KR102166292B1 - Skin whitening composition comprising quinine based compounds as effective ingredients and method for screening the same compounds - Google Patents

Abstract

The present invention relates to a composition for skin whitening comprising a quinone compound represented by the following structural formula 1 as an active ingredient. Thereby, such a composition contains a quinone-based compound screened from a quinone-based compound library derived from a natural product, and exhibits an inhibitory effect on melanogenesis through the Wnt/β-catenin signaling pathway. It shows very excellent effects in the treatment and prevention of skin whitening and hyperpigmentation diseases.
[Structural Formula 1]

Description

Skin whitening composition containing a quinone compound as an active ingredient, and a screening method for the compound {SKIN WHITENING COMPOSITION COMPRISING QUININE BASED COMPOUNDS AS EFFECTIVE INGREDIENTS AND METHOD FOR SCREENING THE SAME COMPOUNDS}

The present invention relates to a composition for skin whitening comprising a quinone compound as an active ingredient and a screening method for the compound, and more particularly, to a skin whitening and hyperpigmentation by inhibiting melanogenesis by including a quinone compound as an active ingredient. It relates to a method for screening a compound having a function of inhibiting melanogenesis from a cosmetic composition, pharmaceutical composition and food composition effective against diseases, and a quinone compound library.

Melanin, which determines the color of a person's skin, is made in skin cells called melanocytes and moves to epidermal cells called keratinocytes. Here, melanin plays an important role in protecting genes from ultraviolet rays and protecting proteins in cells by removing free radicals. There is no enzyme that degrades melanin in the body, but when keratinocytes (keratinocytes) fall off the epidermis, they are removed by falling off the skin.

When more melanin is produced than necessary, it causes hyperpigmentation such as spots, freckles, and spots, leading to unfavorable results in terms of beauty or health. In addition, as the number of people who enjoy activities outside due to the increase of the leisure population increases, the demand to prevent melanin pigmentation caused by ultraviolet rays increases. The development of whitening agents to date has mainly consisted of finding substances that reduce the amount of melanin by inhibiting the activity of tyrosinase, an enzyme that plays an essential and most important role in melanin biosynthesis. The whitening agent developed in this way includes hydro-quinone, thiol, kojic acid, arbutin, vitamin C, etc., but does not have satisfactory whitening effect. The attempt to find and develop a material that replaces it by causing or causing it occupies a key part of material research.

Melanin synthesis is regulated by a complex network of systematic hormones and local factors, and occurs as a result of the upregulation of microphthalmia-associated transcription factor (MITF), a major regulator of melanin biosynthesis. Activates the expression of synase and tyrosinase-related proteins (Eur. Cell Mater. 28: 269-286; J. Biol. Chem. 284: 32533-32543; Cell Death Dis. 5:e1187; World J. Exp. Med. 2: 78-855; Mayo Clin. Proc. 77:453-468; Eur. J. Endocrinol. 165: 1-10.)

The Wnt/β-catenin pathway regulates various biological processes including the development of neural tube-derived melanocytes, and activation of the pathway induces stabilization of intracellular β-catenin that regulates the target gene of the pathway. β-catenin not only forms a complex with lymphocyte enhancer factor-1, which activates MITF gene expression, but also upregulates MITF-dependent genes through direct binding with MITF protein. In addition, mice lacking Wnt1- and Wnt3- are characterized by a lack of pigment cells due to the failure of initial proliferation of neural tube cells.

Korean Patent Publication No. 10-0716891

An object of the present invention is to provide a skin whitening cosmetic composition capable of exhibiting a skin whitening effect by inhibiting melanin synthesis.

Another object of the present invention is to provide a food composition for skin whitening that can exhibit a skin whitening effect by inhibiting melanin synthesis.

Another object of the present invention is to provide a cosmetic composition for preventing or treating hyperpigmentation disease, which can exhibit a skin whitening effect by inhibiting melanin synthesis.

Another object of the present invention is to provide a method for screening a natural product-derived compound having efficacy in improving skin whitening and hyperpigmentation diseases by weakening the Wnt/β-catenin pathway in the direction of reducing melanin components by inhibiting the differentiation of melanocytes Have.

According to an aspect of the present invention,

A cosmetic composition for skin whitening comprising a quinone-based compound represented by the following structural formula 1 as an active ingredient is provided.

[Structural Formula 1]

In structural formula 1,

n is a repeating number that is an integer selected from 1 to 10,

R ¹ is a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, a substituted or unsubstituted C6 to C30 aryl group, Or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, A substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, It is a substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group.

Preferably, in the structural formula 1,

n is a repeating number that is an integer selected from 1 to 3,

R ¹ is a C1 to C10 alkyl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom or a C1 to C10 alkyl group,

R ⁴ to R ⁷ may be the same as or different from each other, and may each independently be a hydrogen atom or a C1 to C10 alkyl group.

More preferably, the compound represented by the structural formula 1 may be lucidin-3-methyl ether.

The cosmetic composition for skin whitening may have a function of inhibiting the synthesis of β-catenin protein.

The cosmetic composition for skin whitening may have a function of inhibiting melanocyte differentiation.

According to another aspect of the present invention,

There is provided a pharmaceutical composition for preventing or treating hyperpigmentation diseases comprising a quinone-based compound represented by the following structural formula 1 as an active ingredient.

[Structural Formula 1]

In structural formula 1,

n is a repeating number that is an integer selected from 1 to 10,

R ¹ is a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, a substituted or unsubstituted C6 to C30 aryl group, Or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, A substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, It is a substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group.

Preferably, in the structural formula 1,

n is a repeating number that is an integer selected from 1 to 3,

R ¹ is a C1 to C10 alkyl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom or a C1 to C10 alkyl group,

R ⁴ to R ⁷ may be the same as or different from each other, and may each independently be a hydrogen atom or a C1 to C10 alkyl group.

More preferably,

The compound represented by Structural Formula 1 may be lucidin-3-methyl ether.

According to another aspect of the present invention,

There is provided a food composition for preventing or improving hyperpigmentation disease comprising a quinone-based compound represented by the following structural formula 1 as an active ingredient.

[Structural Formula 1]

In structural formula 1,

n is a repeating number that is an integer selected from 1 to 10,

R ¹ is a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, a substituted or unsubstituted C6 to C30 aryl group, Or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, A substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, It is a substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group.

Preferably, in the structural formula 1,

n is a repeating number that is an integer selected from 1 to 3,

R ¹ is a C1 to C10 alkyl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom or a C1 to C10 alkyl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently may be a hydrogen atom or a C1 to C10 alkyl group.

More preferably, the compound represented by the structural formula 1 may be lucidin-3-methyl ether.

According to another aspect of the present invention,

A method for screening whitening active compounds using a Wnt/β-catenin signal from a quinone compound library is provided.

The whitening active compound may inhibit Wnt/β-catenin signal.

Inhibiting the Wnt/β-catenin signal may be caused by a decrease in the level of β-catenin protein.

For the screening method, a HEK293-FL (HEK293-Firefly luciferase) reporter cell, which is a HEK293 cell line into which a human Frizzled-1 expression plasmid and a β-catenin/Tcf-dependent luciferase reporter plasmid are introduced, can be used.

The HEK293-FL reporter cells may be cultured in a Wnt3a-conditioned medium with a compound of a quinone-based compound library.

HEK293-FL reporter cells may be cultured in DMEM medium before being cultured in a medium under Wnt3a conditions with the compound of the quinone-based compound library.

The medium under the Wnt3a condition may be a DMEM medium under the Wnt3a condition (Wnt3a-CM).

Cosmetic compositions, food compositions, and pharmaceutical compositions comprising the quinone-based compound screened using the Wnt/β-catenin signal of the present invention as an active ingredient use the Wnt/β-catenin signaling pathway. As a result, it exhibits a melanin production inhibitory effect, and as a result, it can exhibit very excellent effects in the treatment and prevention of diseases such as skin whitening and hyperpigmentation.

1 is a result of confirming the effect of inhibiting Wnt/β-catenin signal according to the concentration of lucidin-3-methyl ether of Experimental Example 1.
2 is a result of analysis of the effect of the concentration of lucidin-3-methyl ether on the level of β-catenin protein in the cytoplasm of Experimental Example 2.
3 is a result of analyzing the effect of the concentration of lucidin-3-methyl ether on the level of β-catenin mRNA in the cytoplasm of Experimental Example 3.
4 is a result of analyzing the effect on the melanin formation process according to the concentration of lucidin-3-methyl ether of Experimental Example 4.

Since the present invention can apply various transformations and have various embodiments, specific embodiments are illustrated in the drawings and will be described in detail in the detailed description. However, this is not intended to limit the present invention to a specific embodiment, it is to be understood to include all conversions, equivalents, and substitutes included in the spirit and scope of the present invention. In describing the present invention, when it is determined that a detailed description of a related known technology may obscure the subject matter of the present invention, a detailed description thereof will be omitted.

The terms used in the present application are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present application, terms such as "comprise" or "have" are intended to designate the existence of features, numbers, steps, elements, or combinations thereof described in the specification, and one or more other features, numbers, and steps It is to be understood that the possibility of the presence or addition of, components, or combinations thereof, is not excluded in advance.

In the present invention, "substituted" means that at least one hydrogen atom is deuterium, C1 to C30 alkyl group, C3 to C30 cycloalkyl group, C2 to C30 heterocycloalkyl group, C1 to C30 halogenated alkyl group, C6 to C30 aryl group, C1 to C30 heteroaryl Group, C1 to C30 alkoxy group, C3 to C30 cycloalkoxy group, C1 to C30 heterocycloalkoxy group, C2 to C30 alkenyl group, C2 to C30 alkynyl group, C6 to C30 aryloxy group, C1 to C30 heteroaryloxy group, Silyloxy group (-OSiH ₃ ), -OSiR ¹ H ₂ (R ¹ is a C1 to C30 alkyl group or C6 to C30 aryl group), -OSiR ¹ R ² H (R ¹ and R ² are each independently a C1 to C30 alkyl group Or C6 to C30 aryl group), -OSiR ¹ R ² R ³ , (R ¹ , R ² , and R ³ are each independently a C1 to C30 alkyl group or a C6 to C30 aryl group), C1 to C30 acyl group, C2 to C30 acyloxy group, C2 to C30 heteroaryloxy group, C1 to C30 sulfonyl group, C1 to C30 alkylthiol group, C3 to C30 cycloalkylthiol group, C1 to C30 heterocycloalkylthiol group, C6 to C30 arylthiol group, C1 to C30 heteroarylthiol group, C1 to C30 phosphate amide group, silyl group (SiR ¹ R ² R ³ ₎ (R ¹ , R ² , and R ³ are each independently a hydrogen atom, C1 to C30 alkyl group or C6 to C30 Aryl group), amine group (-NRR') (wherein R and R'are each independently a hydrogen atom, a substituent selected from the group consisting of a C1 to C30 alkyl group, and a C6 to C30 aryl group), carboxyl group, halogen It means substituted with a substituent selected from the group consisting of a group, a cyano group, a nitro group, an azo group, and a hydroxy group.

In addition, two adjacent substituents among the substituents may be fused to form a saturated or unsaturated ring.

In addition, the range of carbon number of the alkyl group or aryl group in the "substituted or unsubstituted C1 to C30 alkyl group" or "substituted or unsubstituted C6 to C30 aryl group" etc. is unsubstituted without considering the portion where the substituent is substituted It refers to the total number of carbon atoms constituting the alkyl moiety or the aryl moiety when viewed as being formed. For example, a phenyl group in which a butyl group is substituted in the para position corresponds to an aryl group having 6 carbon atoms substituted with a butyl group having 4 carbon atoms.

In the present specification, unless otherwise defined, "hetero" means that one functional group contains 1 to 4 heteroatoms selected from the group consisting of N, O, S and P, and the rest are carbon.

In the present specification, "hydrogen" refers to singlet hydrogen, dihydrogen, or tritium unless otherwise defined.

In the present specification, "alkyl (alkyl) group" refers to an aliphatic hydrocarbon group unless otherwise defined.

The alkyl group may be a "saturated alkyl group" that does not contain any double bonds or triple bonds.

The alkyl group may be an "unsaturated alkyl group" including at least one double bond or triple bond.

The alkyl group, whether saturated or unsaturated, can be branched, straight-chain or cyclic.

The alkyl group may be a C1 to C30 alkyl group. More specifically, it may be a C1 to C20 alkyl group, a C1 to C10 alkyl group, or a C1 to C6 alkyl group.

For example, a C1 to C4 alkyl group has 1 to 4 carbon atoms in the alkyl chain, i.e., the alkyl chain is methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl and t-butyl. It indicates selected from the group consisting of.

For specific examples, the alkyl group is a methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, t-butyl group, pentyl group, hexyl group, ethenyl group, propenyl group, butenyl group, cyclopropyl group, cyclo It means a butyl group, a cyclopentyl group, a cyclohexyl group, etc.

“Cycloalkyl groups” include monocyclic or fused ring polycyclic (ie, rings that share adjacent pairs of carbon atoms) functional groups.

"Heterocycloalkyl group" means that the cycloalkyl group contains 1 to 4 heteroatoms selected from the group consisting of N, O, S, and P, and the remainder is carbon. When the heterocycloalkyl group is a fused ring, at least one ring among the fused rings may contain 1 to 4 hetero atoms.

“Aryl groups” include monocyclic or fused ring polycyclic (ie, rings that share adjacent pairs of carbon atoms) functional groups.

"Heteroaryl group" means that the aryl group contains 1 to 4 heteroatoms selected from the group consisting of N, O, S and P, and the remainder is carbon. When the heteroaryl group is a fused ring, at least one ring among the fused rings may contain 1 to 4 hetero atoms.

In the aryl group and the heteroaryl group, the number of ring atoms is the sum of the number of carbon atoms and the number of non-carbon atoms.

Hereinafter, the cosmetic composition for skin whitening of the present invention will be described.

The cosmetic composition for skin whitening of the present invention comprises a quinone-based compound represented by the following structural formula 1 as an active ingredient.

[Structural Formula 1]

In structural formula 1,

n is a repeating number that is an integer selected from 1 to 10,

R ¹ is a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, a substituted or unsubstituted C6 to C30 aryl group, Or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, A substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, It is a substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group.

Preferably, in the structural formula 1,

n is a repeating number that is an integer selected from 1 to 3,

R ¹ is a C1 to C10 alkyl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom or a C1 to C10 alkyl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently may be a hydrogen atom or a C1 to C10 alkyl group.

More preferably, the compound represented by the structural formula 1 may be lucidin-3-methyl ether.

The cosmetic composition for skin whitening may have a function of inhibiting the synthesis of β-catenin protein.

The cosmetic composition for skin whitening may have a function of inhibiting melanocyte differentiation.

The inhibition of melanocyte differentiation may be achieved through an action of inhibiting the expression of microphthalmia associated transcription factor (MITF).

Ingredients included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions in addition to the above active ingredients as an active ingredient, for example, conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, fragrances, And a carrier. The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing containing surfactant, oil , Powder foundation, emulsion foundation, wax foundation may be formulated, but is not limited thereto. In more detail, it may be prepared in formulations such as cleansing, lotion, cream, massage cream, eye cream, essence, gel, pack, spray, powder, sun cream for face wash.

When the formulation of the present invention is a paste, cream or gel, vegetable oil, wax, starch, cellulose, silicone, bentonite, silica, zinc oxide, etc. may be used as a carrier component. When the formulation is powder, lactose and silica as carrier components , Aluminum hydroxide, calcium silicate, and the like powder may be used. When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, such as water, ethanol, glycerin, ethyl carbonate, ethyl acetate, propylene glycol, butylene glycol, 1 ,2-hexanediol, glycerol aliphatic ester, and sorbitan fatty acid ester.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, and the like may be used. In addition, when the formulation is a surfactant-containing cleansing, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, methyltaurate, sarcosinate, alkylamidobetaine, aliphatic alcohol, fatty acid glycerol Ride, vegetable oil or ethoxylated glycerol fatty acid esters and the like may be used.

Hereinafter, a pharmaceutical composition for preventing or treating diseases of hyperpigmentation of the present invention will be described.

As used herein, the term "hyperpigmentation" refers to darkening or darkening in a specific area of the skin or nail compared to other areas due to an excessive increase in melanin.

The hyperpigmentation disease may be melasma, freckles, senile pigment spots, solar lentigines, and the like, but the scope of the present invention is not limited thereto.

The pharmaceutical composition for preventing or treating hyperpigmentation diseases of the present invention comprises a quinone-based compound represented by the following structural formula 1 as an active ingredient.

[Structural Formula 1]

In structural formula 1,

n is a repeating number that is an integer selected from 1 to 10,

R ¹ is a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, a substituted or unsubstituted C6 to C30 aryl group, Or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, A substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, It is a substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group.

Preferably, in the structural formula 1,

n is a repeating number that is an integer selected from 1 to 3,

R ¹ is a C1 to C10 alkyl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom or a C1 to C10 alkyl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently may be a hydrogen atom or a C1 to C10 alkyl group.

More preferably, the compound represented by the structural formula 1 may be lucidin-3-methyl ether.

The pharmaceutical composition for preventing or treating hyperpigmentation disease may have a function of inhibiting the synthesis of β-catenin protein.

The pharmaceutical composition for preventing or treating hyperpigmentation disease may have a function of inhibiting melanocyte differentiation.

The inhibition of melanocyte differentiation may be achieved through an action of inhibiting the expression of microphthalmia associated transcription factor (MITF).

In addition to the active ingredient, the pharmaceutical composition of the present invention may be prepared using a physiologically acceptable auxiliary agent, and the auxiliary agent includes an excipient, a disintegrant, a sweetener, a binder, a coating agent, an expanding agent, a lubricant, and a lubricant. Alternatively, flavoring agents or the like can be used.

For administration, the pharmaceutical composition may contain one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.

The formulation form of the pharmaceutical composition may be a liquid, syrup, juice, suspension, emulsion, drop, or the like. For example, also, if necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like. It is preferable that the pharmaceutical composition of the present invention is administered by transdermal administration.

Hereinafter, a food composition for preventing or improving the hyperpigmentation disease of the present invention will be described.

The food composition for preventing or improving hyperpigmentation disease of the present invention comprises a quinone compound represented by the following structural formula 1 as an active ingredient.

[Structural Formula 1]

In structural formula 1,

n is a repeating number that is an integer selected from 1 to 10,

R ¹ is a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, a substituted or unsubstituted C6 to C30 aryl group, Or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, A substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently a hydrogen atom, a substituted or unsubstituted C1 to C30 alkyl group, a substituted or unsubstituted C3 to C30 cycloalkyl group, a substituted or unsubstituted C1 to C30 heterocycloalkyl group, It is a substituted or unsubstituted C6 to C30 aryl group, or a substituted or unsubstituted C1 to C30 heteroaryl group.

Preferably, in the structural formula 1,

n is a repeating number that is an integer selected from 1 to 3,

R ¹ is a C1 to C10 alkyl group,

R ² and R ³ are the same as or different from each other, and each independently a hydrogen atom or a C1 to C10 alkyl group,

R ⁴ to R ⁷ are the same as or different from each other, and each independently may be a hydrogen atom or a C1 to C10 alkyl group.

More preferably, the compound represented by the structural formula 1 may be lucidin-3-methyl ether.

The food composition for preventing or improving the hyperpigmentation disease may have a function of inhibiting the synthesis of β-catenin protein.

The food composition for preventing or improving the hyperpigmentation disease may have a function of inhibiting melanocyte differentiation.

The inhibition of melanocyte differentiation may be achieved through an action of inhibiting the expression of microphthalmia associated transcription factor (MITF).

The food composition of the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the food composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gums, ice creams, vitamin complexes, health supplements. And foods.

The food composition of the present invention may include not only the above active ingredients, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, in addition to the active ingredients of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be further included.

Hereinafter, a method for screening the whitening active compound of the present invention will be described.

The screening method of the whitening active compound of the present invention may be performed including the step of treating reporter cells expressing Wnt/β-catenin signal in a quinone-based compound library.

The whitening active compound is characterized in that it inhibits the Wnt/β-catenin signal.

Inhibiting the Wnt/β-catenin signal may be caused by a decrease in the level of β-catenin protein.

The screening method may be performed using a human Frizzled-1 expression plasmid and a β-catenin/Tcf-dependent luciferase reporter plasmid, a HEK293 cell line, HEK293-FL (HEK293-Firefly luciferase) reporter cells. .

HEK293-FL reporter cells can be cultured in DMEM medium before being cultured in a medium under Wnt3a conditions with the compound of the quinone-based compound library.

Thereafter, the HEK293-FL reporter cells may be cultured in a Wnt3a-conditioned medium with a compound of a quinone-based compound library.

The medium under the Wnt3a condition is preferably a DMEM medium under the Wnt3a condition (Wnt3a-CM).

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention, but the following examples are only illustrative of the present invention, and it is obvious to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It is natural that such modifications and modifications fall within the appended claims.

[Example]

Example One: Wnt /β- catenin Screening for compounds that inhibit signal

Human Frizzled-1 (hFz-1) expression plasmid and β-catenin/Tcf-dependent luciferase reporter plasmid are stable in Kookmin University's natural product-based quinone-based compound library to screen for compounds that inhibit Wnt/β-catenin signaling. HEK293-FL reporter cells introduced as were utilized (Mol. Pharmacol. 70(3): 960-966).

HEK293-FL reporter cells were dispensed in duplicate at a density of 2.5×10 ⁴ to each well of a 96-well plate, and 10% FBS, 120 mg/ml penicillin, and 200 mg/ml streptomycin were added in DMEM medium Incubated for 24 hours.

Cultured reporter cells were cultured in Wnt3a-CM (Wnt3a-conditioned medium (Wnt3a, 10% FBS, 120 mg/ml penicillin, 200 mg/ml streptomycin added DMEM medium) in Kookmin University's natural product-based quinone compound library The compounds were added and incubated for an additional 15 hours to a final concentration of 10 μM.

The additionally cultured reporter cells were measured for FL (firefly luciferase) activity and cell viability according to the manufacturer's instructions using the Dual-Luciferase Reporter Assay System (Promega) and CellTiter-Glo assay kit (Promega), respectively. FL activity in each well was normalized according to cell viability to evaluate the specific effect on the Wnt/β-catenin signal of the screened compound. Through this screening process, the compound lucidin-3-methyl ether was identified as the most probable inhibitor capable of inhibiting the increased FL (Firefly luciferase) activity by Wnt3a-CM. , The formula of lucidin-3-methyl ether is represented by the following formula (1).

[Formula 1]

[Experimental Example]

Experimental example One: Wnt /β- catenin Check signal suppression effect

Lucidin-3-methyl ether screened according to Example 1 was added to HEK293-FL reporter cells at 0, 7.5, 15, and 30 μM, respectively, and cultured for 15 hours using Wnt3a-CM, followed by Firefly luciferase. The activity was investigated, and the experiment was repeated 3 times, and the average value is shown in FIG. 1.

According to this, it was confirmed that lucidin-3-methyl ether significantly inhibited β-catenin transcription (CRT, Wnt3a-CM-induced β-catenin response transcription) induced by Wnt3a-CM as the dose increased. In contrast, when lucidin-3-methyl ether was treated in HEK293 control cells (introduced with hFz-1 and TOPFlash expression plasmid) under the same conditions, a negative control having a mutated β-catenin/Tcf-binding element It was found that it did not affect the activity of the reporter FOPFlash. Through this, it was confirmed that lucidin-3-methyl ether is a specific inhibitor of Wnt/β-catenin signal (antagonist).

Experimental example 2: β- in the cytoplasm catenin Analysis of the effect on protein levels

In the Wnt/β-catenin pathway, lucidin-3-methyl ether is added to the cytoplasmic β-catenin levels that regulate β-catenin transcription (CRT, Wnt3a-CM-induced β-catenin response transcription) induced by Wnt3a-CM. Western blotting was performed to confirm the effect.

For this purpose, HEK 293 cells were dispensed 6×10 ⁵ into a 60 mm culture dish and cultured in a culture medium for 24 hours. Thereafter, HEK293 cells were treated with lucidin-3-methyl ether and further incubated with Wnt3a-CM for 15 hours. Only the cytoplasmic fraction of the cells was extracted and subjected to SDS-PAGE followed by electrophoresis with a nitrocellulose membrane (Bio-Rad, Hercules, CA, USA). The nitrocellulose membrane was blocked with 5% fat-free milk and reacted with an anti-β-catenin antibody (BD transduction laboratories, Lexington, KY, USA) and an anti-β-actin antibody (Cell Signaling Technology, Denvers, MA, USA). . Thereafter, the nitrocellulose membrane was reacted with an anti-mouse IgG fused with HRP (horseradish-peroxidase) (Santa Cruz Biotechnology), and visualized using ECL (Santa Cruz Biotechnology), and the results are shown in FIG. 2. According to this, it was confirmed that the level of β-catenin in the cytoplasm was decreased depending on the concentration of lucidin-3-methyl ether.

Experimental example 3: In the cytoplasm β- catenin mRNA Analysis of impact on numbers

To confirm the effect of lucidin-3-methyl ether on the mRNA of β-catenin, total RNA was isolated according to the manufacturer's method using TRIzol reagent (Invitrogen), and cDNA synthesis, reverse transcription reaction, and PCR were used. The process proceeded. The amplified DNA was isolated on a 2% agarose gel and observed by EtBr staining.

According to this, it was confirmed that the level of β-catenin in the cytoplasm is a constant level regardless of the concentration of lucidin-3-methyl ether. It was found that Wnt/β-catenin signaling was suppressed by reducing catenin protein levels.

Experimental example 4: Analysis of the effect on the melanogenesis process

Since Wnt/β-catenin signals have been reported to regulate melanocyte differentiation, whether lucidin-3-methyl ether reduces melanin accumulation in B16 mouse melanoma cells was observed by absorbance analysis. B16 mouse melanoma cells was one to 8 × 10 to 60 mm tissue culture dish and adhered overnight ^four frequency divider, using a Lucy-3-methyl ether is added to the Wnt3a-CM and incubated for 4 days. The cells were washed 3 times with refrigerated PBS, and RIPA buffer [50 mM Tris-Cl (pH7.4)], 150 mM NaCl, 1% NP-40, 0.2% sodium dodecyl sulfate [(SDS), 5 mM NaF] After dissolving with, it was centrifuged for 10 minutes at 13,000 rpm (LaboGene). The supernatant was used for protein concentration analysis and western blotting, and the pellet was dissolved in 200µl of 1M NaOH and reacted at 60°C for 2 hours. A target curve was prepared using synthetic melanin (0-250 μg/ml), and absorbance was measured by spectrophotometry at 405 nm using a reader. Melanin production was calculated by normalizing the total amount of melanin to the protein concentration (melanin (µg)/protein (mg)), and the results are shown in FIG. 4. According to this, when Wnt3a-CM was treated in B16 mouse melanoma cells, melanin production increased, but when lucidin-3-methyl ether was treated, melanin accumulation was decreased in a concentration-dependent manner.

In the above, embodiments of the present invention have been described, but those of ordinary skill in the art will add, change, delete or add components within the scope not departing from the spirit of the present invention described in the claims. Various modifications and changes can be made to the present invention by means of the like, and this will also be said to be included within the scope of the present invention.

Claims (16)

Containing a quinone-based compound represented by the following structural formula 1 as an active ingredient,
The quinone-based compound is a cosmetic composition for skin whitening having a function of reducing melanin accumulation by inhibiting differentiation of melanocytes by inhibiting Wnt/β-catenin signal;
[Structural Formula 1]

In structural formula 1,
n is a repeating number that is an integer selected from 1 to 3,
R ¹ is a C1 to C10 alkyl group,
R ² to R ⁷ are hydrogen atoms. delete The method of claim 1,
The compound represented by the structural formula 1 is a cosmetic composition for skin whitening, characterized in that lucidin-3-methyl ether (lucidin-3-methyl ether). delete delete Containing a quinone-based compound represented by the following structural formula 1 as an active ingredient,
The quinone-based compound is a pharmaceutical composition for preventing or treating hyperpigmentation diseases having a function of reducing melanin accumulation by inhibiting differentiation of melanocytes by inhibiting Wnt/β-catenin signal;
[Structural Formula 1]

In structural formula 1,
n is a repeating number that is an integer selected from 1 to 3,
R ¹ is a C1 to C10 alkyl group,
R ² to R ⁷ are hydrogen atoms. delete The method of claim 6,
The compound represented by the structural formula 1 is a pharmaceutical composition for the prevention or treatment of hyperpigmentation disease, characterized in that lucidin-3-methyl ether (lucidin-3-methyl ether). Containing a quinone-based compound represented by the following structural formula 1 as an active ingredient,
The quinone-based compound is a food composition for preventing or improving hyperpigmentation disease having a function of reducing melanin accumulation by inhibiting differentiation of melanocytes by inhibiting Wnt/β-catenin signal;
[Structural Formula 1]

In structural formula 1,
n is a repeating number that is an integer selected from 1 to 3,
R ¹ is a C1 to C10 alkyl group,
R ² to R ⁷ are hydrogen atoms. delete The method of claim 9,
The compound represented by the structural formula 1 is a food composition for preventing or improving hyperpigmentation disease, characterized in that lucidin-3-methyl ether (lucidin-3-methyl ether). It relates to a method for screening a whitening active compound comprising the step of treating a reporter cell expressing a Wnt/β-catenin signal in a quinone compound library,
The whitening active compound inhibits Wnt/β-catenin signaling,
The screening method uses HEK293-FL (HEK293-Firefly luciferase) reporter cells, which are HEK293 cell lines into which human Frizzled-1 expression plasmid and β-catenin/Tcf-dependent luciferase reporter plasmid are introduced,
The whitening active compound is a method for screening a whitening active compound, characterized in that represented by the following structural formula 1.
[Structural Formula 1]

In structural formula 1,
n is a repeating number that is an integer selected from 1 to 3,
R ¹ is a C1 to C10 alkyl group,
R ² to R ⁷ are hydrogen atoms. The method of claim 12,
The whitening active compound represented by the structural formula 1 is a method for screening a whitening active compound, characterized in that the lucidin-3-methyl ether (lucidin-3-methyl ether). delete delete The method of claim 12,
The HEK293-FL reporter cell is a method for screening a whitening active compound, characterized in that cultured in a Wnt3a-conditioned medium with a compound of a quinone-based compound library.

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