JP3154834B2 - Tyrosinase inhibitor - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel tyrosinase inhibitor which is effective as a skin whitening agent and an anti-blackening agent for foods and the like.

[0002]

2. Description of the Related Art The blackening of skin due to spots, freckles and sunburn is due to the formation of melanin in melanocytes present in the epidermal basal cell layer of the skin.
Further, the blackening phenomenon of fresh foods such as seafood, fruits and vegetables is also due to the formation of melanin pigment. Regarding the biosynthesis of melanin, in the cytoplasmic granule melanosome in melanocytes, a pathway is known in which tyrosine is oxidized by tyrosinase, and melanin is biosynthesized via dopa, dopaquinone, and indolequinone. It has been known that melanin production is promoted by autoxidation caused by.

[0003] Therefore, prevention of spots, freckles and sunburn,
To prevent blackening of foods and the like, UV-A, B and C, which are harmful ultraviolet rays, are irradiated with a benzophenone derivative,
There are known a method of absorbing with an ultraviolet absorbing substance such as a benzotriazole derivative and a p-aminobenzoic acid derivative, and a method of controlling the production of a melanin pigment with a tyrosinase inhibitor such as kojic acid.

A 2-aryl-1,3-thiazolidine derivative having a chemical structure similar to that of the thiazolidine derivative according to the present invention has an inhibitory action on melanin production. However, the melanin production inhibitory action is not based on the tyrosinase inhibitory action but on the formation of a reaction product with dopaquinone. Further, the 2-aryl-1,3-thiazolidine derivative exhibits extremely low water solubility (Bi
ochim. Biophys. Acta, 1073 : 416-422, 1991).

On the other hand, the thiazolidine derivative according to the present invention has water solubility (J. Am. Chem. Soc. 70 : 3429-3431,
1948), it is not known to have a tyrosinase inhibitory effect.

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel tyrosinase inhibitor which is effective as a skin whitening agent and an anti-blackening agent for foods and the like.

[0007]

Means for Solving the Problems As a result of intensive studies, the present inventors have found that the thiazolidine derivative according to the present invention is:
It has been found to have a tyrosinase inhibitory action comparable to kojic acid.

That is, the present invention provides the following general formula (1)

[0009]

Embedded image

(Wherein R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group, R ₃ represents a hydrogen atom, a carboxyl group or an alkoxycarbonyl group, and X has 1 to 4 substituents. Which represents a pyridyl group which may be used as an active ingredient) and a salt thereof.

To produce the thiazolidine derivative according to the present invention, a known production method (J. Am. Chem. Soc. 70 : 3429-3) is used.
431, 1948).

The substituents in the thiazolidine derivative according to the present invention will be described below.

The alkyl group represented by R ₁ and R ₂ represents a linear or branched lower alkyl group. Examples of the linear or branched lower alkyl group include a methyl group, an ethyl group and a propyl group. Butyl group, pentyl group, hexyl group, heptyl group, isopropyl group and isobutyl group.

The alkoxycarbonyl group for R ₃ represents a lower alkoxycarbonyl group. Examples of the lower alkoxycarbonyl group include a methoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl group, a butoxycarbonyl group and a pentyloxycarbonyl group. Can be mentioned.

X represents a pyridyl group which may have 1 to 4 substituents. Examples of the substituent which may be substituted on the pyridyl group include methyl, ethyl, propyl, butyl, pentyl and the like. Groups, hexyl groups, heptyl groups, isopropyl groups, isobutyl groups, etc., alkyl groups, hydroxymethyl groups, hydroxyethyl groups, hydroxypropyl groups, etc., hydroxyalkyl groups, methoxy groups, ethoxy groups, propoxy groups, etc., and phosphono groups. Oxy groups and hydroxyl groups and the like can be mentioned, and as the pyridyl group,
Examples thereof include a 2-pyridyl group, a 3-pyridyl group, and a 4-pyridyl group, and a 4-pyridyl group is particularly preferable.

As a specific example of X, 2-methyl-3
-Hydroxy-5-hydroxymethyl-4-pyridyl group, 2-methyl-3-hydroxy-5-phosphonooxy-4-pyridyl group, 2-methyl-3-hydroxy-5
-Hydroxy-4-pyridyl group and the like.

Representative examples of the thiazolidine derivative according to the present invention include 2- (2-methyl-3-hydroxy-).
5-hydroxymethyl-4-pyridyl) -thiazolidine, 2- (2-methyl-3-hydroxy-5-hydroxymethyl-4-pyridyl) -4-carboxythiazolidine, 2- (2-methyl-3-hydroxy-5) -Hydroxymethyl-4-pyridyl) -5,5-dimethyl-4-carboxythiazolidine, 2- (2-methyl-3-hydroxy-5-phosphonooxy-4-pyridyl) -4-carboxythiazolidine, 2- ( 2-methyl-3-hydroxy-5-phosphonooxy-4-pyridyl) -thiazolidine, 2- (2-methyl-3-hydroxy-5-phosphonooxy-4-pyridyl) -4-ethoxycarbonylthiazolidine, 2 -(2-methyl-3-hydroxy-
5-hydroxy-4-pyridyl) -4-carboxythiazolidine, 2- (2-methyl-3-hydroxy-5-hydroxy-4-pyridyl) -thiazolidine and 2- (2
-Methyl-3-hydroxy-5-hydroxy-4-pyridyl) -4-ethoxycarbonylthiazolidine.

Examples of the salt of the thiazolidine derivative according to the present invention include pharmacologically acceptable salts such as hydrochloride, sodium salt, potassium salt and phosphate.

The inhibitory action of 2-aryl-3,4-thiazolidine derivative on melanin production, which is similar in chemical structure to the thiazolidine derivative according to the present invention, is presumed to be due to the thiol-type compound produced by hydrolysis of the thiazolidine derivative. (Biochim. Biophys. Acta, 1073 : 416
-422, 1991).

On the other hand, thiol compounds such as cysteine and glutathione are also known to inhibit tyrosinase. However, when thiol compounds use L-dopa as a substrate, lag time (reaction delay) occurs at the beginning of the reaction.
(Biochem. J., 235,
1991). However, since no lag time is observed in the thiazolidine derivative according to the present invention, the tyrosinase inhibitory mechanism is presumed to be different from thiol compounds such as cysteine and glutathione, and the thiazolidine derivative according to the present invention is derived from tyrosine. It is speculated that they inhibit the process of L-dopa.

The field of application of the tyrosinase inhibitor of the present invention is a skin whitening agent and a blackening inhibitor for foods and the like.

When the tyrosinase inhibitor of the present invention is used as a skin lightening agent, it is generally applied to the skin as an external preparation for the skin. A method is also possible.

Examples of the external preparation for skin include ointments, creams, emulsions, liniments and lotions.

These preparations for external use on the skin contain various components generally used for the thiazolidine derivative according to the present invention, for example, an aqueous component, a powder component, a humectant, a surfactant, a preservative, a thickener, an ultraviolet absorber, A perfume or the like can be blended and manufactured using a known formulation technique, but these various components can be used within a quantitative and qualitative range that does not impair the effects of the present invention.

When the tyrosinase inhibitor of the present invention is used as an external preparation for skin, the dosage and administration method of the external preparation are determined by adding the thiazolidine derivative according to the present invention to the preparation.
0.001 to 10% (W / W), preferably 0.1 to 10%
5% (W / W) is prepared to contain the face, neck, arms and hands, etc.
1 day a day on tan-prone or tanned areas
An appropriate amount may be applied once or several times.

When used as a blackening inhibitor for foods and the like, the thiazolidine derivative according to the present invention is used in an amount of 0.001 to 0.001.
-10% (W / V), preferably 0.01-5% (W / V)
/ V) An aqueous solvent is prepared so as to contain it, and when the food is fish and shellfish, an appropriate amount may be added to the feed and ingested, or the fish and shellfish may be injected, dipped, sprayed or the like in an appropriate amount. If the food is a fruit or vegetable, it may be sprayed or dipped in an appropriate amount.

Hereinafter, the present invention will be described in more detail with reference to Test Examples and Examples.

[0028]

[Test example]

1. Tyrosinase inhibitory action by ultraviolet absorbance method Mushroom-derived tyrosinase (Sigma)
Dissolve in 15M phosphate buffer (pH 6.8) and add about 128
The solution was adjusted to a concentration of 4 units / ml. The substrate solution was prepared by dissolving tyrosine and L-dopa in the phosphate buffer, and was prepared at 1.66.
The solution was adjusted to a concentration of mM. The sample solution is prepared by dissolving kojic acid and 2- (2-methyl-3-hydroxy-5-hydroxymethyl-4-pyridyl) -4-carboxythiazolidine (hereinafter abbreviated as compound X) in the phosphate buffer. , To a solution concentration of 0.01M.

First, 1800 μl of the phosphate buffer was mixed with 900 μl of a tyrosine or L-dopa substrate solution (this mixture was designated as solution A). At the same time, when tyrosine was used as a substrate, 50 μl of a phosphate buffer was added to 100 μl of a tyrosinase solution, and when L-dopa was used as a substrate, 50 μl of a tyrosinase solution was added to 50 μl of a tyrosinase solution.
Then, 200 μl of a 0.01 M sample solution (kojic acid solution or compound X solution) was mixed with the mixture (this mixture was designated as solution B). Solution A and solution B
After standing at 7 ° C. for 10 minutes, the two solutions were mixed, and a spectrophotometer (U
best-50, manufactured by JASCO Corporation, the absorbance (475 nm) was measured over time. For the control reaction solution without addition of the inhibitor, a phosphate buffer was used instead of the sample solution. The inhibition rate was calculated by the following formula using the slope obtained from the time-dependent change curve of each of the control reaction solution and the sample solution.

Inhibition rate (%) = (1−Slope of sample solution / Slope of control solution) × 100

[0031]

[Table 1]

2. Tyrosinase inhibitory action by oxygen electrode method 900 µl of a substrate solution of tyrosine or L-dopa was mixed with 1800 µl of the phosphate buffer (this mixed solution was referred to as solution A). At the same time, 100 μl of tyrosinase solution and 0.01 M specimen solution (kojic acid solution or compound X solution)
200 μl were mixed (this mixture was designated as solution B). After the solution A and the solution B were left at 37 ° C. for 10 minutes, both solutions were mixed and the amount of dissolved oxygen was measured (YSI 53
00 type biological oxygen monitor, manufactured by Yellow Springs). The inhibition rate was calculated from the oxygen consumption of each of the control reaction solution and the sample solution 5 minutes after the start of the reaction and the following formula.

Inhibition rate (%) = (1-oxygen consumption 5 minutes after sample solution / oxygen consumption 5 minutes after control reaction solution) × 10
0

[0034]

[Table 2]

[0035]

[Examples] Formulation examples are shown below. 1. Ointment An ointment was produced according to the following formulation by a conventional method.

White petrolatum 24.0 g Stearyl alcohol 24.0 g Propylene glycol 11.0 g Sodium lauryl sulfate 1.5 g 2- (2-Methyl-3-hydroxy-5-hydroxymethyl-4-pyridyl) -4-carboxythiazolidine 3 0.0 g methyl paraoxybenzoate 0.025 g propyl paraoxybenzoate 0.015 g purified water 100 g total

2. Lotion A lotion was prepared by a conventional method according to the following formulation.

Stearyl alcohol 2.0 g Light liquid paraffin 23.5 g Sodium lauryl sulfate 1.0 g Propylene glycol 11.0 g 2- (2-methyl-3-hydroxy-5-hydroxymethyl-4-pyridyl) -4-carboxythiazolidine 3.0 g Methyl parahydroxybenzoate 0.025 g Propyl paraoxybenzoate 0.015 g Purified water Total 100 g

3. Anti-fish blackening agent According to the following formula, an anti-fish blackening agent was produced.

L-ascorbic acid 0.6 g L-ascorbic acid 4.0 g lactose 13.0 g 2- (2-methyl-3-hydroxy-5-hydroxymethyl-4-pyridyl) -4-carboxythiazolidine 3.0 g Sorbitol 1.2g Alum 1.1g Purified water 100ml

[0041]

The thiazolidine derivative according to the present invention comprises:
It shows a tyrosinase inhibitory effect comparable to kojic acid. Also, kojic acid non-selectively inhibits the process involving two tyrosinases, L-dopa from tyrosine and dopaquinone from L-dopa, whereas the thiazolidine derivative according to the present invention inhibits the process from tyrosine to L-dopa. Selectively inhibits strongly.

Furthermore, the thiazolidine derivative according to the present invention is excellent in water solubility (Can. J. Chem. 69 : 1-7, 19).
91) It can be easily used as a skin lightening agent and an anti-blackening agent for foods and the like.

The thiazolidine derivatives of the present invention, since the reaction product of pyridoxal and amino acid vitamin B _6, because it is presumed that no toxic ones to the human body during the decomposition products, such as a human body and fish It is considered highly safe for animals.

Accordingly, the tyrosinase inhibitor of the present invention comprises
It is very excellent as a skin whitening agent and a blackening inhibitor for foods and the like.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61K 7/48 A61K 7/48 31/44 31/44 A61P 17/00 A61P 17/00 // C07D 417/04 C07D 417/04 (58) Field surveyed (Int. Cl. ⁷ , DB name) C12N 9/99 C07D 417/04 CA (STN) REGISTRY (STN)

Claims (4)

(57) [Claims] 1. General formula (1) (Wherein R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group, R ₃ represents a hydrogen atom, a carboxyl group or an alkoxycarbonyl group, and X may have 1 to 4 substituents. Tyrosinase inhibitor comprising a thiazolidine derivative represented by the following formula: 2. General formula (1) (Wherein R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group, R ₃ represents a hydrogen atom, a carboxyl group or an alkoxycarbonyl group, and X may have 1 to 4 substituents. A tyrosinase inhibitor comprising a thiazolidine derivative represented by the following formula: 3. A skin lightening agent comprising the tyrosinase inhibitor according to claim 1 or 2 as an active ingredient. 4. A blackening inhibitor comprising the tyrosinase inhibitor according to claim 1 or 2 as an active ingredient.