JPH10265363A - Tyrosinase inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a tyrosinase inhibitor capable of preventing and improving pigmentation after sunburn, spot, freckle, chloasma, etc., having inhibitory action on tyrosinase activity by including an extract of Uncaria gambir Roxb. SOLUTION: This inhibitor comprises an extract of Uncaria gambir Roxb., an evergreen bush of creeping nature, as an active ingredient. The amount of the active ingredient added is 0.0005-20 wt.%, preferably 0.001-10 wt.% calculated as a dried substance in the total of an external preparation. The medicine is properly formulated with components useful in an external preparation for skin such as cosmetic or medicine and prepared into an ointment, cream, milky lotion, lotion, pack, bathing agent, etc. The extract is obtained by immersing leaf, stem, flower, bark, seed, fruit, the whole plant, etc., of Uncaria gambir Roxb., in an extraction solvent such as ethanol, or heating under reflux, filtering and concentrating the filtrate. The inhibitor has excellent effects on pigmentation after sunburn, spot, freckle, color lightening such as chloasma and skin lightening and is excellent in safety.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a tyrosinase inhibitor having a tyrosinase activity inhibitory effect which is effective in preventing and improving pigmentation, spots, spots, freckles, liver spots, etc. after sunburn by incorporating an extract of Acacia catechu. .

[0002]

2. Description of the Related Art Although there are some unclear points about the mechanism of the occurrence of skin spots and the like, melanin pigments are generally formed due to hormonal abnormalities and stimulation of ultraviolet rays from sunlight. It is believed to be abnormally deposited within. This melanin pigment, which causes skin coloring, is produced in melanin-producing granules (melanosomes) in melanocytes (melanocytes) between the epidermis and dermis,
The generated melanin diffuses to adjacent cells by osmosis. The biochemical reaction in this melanocyte is estimated as follows.

That is, tyrosine, which is an essential amino acid, is converted into dopaquinone by the action of the enzyme tyrosinase, which is converted into black melanin via a red pigment and a colorless pigment by an enzymatic or non-enzymatic oxidative action in the process of producing melanin pigment. is there. Therefore, it is important to suppress the action of tyrosinase, which is the first step of the reaction, to suppress melanin production.

[0004]

However, compounds that suppress the tyrosinase action, except for hydroquinone, have very slow onset of their effects, so that the effect of improving skin pigmentation is not sufficient. On the other hand, although hydroquinone is recognized as having an effect, its use is generally restricted because of its sensitizing properties. In order to improve the safety, attempts have been made to use monoesters or alkyl monoethers of higher fatty acids (Japanese Patent Application Laid-Open No. 58-154507), but the esters are decomposed by hydrolytic enzymes in the body. Therefore, it is not always safe, and ethers which do not sufficiently satisfy safety requirements have not been obtained.

[0005]

Means for Solving the Problems In order to solve these problems, the present inventors examined the tyrosinase activity inhibitory effect of various substances. As a result, it was found that the extract of Acacia catechu had the tyrosinase activity inhibitory effect. And completed the present invention. It has been known since ancient times that the extract of Acacia catechu has an antidiarrheal effect, and has been used as an astringent antidiarrheal and intestinal medicine for diarrhea, stool blood, urinary blood or bloody diarrhea. In recent years, reports have been made on the antioxidant action and the hyaluronidase inhibitory action. (JP-A-59-166585, JP-A-2-
No. 11520). However, there has been no report on the tyrosinase activity inhibitory effect of the Acacia catechu extract, and no application to a whitening agent is known. The present inventors have completed the present invention based on the above findings.

[0006] That is, the present invention relates to an ascendant (Uncaria
gambir Roxb.).

Hereinafter, the configuration of the present invention will be described in detail. Asenyaku used in the present invention is a vine evergreen shrub native to and cultivated in India and Southeast Asia. The extract used in the present invention is obtained by immersing or heating and refluxing the leaves, stems, flowers, bark, seeds or fruits, whole plants of an asparagus with an extraction solvent, followed by filtration and concentration. The extraction solvent used in the present invention may be any solvent that is usually used for extraction, and particularly methanol, ethanol, 1,3
-Alcohols such as butylene glycol, aqueous alcohols, and organic solvents such as acetone and ethyl acetate can be used alone or in combination.

The amount of the plant extract in the present invention is 0.0005 to 20.0% by weight, preferably 0.001 to 10.0% by weight, as a dry matter, based on the total amount of the external preparation.
If the amount is less than 0.0005% by weight, the effects of the present invention cannot be sufficiently exerted. Further, even if the content is 10.0% by weight or more, the effect is not so much improved.

Further, the tyrosinase inhibitor of the present invention includes:
In addition to the above essential components, components commonly used in external preparations for skin such as cosmetics and pharmaceuticals, for example, other whitening agents, humectants, antioxidants, oily components, ultraviolet absorbers, surfactants, thickeners, alcohols Classes, powder components, coloring agents, aqueous components, water, various skin nutrients, and the like can be appropriately compounded as necessary.

In addition, sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, caffeine, tannin, verapamil, tranexamic acid and derivatives thereof, and licorice extract Products, glabridine, hot water extract of karin fruit, various crude drugs, drugs such as tocopherol acetate, glycyrrhizic acid and derivatives or salts thereof, vitamin C, magnesium ascorbate phosphate, glucoside ascorbate, arbutin, kojic acid and the like Other whitening agents, sugars such as glucose, fructose, mannose, sucrose, trehalose and the like can also be appropriately compounded.

The tyrosinase inhibitor of the present invention may be any of those conventionally used in external preparations for skin, such as ointments, creams, emulsions, lotions, packs and bath preparations, and the dosage form is not particularly limited.

[0012]

Next, the present invention will be described in more detail by way of examples. Note that the present invention is not limited by this. The compounding amount is% by weight. Prior to the examples, test methods and results of the tyrosinase activity inhibitory effect of the plant extract of the present invention will be described.

Test Methods and Results 1. Preparation of sample (1) Acacia extract extract 50 g of acacia leaves and shoots were immersed in ethanol for 1 week at room temperature, the extract was concentrated, and ethanol extract 2
g was obtained. This extract was dissolved in DMSO at 1%, the concentration was adjusted by diluting this solution, and the following experiment was performed using this.

2. Cell culture method B16 melanoma cultured cells derived from mice were used. 1
0% FBS and theophylline (0.09 mg / ml)
Was cultured in a CO2 incubator (95% air, 5% carbon dioxide) at 37 ° C. in an Eagle MEM medium containing After 24 hours of culture, the sample solution is added to a final concentration (concentration in terms of dry extract) of 10-2 to 10-5% by weight, and the culture is further continued for 3 days, and the tyrosinase activity inhibitory effect is measured by the following method did.

3. Measurement of tyrosinase activity Before measurement, the medium in the wells is removed and washed twice with 100 μl of PBS. 45 μl of 1% Triton-X in each well
(Rohm and Haas product name, surfactant) containing PBS is added. Shake the plate for 1 minute, break the cell membrane well, and use a microplate reader for 475n.
The absorbance at m was measured, and this was taken as the absorbance at 0 minutes. Thereafter, 5 μl of a 10 mM L-DOPA solution was quickly added, and the mixture was transferred to a 37 ° C. incubator and reacted for 60 minutes. The plate was shaken for 1 minute, and the absorbance (475 nm) at 60 minutes was measured. The tyrosinase activity ratio (%) was defined as the ratio of the absorbance difference of the sample with the plant extract to the absorbance difference between the time of 0 minute and the time of 60 minutes in the case of the sample to which no plant extract was added (control). Table 1 shows the results. In addition, as a reference example, the same test as described above was conducted on an ethanol extract of a scallop (Lamiaceae: Subfamily Lamiaceae) already known to have a tyrosinase activity inhibitory action. Table 1 also shows the results.
In the table, "toxic" means that cytotoxicity was observed, and "-" means that no significant difference was observed within 5% of the risk ratio as compared with the control.

[0016]

[Table 1] ───────────────────────────── Test tyrosinase activity rate (%) ───────── ─ ───────────── Concentration (% by weight) 10-5 10-4 10-3 10-2 ──────────────────── ───────── Acacia catechu extract-87 84 58 Limpus extract---55 ─────────────────────────── ──

Hereinafter, examples of the formulation of tyrosinase inhibitors according to the present invention in various dosage forms will be described as examples.

Example 1 Cream (Formulation) Stearic Acid 5.0% by Weight Stearyl Alcohol 4.0 Isopropyl Myristate 18.0 Glycerin Monostearate 3.0 Propylene Glycol 10.0 Acenia Methanol Extract 0.01 Caustic Potassium 0.2 Sodium bisulfite 0.01 Preservatives Appropriate amount Fragrance Appropriate amount Ion-exchanged water Residue (Preparation method) Add propylene glycol, acetic acid methanol extract and caustic potash to ion-exchanged water, dissolve and heat to 70 ° C (water phase ). The other components are mixed, melted by heating and kept at 70 ° C. (oil phase). The oil phase is gradually added to the water phase, and after the addition is completed, the temperature is maintained for a while to cause a reaction. Thereafter, the mixture is uniformly emulsified with a homomixer and cooled to 30 ° C. while stirring well.

Example 2 Cream (Formulation) Stearic acid 2.0% by weight Stearyl alcohol 7.0 Hydrogenated lanolin 2.0 Squalane 5.0 2-Octyldodecyl alcohol 6.0 Polyoxyethylene (25 mol) Cetyl alcohol ether 3.0 Glycerin monostearate 2.0 Propylene glycol 5.0 Acenya yak ethanol extract 0.05 Sodium bisulfite 0.03 Arbutin 3.0 Ethyl paraben 0.3 Perfume Appropriate amount Ion exchange water Residue (Production method) Ion exchange Add propylene glycol to the water,
Heat and maintain at 70 ° C. (aqueous phase). The other components are mixed, melted by heating and kept at 70 ° C. (oil phase). The oil phase is added to the water phase, pre-emulsified, homogenized with a homomixer, and cooled to 30 ° C. with good stirring.

Example 3 Cream (Formulation) Solid paraffin 5.0% by weight Beeswax 10.0 Vaseline 15.0 Liquid paraffin 41.0 Glycerin monostearate 2.0 Polyoxyethylene (20 mol) Sorbitan monolaurate 2 0.0 Soap powder 0.1 Borax 0.2 Acenyacacetone extract 0.05 Acenyacethanol extract 0.05 Sodium bisulfite 0.03 Ascorbic acid glucoside 2.0 Ethylparaben 0.3 Perfume Appropriate amount Ion-exchange water residue (Preparation method) Soap powder and borax are added to ion-exchanged water, heated and dissolved, and maintained at 70 ° C (aqueous phase). The other components are mixed, melted by heating and kept at 70 ° C. (oil phase). The oil phase is gradually added to the aqueous phase while stirring to carry out the reaction. After completion of the reaction, the mixture is uniformly emulsified with a homomixer, and cooled to 30 ° C. while stirring well after emulsification.

Example 4 Emulsion (Formulation) 2.5% by weight of stearic acid Cetyl alcohol 1.5 Vaseline 5.0 Liquid paraffin 10.0 Polyoxyethylene (10 mol) Monooleate 2.0 Polyethylene glycol 1500 0 Triethanolamine 1.0 Asenyak 1,3-butylene glycol extract 0.01 Carboxyvinyl polymer 0.05 (trade name: Carbopol 941, BFGoodrich Chemical company) Aceonyak acetic acid ethyl ester extract 0.01 Sodium bisulfite 0 .01 2-Ethylhexyl paramethoxycinnamate 2.5 Ethylparaben 0.3 Perfume Appropriate amount Ion-exchanged water Residue (Preparation method) Dissolve the carboxyvinyl polymer in a small amount of ion-exchanged water (phase A). Polyethylene glycol 1500 and triethanolamine are added to the remaining ion-exchanged water, dissolved by heating, and kept at 70 ° C. (aqueous phase). The other components are mixed, melted by heating and kept at 70 ° C. (oil phase). The oil phase is added to the water phase to perform preliminary emulsification, the phase A is added, and the mixture is uniformly emulsified with a homomixer. After the emulsification, the mixture is cooled to 30 ° C. while stirring well.

Example 5 Emulsion (Formulation) Microcrystalline wax 1.0% by weight Beeswax 2.0 Lanolin 20.0 Liquid paraffin 10.0 Squalane 5.0 Sorbitan sesquioleate 4.0 Polyoxyethylene (20 mol) ) Sorbitan monooleate 1.0 Propylene glycol 7.0 Asenacyacetone extract 10.0 Magnesium ascorbate phosphate 3.0 Sodium bisulfite 0.01 Ethyl paraben 0.3 Perfume Appropriate amount Ion-exchange water residue (Preparation method) Add propylene glycol to ion exchange water,
Heat and maintain at 70 ° C. (aqueous phase). The other components are mixed, heated and melted and kept at 70 ° C. (oil phase). While stirring the oil phase, the aqueous phase is gradually added thereto, and the mixture is uniformly emulsified with a homomixer. After emulsification, cool to 30 ° C with good stirring.

Example 6 Jelly (Formulation) 95% Ethyl alcohol 10.0% by weight Dipropylene glycol 15.0 Polyoxyethylene (50 mol) Oleyl alcohol ether 2.0 Carboxyvinyl polymer 1.0 (Product name: Carbopol) 940, BFGoodrich Chemical company) Caustic soda 0.15 L-arginine 0.1 Asenjak 50% aqueous ethanol extract 7.0 Magnesium ascorbate 3.0 3.0 Sodium 2-hydroxy-4-methoxybenzophenone sulfonate 0.05 Ethylenediaminetetra Acetate, 3 sodium, 2 water 0.05 methyl paraben 0.2 Appropriate amount of ion-exchanged water Residue (Preparation method) Carbopol 940 is uniformly dissolved in ion-exchanged water, while a 50% ethanolic aqueous extract of asenyak in 95% ethanol Polyoxyethylene (50 mol)
Dissolve the oleyl alcohol ether and add to the aqueous phase. Then, after adding other components, caustic soda, L
-Neutralize with arginine to thicken.

Example 7 Serum (Prescription) (Phase A) Ethyl alcohol (95%) 10.0% by weight Polyoxyethylene (20 mol) Octyldodecanol 1.0 Pantothenyl ethyl ether 0.1 Aceniac methanol extraction Product 1.5 Ascorbic acid glucoside 4.0 Methyl paraben 0.15 (B phase) Potassium hydroxide 0.1 (C phase) Glycerin 5.0 Dipropylene glycol 10.0 Sodium bisulfite 0.03 Carboxyvinyl polymer 0.2 (Product name: Carbopol 940, BFGoodrich Chemical company) Purified water residue (Preparation method) Dissolve A phase and C phase uniformly, and add A to C phase
Add phases and solubilize. Next, after adding the phase B, filling is performed.

Example 8 Pack (Formulation) (Phase A) Dipropylene glycol 5.0% by weight Polyoxyethylene (60 mol) hydrogenated castor oil 5.0 (Phase B) Asenyaku methanol extract 0.01 Placenta extract 0.05 Olive oil 5.0 Tocopherol acetate 0.2 Ethyl paraben 0.2 Fragrance 0.2 (Phase C) Sodium bisulfite 0.03 Polyvinyl alcohol 13.0 (Saponification degree 90, polymerization degree 2,000) Ethanol 7 0.0 purified water residue (Preparation method) A phase, B phase, and C phase were each dissolved uniformly,
Add phase B to phase and solubilize. Next, this is added to the C phase and then filled.

Example 9 Solid Foundation (Formulation) Talc 43.1 wt% Kaolin 15.0 Sericite 10.0 Zinc White 7.0 Titanium Dioxide 3.8 Yellow Iron Oxide 2.9 Black Iron Oxide 0.2 Squalane 8 0.0 Isostearic acid 4.0 POE sorbitan monooleate 3.0 Isocetyl octoate 2.0 Acenyac ethanol extract 0.5 Licorice extract 1.0 Preservatives Appropriate amount Fragrance Appropriate amount (Production method) talc-black iron oxide powder The components are thoroughly mixed in a blender, and the oily components of squalane-isocetyl octanoate, an acenyac ethanol extract, a preservative, and a fragrance are added, kneaded well, and the mixture is filled into a container and molded.

Example 10 Emulsion type foundation (cream type) (prescription) (powder part) Titanium dioxide 10.3% by weight Sericite 5.4 Kaolin 3.0 Yellow iron oxide 0.8 Bengala 0.3 Black iron oxide 0.2 (oil phase) Decamethylcyclopentasiloxane 11.5 Liquid paraffin 4.5 Polyoxyethylene-modified dimethylpolysiloxane 4.0 (aqueous phase) Purified water 50.0 1,3-butylene glycol 4.5 Asene Yak ethanol extract 1.5 Sorbitan sesquioleate 3.0 Preservatives Appropriate amount Flavors Appropriate amount (Preparation method) After heating and stirring the aqueous phase, a well-mixed and pulverized powder portion is added, followed by homomixer treatment. Further, the oil phase mixed by heating is added, and the mixture is treated with a homomixer. Then, a fragrance is added with stirring, and the mixture is cooled to room temperature.

Example 11 Cream (Formulation) Solid paraffin 5.0% by weight Beeswax 10.0 Vaseline 15.0 Liquid paraffin 41.0 Glycerin monostearate 2.0 Polyoxyethylene (20 mol) Sorbitan monolaurate 2 0.0 Soap powder 0.1 Borax 0.2 Acenyacacetone extract 0.05 Acenyacethanol extract 0.05 Sodium bisulfite 0.03 Ethylparaben 0.3 Perfume Appropriate amount Ion exchange water Residue (Production method) Ion exchange water Soap powder and borax are added to the mixture, and the mixture is dissolved by heating and maintained at 70 ° C. (aqueous phase). The other components are mixed, melted by heating and kept at 70 ° C. (oil phase). The oil phase is gradually added to the aqueous phase while stirring to carry out the reaction. After completion of the reaction, the mixture is uniformly emulsified with a homomixer, and cooled to 30 ° C. while stirring well after emulsification.

[0029]

As described above, the tyrosinase inhibitor of the present invention has an excellent tyrosinase activity inhibitory effect, and can be used to lighten pigmentation, spots, freckles, liver spots, etc. after sunburn, It has excellent effects and is also excellent in safety.

Claims (2)

[Claims] [Claim 1] Asenyak (scientific name: Uncaria gambir Rox
A tyrosinase inhibitor comprising the extract of b.). 2. The amount of the extract of Acacia catechu is 0.00
The tyrosinase inhibitor according to claim 1, wherein the amount is from 0.5 to 20.0% by weight.