JP2012102063A - Melanogenesis inhibitor and skin-whitening agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a melanogenesis inhibitor and a skin-whitening agent.SOLUTION: The melanogenesis inhibitor and the skin-whitening agent contain an amide composition having an acetylenic group.

Description

  The present invention relates to a melanin production inhibitor and a whitening agent.

  The amount of ultraviolet rays that reach the epidermis tends to increase year by year due to air pollution and the destruction of the ozone layer, and skin troubles such as skin spots, freckles, and darkness due to ultraviolet rays are increasing accordingly. Ultraviolet rays oxidize tyrosine present in the skin by the action of the tyrosinase enzyme to produce melanin pigment, and when it is produced in excess, it causes skin problems such as spots, freckles, and darkness. As a method for suppressing the production of this melanin pigment and preventing spots, freckles, and darkness, an external preparation for skin containing arbutin (Patent Document 1: Japanese Patent Laid-Open No. 60-16906), L-ascorbic acid and its A skin cosmetic (Patent Document 2: Japanese Patent Laid-Open No. Hei 4-182412) in which a glucose glycoside of L-ascorbic acid which is a derivative is blended has been proposed. Cosmetics containing arbutin (Patent Document 3: JP 2009-67691 A) and whitening agents containing L-ascorbic acid and a glucose glycoside of L-ascorbic acid which is a derivative thereof have a sufficient whitening effect. However, when a component showing a whitening effect is blended in a concentration at which the whitening effect is recognized, a safety problem may occur.

  Moreover, although Undeca-2,4-diene-8,10-diynoic acid isobutyl-amide is known to promote phase 2 protein production, it is not known about whitening or melanin production inhibitory activity (Patent Document 4: International Publication No. 2010/003238).

Japanese Patent Laid-Open No. 60-016906 Japanese Patent Laid-Open No. 04-182412 Japanese Patent Application No. 2009-67691 International Publication No. 2010/003238

  An object of the present invention is to provide a safe and useful melanin production inhibitor and whitening agent.

（化学式１）
式中、R_１，R₂は、それぞれ独立に水素原子または置換されていても良いアルキル基、あるいはR1，R2は一緒になって環を形成してもよい基である。
４．化学式１で示される化合物を含有する美白剤。
５．下記の化学式２で示されるアミド化合物を有効成分とするメラニン産生抑制剤。

（化学式２）
６．化学式２で示されるアミド化合物を含む美白剤。
７．下記の化学式３で示されるアミド化合物を有効成分とするメラニン産生抑制剤。

（化学式３）
８．化学式３で示されるアミド化合物を含む美白剤。 As a result of repeated studies to solve the above problems, the following melanin production inhibitors and whitening agents have become effective.
1. Melanin production inhibitor containing tarragon extract as an active ingredient.
2. Whitening agent containing tarragon extract.
3. The melanin production inhibitor which uses the amide compound shown by following Chemical formula 1 as an active ingredient.

(Chemical formula 1)
In the formula, R ₁ and R ₂ are each independently a hydrogen atom or an optionally substituted alkyl group, or R ₁ and R ₂ are a group that may form a ring together.
4). A whitening agent containing the compound represented by Chemical Formula 1.
5. The melanin production inhibitor which uses the amide compound shown by following Chemical formula 2 as an active ingredient.

(Chemical formula 2)
6). A whitening agent comprising an amide compound represented by Chemical Formula 2.
7). The melanin production inhibitor which uses the amide compound shown by following Chemical formula 3 as an active ingredient.

(Chemical formula 3)
8). A whitening agent comprising an amide compound represented by Chemical Formula 3.

  By using the above melanin production inhibitor and whitening agent in the present invention, a stronger melanin production inhibitor and whitening agent can be obtained. Furthermore, the effect that the melanin production inhibitor, the whitening agent, etc. are safer under active concentration can be acquired.

Scheme for isolating compounds 1 and 2 A graph comparing the amount of melanin produced by an amide compound having an acetylene group of the present invention and a positive control compound

（化学式１）
式中、R_１，R₂は、それぞれ独立に水素原子または置換されていても良いアルキル基、あるいはR1，R2は一緒になって環を形成してもよい基である。

化学式（２）

（化学式３） Hereinafter, embodiments of the present invention will be described in more detail.
The amide compound having an acetylene group contained in the tarragon extract in the present invention is a compound represented by the following chemical formula (1), preferably the following compound 1 represented by the following chemical formula (2) and chemical formula (3): 2.

(Chemical formula 1)
In the formula, R ₁ and R ₂ are each independently a hydrogen atom or an optionally substituted alkyl group, or R ₁ and R ₂ are a group that may form a ring together.

Chemical formula (2)

(Chemical formula 3)

Specifically, for the purpose of searching for a compound having an ability to suppress melanin production applied directly to the human body such as cosmetics, a number of compounds expected to have an ability to suppress melanin production from plant extracts such as tarragon. The compound was screened through a device for measuring melanin production ability, and a compound having high ability to inhibit melanin production was selected. Furthermore, in order to confirm the safety that can be directly applied to the human body, a test for confirming the safety was conducted to search for a compound that has a high ability to suppress melanin production and does not exhibit toxicity at an active concentration.
As a result, from among a large number of compounds, a compound that is superior in safety to melanin production and has safety is identified, and this compound is an amide compound having an acetylene group.
Thus, amide compounds having an acetylene group are contained in plants such as tarragon (Artemisia dracunculus) and can be obtained by isolation and purification from these plant extracts.

  The melanin production inhibitor of the present invention can be used as a whitening agent of the present invention by mixing with various bases and additives for whitening agents when used for developing a whitening effect.

  The whitening agent of the present invention has various known forms and uses, such as emulsified cosmetics for whitening, whitening creams, whitening lotions, whitening oily cosmetics, whitening packs, whitening foundations, whitening oils, etc. Can be used as The whitening agent of the present invention includes a whitening skin external medicine, a whitening quasi-drug, a whitening cosmetic, a whitening facial cleanser, soap, an antiperspirant, a bath agent and the like. The whitening agent of the present invention is not limited to those applied directly to the skin, but may be an oral whitening agent, an injection, or an oral medicine for whitening.

The melanin production inhibitor and whitening agent containing an amide compound having an acetylene group according to the present invention is produced in an arbitrary form such as a solution, a paste, a solid, or a powder according to a commonly used formulation method. can do.
The melanin production inhibitor and whitening agent of the present invention include oils and fats such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, antiseptics Agents, sugars, sequestering agents, polymers such as water-soluble polymers, thickeners, powder components, UV absorbers, UV blockers, moisturizers such as hyaluronic acid, fragrances, pH adjusters, etc. It can be included. Vitamins, skin activators, blood circulation promoters, resident bacteria control agents, active oxygen scavengers, anti-inflammatory agents, other whitening agents, and other medicinal components such as bactericides, and physiologically active components can also be included.

  Examples of the oils and fats include liquid oils such as camellia oil, evening primrose oil, macadamia nut oil, olive oil, rapeseed oil, corn oil, sesame oil, jojoba oil, germ oil, wheat germ oil, triglycerin, glyceryl trioctanoate, and cocoa butter , Coconut oil, hydrogenated coconut oil, palm oil, palm kernel oil, molasses, mollusc kernel oil, hydrogenated oil, hydrogenated castor oil, beeswax, candelilla wax, cotton wax, nuta wax, lanolin, lanolin acetate, liquid lanolin And waxes such as sugarcane wax, liquid paraffin, squalene, squalane, and microcrystalline wax.

  Examples of higher fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and the like.

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

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

  Examples of the anionic surfactant include fatty acid salts such as sodium laurate, higher alkyl sulfates such as sodium lauryl sulfate, alkyl ether sulfates such as POE lauryl sulfate triethanolamine, N-acyl sarcosine acid, sulfosuccinate , N-acyl amino acid salts and the like.

  Examples of the cationic surfactant include alkyltrimethylammonium salts such as stearyltrimethylammonium chloride, benzalkonium chloride, and benzethonium chloride.

  Examples of amphoteric surfactants include betaine surfactants such as alkyl betaines and amide betaines.

  Examples of nonionic surfactants include sorbitan fatty acid esters such as sorbitan monooleate and hydrogenated castor oil derivatives.

  Examples of preservatives include methyl paraben and ethyl paraben.

  Examples of the sequestering agent include edetates such as disodium ethylenediaminetetraacetate, edetic acid, and sodium edetate.

  Examples of polymers include gum arabic, gum tragacanth, galactan, guar gum, carrageenan, pectin, agar, quince seed, dextran, pullulan, carboxymethyl starch, collagen, casein, gelatin, methylcellulose, methylhydroxypropylcellulose, hydroxyethylcellulose, carboxymethylcellulose Examples thereof include vinyl polymers such as sodium (CMC), sodium alginate, carboxyvinyl polymer (such as CARBOPOL), and bentonite.

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

  Examples of the powder component include extender pigments such as talc, kaolin, mica, silica, zeolite, polyethylene powder, polystyrene powder, and cellulose powder, colored pigments such as inorganic white pigment and inorganic red pigment, titanium oxide coated mica, and oxidation. Mention may be made of pearl pigments such as titanium-coated talc and colored titanium oxide-coated mica, and organic pigments such as red 201 and red 202.

  Examples of the ultraviolet absorber include paraaminobenzoic acid, phenyl salicylate, isopropyl paramethoxycinnamate, octyl paramethoxycinnamate, 2,4-dihydroxybenzophenone, and the like.

  Examples of the ultraviolet blocking agent include titanium oxide, talc, carmine, bentonite, kaolin, and zinc oxide.

  Examples of humectants include polyethylene glycol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, diglycerin, polyglycerin, xylitol, maltitol, maltose, sorbitol, glucose, fructose, sucrose, lactose, and chondroitin. Examples thereof include sodium sulfate, sodium hyaluronate, sodium lactate, pyrrolidone carboxylic acid, cyclodextrin and the like.

  Examples of medicinal ingredients include vitamin A oil, vitamin A such as retinol, vitamin B2 such as riboflavin, B6 such as pyridoxine hydrochloride, L-ascorbic acid, L-ascorbic acid phosphate, L-ascorbic acid Vitamin Cs such as monopalmitate, L-ascorbic acid dipalmitate, L-ascorbic acid-2-glucoside, pantothenic acids such as calcium pantothenate, vitamin Ds such as vitamin D2 and cholecalciferol; α- Vitamins such as vitamin E such as tocopherol, tocopherol acetate and DL-α-tocopherol nicotinate can be mentioned.

  Other medicinal ingredients include whitening agents such as glutathione and yukinoshita extract, skin activators such as royal jelly and beech tree extract, blood circulation such as capsaicin, gingeron, cantalis tincture, ictamol, caffeine, tannic acid, and γ-oryzanol. Accelerators, glycyrrhizic acid derivatives, glycyrrhetinic acid derivatives, anti-inflammatory agents such as azulene, amino acids such as arginine, serine, leucine and tryptophan, maltose sucrose condensates of resident bacteria control agents, lysozyme chloride and the like.

  In addition, chamomile extract, parsley extract, wine yeast extract, grapefruit extract, honeysuckle extract, rice extract, grape extract, hop extract, rice bran extract, loquat extract, buckwheat extract, yakuinin extract, assembly extract, merilot extract, birch extract, licorice extract, peony extract , Savonso extract, loofah extract, capsicum extract, lemon extract, gentian extract, perilla extract, aloe extract, rosemary extract, sage extract, thyme extract, tea extract, seaweed extract, cucumber extract, carrot extract, carrot extract, maronier extract, Examples include various extracts such as chammel extract and mulberry extract.

As an oral preparation for whitening, an amide compound having an acetylene group is added as it is, or various nutritional components are added, or it is added to foods and drinks, so that dietary supplements, functional foods, health foods, foods for specified health use Or it can be used as a raw material for ordinary food. For example, suitable auxiliaries such as starch, lactose, maltose, vegetable oil powder, cocoa butter powder, stearic acid and the like can be added.
The effective dose of the amide compound having an acetylene group as an oral whitening agent is appropriately selected and determined according to the age, weight, symptoms, administration route, administration schedule, formulation form, strength of activity of the material, etc. of the subject. For example, 1 to 5000 mg per day is preferable, and 10 to 1000 mg is particularly preferable. This may be divided and administered several times a day.

  As a compounding quantity of the amide compound which has an acetylene group to a whitening agent, although 0.00001-80 mass% is preferable, it is not necessarily restricted to this, 0.0001-80 mass%, Furthermore, 0.001-50 mass% It is good.

[Extraction of amide compound having acetylene group]
Tarragon Artemisia dracunculus whole plant dry matter fractionation was performed according to the scheme of FIG. 100 g of dried dried cherries were extracted with 100% ethanol to obtain 7.1375 g of extract (A). The ethanol extract was subjected to liquid-liquid partition with ethyl acetate and water, and the ethyl acetate layer (B) was subjected to liquid-liquid partition with hexane and 90% methanol. The 90% methanol layer (C) was fractionated on a TSK-GEL G3000S column, and the 50% ethanol layer was separated into three fractions by reverse phase C18 column (Cosmosil 75C18-OPN). Fr. 2 (D) was further divided into 5 fractions by HPLC (Sun Fire ^™ C18 OBD ^™ 10 μm 70% MeOH). Fr. 2 (compound 1) and Fr. 4 (Compound 2) was an amide compound having an acetylene group.

[Identification of amide compound having acetylene group]
Compound 1 of Fr.2 showed peaks of 230 [M + H] ⁺ , 252 [M + Na] ⁺ , 481 [2M + Na] ⁺ from the MS data, and had a molecular weight of 229. Fr. Compound 2 of No. 4 showed peaks of 242 [M + H] ⁺ , 264 [M + Na] ⁺ and 505 [2M + Na] ⁺ from the MS data, and the molecular weight was 241. Moreover, structural analysis by ¹ H-NMR, ¹³ C-NMR and LC / MS was performed. The shift of ¹ H-NMR in CDCl _{3 of} Compound 1 is δ0.91 (6H, d, J = 7Hz), 1.79 (2H, m), 1.96 (1H, s), 3.15 (2H, dd, J = 7 , 7Hz), 5.45 (1H, br.s), 5.78 (1H, d, J = 15Hz), 6.03 (1H, dt, J = 15,7Hz), 6.19 (1H, dd, J = 15,11Hz), 7.17 (1H, dd, J = 15,11Hz) , and the shift in the CDCl ₃ in ^{13 C-NMR δ18.8,20.1 (2C)} , 28.6,31.2,46.9,65.0,68.2,77.2,123.1,129.7139. Undeca2,4-diene-8,10-diynoic acid isobutyl-amide
Was identified.
The shift of ¹ H-NMR of compound 2 in CDCl ₃ is δ1.60 (4H, m), 1.66 (2H, m),
2.00 (1H, s), 3.52,3.64 (each 2H, m), 6.04 (1H, m), 6.25 (1H, dd, J = 10,15Hz), 6.33 (1H, dd, J = 15,7Hz), 7.24 (1H, dd, J = 10, 15 Hz), and the shift in CDCl ₃ in ¹³ C-NMR is δ18.8, 24.6 (2C), 26.1 (2C), 31.3,43.0br, 47.0br, 65.0,
65.4,68.2,77.2,119.9,130.3,138.5,141.9,165.4, which was consistent with the shift in Reference 2, so 1-Piperidin-1y-l-undeca-2,4-diene-8,10-diyn-1 -one identified.
(References)
1. Phyctochemistry.Vol.24 No10.pp.2295-2300.1985
2. Tetrahedron Vo.42.No.10, pp.2707 to 2716,1986

[Melanin production inhibition test]
About the amide compound 1 and the compound 2 which have an acetylene group, the melanin production inhibitory activity was measured by the following tests.

The melanin production inhibitory activity was determined by the following method. Compound 1, compound 2, and arbutin as a positive control were used as specimens, and each was prepared to 10 mg / mL using methanol. B16F1 cells were inoculated with 200 μl of 4 × 10 ⁴ cells / mL using a medium in which 10% FBS was added to DMEM, and were dissolved in 100 nM α-MSH and methanol. Sample Compound 1 and Compound 2 had final concentrations of 0, 1, 10, 30 μg / mL was added, and 0, 1, 3, 10, 30, 100, 300, 1000 μg / mL was added as arbutin, and the cells were cultured for 96 hours under 5% CO ₂ . After completion of the culture, the culture supernatant was measured at a wavelength of 405 nm, and the value of 405 nm when the specimen was dissolved in the medium was subtracted to obtain a quantitative value of the amount of melanin.
The amount of melanin production (%) was determined by the following equation, where A is the quantitative value when only [NLE4, D-PHE7] α-MSH is treated, and As is the quantitative value when the sample is added at each concentration.
Melanin production (%) = As / A × 100

The results are shown in Table 1. The symbols (A) to (D) of each fraction are as described in [Extraction of amide compound having acetylene group]. The EC _{50 for} inhibiting melanin production (concentration of the specimen in the culture medium when the melanin production quantity is 50%) was determined using the measured value of the melanin production quantity at each specimen concentration. The measured values of melanin production at each sample concentration are graphed in FIG.

[Cell viability test (MTT test)]
Cytotoxicity was determined by the following method. Compound 1, compound 2, and arbutin as a positive control were used as specimens, and each was prepared to 10 mg / mL using methanol. B16F1 cells were inoculated with 200 μl of 4 × 10 ⁴ cells / mL using a medium in which 10% FBS was added to DMEM, and were dissolved in 100 nM α-MSH and methanol. Sample Compound 1 and Compound 2 had final concentrations of 0, 1, 10, 30 μg / mL was added, and 0, 1, 3, 10, 30, 100, 300, 1000 μg / mL was added as arbutin, and the cells were cultured for 96 hours under 5% CO ₂ . After the culture, the cells were treated according to the general protocol of the MTT test method, and the cell viability was measured (FIG. 2). [NLE4, D-PHE7] The IC ₅₀ of each sample was calculated with the cell viability when only α-MSH was treated as 100% (Table 1). The cell viability (%) was determined by the following equation, where B was the quantitative value when only [NLE4, D-PHE7] α-MSH was treated, and Bs was the quantitative value when the sample was added at each concentration.
Cell viability (%) = Bs / B × 100

[Safety factor]
The safety factor was obtained by dividing the cell viability IC ₅₀ by the melanin production inhibition EC ₅₀ . The greater the safety factor, the more melanin production can be suppressed with low cytotoxicity. Compound 1 and Compound 2 have an extremely high safety factor and are excellent as a whitening agent. Table 1 shows the safety factor.

Melanin production in the case of using Compound 1 and Compound 2 is reduced to 20.5% when Compound 1 is 10 μg / mL, and 36 when Compound 2 is 10 μg / mL. Reduced to 9%.
On the other hand, when arbutin, which is a positive control, was used, the melanin production amount was almost unchanged at 100 to 98% at concentrations up to 3 μg / mL, and finally the melanin production amount was slightly as 84% when the concentration was increased to 10 μg / mL. Only stays down.
Furthermore, even when the concentration of arbutin was further increased to 30 μg / mL, the production amount of melanin was still 41%, which was higher than that of Compound 1 at 10 μg / mL.
Only when the concentration of arbutin is 30 μg / mL or more, the concentration is the same as when the concentration of compounds 1 and 2 is 10 μg / mL.
According to the results regarding the amount of melanin produced by compound 1, compound 2, and arbutin, compound 1 and compound 2 exhibit sufficient melanin production suppression at a much lower concentration than arbutin, and at the effective concentration. Toxicity was not observed. According to this result, compounds 1 and 2 can exhibit a melanin production inhibitory action superior to arbutin with the same safety as arbutin.

  Below, the formulation example of this invention is shown. Each was produced according to a conventional method.

Formulation Example 1 Whitening Lotion Ingredients Amount (% by mass)
1. Glycerin 10
2.1,3-Butylene glycol 5
3. Glucose 2
4). Ethanol 5
5. Carboxyvinyl polymer 0.02
6). Dipotassium glycyrrhizinate 0.1
7). Sodium hyaluronate 0.001
8). Undeca2,4-diene-8,10-diynoic acid isobutyl-amide 0.1
9. Citric acid 0.05
10. Sodium citrate 0.1
11. Potassium hydroxide 0.01
12 Purified water residue

Formulation Example 2 Whitening Cream Ingredients Amount (% by mass)
1. Stearyl alcohol 6
2. Stearic acid 2
3. Squalane 10
4). Octild decanol 5
5. Olive oil 5
6.1,3-Butylene glycol 8
7). Polyethylene glycol 1500 4
8). POE (25) cetyl alcohol ether 3
9. Glyceryl monostearate 2
10. Tarragon extract 0.1
11. Purified water residue

Formulation Example 3 Whitening Pack Ingredients Amount (% by mass)
1. Polyvinyl alcohol 15
2. Carboxymethylcellulose 5
3.1,3-Butylene glycol 5
4). Ethanol 12
5. Undeca2,4-diene-8,10-diynoic acid isobutyl-amide 0.05
6). POE oleyl alcohol ether 0.5
7). Citric acid 0.02
8). Sodium citrate 0.04
9. Purified water residue

Formulation Example 4 Whitening Tablet Ingredients Amount (% by mass)
1. Undeca2,4-diene-8,10-diynoic acid isobutyl-amide 63
2. Lactose 24
3. Cornstarch 12
4). Guar gum 1

Formulation Example 5 Whitening beverage Ingredients Amount (mass%)
1. Undeca2,4-diene-8,10-diynoic acid isobutyl-amide 10
2. Fructose butter sugar liquid sugar 15
3. Citric acid 10
4). Vitamin C 5
5. Fragrance 1
6). Dye 1
7). Purified water residue

Claims (8)

  Melanin production inhibitor containing tarragon extract as an active ingredient.   Whitening agent containing tarragon extract. The melanin production inhibitor which uses the amide compound shown by following Chemical formula 1 as an active ingredient.

(Chemical formula 1)
In the formula, R ₁ and R ₂ are each independently a hydrogen atom or an optionally substituted alkyl group, or R ₁ and R ₂ are a group that may form a ring together.   A whitening agent containing the compound represented by Chemical Formula 1. The melanin production inhibitor which uses the amide compound shown by following Chemical formula 2 as an active ingredient.

(Chemical formula 2)   A whitening agent comprising an amide compound represented by Chemical Formula 2. The melanin production inhibitor which uses the amide compound shown by following Chemical formula 3 as an active ingredient.

(Chemical formula 3)   A whitening agent comprising an amide compound represented by Chemical Formula 3.

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