JP4842550B2 - Tyrosinase activity inhibitor, melanin production inhibitor, and skin whitening external preparation - Google Patents

Description

  TECHNICAL FIELD The present invention relates to a tyrosinase activity inhibitor, a melanin production inhibitor and a whitening skin external preparation using N-hydroxysuccinimide.

  Conventionally, skin external preparations such as emulsions, creams, lotions, packs, cleaning agents, dispersions, ointments, liquids, aerosols, patches, poultices, liniments, and the like have been given a prescribed medicinal effect. Various medicinal ingredients are added for the purpose. For example, whitening agents such as arbutin and ascorbic acids (for example, VC-PMg) have been added in order to prevent or improve skin darkening caused by sunburn, stains caused by pigmentation, buckwheat etc. (for example, patents) Documents 1 and 2, Non-patent documents 1 and 2). However, in these whitening agents, the stability of the external preparation for skin may be lowered, or the desired drug efficacy may not be obtained due to deterioration in the preparation, and improvement thereof has been desired.

JP-A-44-031237 Japanese Patent Laid-Open No. 60-016906 FRAGANCE JOURNAL Temporary sensitization No. 14 (1995), p. 151-155, "Whitening action of L-ascorbyl magnesium phosphate" FRAGANCE JOURNAL Temporary sensitization No. 14 (1995), p. 127-132, “Melanin production suppression mechanism and whitening effect of arbutin”

  This invention makes it a subject to provide the novel tyrosinase activity inhibitor and melanin production inhibitor which can be mix | blended stably and was excellent in the whitening effect. Moreover, this invention makes it a subject to provide the skin external preparation which was excellent in stability and excellent in the whitening effect.

  As a result of intensive studies, the present inventor has obtained the knowledge that N-hydroxysuccinimide directly inhibits the activity of tyrosinase, and has completed the present invention based on this knowledge. N-hydroxysuccinimide has a chelating effect on copper, and this chelating effect deprives copper from tyrosinase. As a result, it is considered that the activity of tyrosinase is inhibited. For example, even when melanocytes are activated by sunlight irradiation or the like, the activity of tyrosinase is inhibited in the presence of N-hydroxysuccinimide, so that melanin production is suppressed, resulting in excellent whitening. It is thought that there is an effect.

  That is, this invention provides the tyrosinase activity inhibitor and melanin production inhibitor which use N-hydroxysuccinimide as an active ingredient in order to solve the said subject. Moreover, this invention provides the skin external preparation for whitening which contains N-hydroxysuccinimide as a tyrosinase activity inhibitor or a melanin production inhibitor. Further, from another viewpoint, the present invention suppresses tyrosinase activity by applying N-hydroxysuccinimide, and suppresses melanin production in skin cells by applying N-hydroxysuccinimide. A method is provided.

  According to the present invention, an excellent tyrosinase activity inhibitor and melanin production inhibitor can be provided. Moreover, the skin external preparation containing this exhibits a high inhibitory effect with respect to pigmentation, and is effective in preventing and improving skin darkening due to skin dullness, sunburn, etc., spots and freckles. Therefore, an external preparation for skin containing N-hydroxysuccinimide as a tyrosinase activity inhibitor and / or a melanin production inhibitor can be advantageously used in cosmetics and pharmaceuticals for the purpose of whitening and / or skin.

Hereinafter, the present invention will be described in detail.
The present invention relates to a tyrosinase activity inhibitor containing N-hydroxysuccinimide as an active ingredient and a melanin production inhibitor. In the present invention, a hydrate of N-hydroxysuccinimide may be used. N-hydroxysuccinimide has a chelating effect on copper as described above. For example, an aqueous solution of copper chloride (CuCl ₂ ) exhibits an absorption maximum in the vicinity of a wavelength of 800 nm. However, when N-hydroxysuccinimide is added to the aqueous copper chloride solution, N-hydroxysuccinimide and copper form a chelate complex. As a result, the wavelength of the absorption maximum shifts. The formation of the chelate complex can also be confirmed by tracking the pH change of the solution. In the present invention, the activity of tyrosinase is directly inhibited by using the chelating effect of N-hydroxysuccinimide on copper to suppress melanin production.

  N-hydroxysuccinimide can be blended in various forms of external skin preparations in accordance with conventional methods as a whitening and skin-beautifying component by tyrosinase activity inhibitory action and / or melanin production inhibitory action. The content of N-hydroxysuccinimide as a tyrosinase activity inhibitor and / or a melanin production inhibitor in the external preparation for skin of the present invention is preferably 0.0001 to 5% by mass (hereinafter simply referred to as “%”). Yes, more preferably 0.001 to 1%. If it is in this range, N-hydroxysuccinimide can be blended stably, it is excellent in tyrosinase activity inhibitory action and melanin production inhibitory action, and can exhibit high whitening and / or skin-beautifying effects.

  Examples of the external preparation for skin are not particularly limited, and examples thereof include emulsions, creams, lotions, packs, cleaning agents, makeup cosmetics, dispersions, ointments, liquids, aerosols, patches, poultices, liniments and the like. Any form of cosmetics may be used, such as external medicines.

  In addition to the above essential components, the external preparation for skin of the present invention includes various components that are usually used in cosmetics, quasi-drugs, external medicines, etc., that is, water, alcohol, oils, surfactants, thickeners. Agents, powders, chelating agents, pH adjusting agents, various medicinal agents, extracts derived from animals, plants and microorganisms, fragrances, and the like can be added as appropriate within the range not impairing the effects of the present invention. In addition, N-hydroxysuccinimide and a conventionally known whitening agent, antioxidant, cell activator, anti-inflammatory agent, anti-ultraviolet agent, and other medicinal ingredients are used as a skin external preparation. The effects of these medicinal agents are added to the effects of succinimide, and the effects of the present invention can be further enhanced.

The present invention will be described more specifically with reference to the following examples. However, the scope of the present invention is not limited to the following examples.
[Example 1: Tyrosinase activity inhibitory effect]
Mushroom tyrosinase was dissolved in 0.1 mol / L phosphate buffer so as to be 100 units / mL to prepare a tyrosinase solution. L-Dopa was similarly dissolved in 0.1 mol / L phosphate buffer to prepare a 0.1% L-Dopa solution. N-hydroxysuccinimide was added to a 0.1% L-Dopa solution kept at 37 ° C. at various concentrations (0 to 100 μg / mL), and then the tyrosinase solution was added. The average reaction rate at 37 ° C. at each concentration was calculated, and the concentration (IC ₅₀ ) capable of 50% reaction inhibition was calculated from the obtained results, and the direct inhibitory effect of tyrosinase was evaluated. Moreover, IC ₅₀ was calculated by the same method for arbutin and VC-PMg (ascorbic acid magnesium phosphate) known as tyrosinase activity inhibitors, and the results were compared. The calculated IC ₅₀ is shown in Table 1 below.

  From the results shown in Table 1, it can be understood that N-hydroxysuccinimide directly has a high inhibitory effect on tyrosinase and is an excellent tyrosinase activity inhibitor.

[Example 2: Inhibitory effect on melanin production]
A suitable amount of medium is collected in two 6-well petri dishes, seeded with mouse-derived B16 melanoma cells, and allowed to stand at 37 ° C. in a carbon dioxide concentration of 5 v / v%. The next day, the sample preparation solution is added and mixed so that each sample has an appropriate final concentration. On the fifth day of culture, the medium is changed and the sample preparation solution is added again. On the next day, the medium was removed, the cells were washed with a phosphate buffer in one petri dish, and then collected, and the degree of whitening of the cultured B16 melanoma cells was visually evaluated according to the following criteria.
(Criteria)
++: Extremely white relative to the control.
+: Clearly white relative to the control.
±: Slightly white with respect to the control.
-: The same black color as the control.

  For the remaining one petri dish, cells were fixed with formalin, and then a 1% crystal violet solution was added and stained. The number of viable cells (A) and the number of control cells (B) for each analyte concentration were measured with a monocellator, and the cell viability was calculated by the ratio of (A) / (B)%. In the visual evaluation, the minimum concentration (minimum effective concentration) at which a determination of ± or more was obtained and the concentration (cytotoxic concentration) at which the cell growth rate was less than 70% were determined. For arbutin and VC-PMg known as melanin production inhibitors, the minimum effective concentration and cell growth rate were calculated by the same method, and the results were compared. It is desirable for use to be effective at lower concentrations (ie, have a lower minimum effective concentration) and have higher cytotoxic concentrations. It evaluated from this viewpoint.

  From the results shown in Table 2, since N-hydroxysuccinimide has a high melanin production inhibitory effect and low cytotoxicity, it is an excellent melanin production inhibitor compared to conventionally known melanin production inhibitors. I understand that.

[Example 3: lotion]
(Ingredient) (%)
(1) Glycerin 5.0
(2) 1,3-butylene glycol 6.5
(3) Polyoxyethylene (20E.O.) sorbitan monolaurate ester 1.2
(4) Ethyl alcohol 12.0
(5) Ascorbyl palmitate * 1 0.1
(6) Sodium ascorbate * 2 0.5
(7) Ascorbic acid glucoside * 3 2.0
(8) Lactic acid 0.05
(9) Sodium lactate 0.1
(10) N-hydroxysuccinimide 0.005
(11) Collagen 1.0
(12) Preservative Appropriate amount (13) Fragrance Appropriate amount (14) Purified water Remaining amount * 1 Wako Pure Chemicals * 2 Sigma Co. * 3 Hayashibara Biochemical Research Institute

(Manufacturing method)
A. Components (3) to (5) and (12) and (13) are mixed and dissolved.
B. Components (1), (2), (6) to (11) and (14) are mixed and dissolved.
C. A and B were mixed and uniformed to obtain a skin lotion.

[Example 4: Latex]
(Ingredient) (%)
(1) Polyoxyethylene (10E.O.) sorbitan monostearate 1.0
(2) Polyoxyethylene (60E.O.) sorbit tetraoleate 0.5
(3) Glyceryl monostearate 1.0
(4) Stearic acid 0.5
(5) Behenyl alcohol 0.5
(6) Squalane 8.0
(7) Retinol palmitate * 1 0.0002
(8) Ethyl alcohol 5.0
(9) Daylily extract * 2 0.01
(10) Dipotassium glycyrrhizinate * 3 0.1
(11) N-hydroxysuccinimide 0.1
(12) Purified water remaining amount (13) Preservative 0.1
(14) Carboxyvinyl polymer 0.2
(15) Sodium hydroxide 0.1
(16) Hyaluronic acid 0.1
(17) Zinc oxide Appropriate amount (18) Fragrance Appropriate amount * 1 Made by Nippon Roche Co., Ltd. * 2

(Manufacturing method)
A. Ingredients (12) to (15) are heated and mixed and maintained at 70 ° C.
B. Ingredients (1) to (9) are heated and mixed and maintained at 70 ° C.
C. A is added to B, mixed and uniformly emulsified.
D. After cooling C, (10), (11) and (16) to (18) were added and mixed uniformly to obtain an emulsion.

[Example 5: Cream]
(Ingredient) (%)
(1) Cetostearyl alcohol 3.0
(2) Glyceryl monostearate 2.0
(3) Polyoxyethylene monooleate (20E.O.) sorbitan 1.0
(4) Sorbitan monostearate 1.0
(5) N-stearoyl-N-methyltaurine sodium 0.5
(6) Vaseline 5.0
(7) Dimethylpolysiloxane (100 mm ² / s) 3.0
(8) Glyceryl tri-2-ethylhexanoate 20.0
(9) dl-α-tocopherol * 1 1.0
(10) Ellagic acid * 2 0.05
(11) Lactic acid (50% aqueous solution) 1.0
(12) Dipropylene glycol 10.0
(13) Arbutin * 3 3.0
(14) Sodium citrate 0.5
(15) N-hydroxysuccinimide 0.5
(16) L-ascorbic acid phosphate ester
Magnesium salt * 4 0.1
(17) Titanium oxide 0.1
(18) Perfume appropriate amount (19) Disodium edetate 0.03
(20) Preservative Appropriate amount (21) Purified water remaining * 1 Eisai * 2 Sigma * 3 Wako Pure Chemical * 4 Wako Pure Chemical

(Manufacturing method)
A. Ingredients (1) to (10) are heated and mixed and maintained at 70 ° C.
B. Ingredients (11) to (16) and (19) to (21) are heated and mixed and maintained at 70 ° C.
C. A is added to B, mixed and uniformly emulsified.
D. After cooling C, (17) and (18) were added and mixed uniformly to obtain a cream.

[Example 6: Pack]
(Ingredient) (%)
(1) Polyvinyl alcohol 15.0
(2) Silicic anhydride 0.5
(3) Polyethylene glycol 0.5
(4) Polyoxypropylene (10 PO) methyl glucoside 0.5
(5) Glycerin 5.0
(6) Purified water remaining amount (7) Ethyl alcohol 10.0
(8) Preservative appropriate amount (9) N-hydroxysuccinimide 0.05
(10) Perfume appropriate amount

(Manufacturing method)
A. Components (1) to (6) are mixed and heated to 70 ° C. to dissolve.
B. Components (7) to (8) are mixed and dissolved.
C. B was added to the previous A, mixed, and then cooled to uniformly disperse (9) and (10) to obtain a pack.

[Example 7: Liquid foundation]
(Ingredient) (%)
(1) Dipentaerythritol tetra 12-hydroxystearic acid sesquistearic acid hemirosin ester 2.0
(2) Liquid paraffin 5.0
(3) Stearic acid 2.0
(4) Cetanol 1.0
(5) Self-emulsifying glyceryl monostearate 1.0
(6) Paramethoxycinnamic acid-2-ethylhexyl 8.0
(7) N-hydroxysuccinimide 0.01
(8) Preservative appropriate amount (9) Glycerin 5.0
(10) Triethanolamine 1.0
(11) Carboxymethylcellulose 0.2
(12) Bentonite 0.5
(13) Purified water remaining amount (14) Titanium oxide 6.0
(15) Fine particle titanium oxide 2.0
(16) Fine zinc oxide 5.0
(17) Mica 2.0
(18) Talc 4.0
(19) Color pigment 4.0
(20) Perfume appropriate amount

(Manufacturing method)
A. Ingredients (1) to (6) are heated and mixed and dissolved.
B. Ingredients (14) to (19) are added to A, mixed uniformly, and kept at 70 ° C.
C. Ingredients (8) to (13) are uniformly dissolved and kept at 70 ° C.
D. B is added to C and emulsified uniformly.
E. After cooling D, components (7) and (20) were added to obtain a liquid foundation.

[Example 8: Sunscreen emulsion]
(Ingredient) (%)
(1) Polyoxyalkylene-modified organopolysiloxane 1.0
(2) Dimethylpolysiloxane 5.0
(3) Octamethylcyclotetrasiloxane 20.0
(4) Isotridecyl isononanoate 5.0
(5) Paramethoxycinnamic acid-2-ethylhexyl 5.0
(6) Fine particle titanium oxide 10.0
(7) Fine zinc oxide 10.0
(8) Zirconium oxide 5.0
(9) Polystyrene powder 3.0
(10) Trimethylsiloxysilicate 0.5
(11) Preservative appropriate amount (12) Dipropylene glycol 3.0
(13) Ethyl alcohol 10.0
(14) Purified water remaining amount (15) Salt 0.2
(16) N-hydroxysuccinimide 0.2
(17) Appropriate amount of perfume

(Manufacturing method)
A. Components (1) to (10) are mixed and dispersed.
B. Components (11) to (15) are mixed and dissolved.
C. Add B to A and emulsify uniformly.
D. Components (16) and (17) were added to C to obtain a sunscreen emulsion.

  The cosmetics of Examples 3 to 8 containing N-hydroxysuccinimide as a tyrosinase activity inhibitor were all excellent in the whitening effect and did not cause precipitation of components, odor change or discoloration.

  The tyrosinase activity inhibitor and melanin production inhibitor of the present invention are useful as components of various skin external preparations including cosmetics, in particular, skin whitening external preparations.

Claims (2)

A tyrosinase activity inhibitor comprising N-hydroxysuccinimide as an active ingredient. A melanin production inhibitor containing N-hydroxysuccinimide as an active ingredient.