Anti-versican antibody (R & D system: Human Versican Isoform VO Antibody)
s) and observed with a confocal fluorescence microscope (LSM510 Laser Module: ZEISS).
The results are shown in FIG. It can be seen that the golden silk extract promotes versican production in fibroblasts and the collagen gel is filled with versican.

<Reference Test Example 4>
Human fibroblasts are seeded at 1 × 10 ⁶ cells / well on a collagen gel, and a golden silk extract is added as an evaporation residue so as to have a concentration of 1 × 10 ⁻³ mass%. According to the method, it was cultured for 5 days. As a control, human fibroblasts without addition of golden silk extract were also cultured in the same manner. After culturing, collagen fiber bundles were observed with a scanning electron microscope (VSM-6380LA: JEOL). Moreover, the elasticity value of the collagen gel after culture | cultivation was measured using the baristometer (Keystone Scientific Co., Ltd.).
From the results shown in FIG. 5, it can be seen that when the golden silk extract was added, many thicker and stronger collagen fiber bundles were formed. Moreover, as shown in FIG. 6, when a golden silk extract is added, it turns out that the elasticity of a collagen gel improved about 1.5 times.

<Reference Test Example 5>
Human fibroblasts are seeded at 1 × 10 ⁵ cells / well on a collagen gel, and golden silk extract is added as an evaporation residue to a concentration of 1 × 10 ⁻³ mass%, and normal collagen gel culture is performed. According to the method, it was cultured for 6 days. As a control, human fibroblasts without addition of golden silk extract were also cultured in the same manner. After the culture, the collagen gel was immersed in a 0.25% by mass hyaluronic acid aqueous solution for 30 minutes, and the amount of increase in water was measured after the immersion. Thereafter, the collagen gel was dried (37 ° C., 2 hours), and the appearance of the collagen gel before and after the drying treatment was observed.
From the results shown in FIG. 7, it can be seen that when the golden silk extract was added, the water retention capacity of the collagen gel was improved by about 1.2 times. In addition, as shown in FIG. 8, the collagen gel after the drying treatment released water in the control and became transparent, whereas when the golden silk extract was added, the gel might still contain water. Understand.

<Example 1>
The skin external preparation of this invention was manufactured by the prescription shown below. That is, the prescription ingredients were heated, stirred, solubilized at 70 ° C., stirred and cooled to obtain the lotion of Example 1.
Formula:
Lupeol 0.1 part by weight Hyaluronic acid 0.1 part by weight Glycerin 5 part by weight 1,2-pentanediol 4 part by weight Polyphosphatidyloxyethyl methacrylate 0.1 part by weight Phenoxyethanol 0.1 part by weight Ethanol 5 part by weight POE (60) Hardened castor oil 0.1 parts by weight Water 85.5 parts by weight

  Further, the lotion of Comparative Example 1 in which hyaluronic acid was replaced with water in the lotion of Example 1, the lotion of Comparative Example 2 in which lupeol was replaced with water in the lotion of Example 1, and the lotion of Example 1 The skin lotion of Comparative Example 3 in which lupeol and hyaluronic acid were replaced with water was also prepared in the same manner.

  With respect to the lotions of Example 1 and Comparative Examples 1 to 3, a long-term continuous test was conducted using a panel of 10 people per group who suffered from elasticity, elasticity, and freshness. Each group of panelists applied lotion to the face once a day every day. Three months after the start of the test, the improvement of elasticity, elasticity, and freshness were marked, effective, and ineffective, respectively. We had you evaluate in stage. From the results shown in Table 1, it can be seen that the cosmetic according to the present invention exhibits an excellent improvement effect on elasticity, elasticity, and freshness.

<Example 2>
The skin external preparation of this invention was manufactured by the prescription shown below. That is, the components of A, B and C were heated to 70 ° C., respectively, and B was added to neutralize, and C was gradually added to emulsify. This was stirred and cooled to obtain the emulsion of Example 2.
Formula:
(I)
1,2-pentanediol 5 parts by weight Isoprene glycol 4 parts by weight Acrylic acid / alkyl methacrylate polymer 0.6 parts by weight Phenoxyethanol 0.1 part by weight Water 30 parts by weight (b)
10% aqueous potassium hydroxide solution 3 parts by weight water 39.7 parts by weight (c)
Squalane 5 parts by weight Jojoba oil 5 parts by weight Olive oil 5 parts by weight Golden silk extract extract 0.2 parts by weight Hyaluronic acid 0.2 parts by weight

  In addition, the lotion of Comparative Example 4 in which hyaluronic acid was replaced with water in the emulsion of Example 2, the lotion of Comparative Example 5 in which lupeol was replaced with water in the emulsion of Example 2, and the emulsion lupeol and hyaluron of Example 2 The lotion of Comparative Example 6 in which the acid was replaced with water was also prepared in the same manner.

  With regard to the lotions of Example 1 and Comparative Examples 1 to 3, an actual use test was conducted using a panel of 10 people who suffered from elasticity, elasticity, and freshness. Each group of panelists applied lotion to the face once a day every day, 4 weeks after the start of the test, the improvement of elasticity, elasticity, and freshness was marked, effective, and ineffective 3 We had you evaluate in stage. From the results shown in Table 2, it can be seen that the emulsion according to the present invention exhibits an excellent improvement effect on elasticity, elasticity, and freshness.

  The external preparation for skin of the present invention is rich in elasticity and elasticity, and can realize a fresh and reborn skin. Therefore, it can be an excellent anti-aging cosmetic and is very useful industrially.

Claims (3)

A skin external preparation for improving elasticity, elasticity, and / or freshness, comprising a golden silk extract and hyaluronic acid ,
The said golden silk extract is a skin external preparation obtained by the manufacturing method including the process of extracting golden silk with an acidic alcohol solvent of 60 degreeC or more or the acidic mixed solvent of alcohol and water . Further comprising collagen, skin external preparation according to claim 1. The external preparation for skin according to claim 1 or 2 , which is a cosmetic.