JP2006001866A - Melanin production inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To search a substance for melanin production inhibition, which has tyrosinase inhibitory activity and is useful as an active ingredient for a bleaching ingredient, and a melanin production inhibitor using the same. <P>SOLUTION: It is reported that, when the amount of nucleic acids in cells is increased, the amount of melanin produced is reduced. As a result of examination by paying attention to a glycolytic product giving a final product to become lactic acid as one method for increasing lactic acid in cells, the glycolytic product is confirmed to remarkably inhibit melanin production of B16 melanoma cell, the glycolytic product is found to become a melanin production inhibitor having inhibitory action on melanin production and promotion effect on percutaneous absorption by use together with a phospholipid, thus this invention is completed. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a melanin production inhibitor that can be expected to have a whitening effect.

  Conventionally, it is known that skin spots and buckwheat are caused by the production of melanin in tissues by sunburn or the like. Melanin is also known to be formed through several steps after tyrosine is oxidized enzymatically by tyrosinase. Therefore, by inhibiting the activity of tyrosinase, the production of melanin can be suppressed, and the occurrence of skin spots, buckwheat and the like can be suppressed. Arbutin, kojic acid, casein hydrolyzate, and the like are known as substances that inhibit the activity of tyrosinase, and are used as active ingredients of whitening agents. Moreover, the present condition is that search is carried out in search of the substance which has tyrosinase activity inhibition and can be used as an active ingredient of a whitening agent.

  Known novel whitening agents include isonitrile compounds produced by filamentous fungi, phosphorylated polysaccharides produced by lactic acid bacteria, naphthol glycosides, and whitening external preparation compositions containing the same. (See Patent Documents 1, 2, 3, 4, and 5)

  As a report that has recently attracted attention, it has been reported that the expression level of tyrosinase, which is a melanin-producing rate-determining enzyme, is suppressed by lactic acid, and as a result, the amount of melanin decreases. (See Non-Patent Document 1)

In addition, it has been reported in the literature that the amount of melanin produced decreases when intracellular lactic acid increases. (See Non-Patent Document 2)
JP-A-9-249524 JP 7-236491 A JP-A-7-242525 JP-A-7-242687 JP 2003-238585 A J Invest Dermatol. 1993 Feb; 100 (2 Suppl): 150S-155S Tyrosinase gene transcription and its control by melanogenic inhibitors.Ando S, Ando O, Suemoto Y, Mishima Y. J Biol Chem. 2002 Apr 26; 277 (17): 14821-8. Epub 2002 Jan 25. Abnormal acidification of melanoma cells induces tyrosinase retention in the early secretory pathway.Halaban R, Patton RS, Cheng E, Svedine S, Trombetta ES , Wahl ML, Ariyan S, Hebert DN.

  A major factor in stain formation is the promotion of melanin production in melanocytes. Accordingly, the present invention searches for a melanin production inhibitor that has tyrosinase activity inhibition and can be used as an active ingredient of a whitening agent, and provides a melanin production inhibitor using the substance.

  It has been reported that when intracellular lactic acid increases, the amount of melanin produced decreases. The inhibitory action of melanin production by lactic acid is known from Non-Patent Documents 1 and 2 above, but a method for increasing intracellular lactic acid was investigated by searching for a sample that acts in a more upstream process. As a method of increasing intracellular lactic acid, as a result of investigating glycolytic products in which the final product is lactic acid, glycolytic products significantly suppress melanin production in B16 melanoma cells. As a result, it has been found that by using phospholipid in combination, it can be a melanin production inhibitor having a melanin production inhibitory effect and a transdermal absorption promoting effect, and the present invention has been completed.

That is, the present invention is a melanin production inhibitor containing a glycolytic product excluding lactic acid and ethanol and a phospholipid. When the melanin production inhibitor further contains a whitening agent, the effect is improved. Moreover, the melanin production inhibitor of this invention can be a skin external preparation.
According to the present invention, the enhancement of transdermal absorption of glycolytic products can be enhanced, and by supplying more glycolytic products into cells, intracellular lactic acid can be increased, and a melanin production inhibitor As a result, inhibition of melanin production can be efficiently expressed in vivo. Furthermore, the combined effect of melanin production suppression is exhibited by using together with known whitening agents such as vitamin C.

  The melanin production inhibitor using the glycolytic product and phospholipid in combination resulted in remarkable effects both in the melanin production inhibitory effect and the percutaneous absorption promoting effect.

  The glycolytic product used in the present invention is a compound that is metabolized in the glycolytic pathway that is a metabolic pathway of sugar. Glycolysis pathway means a metabolic pathway that is decomposed from glucose metabolic system that is common to almost all living organisms to pyruvic acid, ethanol, or lactic acid. Among them, glucose metabolic system is preferable. The reaction takes place in the cytoplasm.

  Specific examples of the glycolytic product include glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-bisphosphate, dihydroxyacetophosphate, glyceraldehyde 3-phosphate, 1 , 3-bisphosphoglyceric acid, 3-phosphoglyceric acid, 2-phosphoglyceric acid, phosphoenolpyruvate, enolpyruvic acid, pyruvic acid. Lactic acid and ethanol are excluded.

  The blending amount of the glycolytic product is preferably 0.001 to 2% by mass in the total amount of the melanin production inhibitor.

  The phospholipid used in the present invention is not particularly limited to the type thereof, and is not limited to the fatty acid structure regardless of saturated fatty acid / unsaturated fatty acid.

  As the phospholipid, lecithin, lysolecithin obtained from soybean, egg yolk, and the like, or derivatives thereof such as hydrogenated products and hydroxides can be used. Further, phosphatidylcholine, phosphatidic acid, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylglycerol, sphingomyelin and the like can be used.

The blending amount of the phospholipid is preferably 0.01 to 20% by mass, more preferably 0.05 to 10% by mass in the total amount of the melanin production inhibitor.
In the present invention, by using a whitening agent together with a glycolytic product and a phospholipid, a redundant melanin production inhibitory action is recognized.

Suitable whitening agents include L-ascorbic acid, sodium L-ascorbate, sodium ascorbate phosphate, magnesium ascorbate phosphate, sodium ascorbate sulfate, glyceryl ascorbate (polyglyceryl) ethers, glyceryl ascorbate (Polyglyceryl) esters, ascorbic acid glucosides (glycosides), ascorbic acid alkyl esters, ascorbic acid alkyl ethers and other vitamin C and derivatives thereof, kojic acid, arbutin, placenta extract, sulfur and the like.
In addition, plant extracts such as oil-soluble licorice extract, mulberry extract, peony extract, pearl millet extract, bitumen extract, maronier bark extract and chamomile extract, and linoleic acid, linolenic acid, lactic acid, tranexamic acid and the like are also preferred.

  The blending amount of the whitening agent is preferably 0.001 to 5% by mass, more preferably 0.01 to 2% by mass, based on the total amount of the melanin production inhibitor.

  The melanin production inhibitor of the present invention can be blended with other ingredients to be blended with an external preparation for skin such as cosmetics and quasi-drugs as long as the effects of the present invention are not impaired.

  For example, hardened oils derived from animal and vegetable oils, waxes derived from nature, hydrocarbon oil phase components, oil phase components derived from animals and plants, silicone oil phase components, higher alcohols, thickeners, UV absorbers, powders, Pigments, polyhydric alcohols, sugars, polymer compounds, physiologically active ingredients, solvents, antioxidants, fragrances, preservatives, and the like can be blended.

  The melanin production inhibitor of the present invention is preferably applicable to cosmetics and the like as a skin external preparation. Examples of usage forms of the external preparation for skin include ointments, creams, lotions, patches and the like. The base of the external preparation may be a known external base and is not particularly limited.

  The melanin production inhibitor containing a glycolytic product and a phospholipid according to the present invention is excellent in intracellular uptake efficiency, particularly excellent in skin permeability, and effective in melanocytes localized in the epidermal basal layer and peripheral melanocytes. Promotes the suppression of melanin production.

The present invention will be described in detail, but the technical scope of the present invention is not limited to the examples.
(Test Example) Evaluation method for evaluating the effect of B16 melanoma cells on total melanin production B16 mouse melanoma strain F0 (B16F0) was seeded in a 6-well plate at a cell density of 1.0 × 10 ⁴ cells / well.
2. After culturing in 5% FBS-containing DMEM for 24 hours, the culture medium was replaced with 5% FBS-containing DMEM containing a sample of a predetermined concentration.
3. After further culturing for 6 days, the cells were detached by trypsin treatment to prepare a cell pellet. At this time, the color tone of the cell pellet was scored by visual judgment (5-level score: 1 white-5 black).
4). The amount of melanin produced was measured by an alkali solubilization method. The cell pellet was washed with 5% trichloroacetic acid, ethanol / diethyl ether solution (3: 1) and diethyl ether, and 1N sodium hydroxide was added and dissolved by heating (100 ° C, 5 minutes). The absorbance at 430 nm was measured. The amount of melanin was calculated from a calibration curve prepared using synthetic melanin (Sigma) as a standard product. The amount of melanin per unit protein was calculated by dividing the amount of melanin by the amount of protein in all cells.

Test results

Tables 1 to 11 show the results of suppression of melanin production by glycolytic product alone or by combining glycolytic product and lecithin.
Means in the table (Conc .: concentration, Mean: mean, p (t-test): significant difference test, SD: standard deviation).

  From the results shown in Tables 1 to 11, the effect of B16 melanoma cells on total melanin production was evaluated by comparing glycan products alone with glycan products and melanin production inhibitors combined with glycolytic products and lecithin compared to controls. The result was a production inhibitory effect. In addition, the melanin production inhibitor using the glycolytic product and lecithin in combination with the glycolytic product alone was more effective in both the melanin production inhibitory effect and the transdermal absorption promoting effect.

Next, Table 12 shows application examples of the external preparation for skin of the present invention. In addition, a compounding quantity is the mass%.
An external preparation for skin (beauty liquid) was prepared according to the following formulation. In the preparation method, the following components were stirred and mixed at room temperature to prepare an external preparation for skin (beauty liquid).

Next, a rough skin improvement effect test was performed on each sample according to the following method.
(1) Skin roughness improvement effect test

    Set the number of samples to be irradiated by 1 x 1 cm on the forearm of 10 panelists as test subjects (in the case of this use test, a total of 3 samples including blank, glycolytic product, glycolytic product + lecithin) And 0.05 ml of each sample was uniformly applied to each location. Thereafter, a Toshiba UV-B lamp (SE-30E) was set as an artificial ultraviolet light source so as to have an intensity of 0.7 mW / ?, and each part coated with the sample was irradiated for 3 minutes. Thus, 24 hours after irradiation with ultraviolet rays, the degree of inflammation at the irradiated site was observed and evaluated with a 60-fold contact-type skin enlargement video. The results are shown in Table 13.

  From the results shown in Table 13, when the test was actually used in a panel, compared with the blank and lecithin-free glycolytic products, the topical skin preparation (beauty liquid) containing glycolytic products + lecithin was not photoaging. Progression was suppressed, and a remarkable melanin production inhibitory effect was observed.

Other skin external preparation application examples are shown below. All of these external preparations for skin enhance the transdermal absorbability of glycolytic products to the skin, effectively induce the suppression of melanin production in the subepidermal region by external application, and have the effect of suppressing the progress of photoaging. It was confirmed that there was.


[cream]
Ingredient Amount (% by mass)
Stearic acid 5.0
Squalane 2.0
Liquid paraffin 3.0
Glyceryl tri-2-ethylhexanoate 10.0
Glucose-6-phosphate 1.5
Lecithin 5.0
Arbutin 1.0
Glycerol monooleate 2.0
Polyoxyethylene (20 mol) 2.5
Sorbitan monooleate 1,3-butylene glycol 5.0
Glycerin 2.0
Vitamin E 0.05
Preservative Appropriate amount of perfume Appropriate amount of purified water Remaining amount [Essence]
Ingredient Amount (% by mass)
Glycerin 3.0
Dipropylene glycol 10.0
Ethanol 15.0
Carboxyvinyl polymer 0.2
Sodium hyaluronate 0.5
Potassium hydroxide 0.1
Polyoxyethylene (20 mol) 1.0
Sorbitan monooleate polyoxyethylene (20 mol) 1.0
Octyldodecyl ether
Olive oil 0.2
Glucose-6-phosphate 0.1
Lecithin 1.0
Ascorbic acid phosphate magnesium 0.1
Preservative Appropriate amount EDTA / 3Na Appropriate perfume Appropriate amount Purified water remaining

The melanin production inhibitor using the glycolytic product and phospholipid in combination resulted in remarkable effects both in the melanin production inhibitory effect and the percutaneous absorption promoting effect. This melanin production inhibitor can be used to increase intracellular lactic acid, and can lead to the development of an external preparation for skin as a preparation capable of efficiently expressing melanin production inhibition in vivo.

Claims (3)

A melanin production inhibitor containing a glycolytic product excluding lactic acid and ethanol and a phospholipid. Furthermore, the melanin production inhibitor of Claim 1 containing the whitening agent. The melanin production inhibitor of Claim 1 or 2 which is a skin external preparation.