JP4148862B2 - Tyrosinase activity inhibitor - Google Patents

Description

  The present invention relates to a tyrosinase activity inhibitor having an excellent effect, a skin external preparation containing such a tyrosinase activity inhibitor as a whitening active ingredient, and a food applied as a food browning prevention active ingredient.

  Tyrosinase is an oxidase that uses various catechol derivatives as substrates. Most of them have monophenol monooxygenase activity, and mono-phenol is used as a substrate to produce ortho-diphenol, which is further oxidized to ortho-quinones. This oxidase is widely distributed in nature, and is present in pigment cells of plants and animals such as mushrooms, potatoes and apples. Oxidases in animals exhibit high activity especially against tyrosine and dopa, and are deeply involved in melanin biosynthesis. Melanin is produced from tyrosine in the process of L-dopa, dopaquinone, dopachrome, 5,6 dihydroxyindole, indole-5,6-quinone and melanin, and tyrosinase is involved in the oxidation process of tyrosine to L-dopa and dopaquinone. ing. Melanin is a pigment that determines the color of hair and skin. In the skin, this pigment is produced in response to light and ultraviolet rays in melanocytes existing in the basal layer of the epidermis. Melanin production in the skin plays a role in protecting humans from the adverse effects of ultraviolet rays and the like. However, when there is a stimulus such as exposure to a large amount of light or ultraviolet rays, the melanin generating function is locally maintained, and the skin of that part is darkened. As a result, pigmentation will form spots and freckles, which eventually promote skin aging. Melanin pigment is also involved in the phenomenon that the cut surface changes brown when vegetables and fruits such as apples, yams, and lettuce are cut. Fruits and vegetables whose cut surfaces are browned are difficult to store for a long period of time due to their reduced commercial value. Conventionally, tyrosinase activity inhibitors have been used for the purpose of promoting whitening and preventing browning of food. Known examples of tyrosinase activity inhibitors include vitamin C (see Patent Document 1), arbutin, extracts extracted from plants and the like (see Patent Document 2), and sulfites. However, vitamin C is easily oxidized due to its own reducing action and is unstable, and has a storage problem. Arbutin also has a problem in heat resistance stability, which also has a problem in handling. Plant extracts are difficult to identify in chemical composition, and there is a concern about quality variations among lots. Sulfites are recognized as a skin irritation to the human body and have safety problems. On the other hand, aromatic aldehydes used as food fragrances are known to have tyrosinase inhibitory activity (see Non-Patent Document 1), but foods and the like are colored by their own color tone. There's a problem.

Japanese Patent Laid-Open No. 2-45408 JP-A-6-40896 Oil Chemistry, 43 (7), 574-578, 1994

  An object of the present invention is to provide a tyrosinase activity inhibitor that solves the above-described problems, has high safety to the human body, and exhibits high effects.

  In order to solve the above-mentioned problems, various studies were conducted, and high tyrosinase activity was observed in each compound of nerol, vanillin, hydroxycitronellal, methyl heptine carboxylate, benzyl alcohol, methyl naphthyl ketone, rosephenone, and isobornyl acetate. It has been found that the inhibitory action is exhibited, and the present invention has been completed.

  Moreover, the skin external preparation in this invention is characterized by containing the above-mentioned tyrosinase activity inhibitor as a whitening active ingredient.

  Moreover, the foodstuff in this invention applies the above-mentioned tyrosinase activity inhibitor as a browning prevention active ingredient, It is characterized by the above-mentioned.

  The effect of the tyrosinase activity inhibitor of the present invention was measured by the following method.

Evaluation of human epidermal melanocyte tyrosinase activity inhibition Normal human epidermal melanocytes manufactured by Kurabo Industries Co., Ltd. were seeded in a 96-well microplate so as to be 3.0 × 10 ⁴ cells per well. As a seeding medium, Medium154S manufactured by Kurabo Industries Co., Ltd. was used. After 24 hours, the sample solution was adjusted to each concentration with Medium 154S, and further cultured for 48 hours. Next, 75 μL of 1 wt% Triton-X-containing phosphate buffer was exchanged to completely lyse the cells, and 50 μL was used as a crude enzyme solution. To the crude enzyme solution, 50 μL of a 0.05 wt% L-dopa-containing phosphate buffer as a substrate was added and allowed to stand at 37 ° C. for 2 hours. The amount of dopamelanin produced by measuring the absorbance at 405 nm immediately after addition of the substrate and at the end of the reaction with a microplate reader was determined by introducing the difference between the two measured values into the following equation.
405 nm value after reaction-405 nm value after reaction = 5.238 x (amount of dopamelanin produced) + 2.166
In addition, the amount of protein in each well was measured using BCA Protein Assay Kit manufactured by PIERCE, and the amount of dopamelanin produced per unit protein was determined. The evaluation was performed by obtaining a relative value when the control value was 100. The results are shown in Table 1.

  As shown in Table 1, all the compounds of the present invention exhibited a high human epidermal melanocyte tyrosinase activity inhibitory effect.

  The tyrosinase activity inhibitor of the present invention effectively uses one or more compounds selected from nerol, vanillin, hydroxycitronellal, methyl heptine carboxylate, benzyl alcohol, methyl naphthyl ketone, rosephenone, and isobornyl acetate. Ingredients.

  Moreover, the above-mentioned tyrosinase activity inhibitor is mix | blended with a skin external preparation as a whitening active ingredient. The amount of the tyrosinase activity inhibitor to be blended in the external preparation for skin is not particularly limited as long as it is the amount that exhibits the whitening effect, but is preferably in the range of about 0.0001 to 0.5% by weight. When it mix | blends exceeding 0.5 weight%, there exists a possibility of giving an unpleasant fragrance to a skin external preparation. If it is less than 0.0001% by weight, it may be difficult to exert an effective whitening effect.

  Examples of the dosage form of the external preparation for skin of the present invention include cream, emulsion, foundation, pack, lotion, gel, solution, and stick. In addition, appropriate ingredients such as oils, moisturizers, thickeners, preservatives, emulsifiers, pigments, pH adjusters, other medicinal ingredients, ultraviolet absorbers, fragrances, etc. You can also

  In addition, in the skin external preparation in this invention, it is also possible to use together with another whitening active ingredient.

  Furthermore, the above-mentioned tyrosinase activity inhibitor is applied to foods as a food browning prevention active ingredient. When the tyrosinase activity inhibitor of the present invention is applied to foods and the like, an appropriate amount may be sprayed, applied, and immersed on the cut surface of fruits and vegetables such as vegetables and fruits.

  Details of the present invention will be described using examples.

[Examples 1 to 8, Comparative Example] Whitening serum
(1) Glyceryl tri-2-ethylhexanoate 7.8 (% by weight)
(2) Polyglyceryl distearate 2.4
(3) Hydrogenated soybean phospholipid 0.4
(4) Batyl alcohol 0.1
(5) Purified water Amount that makes the total amount 100
(6) Glycerin 7.5
(7) Xanthan gum 0.4
(8) Diethylenetriaminepentaacetic acid pentasodium 0.2
(9) 1,3-butylene glycol 2.5
(10) Methyl paraoxybenzoate 0.1
(11) Ethanol 4.0
(12) Fragrance 0.1
(13) Tyrosinase activity inhibitor shown in Table 2 Quantity production method shown in Table 2: The oil phase components (1) to (4) are mixed, dissolved and heated to 75 ° C. On the other hand, the aqueous phase components (5) to (10) are mixed and dissolved and heated to 75 ° C. Subsequently, after adding an oil phase component to the said water phase component and pre-emulsifying, it emulsifies uniformly with a homomixer. Cool with stirring and add (11) to (13) at 40 ° C. to mix and homogenize.

[Example 9] Cream for whitening
(1) 1,3-butylene glycol 10.00 (wt%)
(2) Methyl paraoxybenzoate 0.10
(3) Sucrose stearate 0.35
(4) Sodium N-stearoyl-L-glutamate 0.35
(5) Carboxyvinyl polymer (1 wt% aqueous solution) 2.00
(6) 2-Methacryloyloxyethyl phosphorylcholine
Butyl methacrylate copolymer liquid polyglyceryl monolaurate 0.50
(7) Purified water 65.96
(8) Squalane 3.00
(9) Octyldodecyl myristate 3.00
(10) Lipophilic glyceryl monostearate 3.00
(11) Beeslow 1.00
(12) Stearic acid 1.00
(13) Behenyl alcohol 2.50
(14) Hardened palm oil 2.00
(15) Jojoba oil 0.10
(16) Stearyl glycyrrhetinate 0.05
(17) Dimethylsiloxane / vinyldimethylsiloxane copolymer solution 1.00
(18) Ethanol 3.00
(19) Nellore 0.02
(20) Vanillin 0.02
(21) Fragrance 0.05
(22) Ascorbic acid phosphate magnesium salt 0.50
(23) Sodium citrate 0.50
Production method: The oily components (1) to (7) and the aqueous components (8) to (17) are mixed and homogenized, and heated to 75 ° C. After the oily component is added to the aqueous component and emulsified, the components (18) to (23) are added.

[Example 10] Whitening lotion
(1) Purified water 86.52
(2) Dipotassium glycyrrhizinate 0.05
(3) Polyethylene glycol (4000) 1.00
(4) Trimethylglycine 1.00
(5) Red algae extract 1.60
(6) Ascorbic acid phosphate sodium salt 2.00
(7) Sodium hydroxide (10 wt% aqueous solution) 2.20
(8) Sodium citrate 0.30
(9) Diethylenetriaminepentaacetic acid pentasodium (40% by weight aqueous solution) 0.20
(10) Hydroxycitronellal 0.01
(11) methyl heptine carboxylate 0.01
(12) Benzyl alcohol 0.01
(13) Methyl naphthyl ketone 0.01
(14) Rosephenone 0.01
(15) Ethanol 5.00
(16) Polyoxyethylene (60EO) hydrogenated castor oil 0.02
(17) Fragrance 0.01
(18) Methyl paraoxybenzoate 0.05
Production method: (1) to (18) are mixed and made uniform.

  About Example 1-Example 10 of this invention prepared by the said prescription, and the comparative example, the improvement effect of the pigmentation symptom was evaluated. The effect of improving pigmentation symptoms is a group of 20 female panelists with pigmentation symptoms such as remarkable stains and buckwheat, and each group is used twice a day for 2 months each day for each example or comparative example. Then, the state of skin pigmentation after 2 months was observed and evaluated in comparison with before use. The pigmentation state was evaluated according to the criteria shown in Table 3, and the average value of 20 people was calculated and shown in Table 4.

  As is clear from Table 4, in the examples using groups according to the present invention, marked improvement in pigmentation symptoms was observed in all groups, and only mild pigmentation was observed after the end of the use test. Symptoms were improved to a certain extent, and the whitening action was similar or higher than that of a comparative example in which sodium lactate, which has been widely known as a whitening agent, was blended.

  In Examples 1 to 10 of the present invention, no changes in the state of the preparation such as precipitation, separation, aggregation, odor change and discoloration of the components were observed during the use test period. Moreover, in each Example use group, the paneler which showed skin irritation reaction and skin sensitization reaction did not exist.

  Next, a 0.1% by weight ethanol solution of the tyrosinase activity inhibitor shown in Table 5 was prepared, sprayed on the surface of an apple that had been divided into eight parts, sprayed with a spray, and observed as it was at room temperature. did. As a result, browning was observed in Comparative Example 1 sprayed with ethanol, but no browning was observed in Examples 11 to 18 sprayed with the tyrosinase activity inhibitor of the present invention.

Claims (1)

A tyrosinase activity inhibitor comprising, as an active ingredient, one or more components selected from hydroxycitronellal, methyl heptine carboxylate, methyl naphthyl ketone, rosephenone, and isobornyl acetate .