JP5677126B2 - Tyrosinase activity inhibitor and whitening cosmetics - Google Patents

Description

  The present invention relates to a tyrosinase activity inhibitor and a cosmetic for whitening.

  Conventionally, whitening cosmetics are blended with ingredients that suppress the formation of melanin, which is the cause of dark browning of the skin. Such components that suppress the production of melanin include components that inhibit tyrosinase activity due to cytotoxicity, components that suppress tyrosinase activity in melanocytes, components that suppress tyrosinase expression, and dopaquinone produced by tyrosinase activity Ingredients that suppress the pathway leading to melanin by auto-oxidation are known.

  For example, cysteine, glutathione, vitamin C, kojic acid, products of microorganisms belonging to the genus Trichoderma (patent document 1), silk protein alkaline degradation product (patent document 2), milk protein hydrolyzate (patent document 3) , Amino acid derivatives and peptide derivatives of kojic acid (Patent Document 4), fragrance compound group having a tyrosinase activity inhibiting function (Patent Document 5), fragrance compound group having a tyrosinase activity inhibiting function (Patent Document 6, Patent Document 7), various An inhibitory action on tyrosinase activity of plant extracts and the like is known.

  However, these conventional tyrosinase activity inhibitors, such as hydroquinone, have safety problems, are unsatisfactory in practical use, have strong side effects, and cannot be used as cosmetics. It was not satisfactory.

  On the other hand, the saffron extract is known to have a damage recovery action by ultraviolet rays, a hyaluronic acid acid promoting action, and an aquaporin synthesis promoting action (see Patent Documents 8 to 11). The saffron extract has been reported to have a tyrosinase activity promoting action (see Patent Document 12), but the tyrosinase activity inhibitory effect has not yet been reported.

Japanese Patent Laid-Open No. 2-145189 Japanese Patent Publication No.58-017763 JP-A-5-320068 Japanese Patent Laid-Open No. 4-187618 JP 2000-302642 A JP 2001-163719 A JP 2001-240528 A JP-A-11-349468 JP-A-2005-206510 JP 2005-206512 A JP 2005-343882 A Japanese Patent Laid-Open No. 11-79950

  An object of the present invention is to provide a tyrosinase activity inhibitor having an excellent tyrosinase activity inhibitory effect, which is safe and weak in skin irritation, and a whitening cosmetic product containing the same.

  Therefore, as a result of intensive research in view of the above circumstances, the present inventors have found that the glucosidase-treated product of Crocus sativus extract has a remarkably superior tyrosinase activity inhibitory effect compared to the saffron extract itself. The present invention was completed by finding that the skin irritation was low and had an excellent whitening effect.

  That is, the present invention provides a tyrosinase activity inhibitor comprising as an active ingredient a glucosidase-treated product of a Crocus sativus extract. Moreover, this invention provides the cosmetics for whitening which use the glucosidase processed material of the Iridaceae crocus genus saffron (Crocus sativus) extract as an active ingredient.

  The glucosidase-treated product of the saffron extract has an excellent tyrosinase activity inhibitory effect and whitening effect, and has low irritation to the skin, so it is useful as a tyrosinase activity inhibitor and a whitening cosmetic product. Moreover, it is excellent in safety | security, and arbitrary amounts can be mix | blended with cosmetics, without impairing the quality of cosmetics. It is useful as a whitening cosmetic effective in preventing and improving pigmentation, stains, freckles, liver spots, etc. after sunburn. Even if it is not a cosmetic, it is expected to be applied to pharmaceuticals and foods.

  The active ingredient of the tyrosinase activity inhibitor and whitening cosmetic product of the present invention is a glucosidase-treated product of an extract of Crocus sativus.

  Crocus sativus used in the present invention is a plant distributed in the northern hemisphere and gives purple flowers in autumn. Examples of the saffron extract include saffron leaf and flower extracts, but the whole flower (corolla, bud, pistil, stamen) or female core head extract is preferable, and the whole flower extract is more preferable.

  The saffron extract can be obtained by a solvent extraction method, a liquefied gas extraction method such as liquefied carbon dioxide gas or liquefied butane gas, or a supercritical gas extraction method. Solvents used here include water or alcohols such as methanol, ethanol and isopropyl alcohol, polyhydric alcohols such as ethylene glycol, propylene glycol and 1,3-butylene glycol, ketones such as acetone, and esters such as ethyl acetate. , Ethers such as diethyl ether, aromatic compounds such as hexane, pentane and benzene can be selected from one kind alone or a mixture of two or more kinds. Of these, it is preferable to use ethanol and / or ethyl ether.

  In the present invention, a glucosidase-treated product of saffron extract is used. The glucosidase used in the present invention is an enzyme that cleaves sugar chains such as glycoproteins and glycolipids. For example, endo-β-N-acetylglucosaminidase, β-N-acetylhexosaminidase, endo-α-N. -Acetylgalactosaminidase, α-L-fucosidase, α-N-acetylgalactosaminidase, transglucosidase, glycopeptidase, N-glycanase, etc., and plants such as almonds and peanuts, Flavobacterium sp. Alcarigenes sp. , Aspergillus sp. Those originating from microorganisms such as can be used.

  In the present invention, glucosidase is used alone or in combination of two or more. As a method for treating saffron extract with glucosidase, glucosidase is directly added to saffron extract, or glucosidase is used as an immobilized enzyme. Although there is a method of passing a saffron extract through a column packed with, there is no particular limitation.

  The treatment temperature with the glucosidase of the present invention is preferably in the temperature range of 25 to 70 ° C. By setting the treatment temperature to 25 ° C. or higher, it is possible to prevent a long time from being decomposed due to a decrease in enzyme activity. In addition, by setting the treatment temperature to 70 ° C. or lower, it is possible to prevent the enzyme from being severely deactivated and difficult to control the decomposition reaction.

  In the glucosidase treatment, it is preferable to use the saffron (Crocus sativus) extract as it is or after dissolving the saffron (Crocus sativus) extract in water so that the solid content concentration is in the range of 2 to 40% by mass. Yield reduction can be prevented by setting the solid content concentration to 2% by mass or more. Moreover, by setting it as 40 mass% or less, the viscosity of a solution rises, stirring becomes difficult, and problems, such as becoming difficult to fully melt | dissolve, can be prevented.

  The saffron extract glucosidase-treated product obtained by the method of the present invention can be powdered as it is, in the form of a liquid concentrated by a method such as vacuum concentration, or by a method such as spray drying or freeze drying. It can be used in the form of a powder obtained by extraction or extraction with an organic solvent such as hexane, ethyl ether, ethyl acetate, methanol, ethanol, etc. to remove untreated substances and then the solvent.

  In the tyrosinase activity inhibitor of the present invention, a glucosidase-treated product of the saffron extract may be used alone, or may be used in combination with known tyrosinase activity inhibitors such as arbutin, kojic acid, ascorbic acid and the like. it can. It can also be used in combination with other active ingredients such as vitamins, antioxidants, anti-inflammatory agents, ultraviolet absorbers, cooling agents, and the like. Others, ethanol, isopropanol, butyl alcohol, benzyl alcohol, isostearyl alcohol, octyldodecanol, hexyl decanol and other alcohols, ethylene glycol, diethylene glycol, triethylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, polypropylene glycol, glycerin, Cosmetics such as diglycerin, polyglycerin, isoprene glycol, polyhydric alcohols such as 1,3-butylene glycol, triglycerides such as avocado oil, olive oil, sesame oil, wheat germ oil, ester oils, hydrocarbons, surfactants, etc. It can be used in combination with commonly used components.

  The tyrosinase activity inhibitor of the present invention is preferably 50 to 100% by mass, more preferably 65 to 100% by mass, and particularly preferably 75 to 100% by mass, in terms of solid content, of the glucosidase-treated product of the saffron extract based on the total amount of the composition. Contains by mass%. If it is in the said range, the outstanding inhibitory effect of tyrosinase activity will be acquired and it is preferable.

  The tyrosinase activity inhibitor of the present invention can be in various forms such as an aqueous composition, an oily composition, an emulsified composition, a liquid composition, a solid composition, a powder composition, and a paste composition.

  The tyrosinase activity inhibitor of the present invention is a whitening foundation, lotion, lotion, cream, gel, pack, shampoo, rinse, body soap, face wash, and other cosmetics, soaps, detergents and other skin. It can be suitably used by adding to cosmetics such as external preparations, perfumes and fragrances. Moreover, even if it is other than cosmetics, the application to a pharmaceutical and food-drinks can also be expected.

The whitening cosmetic product of the present invention is preferably 0.001 to 20% by mass, more preferably 0.005 to 5% by weight in terms of solid content of the glucosidase-treated saffron extract based on the total amount of the cosmetic product. 10 mass%, More preferably, it contains 0.05-5 mass%, Most preferably, it contains 0.01-1 mass%. If it is in the said range, the outstanding tyrosinase activity inhibitory effect and the whitening effect will be acquired, and skin irritation is also low, and it is preferable.
In the cosmetic for whitening of the present invention, the components described as components that can be blended in the tyrosinase activity inhibitor can be blended.

Hereinafter, although this invention is demonstrated based on a test example and an Example, this invention is not limited to these. In addition, a compounding quantity is the mass%.
In addition, Example 2 in Table 1 and Examples 6 and 7 in Table 2 are reference examples and are not included in the scope of claims.

[Extraction method]
After saffron (Crocus sativus) flowers were immersed in 95% ethanol, insoluble matter was filtered, and then ethanol was removed to obtain a saffron (flower) extract (yield 5.1%).
Similarly, pistils were separated from saffron (Crocus sativus) flowers, immersed in 95% ethanol, insoluble matter was filtered, ethanol was then removed, and a saffron (pistil) extract was obtained (yield 3.5%). .

[Glucosidase treatment method]
A saffron (flower) extract (1 g) and distilled water (9 g) are mixed and heated in a warm bath (70 ° C.) to dissolve. After complete dissolution, cool to 36 ° C. and add enzyme β-glucosidase (5 mg, 185 U). Thereafter, the ink is incubated at 37 ° C. for 1 hour. After the reaction, the solution is extracted with ethyl ether. The extract was washed twice with water, and the ether solution was dried with magnesium sulfate to obtain a saffron (flower) extract glucosidase-treated product.
Similarly, saffron (pistil) extract (1 g) and distilled water (9 g) are mixed and heated in a warm bath (70 ° C.) to dissolve. After complete dissolution, cool to 36 ° C. and add enzyme β-glucosidase (5 mg, 185 U). Thereafter, the ink is incubated at 37 ° C. for 1 hour. After the reaction, the solution is extracted with ethyl ether. The extract was washed twice with water, and the ether solution was dried over magnesium sulfate to obtain a saffron (pistil) extract-treated glucosidase product.

[Tyrosinase activity inhibition test]
(Test method)
A saffron (flower) extract, a saffron (pistil) extract, a saffron (flower) extract glucosidase-treated product and a saffron (pistil) extract glucosidase-treated product were each dissolved in hexane to prepare 0.5 mass% solutions. Ethanol was further added thereto and diluted 10 times to prepare a 0.05% by mass sample solution.
10 μl of this sample solution and 85 μl of 0.1 M phosphate buffer (pH 7), 5 μl of 5000 U / ml tyrosinase (derived from mushrooms: manufactured by SIGMA ALDRICH) were mixed with stirring and incubated at 37 ° C. for 5 minutes.
After 5 minutes, the absorbance (A) at 475 nm was measured with a spectrophotometer (Multi-Spectrophotometer Vient XS / Dainippon Sumitomo Pharma) to obtain a tyrosinase activity inhibition rate (%) ID ₅₀ .
On the other hand, the same operation was performed with no sample solution added, and the obtained absorbance (B) was measured.

(ID ₅₀ calculation formula)
Tyrosinase activity inhibition rate (%) = ((B) − (A)) / (B) × 100
(However, A: Absorbance of sample, B: Absorbance of blank)

Similarly, each sample shown in Table 1 was tested. A graph was prepared by plotting the concentration on the horizontal axis and the tyrosinase inhibition rate on the vertical axis. From this graph, the concentration at which tyrosinase activity was inhibited by 50% (hereinafter referred to as ID ₅₀ , unit μg / ml) was determined. The results are shown in Table 1. In addition, the saffron extract used for the comparative example had an ID ₅₀ value exceeding 1000, and no tyrosinase activity inhibitory action was observed.

  As is apparent from Table 1, the glucosidase-treated product of the saffron extract had a particularly remarkable tyrosinase activity inhibitory action compared to the saffron extract.

Examples 1-7 and Comparative Examples 1-2 (skin lotion)
A skin lotion containing a saffron extract treated with glucosidase was prepared by the following method, and the whitening effect and skin irritation were evaluated. The results of the prepared formulation, whitening effect and skin irritation test are shown in Table 2.

<Preparation method of skin lotion>
The aqueous phase and the alcohol phase are each uniformly dissolved, the aqueous phase and the alcohol phase are mixed and dispersed by stirring and solubilizing, and then filled into a container. In use, the contents are uniformly shaken and dispersed.

<Test method>
The skin of the forearm bent side of 20 subjects exposed to sunlight for 3 hours (1.5 hours per day for 2 days) was used as the test site, and the left forearm bent side skin from the day exposed to sunlight. The sample was applied to the right forearm flexion side skin on a base not containing the sample once every morning and evening for 13 weeks. A whitening effect before and after continuous application and skin irritation during the application period were evaluated by a specialist judge.
In addition, about the evaluation result of the whitening effect, it showed as a test subject's number of persons by whom the whitening effect was confirmed in the sample application part from the base application part. Skin irritation was shown as the number of subjects who felt abnormalities (erythema, edema, crust) or tingling on the skin of the application area.

  As is clear from the results shown in Table 2, Examples 1 to 7 containing the saffron extract treated with glucosidase were confirmed to have excellent whitening effects and low skin irritation. On the other hand, the whitening effect was hardly confirmed in the comparative example using the saffron extract instead of the glucosidase-treated product of the saffron extract.

  Examples of prescriptions for tyrosinase activity inhibitors and cosmetics using saffron extract treated with glucosidase are shown below. In addition, the tyrosinase activity inhibitor and cosmetics shown here are only examples, and the fragrance is not limited to these, and an attractive fragrance rich in variety can be expressed.

Formulation Example 1 Fragrance Oil (Floral Formula)

Formulation Example 2 Skin Lotion Component Content (%)
1. Ethanol 8
2. Polyoxyethylene sorbitan monolaurate (20E.O.) 0.4
3. Methylparaben 0.1
4.1,3-Butylene glycol 5
5. N-methyl-L-serine 0.1
6). Fire thorn extract 0.1
7). Saffron (flower) extract treated with glucosidase 0.1
8). Fragrance 0.01
9. Purified water balance

Formulation Example 3 Skin Cream Component Content (%)
1. Glycerol monostearate 2
2. Beeswax 1
3. Sorbitan monostearate 1
4). Vaseline 4
5. Liquid paraffin 12
6). N-stearoyl-L-glutamate sodium 1
7). Xanthan gum 0.2
8). Cam Come Extract 0.1
9. Methylparaben 0.1
10. Saffron (pistil) glucosidase processed product 0.1
11. Purified water balance

Formulation Example 4 Day Essence Component Content (%)
1. Stearic acid 0.3
2. Lipophilic glyceryl monostearate 2
3. Tri-2-ethylhexanoic acid glycerin 5
4). Parasol MCX 5
5. Parasol 1789 5
6). N-stearoyl-sodium L-glutamate 0.5
7). Glycerin 5
8). Yukinoshita extract 0.1
9. Xanthan gum 0.15
10. Titanium dioxide 5
11. Saffron (flower) extract treated with glucosidase 0.5
12 Purified water balance

  The obtained tyrosinase activity inhibitor and cosmetics had a good fragrance balance and had excellent odor characteristics similar to those of cosmetic products using conventional floral fragrances.

  The tyrosinase activity inhibitor and whitening cosmetic product of the present invention are excellent in safety and can be blended in an arbitrary amount in the cosmetic product without impairing the quality of the cosmetic product. It is useful as a whitening cosmetic effective in preventing and improving pigmentation, stains, freckles, liver spots, etc. after sunburn. Even if it is not a cosmetic, it is expected to be applied to pharmaceuticals and foods.

Claims (2)

A tyrosinase activity inhibitor comprising, as an active ingredient, a glucosidase-treated product of a Crocus sativus flower extract (excluding an extract of pistil only) . A cosmetic for whitening comprising a glucosidase treated product of Crocus sativus flower extract (excluding pistil extract) as an active ingredient.