JP2010138153A - External preparation for skin for ultraviolet prevention - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a means for protecting the skin from receiving damage by light irradiation. <P>SOLUTION: The external preparation for the skin contains [1] a YAC compound having the following characteristics and/or a salt thereof, and [2] pantetheine-S-sulfonic acid and/or a salt thereof: (1) having λmax at 405-415 nm and 660-670 nm; (2) having a specific<SP>1</SP>H-NMR spectrum; (3) exhibiting a single peak about at 15 min in HPLC analysis under the following conditions: mobile phase: 90% acetonitrile; column: ODS 4.6×250 mm; flow velocity: 1 mL/min; temperature: 40°C; and detection: ultraviolet region of 210 nm; and (4) having a chemical composition formula of C<SB>34</SB>H<SB>40</SB>O<SB>9</SB>, and mass analysis spectrum of 593(M+H). <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an external preparation for skin, and more particularly to an external preparation for skin suitable for cosmetics.

  There are various effects of light, especially ultraviolet rays on living organisms. For example, DNA breakage, gene damage due to cleavage, generation of cancer induced by it, formation of peroxides in lipids, and formation of biological components by the peroxides. Examples include oxidative damage, increased expression of proteases typified by matrix metalloproteases such as collagenase and gelatinase, and thereby cleavage and fragmentation of matrix proteins such as collagen (see, for example, Patent Document 1). . Furthermore, there is an increase in an inflammatory system reaction involving macrophages, interleukins and the like induced by such a reaction (see, for example, Patent Document 2). After such an inflammatory reaction has taken place, there is no choice but to rely on non-steroidal anti-inflammatory agents, but in such a response, melanin deposition often occurs after inflammation subsides. These reactions are all irreversible reactions, and it can be said that it is more important to avoid such an injury than to treat them immediately after receiving them.

  As preventive measures to protect the living body from the influence of such light, combination of UV absorbing powders such as zinc oxide and titanium dioxide, UV absorbers such as benzophenone and cinnamic acid derivatives, and collagen crosslinking inhibitors consisting of various herbal extracts Are generally employed (see, for example, Patent Document 3 and Patent Document 4). However, these treatments are treatments in which light is not applied to the skin, and it can be said that there was not much to do with respect to the influence of leaking light such as makeup collapse. Furthermore, when the inflammatory system is enhanced, it cannot be denied that the current status is fully maintained.

On the other hand, advanced glycation end products (hereinafter sometimes abbreviated as AGEs) play an important role in the skin aging phenomenon. It is known that there is a leaf extract or the like (see, for example, Patent Document 5, Patent Document 6, and Patent Document 7). Among these, the present inventors have refined an active ingredient of AGEs decomposition action in a mugwort extract and have found a YAC compound having the following chemical characteristics. YAC compounds having such property values are not known in the literature.
<Characteristics of YAC compound>
(1) It has (lambda) max in 405-415 nm and 660-670 nm.
(2) 1H-NMR spectrum shown in FIG.
(3) It has a 13C-NMR spectrum shown in FIG.
(4) In HPLC analysis under the following conditions, a single peak is shown around 15 minutes.
Mobile phase: 90% acetonitrile column: ODS 4.6 × 250 mm
Flow rate: 1 ml / min.
Temperature: 40 ° C
Detection: UV part 210nm

  In addition, pantethein-S-sulfonic acid and / or a salt thereof has an excellent whitening action and cell inactivation action (see, for example, Patent Document 8, Patent Document 9, and Patent Document 10), and is a material useful as a cosmetic material. It is widely used in the cosmetics field. However, nothing is known about the preventive effect against damage to the living body caused by light, especially, ultraviolet irradiation. In addition, there is no known relationship between such ingredients and extracts of the genus Artemisia. There is also no known relationship between AGEs and pantethein-S-sulfonic acid and / or its salts.

JP 2004-217670 A Special table 2008-523083 JP 09-136824 A Japanese Patent Laid-Open No. 11-124323 JP 2001-122758 A JP 2001-108622 A JP 2007-161663 A JP 2005-139070 A JP 09-59142 A JP 09-110645 A

  The present invention has been made under such circumstances, and an object of the present invention is to provide means for protecting the skin from being damaged by light irradiation.

In view of such a situation, the present inventors have sought a means for protecting the skin from being damaged by light irradiation, and as a result of intensive research efforts, 1) a YAC compound having the following characteristics and It has been found that an external preparation for skin containing such a salt and / or a salt thereof and 2) pantethein-S-sulfonic acid and / or a salt thereof exerts such an action, thereby completing the invention.
<Characteristics of YAC compound>
(1) It has (lambda) max in 405-415 nm and 660-670 nm.
(2) 1H-NMR spectrum shown in FIG.
(3) It has a 13C-NMR spectrum shown in FIG.
(4) In HPLC analysis under the following conditions, a single peak is shown around 15 minutes.
Mobile phase: 90% acetonitrile column: ODS 4.6 × 250 mm
Flow rate: 1 ml / min.
Temperature: 40 ° C
Detection: UV part 210nm
(5) The chemical composition formula is C ₃₄ H ₄₀ O ₉ and the mass spectrometry spectrum is 593 (M + H).
That is, the present invention is as follows.
<1> A skin external preparation comprising 1) a YAC compound and / or a salt thereof having the following characteristics, and 2) pantethein-S-sulfonic acid and / or a salt thereof.
<Characteristics of YAC compound>
(1) It has (lambda) max in 405-415 nm and 660-670 nm.
(2) 1H-NMR spectrum shown in FIG.
(3) It has a 13C-NMR spectrum shown in FIG.
(4) In HPLC analysis under the following conditions, a single peak is shown around 15 minutes.
Mobile phase: 90% acetonitrile column: ODS 4.6 × 250 mm
Flow rate: 1 ml / min.
Temperature: 40 ° C
Detection: UV part 210nm
(5) The chemical composition formula is C34H40O9, and the mass spectrometry spectrum is 593 (M + H).
<2> The external preparation for skin according to <1>, wherein the YAC compound is contained as an extract of the above-ground part of Asteraceae or Asteraceae.
<3> The content of the YAC compound in the above-ground extract of the Asteraceae Artemisia or Asteraceae Artemisia is 10-6 mass% to 10-2 mass%, <2> The skin external preparation described.
<4> Further, ascorbic acid, ascorbic acid derivatives, and salts thereof, arbutin and salts thereof, and ellagic acid and one or more selected from the salts thereof, <1> to <3> The skin external preparation according to any one of <1> to <3>.
<5> The skin external preparation according to any one of <1> to <4>, further comprising grabridine, glycyrrhetic acid, glycyrrhizic acid, glycyrrhetinic acid alkyl ester, glycyrrhetinic acid and salts thereof .

  According to the present invention, it is possible to provide means for protecting the skin from being damaged by light irradiation.

<1> YAC compound which is an essential component of the external preparation for skin of the present invention The YAC compound which is an essential component of the cosmetic of the present invention has the following properties.
(Properties)
(1) It has (lambda) max in 405-415 nm and 660-670 nm.
(2) 1H-NMR spectrum shown in FIG.
(3) It has a 13C-NMR spectrum shown in FIG.
(4) In HPLC analysis under the following conditions, a single peak is shown around 15 minutes.
(5) The chemical composition formula is C ₃₄ H ₄₀ O ₉ and the mass spectrometry spectrum is 593 (M + H).
The ultraviolet / visible absorption spectrum of the YAC compound which is an essential component of the cosmetic of the present invention is shown in FIG. From this figure, it is possible to clearly discriminate the feature that λmax exists at 405 to 415 nm and 660 to 670 nm. When this compound, which is an essential component of the external preparation for skin of the present invention, is specified, such ultraviolet / visible absorption characteristics are very advantageous. That is, using HPLC equipped with a multi-wavelength detector, analysis was performed at 210 nm absorption, and the absorption at 405 to 415 nm and 660 to 670 nm was confirmed for the peak. The probability of being the YAC compound of the present invention is very high. In this sense, it is an effective confirmation means. The YAC compound of the present invention is present in the nonpolar part of the polar solvent extract. For this reason, it can be concentrated by extracting with a solvent, removing the extraction solvent by concentration under reduced pressure, etc., then separating with ethyl acetate and water, and collecting the ethyl acetate phase. This can be isolated by subjecting it to fractional purification in a chloroform / methanol mixture system using silica gel column chromatography or the like. Whether or not it is isolated can be identified by determining whether or not it is a single peak (retention time around 15 minutes) by HPLC analysis under the following conditions. FIG. 4 shows an analysis example. In this case, the retention time is 14.7 minutes.
(HPLC conditions)
Mobile phase: 90% acetonitrile column: ODS 4.6 × 250 mm
Flow rate: 1 ml / min.
Temperature: 40 ° C
Detection: UV part 210nm
(5) The chemical composition formula is C34H40O9, and the mass spectrometry spectrum is 593 (M + H).

  The YAC compound, which is an essential component of the external preparation for skin of the present invention, is a polar solvent, for example, an organic solvent that may contain water, such as methanol, ethanol, isopropyl alcohol. , Alcohols such as n-butanol, propylene glycol and 1,3-butanediol, ketones such as acetone and methyl ethyl ketone, ethers such as diethyl ether, isopropyl ether and tetrahydrofuran, halogenated hydrocarbons such as chloroform and dichloromethane, acetic acid By extracting with carboxylic acid esters such as ethyl and methyl formate, and nitriles such as acetonitrile, an extract containing the YAC compound can be obtained. As described above, liquid-liquid extraction, column chromatography, etc. It can be isolated and purified by purification means. As the extraction solvent, a hydrous alcohol is particularly preferred, and a 70 to 90% aqueous ethanol solution is particularly preferred. It is preferable to use the above-ground part for the plant part used for extraction. Prior to extraction, the plant body is preferably subjected to a fragmentation treatment such as chopping or drying and crushing. The extraction can be performed by immersing the plant or the processed product in a solvent for several days at room temperature and for several hours at a temperature near the boiling point.

  The YAC compound thus obtained exhibits a single spot in thin-layer chromatography (developing solution, chloroform: methanol = 95: 5 to 8: 2) using silica gel as a carrier, and exhibits AGE decomposition activity. The AGEs degradation activity can be quantified simply using the strength of the cleavage activity of α-diketone as an index. That is, it is incubated with 1-phenyl-1,2-propanedione, benzoic acid generated by cleavage is quantified by absorbance, and it can be determined that the greater the amount of benzoic acid produced, the higher the AGEs resolution. As an evaluation closer to vivo, there is a method in which AGEs prepared by actually incubating glucose and bovine serum albumin are decomposed, the amount of decomposition is quantified, and the AGEs resolution is quantified using the amount of decomposition as an index. YAC, which is an essential component of the cosmetic composition of the present invention, exhibits a degradation rate of about 40 to 60% in the degradation of α-diketones when the AGEs resolution is quantified by such a method, and 10% for bovine albumin AGEs. Decomposition rate of about 65 to 80% is exhibited at -3 mass%. Utilizing this property, the YAC compound, which is an essential component of the external preparation for skin of the present invention, is blended into an external preparation for skin such as cosmetics as an AGEs decomposing agent, and AGEs generated by light irradiation etc. are rapidly decomposed and accumulated. Can be prevented. This action is also related to the action of reducing the damage received on the skin by light, and is considered to be the effect of the external preparation for skin of the present invention. Therefore, in the external preparation for skin of the present invention, it is preferable to contain a YAC compound in a dose that clearly expresses the AGEs decomposition action. The procedure of these evaluation methods is shown below.

<Measurement of CC bond cleavage ability of α-diketone>
1 ml of 22 mM 1-phenyl-1,2-propanedion / MeOH + 0.1 M phosphate buffer (PH7.4) and 1 ml of measurement sample were mixed and reacted at 37 ° C. for 10 hours. The amount of benzoic acid was determined by HPLC. Quantify.
(HPLC conditions)
・ Analysis conditions Detector: Ultraviolet absorptiometer (measurement wavelength: 260 nm)
-Column: Tosoh TSK-ODS80TsQA Column temperature: Room temperature-Moving bed: Glacial acetic acid 2 g / acetonitrile 500 ml + edetate disodium solution (1 → 250) 500 ml Flow rate: 1 ml / min

<Measurement of glucose-bovine serum albumin AGE resolution>
The materials used are as follows.
AGE-BSA: Glucose and BSA are incubated at 37 ° C. for 12 weeks or more,
PD-10 columns (Amersham Biosciences 17-0851-01) from which excess glucose was removed Primary antibody: Anti-Albumin, Bovine Serum, Rabbit-Poly ROCKLAND 201-14331 / 20000
Secondary antibody: Goat anti-rabbit IG ghorseradish
peroxidase conjugate Bio RAD 170-6515 1/10000
Substrate: TMB solution Wako 546-01911
(procedure)
100 μl of 10 μg / ml AGE-BSA was added to a type I collagen-coated 96-well microplate (Bio Coat 35 4407), (1.0 μg AGE-BSA / well) was left at 37 ° C. for 4 hours, and then 0.05% Wash three times with Tween20 / PBS (−) (shake on a micromixer at room temperature for 3 minutes), add 100 μl of each concentration sample dissolved in PBS (−), and react at 37 ° C. for 10 hours or more. Then, it wash | cleans 3 times by 0.05% Tween20 / PBS (-), 100 microliters / well of primary antibodies are added to each well, and it leaves still for 30 minutes at room temperature. Wash 3 times with 0.05% Tween20 / PBS (−), add 100 μl / well of secondary antibody, and let stand at room temperature for 30 minutes. Wash 3 times with 0.05% Tween20 / PBS (−), add 100 μl / well of TMB, and react at room temperature for 15 minutes. Add 100 μl / well of 1N HCl, stop the reaction, and measure the absorbance at 450 nm. The amount of AGEs was changed, a calibration curve was drawn, and the amount of residual AGEs was quantified from this calibration curve. The AGEs decomposition rate was calculated by subtracting the remaining AGEs from the added AGEs, dividing by the added AGEs, and multiplying by 100.

  The YAC compound, which is an essential component of the external preparation for skin of the present invention, is considered to have a reactive substituent such as a hydroxyl group from the NMR data in FIGS. 1 and 2, and is led to a derivative using such a reactive group. I can do it. Such a derivative is a derivative of a YAC compound that is an essential component of the external preparation for skin of the present invention. Examples of the derivatives of the present invention include reacting alkyl ethers, alkyl esters, acylated products obtained by reacting acyl halides, monoethanolamine, etc., alkylated with a halogenated hydrocarbon such as methyl iodide. Preferred examples include amides that have been prepared. As long as they have AGEs resolution in the above evaluation method, these derivatives belong to the technical scope of the present invention as essential components of the external preparation for skin of the present invention.

<2> Pantethein-S-sulfonic acid which is an essential component of the skin external preparation of the present invention The skin external preparation of the present invention contains pantethein-S-sulfonic acid and / or a salt thereof as an essential component. Pantethein-S-sulfonic acid (hereinafter sometimes abbreviated as PSS) is a known compound represented by the following structural formula (1), which is naturally present in ginseng and promotes the growth of Bifidobacterium. It is also known to have a whitening effect, a hair-growth effect, and blood circulation promotion that are useful in cosmetics. In the present invention, PSS can be used not only as a free acid but also in a salt form. Examples of the salt include organic acid salts and inorganic acid salts, and alkali metal salts and alkaline earth metal salts are preferable. In particular, the calcium salt form is particularly preferred when blended. In the external preparation for skin of the present invention, such a component works together with the YAC compound and has an effect of reducing the damage caused by light on the skin. In order to express such effects clearly, the PSS and / or salt thereof is preferably contained in a total amount of 0.01 to 5% by mass, more preferably 0%, based on the total amount of the external preparation for skin. 0.05 to 2% by mass.

<3> External preparation for skin of the present invention The external preparation for skin of the present invention contains the essential component, and has an effect of suppressing the occurrence of light irradiation injury in skin cells. The external preparation for skin of the present invention is administered to the skin prior to light irradiation, and has an action of erasing the damage received on the skin by light irradiation at an initial stage. The skin external preparation of the present invention is applied to skin external medicines, cosmetics (including quasi-drugs), skin external goods and the like, but is particularly preferably applied to cosmetics. As cosmetics, for example, it is preferably applied to basic cosmetics such as lotions, milky lotions and creams. A particularly preferred system is an emulsifier form, which is preferably contained in the inner water phase of a water-in-oil emulsifier form. Examples of the water-in-oil emulsifier form include 0.1 to 10% by weight of an organically modified clay mineral such as distearyldimonium chloride modified hectorite, which is commercially available under the name “Benton 38V”, and polyether modified methylpolyethylene. Preferred examples include an emulsifying system in which a surfactant is combined with 0.1 to 5% by mass of siloxane.

  In the external preparation for skin of the present invention, optional components usually used in external preparations for skin can be contained in addition to the essential components. Such optional ingredients include, for example, macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, palm oil, palm oil, liquid Lanolin, hydrogenated palm oil, hydrogenated oil, molasses, hydrogenated castor oil, beeswax, candelilla wax, carnauba wax, ibotarou, lanolin, reduced lanolin, hard lanolin, jojoba wax and other oils, waxes; liquid paraffin, squalane, pristane, ozokerite, Hydrocarbons such as paraffin, ceresin, petrolatum, microcrystalline wax; higher fatty acids such as oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, undecylenic acid; cetyl alcohol, stearyl alcohol -Le, Isosteari Higher alcohols such as alcohol, behenyl alcohol, octyl decanol, myristyl alcohol, cetostearyl alcohol; cetyl isooctanoate, isopropyl myristate, hexyldecyl isostearate, diisopropyl adipate, sebacic acid Di-2-ethylhexyl, cetyl lactate, diisostearyl malate, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, glycerin di-2-heptylundecanoate, glycerin tri-2-ethylhexanoate Synthetic ester oils such as trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, pentane erythritol tetra-2-ethylhexanoate; dimethylpolysiloxane, methylphenylpoly Chain polysiloxanes such as Loxane and diphenylpolysiloxane; Cyclic polysiloxanes such as octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, and dodecamethylcyclohexanesiloxane; Amino-modified polysiloxane, polyether-modified polysiloxane, and alkyl-modified polysiloxane , Oil agents such as silicone oils such as modified polysiloxane such as fluorine-modified polysiloxane; anionic surfactants such as fatty acid soap (sodium laurate, sodium palmitate, etc.), potassium lauryl sulfate, triethanolamine alkyl sulfate ; Cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride, laurylamine oxide; imidazoline-based amphoteric surfactants (2-cocoyl-2-imida) Zolinium hydroxide-1-carboxyethyloxy disodium salt, etc.), betaine surfactants (alkyl betaine, amide betaine, sulfobetaine, etc.), and amphoteric surfactants such as acylmethyltaurine; sorbitan fatty acid esters (sorbitan) Monostearate, sorbitan sesquioleate, etc.), glycerin fatty acids (such as glyceryl monostearate), propylene glycol fatty acid esters (such as propylene glycol monostearate), hardened 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 ester Tells (POE-glycerol monoisostearate, etc.), POE fatty acid esters (polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (POE2-octyldodecyl ether, etc.), POE alkylphenyl ethers (POE nonylphenyl ether, etc.), Pluronic types, POE / POP alkyl ethers (POE / POP2-decyltetradecyl ether, etc.), Tetronics, POE castor oil, Non-ionic surfactants such as hydrogenated castor oil derivatives (POE castor oil, POE hydrogenated castor oil, etc.), sucrose fatty acid ester, alkyl glucoside; polyethylene glycol, glycerin, 1,3-butylene glycol, erythritol , Sorbitol, xylitol, maltitol, propylene Ricol, dipropylene glycol, diglycerin, isoprene glycol, 1,2-pentanediol, 2,4-hexanediol, 1,2-hexanediol, 1,2-octanediol, etc. Moisturizing ingredients such as sodium pyrrolidone carboxylate, lactic acid, sodium lactate; surface treated mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, silicic anhydride (Silica), powders such as aluminum oxide and barium sulfate; inorganic pigments such as bengara, yellow iron oxide, black iron oxide, cobalt oxide, ultramarine, bitumen, titanium oxide, zinc oxide, which may be treated on the surface PAR; agents whose surface may be treated, such as titanium mica, fish phosphorus foil, bismuth oxychloride; red 202 which may be laked, red Color 228, Red 226, Yellow 4, Blue 404, Yellow 5, Red 505, Red 230, Red 223, Orange 201, Red 213, Yellow 204, Yellow 203, Blue 1 No., green 201, purple 201, red 204, etc .; organic powders such as polyethylene powder, polymethyl methacrylate, nylon powder, organopolysiloxane elastomer; paraaminobenzoic acid UV absorbers; Anthranilic acid UV absorbers; salicylic acid UV absorbers; cinnamic acid UV absorbers; benzophenone UV absorbers; sugar UV absorbers; 2- (2′-hydroxy-5′-t-octylphenyl) UV absorbers such as benzotriazole and 4-methoxy-4'-t-butyldibenzoylmethane; lower grades such as ethanol and isopropanol Alcohols; vitamin A or derivatives thereof, vitamin B6 hydrochloride, vitamin B6 tripalmitate, vitamin B6 dioctanoate, vitamin B2 or derivatives thereof, vitamin B such as vitamin B12, vitamin B15 or derivatives thereof; a-tocopherol , S-tocopherol, γ-tocopherol, vitamin E such as vitamin E acetate, vitamin D, vitamin H, pantothenic acid, panthetin, vitamins such as pyrroloquinoline quinone, etc .; phenoxyethanol, etc. Preferred examples of the antibacterial agent include organic modified clay minerals such as hectorite and dimethyl distearyl ammonium modified hectorite. The cosmetic of the present invention can be produced by treating these essential components and optional components according to a conventional method.

  Among these optional components, particularly preferred are ascorbic acid, ascorbic acid derivatives, and salts thereof, arbutin and salts thereof, and ellagic acid and salts thereof, isoflavan, isoflavanone, kojic acid and salts thereof. Among them, ascorbic acid, ascorbic acid derivatives, and salts thereof, arbutin and salts thereof, and ellagic acid and salts thereof are particularly selected from one or more kinds. preferable. Such a component can contain only one species or a combination of two or more species. The content for providing a preferable addition effect to the above effect is 0.1 to 10% by mass, and more realistically 0.5 to 5% by mass.

  In addition, tranexamic acid, tranexamic acid alkylamides such as tranexamic acid methylamide, grabrizine, glycyrrhetic acid, glycyrrhizic acid, glycyrrhetinic acid alkyl ester, glycyrrhetinic acid, and salts thereof can also be erased. It is preferable in terms of post-treatment due to the influence of light. Particularly preferably, it contains glabrizine, glycyrrhetinic acid, glycyrrhizic acid and salts thereof. Such a component can contain only one species or a combination of two or more species. Content for exhibiting the said effect is 0.1-10 mass%, More realistically, it is 0.5-5 mass%.

 The skin external preparation of this invention can be manufactured by processing these essential components and arbitrary components according to a conventional method.

<Production Example 1>
After chopping 100 g of dry matter of the whole plant of Artemisia genus Artemisia, 500 ml of methanol is added and heated to reflux for 3 hours. After cooling, insolubles are removed by filtration, followed by concentration under reduced pressure, followed by freezing. The extract 1 was obtained by drying. Thereafter, 200 ml of water and 200 ml of ethyl acetate were added to Extract 1 to perform liquid-liquid extraction, and the ethyl acetate phase was collected and concentrated under reduced pressure to obtain Fraction 1. Fraction 1 was concentrated under reduced pressure and then purified by silica gel column chromatography. That is, the silica gel is wetted with chloroform, packed in a column, and the concentrate of the extract 2 dissolved in chloroform is charged. Chloroform, chloroform containing 1% methanol, chloroform containing 5% methanol, chloroform containing 10% methanol and then 15% 50 ml of methanol-containing chloroform was allowed to flow, and the effluent was concentrated under reduced pressure. These fractions were sequentially designated as Fraction 2, Fraction 3, Fraction 4, Fraction 5, and Fraction 6. When the cleavage activity of the a-diketone was examined for fractions 1 to 6, fraction 3 was the highest and was 48%. Fraction 3 was further purified four times by silica gel column chromatography using a chloroform / methanol mixture as an elution solvent (first time chloroform: methanol = 100: 0 → 1: 1, second time chloroform: methanol = 100: 2 → 9). 1: 3rd chloroform: methanol = 100: 5 → 8: 2, 4th chloroform: methanol = 100: 0 → 100: 5), silica gel thin layer chromatography (developing solvent chloroform: methanol = 95: 5), Amorphous fraction 7 as a single spot was obtained by coloration by baking with 50% sulfuric acid aqueous solution. The NMR of this product is shown in FIGS. A chart on HPLC (conditions are as follows) is shown in FIG. The ultraviolet absorption spectrum is shown in FIG. From these results, although the structure itself is unknown, it is presumed that the fraction 7 is a single substance.

<Measurement of glucose-bovine serum albumin AGE resolution of fraction 7>
The AGEs degradation action at various concentrations of Fraction 7 against AGEs previously produced from glucose and bovine serum albumin by the above procedure was measured. The results are shown as a graph in FIG. From this, it can be seen that the presence of 10-5% by mass of the compound of the present invention in fraction 7 exhibits the effectiveness of its AGE decomposition. Further, when the compound of fraction 7 is contained as an extract of a plant body of a plant such as mugwort or a purified product thereof, whether or not the AGEs are decomposed at the blending concentration in the extract or the purified product. When the AGEs decomposition action is confirmed, a simple discrimination is made so that the extract or the purified product can be contained in a skin external preparation such as cosmetics. It is also possible to determine the inclusion in an external preparation for skin.

<Effect on light>
A 96-well plate was inoculated with 5 × 10 4 cells per well of normal human epidermis melanocytes (Kurabo) and cultured in Medium 154S medium (Kurabo) for 24 hours under conditions of 37 ° C. and 5% CO 2. Defined keratinocyte to which no supplement was added -SFM medium (Gibco BRL) after medium exchange, 1 μg / mL (final concentration) of pantethein-S-sulfonic acid in the presence of fraction 7 of 1 μg / mL (final concentration) Cultivation was continued for 3 days in the presence of fraction 1 at 1 μg / mL (final concentration) and pantethein-S-sodium sulfonate at 1 μg / mL (final concentration) in the presence of sodium. After exchanging with 100 μl of SFM medium (Gibco BRL), ultraviolet light 10 μJ was irradiated, and apoptotic cells were quantified using an LDH detection kit commercially available from TaKaRa. The positive control was irradiated with ultraviolet rays and contained no sample, and the negative control was non-ultraviolet irradiated and contained no sample. The apoptosis suppression rate of the specimen was calculated by setting the positive control apoptosis suppression rate to 0% and the negative control apoptosis suppression rate to 0%. The results are shown in Table 1. This shows that the combination suppresses apoptosis. That is, when combined with the above test, it is speculated that the combination of fraction 7 and pantethein-S-sodium sulfonate prevents MMP that is enhanced by light irradiation from damaging cells and inducing apoptosis. Is done.

  According to the formulation shown below, cosmetic 1 (water-in-oil emulsifier type), which is an external preparation for skin of the present invention, was produced. That is, each of the ingredients (a) and (b) was heated to 75 ° C., and (b) was gradually added and emulsified with stirring to homogenize the emulsified particles, followed by stirring and cooling to obtain Cosmetics 1. In the same manner, Comparative Example 1 in which fraction 7 was replaced with water, Comparative Example 2 in which pantethein-S-sodium sulfonate was replaced with water, and fraction 7 and pantethein-S-sodium sulfonate were replaced with water Comparative Example 3 was also prepared in the same manner.

<Test Example 1>
About the cosmetics 1 and Comparative Examples 1-3, the effect | action with respect to acute photodamage was investigated. That is, five 2 cm × 4 cm sites were prepared on the forearm of a paneler with a known MED (minimum erythema volume), and 40 μL of four types of specimens and water were administered to each site. 30 minutes after administration, the specimen was wiped off with absorbent cotton moistened with water, 1 hour later, irradiated with UV light twice that of MED, and 24 hours after irradiation, the color difference between the site and the non-irradiated site was measured with the Konica Minolta CR400 color difference meter. Measured color. The results are shown in Table 4 as erythema inhibition rate. From this, it can be seen that acute light damage can also be suppressed by pretreatment of the cosmetic 1 which is an external preparation for skin of the present invention. The erythema suppression rate was calculated by (100− (color difference of specimen site) / (color difference of water administration site) × 100).

  Similarly to cosmetic 1, cosmetics 2 to 4 which are external preparations for skin of the present invention were produced according to the formulation shown below, and the erythema suppression rate was determined by the same procedure as in Test Example 1. The results are shown in Table 5.

  Similarly to cosmetic 1, cosmetics 5 to 9, which are external preparations for skin of the present invention, were produced according to the formulation shown below, and the erythema suppression rate was determined by the same procedure as in Test Example 1. The results are shown in Table 7.

  The present invention can be applied to external preparations for skin such as cosmetics.

It is a figure which shows NMR of the compound of the fraction 7. It is a figure which shows NMR of the compound of the fraction 7. It is a figure which shows the HPLC chart of the compound of the fraction 7. It is a figure which shows the ultraviolet absorption spectrum of the compound of the fraction 7. It is a figure which shows the glucose-bovine serum albumin AGE resolution | decomposability of the fraction 7 in manufacture example 1. FIG. It is a figure which shows the mass spectrometry spectrum of the compound of the fraction 7.

Claims (5)

A skin external preparation comprising 1) a YAC compound and / or a salt thereof having the following characteristics, and 2) pantethein-S-sulfonic acid and / or a salt thereof.
<Characteristics of YAC compound>
(1) It has (lambda) max in 405-415 nm and 660-670 nm.
(2) 1H-NMR spectrum shown in FIG.
(3) It has a 13C-NMR spectrum shown in FIG.
(4) In HPLC analysis under the following conditions, a single peak is shown around 15 minutes.
Mobile phase: 90% acetonitrile column: ODS 4.6 × 250 mm
Flow rate: 1 ml / min.
Temperature: 40 ° C
Detection: UV part 210nm
(5) The chemical composition formula is C ₃₄ H ₄₀ O ₉ and the mass spectrometry spectrum is 593 (M + H). The external preparation for skin according to claim 1, wherein the YAC compound is contained as an extract of the above-ground part of Asteraceae or Asteraceae. The skin according to claim 2, wherein the content of the YAC compound in the above-ground extract of the Asteraceae or Asteraceae is 10-6 mass% to 10-2 mass%. Topical agent. Furthermore, ascorbic acid, ascorbic acid derivatives, and salts thereof, arbutin and salts thereof, and one or more selected from ellagic acid and salts thereof, The skin external preparation of any one of claim | item 1-3. Furthermore, the skin external preparation in any one of Claims 1-4 characterized by containing a glabrizine, a glycyrrhetic acid, a glycyrrhetic acid alkyl ester, a glycyrrhizic acid, and these salts.

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