JP2007186442A - Skin care preparation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new skin care preparation, excellent in safety and suitable for inhibiting the melanin production. <P>SOLUTION: This skin care preparation suitable for inhibiting the melanin production is provided by containing amorisin expressed by the chemical formula of the figure and/or its salt, or an extract of a plant body containing the same so as to exhibit excellent effects for the prevention and improvement of skin pigmentation based on the melanin production-inhibiting activity. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an external preparation for skin, and more particularly to an external preparation for skin suitable for whitening cosmetics (including quasi-drugs).

  It is well known that ultraviolet rays have various adverse effects on the skin, such as causing inflammation on the skin and generating water bubbles when exposed to a large amount. In addition, even cumulative exposure causes blemishes and freckles through increased melanin production, which is also a cosmetic problem. Skin pigmentation after blemishes, buckwheat and sunburn is one of the skin problems of middle-aged and elderly people, because melanin production is significantly enhanced by the activation of pigment cells (melanocytes) present in the skin. . For the purpose of preventing and improving these skin pigment troubles, external preparations for skin containing whitening agents such as ascorbic acids, hydrogen peroxide, glutathione, colloidal sulfur, hydroquinone and catechol are known. However, ascorbic acids are easily oxidized and unstable in a water-rich system such as a water-containing cosmetic and cause discoloration. In addition, hydrogen peroxide solution has problems in storage stability and safety, and glutathione and colloidal sulfur give a remarkable off-flavor and are restricted from being used in products. Furthermore, hydroquinone, catechol, and the like have problems in safety such as skin irritation and allergic properties, and a whitening agent that is not yet fully satisfactory has not yet been obtained. In addition, it has become clear that there are various mechanisms of action regarding the skin pigmentation phenomenon, and it is becoming clear that there are compounds that act on each. Even in this sense, it has been desired to develop a novel compound that suppresses melanin production.

  It has been reported that Amorisin is contained in plants of the leguminous weasel genus Amorpha fruticosa Linn. (See, for example, Non-Patent Document 1 and Non-Patent Document 2). However, it is not known that this amorisin is contained in the plant body of the leguminous genus Kihagi (Lespedeza buergeri Miq.). On the other hand, a technique is known in which 0.1 to 10% of a certain type of flavanone compound is blended to obtain a cosmetic having a whitening action and an anti-inflammatory action (see, for example, Patent Document 1). However, amoricin has a strong melanin production inhibitory action, and it has not been known regarding a skin external preparation for inhibiting melanin production containing 0.0001 to 5% by mass of this amorisin.

  On the other hand, regarding the plant extract of the genus Leguminosae, it is known that the extract of the genus Legumidae (Lespedeza bicolor Turcz.) Has an anti-androgenic action (see, for example, Patent Document 2). Furthermore, it is known that various isoflavones are contained in the leguminous genus Marvahagi (Lespedeza cyrtobotrya Miq.) (See, for example, Non-Patent Documents 3 and 4), and isoflavenes. It is also known that Haginin A, Haginin B, Haginin C, Haginin D, etc., which are one of the above, have a melanin production inhibitory action (see, for example, Patent Document 3). Furthermore, it is known also about the cosmetics containing the isoflavenes derived from the leguminous genus Marubahagi (for example, refer patent document 4). However, the extract of leguminous genus Kihagi and its polar solvent extract have a melanin production inhibitory effect, and further, the production of melanin containing the extract of the leguminous genus Kihagi plant body. There is no known skin external preparation suitable for suppression.

Japanese Patent Laid-Open No. 06-16531 Japanese Patent Laid-Open No. 06-128171 JP-A-11-322530 International Publication Pamphlet No. 03/097004 Heterocycles, Vol. 19, No. 10, 1793-1796, (1982) Studies in Organic Chemistry, Vol.11, 245-250, (1982) Chem. Pharm. Bull., Vol. 28, No. 4, 1172-1177, (1980) Chem. Pharm. Bull., Vol. 29, No. 8, 2205-2209, (1981)

  This invention makes it a subject to provide the skin external preparation suitable for novel melanin production suppression.

  As a result of intensive studies to solve the above problems, the present inventors have found that amorosine and / or a salt thereof, which is a kind of flavone, has a strong inhibitory effect on melanin production in pigment cells, Furthermore, it was found that when this was incorporated into a skin external preparation base, it exhibited an excellent effect of preventing and improving skin pigmentation, and the present invention was completed based on this. That is, the present invention is as follows.

(1) An external preparation for skin containing amoricin and / or a salt thereof represented by the following chemical formula.

アモリシン

Amoricin

(2) The external preparation for skin according to (1), which is used for suppressing melanin production.
(3) The external preparation for skin according to (1) or (2), wherein Amoricin is contained in an amount of 0.0001 to 5% by mass of the total external preparation for skin.
(4) A topical skin preparation characterized by containing an extract of a plant belonging to the genus Leguminosae that contains amoricin and / or a salt thereof, using a polar solvent.
(5) The external preparation for skin according to (4), wherein the extract with the polar solvent is an extract with ethanol, hydrous ethanol, polyhydric alcohol, or hydrous polyhydric alcohol.
(6) The external preparation for skin according to claim 4 or 5, wherein the extract with the polar solvent contains 0.001 to 0.1% by mass of amoricin.
(7) The skin external preparation according to any one of (1) to (6), which is a cosmetic.
(8) The external preparation for skin according to any one of (1) to (7), which is a quasi-drug.
(9) The skin external preparation according to any one of (1) to (8), further comprising one or more selected from glycyrrhizic acid and salts thereof, and alkyl glycyrrhetinate.
(10) The content of one or more selected from the above-mentioned glycyrrhizic acid and salts thereof and alkyl glycyrrhetinate is 0.05 to 0.5% by mass with respect to the entire external preparation for skin. The external preparation for skin according to (9).
(11) The external preparation for skin according to (9) or (10), wherein the preparation is for melanin production suppression and anti-inflammatory.

  ADVANTAGE OF THE INVENTION According to this invention, the skin external preparation suitable for melanin production suppression containing a novel active ingredient can be provided.

(1) Amorisin and / or salt thereof, which is an essential component of the external preparation for skin of the present invention, The external preparation for skin of the present invention contains amorisin and / or a salt thereof represented by the above chemical formula as an active ingredient. It is a feature. Amorisin was known to be contained in the root bark of the leguminous weasel weasel butterfly, but the present inventors found that it is also contained in the aerial part of the plant of the leguminous weeping genus kihagi. It was done. However, it has not been known that amorisin has a melanin production inhibitory effect and is useful for use in a skin external preparation for inhibiting melanin production.

  Amoricin obtained by any method can be used for the external preparation for skin of the present invention. For example, amoricin can be extracted from a plant containing amoricin such as leguminous genus kihagi, leguminous weasel genus weasel. When extracting from these plants, it is most preferable to use an extract extracted from a plant of the leguminous genus Kihagi. This is because the leguminous genus Kihagi has a high content in the plant body.

  When amoricin is obtained by extraction from a plant containing amoricin, it can be extracted by a normal extraction method and further purified using a hydrophobic resin column or a silica gel column. For example, amorisin is extracted from a plant using a polar solvent or the like, and the extract is further liquid-liquid extracted with an organic solvent immiscible with water and water to form an organic solvent layer immiscible with water. It can be obtained by concentrating as necessary and performing purification using a hydrophobic resin column or silica gel column. More specifically, for example, 1 part by weight of a ground portion of a leguminous genus Kihagi, alcohols such as methanol, ethanol and isopropanol; ketones such as acetone and methyl ethyl ketone; acetonitrile, propionitrile and the like Nitriles; ethers such as diethyl ether, tetrahydrofuran and dioxane; esters such as ethyl acetate and methyl acetate; polar solvents such as halogenated hydrocarbons such as chloroform and dichloromethane; If it is near the boiling point for 1 day to 1 week, it is immersed for several hours, and after removing insolubles by filtration, the solvent is distilled off by concentration under reduced pressure, etc., and this is mixed with an organic solvent (such as diethyl ether or ethyl acetate) Liquid-liquid extraction with water (immiscible organic solvent) and water, which is immiscible with water The solvent was distilled off, fractionated by silica gel column chromatography (elution solvent; chloroform / methanol = 100/0 → 98/2), etc., and further reverse phase column chromatography such as ODS column (elution solvent; 75% acetonitrile). ) And the like, and a solvent is distilled off to obtain amorisin. The amorisin thus purified can be used as an active ingredient for inhibiting melanin production.

  The quantification of the content of amorisin is performed by, for example, an absolute calibration curve method using HPLC (for example, ODS column, elution solvent: 75% acetonitrile aqueous solution, detection: UV 280 nm) using a compound preparation obtained by purification. be able to.

  In addition, an extract of a plant body such as a leguminous genus Kihagi can be used as an amoricin-containing extract as it is in the external preparation for skin of the present invention. It is also possible to use it for the skin external preparation of this invention as an extract of the plant body to contain. According to the study by the present inventors, the content of amoricin in the plant of the leguminous genus Kihagi is 50 mg to 70 mg in 1 kg of the plant. Therefore, ethanol, hydrous ethanol, 1,3-butanediol, hydrous 1,3-butanediol, propylene glycol, hydrous propylene glycol, glycerin, hydrous glycerin and the like can be used as it is added to the skin external preparation. When extracted using a polar solvent, it can be used as it is as an amorillin-containing extract in a skin external preparation. For example, when 1 kg of Kihagi plant is extracted with 1 L of polar solvent, the content of Amorisin in the extract is 0.005 to 0.007 wt / v%, and 1 kg of Kihagi plant is 5 L of polar solvent. In the case of extraction with a, the content of amoricin in the extract is 0.001 to 0.0014 wt / v%.

  Furthermore, the concentrate of the plant extract containing amorisin is redispersed in water and passed through a hydrophobic resin column such as Diaion HP-20 (manufactured by Mitsubishi Chemical), and eluted while lowering the polarity of the elution solvent. By performing such purification, the content of amorisin can be increased, and by performing such purification, an extract having a higher melanin production inhibitory effect can be obtained, and this is used as an external preparation of the present invention. It can also be contained in it, and is preferable.

  In the external preparation for skin of the present invention, amorisin can be used as it is, or it can be used in the form of a salt by treating with an alkali. The salt of amoricin is not particularly limited as long as it is a physiologically acceptable salt. Examples of physiologically acceptable salts include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium and magnesium; ammonium salts; organic ammonium salts such as triethylamine and triethanolamine; Preferred examples include basic amino acid salts such as arginine.

  The external preparation for skin of the present invention can be used in combination with one or more of amorisin and / or its salt obtained as described above, and as an extract of a plant containing amorisin. This can also be used.

(2) Skin external preparation of this invention The skin external preparation of this invention has a melanin production inhibitory effect, and it is preferable to apply to the skin external preparation for melanin production suppression or whitening. Specifically, the skin external preparation of the present invention is suitable as a skin external preparation for preventing and improving pigmentation diseases such as stains, buckwheat, dark black, and senile pigment spots caused by ultraviolet rays.

  The external preparation for skin of the present invention contains amoricin and / or a salt thereof. In the external preparation for skin of the present invention, amorisin and / or a salt thereof is obtained from a polar solvent extract of a plant containing amoricin, a solvent-removed product of the same-polar solvent extract, a crudely purified product by a column, or the like. It may be contained as a purified product containing amorisin by a silica gel column or the like. It is more preferable that such a purified product of amorisin is contained in the external preparation for skin of the present invention in that the degree of freedom of formulation is increased.

  It is preferable that the external preparation for skin of the present invention contains 0.0001 to 5 mass%, preferably 0.0002 to 1 mass%, of amorisin and / or a salt thereof with respect to the entire external preparation for skin. Is more preferable, 0.0004-0.1 mass% is still more preferable, 0.001-0.05 mass% is especially preferable. When used in skin preparations such as cosmetics for the purpose of preventing the occurrence of spots, freckles, dark black, etc. caused by sunburn (ultraviolet rays), 0.0001% by mass or more is intended to improve pigmentation When used as an external preparation for the skin, 0.001% by mass or more is preferably used as an effective amount. When the content is less than 0.0001% by mass, the melanin production-suppressing action is considerably reduced. On the other hand, even if an amount exceeding 5% by weight is used, the effect reaches a peak.

  In the external preparation for skin of the present invention, in addition to the above essential components, optional components that are usually used in cosmetics and external preparations for skin can be contained. Such optional ingredients include, for example, oils (macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil, liquid lanolin, hardened Coconut oil, hydrogenated oil, owl, hydrogenated castor oil, beeswax, candelilla wax, carnauba wax, botarou, lanolin, reduced lanolin, hard lanolin, jojoba wax, waxes, hydrocarbons (liquid paraffin, squalane, pristane, ozokerite, Paraffin, ceresin, petrolatum, microcrystalline wax, etc.), higher fatty acids (oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, undecylenic acid, etc.), higher alcohols (cetyl alcohol, stearyl, etc.) Lucol, isostearyl alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, cetostearyl alcohol), synthetic ester oils (cetyl stearate, cetyl isooctanoate, isopropyl myristate, hexyldecyl isostearate, diisopropyl adipate, diisopropyl sebacate) 2-ethylhexyl, cetyl lactate, diisostearyl malate, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, glycerin di-2-heptylundecanoate, glycerin tri-2-ethylhexanoate, tri- 2-ethylhexanoic acid trimethylolpropane, triisostearic acid trimethylolpropane, tetra-2-ethylhexanoic acid pentane erythritol, etc.), chain polysilos Sun (dimethylpolysiloxane, methylphenylpolysiloxane, diphenylpolysiloxane, dimethicone, etc.), cyclic polysiloxane (octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, dodecamethylcyclohexanesiloxane, etc.), modified polysiloxane (amino-modified polysiloxane) , Polyether-modified polysiloxane, alkyl-modified polysiloxane, fluorine-modified polysiloxane, etc.) oils such as silicone oil; fatty acid soaps such as sodium laurate and sodium palmitate; potassium lauryl sulfate, alkylethanol triethanolamine ether, etc. Anionic surfactants of: Cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride, laurylamine oxide; 2-coco Ir-2-imidazolinium hydroxide-1-carboxyethyloxyl disodium salt and other imidazoline-based amphoteric surfactants; alkyl betaine, amide betaine, sulfobetaine and other betaine surfactants; acylmethyl taurine and other amphoteric interfaces Activators: sorbitan fatty acid esters (such as sorbitan monostearate, sorbitan sesquioleate, sorbitan sesquistearate), glycerin fatty acids (such as glyceryl monostearate), propylene glycol fatty acid esters (such as propylene glycol monostearate) Glycerin alkyl ethers, POE sorbitan fatty acid esters (POE sorbitan monooleate, polysoxyethylene sorbitan monostearate, etc.), POE sorbit fatty acid esters (POE- Rubbit monolaurate, etc.), POE glycerol fatty acid esters (POE-glycerol monoisostearate, etc.), POE fatty acid esters (polyethylene glycol monooleate, POE stearic acid, POE distearic acid, etc.), POE alkyl ethers (POE). -Octyldodecyl ether, etc.), POE alkyl phenyl ethers (POE nonyl phenyl ether, etc.), Pluronic types, POE / POP alkyl ethers (POE / POP-decyl tetradecyl ether, etc.), Tetronics, POE castor oil, Nonionic 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-butanediol, Thritol, sorbitol, xylitol, maltitol, propylene glycol, 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; guar gum, quince seed, carrageenan, galactan, gum arabic, pectin, mannan, starch, xanthan gum, curdlan, methylcellulose, hydroxyethylcellulose, carboxy Methyl cellulose, methyl hydroxypropyl cellulose, chondroitin sulfate, dermatan sulfate, glycogen, heparan sulfate, hyaluronic acid, sodium hyaluronate, Lagant gum, keratan sulfate, chondroitin, mucoitin sulfate, hydroxyethyl guar gum, carboxymethyl guar gum, dextran, kerato sulfate, locust bean gum, succinoglucan, carolinic acid, chitin, chitosan, carboxymethyl chitin, agar, polyvinyl alcohol, polyvinyl pyrrolidone, Thickeners such as carboxyvinyl polymer, sodium polyacrylate, polyethylene glycol, bentonite; surface may be treated, mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, anhydrous silica (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, and zinc oxide that may be treated on the surface; surface Pearl agents such as titanium mica, fish phosphorus foil, bismuth oxychloride which may be treated; red 202, red 228, red 226, yellow 4, blue 404, which may be raked, Organics such as yellow 5, red 505, red 230, red 223, orange 201, red 213, yellow 204, yellow 203, blue 1, green 201, purple 201, red 204 Pigments; Organic powders such as polyethylene powder, polymethyl methacrylate, nylon powder, organopolysiloxane elastomers; Preservatives such as methylparaben; Buffers and pH regulators such as sodium hydrogenphosphate; Paraaminobenzoic acid UV absorption Agent, anthranilic acid UV absorber, salicylic acid UV absorber, cinnamic acid UV absorber, benzophenone UV absorber, sugar UV UV absorbers such as absorbers, 2- (2′-hydroxy-5′-t-octylphenyl) benzotriazole, 4-methoxy-4′-t-butyldibenzoylmethane; lower alcohols such as ethanol and isopropanol Vitamin A or a derivative thereof, vitamin B (vitamin B6 hydrochloride, vitamin B6 tripalmitate, vitamin B6 dioctanoate, vitamin B2 or a derivative thereof, vitamin B12, vitamin B15 or a derivative thereof), vitamin E (α-tocopherol) , Β-tocopherol, γ-tocopherol, vitamin E acetate, etc.), vitamin Ds, vitamins such as vitamin H, pantothenic acid, panthetin, pyrroloquinoline quinone, and the like.

  The skin external preparation of the present invention can contain one or more of such optional components.

  In addition to the above components, the external preparation for skin of the present invention can contain one or more selected from glycyrrhizic acid having anti-inflammatory action and / or their salts and alkyl glycyrrhetinate. It is preferable to contain such a component. The present invention is effective against pigmentation caused by ultraviolet rays and the like, but when an external preparation for skin is used for such applications, there is a high possibility of causing inflammation due to ultraviolet rays. Inflammatory reactions and the various skin reactions that accompany it enhance the melanogenesis reaction. Therefore, an inflammatory reaction is suppressed by containing the component which has such an anti-inflammatory action, As a result, the melanin production inhibitory effect of the skin external preparation of this invention improves. Moreover, by containing such a component having an anti-inflammatory action, inflammation is sedated or further inflammation is suppressed, and an increase in the amount of transdermal water transpiration is suppressed. That is, the appearance of rough skin after inflammation is suppressed.

  Such components are known as active ingredients of quasi drugs, and examples of glycyrrhizic acid and / or its salts include glycyrrhizic acid, dipotassium glycyrrhizinate, ammonium glycyrrhizinate and the like. Among them, dipotassium glycyrrhizinate is preferable, and examples of the alkyl glycyrrhetinate include stearyl glycyrrhetinate and lauryl glycyrrhetinate, among which stearyl glycyrrhetinate is preferable. The preferable content of such components is 0.05 to 0.5% by mass with respect to the total amount of the external preparation for skin, and the more preferable content is 0.05 to 0.2% by mass, and further preferable content The amount is 0.05 to 0.1% by mass.

  By containing these together with amorisin, when the skin external preparation of the present invention is administered when the skin is inflamed, such as immediately after sunburn, the inflammation is suppressed more quickly, and the skin barrier function is restored. As a result, the preventive effect of worsening the skin condition is increased, and as a result, the melanin production inhibitory effect of the external preparation for skin of the present invention is also improved. That is, the external preparation for skin of such a form is preferable as an external preparation for preventing or improving pigmentation caused by ultraviolet rays even when the skin is inflamed.

  Further, as such a skin external preparation, antibacterial polyhydric alcohol such as isoprene glycol, 1,3-butanediol, 1,2-pentanediol, 1,2-hexanediol, 1,2-octanediol, A form containing 0.5 to 20% by mass with respect to the total amount of the external preparation is also preferable. By taking such a form, it is because the compounding quantity of the component which may induce irritation | stimulation at the time of inflammation, such as paraben, can be reduced or made non-combination. A form containing these antibacterial polyhydric alcohols and substantially free of parabens is particularly preferred.

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

  In addition, as a kind of skin external preparation containing the amoricin of this invention, it is preferable to apply to the cosmetics and quasi-drugs generally used widely. The external preparation for skin of the present invention is selected from ascorbic acid phosphates, ascorbic acid glucoside, arbutin, kojic acid and other whitening agents, glycyrrhizic acid and / or salts thereof, and anti-inflammatory agents such as alkyl glycyrrhetinate The active ingredient of 1 type, or 2 or more types of quasi drugs can be contained, and it can also be set as a quasi drug, and such a form is preferable. Of course, amoricin and / or a salt thereof can be used as an active ingredient of a quasi drug. When taking such a quasi-drug form, it is preferable to display the fact that it is a quasi-drug, the efficacy as a quasi-drug, and the like from the point of view of clear usage. For example, regarding the efficacy, it is possible to display a melanin production inhibitory effect and further an anti-inflammatory effect. For example, regarding the usage mode, take an appropriate amount and include it in cut cotton etc. at a site where you want to prevent spots or pigmentation, or a site where you want to prevent pigmentation, or a site with further mild inflammation. It is possible to display that it is used after being applied, or that the use is immediately stopped when a feeling of tingling or burning is felt by the above operation.

  The external preparation for skin of the present invention can be used in any form as long as it can be applied to the skin. However, since the active ingredient penetrates the skin and exhibits the effect, it is familiar to the skin. A dosage form such as a milky lotion, cream, essence, lotion, pack or the like is more preferable.

  EXAMPLES Hereinafter, although an Example is given and description is added in detail about this invention, it cannot be overemphasized that this invention is not limited only to such an Example.

  A 1 kg of tree trunk of a leguminous genus kihagi was crushed to form chips, which were immersed in 10 L of methanol, heated to reflux for 3 hours, and then the chips were removed to obtain a methanol extract. The methanol extract was concentrated, and diethyl ether and water were added to carry out liquid-liquid extraction. Then, the diethyl ether layer was taken and concentrated to obtain a crude product (6.36 g). This was fractionated by silica gel column chromatography (eluted with chloroform / methanol = 98/2). After concentration, fractionation was performed by reversed-phase column chromatography (ODS; eluted with 75% acetonitrile) to obtain 62.4 mg of amoricin.

1H-NMR of Amorisin (in Acetone-d ₆ )
1.71 (6H, s), 1.73 (3H, s), 1.76 (6H, s), 1.89 (3H, s), 2.75 (1H, dd), 2.98 (1H, dd), 3.30-3.40 (6H, m) , 5.17-5.48 (4H, m), 6.71 (1H, s), 6.85 (1H, s)

  The following test evaluated the effectiveness of amoricin for inhibiting melanin production on pigment cells.

<Test Example 1>Amoricin's melanin production inhibitory effect test Amorphin's melanin production inhibitory action was evaluated using thiouracil ( ^14C- labeled thiouracil used in the test) specifically incorporated into cells during the melanin synthesis process. Using a 24-well plate, add 2 ml of the complete medium for melanocyte culture (manufactured by Kurashiki Boseki Co., Ltd.) to 12 wells, and further normal human melanocytes at a concentration of 1.5 × 10 ⁴ cells / cm ^2. (Kurashiki Boseki Co., Ltd.) was seeded and cultured at 37 ° C. for 24 hours in a 5% carbon dioxide atmosphere.

Thereafter, all the media were changed under the following conditions. That is, 3 wells were prepared with a new complete medium for melanocyte culture (control), and 9 wells were prepared with a complete medium for melanocyte culture (each concentration; n = 3) containing amoricin at 2.5, 5.0, and 7.5 μM concentrations. Exchanged. Furthermore, ¹⁴ C-thiouracil ( ¹⁴ C-labeled thiouracil) was added to 0.25 μCi (microcurie) to the 12 wells used. The culture was further continued for 3 days under the same conditions as described above.

After completion of the culture, the culture solution is removed from each well, washed with PBS (phosphate buffered saline), and the cells are detached from the bottom of the well using a medium containing trypsin and EDTA to obtain a cell suspension. The cells were collected by centrifugation. The number of cells was counted using a hemocytometer. Thereafter, the amount of ¹⁴ C-thiouracil in the collected cells in each well was measured with a liquid scintillation counter. The percentage of the radiation dose in the cells to which amorisin was added relative to the control radiation dose was determined and used as the melanin amount (%). Note that melanin production is suppressed when the radioactivity incorporated into each cell is less. The results are shown in Table 1.

  From the results in Table 1, amoricin shows a melanin production inhibitory action at 2.5 μM, which corresponds to 0.000123 wt / v% when calculated from the molecular weight, and shows a melanin production inhibitory action even at about 0.0001% by mass. You can see that

  1 kg of leguminous genus kihagi tree trunk was crushed into chips, which were soaked in 2 L of 50% aqueous solution of 1,3-butanediol for 1 week, then the chips were removed and 50% -1,3-butane was removed. A diol water-extract was obtained. Using the compound purified in Example 1 as a standard, this extract was analyzed by HPLC (ODS column; UV 280 nm, elution with 75% acetonitrile) and found to contain 0.0032% amoricin.

<Test Example 2>
The same melanin production suppression test as in Test Example 1 was performed using the extract obtained in Example 2. Specifically, in the medium exchange after the preculture of melanocytes, the extract prepared in Example 2 was replaced with a medium containing 4 wt / v%, 8 wt / v%, and 12 wt / v%, and cultured. The results are shown in Table 2.

  From the results shown in Table 2, an extract obtained by extracting 1 kg of a plant of kihagi with 2 L of 50% -1,3 butanediol aqueous solution at 4 wt / v% showed an inhibitory action on melanin production. At this time, the content of amoricin is 0.00012 wt / v%.

  From the results of Tables 1 and 2, amoricin used as an active ingredient for suppressing melanin production of the external preparation for skin of the present invention showed an excellent inhibitory action on melanin production. At this time, no toxicity to pigment cells was observed.

  According to the prescription shown below, the emulsion which is a skin external preparation of this invention was produced. That is, the components (A) were mixed and heated to 80 ° C. On the other hand, each component of (B) was heated to 80 degreeC. The mixture of (B) was added to the mixture of (A) and stirred to emulsify. Further, (C) was added to neutralize, and then the mixture was stirred and cooled to 35 ° C. to obtain the emulsion of Example 3. In addition, in the prescription of Example 3, what substituted amoricin for water was produced, and it was set as the emulsion of Comparative Example 1.

(A)
Behenyl alcohol 0.5% by mass
Cetyl isooctanoate 2.0 mass%
Squalane 8.0 mass%
Dimethicone 2.0 mass%
Sorbitan sesquistearate 1.5% by mass
POE (45) stearic acid 1.0 mass%
Cetyl stearate 0.5 mass%
Behenic acid 0.5% by mass
(B)
1,3-butanediol 5.0% by mass
Glycerin 5.0% by mass
1,2-octanediol 1.0% by mass
Pure water 50.0% by mass
Amorisine 0.1% by mass
Dipotassium glycyrrhizinate 0.1% by mass
(C)
Pure water 22.2% by mass
Potassium hydroxide 0.6 mass%

  According to the prescription shown below, the emulsion which is a skin external preparation of this invention was produced. That is, the components (A) were mixed and heated to 80 ° C. On the other hand, each component of (B) was heated to 80 degreeC. The mixture of (B) was added to the mixture of (A) and stirred to emulsify, and further (C) was added to neutralize, and then the mixture was stirred and cooled to 35 ° C. to obtain the emulsion of Example 4.

(A)
Behenyl alcohol 0.5% by mass
Cetyl isooctanoate 2.0 mass%
Squalane 8.0 mass%
Dimethicone 2.0 mass%
Sorbitan sesquistearate 1.5% by mass
POE (45) stearic acid 1.0 mass%
Cetyl stearate 0.5 mass%
Behenic acid 0.5% by mass
(B)
1,3-butanediol 5.0% by mass
Glycerin 5.0% by mass
1,2-octanediol 1.0% by mass
Pure water 50.0% by mass
Extract of Example 2 10.0% by mass
(As an amoricin, 0.00012 mass% contained in the emulsion)
Dipotassium glycyrrhizinate 0.1% by mass
(C)
Pure water 14.3% by mass
Potassium hydroxide 0.6 mass%

<Test Example 2> Efficacy test of the external preparation for skin of the present invention In contrast to 60 female volunteers suffering from pigmentation such as dark black, stains, buckwheat, etc., the three groups of statistically equivalent A, B, C groups Separately, Group A uses the emulsion of Example 3 of the present invention, Group B uses the emulsion of Example 4 of the present invention, Group C uses the emulsion of Comparative Example 1 for 3 months on the face, respectively. received. The improvement effect on pigmentation after 3 months was evaluated by visual observation, and comparison between groups was performed. The results are shown in Table 3. In addition, the effective rate was considered to be effective when an effect slightly more effective was recognized.

  From the results of Table 3, it was confirmed that the emulsions of the present invention (emulsions of Examples 3 and 4) had a remarkable pigmentation-inhibiting action.

  In the same manner as in Example 3, an emulsion that is an external preparation for skin of the present invention was prepared according to the formulation shown below. That is, the components (A) were mixed and heated to 80 ° C. On the other hand, each component of (B) was heated to 80 degreeC. The mixture of (B) was added to the mixture of (A) and stirred to emulsify. Further, (C) was added to neutralize, and then the mixture was stirred and cooled to 35 ° C. to obtain the emulsion of Example 5.

(A)
Behenyl alcohol 0.5% by mass
Cetyl isooctanoate 2.0 mass%
Squalane 8.0 mass%
Dimethicone 2.0 mass%
Sorbitan sesquistearate 1.5% by mass
POE (45) stearic acid 1.0 mass%
Cetyl stearate 0.5 mass%
Behenic acid 0.5% by mass
(B)
1,3-butanediol 5.0% by mass
Glycerin 5.0% by mass
1,2-pentanediol 5.0% by mass
Pure water 50.0% by mass
Amorisin 2.0 mass%
Dipotassium glycyrrhizinate 0.1% by mass
(C)
Pure water 16.3 mass%
Potassium hydroxide 0.6 mass%

According to the formulation shown below, each component was mixed to obtain a lotion of Example 6.
Polyethylene glycol (1500) 2.5 mass%
1,3-butanediol 8.0% by mass
Glycerin 10.0% by mass
Methylparaben 0.2 mass%
Sodium hydrogen phosphate 0.1% by mass
Amoricin 0.001% by mass
1,2-hexanediol 0.3% by mass
Pure water 78.899 mass%

Claims (11)

A skin external preparation characterized by containing an amorisin represented by the following chemical formula and / or a salt thereof.

The skin external preparation according to claim 1, which is used for suppressing melanin production. The external preparation for skin according to claim 1 or 2, characterized in that it contains 0.0001 to 5% by mass of amoricin based on the total external preparation for skin. An external preparation for skin, comprising an extract of a leguminous plant belonging to the genus Leguminosae containing amorisin and / or a salt thereof, with a polar solvent. The external preparation for skin according to claim 4, wherein the extract with the polar solvent is an extract with ethanol, hydrous ethanol, polyhydric alcohol, hydrous polyhydric alcohol. The external preparation for skin according to claim 4 or 5, wherein the extract with the polar solvent contains 0.001 to 0.05 mass% of amorisin. It is a cosmetic, The skin external preparation in any one of Claims 1-6 characterized by the above-mentioned. The external preparation for skin according to any one of claims 1 to 7, which is a quasi-drug. Furthermore, the skin external preparation in any one of Claims 1-8 containing 1 type, or 2 or more types selected from glycyrrhizic acid, its salt, and alkyl glycyrrhetinate. The content of one or more selected from the glycyrrhizic acid and salts thereof, and alkyl glycyrrhetinate is 0.05 to 0.5 mass% with respect to the entire skin external preparation, The skin external preparation of Claim 9. The skin external preparation according to claim 9 or 10, wherein the preparation is for melanin production suppression and anti-inflammatory.