JP2011140447A - Dopa oxidase activity inhibitor and whitening agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a DOPA oxidase activity inhibitor, and to provide a whitening agent. <P>SOLUTION: The DOPA oxidase activity inhibitor contains a compound represented by general formula (1) or a salt thereof as an active ingredient. The whitening agent contains the compound or the salt thereof as an active ingredient. In the formula, X is hydroxy or carboxy; R is alkyl, alkenyl, aryl or aralkyl, which may be substituted; with the proviso that when X is carboxy, the total number of carbons in the group of R is ≥4; and n is 1 or 2. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a dopa oxidase activity inhibitor and a whitening agent.

  Pigmentation after sunburn, spots and freckles generally result from increased melanin production by activation of pigment cells (melanocytes) in the skin due to irritation caused by ultraviolet exposure to the skin, hormonal abnormalities or genetic factors It is believed that. In vivo, pigment melanin is biosynthesized from the precursor tyrosine by the action of the enzyme tyrosinase in melanosomes in pigment cells (melanocytes). It has been reported that when a mutation occurs in tyrosinase, an enzyme involved in melanin biosynthesis, the formation of melanin pigments in the skin and hair becomes abnormal (see, for example, Non-Patent Document 1).

  Tyrosinase, which plays an important role in melanin biosynthesis, is thought to be useful as a target substance for whitening materials, and the search for whitening materials focusing on tyrosinase has been made. Tyrosinase specifically has tyrosine hydroxylase activity, dopa oxidase activity, and DHI activity, and catalyzes a melanin synthesis reaction using tyrosine and dopa as precursors. Therefore, when examining tyrosinase enzyme activity, for example, dopa oxidase activity can be used as an index. Actually, dopa oxidase activity is also used when evaluating a melanin production inhibitory material having an inhibitory action on tyrosinase enzyme activity. It is used as an index (see, for example, Non-Patent Document 2). According to this method, a substance that suppresses the production of melanin, which is the final biosynthetic product, can be evaluated by suppressing dopa oxidase activity in melanocytes.

  Thus, conventionally, the search of the substance which can inhibit the activity of tyrosinase and suppress melanin production, the substance which can reduce the produced melanin, etc. was made | formed, and use of these substances as a whitening component has been examined. For example, it has been reported that ascorbic acid, arbutin, kojic acid, glutathione and the like have the action (for example, see Non-Patent Document 3).

King RA. Oetting WS. Hearing VJ. In Metabolic bases of inherited disease (Scriver CR. Beaudet AL. Sly WS. Valle D., eds.), McGraw-Hill, New York, 4353-4392, 1995 Wrathall JR. et al., JCB 1973 57: 406-423 Whitening strategy (Nanedo) IV. , Whitening agent pharmacology and clinic, p95-115

  This invention makes it a subject to provide the dopa oxidase activity inhibitor which can suppress dopa oxidase activity effectively. Moreover, this invention makes it a subject to provide the whitening agent which suppresses melanin production by suppressing dopa oxidase activity.

  In view of the above-mentioned problems, the present inventors have intensively studied to search for a new material having a whitening effect. As a result, it was found that the compound represented by the following general formula (1) has a dopa oxidase activity inhibitory action, and the knowledge that the compound is useful as a novel whitening component was obtained. The present invention has been completed based on this.

  That is, this invention provides the dopa oxidase activity inhibitor which contains the compound or its salt represented by following General formula (1) as an active ingredient.

（式中、Ｘは水酸基又はカルボキシル基を表す。Ｒはアルキル基、アルケニル基、アリール基又はアラルキル基を表し、これらの基は置換されていてもよい。ただし、Ｘがカルボキシル基の場合、Ｒの基は総炭素数４以上である。ｎは１又は２を表す。）

(In the formula, X represents a hydroxyl group or a carboxyl group. R represents an alkyl group, an alkenyl group, an aryl group or an aralkyl group, and these groups may be substituted. However, when X is a carboxyl group, R represents Group having 4 or more carbon atoms in total. N represents 1 or 2.)

  Moreover, this invention provides the whitening agent which contains the compound or its salt represented by the said General formula (1) as an active ingredient.

  The dopa oxidase activity inhibitor and whitening agent of the present invention contain, as an active ingredient, the compound represented by the general formula (1) having an excellent dopa oxidase activity inhibitory action or a salt thereof. Therefore, the dopa oxidase activity inhibitor and whitening agent of the present invention can suppress dopa oxidase activity, suppress melanin produced by tyrosinase, and prevent or prevent stains, buckwheat, sunburn pigmentation, and the like. It can be.

Hereinafter, the present invention will be described in detail.
The dopa oxidase activity inhibitor and the whitening agent of the present invention contain a compound represented by the following general formula (1) or a salt thereof as an active ingredient.

  In general formula (1), X represents a hydroxyl group or a carboxyl group. R represents an alkyl group, an alkenyl group, an aryl group or an aralkyl group, and these groups may be substituted. However, when X is a carboxyl group, the R group has 4 or more carbon atoms in total. n represents 1 or 2.

The alkyl group represented by R may be linear, branched or cyclic. In addition, the cyclic alkyl group includes a monocyclic, polycyclic or condensed cyclic group.
When X is a hydroxyl group, the alkyl group represented by R preferably has 1 to 30 carbon atoms, more preferably 1 to 25 carbon atoms, and particularly preferably 1 to 20 carbon atoms. . Specifically, methyl, ethyl, propyl, isopropyl, linear or branched butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl groups as linear or branched alkyl groups , An undecyl group, a dodecyl group, a tridecyl group, a tetradecyl group, a pentadecyl group, a hexadecyl group, a heptadecyl group, an octadecyl group, and the like, and examples of the cyclic alkyl group include cycloalkyl groups such as a cyclopentyl group, a cyclohexyl group, and a cycloheptyl group.
When X is a carboxyl group, the alkyl group represented by R has 4 or more carbon atoms, preferably 4 to 30 carbon atoms, more preferably 4 to 25 carbon atoms, and a carbon number of 4 to 20 is particularly preferable. Specifically, linear or branched butyl group, pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, undecyl group, dodecyl group, tridecyl group, tetradecyl group, pentadecyl group, hexadecyl group, heptadecyl group Group, octadecyl group and the like, and examples of the cyclic alkyl group include cycloalkyl groups such as cyclopentyl group, cyclohexyl group and cycloheptyl group.

The alkenyl group represented by R may be linear, branched or cyclic. In addition, the cyclic alkenyl group includes a monocyclic, polycyclic or condensed cyclic group.
When X is a hydroxyl group, the alkenyl group represented by R preferably has 1 to 30 carbon atoms, more preferably 1 to 25 carbon atoms, and particularly preferably 1 to 20 carbon atoms. . Specific examples of such an alkenyl group include an alkenyl group having one or more unsaturated groups such as a double bond in the specific examples of the alkyl group shown above where X is a hydroxyl group.
When X is a carboxyl group, the alkenyl group represented by R has 4 or more carbon atoms, preferably 4 to 30 carbon atoms, more preferably 4 to 25 carbon atoms, and a carbon number of 4 to 20 is particularly preferable. Specific examples of such an alkenyl group include alkenyl groups having one or more unsaturated groups such as double bonds in the specific examples of the alkyl group shown as the case where X is a carboxyl group.

The aryl group represented by R includes monocyclic, polycyclic or condensed cyclic groups, preferably 6 to 30 carbon atoms, more preferably 6 to 25 carbon atoms, Is particularly preferably 6-20. Specific examples include a phenyl group and a naphthyl group.
Examples of the aralkyl group represented by R include those in which the aryl group is monocyclic, polycyclic, or condensed cyclic, and preferably has 7 to 30 carbon atoms and 7 to 25 carbon atoms. More preferably, the number of carbon atoms is particularly preferably 7-20. Specific examples include a benzyl group and a phenethyl group.

  The group represented by R described above may be substituted. Examples of the substituent include a carboxyl group, an alkyl group (for example, an alkyl group exemplified as the above R), a hydroxyl group, an alkoxyalkyl group, and the like.

  In general formula (1), n represents 1 or 2. When X is a hydroxyl group, n is preferably 2. In addition, when X is a carboxyl group, n is preferably 1.

  In the present invention, the compound represented by the general formula (1) is preferably a compound represented by the following general formula (2) or a compound represented by the following general formula (3).

In General Formula (2), R ¹ represents an alkyl group, an alkenyl group, an aryl group, or an aralkyl group, and these groups may be substituted.
The alkyl group, alkenyl group, aryl group or aralkyl group represented by R ¹ is synonymous and synonymous with the alkyl group, alkenyl group, aryl group or aralkyl group represented by R in the general formula (1). Is the same.

In General Formula (3), R ² represents an alkyl group having 4 or more carbon atoms, an alkenyl group having 4 or more carbon atoms, an aryl group, or an aralkyl group, and these groups may be substituted.
The alkyl group, alkenyl group, aryl group or aralkyl group represented by R ² is synonymous and synonymous with the alkyl group, alkenyl group, aryl group or aralkyl group represented by R in the general formula (1). Is the same.

  Specific examples of the compounds represented by the general formulas (1) to (3) are shown below, but the compounds used in the present invention are not limited to these exemplified compounds.

  Moreover, the dopa oxidase activity inhibitor and whitening agent of this invention may contain the salt of the compound represented by the said General formula (1) as an active ingredient. That is, in the compound represented by the general formula (1), an anion derived from a carboxyl group or a hydroxyl group represented by X may form a salt with a cation as an active ingredient. Such a salt is not particularly limited, and examples thereof include alkali metal or alkaline earth metal salts such as sodium, potassium and calcium, amphoteric metal salts such as aluminum and zinc, amino acid salts and amine salts. Specifically, the salts of the exemplified compounds 1 to 17 shown above can be used.

  It does not specifically limit as a manufacturing method of the compound represented by the said General formula (1), or its salt, It can synthesize | combine by a normal chemical synthesis method etc. For example, it can be synthesized by a reaction between p-aminomethylbenzoic acid and a carboxylic acid chloride, or a reaction between tyramine and a carboxylic acid chloride. In addition, p-aminomethylbenzoic acid having a protecting group or tyramine having a protecting group is condensed with a corresponding carboxylic acid or a derivative thereof (such as a carboxylic acid ester or a carboxylic acid chloride), and then the protecting group is removed from the product. Can also be synthesized. As a specific synthesis method, a method shown in Examples described later can be used, but the present invention is not limited thereto.

As demonstrated in the below-mentioned Example, the compound or its salt represented by the said General formula (1) suppresses dopa oxidase activity effectively. Therefore, the whitening agent of the present invention containing the compound or a salt thereof can suppress tyrosinase activity, suppress melanin production in the skin, and exhibit a whitening effect. The compound represented by the general formula (1) or a salt thereof can also be used as a dopa oxidase activity inhibitor or a tyrosinase activity inhibitor.
In the present invention, “whitening (action)” refers to suppressing the formation of melanin, returning to the original transparent skin color without excess melanin, or preventing pigmentation such as blackening of the skin or spots and freckles. Mean to suppress.

In the dopa oxidase activity inhibitor or whitening agent of the present invention, the compound represented by the general formula (1) or a salt thereof may be used alone, or two or more kinds thereof may be mixed and used.
In the present invention, the compound represented by the general formula (1) or a salt thereof may be used as it is as a dopa oxidase activity inhibitor or a whitening agent. In addition, an appropriate liquid or solid excipient or extender such as titanium oxide, calcium carbonate, distilled water, lactose, starch and the like may be added to the compound or a salt thereof. In this case, the amount of the compound represented by the general formula (1) or a salt thereof is not particularly limited, but 0.0001 to 20% by mass of the compound represented by the general formula (1) or a salt thereof is included. Is preferably included, more preferably 0.001 to 10% by mass, and particularly preferably 0.005 to 5% by mass.

When the compound represented by the general formula (1) or a salt thereof is used as a whitening agent, other medicinal components may be used in addition to the compound. For example, other whitening agents, moisturizers, antioxidants, ultraviolet absorbers, surfactants, thickeners, colorant types, and the like can be added.
When the whitening agent of the present invention is used as a composition together with other medicinal ingredients, the amount of the compound represented by the general formula (1) or a salt thereof is not particularly limited, but is represented by the general formula (1). The compound or a salt thereof is preferably contained in an amount of 0.0001 to 20% by mass, more preferably 0.001 to 10% by mass, and particularly preferably 0.005 to 5% by mass.

The dopa oxidase activity inhibitor and whitening agent of the present invention can be used in the form of a skin external preparation. “Skin external preparation” means those applied to the skin as skin cosmetics, external pharmaceuticals, quasi-drugs, etc., and their dosage forms are aqueous solutions, solubilization systems, emulsification systems, powder systems, gels It can take a wide variety of forms, such as systems, ointments, creams, water-oil two-layer systems, water-oil-powder three-layer systems. Examples thereof include face wash, lotion, milky lotion, cream, gel, essence (beauty serum), pack, mask, foundation, ointment, sheet-like product, and the like.
When used in the form of a skin external preparation, in addition to the compound represented by the general formula (1) or a salt thereof, components used for normal skin external preparations such as surfactants, oily substances, and polymer compounds , Preservatives, medicinal ingredients other than those described above, powders, ultraviolet absorbers, dyes, fragrances, emulsion stabilizers, pH adjusters, and the like can be appropriately mixed. The amount of the external preparation for skin containing the compound represented by the general formula (1) or a salt thereof varies depending on the content of the active ingredient. For example, in the case of cream or ointment, per 1 cm ^{2 of} the skin surface In the case of 0.001 to 1000 μg and a liquid preparation, it is also preferable to use 0.001 to 1000 μg.

  EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited to this.

Synthesis Example Exemplified Compounds 1 to 17 and Comparative Compounds A and B described below were respectively synthesized by the following synthesis method.

1. Synthesis of Comparative Compound A 4- (Nt-butoxycarbonyl) aminomethylbenzoic acid methyl ester (0.5 g, 1.88 mmol) was dissolved in 4M hydrochloric acid / dioxane solution (15 mL) and stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure. The residue was dissolved in chloroform (15 mL), and while stirring at 0 ° C., acetic anhydride (3.76 mmol) and triethylamine (5.64 mmol) were added thereto, and then stirring was continued overnight at room temperature. After completion of the reaction, 2N-hydrochloric acid was added to acidify the system, the solvent was distilled off under reduced pressure, the residue was purified by silica gel column chromatography, and 4-acetylaminomethylbenzoic acid methyl ester (0.36 g). Yield 92.3%) as an intermediate. Next, the intermediate (0.32 g, 1.54 mmol) was dissolved in methanol (20 mL), and an aqueous sodium hydroxide solution (sodium hydroxide 3.08 mmol, water 10 mL) was added thereto, and the mixture was stirred at 60 ° C. for 1.5 hours. Stir. After cooling to room temperature, 2N-hydrochloric acid was added to acidify the system, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography to obtain Comparative Compound A (0.29 g, intermediate). Yield 96.7%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 1.88 (s, 3H), 4.30 (d, 2H, J = 6.02 Hz), 7.31 (d, 2H, J = 8.00 Hz), 7.88 (d, 2H , J = 8.00Hz), 8.43 (t, 1H, J = 5.93Hz), 13.05 (br, 1H)

2. Synthesis of Comparative Compound B Comparative Compound B was synthesized according to the synthesis method of Comparative Compound A using n-butanoic acid chloride instead of acetic anhydride (yield 42.0%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 0.86 (t, 3H, J = 7.38 Hz), 1.54 (m, 2H), 2.12 (t, 2H, J = 7.34 Hz), 4.31 (d, 2H , J = 5.83Hz), 7.32 (d, 2H, J = 8.00Hz), 7.88 (d, 2H, J = 8.09Hz), 8.38 (t, 1H, J = 5.93Hz), 13.09 (br, 1H)

3. Synthesis of Exemplified Compound 1 Exemplified Compound 1 was synthesized according to the synthesis method of Comparative Compound A using isobutanoic acid chloride instead of acetic anhydride (yield 83.9%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 0.88 (t, 6H, J = 6.40 Hz), 1.96-2.05 (m, 3H), 4.32 (d, 2H, J = 6.02 Hz), 7.35 (d , 2H, J = 8.19Hz), 7.88 (d, 2H, J = 8.28Hz), 8.40 (t, 1H, J = 5.93Hz), 12.91 (br, 1H)

4). Synthesis of Exemplified Compound 2 Exemplified Compound 2 was synthesized according to the synthesis method of Comparative Compound A using cyclohexanecarboxylic acid chloride instead of acetic anhydride (yield 56.7%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ1.11-1.27 (m, 3H), 1.31-1.40 (m, 2H), 1.59-1.64 (m, 1H), 1.68-1.75 (m, 4H), 2.14-2.20 (m, 1H), 4.30 (d, 2H, J = 6.02Hz), 7.32 (d, 2H, J = 8.38Hz), 7.88 (d, 2H, J = 8.28Hz), 8.31 (t, 1H , J = 6.02Hz), 12.90 (br, 1H)

5. Synthesis of Exemplified Compound 3 Exemplified Compound 3 was synthesized according to the synthesis method of Comparative Compound A using benzoyl chloride instead of acetic anhydride (yield 83.4%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ4.53 (d, 2H, J = 5.83 Hz), 7.38 (d, 2H, J = 8.00 Hz), 7.48 (m, 2H), 7.55 (m, 1H ), 7.87-7.93 (m, 4H), 9.13 (t, 1H, J = 6.02Hz), 13.12 (br, 1H)

6). Synthesis of Exemplified Compound 4 Exemplified Compound 4 was synthesized according to the synthesis method of Comparative Compound A using 2-ethylhexanoic acid chloride instead of acetic anhydride (yield 92.7%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 0.79 (t, 3H, J = 7.39 Hz), 0.83 (t, 3H, J = 7.34 Hz), 1.10-1.40 (m, 6H), 1.42-1.51 (m, 2H), 2.10 (m, 1H), 4.32 (d, 2H, J = 5.83Hz), 7.32 (d, 2H, J = 8.00Hz), 7.88 (d, 2H, J = 8.19Hz), 8.43 (t, 1H, J = 6.02Hz), 13.12 (br, 1H)

7). Synthesis of Exemplified Compound 5 Exemplified Compound 5 was synthesized according to the synthesis method of Comparative Compound A using 3-phenylpropanoic acid chloride instead of acetic anhydride (yield 88.7%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 2.47 (t, 2H, J = 7.62 Hz), 2.85 (t, 2H, J = 7.62 Hz), 4.30 (d, 2H, J = 5.83 Hz), 7.18-7.23 (m, 5H), 7.25-7.30 (m, 2H), 7.83 (d, 2H, J = 8.09Hz), 8.41 (t, 1H, J = 6.02Hz), 13.01 (br, 1H)

8). Synthesis of Exemplified Compound 6 Exemplified Compound 6 was synthesized according to the synthesis method of Comparative Compound A using n-octanoic acid chloride instead of acetic anhydride (yield 69.8%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 0.85 (t, 3H, J = 6.97 Hz), 1.17-1.29 (m, 8H), 1.51 (m, 2H), 2.14 (t, 2H, J = 7.34Hz), 4.31 (d, 2H, J = 5.93Hz), 7.34 (d, 2H, J = 8.28Hz), 7.88 (d, 2H, J = 8.28Hz), 8.39 (t, 1H, J = 5.93Hz) ), 12.91 (br, 1H)

9. Synthesis of Exemplified Compound 7 Exemplified Compound 7 was synthesized according to the synthesis method of Comparative Compound A using n-nonanoic acid chloride instead of acetic anhydride (yield 90.2%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 0.85 (t, 3H, J = 7.06 Hz), 1.17-1.30 (m, 10H), 1.51 (m, 2H), 2.14 (t, 2H, J = 7.34Hz), 4.31 (d, 2H, J = 5.83Hz), 7.34 (m, 2H), 7.88 (m, 2H), 8.39 (t, 1H, J = 5.93Hz), 12.91 (br, 1H)

10. Synthesis of Exemplified Compound 8 Exemplified Compound 8 was synthesized according to the synthesis method of Comparative Compound A using n-decanoic acid chloride instead of acetic anhydride (yield 62.7%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 0.85 (t, 3H, J = 7.06 Hz), 1.18-1.30 (m, 12H), 1.51 (m, 2H), 2.13 (t, 2H, J = 7.34Hz), 4.31 (d, 2H, J = 5.93Hz), 7.34 (d, 2H, J = 8.28Hz), 7.88 (d, 2H, J = 8.28Hz), 8.39 (t, 1H, J = 6.02Hz) ), 12.89 (br, 1H)

11. Synthesis of Exemplified Compound 9 Exemplified Compound 9 was synthesized according to the synthesis method of Comparative Compound A using lauric acid chloride instead of acetic anhydride (yield 87.8%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 0.85 (t, 3H, J = 7.06 Hz), 1.18-1.30 (m, 16H), 1.51 (m, 2H), 2.13 (t, 2H, J = 7.34Hz), 4.31 (d, 2H, J = 6.12Hz), 7.34 (d, 2H, J = 8.28Hz), 7.88 (d, 2H, J = 8.28Hz), 8.39 (t, 1H, J = 6.02Hz) ), 12.89 (br, 1H)

12 Synthesis of Exemplified Compound 10 Exemplified Compound 10 was synthesized according to the synthesis method of Comparative Compound A using myristic acid chloride instead of acetic anhydride (yield 78.1%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (Pyridine-d5, 600 MHz) 0.85 (t, 3H, J = 7.01 Hz), 1.16-1.30 (m, 18H), 1.39 (m, 2H), 1.87 (m, 2H), 2.50 (t , 2H, J = 7.34Hz), 4.78 (d, 2H, J = 5.83Hz), 7.61 (d, 2H, J = 8.28Hz), 8.42 (d, 2H, J = 8.09Hz), 9.15 (t, 1H , J = 5.65Hz)

13. Synthesis of Exemplified Compound 11 Exemplified Compound 11 was synthesized according to the synthesis method of Comparative Compound A using palmitic acid chloride instead of acetic anhydride (yield 50.9%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (Pyridine-d5, 600 MHz) 0.85 (t, 3H, J = 6.78 Hz), 1.15-1.32 (m, 22H), 1.39 (m, 2H), 1.87 (m, 2H), 2.49 (t , 2H, J = 7.34Hz), 4.77 (d, 2H, J = 5.36Hz), 7.61 (d, 2H, J = 7.53Hz), 8.41 (d, 2H, J = 7.91Hz), 9.14 (t, 1H , J = 5.74Hz)

14 Synthesis of Exemplified Compound 12 4- (Nt-butoxycarbonyl) aminomethylbenzoic acid methyl ester (1.33 g, 5 mmol) was dissolved in 4M-hydrochloric acid / dioxane solution (30 mL), stirred at room temperature for 1 hour, and then reduced in pressure. The solvent was distilled off under. The residue was dissolved in chloroform (20 mL), and stirred at 0 ° C., to which oleic acid (4.75 mmol), 1-Ethyl-3- (3-dimethylaminopropyl) carbodiimide Hydrochloride (EDC) (5 mmol), 1-hydroxy Benzotriazole (5 mmol) and triethylamine (5.5 mmol) were added and stirring was continued overnight from that temperature to room temperature. After completion of the reaction, a saturated aqueous solution of sodium hydrogen carbonate (30 mL) was added, and the mixture was extracted with chloroform (50 mL). The organic layer was washed with a saturated aqueous solution of sodium hydrogencarbonate (50 mL), 2N-sulfuric acid (50 mL) and saturated brine (50 mL), dried over magnesium sulfate and filtered, and then the solvent was distilled off under reduced pressure. The residue (2.09 g) was purified by silica gel column chromatography to obtain 4-oleoylaminomethylbenzoic acid methyl ester (0.60 g, yield 29.4%) as an intermediate. Next, the intermediate (0.56 g, 1.30 mmol) is dissolved in dioxane (20 mL), and an aqueous sodium hydroxide solution (2.60 mmol of sodium hydroxide, 10 mL of water) is added thereto, and refluxed at 100 ° C. overnight. did. After cooling to room temperature, 2N-sulfuric acid (10 mL) was added, the system was acidified, extracted with chloroform (50 mL), the organic layer was separated, and washed with saturated brine (30 mL) three times. After drying over magnesium sulfate and filtration, the solvent was distilled off under reduced pressure, and the residue (0.55 g) was purified by silica gel column chromatography to give Exemplary Compound 12 (0.40 g, yield 74.1 based on the intermediate). %). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (DMSO-d6, 600 MHz) δ 0.84 (t, 3H, J = 7.06 Hz), 1.17-1.33 (m, 20H), 1.51 (m, 2H), 1.98 (m, 4H), 2.13 (t, 2H, J = 7.534Hz), 4.31 (d, 2H, J = 5.93Hz), 5.29-5.35 (m, 2H), 7.33 (d, 2H, J = 8.09Hz), 7.88 (d, 2H, J = 8.38Hz), 8.39 (t, 1H, J = 6.02Hz), 12.93 (br, 1H)

15. Synthesis of Exemplified Compound 13 Tyramine hydrochloride (1.0 g, 5.76 mmol) was dissolved in chloroform (15 mL), and while stirring at 0 ° C., acetic anhydride (11.52 mmol, 1.10 mL), triethylamine (23.04 mmol) , 3.22 mL) was added and stirred overnight. After completion of the reaction, 2N-sulfuric acid (30 mL) was added to acidify the system, and the mixture was extracted with chloroform (50 mL). The separated organic layer was washed twice with a saturated aqueous solution of sodium hydrogencarbonate (50 mL), dried over magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue (0.97 g) was dissolved in methanol (15 mL), and an aqueous solution (15 mL) of sodium hydroxide (20 mmol, 0.8 g) was added and stirred overnight at room temperature. After completion of the reaction, 2N-sulfuric acid (30 mL) was added, the system was acidified, extracted with ethyl acetate (50 mL), and the solvent was evaporated under reduced pressure. The residue (0.78 g) was purified by silica gel column chromatography to give Exemplified Compound 13 (0.68 g, 66% yield). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (CD3OD, 600 MHz) δ 1.94 (s, 3H), 2.72 (t, 2H, J = 7.34 Hz), 3.36 (t, 2H, J = 7.34 Hz), 6.74 (m, 2H), 7.06 (m, 2H)

16. Synthesis of Exemplified Compound 14 Exemplified Compound 14 was synthesized according to the synthesis method of Exemplified Compound 13 using n-butanoic acid chloride instead of acetic anhydride (yield 28.6%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (CD3OD, 600 MHz) δ 0.95 (t, 3H, J = 7.44 Hz), 1.64 (m, 2H), 2.16 (t, 2H, J = 7.48 Hz), 2.72 (t, 2H, J = 7.62Hz), 3.38 (t, 2H, J = 7.62Hz), 6.74 (m, 2H), 7.06 (m, 2H)

17. Synthesis of Exemplified Compound 15 Exemplified Compound 15 was synthesized according to the synthesis method of Exemplified Compound 13 using benzoyl chloride instead of acetic anhydride (yield 23.0%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (CD3OD, 600 MHz) δ 2.85 (t, 2H, J = 7.62 Hz), 3.57 (t, 2H, J = 7.62 Hz), 6.76 (m, 2H), 7.11 (m, 2H), 7.48 (m, 2H), 7.56 (m, 1H), 7.80 (m, 2H)

18. Synthesis of Exemplified Compound 16 Exemplified Compound 16 was synthesized according to the synthesis method of Exemplified Compound 13 using 2-ethylhexanoic acid chloride instead of acetic anhydride (yield 24.3%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (CD3OD, 600 MHz) δ 0.87 (t, 3H, J = 7.43 Hz), 0.92 (t, 3H, J = 7.34 Hz), 1.21 (m, 2H), 1.26-1.48 (m, 4H ), 1.55 (m, 2H), 2.07 (m, 1H), 2.74 (t, 2H, J = 7.15Hz), 3.42 (m, 2H,), 6.74 (m, 2H), 7.08 (m, 2H)

19. Synthesis of Exemplified Compound 17 Exemplified Compound 17 was synthesized according to the synthesis method of Exemplified Compound 13 using 3-phenylpropanoic acid chloride instead of acetic anhydride (yield 25.8%). The structure of the obtained compound was confirmed by ¹ H-NMR.
¹ H-NMR: (CD3OD, 600 MHz) 2.47 (t, 2H, J = 7.53 Hz), 2.64 (t, 2H, J = 7.43 Hz), 2.91 (t, 2H, J = 7.53 Hz), 3.33 (t, 2H, J = 7.43Hz), 6.72 (m, 2H,), 6.98 (m, 2H), 7.19-7.24 (m, 3H), 7.29 (m, 2H)

Example Measurement of Dopa Oxidase Activity In a 96-well plate, 100 μL of human neonatal foreskin-derived melanocytes were seeded in each hole to a cell density of 1 × 10 ⁴ cells / well. The medium used was medium 254 supplemented with HMGS (Human Melanocyte Growth Supplement) excluding PMA (both manufactured by Cascade Biologics).
After culturing for 24 hours, melanocyte activator endothelin-1 (ET-1), stem cell growth factor (SCF), α melanocyte stimulating hormone (α-MSH), histamine and prostaglandin E ₂ (PGE ₂ ), respectively, The final concentration in the medium was 10 × 10 ⁻⁷ mol / m ³ .

Exemplified compounds 1 to 17 and comparative compounds A and B produced in the above synthesis examples were added as samples so that the final concentrations shown in Table 1 were obtained. As a reference example, a sample was prepared by adding known kojic acid as a whitening component having high inhibitory activity against tyrosinase to a final concentration of 0.5 mM.
Finally, the amount of the medium was 200 μL / well, and the cells were cultured for 3 days at 37 ° C. and 5% CO ₂ .

In addition, the following additives are also added to the medium.
bFGF (basic fibroblast growth factor) 3 ng / mL
BPE (bovine pituitary extract) 0.2% by volume
FBS (fetal bovine serum) 0.5% by volume
Hydrocortisone 5 × 10 ^-4 mol / m ³
Insulin 5 μg / mL
Transferrin 5 μg / mL
Heparin 5μg / mL

After completion of the culture, 20 μL of Alamar Blue (Alamar Blue, trade name, manufactured by Invitrogen) reagent is added to each well, and after 2-3 hours of incubation, the fluorescence intensity of the medium (excitation wavelength: 544 nm, fluorescence wavelength: 590 nm) is measured. The cell proliferation activity was measured.
Melanocytes whose cell proliferation activity was measured were washed with Phosphate-buffered saline (PBS) from which Ca ²⁺ and Mg ²⁺ were removed, and extracted buffer (0.1 M Tris-HCL (pH 7.2), 1% Nonidet P-40 , 0.01% SDS, 100 μM PMSF (phenylmethylsulfonyl fluoride), 1 μg / mL aprotinin) at 20 μL / well, Assay buffer (100 mM Sodium phosphate-buffered (pH 7.1) containing 4% dimethylformamide) at 20 μL / Well was added, the cells were solubilized at 4 ° C. for 3 hours, and the dopa oxidase activity was measured. The dopa oxidase activity was measured as follows with reference to the MBTH method (see, for example, Winder AJ, Harris H., Eur. J. Biochem., 198, 317-326, 1991).
In each well of the solubilized cell solution, Assay buffer is 80 μL / well, 20.7 mM MBTH (3-methyl-2-benzothiazolinone hydrazone) solution is 60 μL, and 5 mM L-dopa (L-dihydroxyphenylalanine) is used as a substrate. After adding 40 μL of each solution and reacting at 37 ° C. for 30 to 60 minutes, the color reaction was measured by absorbance at 490 nm.

The results of cell proliferation activity measurement and dopa oxidase activity measurement are shown in Table 1. The cell proliferation activity values in Table 1 are shown as relative values to the fluorescence intensity when each sample was not added (control). Moreover, the dopa oxidase activity inhibition rate was computed by the following formula.

Dopa oxidase activity inhibition rate (%) = 100 − {(absorbance of system to which sample was added) / (absorbance of system to which sample was not added (control))} × 100

As is clear from the results in Table 1, in the system in which Exemplified Compounds 1 to 17 were added as samples, the dopa oxidase activity was greatly reduced compared to the control, and the dopa oxidase activity inhibition rate was high. On the other hand, in the system to which Comparative Compound A and Comparative Compound B were added, the dopa oxidase activity was larger or similar to that of the control, and the dopa oxidase activity was not inhibited at all. From these results, it is understood that the compound represented by the general formula (1) has an excellent dopa oxidase activity inhibitory action.
As described above, the dopa oxidase activity is used as an indicator of the enzyme activity of tyrosinase relating to melanin biosynthesis. Kojic acid shown as a reference example is a known whitening component having high inhibitory activity against tyrosinase. It can be seen that the compound represented by the general formula (1) can effectively suppress the dopa oxidase activity even at a relatively low concentration compared with kojic acid. Therefore, it turns out that the compound represented by the said General formula (1) can suppress melanin production by inhibiting dopa oxidase activity effectively, and is useful as a whitening component.

(Prescription example)
Using the extract obtained in the above production example as an active ingredient, lotions, emulsions, serums, creams and packs having the compositions shown below were prepared by conventional methods.

1. Preparation of lotion (composition) (Formulation: mass%)
Exemplary Compound 4 0.05
1,3-butylene glycol 8.0
Glycerin 5.0
Ethanol 3.0
Chamomile extract 3.0
Kyokyo Extract 1.0
Clove extract 1.0
Xanthan gum 0.1
Hyaluronic acid 0.1
Disodium phosphate 0.1
Sodium dihydrogen phosphate 0.1
Ascorbic acid 2-glucoside 2.0
Purified water balance Perfume appropriate amount Preservative appropriate amount

2. Preparation of lotion (composition)
Exemplary Compound 13 0.1
1,3-butylene glycol 5.0
Glycerin 3.0
Propylene glycol 3.0
Ethanol 3.0
Chamomile extract 2.0
Xanthan gum 0.1
Hyaluronic acid 0.1
Disodium phosphate 0.1
Sodium dihydrogen phosphate 0.1
Purified water balance Perfume appropriate amount Preservative appropriate amount

3. Preparation of milky lotion (composition) (formulation: mass%)
Exemplary Compound 4 0.05
Chamomile extract 1.0
Kyokyo Extract 1.0
Altea extract 2.0
Squalane 3.0
Olive oil 3.0
Glycerin 5.0
Polyoxyethylene hydrogenated castor oil (number of moles of ethylene oxide added: 40) 1.0
Carboxyvinyl polymer 0.2
Potassium hydroxide 0.1
Xanthan gum 0.2
Edetate disodium 0.02
Purified water balance Preservative

4). Preparation of serum (composition) (Composition: mass%)
Exemplary Compound 5 0.05
Chamomile extract 1.0
Kyokyo Extract 1.0
Clove extract 1.0
Carboxyvinyl polymer 0.2
Acrylic acid / alkyl methacrylate copolymer 0.2
Potassium hydroxide 0.2
Xanthan gum 0.1
Hyaluronic acid 0.2
Sodium citrate 0.15
Citric acid 0.03
Glycerin 10.0
1,3-butylene glycol 5.0
Edetate dinatrim 0.05
Purified water balance Preservatives proper amount Fragrance proper amount

5. Preparation of serum (composition) (Composition: mass%)
Exemplary Compound 14 0.1
Chamomile extract 1.0
Clove extract 1.0
Kyokyo Extract 1.0
Xanthan gum 0.2
Carboxymethylcellulose 0.2
Carboxyvinyl polymer 0.2
Potassium hydroxide 0.1
Citric acid 0.03
Sodium citrate 0.15
Glycerin 5.0
Propylene glycol 3.0
Polyethylene glycol (molecular weight 1500) 1.0
Polyethylene glycol monostearate 0.5
Purified water balance Preservatives proper amount Fragrance proper amount

6). Preparation (composition) of cream (formulation: mass%)
Exemplary Compound 6 0.1
Chamomile extract 2.0
Kyokyo Extract 2.0
Clove extract 2.0
Methylpolysiloxane 3.0
Squalane 2.0
Neopentyl glycol dicaprate 3.0
Stearyl alcohol 1.5
Cetanol 1.0
Polyoxyethylene hydrogenated castor oil (number of moles of ethylene oxide added: 60) 0.5
Acrylic acid / alkyl methacrylate copolymer 0.3
Potassium hydroxide 0.15
Xanthan gum 0.1
Edetate disodium 0.05
Purified water balance Preservatives proper amount Fragrance proper amount

7). Preparation (composition) of cream (formulation: mass%)
Exemplary Compound 16 0.01
Chamomile extract 3.0
Kyokyo Extract 3.0
Polyoxyethylene alkyl ether modified silicone 3.0
Methylpolysiloxane 5.0
Decamethylcyclopentasiloxane 10
Squalane 5.0
Magnesium sulfate 0.5
Glycerin 5.0
1,3-butylene glycol 5.0
Purified water balance Preservatives proper amount Fragrance proper amount

8). Preparation of pack (composition) (Formulation: mass%)
Exemplary Compound 7 0.01
Chamomile extract 2.0
Kyokyo Extract 1.0
Dipropylene glycol 3.0
Polyethylene glycol (average molecular weight 1500) 2.0
1,3-butylene glycol 5.0
Glycerin 5.0
Sodium citrate 0.5
Polyvinyl alcohol 10
Lactic acid 0.3
Polyoxyethylene decyl tetredecyl ether 0.5
Purified water balance Preservatives proper amount Fragrance proper amount

Claims (2)

A dopa oxidase activity inhibitor comprising a compound represented by the following general formula (1) or a salt thereof as an active ingredient.

(In the formula, X represents a hydroxyl group or a carboxyl group. R represents an alkyl group, an alkenyl group, an aryl group or an aralkyl group, and these groups may be substituted. However, when X is a carboxyl group, R represents Group having 4 or more carbon atoms in total. N represents 1 or 2.) A whitening agent containing a compound represented by the following general formula (1) or a salt thereof as an active ingredient.

(In the formula, X represents a hydroxyl group or a carboxyl group. R represents an alkyl group, an alkenyl group, an aryl group or an aralkyl group, and these groups may be substituted. However, when X is a carboxyl group, R represents Group having 4 or more carbon atoms in total. N represents 1 or 2.)