WO1993019721A1

WO1993019721A1 - Novel cosmetic

Info

Publication number: WO1993019721A1
Application number: PCT/JP1992/001338
Authority: WO
Inventors: Shinkichi Honda; Yoichiro Takekoshi; Yoichiro Arai
Original assignee: Kyowa Hakko Kogyo Co., Ltd.
Priority date: 1992-03-31
Filing date: 1992-10-15
Publication date: 1993-10-14
Also published as: JP3459065B2

Abstract

A melanin-coated pigment prepared by coating a carrier with a readily alkali-soluble natural melanin and a cosmetic containing the coated pigment. The coated pigment is highly safe and has a highly developed and saturated color tone, and the cosmetic promises to have both ultraviolet absorption and antioxidant effects.

Description

m

New cosmetics

Technical field

The present invention relates to a melanin-coated pigment obtained by coating a carrier with a natural melanin which is easily soluble and a cosmetic containing the same.

Background technology

Melanin is a brown or black pigment and is known to have properties such as ultraviolet absorption.

As a black pigment, it is known that sepia melanin collected from squid ink bags is dispersed in the system as in carbon black and black iron oxide and used in cosmetics.

(JP-A-55-53211).

Further, US Pat. No. 4,961,754 discloses a powder obtained by modifying the surface of an inorganic powder with a synthetic melanin formed by an oxidative polymerization reaction of a compound having an indole skeleton and a cosmetic containing the powder.

Powders modified with synthetic melanin polymerized on the powder surface are used for cosmetics such as skin irritation caused by oxidation catalysts such as low molecular weight compounds, free monomers remaining in the synthesis process, and potassium iodide and hydrogen peroxide. There is a problem in safety when used as.

Sepia melanin is difficult to isolate and purify because it is insoluble in acids and alcohols and hardly soluble in various solvents, and is difficult to handle as a cosmetic.

When melanin was adsorbed on a carrier, it was found to be usable for coloring purposes as a skin color or gray pigment different from the color of melanin, and was found to be particularly useful for coloring cosmetics.

DISCLOSURE OF THE INVENTION

According to the present invention, a melanin-coated pigment obtained by coating a carrier with a readily soluble natural melanin and a cosmetic containing the pigment are provided.

The carrier to be used is insoluble in water, ash and the like, and can be any powder that can adsorb melanin. For example, inorganic or organic pigments are preferably used.

Inorganic pigments are pigments made of inorganic compounds, for example, titanium dioxide, talc, Sericite, My strength, Titanium mica, Montmorillonite, Kaolin, Alumina, Magnesium oxide, Cesium oxide, Silica, Kaolin, Muscovite, Parium sulfate, Aluminum silicate, Aluminum magnesium magnesium, Sodium magnesium magnesium , Calcium gayate, iron oxide, calcium carbonate, pumice powder, ultramarine, etc.

The organic pigment is a pigment composed of an organic compound, and for example, various dyes and pigments can be used. In addition, as inorganic or organic powder, resins such as nylon 6, nylon 6.6, nylon powder, silicone branch j, fife cellulose, crystal cellose, and silk powder can be used.

These carriers having a particle size of 0.01 to 100 microns are used singly or in combination.

The readily soluble natural melanin in the present invention refers to easily soluble alkali melanin derived from microorganisms and plants, and a preferred example is oleobasidium.

CAureobasidium), Aspergillus and Streptomyces produced by microorganisms belonging to the genus (hereinafter referred to as microbial melanin). These include, for example, FFM. Microbiology Letters 9_, 237-240 (1980), Transactions 'Obza' Pretish 'Mycology' Society (Trans. Br. Ycol. Soc.) 70 (3), 453-455 (1978), Molecular 'and' General Genetics (Mol. Gen. Genet.) 200, 422-432 (1985).

The alkali-soluble natural melanin used in the present invention easily dissolves in an alkaline solution such as sodium hydroxide, hydroxylammonium or ethanolamine of 0.01 to 6.0N.

The melanin-coated pigment of the present invention can be produced by applying a known method for adsorbing components in a solution to a carrier, and can be obtained, for example, by the following production method.

The above melanin is dissolved in an alkaline solution such as sodium hydroxide (0.01 to 6.0), τ-oxidizing steam or ethanolamine, and a carrier is added and stirred. Get An acid solution such as 0.01-6.0N hydrochloric acid is added little by little to the colored suspension to be stirred to sufficiently adsorb melanin on the pigment surface. Decreasing the pH to 1-4 makes the supernatant clear. The solution is filtered and washed with 5 to 10 volumes of water 2 to 5 times, and the residue is dried, and the dried material is ground to obtain a powdered melanin-coated pigment of 0.05 to 50 zm. Is obtained.

Alkali-soluble natural melanin is coated on the surface of the carrier at a rate of 0.01 to 50% by weight.

The thus obtained melanin-coated pigment can be used in paints, inks, cosmetics and the like as a pigment for flesh color or gray. Especially when used in cosmetics, the effect of ultraviolet absorption can also be expected.

When using the melanin-coated pigment of the present invention in Icheon, the type of carrier and the amount of melanin coverage can be experimentally changed to obtain a melanin-coated pigment having a desired color. Such experiments can be easily performed by those skilled in the art.

Examples of cosmetics that can use the melanin-coated pigment of the present invention include cosmetics of any dosage form, such as solutions, emulsions, powders, jellies, and solid preparations. Skin cosmetics such as milky lotions, creams, stone ages, and packs, and cosmetics for hair such as lipsticks, foundations, eye shadows, eyeliners, and the like, and W hairstyles such as IE Shamboo and rinses.

The cosmetic of the present invention comprises various components generally used in cosmetics, such as fats and oils, hydrocarbons, waxes, fatty acids, synthetic esters, alcohols, surfactants, thickeners, humectants, and preservatives. Agents, fragrances, pigments, UV absorbers, drugs, water, etc. can be selected and blended according to the desired dosage form.

Examples of oils and fats include jojoba shan, castor ash, olive ash, soybean oil, coconut oil, palm oil, cocoa shoot, mink shoot, turtle shoot, yashishto, fatty acid diethanolamide, sea avatar, etc. .

Examples of hydrocarbons include liquid paraffin, solid paraffin, petrolatum, microcrystalline wax, and squalane.

The waxes include beeswax, mokurou, lanolin, carnauba wax, candelie Larrow, hydrocarbons, etc. are released.

Fatty acids include myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid and the like.

Synthetic esters include isopropyl myristate, isopropyl palmitate, butyl oleate, myristyl myristate, octyldecyl myristate, propylene glycol mono-stearate, myristinole L, apple diisostearyl, glycerin monostearate, and chloride. Distearyl dimethylammonium, acetyltributyl citrate and the like can be mentioned.

Fats, hydrocarbons, α 類 s, fatty acids, and synthetic esters are usually added together in a ratio of 0.01 to 7 O w / v ^.

Examples of the alcohols include ethanol, 1,3-butylene glycol, propylene glycol, lauryl alcohol, ceanol, stearyl alcohol, oleyl alcohol, and denatured alcohol. It is blended in proportions.

Examples of the surfactant include polyoxyethylene (60) -cured castor, sodium laurinolate sulfate, pyroglucamic acid isostearate polyoxetylene glyceryl, sodium alkylbenzene sulfonate, and polyoxyethylene (10) stearyl ether , Dialkylsulfosuccinic acid, cetylpyridinium bromide, n-octadecyltrimethylammonium chloride, monoalkylphosphoric acid, N-amrugulinic acid, sucrose fatty acid ester, glyceryl isostearate, boroxyethylene ( 20) sorbitan monostearate, polyoxyethylene (20) sorbitan monooleate, sorbitan sesquioleate estesole, polyoxyethylene (20) sorbitan monooleate, sorbitanses chioleate Monoglyceride stearate, polyoxyethylene cetyl ether phosphoric acid, sodium polyoxoxylene lauryl ether sulfate, lanoxylin reduced voroxyethylene, polyoxoxylene ethylene tristearate (20) sorbitan, polyoxydiethylene trioleate (2 0) Sorbitan, soybean phospholipids, etc., are added at a ratio of 0.01 to 4 OwM. Examples of the thickener include carboxyvinyl polymer, methylpolysiloxane, dextran, carboxymethylcellulose, carrageenan, hydroxymethylcellulose, butyl resin emulsion, polyacrylate emulsion, and the like. It is blended at a ratio of l to 5 w / w%.

Examples of humectants include glycerin, propylene glycol, 1,3-butylene glycol, pyrrolidone carboxylic acid, L-acid, hyaluronic acid, collagen, N-acetyl-L-glutamine, triethanolamine, cunic acid, and cunic acid. Sodium and the like, and are blended at a ratio of 0.01 to 30 w / w%.

Examples of preservatives include benzoic acid, salicylic acid, dehydroacetic acid and salts thereof, phenols such as paraoxybenzoic acid ester, 2,4,4'-trichloro- 1'-hydroxydiphenyl ether, and 3-trifluoro. Methyl 1,4'-dichlorocarnox chloride and the like are included, and are blended at a ratio of 0.01 to 0.5 w / w.

As the fragrance, a fragrance usually used in cosmetics can be used.

Pigments added to cosmetics include titanium dioxide, talc, sericite, myriki, titanium mica, montmorillonite, kaolin, alumina, magnesium oxide, cesium oxide, silica, kaolin, muscovite, barium sulfate, aluminum gayate, Aluminum magnesium silicate, sodium magnesium gayate, calcium gayate, iron oxide, calcium carbonate, pumice powder, ultramarine, nylon 6, nylon 6.6, nylon powder, cellulose, crystalline cellulose, silk powder, etc. It is blended at a ratio of 0.01 to 90 w / w%.

UV absorbers include diisopropyl cinnamate, methyl diisopropyl cinnamate, mono-2-diethyl methoxycinnamate glyceryl hexylate, paraaminobenzoic acid, ethyl paraaminobenzoate, and 4-tert-butyl-4'-methoxy. Sidibenzoylmethane, 2-ethylhexyl paradimethylaminobenzoate and the like are included, and are blended at a ratio of 0.01 to 10 W / W%.

Examples of the drug include vitamin E, vitamin C and derivatives thereof, licorice extract, dipotassium glycyrrhizinate, and bracenta liquid, which are incorporated at a ratio of 0.01 to 5 w / w%. The melanin-coated pigment of the present invention is used alone or as a mixture of two or more, and is incorporated in an amount of 0.01 to 90% by weight based on the cosmetic.

When producing a cosmetic containing the melanin-coated pigment of the present invention, a known production method is applied except that part or all of the pigment used in the production of a known cosmetic is replaced with the pigment of the present invention. Can be manufactured.

The microorganism melanin used in the present invention is highly safe and has a wide absorption band in the ultraviolet region. Further, the pigment obtained by coating with the microbial melanin has a high color development and a high color tone, and the cosmetic containing the pigment has an excellent ultraviolet ray preventing effect. BRIEF DESCRIPTION OF THE DRAWINGS

FIG. I shows the survival curve of L929 fibroblast 1 upon irradiation with UVB (wavelength; the same applies to 300 rats or less).

FIG. 2 shows the survival rate by UV irradiation after microbial melanin treatment at each concentration of L929 fibroblasts. In the figure, Δ indicates the 50% survival rate of UVB irradiation, and □ indicates 4% survival rate of UVΒ irradiation.

Figure 3 ①, ②, ③ and ④ are coated with microbial melanin film, respectively 39.6mJ / cni ² from the top of its, the 61.2mJ / cm ^{^2,} 1 04.4mJZcm ² and 212.4mJ cm ² UVB intensity - of The survival rate of L929 fibroblasts by UVBj! Ft irradiation (wavelength; 25 Onm) is shown. In the figure, [] indicates the control and indicates the survival rate. C Best mode for carrying out the invention

Examples, test examples and comparative examples are shown below. The percentages shown in the examples are percentages by weight.

Example 1. Melanin-coated pigment ω preparation

Sutobutomisesu-O over Leo tumefaciens (Streptomyces aureofaciens) (ATCC 10762) pole medium in Erlenmeyer flasks 2 SL volume strain (starch 2.0%, 0.5% peptone, corn steep Rica one 0.9%, CaC0 ₃ 0.1, H5.5 ) The cells were inoculated into 300 ml and cultured at 30 ° C. for 3 days with shaking (at 20 rpm).

The resulting seed culture solution 30 ^ capacity jar fermentans evening production medium in one (starch 5.0, i3. .096, 0.75% lysine _{_{_{hydrochloride, Mg 2 S0 4 ■ 7H 2}}} 0 50 mg /, _{_{CuS0 4 - 5H 2 0 5 0}} mg j2, ZnS0 4 · 7H 2 0 5 0 mgZ ^, CaC0 3 0. 1%, was inoculated into soybean oil 4.0%) 3 0, 1 day 0 2 7 The culture was carried out by aeration and agitation (rotation speed: 4 25 rpm, aeration rate: 1 O ^ Zmin).

Sodium hydroxide (40 g) was added to the obtained culture solution (10 H), and the mixture was centrifuged (at 8,000 rpm, for 30 minutes), and the supernatant was collected. On the other hand, 1N of 0.1N aqueous sodium hydroxide solution was added to the obtained bacterial cells, followed by centrifugation (rotation speed 8, OOOrpi for 30 minutes) to obtain a supernatant. 'The obtained supernatant was combined with the above supernatant, and 150 ml of 10 N hydrochloric acid was added thereto, followed by centrifugation (at 8,000 rpm for 30 minutes). 1N sodium hydroxide 1 was added to the obtained residue, and the mixture was stirred and filtered to obtain a filtrate. 100 ml of 10N hydrochloric acid was added to the filtrate, followed by centrifugation (at 8,000 rpm for 30 minutes). One liter of purified water was added to the obtained residue, and a washing operation including centrifugation (8,000 rpm, 30 minutes) was performed three times to obtain 60 g of crude microorganism melanin.

To 1.4 g of the obtained crude microorganism melanin, 50 ml of 6N hydrochloric acid was added, and the mixture was heated at 110 ° C for 48 hours to carry out a hydrolysis reaction. The reaction solution was dissolved in 100 ml of 6N sodium hydroxide, filtered through a glass fiber filter paper, and 200 ml of 6N hydrochloric acid was added to the filtrate. The precipitate was separated by sedimentation, water 1 was added to the sediment, and the sediment was separated again. The precipitate was freeze-dried to obtain 1 g of purified melanin. 1 g of this purified melanin was uniformly dissolved in 300 g of a 0.1 N aqueous sodium hydroxide solution. Next, 100 g of a carrier (titanium oxide, My power, talc, nylon powder or sericite) was added and uniformly dispersed, and 400 g of 1 N hydrochloric acid was gradually added with stirring. This was filtered, washed with water and dried, and the dried product was pulverized to obtain a 5 m melanin-coated pigment.

Example 2. Preparation of solid foundation

Using the melanin-coated pigment obtained in Example 1, a solid foundation having the following composition was prepared.

(1) Melanin-coated titanium oxide 9.0%

(2) Melanin coating my ability 55.0%

(3) Melanin-coated talc 22.0%

(4) Iron oxide 1.0% (5) Liquid paraffin 6.0%

(6) Lanolin 4.0%

(7) Sorbitan sesquioleate 1.5%

(8) Spice 0.7%

(9) Paraoxybenzoate 0.8%

The components (6) to (9) were uniformly dissolved in the component (5) while mixing the components CI) to (4) well with a blender, followed by pulverization. This powder was molded under pressure to obtain a solid foundation.

Example 3 Preparation of Liquid Foundation

Using the melanin-coated pigment obtained in Example 1, a liquid foundation having the following composition was prepared.

1) Melanin-coated titanium oxide 3.0%

2) Melanin coated my power 2.0%

3) Melanin-coated nylon powder 0.5%

4) Iron oxide 1.0%

5) Liquid paraffin

6) Beeswax 2.0%

7) Carnauba wax 0.2%

8) Squalane 3.0%

9) Boroxydylene (20) Sol! : 'Tanmozoreet_ 0.5%

10) Sorbitan sesquioleate 1.0%

11) Stearic acid monoglyceride 3.96

12) Boroxyethylene cetyl ether phosphoric acid 0.2%

13) 1,3 Butylene glycol 12.0%

14) Carboxyvinyl polymer (1% aqueous solution) 10.0%

15) Triethanolamine 0.1%

16) Fragrance 0.Ζ%

17) Paraoxybenzoic acid ester 0.2% (18) Purified water 58. Q%

Components (1) to (4), (12) and (13) were mixed and uniformly dispersed by a three-roll mill (pigment part). Components (5) to (11), (16) and (17) are heated and dissolved at 80 ° C, and heated to 80 ° C and uniformly mixed (14), (15), (18) And a pigment part, and emulsified and mixed with a homomixer. This was cooled to obtain a liquid foundation.

Example 4. · Preparation of lipstick

Using the melanin-coated pigment obtained in Example 1, a lipstick having the following composition was prepared.

(1) Hydrocarbon wax 5.0%

(2) Candelillaro 2.0%

(3) Glyceryl isostearate 35.0%

(4) Liquid paraffin 45.0%

(5) Melanin-coated titanium oxide 3.0%

(6) Melanin coating my strength 6.7%

(7) Organic pigment (Red No. 3 aluminum lake) 3.0%

(8) Fragrance 0.3%

After mixing the components (4) to (7) and kneading them uniformly with a three-hole mill, add the components (1) to (3) and (8), dissolve by heating until 85 ° C, and stir. When the mixture became uniform by wandering, it was filled into a container to obtain a lipstick.

Example 5. Preparation of eyeliner

Using the melanin-coated pigment obtained in Example 1, an eyeliner having the following composition was prepared.

(1) Ultramarine 2.0%

(2) Melanin-coated talc 4.0%

(3) Melanin coating strength 4.0%

(4) Vinyl acetate resin emulsion 45.0%

(5) Glycerin 5.0%

(6) Boroxydiethylene '(20) Sorbitan monooleic acid ester 1.0%

(7) Carboxymethyl cellulose (10 aqueous solutions) 15.0% (8) Cetyl tributyl citrate 1.0¾

(9) Purified water 22.7%

(10) Fragrance 0.296

(11) Dehydrodic acid 0.1%

Components (5), (6) and (9) were heated and melted, and components (1) to (3) were added thereto and uniformly mixed with a colloid mill. Components (4), (6) to (8), (10) and C11) were added and uniformly dispersed with a homomixer to obtain an airliner.

Example 6. Preparation of shield ink

Using the melanin-coated pigment obtained in Example 1, an eyebrow having the following composition was prepared.

(1) Melanin-coated titanium oxide 3.0%

(2) Melanin-coated talc 22.0%

(3) Melanin coated sericite 22.4%

(4) Melanin coating my power 15.0%

(5) Ultramarine 2.0%

(6) Mokuro 25.096

(7) Vaseline 7.009

(8) Squalane 3.0%

(9) Paraoxybenzoate 0.3%

(10) Vitamin Ε 0.3%

Components (1) to (5) were uniformly mixed with a blender (pigment part). After the components (6) to αο) were dissolved by heating and uniformly stirred, a pigment portion was added and kneaded, and the mixture was molded into a core to obtain an eyebrow ink. Example 7. Preparation of mascara

Using the melanin-coated pigment obtained in Example 1, a mascara having the following composition was prepared.

(1) Melanin coating my power 10.0%

(2) Polyacrylate emulsion 27.0%

(3) Solid paraffin 7.0%

(4) Lanolin 7.0%

Sorbitan sesquioleate 3.0% (6) Paraoxybenzoate 0.3%

(7) Fragrance 0.2%

(8) Purified water 45.5%

(8) was added to component (1), dispersed by a homomixer, and (2) was added thereto and mixed uniformly. The components (3) to (7) were mixed, heated and dissolved at 70 ° C., the mixture was added, and the mixture was emulsified and dispersed with a homomixer to obtain a mascara.

Example 8. Preparation of sunscreen emulsion

Using the melanin-coated pigment obtained in Example 1, a sunscreen emulsion having the following composition was prepared.

(1) Melanin-coated talc 0.5%

(2) soybean phospholipid 0.1%

(3) 1,3-butylene glycol 5.0%

(4) 2-Diethylhexyl p-dimethylaminobenzoate0.1%

(5) Liquid paraffin 2.0%

(6) Glycerin monostearate 1.009

(7) Carboxyvinyl polymer (1% aqueous solution) 10.0%

(8) Triethanolamine 0.1%

(9) Paraoxybenzoate 0.2%

(10) Fragrance 0.2%

(11) Purified water 80.8%

The components (1) to (3) were mixed and uniformly dispersed in a three-neck mill (pigment part). Components (4) to (6) were heated and melted at 80 ° C (Shantou). The components (7) to (11) were heated and dissolved in 8 CTC, and the pigment was added and stirred uniformly (aqueous phase). The aqueous phase was added to Shantou, emulsified and mixed with a homomixer, and then cooled to obtain a sunscreen emulsion.

Example 9. Preparation of sunscreen cream

Using the melanin-coated pigment obtained in Example 1, a sunscreen cream having the following composition was prepared.

Melanin coated sericite 1.0% (2) Beeswax 2.0%

(3) Carnapa 'wax 0.2%

(4) Cetanol 3.0%

(5) Liquid paraffin 3.0¾

(6) 1,3-butylene glycol 5.0%

(7) Tohistearic acid P0E (20) Sorbitan 1.5%

(8) Glycerin monostearate 3.0%

(9) Carboxyvinyl polymer (aqueous solution) 10.0%

(10) Triethanolamine 0.1%

(11) Paraoxybenzoic acid ester 0.2%

(12) Fragrance 0.Z%

(13) Purified water 70.1%

Ingredients (2) to (h) were dissolved by heating to 80 (Shano phase) Ingredients (1) and (8) to (: 13) were heated to 80 ° C and dissolved uniformly, and the O phase was uniformly stirred. The water phase was added to the oil phase, and the mixture was emulsified and mixed with a homomixer and then cooled to obtain a sunscreen cream.

Example 10, Preparation of sunscreen lotion

Using the melanin-coated pigment obtained in Example 1, a sunscreen composition having the following composition was prepared.

(1) Melanin-coated nylon powder 0.5M

(2) Glycerin 1.0%

(3) 1,3-butylene glycol 5.0%

(4) Cuenoic acid 0.1%

(5) Sodium citrate 0.08%

(6) P0EC60) Hardened castor oil 1.0%

(7) Paraoxybenzoic acid ester 0.2%

(8) Fragrance 0.2%

(9) Alcohol 5.0%

(10) Purified water 86.92% Components (1) to (5) and (10) were heated and dissolved at 80 ° C. and stirred uniformly (aqueous phase). Ingredients (6) to (9) were heated and dissolved at 80 ° C (Shanghai). The aqueous phase was added with uniform stirring, and after cooling, a sun protection composition was obtained.

Example 11. Preparation of sunscreen cream

Using the melanin-coated pigment obtained in Example 1, a sunscreen cream with the following cord was prepared.

(1) Melanin-coated titanium oxide 2A96

(2) Melanin coated talc 1.2%

(3) Stearic acid 1.0%

(4) Seinor 0.5%

(5) Sea avatar '' 0.05%

(6) Castor 0.7%

(7) Methyl polysiloxane 1.0%

(8) Liquid paraffin 1.0%

(9) Natural vitamin E 0.05%

(10) Pro-Shan Shan type glyceryl monostearate 1.0%

(11) Trioleic acid P0E (20) Sorbitan 0.2%

(12) Soy phospholipid 0.2%

(13) Paradimethylami / 2-1 butylhexyl benzoate 3.0%

(14) 4-tert-butyl-4'-methoxydibenzoylmethane 2.0%

(15) Trier evening nolamine 0.7%

(16) Paraoxybenzoate 0.2%

(17) 1,3-butylene glycol A.2%

(18) Carboxyvinyl polymer 0.2%

(19) Denatured alcohol 7.0%

(20) Collagen 0.05%

(21) N-Acetyl -L- Guru Yu Min 0.01%

(22) Plant (hamamelis) extract 0.7% (23) Purified water 72.64%

Components (1), (2), (6), (11), (12) and (17) were mixed and uniformly dispersed by a three-roll mill (pigment part). Components (3) to (5),) to (10), (13) and (14) were heated and dissolved at 80 ° C (oil phase). Components (15) to (18), (21) and (23) were heated and dissolved at 80 ° C, and the pigment was added and stirred uniformly (7K phase). Hydration was added to Shantou, and the mixture was emulsified and mixed with a homomixer, then cooled, and ingredients (19), (20) and (22) were mixed to obtain a sunscreen cream.

Test example 1

Hereinafter, the ultraviolet absorption effect of the microorganism melanin on ultraviolet irradiation will be described.

1) Survival curve of L9 29 fibroblasts by ultraviolet (UVB) irradiation

With 6-well microplate (manufactured by Iwaki Glass) in fetal bovine serum 1 0% the added MEM medium (Nissui Pharmaceutical Co., Ltd.) 2 ml, consisting of murine L 9 2 9 fibroblasts 2 xlO ⁴ cells / well And cultured at 37 ° C. for 3 days in a 5% CO ₂ incubator. After culturing, remove the MEM medium and remove PBS (-) [Composition: sodium chloride 8,000 ing, potassium chloride 200m & anhydrous sodium monohydrogen phosphate 1,150mg, anhydrous potassium dihydrogen phosphate 200mg, purified water 1 2 (pH 7.5), the same applies to the following) Wash the cells with (Takara Shuzo), dispense 1 ml of PBS (-), and change the intensity of UVB (wavelength; 300 images, the same applies hereinafter). Irradiated.

The UV lamp used was FL 2 OS · E (maximum wavelength: 300 nm, manufactured by Toshiba Corporation), and the UV B5 degree was measured with an ultraviolet intensity meter UVR-1 (light receiving unit UVR-25, Tokyo Optical Machinery). It was measured.

PBS (I) after removal, new MEM medium was added 2 ml, in 5% C0 ₂ incubator base one evening one, the protein content was measured by 3 days of culturing at at 37, Ru good to UVB irradiation cell The survival rate was calculated from the following equation.

To determine the amount of protein, cells were suspended from the plate by treatment with 0.05% trypsin, and sonicated for 30 seconds in a microtube (1.5ral, manufactured by Eppendorf) in the presence of 1 ml of PBS (-). After centrifugation (9, OOO rpm, 20 minutes), the supernatant was measured using a protein measurement kit (Bio-Rod). Evening mass of cells irradiated with UVB

Survival rate = X 1 00

Protein content of cells without UVB irradiation Figure 1 shows the survival curve of L929 fibroblasts by UVB irradiation.

2) UVB absorption effect of microbial melanin

0.1 g of microbial melanin was dissolved in 100 ml of a 0.1% aqueous sodium hydroxide solution. When completely dissolved, dissolve one PBS tablet (manufactured by Takara Shuzo) in 10 Oml of the solution, adjust the pH to 7 with hydrochloric acid, sterilize with autoclave (121 ° C, 15 minutes), and remove the microorganism melanin phosphate. A buffer (microbial melanin ZPBS) was prepared.

After culturing L929 fibroblasts in the same manner as in 1), wash the cells with PBS (-), and appropriately dilute microbial melanin / PBS with PBS (-) to make 1 ml / well. And irradiated with 50% or 4% viable UVB, respectively.

After washing the cells with PBS (1) and removing PBS (1), add 2 ml of MEM medium again, culture in the same manner as in 1) above, measure the amount of protein, and measure the cell viability by UVB irradiation. Was calculated.

The result is shown in figure 2.

From the above results, it can be understood that the survival rate of the cells by UVB irradiation increases until the melanin concentration reaches 0.025%, and that the survival rate does not decrease much thereafter.

3) UVB absorption effect of microbial melanin film

Dissolve 0.1 g of microbial melanin in 100 ml of 0.1% aqueous sodium hydroxide solution. When completely dissolved, adjust to pH 7 with hydrochloric acid, sterilize with autoclave (121 15 minutes), and add 1% microbial melanin. An aqueous solution was prepared.

10 g of a 1% microorganism melanin aqueous solution and 10 g of an acrylic resin (manufactured by Mitsubishi Shanka Co.) were dissolved in ethanol to make the total amount 100. Of these, 10 g was uniformly poured into the lid of a 6-hole microphone plate and dried at 45 ° C under reduced pressure to prepare a 0.1% microbial melanin film. '

After culturing L929 fibroblasts in the same manner as in 1) above, remove the MEM medium, wash the cells with PBS (1), dispense 1 ml of PBS (-) again, and place the microbial melanin film above. Irradiation was performed by changing the intensity of UVB from above. After washing the cells with PBS (1) and removing PBS (1), add 2 ml of MEM medium and culture in the same manner as in 1) above, measure the amount of protein, and determine the survival rate of the cells by UVB irradiation. Was calculated.

As a control, the same operation as above was performed without UVB irradiation, and the cell viability was calculated.

The results are shown in Figure 3.

From the above results, Oyo UVB irradiation in 1 0 4. 4 mJZcm ² - Beauty 2 1 2. In UVB irradiation at 4 MJZcm ² is understood to be improved cell viability or叨Ra. Test example 2

Regarding the safety of the microbial melanin used in the present invention, a human 48 hr closed batch test [Nisshin Gakkai, 80 (5), 301-314 (1970)], a controlled oral administration toxicity test [Ministry of Health and Welfare Pharmaceutical Affairs No. 118 ( (February 15, 1984), conducted guidelines on toxicology tests required for the application for approval of the manufacture (import) of pharmaceuticals (Part 1)] and conducted a skin photosensitization test (Western Skin, Vol. 42, No. 5, No. 5) .

The results are shown in Table 1.

Table I Test name Results

Human 48 hr Closed negative (44 cases) 24 males, 20 females 《Tititetest Single oral dose toxicity test LD ₅₀ 2000 mgZkg or more * 1

Skin photosensitization test Negative * 2

* 1 Test animal; use CD-1 (ICR) mouse

* 2 Test animals: Hartley guinea pigs used.

Uses TCSA (3, 3 ', 4', 5-tetrachloro mouth salicyl alide)

From the above results, it is understood that microbial melanin is highly safe and safe to use as a chemical. Test example 3

With respect to the microbial melanin-coated pigment of the present invention, the type of carrier (pigment) and the color tone of the microbial melanin according to the coating concentration were measured in the same manner as in Comparative Example 1 described later, and represented by L, a, and b values.

The results are shown in Table 2. No. 2

Color Microbial melanin coating concentration

Carrier (90

Key 0.5 1.0 2.0

L 45.1 42.5 40.3

Oxidized titanium a 0 τ η 1

0.ί y.1 a.1

b 10.8 11.3 11.7

L 43.73 41.0 36.29

Cell site a 11.54 11.55 10.97

b 14.47 14.15 13.11

L 40.22 37.28 32.25

Talk a 10.30 9.91 9.89

b 10.86 10.42 10.39

L 42.86 27.87 33.15

Nylon powder a 10.70 11.01 10.85

b 14.51 14.54 14.18 From the above results, it can be understood that the color tone of the pigment coated with the microbial melanin changes depending on the type of the pigment used as the carrier.

Test example 4

The UV protection effect of the cosmetic containing the melanin-coated face of the present invention was represented by an SPF (Sun Protection Factor) value.

The measurement of SPF was carried out in accordance with the Japan Chemical Industry Association of Japan's “SPF measurement method standard”.

That is, the sunscreen cream obtained in Example 11 was used as the SPF measurement sample of the present invention (referred to in Table 3 as sunscreen cream).

Also, of the composition of the sunscreen cream of Example 11, components (1) and (2) were not coated with melanin, a cream was prepared in the same manner as in Example 1I, and a control SPF measurement sample was prepared. (Referred to as control in Table 3).

Table 3 shows the results.

Ψ> 3 tables

From the above results, the cosmetic containing the melanin-coated pigment of the present invention has a very high

It shows an S PF value, and is understood to be excellent in the effect of preventing ultraviolet rays.

Comparative Example 1

The color tone and coloring power of melanin, synthetic melanin, sepia melanin, black iron oxide, and carbon black (hereinafter referred to as black pigment) were compared. That is, color tone and tinctorial strength were measured in a system (dispersion system) in which titanium dioxide and each of the above black pigments were kneaded and dispersed as pigments.

In the dispersion, black pigment 0.04%, titanium dioxide 1.96%, polyvinyl alcohol 2.0% of acrylic resin, 5.09 of acrylic resin, 73.0% of 1,3-butylene glycol and 18.0% of purified water were thoroughly kneaded with a three-roll mill to prepare a uniform paste-like measurement sample.

Among the black pigments, microbial melanin and synthetic melanin were also measured in the coating system.

That is, in the coating system, 2.090 titanium dioxide coated with 2% of microbial melanin or synthetic melanin, 2.0% polyvinyl alcohol, 5.0% acrylic resin, 73.0% 1,3-butylene glycol, and purified water 18.0% was thoroughly kneaded with a three-roll mill to prepare a uniform paste-like measurement sample.

Place a fixed amount of the above sample on a hiding power test paper with a white surface and a black surface, mix it with a Doc Yuichi blade to a film thickness of 0.20, dry it into a film, and then colorimeter (CR-200) Minolta) was used to measure each sample, and the color tone was displayed as L, a, and b values. The coloring power was compared by lightness.

The results are shown in Table 4.

Table 4

L: Lightness [measured with a hiding power test paper (white surface)]

a, b: chromaticity Based on the above results, the melanin showed a color tone close to that of synthetic melanin, and had a warm, natural color, unlike the jet black of iron oxide and Ripbon black. That is, the cosmetic containing the microbial melanin-coated pigment of the present invention has a gentle hue suitable for skin color.

The color tone of pigments coated with titanium dioxide with melanin is characterized by higher color development and higher saturation than when dispersed, and when microbial melanin is added to cosmetics, the coating system is more dispersed. It is understood that the stability of the product is higher than that of. Comparative Example 2

Purified water is added to 0.1 g of microbial melanin, synthetic melanin, or 0.1 mg of sodium hydroxide and 0.1 g of sodium hydroxide to make 100 ml, and then adjusted to pH 7 with hydrochloric acid. A solution, a synthetic melanin solution or a cameranin solution was prepared.

Ethanol was added to 0.1 g of di-CL-tocopherol and dissolved to prepare 100 ml of nato-tocopherol solution.

Purified water was added to 0.1 g of ascorbic acid and dissolved to prepare 100 ml of ascorbic acid solution.

A solution of 80% cumene hydroperoxide (hereinafter abbreviated as CP0) solution 5.02 ni was added with purified water and dissolved to prepare 100 ml of a CP0 solution (264 nmolZml).

As a reaction reagent, using Minaichi LPO (manufactured by Kyowa Medex), CPO reacts with a methylene blue derivative using hemoglobin as a catalyst to produce equimolar methylene blue to form a color system. Microbial melanin, ascorbic acid, The antioxidant activity of microbial melanin, ascorbic acid, (^ -hi-tocopherol, synthetic melanin and icaminine) was measured by adding ^ -α-tocopherol, synthetic melanin or imecamnin to the extent of methylene blue production inhibition.

In other words, after preparing each sample for sample, standard or blind as shown in Table 5, blind absorbance at 675 nm wavelength of each sample after the reaction according to the instruction manual attached to the reagent below. The sample for measurement was used as a control, and the elimination rate of lipid peroxide was calculated by the following equation. Five

As one Αι

Lipid peroxide elimination rate (%) = 1 x 1 0 0

During Astd formula, A s is the sample甩Samburu absorbance, A std absorbance of the standard sample, A _B represents the absorbance of Samburu for blind.

Table 6 shows the elimination rate of the lipid peroxides at a concentration of 0.014% for the microorganisms melanin, ascorbic acid, sodium tocopherol, synthetic melanin, and imagenine.

Table 6

That is, the antioxidant ability of microorganisms such as melanin, ascorbic acid, d-α-tocopherol, synthetic melanin and icaminin can be represented by the lipid peroxide elimination rate. From the above results, it is understood that the microorganism melanin used in the present invention has an antioxidant ability. Availability of SJi

According to the present invention, a melanin-coated cosmetic having high safety, high color development, and high color tone and a cosmetic containing the same are provided.

'Melanin-coated pigments can be used in paints, inks, cosmetics, etc. as flesh-colored or gray pigments. In particular, UV absorption and antioxidant food can be expected to be effective when used in cosmetics.

Claims

Claim scope

(1) Melanin-coated pigments obtained by coating a carrier with alkali-soluble natural melanin.

(2) The melanin-coated pigment according to claim 1, wherein the carrier is an inorganic pigment or an organic pigment.

(3) Carriers are titanium dioxide, Yunorek, sericite, Myriki, titanium mica, montmorillonite, kaolin, alumina, magnesium oxide, cesium oxide, silica, kaolin, muscovite, barium sulfate, aluminum gait, and gay acid. 3. The melanin-coated pigment according to claim 2, which is selected from aluminum magnesium, sodium magnesium gayate, calcium gayate, iron oxide, calcium carbonate, pumice powder and ultramarine blue.

(4) The melanin-coated pigment according to claim 2, wherein the carrier is selected from nylon 6, nylon 6.6, nylon powder, silicone resin, cellulose, crystalline cellulose, and silk powder.

(5) The natural melanin according to claim 1, wherein the melanin is produced by a microorganism belonging to a genus selected from the genus Aureobasidium, Aspergillusper and Streptomyces. Melanin-coated pigment.

(6) Cosmetics containing the melanin-coated pigment according to (1), (2), (3), (4) or (5).