JPS6163609A - Inhibitor of tyrosinase activity - Google Patents

Abstract

PURPOSE:An inhibitor of tyrosinase activity useful as cosmetic, drug, etc. to inhibit formation of melanin, having excellent effect stability, etc., containing 3,4-methylenedioxyphenol, its ester derivative, or its ether derivative. CONSTITUTION:An inhibitor of tyrosinase activity containing 3,4-methylene dioxyphenol, its ester derivative, or ether derivative. The compound and its derivative can constitute an ideal inhibitor having excellent inhibitory effect on tyrosinase activity, satisfying further safety, stability, economic efficiency, etc. Addition of 0.01-0.2wt% compound shown by the formula or its derivative to cosmetic, or drug provides sufficient effects, the compound or its derivative is dissolved in a hydrophilic solvent such as ethanol, etc., or a lipophilic solvent such as triethyl citrate, etc., and used.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a tyrosinase activity inhibitor used in cosmetics, pharmaceuticals, etc. for the purpose of controlling age spots and freckles, preventing sunburn, and treating hyperpigmentation. .

BACKGROUND OF THE INVENTION Melanin is a black to brown biological pigment that is said to be produced by oxidative polymerization of tyrosine, and its formation is promoted by the enzyme tyrosinase. The formation of melanin itself is part of the body's defense mechanism, and well-controlled and uniform melanin formation is desirable both for maintaining the body's normal functions and from a cosmetic standpoint. Excessive melanin formation caused by external stimuli such as ultraviolet rays or metabolic abnormalities causes cosmetically unfavorable phenomena such as age spots, freckles, and hyperpigmentation.

The true cause of such local hypermelaninosis is unknown, but it is believed that it is caused by the completion of the activity of the enzyme tyrosinase, which is involved in melanin formation.Tyrosinase activity inhibitors reduce the activity of tyrosinase by 1 Tyrosinase activity inhibitors can control age spots and freckles, and prevent sunburn by utilizing their zero-force skin whitening effect, which suppresses such excessive melanin formation and increases the whiteness of the skin. It is used in cosmetics, pharmaceuticals, etc. for the treatment of pigmentation disorders, etc.

Conventionally, crude drug extract C has been used as a tyrosinase activity inhibitor for whitening effect.
-gg333, same! Gu 23 Gu Z issue.

j7-163307], a mixture of ascorbic acid and raw nicotine extract (JP-A Shoyo, No. 1-7903λ), a mixture of ascorbic acid and acoin (JP-A Sho! 1-q61)
Mixture of % ascorbic acid and Otsu-hydroxypropiophenone ('4 Kaisho 3l-IQ)
No. 13g),
No. 7, same! 2-g0g9! No.), Lipoic Acid Compound C Tokukaisho! 7-no, No. 23107), tiopronin compounds (
JP-A No. 67-1341'ljO), pantethine-based compounds (JP-A No. 67-1341'ljO), pantethine-based compounds (JP-A No. 67-1341'ljO), pantethine-based compounds (JP-A No. 67-1341'ljO), pantethine-based compounds J-7-7'lO! Same issue! 9-36
No. 404], Trobolone Compounds [JP-A-61. -21
．． g4t1, same jl>-/4t770'1.

66-/G No. 2705], Chromone Compounds (JP-A No. 63-//Z) No. 0, Same! ! -lg 39Dg)
, Flavonol Compound C JP-A-5! -9230J- No.

same! J'-///1li No.1, same! ! -167J'KO
JP-A-63-33'3g, JP-A No. 67-7277g, J'
7-/3'lg No. 09], kojic acid-based compounds (JP-A-Sho!
3-6g No. 32, same j6-7776, same! l, -77
No. 272, No. 69-33.207), and many others have been disclosed.

Problems to be Solved by the Invention Some of the conventionally known tyrosinase activity inhibitors lack significant efficacy, some seem to have problems with stability and safety, and some are difficult to obtain. There is still no ideal drug that satisfies all aspects of efficacy, stability, safety, economy, etc.

Currently, there is a demand for the emergence of ideal tyrosinase activity inhibitors that, while emphasizing excellent efficacy, also satisfy ζ et al.'s stability, safety, economic efficiency, etc.

Means for Solving the Problem The present inventors have attempted to solve the above problem by searching extensively for substances that inhibit the melanin formation reaction from tyrosine by tyrosinase.
We found out that different 3,4t-methylenedioxyphenol and its ester derivatives and ether derivatives are highly effective in inhibiting tyrosinase activity, and also thoroughly investigated their stability, safety, economic efficiency, etc. As a result of applying the whitening effect to cosmetics, pharmaceuticals, etc., it was confirmed that these compounds constitute almost ideal tyrosinase activity inhibitors, and the present invention was completed at the age of 0. The invention is 3. A tyrosinase activity inhibitor containing Z-methylenedioxyphenol, or its ester derivative or ether derivative as an active ingredient.

3. Z-methylenedioxyphenol (referred to as sezamol) is a compound represented by the Mizozo formula of the formula (E), and naturally exists in sesame oil as sezamin,
It is contained in trace amounts along with sezamolin, etc., and the purified product is a colorless crystal with a weak phenolic aroma (melting point, 4 gtl?
). It is widely known that cezamol has antioxidant effects, but cezamol has an excellent tyrosinase activity inhibitory effect 'Y! I have yet to find any literature that reports that this is the case.

Sezamol is naturally contained in sesame oil, but it is only in trace amounts, so it is not a good idea to separate and purify it. CRec, trav, c
him, +ta J-, gj! 20 (/93A)), peracetic acid containing a small amount of p-toluenesulfonic acid was added to biveronal.

After reacting at room temperature, peracetic acid is decomposed with an alkali, unreacted piveronal is removed with a mixture of phenylhydrazine and acetic acid 17'1, and solvent extraction is performed to easily improve the yield ( Let's do it.To give you a concrete example,
It is as follows.

Sezamol's synthetic piveronal IjQ], a small amount "p -) /l/
Add 20% peracetic acid containing efsulfonic acid slowly while stirring while keeping the internal temperature at 30°C. After leaving the reaction mixture overnight to complete the reaction, remove most of the acetic acid under reduced pressure. After adding alcoholic potassium hydroxide to this to decompose peracetic acid, 10 ml of phenylhydrazine, uM, and constant vinegar es were added.

to remove unreacted biperonal. After making it acidic by adding dilute sulfuric acid, it is extracted with ether. The ether layer was washed with sodium bicarbonate solution and then with water.

After drying with Glauber's salt, the ether is distilled off to obtain about 90 iP of solid sezamol, which is purified by recrystallization from toluene.

Although Cezamol has a phenolic hydroxyl group, it has low irritation to the skin (even when a 72.s% acetone solution of Cezamol was applied to a guinea pig with prickly hair, no erythema or redness was observed, and no sensitization was observed. I couldn't.

The carboxylic acids that participate in the formation of the ester derivatives of sezamol used as active ingredients in the present invention include aliphatic saturated and unsaturated carboxylic acids such as acetic acid, butyric acid, caproic acid, oleic acid, etc., aromatic carboxylic acids such as benzoic acid, etc. , cinnamic acid, and chain and cyclic terpene carboxylic acids such as rosin acid and retinoic acid.

Alcohols that also participate in the formation of ether derivatives of sezamol include aliphatic saturated and unsaturated alcohols, such as ethanol, butanol, methyl selonulf, hexenol, prenyl alcohol, etc., aromatic alcohols, such as evabenzyl alcohol, phenethyl alcohol, etc. , linear and cyclic terpene alcohols, such as hakelaniol and retinol.

Ester and ether derivatives of sezamol can be synthesized from sezamol and carboxylic acids or alcohols by conventional methods. For example, sezamol acetate can be obtained by adding acetic anhydride containing a small amount of phosphoric acid to sezamol, reacting at room temperature, adding water to separate the organic layer, fractionating the organic layer, and distilling it. Sezamol prenyl ether can be obtained by adding prenyl chloride and a small amount of benzyltriethylammonium chloride to sezamol soda salt and stirring and refluxing the mixture for several hours. After one reaction, one layer is separated and distilled.

A specific example is as follows.

Synthesis of sezamol acetate Sezamol, 209-, acetic anhydride 30 iP, phosphoric acid 0.2
After stirring at room temperature for 7 hours, the device was stirred for 7 hours.
Add 8'l of water, stir for 2 hours, take the separated organic layer, wash thoroughly with water, and distill one layer to yield Bb g j°~A'4t'/0.2mmH!;P
Collect the fractions to obtain about 23% of sezamol acetate 0 Synthesis of sesamol brenyl ether Sezamol, l/9-! % caustic soda [/koro? is added to make soda salt, and to this prenyl chloride 33),
Benzyltriethylammonium chlorite"t,
Add y and stir and reflux at 20-2 IC for 7 hours. −
After standing overnight, remove the separated organic layer.

After washing with dilute hydrochloric acid and then water, it was distilled to 97°~/DO°10.3mmH? Collect about 1 kg of fractions.
Cezamol Brenyl A-f) L”%: (G Le.

Almost no deterioration over time is observed in chilled conductors.

Furthermore, when comparing the tyrosinase activity inhibitory effects of sezamol and its ester and ether derivatives, the derivatives tend to be slightly lower. It is preferable to use cezamol and its derivatives as tyrosinase activity inhibitors in cosmetics and pharmaceuticals by taking into consideration the miscibility with the skin, absorption into the skin, etc. Although it can be used at -3% by weight, in many cases sufficient effects can be obtained by adding 0.01-0.2% by weight.

The addition method is hydrophilic @ agent such as ethanol, glycerin, propylene glycol, triethyl ntrate, etc.
This is done by dissolving it in a lipophilic solvent such as benzyl benzoate or dioctyl phthalate.

When the base material already contains the above-mentioned solvents, oils and fats, surfactants, and other components that easily dissolve sezamol and its derivatives, they can be added directly.

The present invention will be explained in more detail by giving examples below, but the present invention is limited only to these examples.
Et, no.

Example row 1 Tyrosinase activity inhibition effect of sezamol and its derivatives 0, no% L-tyrosine and o - o o, 2% F>H
e Q, l normal phosphate buffer containing copper (pH 7.0
) and 0, and added sezamol or a derivative thereof to 0. Ethanol solution containing 0%, osmt
Add. Then, commercially available tyrosinase (Sigma Chemical Co., Ltd. !00 units per 1 mg of MI)/(7m9 Q, 1 normal phosphate buffer fi (pH 7, o)/) was added.

0.0% of the commonly understood enzyme solution. Add Ojmi. A mixture? After shaking at 2C for 60 minutes, transfer to gIC! Hold for 1 minute to stop the reaction, add Q and l normal phosphate buffer (pH 7,
0) Add 2rni and absorbance K at wavelength 5g o nrn
(OD ) was measured and the inhibition rate of the tyrosinase reaction was determined using the following formula, and the results shown in Table 1 were obtained.

Inhibition rate = Table 1 Example 2 Effect of inhibiting tyrosinase activity in melanoma cells B-/6 melanoma cells, which are melanin-producing tumor cells, were inoculated into Eagle MEM medium (manufactured by Nippon Suisan) containing 20% fetal bovine serum. To this, a dimethyl sulfoxide bath containing 0.2% of each of Sezamol, Sezamol retinoate, and Sezamol retinol ether/fjlo, s% was added and treated under conditions of %32C1 carbon dioxide concentration!% every - day. After culturing for 6 days while renewing the medium, the cells were separated and the melanin content was determined by Reitzter's method [Dev.

Biol,, g, 99~/27 (/wa63)]
It was expressed as the absorbance (OD) per 7011 cells at a wavelength of 0 nm. In addition, tyrosinase activity was determined by Iwata et al. 0 method CProc, Japan Acad.
s b, s A x ~J[7(/9KO)], call the separated cells to 0. ! After suspending the cells in 0,001N caustic soda solution containing
000rpm.

20 minutes), and the resulting supernatant liquid has a wavelength of 2! nm
Expressed as Δ/min of cells/min.

The results are shown in Table 2.0 From Table 2, it is also called sezamol and its derivatives.

It is known that it is absorbed into cells and acts effectively as a tyrosinase activity inhibitor within the cells.
Add 1 jt of water) to this, add each O, S% ethanol solution of sezamol, sezamol acetate, sezamol prenyl ether [/01+1J], use D as control (no additives), and add 3 black eyelets to each aquarium. They were reared in t each. The water in the aquarium was refreshed every 3 days. Table 3 shows the degree of discoloration of the mouth of the goldfish in the aquarium containing the inhibitor, which began to discolor from around the third week, and the number of chromatophores in the scales observed under a microscope.

Table 3 12: It was enough to make people guess 0 Example 4 Cream (% by weight) Stearic acid monoglyceride 3. j Stearic acid, 7 cetyl alcohol, +1/', light liquid paraffin ivy, O isopropyl myristate 3.
0 Glycerin g・0 Stearyl alcohol 3.2 Butylbarape 10 01 Methylparaben D Otatriethanolamine 0.1 Pure water
604t j Easy Cezamol Ethanol Liquid 1.0 Fragrance 0.3 First, mix the above formulation excluding glycerin, water, and fragrance, stir thoroughly while heating to about 747'C, and keep at that temperature. Add glycerin and water, and mix again. Then, cool while continuing to gently stir.
When the temperature reached approximately OC, fragrance was added, and the mixture was cooled to room temperature while stirring to obtain a cream.

Example 5 Ointment (wt%) Karite butter Ri00 olive oil
200 beeswax
20 0 paraffin 1
9 Tasezamol n-butyl ether o
, 1. Each component was mixed according to the above recipe, heated to about 20C, stirred thoroughly, and made sufficiently uniform.The mixture was then poured into a container and left to cool to prepare an ointment.

One piece X example 6 makeup water (Weight section) Ethanol 2o.

Propylene glycol j, 0 glycerin,! Methylparaben
D, / pure water
200 Sezamol Acetate O,l Fragrance 0
．． 37Qσ0 Mix each component according to the above recipe and apply lotion (Tsuda〇Effects of the Invention The present invention's sezamol or its ester derivative properties are completely satisfied, and the skin whitening effect and sunburn prevention effect are completely satisfied. 1 has a therapeutic effect on pigmentation syndrome.

Handbook continuation supplementary book July 3, 1985

Claims (1)

[Claims] A tyrosinase activity inhibitor containing 3,4-methylenedioxyphenol, or its ester derivative or ether derivative as an active ingredient.