JP2015131797A - Versican production promoter, and method for producing golden silk extract - Google Patents

Abstract

A novel versican production promoter is provided. A method for producing a golden silk extract comprising the step of extracting golden silk with a solvent at 60 ° C. or higher. The method for producing a golden silk extract, wherein the solvent is an acidic alcohol solvent or an acidic solvent of alcohol and water. A versican production promoter comprising a golden silk extract produced by the production method. Furthermore, the skin external preparation which contains this versican production promoter, and this skin external preparation which is anti-aging cosmetics. [Selection] Figure 1

Description

  The present invention relates to a versican production promoter involved in anti-aging. Moreover, it is related with the manufacturing method of the golden silk extract excellent in the production promotion effect of versican.

  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 (see Non-Patent Document 1).

  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 Examples thereof include an inhibitory action (see Patent Documents 1 and 2).

JP 2007-277119 A JP 2005-15446 A

ARAKI Eri, et al., Abstracts of Annual Meeting of the Connective Tissue Society of Japan 41, 54

  An object of the present invention is to provide a new versican production promoter and a new method for producing a golden silk extract.

The present inventors have studied to solve the above-mentioned problems, and found that a golden silk extract having a dark yellow amber color among silk promotes versican production in fibroblasts, and completed the present invention. It was.
In addition, further research was conducted on the extraction method of the golden silk extract, and it was found that the golden silk extracted by a specific method has a higher versican production promoting effect.

That is, the 1st aspect of this invention is a versican production promoter which consists of a golden silk extract obtained by the specific manufacturing method.
Moreover, the 2nd aspect of this invention is a manufacturing method of the golden silk extract including the extraction step which extracts golden silk with an acidic alcohol solvent or the acidic mixed solvent of alcohol and water at 60 degreeC or more.

According to the present invention, a new versican production promoter is provided. The skin external preparation which exhibits the outstanding anti-aging effect can be provided by including the versican production promoter of this invention in a skin external preparation. Therefore, it is preferably used as an anti-aging cosmetic.
Moreover, according to the second aspect of the present invention, a golden silk extract with enhanced versican production promoting action is provided. The golden silk extract produced by this production method exhibits a high versican production promoting action that exceeds TGF-β, which has been confirmed to have a high anti-aging action. Therefore, it is preferably used as an active ingredient of anti-aging cosmetics.

It is a graph which shows the expression level of versican mRNA in an Example. It is a graph which shows the expression level of versican mRNA in an Example.

  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 golden silk (Golden Silk No. 11) used in the embodiment of the present invention is a silkworm of Bombyx mori and has a deep yellow color. It is a special paddy produced mainly in Thunghuachang Lamphun Thailand, Lampung Province, Kingdom of Thailand, and is also commercially available.

In this embodiment, the 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.
Of the production method examples of the golden silk extract used in the present embodiment, preferred embodiments of the production method of the extract are shown below.
<Extraction step>
The manufacturing method according to this embodiment includes an extraction step of extracting golden silk with a solvent of 60 ° C. or higher. By including this step, a golden silk extract with high versican production ability can be obtained. The solvent is preferably an acidic alcohol solvent or an acidic mixed solvent of alcohol and water.
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.

Moreover, in order to hydrolyze golden silk, it is preferable that the extraction solvent in this embodiment is acidic. 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. Since the golden silk extract extracted with an acidic solvent is less colored, an acidic solvent is used in this embodiment.
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.

The extraction step is performed under heating. By performing the extraction step under heating, a golden silk extract with high versican production ability can be obtained.
Heating is 60 ° C. or higher, preferably 80 ° C. or higher. On the other hand, the upper limit is not particularly limited, and may be set within a range in which safety can be secured without excessively changing the pressure.

  The extraction in the extraction step is usually performed for 10 minutes or more, more preferably 30 minutes or more, and further preferably 1 hour or more, although it depends on the extraction temperature. 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.

In the manufacturing method according to this embodiment, it is preferable that the extract extracted in the extraction step further undergoes the following steps.
<Neutralization step>
When an acidic solvent is used, the extracted extract is acidic due to hydrolysis of golden silk, and therefore preferably has a step of neutralizing with an alkali.
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, or to remove impurities by providing a filtration step.

The golden silk extract thus produced has a high ability to produce versican. For example, it is known that the presence of TGF-β, which is a skin transmitter, has an effect of improving wrinkles (see, for example, JP-A-2012-250968), and the expression level of versican mRNA in fibroblasts is increased by TGF-β. The present inventors have confirmed that it increases (positive control in Examples described later). The golden silk extract in this embodiment can have a high versican production ability that surpasses TGF-β. Specifically, with respect to the mRNA expression level (control: control) when 50% ethanol aqueous solution was added to normal human fibroblasts 5000 cells / well and cultured at 37 ° C. under 5% CO ₂ for 48 hours, It is preferable that the amount of mRNA expression when the silk extract is added to normal human fibroblasts as an evaporation residue to be 1% by mass and cultured is 1.5 times or more, preferably 2.0 times or more It is. Such an external preparation for skin having high versican production ability may contain the versican production promoter according to this embodiment.

Since the golden silk extract which concerns on this embodiment has such a high versican production ability, it can be set as the skin external preparation which has an anti-aging effect by adding to a skin external preparation.
As an external preparation for skin, it is applied as cosmetics, quasi drugs and the like.
In this embodiment, the golden silk extract concentration in the external preparation for skin is usually 0.00001% by mass or more, preferably 0.0001% by mass or more, more preferably 0.001% by mass or more, and usually 30% by mass. % Or less, preferably 10% by mass or less, and more preferably 5% by mass or less. By setting it as the said range, it can be set as the skin external preparation which has an anti-aging effect suitably.

The skin external preparation according to this embodiment can contain necessary components as appropriate according to the use such as cosmetics and quasi drugs.
Among these, it is preferable to use as cosmetics, especially anti-aging cosmetics.
When applied to cosmetics, it is possible to broadly blend the components normally used in cosmetics, and the dosage form and use are not limited at all. Hereinafter, the components that can be contained in the cosmetic when applied to the cosmetic will be described.

Examples of active ingredients include whitening ingredients, wrinkle-improving ingredients, anti-inflammatory ingredients, animal and plant extracts. In addition, you may mix | blend with the golden silk extract demonstrated above.
Content of the whitening component in cosmetics is 0.01-30 mass% normally, 0.1-10 mass% is preferable and 1-5 mass% is more preferable.

  The wrinkle improving component is not particularly limited as long as it is generally used in cosmetics. For example, vitamin A or a derivative thereof is retinol, retinal, retinoic acid, tretinoin, isotretinoin, retinoic acid tocopherol, retinol palmitate, retinol acetate or ursolic acid benzyl ester, ursolic acid phosphate ester, betulinic acid benzyl ester, benzylic acid A phosphate ester is mentioned. Content of the wrinkle improvement component in cosmetics is 0.01-30 mass% normally, 0.1-10 mass% is preferable and 1-5 mass% is more preferable.

The extracts derived from animals and plants are not particularly limited as long as they are generally used for pharmaceuticals, cosmetics, foods and the like. For example, akebi extract, asunaro extract, asparagus extract, avocado extract, achacha extract, almond extract, arnica extract, aloe extract, aronia extract, apricot extract, ginkgo biloba extract, Indian mushroom extract, fennel extract, udo extract, ages extract, sorghum extract , Enmezo extract, Ogon extract, Oat extract, Auren extract, Panax ginseng extract, Hypericum extract, Adrianthus extract, Orange extract, Oyster extract, Cuckoo extract, Chamomile extract, Carrot extract, Kawara mugwort extract, Licorice extract, Kiwi extract, Cucumber extract, Guava extract, Kujin extract, Gardenia extract, Kumazasa extract, Clara extract, Walnut extract, Grapefruit extract, Black rice , Chlorella extract, mulberry extract, caquette extract, gentian extract, gentian extract, Gennosho extract, tea extract, burdock extract, rice extract, fermented rice extract, rice fermented extract, rice germ oil, bilberry extract, salvia extract, bonito extract , Sasa extract, Hawthorn extract, Sansha extract, Salamander extract, Shiitake extract, Giant extract, Shikon extract, Perilla extract, Linden extract, Citrus extract, Peonies extract, Pepper extract, Ginger root extract, Birch extract, Japanese cedar extract, Stevia extract, Stevia fermentation , Kizuta extract, Hawthorn extract, Elderberry extract, Yarrow extract, Pepper extract, Sage extract, Mallow Kiss, Senkyu Extract, Sembura Extract, Sakuhakuhi Extract, Daiou Extract, Soybean Extract, Taiso Extract, Thyme Extract, Dandelion Extract, Tea Extract, Clove Extract, Chimpi Extract, Green Tea Extract, Capsicum Extract, Toki Extract, Tokinsenka 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, hammamellis extract, cypress extract, hinoki extract, loquat extract, dandelion extract, fukinotou extract , Bukuryo extract, Butcher bloom extract, Grape extract, Grape seed extract, Loofah extract, Safflower extract, Peppermint extract, Bo Daijuku extract, button extract, hop extract, pine extract, maroonnier extract, mizubasho extract, mukuroji extract, melissa extract, mozuku extract, peach extract, cornflower extract, eucalyptus extract, yukinoshita extract, yuzu extract, lily extract, juniper extract, mugwort extract, lavender extract, Preferable extracts include 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, and bitumen extract. .
Content of the plant and animal origin extract in cosmetics is 0.01-30 mass% normally, 0.1-10 mass% is preferable and 1-5 mass% is more preferable.

Examples of the anti-inflammatory component include clarinone, glabrizine, glycyrrhizic acid, glycyrrhetinic acid, pantothenyl alcohol, etc., preferably glycyrrhizic acid and salts thereof,
These are alkyl glycyrrhetic acid and its salt, and glycyrrhetic acid and its salt.
Content of the anti-inflammatory component in cosmetics is 0.01-30 mass% normally, 0.1-10 mass% is preferable and 1-5 mass% is more preferable.

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 May be mentioned trimethylolpropane triisostearate, tetra-2-ethylhexyl acid pentane erythritol, tri-2-ethylhexyl acid glycerin, trimethylolpropane triisostearate.

  Furthermore, 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, cinnagar 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.

  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.

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

  Moreover, the manufacturing method of cosmetics is not specifically limited, What is necessary is just to manufacture by using suitably with a known method according to the dosage form of the cosmetics to be applied. As a dosage form when applied as a cosmetic, any of conventionally known lotion dosage forms, emulsion dosage forms, essence dosage forms, cream dosage forms, and powder-containing dosage forms can be used.

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.
<Example 1>
A mixed solvent (pH 1.83) of 400 ml of dilute ethanol solution and 1 ml of concentrated hydrochloric acid in which 10 g of golden silk (Golden Silk No. 11 from Tonhuachachan, Lampung, Thailand) is added to 1 volume of 95% ethanol. 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 extract in which the starch was precipitated was filtered to obtain a golden silk extract 1.

<Example 2>
A golden silk extract 2 was obtained in the same manner as in Example 1 except that the extraction solvent was a mixed solvent of 400 ml of 99.5% ethanol solution and 1 ml of concentrated hydrochloric acid.

<Comparative Example 1>
A golden silk extract 3 was obtained in the same manner as in Example 1 except that the extraction solvent was extracted with a mixed solvent of 400 ml of water and 1 ml of 6M sodium hydroxide solution and then neutralized with 1M hydrochloric acid.
<Comparative Example 2>
A silk extract was obtained in the same manner as in Example 1 except that 10 g of white silk fiber produced in Japan was immersed in a mixed solvent (pH 1.83) of 400 ml of water and 1 ml of concentrated hydrochloric acid and extracted at 120 ° C. for 1 hour. 4 was obtained.

<Comparative Example 3>
The golden silk extract was used in the same manner as in Example 1 except that the extraction solvent was a mixed solvent of 400 ml of dilute ethanol solution in which 1 volume of water was added to 1 volume of 95% ethanol and 1 ml of concentrated hydrochloric acid, and the extraction temperature was 40 ° C. 5 was obtained.

<Comparative Example 4>
A golden silk extract 6 was obtained in the same manner as in Comparative Example 3 except that the extraction temperature was room temperature and the extraction time was 6 days.

<Versican production promotion evaluation>
Each of the above extracts was added to normal human fibroblasts 5000 cells / well to a concentration of 1% by mass as an evaporation residue, and cultured at 37 ° C. under 5% CO ₂ for 48 hours. After culturing, 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. In addition, as a control, mRNA expression level was measured in the same manner when a control (control) in which 50% ethanol was added instead of the extract and 2 ng of TGF-β having anti-aging action was added as a positive control. The results are shown in FIGS.

As shown in FIG. 1, when extracts 1 and 2 using acid alcohol as an extraction solvent were added to fibroblasts, the expression level of versican mRNA was 2.0 times or more that of the control (control). It can be understood that it has an excellent versican production promoting action. In particular, the extract 1 having a high alcohol concentration in the extract has an mRNA expression level about 2.5 times that of the control and has a very high versican production promoting effect. On the other hand, when extracts 3 and 4 extracted with a solvent containing no alcohol were added to fibroblasts, the mRNA expression level was about 1.2 times that of the control.
Moreover, as shown in FIG. 2, when the extract 5 extracted at 40 ° C. or the extract 6 extracted at room temperature was added to the fibroblasts, the expression level of versican mRNA was smaller than that of the control. That is, it can be seen that an extract extracted at a temperature higher than 60 ° C. has a high versican production promoting action.
As described above, the golden silk extract has a versican production-promoting action, and a golden silk extract having a high versican production-promoting action can be produced by extraction through a specific step.

  The versican production promoter of the present invention can be suitably blended in a skin external preparation, particularly an anti-aging cosmetic.

Claims (5)

  Extracting the golden silk with a solvent at 60 ° C. or higher, and a method for producing the golden silk extract.   The production method according to claim 1, wherein the solvent is an acidic alcohol solvent or an acidic mixed solvent of alcohol and water.   The versican production promoter which consists of a golden silk extract manufactured by the manufacturing method of Claim 1 or 2.   The skin external preparation containing the versican production promoter of Claim 3.   The external preparation for skin according to claim 4, which is an anti-aging cosmetic.