JPH1017455A - Agent for inhibiting activity of tyrosinase and cosmetic - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a tyrosinase activity-inhibiting agent capable of strongly inhibiting the tyrosinase activity related to the production of melamine by compounding chlorophorin. SOLUTION: This tyrosinase activity-inhibiting agent contains >=0.0001wt.% of chlorophorin existing in the wood of Chlorophora excelsa as an active ingredient. A beautifully whitening agent containing the tyrosinase activity-inhibiting agent as an essential ingredient can be prepared. The tyrosinase activity- inhibiting agent is added to the cosmetic in an amount of >=0.0001wt.%, preferably 0.0001-10wt.%, as the chlorophorin of active ingredient. Especially, the further addition of a known beautifully whitening agent, a wrinkle-preventing agent, a humectant or their mixture to the cosmetic can synergistically improve the effect. The wood of the Chlorophora excelsa is not toxic, because of being eatable, and can give a wide range of cosmetics, because of being stably added to the cosmetics.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a tyrosinase activity inhibitor effective for the prevention or treatment of skin spots, freckles and the like, and to a cosmetic using the inhibitor.

[0002]

2. Description of the Related Art Conventionally, various substances have been proposed for inhibiting melanin production by inhibiting tyrosinase activity for the purpose of preventing or treating skin spots and freckles. Specifically, vitamin C, hydroquinone, kojic acid, thiol-based compounds, and various animal and plant extracts are known, and cosmetics containing these have been developed and commercialized. However, since vitamin C, hydroquinone, and kojic acid have high polarity, it is necessary to consider this point when blending as cosmetics. Further, the thiol-based compound has a problem in stability when blended into cosmetics. Furthermore, there are few animal and plant extracts that have been proposed so far that have sufficient tyrosinase activity inhibiting ability.

On the other hand, chlorophorin is a mulberry family (Chloro
phora excelsa) is known to exist in the wood,
It has not been known that chlorophorin has the ability to inhibit tyrosinase activity.

[0004]

Accordingly, an object of the present invention is to provide a tyrosinase activity inhibitor which strongly suppresses tyrosinase activity involved in melanin production.
Another object of the present invention is to provide a cosmetic that suppresses tyrosinase activity to suppress melanin production and exhibits a whitening effect and the like.

[0005]

According to the present invention, there is provided a tyrosinase activity inhibitor containing chlorophorin as an active ingredient. Further, according to the present invention, there is provided a cosmetic comprising the tyrosinase activity inhibitor.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail. The tyrosinase activity inhibitor of the present invention contains chlorophorin represented by the following structural formula as an active ingredient.

[0007]

Embedded image

[0008] Chlorophorin is a mulberry family ( Chlorophora).
excelsa ) is known to be obtained by extracting the powder or shavings of this material with ether at a warm temperature, removing the solvent, and recrystallizing the material. For example, Artocar
pus ) can be purified and isolated from a solvent extract of breadfruit ( Artocarpus incisus ) belonging to the same species .

The tyrosinase activity inhibitor of the present invention may contain the above-mentioned active ingredient.
0001% by weight or more.

The cosmetic of the present invention contains the tyrosinase activity inhibitor as an essential component, and preferably exhibits a melanin production inhibitory action based on the tyrosinase activity inhibitory action,
It can be used as a whitening cosmetic. The compounding ratio of the tyrosinase activity inhibitor to the cosmetic is such that chlorophorin as an active ingredient is 0.0001% by weight or more, particularly 0.0001% by weight.
It is desirable that the content be 20 to 20% by weight, more preferably 0.0001 to 10% by weight.

[0011] In addition to the tyrosinase activity inhibitor, various materials can be added to the cosmetic of the present invention according to the purpose. Particularly conventionally known whitening agents, wrinkle preventives,
By blending a humectant or a mixture thereof, the desired effect can be synergistically improved.

Examples of the whitening agent include kojic acid, ascorbic acid, hydroquinone, thiol compounds, derivatives thereof, extracts of animals and plants containing them, and mixtures thereof. As the humectant, for example, glycerin, propylene glycol, 1,
Polyhydric alcohols such as 3-butylene glycol, sorbitol, mannitol, polyethylene glycol and dipropylene glycol; NMF components such as amino acids, sodium lactate and sodium pyrrolidone carboxylate; hyaluronic acid; collagen; elastin; chondroitin sulfate; fibronectin; ceramides A heparin-like substance; a water-soluble polymer substance such as chitosan; a mixture thereof; The blending ratio when blending the whitening agent, anti-wrinkle agent or humectant is preferably 0.001 to 1000 of the active ingredient of the tyrosinase activity inhibitor.
It is desirable to mix in a double amount, particularly preferably in a range of 0.005 to 500 times.

The cosmetics of the present invention include fats and oils, surfactants, alcohols, and the like generally incorporated according to the type of cosmetics.
Fatty acids, preservatives, bactericides, thickeners, antioxidants, pigments, fragrances, water-soluble polymers, ultraviolet absorbers, chelating agents,
Other components such as a pH adjuster, a buffer, and purified water can be appropriately blended.

Examples of the thickener include sodium alginate, xanthan gum, aluminum silicate, quince seed extract, tragacanth gum, starch and other natural high molecular substances; methylcellulose, soluble starch, hydroxyethylcellulose, carboxymethylcellulose, soluble starch, cationized Semi-synthetic polymer substances such as cellulose; synthetic polymer substances such as carboxyvinyl polymer and polyvinyl alcohol; and mixtures thereof. As the preservative, benzoate, sorbate, dihydroacetate, p-hydroxybenzoate,
Examples thereof include 2,2,4'-trichloro-2'-hydroxydiphenyl ether, 3,4,4'-trichlorocarbanide, benzalkonium chloride, and ethanol. Examples of the antioxidant include dibutylhydroxytoluene, butylhydroxyanisole, vitamin E, propyl gallate and the like. Examples of the ultraviolet absorber include 4-methoxybenzophenone, octyldimethylparaaminobenzoate, ethylhexylparamethoxycinnamate, titanium oxide, kaolin, and talc. Examples of the chelating agent include ethylenediaminetetraacetate, pyrophosphate, hexametaphosphate, citrate, tartaric acid, gluconic acid and the like. Examples of the pH adjuster include sodium hydroxide, potassium hydrogen phosphate, potassium carbonate, citric acid and the like.

The cosmetic of the present invention can be prepared as a drug, a quasi-drug, or a cosmetic by mixing the essential components and other components as necessary for various purposes. Specifically, lotions, emulsions, creams,
Packing agents, skin cleansing agents, haptics, plasters, pastes, ointments, essences, gels, shampoos, rinses, powders, foundations, lotions, facial cleansers, hair tonics, hair tonics, bath preparations, etc. it can. In this case, as the other components, in addition to the above-described components, components conventionally used for such applications can be appropriately selected and blended.

[0016]

Industrial Applicability The tyrosinase activity inhibitor of the present invention has no toxicity and is not involved in tyrosinase activity involved in melanin production since chlorophorin, which is also found in edible mulberry wood, is used as an active ingredient. Can be strongly suppressed. In addition, the cosmetic of the present invention contains the tyrosinase activity inhibitor as an essential component, so that by suppressing tyrosinase activity, melanin production can be suppressed and a whitening effect can be expected, and the active ingredient is contained in the cosmetic. Can be stably compounded, so that various cosmetics can be obtained over a wide range.

[0017]

The present invention will be described in more detail with reference to the following examples.
The present invention is not limited to these.

Example 1 37 kg of wood of breadfruit (A. incisus) was finely ground with a grinder to obtain breadfruit powder. The obtained bread tree powder was immersed in ether and kept at room temperature for 24 hours. Then, after stirring, filtration was performed to obtain an ether extract. Solvent and altocarpine (artcarpi) were obtained from the obtained ether extract.
n) was distilled off to obtain a mother liquor. The obtained mother liquor was treated and fractionated by HPLC several times, and fractions showing tyrosinase inhibitory activity were selected. A compound contained in the obtained active fraction,
LC-HRFABMS (liquid chromatography-high re
solution fast atom bombardment mass spectometry), it was estimated to be C ₂₄ H ₂₈ O ₄ ,
When analyzed by MR, it was identified as chlorophorin.
The isolated chlorophorin was subjected to the following tyrosinase activity inhibition test and the like.

Tyrosinase activity inhibition test 2.0 ml of 0.1% L-tyrosine solution (Mcilvaine buffer, pH 6.8), 0.2 ml of dimethyl sulfoxide solution containing chlorophorin as a sample serial dilution, and mushroom-derived Tyrosinase (Sigma) 12
Mix 50 ml of 50 unit / ml buffer,
For 10 minutes. Then, the absorbance was measured at 475 nm to determine the IC ₅₀ for the control. As a result, the IC ₅₀ of chlorophorin was 26 ppb, indicating that kojic acid (IC ₅₀ = 1) exhibiting a known tyrosinase inhibitory activity.
230 ppb).

[0020] A method for B-16 mouse melanoma cells
Lanin production inhibition test B-16 mouse melanoma cells were treated with M containing 10% FBS.
It was adjusted to 1 × 10 ⁵ cells / 5ml in EM medium and 5ml seeded in T-25 flasks. Then, the cells were cultured at 37 ° C. for 24 hours in a CO ₂ incubator adjusted to 5% CO ₂ .
After confirming that the cells had completely adhered, a sample dissolved in dimethyl sulfoxide was added to a final concentration of 100 ppm. The culture period was 6 days, and the medium was replaced on the third day. After completion of the culture, the cells were collected by trypsin treatment, and the degree of whitening and the degree of cell culture were visually determined according to the criteria shown in Tables 1 and 2. Table 3 shows the results. From the results in Table 3, chlorophorin is not cytotoxic,
A melanin production inhibitory effect sufficient for practical use was obtained.

[0021]

[Table 1]

[0022]

[Table 2]

[0023]

[Table 3]

Inhibitory effect of guinea pig on melanin production
The back of the test brown guinea pig was shaved and shaved with a clipper.
Next, cover a 1 × 1.5 cm rectangular hole with aluminum foil with six holes, and irradiate with UV-B lamps (trade name: FL40S / BLB (manufactured by Toshiba Corporation): 6, ² mW / cm ² in total). Quantity 1
(J / cm ² ) three times a week and repeated for two weeks to form pigmentation. The guinea pigs were left for one week to stabilize pigmentation. 4
From the week, each sample 1 once a day, 5 times a week
2.5 μl was applied to the pigmented site for 4 consecutive weeks. Before application, 1, 2, 3 weeks and 4 weeks after application, the back was photographed, and the color difference before and after application was measured with a colorimeter (trade name: RC-100, manufactured by Minolta). Measure,
The ΔL value was calculated from the difference to determine the blackening of the skin color. Table 4 shows the results. From the results in Table 4, it was found that chlorophorin significantly suppressed melanin production.

[0025]

[Table 4]

Example 2, Comparative Example 2 4.0 parts by weight of stearic acid, 3.0 parts by weight of cetyl alcohol, 1.0 part by weight of stearyl alcohol, 6.5 parts by weight of liquid paraffin, 10.0 parts by weight of petrolatum, sorbitan 1.5 parts by weight of monostearate and 3.0 parts by weight of polyoxyethylene monostearate (25EO) were dissolved by heating. Next, 0.5 parts by weight of chlorophorin prepared in Example 1, 5.0 parts by weight of 1.3-butylene glycol, 0.1 parts by weight of potassium hydroxide, and 65.4 parts by weight of purified water were added to the heat-dissolved solution. And then cooled to obtain a cream. For comparison, a cream was obtained in which the amount of purified water was 65.9 parts by weight without using chlorophorin. Each of the obtained creams was used by 10 female panels aged 20 to 30 years twice a day in the morning and before bedtime for one month,
Thereafter, the degree of improvement in skin spots and freckles was evaluated. The evaluation was made by visual judgment as effective 10 points, slightly effective 5 points, and invalid 0 points. As a result, the average score was 7.0 in Example 2 containing the chlorophorin prepared in Example 1, and 0 in the comparative cream.

Example 3, Comparative Example 3 0.5 parts by weight of chlorophorin, 0.5 parts by weight of polyoxyethylene cetyl ether, 2.0 parts by weight of propylene glycol, 0.1 part by weight of 1-menthol prepared in Example 1 Parts and 96.9 parts by weight of purified water were uniformly stirred to obtain a lotion. As a comparison, a lotion was obtained in which the amount of purified water was 97.4 parts by weight without using the chlorophorin prepared in Example 1. Each of the obtained lotions was used by 10 panelists in the same manner as in Example 2, and the degree of improvement in skin spots and freckles was similarly evaluated. As a result, the average was 6.5 points in Example 3 in which the chlorophorin prepared in Example 1 was blended, and 0 points in the comparative lotion.

Example 4, Comparative Example 4 0.5 parts by weight of chlorophorin prepared in Example 1, 1.0 part by weight of polyoxyethylene hydrogenated castor oil (60EO), 15.0 parts by weight of ethanol, 1 part by weight , 3-butylene glycol 4.0 parts by weight and purified water 79.5 parts by weight were mixed and stirred, and each component was dissolved to prepare a lotion. As a comparison, a lotion was prepared in which the amount of purified water was 80.0 parts by weight without using the chlorophorin prepared in Example 1. Each of the obtained lotions was used by 10 panelists in the same manner as in Example 2, and the degree of improvement of skin spots and freckles was similarly evaluated. As a result, the average was 8.5 points in Example 4 containing the chlorophorin prepared in Example 1, and 0 points in the comparative lotion.

Example 5, Comparative Example 5 8.0 parts by weight of squalane, 2.0 parts by weight of petrolatum, 0.5 parts by weight of beeswax, 0.8 parts by weight of sorbitan sesquiolate and polyoxyethylene oleyl ether (20 parts by weight)
E. FIG. O. ) 1.2 parts by weight were dissolved by heating. The chlorophorin 0.5 prepared in Example 1 was added to the obtained heat-dissolved solution.
Parts by weight, carboxyvinyl polymer 0.2 parts by weight, propylene glycol 5.0 parts by weight, potassium hydroxide 0.1
After mixing 2.0 parts by weight of ethanol, 2.0 parts by weight of ethanol and 79.7 parts by weight of purified water, the mixture was cooled to prepare an emulsion. For comparison, an emulsion was prepared without using the chlorophorin prepared in Example 1 and using 80.2 parts by weight of purified water. Each of the obtained emulsions was used by 10 panelists in the same manner as in Example 2, and the degree of improvement in skin spots and freckles was similarly evaluated. As a result, the average was 7.0 in Example 5 containing the chlorophorin prepared in Example 1, and 0 in the comparative emulsion.

Example 6 8.0 parts by weight of squalane, 2.0 parts by weight of petrolatum, 0.5 parts by weight of beeswax, 0.8 parts by weight of sorbitan sesquiolate and polyoxyethylene oleyl ether (20 parts by weight)
E. FIG. O. ) 1.2 parts by weight were dissolved by heating. The chlorophorin 0.5 prepared in Example 1 was added to the obtained heat-dissolved solution.
Parts by weight, carboxyvinyl polymer 0.2 parts by weight, propylene glycol 5.0 parts by weight, potassium hydroxide 0.1
After mixing 2.0 parts by weight of ethanol, 2.0 parts by weight of ethanol, 1.0 part by weight of kojic acid and 78.7 parts by weight of purified water, the mixture was cooled to prepare an emulsion.

Example 7 1.0 part by weight of polyethylene glycol monostearate, 2.0 parts by weight of lipophilic glyceryl monostearate,
5.0 parts by weight of olive oil and 2.0 parts by weight of oleic acid were dissolved by heating. 0.5 part by weight of chlorophorin prepared in Example 1, 0.2 part by weight of hydroxyethyl cellulose, 2.0 parts by weight of propylene glycol, 0.1 part by weight of dipotassium glycyrrhizinate and purified water 87 were added to the obtained heat-dissolved solution. After mixing 2 parts by weight, the mixture was cooled to prepare an emulsion.

Example 8 4.0 parts by weight of stearic acid, 3.0 parts by weight of cetyl alcohol, 1.0 part by weight of stearyl alcohol, 6.5 parts by weight of liquid paraffin, 10.0 parts by weight of petrolatum, sorbitan monostearate 1 0.5 parts by weight and 3.0 parts by weight of polyoxyethylene monostearate (25EO) were dissolved by heating. Next, 0.5 parts by weight of the chlorophorin prepared in Example 1, 5.0 parts by weight of 1.3-butylene glycol, 0.1 parts by weight of potassium hydroxide, 0.2 parts by weight of arbutin and After mixing 65.2 parts by weight of purified water, the mixture was cooled to obtain a cream.

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[Procedure amendment]

[Submission date] August 13, 1997

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0018

[Correction method] Change

[Correction contents]

Example 1 37 kg of wood of breadfruit (A. incisus) was finely ground with a grinder to obtain breadfruit powder. The obtained bread tree powder was immersed in ether and kept at room temperature for 24 hours. Then, after stirring, filtration was performed to obtain an ether extract. Solvent and altocarpine (artcarpi) were obtained from the obtained ether extract.
n) was distilled off to obtain a mother liquor. The obtained mother liquor was treated and fractionated by HPLC several times, and fractions showing tyrosinase inhibitory activity were selected. A compound contained in the obtained active fraction,
LC-HRFABMS (liquid chromatography-high re
solution fast atom bombardment mass spectometry), it was estimated to be C ₂₄ H ₂₈ O ₄ ,
When analyzed by MR, it was identified as chlorophorin.
For the isolated chlorophorin, the following tyrosinase activity inhibition test was performed on B-16 mouse melanoma cells.
Melanin production inhibition test and mela using guinea pig
A nin production suppression effect test was performed.

──────────────────────────────────────────────────続 き Continuation of the front page (51) Int.Cl. ⁶ Identification code Agency reference number FI Technical display location A61K 35/78 AED A61K 35/78 AEDD (72) Inventor Kenichi Kakino 2 Moroka, Hakata-ku, Fukuoka City, Fukuoka Prefecture Chome 13-38 Daiwa Corp. No. 305 (72) Inventor Hiroaki Sato 3-1-25 Haradaso 202-3, Yamada, Onojo-shi, Fukuoka Prefecture (72) Inventor Hiroshi Nagafuku 1-10 Chikushi Ekimae-dori, Chikushino-shi, Fukuoka Prefecture City Heim Chikushi 312

Claims (2)

[Claims] 1. A tyrosinase activity inhibitor comprising chlorophorin as an active ingredient. 2. A cosmetic comprising the tyrosinase activity inhibitor according to claim 1.