JPH0369322B2 - - Google Patents

Description

[Detailed description of the invention]

INDUSTRIAL APPLICATION FIELD The present invention relates to cosmetics for the purpose of, for example, controlling stains and freckles, preventing sunburn, and treating pigmentation disorders, etc.
This invention relates to tyrosinase activity inhibitors used in pharmaceuticals, etc. BACKGROUND ART Melanin is a black to brown ecological 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 the well-controlled and uniform formation of melanin is
It is preferable both for maintaining the normal functions of living organisms and from the viewpoint of cosmetics. However, local excessive melanin formation caused by external stimulation such as ultraviolet rays or metabolic abnormalities causes cosmetically unfavorable phenomena such as age spots, freckles, and hyperpigmentation. Although the true cause of such local hypermelaninosis is unknown, the phenomenon is attributed to hyperactivity of the enzyme tyrosinase involved in melanin formation. By inhibiting tyrosinase activity, tyrosinase activity inhibitors suppress such excessive melanin formation and promote whitening of the skin. Utilizing this skin whitening effect, tyrosinase activity inhibitors are used in cosmetics, pharmaceuticals, etc. for the purpose of controlling age spots and freckles, preventing sunburn, and treating hyperpigmentation disorders. Conventionally, crude drug extracts (Japanese Patent Application Laid-Open No. 79033/1983) have been used as tyrosinase activity inhibitors for the purpose of whitening effects.
No. 53-88333, No. 54-2344, No. 57-163307
), mixture of ascorbic acid and crude drug extract (JP-A-52-79032), mixture of ascorbic acid and aloin (JP-A-51-95140), ascorbic acid and 2,4,6-hydroxypropiophenone (Japanese Unexamined Patent Publications No. 51-101138), silk peptide compounds (Unexamined Japanese Patent Application Nos. 57-3827 and 57-40495),
Lipoic acid compounds (JP-A-57-123107), tiopronin-based compounds (JP-A-56-154409), glutathione-based compounds (JP-A-57-134410), pantethine-based compounds (JP-A-56-73012) ), pantetheine compounds (JP-A-57-7405, JP-A No. 59-36606)
No.), trobolone compounds (Japanese Patent Application Laid-open No. 56-26842,
56-147704, 56-147705), chromone compounds (JP-A-55-11410, JP-A-55-143908),
Flavonol compounds (JP-A No. 55-92305, same)
No. 55-111411, No. 55-157580, No. 58-131911
), pyrone compounds (JP-A-53-3538, JP-A No. 57)
-72978, 57-134409), kojic acid compounds (JP-A-53-6432, JP-A-56-7776, JP-A-56-77272)
No. 59-33207), and many others have been disclosed. Problems to be solved by the invention Conventionally known tyrosinase activity inhibitors are:
Some lack efficacy, some seem to have problems with stability and safety, and some are difficult to obtain. There is still no ideal solution that satisfies these requirements. Currently, tyrosinase activity inhibitors, while having excellent efficacy as the first priority, also have stability, safety, and
There is a demand for the emergence of an ideal product that also satisfies economic efficiency. Means for Solving the Problems In order to solve the above problems, the present inventors have extensively searched for substances that inhibit the melanin formation reaction from tyrosine by tyrosinase. , 4-methylenedioxyphenol and its ester derivatives and ether derivatives have been found to be highly effective in inhibiting tyrosinase activity, and have also demonstrated their stability, safety,
After thorough consideration of economic efficiency and the application of its whitening effect to cosmetics, pharmaceuticals, etc., we have confirmed that these compounds can constitute almost ideal tyrosinase activity inhibitors, and have developed the present invention. completed. That is, the present invention is based on the general formula () [In the formula, R is a hydrogen atom, ethyl, n-butyl, n
- from the group consisting of hexyl, cis-3-hexenyl, 2-methoxyethyl, benzyl, phenylethyl, cinnamyl, geranyl, prenyl, retinyl, tocopheryl, rosinyl, acetyl, butyryl, caproyl, oleoyl, benzoyl, cinnamoyl, abiethyl and retinoyl. Indicates at least one selected item. ] A tyrosinase activity inhibitor containing at least one 3,4-methylenedioxyphenol derivative represented by the following as an active ingredient. 3,4-Methylenedioxyphenol (referred to as Cezamol.Hereinafter referred to as Cezamol) has the structural formula of formula () This compound is naturally contained in small amounts in sesame oil along with sezamin, sezamorin, etc., and the purified product is a colorless crystal (melting point: 64°C) with a weak phenolic aroma. Although it is widely known that sezamol has an antioxidant effect, no literature has yet been found that reports that sezamol has an excellent tyrosinase activity inhibitory effect. Sezamol is naturally contained in sesame oil, but since it is in a trace amount, it is not a good idea to separate and purify it, and it is better to obtain sezamol by a synthetic method. Sezamol was synthesized by adding a peracetic acid solution containing a small amount of p-toluenesulfonic acid to piperonal and reacting at room temperature according to the method of Koden et al. [Rec.trav.chim., 55 , 815-20 (1936)]. After cooling, peracetic acid is decomposed with an alkali, unreacted piperonal is removed with a mixture of phenylhydrazine acetic acid, and then solvent extraction is carried out, thereby easily achieving a good yield. A specific example is as follows. Synthesis of Sezamol 420 g of 20% peracetic acid containing a small amount of p-toluenesulfonic acid is slowly added to 150 g of piperonal while stirring while maintaining the internal temperature at 30°C. After the reaction mixture is allowed to stand overnight to complete the reaction, most of the acetic acid is distilled off under reduced pressure. After adding alcoholic potassium hydroxide to decompose peracetic acid, 10 g of phenylhydrazine and 50 g of 4N acetic acid are added to remove unreacted piperonal. After making it acidic by adding dilute sulfuric acid, it is extracted with ether. The ether layer was washed with aqueous sodium bicarbonate and then with water, dried over Glauber's salt, and the ether was distilled off to obtain about 90 g of solid sezamol. This is re-purified from toluene. Sezamol has a phenolic hydroxyl group,
The skin irritation was low, and even when a 12.5% acetone solution of sezamol was applied to shaved guinea pigs, no erythema or redness was observed, and no sensitization was observed. 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, linear and cyclic terpene carboxylic acids, such as rosin acid,
Retinoic acid, etc. Also, the alcohols that participate in the formation of ether derivatives of sezamol include aliphatic saturated and unsaturated alcohols, such as ethanol, butanol, methyl cellosolve, hexenol, prenyl alcohol, aromatic alcohols, such as benzyl alcohol, phenethyl alcohol. etc., linear and cyclic terpene alcohols such as geraniol, retinol, modinol, abietinol, etc. Ester and ether derivatives of sezamol are:
It can be synthesized by conventional methods from sezamol and carboxylic acids or alcohols. For example, sezamol acetate is produced by adding acetic anhydride containing a small amount of phosphoric acid to sezamol, reacting at room temperature, adding water to separate the organic layer, and separating the organic layer.
Obtained by distillation. Sezamol prenyl ether can also be produced by adding prenyl chloride and a small amount of benzyltriethylammonium chloride to sezamol soda salt and stirring and refluxing the mixture for several hours.
It is obtained by separating the organic layer and distilling it.
Specific examples are as follows. Synthesis of sezamol acetate 30 g of acetic anhydride and 0.2 g of phosphoric acid were added to 20 g of sezamol, stirred at room temperature for 7 hours, and then left overnight.
Add 100ml of water, stir for 2 hours, take the separated organic layer, wash thoroughly with water, distill the organic layer, collect a fraction of Bb83°~84°/0.2mmHg,
g of sezamol acetate is obtained. Synthesis of Cezamol Prenyl Ether 126 g of 5% caustic soda solution was added to 21 g of Cezamol to make soda salt, and prenyl chloride 33
g and 5 g of benzyltriethylammonium chloride were added, and the mixture was stirred and refluxed at 70 to 75°C for 7 hours. After standing overnight, the separated organic layer was taken, washed with dilute hydrochloric acid and then water, and then distilled to Bb97° ~
Collect the 100°/0.3 mmHg fraction to obtain about 18 g of sezamol prenyl ether. When sezamol contains impurities, it may darken over time, but its ester and ether derivatives show almost no deterioration over time. Furthermore, when comparing the tyrosinase activity inhibitory effects of sezamol and its ester and ether derivatives, the derivatives tend to be slightly lower; however, when using them, it is important to consider not only the inhibitory effect but also the stability and safety of the substance. It is desirable to use the material by taking into consideration its properties, mixability with the base material, absorption into the skin, etc. Cezamol and its derivatives as tyrosinase activity inhibitors can be used in cosmetics and pharmaceuticals at an addition amount of 0.01 to 5% by weight to the base material, but in most cases, an addition amount of 0.01 to 0.2% by weight is used. A sufficient effect can be obtained. The addition method is by dissolving it in a hydrophilic solvent such as ethanol, glycerin, propylene glycol, or a lipophilic solvent such as triethyl citrate, benzyl benzoate, dioctyl phthalate, etc.
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. EXAMPLES The present invention will be explained in more detail with reference to Examples below, but the present invention is not limited to these Examples. Example 1 Tyrosinase activity inhibition effect of sezamol and its derivatives Take 2.0 ml of 0.1N phosphate buffer (PH7.0) containing 0.1% L-tyrosine and 0.002% copper sulfate, and add sezamol or its derivatives to this. each
Add 0.05 ml of ethanol solution containing 0.2%.
Next, 0.05 ml of an enzyme solution prepared by dissolving 10 mg of commercially available tyrosinase (manufactured by Sigma Chemical Co., 500 units per mg) in 10 ml of 0.1N phosphate buffer (PH.0) is added. The mixture was shaken at 37°C for 60 minutes, then held at 85°C for 5 minutes to stop the reaction, 2 ml of 0.1N phosphate buffer (PH7.0) was added, and the absorbance (OD) at a wavelength of 580 nm was measured. When the inhibition rate of the tyrosinase reaction was determined using the following formula, the results shown in Table 1 were obtained. Inhibition rate = (1 - OD when active inhibitor is added - blank OD / OD when active inhibitor is not added - blank OD) x 100

[Table] Example 2 Effect of inhibiting tyrosinase activity in melanoma cells B-16 melanoma cells, which are melanin-producing tumor cells, were inoculated into Eagle MEM medium (manufactured by Nippon Suisan) containing 20% fetal bovine serum, and then cezamol, A 0.5% dimethyl sulfoxide solution containing 0.2% each of sezamol retinoate and sezamol retinol ether was added, and 1/2 of the culture medium was refreshed every 2 days at 37°C and a carbon dioxide concentration of 5%. After 6 days of culture, cells were separated. Melanin content was determined by Uitztaker's method [Dev.
Biol., 8 , 99-127 (1963)], wavelength 400 nm.
It was expressed as the optical density (OD) per 10 ⁶ cells. Tyrosinase activity was determined by suspending the separated cells in a 0.001N caustic soda solution containing 0.5% sodium cholate and homogenizing the cells according to the method of Iwata et al. [Proc. Japan Acad., 56 , 562-567 (1980)]. After disrupting the cells, ultra-high-speed centrifugation (30,000 rpm, 20 minutes) was performed, and the resulting supernatant was ΔE/min per ¹⁰⁶ cells at a wavelength of 475 nm.
It was expressed as The results are shown in Table 2.

[Table] From Table 2, it can be seen that sezamol and its derivatives
It is known that it is absorbed into cells and acts effectively as a tyrosinase activity inhibitor within the cells. Example 3 Fading experiment on black goldfish Water was placed in tanks A, B, C, and D with a diameter of 30 cm and a depth of 20 cm.
10, and add 0.5% each of cezamol, cezamol acetate, and cezamol prenyl ether.
10 ml of ethanol solution was added, D was used as a control (no additives), and 3 black-eyed gilders were placed in each tank and reared. The water in the aquarium was refreshed every 3 days. All the goldfish in the aquariums to which the inhibitor had been added began to discolor from around the third week. Table 3 shows the degree of discoloration after 6 weeks and the number of chromatophores in the scales as determined by microscopic observation.

[Table] The results shown in Table 3 were sufficient to infer the skin whitening effect of sezamol and its derivatives. Example 4 cream agent (weight %) Monoglyceride 3.5 stearic acid 4.7 cethyl alcoholic 1.7 light flow flowing paraphrodes 14.0 isopropyl mirifin 3.0 stear lil alcohol 3.0 stearlil alcohol 3.2 butyl paraben 0.05 methyl paraben 0.05 triet Noamin 05 Trietnol Amin 0. 1 Pure Water 60.4 50 % Cesemall ethanol solution 1.0 Fragrance 0.3 100.0 First, mix the above formulation excluding glycerin, water, and fragrance, heat to about 70°C, stir thoroughly, keep at that temperature, add glycerin and water, and stir and mix further. do. Thereafter, the mixture was cooled with gentle stirring, and when the temperature dropped to about 50°C, a fragrance was added, and the mixture was further cooled to room temperature while stirring to obtain a cream. A cream sample prepared in the same manner as above was used as a blank, except that pure water was used instead of the 50% sezamol ethanol solution in the above formulation. In addition, from the above formulation, excluding the 50% Cezamol ethanol solution, 5% of Filtrasol-A (manufactured by Norda, USA), a commercially available sunscreen, was added instead, and 60.4% of pure water was added. A cream prototyped in the same manner as above was used as a positive control, except that the amount was set to 56.4% (hereinafter referred to as the control product). A sunburn test was conducted using these and a cream containing sezamol with the above formulation. Five Hartley guinea pigs weighing 350 to 400 g were used, and their shoulder blades were shaved and shaved. Divide the 3cm x 3cm hair removal area into 4 parts, and place 1
4 cm x 1 cm square test plots were prepared, two of which were filled with blanks, the remaining one was filled with cream containing sezamol (hereinafter referred to as the test product), and the remaining one was filled with a control product. It was applied at a concentration of 0.02 mg/cm ² . The unapplied area of the material was shielded from light. And with 5 lamps of National FL20SE, 10cm
The beam was irradiated for 10 minutes at a distance of 10 minutes (total energy amount was 1.8 Joules). As a criterion for evaluation, the following scores were given for erythema that appeared on the skin 24 hours and 48 hours after irradiation. Note that no evaluation was made regarding edema because only a few edema appeared overall. Rating: No erythema at all 0 Slight erythema 1 Obvious erythema 2 Moderate erythema 3 The results of the test based on this evaluation are shown in Table 4.

[Table] From the results in Table 4, it can be seen that Sezamol has a much superior effect (same effect with about 1/10th the amount) compared to commercially available sunscreens. Example 5 Ointment (wt%) Carite butter 40.0 Olive oil 20.0 Beeswax 20.0 Paraffin 19.9 Sezamol n-butyl ether 0.1 100.0 Each component was mixed according to the above recipe, heated to about 70°C and stirred well to make it sufficiently uniform. I poured it into a container and left it to cool to make an ointment. Example 6 Lotion (wt%) Ethanol 20.0 Propylene glycol 5.0 Glycerin 4.5 Methylparaben 0.1 Pure water 70.0 Sezamol acetate 0.1 Fragrance 0.3 100.0 A lotion was prepared by mixing each component according to the above recipe. Effects of the Invention The tyrosinase activity inhibitor of the present invention, which contains sezamol or its ester derivative or ether derivative as an active ingredient, substantially satisfies all of the requirements such as excellent efficacy, stability, safety, and economical efficiency, and has sufficient whitening effect. It has a sun protection effect and a pigmentation treatment effect.

Claims (1)

[Claims] 1 General formula () [In the formula, R is a hydrogen atom, ethyl, n-butyl, n
- selected from the group consisting of hexyl, cis-3-hexenyl, 2-methoxyethyl, benzyl, phenylethyl, cinnamyl, geranyl, prenyl, retinyl, tocopheryl, acetyl, butyryl, caproyl, oleoyl, benzoyl, cinnamoyl, and retinoyl. Indicates at least one type of thing. ] A tyrosinase activity inhibitor containing at least one 3,4-methylenedioxyphenol derivative represented by the following as an active ingredient. 2. The tyrosinase activity inhibitor according to claim 1, wherein the content of the 3,4-methylenedioxyphenol derivative is 0.01 to 5% by weight.