JP6879480B2 - New compounds, cosmetic compositions and pharmaceutical compositions containing them - Google Patents

Description

The present invention relates to novel compounds, cosmetic compositions and pharmaceutical compositions containing them, and more specifically, induces melanosomal degradation by autophagy activation and suppresses endocytosis into keratinocytes. The present invention relates to a novel compound, a cosmetic composition for skin whitening containing the same, and a pharmaceutical composition for preventing and treating an excess-deposited disease of melanin pigment.

Skin color is large and is determined by the amount of melanin, hemoglobin, carotene, etc. Of these, melanin plays the most important role. Melanin not only determines a person's skin color, but also plays a role in protecting the skin, but when it is excessively produced in the skin due to changes in the external environment such as excessive exposure to ultraviolet rays, air pollution, stress, and inflammation, A pigmentation phenomenon may occur, causing darkening of the skin, stains, freckles, etc. Among the above-mentioned external stimulating factors, ultraviolet rays are the largest stimulating source of melanin biosynthesis and may affect various processes related to melanin production. That is, ultraviolet rays act as a main factor for inducing excessive melanin production by promoting the activity of melanocytes, which are melanin-producing cells, promoting the secretion of melanin biosynthesis stimulating hormone, promoting the oxidation of melanin, or promoting the activity of tyrosinase.

Such melanin biosynthesis is carried out by the action of various genes such as tyrosinase, TRP-1 (tyrosinase related protein 1), TRP-2 (tyrosinase related protein 2), and Pmel17 and their corresponding enzymes. In addition, alpha-MSH hormone (α-Melanocyte-stimulating hormone), IL-1, TNF-alpha (tumor necrosis factor-α), GM-CSF (Granulocyte-macrophage color cytokine) mutual cytokine-stimulating Affected by action.

Most whitening materials currently being developed are materials that suppress tyrosinase activity. However, some whitening materials that suppress tyrosinase activity pose safety issues such as vitiligo, irritation and allergic reactions. As an example, hydroxyquinones, which are known to most effectively suppress tyrosinase, are known to have the side effect of killing melanocytes. Therefore, there is an urgent need to develop a whitening material that does not affect the expression of tyrosinase and the melanin content of melanocytes and regulates the endocytosis of melanocytes in keratinocytes.

Autophagy is an energy regeneration and damaging substance that breaks down aging or damaged intracellular substances and organs when the intracellular energy source is depleted or when intracellular stress factors occur excessively. It is a mechanism that eliminates the cells, and means a mechanism that enables the maintenance of normal cells. Recently, through various studies, it has been reported that the intracellular autophagy activity decreases sharply as aging progresses or accelerates aging. That is, when autophagy is suppressed, aged mitochondria and erroneously folded proteins are excessively accumulated in the cell, and free radicals and oxidative stress in the cell increase, which is eventually caused by cell death. It is known that a gene fragment is generated, which stimulates melanocytes, leading to melanosomal maturation and transport to keratinocytes.

That is, autophagy activation decomposes and recycles aging substances and organs in cells, activation of antioxidant proteins prevents cell death, and rapidly removes deformed proteins, lipids, mitochondria, etc. As a result, it provides an environment in which cells can survive in a healthier state and eliminates melanosome maturation factors. Not only that, but recent reports have shown that skin color is regulated by autophagy activation, which brightens the skin color of artificial skin.

In addition, research results show that protease-activated receptor 2 (PAR-2) expressed from keratinocytes is associated with endocytosis influx into keratinocytes of melanosomes, which can regulate pigmentation. Has also been reported. That is, activation of PAR-2 by trypsin induces pigmentation, which can induce hyperpigmentation or uneven skin tone.

Therefore, the present inventors effectively decompose melanosomes in keratinocytes by autophagy activation, effectively suppress skin pigmentation, and promote PAR-2 suppression to promote keratinocyte keratinocytes. The present invention has been completed in order to provide a novel compound capable of effectively suppressing the inflow into the skin and a whitening application containing the same.

Hung et al. Autophagy, 2009, 5, 4, 502-510 Hearing & Jimenez, Int J Biochem, 1987, 19 (12), 1141-7 Funasaka et al. J Lipid Res. 1999, Jul; 40 (7), 1312-6 Qi et al. PLOS one, 2012, 7, 10, e46834 Xilouri et al. Brain, 2013, 136, 2130-2146 Kim et al. The Journal of Clinical Investment, 2014, 124, 8, 3311-3324 Seiberg et al. Experimental Cell Research, 2000, 254, 25-32

An object of the present invention is to provide a novel compound capable of inducing melanosome degradation by autophagy activation or a pharmaceutically acceptable salt thereof.

An object of the present invention is to provide a novel compound or a pharmaceutically acceptable salt thereof capable of reducing the influx of melanosomes by inhibiting PAR-2 activity.

An object of the present invention is to provide a cosmetic composition for skin whitening containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

An object of the present invention is to provide a pharmaceutical composition for ameliorating or treating a hyperdeposited disease of a melanin pigment containing the compound or a pharmaceutically acceptable salt thereof as an active ingredient.

［前記化学式１中、
ＡＡは、置換または非置換のプロリンを含むアミノ酸残基である。］ For the purposes described above, the present invention provides a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof, which can induce melanosomal degradation by autophagy activation.

[In the chemical formula 1,
AA is an amino acid residue containing substituted or unsubstituted proline. ]

In Chemical Formula 1, said AA is an amino acid residue comprising proline substituted or unsubstituted with one or more substituents selected from _{hydroxy, C 1-10} alkoxy, C _{1-10 alkylcarbonyl, carboxy and the like.} May be good.

［前記化学式２中、
Ｒ_１は、Ｃ_１−１０アルキルカルボニルであり；Ｒ_２は、水素、ヒドロキシ、Ｃ_１−１０アルコキシ、Ｃ_１−１０アルキルカルボニルまたはカルボキシである。］ Chemical formula 1 may be a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

[In the chemical formula 2,
R ₁ is a C _1-10 alkyl carbonyl; R ₂ is a hydrogen, hydroxy, C _1-10 alkoxy, C _1-10 alkyl carbonyl or carboxy. ]

［前記化学式３中、
Ｒ_３は、ヒドロキシまたはＣ_１−１０アルキルであり；
Ｒ_４は、水素、ヒドロキシ、Ｃ_１−１０アルコキシ、Ｃ_１−１０アルキルカルボニルまたはカルボキシである。］ Chemical formula 1 may be a compound represented by the following chemical formula 3 or a pharmaceutically acceptable salt thereof.

[In the chemical formula 3,
R ₃ is hydroxy or C _1-10 alkyl;
R ₄ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. ]

For the above-mentioned purposes, the present invention provides a cosmetic composition for skin whitening containing the compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In the skin whitening cosmetic composition, the active ingredient may be selected from the compound represented by the chemical formula 2 or a pharmaceutically acceptable salt (A) thereof.

In the skin whitening cosmetic composition, the active ingredient may be selected from the compound represented by the chemical formula 3 or a pharmaceutically acceptable salt (B) thereof.

In the skin whitening cosmetic composition, the active ingredient is the compound represented by the chemical formula 2 or a pharmaceutically acceptable salt (A) thereof and the compound represented by the chemical formula 3 or a pharmaceutically acceptable salt thereof. It may be a mixture of the salt (B).

In the skin whitening cosmetic composition, the active ingredient may be mixed in a weight ratio of 1: 0.1 to 1: 1 (A: B).

In the skin whitening cosmetic composition, the active ingredient may be contained in an amount of 0.0001 to 10% by weight based on the total weight.

For the above-mentioned purposes, the present invention provides a pharmaceutical composition for improving or treating a hyperdeposited disease of a melanin pigment containing the compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. Will be done.

In a pharmaceutical composition for ameliorating or treating a hyperdeposited melanin pigment disease, the active ingredient may be selected from the compound represented by the chemical formula 2 or a pharmaceutically acceptable salt (A) thereof.

In a pharmaceutical composition for ameliorating or treating a hyperdeposited melanin pigment disease, the active ingredient may be selected from the compound represented by the chemical formula 3 or a pharmaceutically acceptable salt (B) thereof.

In a pharmaceutical composition for improving or treating hyperdeposited melanin pigment, the active ingredient is represented by the compound represented by the chemical formula 2 or a pharmaceutically acceptable salt (A') thereof and the chemical formula 3. The compound may be a mixture thereof or a pharmaceutically acceptable salt (B') thereof.

In a pharmaceutical composition for ameliorating or treating melanin pigment hyperdeposition disease, the active ingredient may be mixed in a weight ratio of 1: 0.1 to 1: 1 (A': B').

In a pharmaceutical composition for ameliorating or treating melanin hyperpigmentation disease, the melanin hyperpigmentation disease is sobacus, senile pigmentation spots, chloasma, spots, brown or black hokuro, sunlight pigmentation spots, blue color. It may be selected from melasma, excessive pigmentation after drug use, melasma gestation and excessive pigmentation after inflammation.

In a pharmaceutical composition for improving or treating an over-deposited disease of melanin pigment, the active ingredient may be contained in an amount of 0.0001 to 10% by weight based on the total weight.

The compounds according to the invention or pharmaceutically acceptable salts thereof can activate autophagy by increasing the expression of autophagy-related proteins and protect cells from oxidative stress. In addition, various diseases and phenomena caused by oxidative stress can be improved, prevented and treated. In particular, autophagy activation, which induces melanosomal degradation and reduces melanin production, has the advantage of achieving a surprising whitening effect.

The compounds according to the invention or pharmaceutically acceptable salts thereof block the influx of melanosomes by inducing melanosome degradation, reducing melanin deposition and effectively suppressing endocytosis into keratinocytes. To do. That is, it is very effective in improving, preventing and treating melanin-induced pigment hyperdeposition disease.

It is the figure which schematized the mechanism which suppresses the pigmentation process of the keratinocyte according to this invention. It is a figure which shows the result of having confirmed whether or not the Example compound by this invention suppresses a tyrosinase activity. It is a figure which shows the result of having performed the MTT assay in the melanocyte (melanocyte) in order to confirm the treatment concentration which does not have cytotoxicity of the Example compound by this invention. It is a figure which shows the result of having performed the MTT assay in the keratinocyte (keratinocyte) in order to confirm the treatment concentration which does not have cytotoxicity of the Example compound by this invention. It is a figure which shows the result of having analyzed whether or not the treatment of the Example compound by this invention affects the expression change of the gene (Tyrosinase) related to melanosome maturation. It is a figure which shows the result of having analyzed whether or not the treatment of the Example compound by this invention affects the expression change of the gene (MITF) associated with melanosome maturation. It is a figure which shows the result of having analyzed the changed melanin content by the treatment of the Example compound by this invention. It is a figure which shows the result of having analyzed the expression change of the protein related to autophagy activation, and the change of mRNA of a gene related to endocytosis by the treatment of the Example compound by the present invention. It is a figure which shows the result of the comparative analysis of the influence on the melanin deposition by the treatment of the Example compound by this invention. By the treatment of the example compound according to the present invention, it was confirmed whether or not the melanin deposition inhibitory ability was dependent on autophagy, and the melanin content and autophagy activity changed in the keratinized cells in which ATG5 was knock-downed. It is a figure which shows the result of comparative analysis of the change of a melanin-related protein. It is a figure which shows the result of having analyzed the melanosomes decomposition by the activated autophagy by the treatment of the Example compound by this invention with a confocal microscope. It is a figure which shows the result of having quantitatively analyzed the melanosomes decomposition by the activated autophagy by the treatment of the Example compound by this invention with an electron microscope (scale bar: 2 μm / 500 nm / 2 μm / 500 nm). It is a figure which shows the result of having quantitatively analyzed the deposition suppression and decomposition of melanosomes by the treatment of the Example compound by this invention. It is a figure which shows the result of having confirmed whether the treatment of the Example compound by this invention reduces melanin deposition in an artificial skin model. It is a figure which shows the result of having quantitatively analyzed the reduction of the melanin deposition in the artificial skin model by the treatment of the Example compound by this invention by F & M staining.

The novel compounds according to the invention, cosmetic compositions and pharmaceutical compositions containing them will be described in detail below, but unless otherwise defined in the technical and scientific terms used herein, the techniques to which the invention belongs. It has a meaning that is usually understood by a person having ordinary knowledge in the field, and the description of a known function and configuration that may obscure the gist of the present invention is omitted in the following description.

As used herein, substituents containing "alkyl," "alkoxy," and other "alkyl" moieties include either straight-chain or branched-chain forms. Further, as the alkyl, alkoxy and the like according to the present invention, those having a linear carbon number of 1 to 10 or those having a linear carbon number are given priority, but those having a carbon number of 11 to 30 are also present. Needless to say, this is one aspect of the invention.

As used herein, "alkylcarbonyl" means * -C (= O) alkyl, where the alkyl follows the above definition.

As used herein, a "pharmaceutically acceptable salt" is produced using conventional techniques known in the art and is a pharmaceutically acceptable inorganic acid, organic acid or Contains salts derived from bases. Examples of the inorganic or organic acid include hydrochloric acid, bromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, and the like. Examples thereof include tartrate acid, acetic acid, trifluoroacetic acid, citric acid, methanesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid and benzenesulfonic acid. In addition, examples of the base include alkali metals such as potassium and sodium; alkaline earth metals such as magnesium; or ammonium; and the like.

As used herein, "improvement" refers to any act of alleviating or improving or favorably changing the condition by applying the composition according to the invention.

In addition, the singular form used herein may be intended to include a plurality of forms unless otherwise specified in the context.

In addition, the unit used in this specification without particular limitation is based on weight, and as an example, the unit of% or ratio means weight% or weight ratio.

Also, the numerical ranges used herein are double limited, with lower and upper limits and all values within that range, increments logically derived from the form and width of the defined range. Includes all possible combinations of upper and lower limits for all values and numerical ranges limited to different forms.

Further, in the present specification, the expression "includes" is an open type description having the same meaning as the expressions such as "provide", "include", "have", or "feature", and further mentions them. Do not exclude elements, materials or processes that are not.

After extensive research on whitening materials, the present inventors have devised a novel compound capable of effectively inducing melanosomal degradation by autophagy activation. The compound according to the present invention has an excellent autophagy activation ability, and is expected to be actively utilized for amelioration, prevention and treatment of diseases caused by excessive deposition of melanin pigment.

Specifically, the compounds according to the invention increase the expression of autophagy activation-related proteins, thereby inducing melanosome degradation. Thereby, the production of melanosomes can be effectively suppressed. Therefore, a composition containing this as an active ingredient can exhibit excellent effects in amelioration, prevention and treatment of melanin pigment hyperdeposition disease, not to mention a whitening effect.

At the same time, the compound according to the present invention suppresses the activation of protease-activated receptor 2 (PAR-2) expressed in keratinocytes, and effectively suppresses endocytosis of keratinocytes. Therefore, the influx of melanosomes can be effectively blocked.

Hereinafter, the present invention will be specifically described.

［前記化学式１中、
ＡＡは、置換または非置換のプロリンを含むアミノ酸残基である。］ The compound according to one embodiment of the present invention may be represented by the following chemical formula 1 or may be a pharmaceutically acceptable salt thereof.

[In the chemical formula 1,
AA is an amino acid residue containing substituted or unsubstituted proline. ]

The compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof activates intracellular autophagy, induces melanosomal degradation, and effectively suppresses the expression of MITF (Melanogenesis Associated Transcription Factor). To do. In addition, it suppresses melanosomal maturation of melanocytes and suppresses endocytosis in keratinocytes, thereby realizing a surprising whitening effect. That is, the compound represented by the chemical formula 1 according to the present invention can be usefully utilized as a whitening material having an autophagy activation function.

Intracellular autophagy activation occurs vigorously in young human tissues and cells, but as aging progresses, the expression level of intracellular autophagy-related proteins decreases sharply, resulting in a sharp decrease in autophagy activity. To do. Therefore, intracellular aging proteins, lipids and mitochondria are not removed in a timely manner, and the phenomenon of cellular senescence occurs rapidly. According to the present invention, autophagy activation suppresses aging of each cell, each tissue and each individual from the beginning, and enables prevention, amelioration and treatment of various diseases caused by aging.

In addition, activation of autophagy improves the viability of aged cells by removing harmful proteins and organs in the cells, and is also very closely related to the increase in lifespan of each individual.

Therefore, the compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof suppresses the maturation of melanosomes in melanocytes due to various causes, and is used for the prevention and treatment of melanin pigment hyperdeposition disease. It works more usefully.

In Chemical Formula 1, the AA is an amino acid residue comprising proline substituted or unsubstituted with one or more substituents selected from _{hydroxy, C 1-10} alkoxy, C _{1-10 alkyl carbonyl and carboxy.} May be good.

As an example, in the chemical formula 1, the AA may be an amino acid residue containing proline linked by an amide bond. At this time, the proline and amino acid residues other than proline may be substituted or unsubstituted with the above-mentioned substituents. Further, the amino acid residue may contain proline and two or more amino acids may be bound by an amide bond.

As an example, in the chemical formula 1, the AA is an amino acid residue containing proline linked by an amide bond, and the amino acid residue is selected from Asn, Gln, Lys and the like having two N-terminals. It may be an amino acid residue containing one or more of them.

As an example, in the above chemical formula 1, the AA may be an amino acid residue to which proline substituted or unsubstituted by the above-mentioned substituent is directly bound.

［前記化学式２中、
Ｒ_１は、Ｃ_１−１０アルキルカルボニルであり；
Ｒ_２は、水素、ヒドロキシ、Ｃ_１−１０アルコキシ、Ｃ_１−１０アルキルカルボニルまたはカルボキシである。］ The compound according to one embodiment of the present invention may be a compound represented by the following chemical formula 2.

[In the chemical formula 2,
R ₁ is a C _1-10 alkyl carbonyl;
R ₂ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. ]

The compound represented by the chemical formula 2 is excellent in the melanosome degradation effect by autophagy activation, and can mean a melanososome degradation-inducing compound by autophagy activation in the present specification.

As an example, in said Chemical Formula 2, said R ₁ is a C _1-4 alkylcarbonyl; said R ₂ may be hydrogen, hydroxy or C _1-4 alkoxy.

As an example, in said Chemical Formula 2, said R ₁ is a linear C _1-4 alkylcarbonyl; said R ₂ may be hydrogen or hydroxy.

As an example, in the chemical formula 2, the R ₁ is a C _1-2 alkylcarbonyl; the R ₂ may be hydrogen or hydroxy.

Further, the compound represented by the chemical formula 2 contains one or more chiral asymmetric carbon atoms. Thereby, the compound represented by the chemical formula 2 can exist in a racemic form and an optically active form. That is, the diastereomers and enantiomers of the compounds represented by the chemical formula 2 are both included in the category of the present invention.

［前記化学式３中、
Ｒ_３は、ヒドロキシまたはＣ_１−１０アルキルであり；
Ｒ_４は、水素、ヒドロキシ、Ｃ_１−１０アルコキシ、Ｃ_１−１０アルキルカルボニルまたはカルボキシである。］ The compound according to one embodiment of the present invention may be a compound represented by the following chemical formula 3.

[In the chemical formula 3,
R ₃ is hydroxy or C _1-10 alkyl;
R ₄ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. ]

The compound represented by the chemical formula 3 can effectively suppress the influx of melanosomes in keratinocytes by suppressing endocytosis, and in the present specification, it means a compound that induces the suppression of melanosomes by suppressing endocytosis. obtain.

As an example, in said Chemical Formula 3, said R ₃ may be hydroxy or C _1-4 alkyl; said R ₄ may be hydrogen, hydroxy or C _1-4 alkoxy.

As an example, in said Chemical Formula 3, said R ₃ may be hydroxy or linear C _1-4 alkyl; said R ₄ may be hydrogen, hydroxy or straight chain C _1-4 alkoxy.

As an example, in said Chemical Formula 3, said R ₃ may be hydroxy or C _1-2 alkyl; said R ₄ may be hydrogen or hydroxy.

Further, the compound represented by the chemical formula 3 contains one or more chiral asymmetric carbon atoms. Thereby, the compound represented by the chemical formula 3 can exist in a racemic form and an optically active form. That is, the diastereomers and enantiomers of the compounds represented by the chemical formula 3 are both included in the category of the present invention.

Hereinafter, the uses of the present invention will be specifically described.

One aspect of the application according to one embodiment of the present invention may be a skin whitening cosmetic composition.

The skin whitening cosmetic composition according to one embodiment of the present invention may contain the compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof does not induce cytotoxicity within the concentration range showing activity, and exhibits a whitening effect even at a very low concentration range (for example, 1 μm). Further, the compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof does not reduce the stability and storage stability of the dosage form even when it is formulated into a normal cosmetic composition.

The skin whitening cosmetic composition according to one embodiment of the present invention may contain, as an active ingredient, the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt (A) thereof. At this time, the above (A) may be one or a mixture of two or more.

The skin whitening cosmetic composition according to one embodiment of the present invention may contain, as an active ingredient, the compound represented by the above chemical formula 3 or a pharmaceutically acceptable salt (B) thereof. At this time, the above (B) may be one or a mixture of two or more.

The skin whitening cosmetic composition according to one embodiment of the present invention contains the compound represented by the chemical formula 2 or a pharmaceutically acceptable salt (A) thereof and the compound represented by the chemical formula 3 as an active ingredient. It may be a mixture containing the pharmaceutically acceptable salt (B).

The skin whitening cosmetic composition according to one embodiment of the present invention contains 0.0001 to 10% by weight of the active ingredient with respect to the total weight in terms of realizing a substantial autophagy activating effect. It may be. Considering the effect on the content, the active ingredient is specifically 0.0005 to 5% by weight, more specifically 0.001 to 1% by weight, and most specifically 0.001 to 0.1% by weight. May be included.

Furthermore, in order to realize a more improved whitening effect, the cosmetic composition for skin whitening according to one embodiment of the present invention contains a melanosomes decomposition-inducing compound by autophagy activation and a melanosomes inflow suppression-inducing compound by endocytosis suppression. The mixture can be simultaneously included as an active ingredient.

As an example, the melanosomes degradation-inducing compound by autophagy activation may be at least one compound (A) selected from the compounds represented by the chemical formula 2, and the melanosome influx inhibition induction by the endocytosis inhibition may be used. The compound may be at least one compound (B) selected from the compounds represented by the chemical formula 3.

The skin whitening cosmetic composition according to one embodiment of the present invention comprises the melanosome decomposition-inducing compound by autophagy activation and suppression of melanosome influx by suppressing endocytosis at an appropriate weight ratio according to the application form, purpose of use and desired effect. It goes without saying that a mixture of inducible compounds can be used.

As an example, the mixture may be mixed in a weight ratio of 1: 0.1 to 1: 1 (A: B).

As an example, the mixture may be mixed in a weight ratio (A: B) of 1: 0.1 to 1: 0.8.

As an example, the mixture may be mixed in a weight ratio of 1: 0.1 to 1: 0.5 (A: B).

The skin whitening cosmetic composition can be formulated into a general emulsifier form, a solubilizer form, or the like by using a generally known production method.

As an example, the skin whitening cosmetic composition is an agent selected from the group composed of softening lotion, astringent lotion, nutritional lotion, eye cream, nutritional cream, massage cream, powder, essence, facial mask and the like. It may be dosaged into a form.

As an example, the skin whitening cosmetic composition may be formulated into a dosage form selected from the group composed of soap, cleansing cream, cleansing foam, cleansing water and the like.

In addition, the skin whitening cosmetic composition may further contain additional additives as appropriate, depending on the purpose. As an example, the additives include stabilizers, emulsifiers, thickeners, moisturizers, liquid crystal film strengtheners, pH regulators, antibacterial agents, water-soluble polymers, coating agents, metal ion sequestering agents, amino acids, organic amines. One or more aqueous additives selected from, polymer emulsions, pH regulators, skin nutrients, antioxidants, antioxidant aids, preservatives, fragrances, etc .; fats and oils, waxes, hydrocarbon oils, One or more oily additives selected from higher fatty acid oils, higher alcohols, synthetic ester oils, silicone oils and the like; and the like.

At this time, each of the additives may be contained in an amount of 0.001 to 20% by weight, specifically 0.01 to 10% by weight and 0.05 to 10% by weight, based on the total weight of the composition. It may be included, but is not limited to this.

Another use of one embodiment of the present invention may be a pharmaceutical composition for ameliorating or treating hyperdeposited melanin pigments.

In addition, one aspect of the use according to one embodiment of the present invention may be a method for alleviating or treating an overdeposited disease of melanin pigment. Specifically, the method may be to apply an effective amount of the compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof to the skin of a subject in need thereof, which is required. The skin of the subject may be the skin of the melanin pigment hyperdeposition disease.

At this time, the effective amount is not limited as long as it is within the range of the amount of the active ingredient used.

The pharmaceutical composition for improving or treating the hyperdeposited disease of melanin pigment according to one embodiment of the present invention may contain the compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof exhibits an excellent effect in ameliorating, preventing and treating an over-deposited disease of melanin pigment by realizing an excellent whitening effect. Specifically, the compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof can increase the expression of autophagy-related proteins, activate autophagy, and protect cells from oxidative stress. , Various diseases and phenomena caused by oxidative stress can be improved, prevented and treated. In particular, it induces melanosome degradation to reduce melanin production, suppresses melanosome maturation, effectively suppresses endocytosis of keratinocytes, blocks melanosome influx, and is a pigment derived from melanin. It is very effective in improving, preventing and treating hyperdeposited diseases.

The pigmented hyperpigmentation diseases are freckles; senile pigmented spots; chloasma; spots; brown or black spots; sun-induced pigmented spots; blue melanosis; excessive pigmentation after drug use; gestational brown spots; Alternatively, it may include, but is not limited to, excessive pigmentation after inflammation;

The pharmaceutical composition for improving or treating melanin pigment hyperdeposition disease according to one embodiment of the present invention contains, as an active ingredient, the compound represented by the above chemical formula 2 or a pharmaceutically acceptable salt (A') thereof. It may be. At this time, the (A') may be one or a mixture of two or more.

The pharmaceutical composition for improving or treating melanin pigment hyperdeposition disease according to one embodiment of the present invention contains, as an active ingredient, the compound represented by the above chemical formula 3 or a pharmaceutically acceptable salt (B') thereof. It may be. At this time, the (B') may be one or a mixture of two or more.

The pharmaceutical composition for improving or treating hyperdeposited melanin pigment according to one embodiment of the present invention contains the compound represented by the chemical formula 2 or a pharmaceutically acceptable salt (A') thereof as an active ingredient. It may be a compound containing the compound represented by Chemical Formula 3 or a pharmaceutically acceptable salt (B') thereof.

The pharmaceutical composition for improving or treating the hyperdeposited disease of melanin pigment according to one embodiment of the present invention contains the active ingredient based on the total weight in terms of achieving a substantial autophagy activating effect. It may contain 0.0001 to 10% by weight. Considering the effect on the content, the active ingredient is specifically 0.001 to 5% by weight, more specifically 0.001 to 1% by weight, and most specifically 0.001 to 0.1% by weight. May be included.

Furthermore, the pharmaceutical composition for improving or treating the hyperdeposited disease of melanin pigment according to one embodiment of the present invention suppresses melanosome degradation-inducing compounds and endocytosis by autophagy activation in order to realize a further improved effect. A mixture of melanosome inflow suppression-inducing compounds according to the above can be contained as an active ingredient.

As an example, the melanosomes degradation-inducing compound by autophagy activation may be at least one compound (A') selected from the compounds represented by the chemical formula 2, and the melanosome influx is suppressed by suppressing endocytosis. The inducing compound may be at least one compound (B') selected from the compounds represented by the chemical formula 3.

The pharmaceutical composition for improving or treating melanin pigment hyperdeposition disease according to one embodiment of the present invention induces melanosome degradation by autophagy activation in an appropriate weight ratio according to an application form, intended use and desired effect. It goes without saying that a mixture of the compound and the compound that induces the suppression of melanosome influx by suppressing endocytosis can be used.

As an example, the mixture may be mixed in a weight ratio of 1: 0.1 to 1: 1 (A': B').

As an example, the mixture may be mixed in a weight ratio of 1: 0.1 to 1: 0.8 (A': B').

As an example, the mixture may be mixed in a weight ratio of 1: 0.1 to 1: 0.5 (A': B').

The pharmaceutical composition for ameliorating or treating a hyperdeposited melanin pigment disease can be formulated into a dosage form containing a pharmaceutically acceptable carrier using a commonly known production method. ..

As an example, the pharmaceutical composition for ameliorating or treating an overdeposited melanin pigment disease is formulated into a topical skin formulation selected from the group consisting of lotions, ointments, gels, creams, patches, sprays and the like. It may be converted.

As an example, the pharmaceutical compositions for improving or treating hyperdeposited melanin pigments are oral (tablets, capsules, powders), oral, sublingual, rectal, vaginal, intranasal, topical or non-existent. It may be formulated into an oral (including intravenous, cavernous, intramuscular, subcutaneous and intraductal) dosage form.

In addition, the pharmaceutical composition for improving or treating the hyperdeposited disease of the melanin pigment may appropriately contain an additional pharmaceutically acceptable carrier depending on the purpose. As an example, lactose, dextros, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxy It may be, but is not limited to, benzoate, talc, magnesium stearate or mineral oil. In addition to the above carriers, carriers such as lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents or preservatives may be further contained.

At this time, each of the carriers may be contained in an amount of 0.001 to 20% by weight, specifically 0.01 to 10% by weight and 0.05 to 10% by weight, based on the total weight of the composition. However, it is not limited to this.

Hereinafter, the present invention will be described in detail with reference to Examples. However, the following examples merely exemplify the present invention, and the present invention is not limited to the following examples.

１０ｍｌの反応容器に２−クロロトリチルクロライド樹脂（２−ｃｈｌｏｒｏ ｔｒｉｔｙｌ ｃｈｌｏｒｉｄｅ ｒｅｓｉｎ、１００−２００ｍｅｓｈ、Ｎｏｖａｂｉｏｃｈｅｍ１００ｍｇ、１当量）とＦｍｏｃ−Ｌｙｓ（Ｄｄｅ）−ＯＨ（Ｎα−Ｆｍｏｃ−Ｎε−Ｄｄｅ−Ｌ−リジン、Ｎα−Ｆｍｏｃ−Ｎε−［１−（４，４−ジメチル−２，６−ジオキソシクロヘキシリデン）エチル］−Ｌ−リジン）（１０６．５ｍｇ、２当量）およびジイソプロピルアミン（ＤＩＰＥＡ、６９．７μｌ、４当量）を５ｍｌのジクロロメタン（ＤＣＭ）に溶解して添加し、常温（２３℃）で１２時間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのジクロロメタンとメタノール（ＭｅＯＨ）、ジクロロメタンおよびジメチルホルムアミド（ＤＭＦ）を用いて順に洗浄し、化合物１ａを定量的に得た。 (Example 1) Synthesis of compound 1 Step 1. Synthesis of compound 1a (Fmoc-Lys (Dde) -O-2-chlorotrityl resin)

2-Chlorotrityl chloride resin, 100-200 mesh, Novabiochem 100 mg, 1 equivalent) and Fmoc-Lys (Dde) -OH (Nα-Fmoc-Nε-Dde-L-lysine, in a 10 ml reaction vessel. Nα-Fmoc-Nε- [1- (4,4-dimethyl-2,6-dioxocyclohexylidene) ethyl] -L-lysine) (106.5 mg, 2 equivalents) and diisopropylamine (DIPEA, 69.7 μl) 4 equivalents) was dissolved in 5 ml of dichloromethane (DCM), added, and reacted at room temperature (23 ° C.) for 12 hours. The reaction mixture was removed by filtration, and the synthesized resin was washed successively with 10 ml of dichloromethane and methanol (MeOH), dichloromethane and dimethylformamide (DMF), respectively, to obtain compound 1a quantitatively.

ステップ１の反応容器（ステップ１で得られた化合物１ａ）に５ｍｌの２０％ピペリジン（ｉｎ ＤＭＦ）を入れて常温で５分間反応させた後、濾過して反応液を除去した。５ｍｌの２０％ピペリジン（ｉｎ ＤＭＦ）をさらに加え、常温で５分間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄した後、Ｆｍｏｃ−Ｌｙｓ（Ｆｍｏｃ）−ＯＨ（２３６．３ｍｇ、４当量）とＨＯＢｔ（ヒドロキシベンゾトリアゾール、５４．１ｍｇ、４当量）およびＮ，Ｎ−ジイソプロピルカルボジイミド（ＤＩＣ）（６１．９μｌ、４当量）を５ｍｌのＤＭＦに溶解して加え、常温で２時間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄し、化合物１ｂを定量的に得た。 Step 2. Synthesis of compound 1b (Fmoc-Lys (Fmoc) -Lys (Dde) -O-2-chlorotrityl resin)

5 ml of 20% piperidine (in DMF) was placed in the reaction vessel of step 1 (compound 1a obtained in step 1), reacted at room temperature for 5 minutes, and then filtered to remove the reaction solution. 5 ml of 20% piperidine (in DMF) was further added and reacted at room temperature for 5 minutes. The reaction solution was removed by filtration, and the synthesized resin was washed successively with 10 ml of DCM and MeOH, DCM and DMF, respectively, followed by Fmoc-Lys (Fmoc) -OH (236.3 mg, 4 eq) and HOBt. (Hydroxybenzotriazole, 54.1 mg, 4 eq) and N, N-diisopropylcarbodiimide (DIC) (61.9 μl, 4 eq) were added dissolved in 5 ml of DMF and reacted at room temperature for 2 hours. The reaction mixture was removed by filtration, and the synthesized resin was washed successively with 10 ml of DCM and MeOH, DCM and DMF, respectively, to obtain compound 1b quantitatively.

ステップ２の反応容器（ステップ２で得られた化合物１ｂ）に５ｍｌの２０％ピペリジン（ｉｎ ＤＭＦ）を入れて常温で５分間反応させた後、濾過して反応液を除去した。５ｍｌの２０％ピペリジン（ｉｎ ＤＭＦ）をさらに加え、常温で５分間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ５００ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄した後、ｔ−ブチルブロモアセテート（ｔｅｒｔ−ｂｕｔｙｌ ｂｒｏｍｏａｃｅｔａｔｅ、１４７．７μｌ、１０当量）とＤＩＰＥＡ（２６１．３μｌ、１５当量）を５ｍｌのＤＭＦに溶解して加え、常温で４８時間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄し、化合物１ｃを定量的に得た。 Step 3. Synthesis of compound 1c (tert-butoxycarbonylmethyl) 2-Lys (tert-butoxycarbonylmethyl) 2-Lys (Dde) -O-2-chlorotrityl resin)

5 ml of 20% piperidine (in DMF) was placed in the reaction vessel of step 2 (compound 1b obtained in step 2), reacted at room temperature for 5 minutes, and then filtered to remove the reaction solution. 5 ml of 20% piperidine (in DMF) was further added and reacted at room temperature for 5 minutes. The reaction mixture was removed by filtration, and the synthesized resin was washed successively with 500 ml of DCM and MeOH, DCM and DMF, respectively, and then t-butyl bromoacetate (147.7 μl, 10 eq). And DIPEA (261.3 μl, 15 eq) were dissolved in 5 ml of DMF, added, and reacted at room temperature for 48 hours. The reaction mixture was removed by filtration, and the synthesized resin was washed successively with 10 ml of DCM and MeOH, DCM and DMF, respectively, to obtain compound 1c quantitatively.

ステップ３の反応容器（ステップ３で得られた化合物１ｃ）に５ｍｌの２％ヒドラジン（ｉｎ ＤＭＦ）を入れて５分間常温で反応させた。濾過して反応液を除去し、合成された樹脂を１０ｍｌ ＤＭＦを用いて洗浄した後、１０ｍｌの１０％ＤＩＰＥＡ（ｉｎ ＤＭＦ）を加え、常温で５分間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄し、化合物１ｄを定量的に得た。 Step 4. Synthesis of compound 1d (tert-butoxycarbonylmethyl) 2-Lys (tert-butoxycarbonylmethyl) 2-Lys (NH2) -O-2-chlorotrityl resin)

5 ml of 2% hydrazine (in DMF) was placed in the reaction vessel of step 3 (compound 1c obtained in step 3) and reacted at room temperature for 5 minutes. The reaction solution was removed by filtration, and the synthesized resin was washed with 10 ml DMF, 10 ml of 10% DIPEA (in DMF) was added, and the mixture was reacted at room temperature for 5 minutes. The reaction mixture was removed by filtration, and the synthesized resin was washed successively with 10 ml of DCM and MeOH, DCM and DMF, respectively, to obtain compound 1d quantitatively.

ステップ４の反応容器（ステップ４で得られた化合物１ｄ）にＮ−アセチル−Ｌ−ヒドロキシプロリン（Ｎ−Ａｃｅｔｙｌ−Ｌ−ｈｙｄｒｏｘｙｐｒｏｌｉｎｅ、１３８．５ｍｇ、８当量）とＨＯＢｔ（１８０．１ｍｇ、８当量）およびＤＩＣ（１２３．９μｌ、８当量）を５ｍｌのＤＭＦに溶解して加え、常温で３時間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＤＭＦ、ＤＣＭを用いて順に洗浄した。 Step 5. Synthesis of compound 1

N-Acetyl-L-hydroxyproline (N-Acetyl-L-hydroxyproline, 138.5 mg, 8 eq) and HOBt (180.1 mg, 8 eq) in the reaction vessel of step 4 (Compound 1d obtained in step 4). And DIC (123.9 μl, 8 eq) were dissolved in 5 ml of DMF and added and reacted at room temperature for 3 hours. The reaction solution was removed by filtration, and the synthesized resin was washed sequentially with 10 ml of DCM, DMF, and DCM, respectively.

After vacuum drying, 5 ml of a clear cocktail (trifluoroacetic acid: triisopropylsilane: DW = 95: 2.5: 2.5) was added, and the mixture was reacted at room temperature for 3 hours. The reaction was collected by filtration and 45 ml of diethyl ether was added thereto to precipitate the product. The solid product was collected using a centrifuge and washed twice with 45 ml diethyl ether. The obtained solid product was purified by Prep-HPLC (volume C18, 10 μm, 250 mm × 22 mm) and then lyophilized to yield 55% of compound 1 (molecular weight measured by LC mass: 661.66). A rate of 36.4 mg was obtained.

１０ｍｌの反応容器に２−クロロトリチルクロライド樹脂（１００−２００ｍｅｓｈ、Ｎｏｖａｂｉｏｃｈｅｍ ２００ｍｇ、１当量）とＦｍｏｃ−Ｈｙｐ（ｔＢｕ）−ＯＨ（Ｆｍｏｃ−Ｏ−ｔｅｒｔ−ブチル−Ｌ−ヒドロキシプロリン）（１６３．８ｍｇ、２当量）およびＤＩＰＥＡ（１３９．３μｌ、４当量）を５ｍｌのＤＣＭに溶解して添加し、常温で１２時間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄し、化合物２ａを定量的に得た。 (Example 2) Synthesis of compound 2 Step 1. Synthesis of compound 2a (Fmoc-Hyp (tBu) -O-2-chlorotrityl resin)

2-Chlorotrityl chloride resin (100-200 mesh, Novabiochem 200 mg, 1 eq) and Fmoc-Hyp (tBu) -OH (Fmoc-O-tert-butyl-L-hydroxyproline) (163.8 mg, in a 10 ml reaction vessel. 2 equivalents) and DIPEA (139.3 μl, 4 equivalents) were dissolved in 5 ml DCM, added and reacted at room temperature for 12 hours. The reaction solution was removed by filtration, and the synthesized resin was washed successively with 10 ml of DCM and MeOH, DCM and DMF, respectively, to obtain compound 2a quantitatively.

ステップ１の反応容器（ステップ１で得られた化合物２ａ）に５ｍｌの２０％ピペリジン（ｉｎ ＤＭＦ）を入れて常温で５分間反応させた後、濾過して反応液を除去した。５ｍｌの２０％ピペリジン（ｉｎ ＤＭＦ）をさらに加え、常温で５分間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄した後、Ｆｍｏｃ−Ｌｙｓ（Ｆｍｏｃ）−ＯＨ（４７２．６ｍｇ、４当量）とＨＯＢｔ（１０８．１ｍｇ、４当量）およびＤＩＣ（１２３．９μｌ、４当量）を５ｍｌのＤＭＦに溶解して加え、常温で２時間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄し、化合物２ｂを定量的に得た。 Step 2. Synthesis of compound 2b (Fmoc-Lys (Fmoc) -Hyp (tBu) -O-2-chlorotrityl resin)

5 ml of 20% piperidine (in DMF) was placed in the reaction vessel of step 1 (compound 2a obtained in step 1), reacted at room temperature for 5 minutes, and then filtered to remove the reaction solution. 5 ml of 20% piperidine (in DMF) was further added and reacted at room temperature for 5 minutes. The reaction solution was removed by filtration, and the synthesized resin was washed sequentially with 10 ml of DCM and MeOH, DCM and DMF, respectively, and then Fmoc-Lys (Fmoc) -OH (472.6 mg, 4 eq) and HOBt. (108.1 mg, 4 eq) and DIC (123.9 μl, 4 eq) were dissolved in 5 ml of DMF, added, and reacted at room temperature for 2 hours. The reaction mixture was removed by filtration, and the synthesized resin was washed successively with 10 ml of DCM and MeOH, DCM and DMF, respectively, to obtain compound 2b quantitatively.

ステップ２の反応容器（ステップ２で得られた化合物２ｂ）に５ｍｌの２０％ピペリジン（ｉｎ ＤＭＦ）を入れて常温で５分間反応させた後、濾過して反応液を除去した。５ｍｌの２０％ピペリジン（ｉｎ ＤＭＦ）をさらに加え、常温で５分間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ５００ｍｌのＤＣＭとＭｅＯＨ、ＤＣＭおよびＤＭＦを用いて順に洗浄した後、ｔ−ブチルブロモアセテート（２９５．３μｌ、１０当量）とＤＩＰＥＡ（５２２．５μｌ、１５当量）を５ｍｌのＤＭＦに溶解して加え、常温で４８時間反応させた。濾過して反応液を除去し、合成された樹脂をそれぞれ１０ｍｌのＤＣＭとＤＭＦおよびＤＣＭを用いて順に洗浄し、化合物２ｃを定量的に得た。 Step 3. Synthesis of compound 2c (tert-butoxycarbonylmethyl) 2-Lys (tert-butoxycarbonylmethyl) 2-Hyp (tBu) -O-2-chlorotrityl resin)

5 ml of 20% piperidine (in DMF) was placed in the reaction vessel of step 2 (compound 2b obtained in step 2), reacted at room temperature for 5 minutes, and then filtered to remove the reaction solution. 5 ml of 20% piperidine (in DMF) was further added and reacted at room temperature for 5 minutes. The reaction mixture was removed by filtration, and the synthesized resin was washed successively with 500 ml of DCM and MeOH, DCM and DMF, respectively, followed by t-butylbromoacetate (295.3 μl, 10 eq) and DIPEA (522. (5 μl, 15 eq) was dissolved in 5 ml of DMF, added, and reacted at room temperature for 48 hours. The reaction mixture was removed by filtration, and the synthesized resin was washed successively with 10 ml of DCM, DMF and DCM, respectively, to obtain compound 2c quantitatively.

ステップ３の反応容器（ステップ３で得られた化合物２ｃ）に５ｍｌのｃｌｅａｖａｇｅ ｃｏｃｋｔａｉｌ（トリフルオロ酢酸：トリイソプロピルシラン：ＤＷ＝９５：２．５：２．５）を加え、常温で３時間反応させた。濾過して反応液を集め、これに４５ｍｌのジエチルエーテルを加えて生成物を沈殿させた。遠心分離機を用いて固体生成物を集め、４５ｍｌのジエチルエーテルで２回洗浄した。得られた固体生成物をＰｒｅｐ−ＨＰＬＣ（ｃｏｌｕｍｎ Ｃ１８、１０μｍ、２５０ｍｍ×２２ｍｍ）用いて精製した後、凍結乾燥して化合物２（ＬＣ ｍａｓｓで測定された分子量：４９１．４５）を６０％全収率で５９ｍｇ得た。 Step 4. Synthesis of compound 2

5 ml of clearavage cocktail (trifluoroacetic acid: triisopropylsilane: DW = 95: 2.5: 2.5) was added to the reaction vessel of step 3 (compound 2c obtained in step 3), and the mixture was reacted at room temperature for 3 hours. It was. The reaction was collected by filtration and 45 ml of diethyl ether was added thereto to precipitate the product. The solid product was collected using a centrifuge and washed twice with 45 ml diethyl ether. The obtained solid product was purified by Prep-HPLC (volume C18, 10 μm, 250 mm × 22 mm) and then lyophilized to yield 60% of compound 2 (molecular weight measured by LC mass: 491.45). A rate of 59 mg was obtained.

(Example 3) Confirmation of tyrosinase inhibitory ability In order to analyze whether compound 1 and compound 2 obtained in the above example suppress tyrosinase activity, a tyrosinase inhibition assay was performed.

As a specific experimental method, mushroom tyrosinase was diluted with 2 unit / μl in 0.1 M sodium phosphate buffer and dissolved in water at 0.03% by weight of tyrosine. .. Each experimental group shown in Table 1 below was also diluted with 0.1 M sodium phosphate buffer at 10 ppm, 100 ppm, 1000 ppm, and 10000 ppm, and mixed as shown in Table 1. The reaction was carried out at 37 ° C. for 1 hour, and the absorbance was measured (wavelength: 490 nm, Epoch, manufactured by Biotech). Based on the results, the tyrosinase inhibition rate for the control was calculated.

The result is shown in FIG.

As shown in FIG. 2, it was confirmed that Compound 1 and Compound 2 did not suppress tyrosinase activity at all concentrations.

(Example 4) Confirmation of appropriate treatment concentration In order to confirm the treatment concentration of compound 1 and compound 2 obtained in the above example without cytotoxicity, an MTT assay was performed.

As a specific experimental method, in order to confirm the cell toxicity depending on the treatment concentration of Compound 1 and Compound 2, melanocytes (human primary melanocyte, HEMn-DP, cat # C2025C, manufactured by Gibco) and keratinized cells (human primary). Cell death analysis was performed with keratinocyte, HEKn, cat # C0015C, manufactured by Gibco). The number of cells was dispensed into a 96-well plate at 1 × 10 ⁴ cells / well, and cultured in an incubator for 24 hours at 37 ° C. and 5% CO _2.

Each of Compound 1 and Compound 2 obtained in the above example was dissolved in distilled water (DW) at a concentration of 100 mM to prepare a concentrated solution, which was diluted with a medium and treated at concentrations of 5 μM, 10 μM and 50 μM. After 4 days, the medium on the plate was removed and treated with 100 μl of MTT reagent (Sigma-Aldrich, 0.5 mg / ml). After 4 hours, the reagent was removed, dimethyl sulfoxide (DMSO) was added, and then the absorbance was measured (wavelength: 570 nm, Epoch, manufactured by Biotech). At this time, the cell viability was calculated as a percentage of the absorbance of the control (Blank) not treated with the compound.

The results are shown in FIGS. 3 and 4.

As shown in FIGS. 3 and 4, it was confirmed that Compound 1 and Compound 2 had almost no cytotoxicity even at concentrations of 5 μM, 10 μM, and 50 μM. That is, it was confirmed that the compound according to the present invention is not cytotoxic in either melanocytes or keratinocytes at the above concentration. Therefore, in the subsequent experiments, the experiments were performed at a concentration of 10 μM.

(Example 5) Confirmation of changes in melanosome maturation-related genes and changes in melanin content In order to analyze how Compound 1 and Compound 2 obtained in the above Examples affect melanosome maturation. After treating Compound 1 and Compound 2, the amount of melanosome maturation-related mRNA was analyzed by QPCR, and the melanin content was measured by absorbance.

As a specific experimental method, human-derived melanocytes (HEMn-DP, cat # C2025C, manufactured by Gibco) were ^{uniformly dispensed into a 6-well plate for culture with a constant number of 1 × 10 5} cells, and M254 (cat) (cat). # M-254-500 (manufactured by Gibco) was cultured in an incubator for 24 hours at 37 ° C. and 5% CO _2. Each of Compound 1 and Compound 2 is dissolved in DW at a concentration of 100 mM to make a concentrate, which is diluted with a medium to dilute it at a concentration of 20 μM, and then each dilution is made with 1 ml of the medium first in each well. After treating with 1 ml of each solution, the cells were cultured for 48 hours, and RNA was extracted from melanin cells using a RNeasy mini kit (cat # 74106, manufactured by Kiagen). The extracted RNA is cDNA-synthesized using a cDNA synthesis kit (cat # FSQ-201, manufactured by Toyo Boseki Co., Ltd.), and MITF (F: TAA CCT GTA CAA CAA CTC TCG ATC TCA; R: GTT GGC CTC AGT CCC AGT TC ), Tyrosinase (F: AGC ACC CCA CAA ATC CTA ACT TAC; R: ATG GCT GTT GTA CTC CTC CAA TC) Primer and POWER SYBR MASTER MIX (cat # 4367659) synthesized with Thermo Fisher Scientific The mixture was mixed at an appropriate ratio, and qPCR was performed using Quant Studio 3 equipment (cat # A28131, manufactured by Thermo Fisher Scientific). In addition, after the same experiment was performed and the culture was completed, the medium was removed, and melanocytes were separated from the plate using trypsin-EDTA (cat # LS-015-09, manufactured by WELGENE), and 1.5 ml was used. After transfer to a tube, the pellets were spin down to take a picture of the pellets, and the absorbance of the solution of the pellets dissolved in 1N NaOH was measured (wavelength: 400 nm, Epoch, manufactured by Biotech).

The results are shown in FIGS. 5 to 7.

As shown in FIGS. 5 to 7, it can be seen that the compounds 1 and 2 obtained in the above examples have almost no effect on the melanosome maturation activity in melanocytes.

(Example 6) Confirmation of increase in autophagy activity of keratinized cells and decrease in expression of endocytosis-related genes LC3 (light chain) for intracellular autophagy analysis by treatment with compound 1 obtained in the above example. 3; Autophagy marker) Western blotting (western blotting) was performed on the protein. Moreover, in order to analyze the change of the endocytosis-related gene obtained by the treatment of compound 2 obtained in the said Example, qPCR was performed on PAR2 and AAK1 (AP2 Associated Kinase1).

As a specific experimental method, compound 1 is treated into keratinized cells (HEKn, cat # C0015C, manufactured by Gibco) and lysed with a cell lysis buffer containing a proteolysis inhibitor. I let you. After fractionating the dissolved solution using a centrifuge, only the solution containing the protein was extracted. The extracted protein was quantified by the BCA (bicinchoninic acid) method. 20 μg of protein was separated by 10% SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane. The protein-transferred membrane is TBS-T (Tris-buffered saline / 0.05% twin-20) containing 5% skim milk (non-fat milk) to reduce non-specific binding. ) The primary antibody was reacted overnight (overnight) under the condition of 4 ° C. after blocking with the solution at room temperature for 1 hour, and then the secondary antibody was reacted at room temperature for 1 hour. A chemiluminescent system was used for visualization.

As a specific experimental method, keratinized cells treated with Compound 2 (HEKn, cat # C0015C, manufactured by Gibco) are RNA-extracted using an RNeasy mini kit (cat # 74106, manufactured by Chiagen), and cDNA synthesis is performed. After cDNA synthesis using a kit (cat # FSQ-201, manufactured by Toyo Boseki Co., Ltd.), PAR2 (F: CTC TCC TGC AGT GGC ACC AT; R: GAT GTG CCA TCA ACC TTA CCA A) primer and POWER SYBR MASTER Cat # 4367659 (manufactured by Thermo Fisher Scientific Co., Ltd.) was mixed with the synthesized cDNA in an appropriate ratio, and qPCR was performed with Quant Studio 3 equipment (cat # A28131, manufactured by Thermo Fisher Scientific Co., Ltd.).

The result is shown in FIG.

As shown in FIG. 8, it was confirmed that Compound 1 increased the production of LC3-II (mature autophagosome), and that Compound 2 decreased the expression of the PAR-2 gene. It was.

(Example 7) Confirmation of reduction in melanin deposition In order to analyze how the compounds 1 and 2 obtained in the above example affect melanin deposition, the degree of inhibition of deposition by melanin treatment was analyzed.

As a specific experimental method, human-derived keratinized cells (HEKn, cat # C0015C, manufactured by Gibco) were uniformly ^{dispensed into a 6-well plate for culture with a constant number of 1 × 10 5} cells, and EpiLife (registered trademark). , Cat # MEPI500CA, manufactured by Gibco), cultured in an incubator for 24 hours at 37 ° C. and 5% CO _{2 conditions.} Each of Compound 1 or Compound 2 obtained in the above example was dissolved at a concentration of 100 mM to prepare a concentrated solution, which was diluted with a medium to dilute it at a concentration of 20 μM, and then 1 ml of the medium was first placed in each well. After treating with 1 ml of each diluted solution in the same state, the cells were cultured for 48 hours, 10 mg / ml of melanin was added in 2 μl portions, and the cells were further cultured for 2 hours. After culturing, the medium was removed, cells were separated from the plate using trypsin-EDTA (cat # LS-015-09, manufactured by WELGENE), transferred to a 1.5 ml tube, and then spun down to pellet. The absorbance of the solution in which the pellet was dissolved in 1N NaOH was measured (wavelength: 400 nm, Epoch, manufactured by Biotech).

The result is shown in FIG.

As shown in FIG. 9, it was confirmed that Compound 1 or Compound 2 effectively suppressed melanin deposition on keratinized cells.

It was also found that melanin deposition was further suppressed when these were mixed as compared with the case where compound 1 or compound 2 was treated with a single compound.

(Example 8) Analysis on whether the melanin deposition inhibitory activity is dependent on the autophagy activity The melanin deposition inhibitory activity on keratinized cells by the treatment of Compound 1 and Compound 2 obtained in the above Example is autophagy. In order to analyze whether it is activity-dependent, ATG5, which is a protein required for autophagy activity, was knock-downed, and then the change in the deposition inhibitory activity due to melanin treatment was analyzed.

As a specific experimental method, ATG5 siRNA is mixed by a transfection reagent (RNAiMAX, cat # 13778-075, manufactured by Thermo Fisher Scientific), and then keratinized cells (HEKn, cat # C0015C, manufactured by Gibco). ). After 8 hours, the cells 1 and 2 were diluted with a supplement (supplement) and an antibiotic-free EpiLife (cat # MEPI500CA, manufactured by Gibco) to a final concentration of 10 μM, and the cells were further cultured for 64 hours. It was lysed using a cell lysis buffer containing a proteolytic inhibitor. After fractionating the dissolved solution using a centrifuge, only the solution containing the protein was extracted. The extracted protein was quantified by the BCA (bicinchoninic acid) method. 20 μg of protein was separated by 10% SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane. The protein-transferred membrane is TBS-T (Tris-buffered saline) containing 5% BSA (bovine serum albumin, Gendepot, Cat # A0100-010) to reduce non-specific binding. After blocking with a / 0.05% twin-20) solution at room temperature for 1 hour, the primary antibody was reacted overnight under the condition of 4 ° C., and then the secondary antibody was reacted at room temperature for 1 hour. I let you. A chemiluminescent system was used for visualization. In addition, in order to analyze the melanin deposition inhibitory activity, transfection and treatment of the compound were carried out in the same manner, and then 10 mg / ml of melanin was added in an amount of 2 μl each, and the cells were further cultured for 2 hours. After completion of the culture, the culture solution is removed, cells are separated from the plate using trypsin-EDTA (cat # LS-015-09, manufactured by WELGENE), transferred to a 1.5 ml tube, and then spin down (spin). Down), a photograph of the pellet was taken, and the absorbance of the solution in which the pellet was dissolved in 1N NaOH was measured (wavelength: 400 nm, Epoch, manufactured by Biotech).

The result is shown in FIG.

As shown in FIG. 10, it was confirmed that when ATG5, which is a protein required for autophagy activity, was knock-down, the amount of melanin in keratinized cells by Compound 1 and Compound 2 did not decrease. It was also confirmed that the expression activity of the autophagy proteins LC3-II and ATG5 by Compound 1 and Compound 2 was suppressed under the condition that ATG5 was knock-down. Therefore, it was found that the melanin deposition inhibitory activity of Compound 1 and Compound 2 is dependent on the autophagy activity.

(Example 9) Melanosome decomposition effect In order to confirm whether the melanosomes and autophagosomes are co-localized by the treatment of compound 1 and compound 2 obtained in the above example, analysis was performed with an electron microscope and a confocal microscope.

As a specific experimental method, a lysis buffer solution (Tris 20 mm, NaCl 150 mm, EDTA 1 mM, sucrose 250 mm in DW) was added to melanocytes (HEMn-DP, cat # C2025C, manufactured by Gibco) containing a high concentration of human-derived melanin. The cell membrane is broken by repeating thawing twice after adding and cooling, and centrifuging at 1000 xg for 5 minutes to obtain a supernatant. Fill the ultracentrifuge tube with Cushion Buffer (Tris 20 mm, NaCl 150 mm, EDTA 1 mM, sucrose 1.5 Min DW), add the same amount of supernatant, and then centrifuge at 25,000 rpm for 30 minutes at 4 ° C. And obtained melanosomes. The concentration of melanosomes was measured using synthetic melanin (cat # M8631, manufactured by Sigma-Aldrich) as a standard. Keratinized cells (HEKn, cat # C0015C, manufactured by Gibco) were ^{dispensed into 6 wells at 1 × 10 5} cells / well and cultured for 1 day. Compound 1 and compound 2 were treated with 10 μM each and cultured for 1 day, then separated melanosomes (5 μg / well) were added, and the cells were further cultured for 2 days. After completion of the culture, the culture solution is removed, cells are fixed using an electron microscope fixation solution, fluorescent labeling is performed with an antibody (Pmel17, LC3), and then melanosomes and autophagosomes are co-localized under a confocal microscope. I confirmed whether to do it. In addition, the appearance of organelles was observed in the cells of the same experiment with a transmission electron microscope.

The results are shown in FIGS. 11 and 12.

As shown in FIG. 11, it was confirmed by confocal microscopy that the autophagosomes increased by Compound 1 and Compound 2 were co-localized with melanosomes. In addition, unlike the control in which a large amount of melanosomes are present in keratinized cells, it was confirmed that when a mixture of compound 1 and compound 2 was treated, the melanosomes were rapidly decomposed and the amount thereof was significantly reduced.

Further, as shown in FIG. 12, when the mixture of compound 1 and compound 2 was treated, it was possible to confirm the phenomenon that 86% of all melanosomes were contained in autophagosomes by a transmission electron microscope.

(Example 10) Effect of suppressing melanosome deposition The compounds 1 and 2 obtained in the above example were treated and separated from the keratinocytes in order to analyze whether the deposition of melanosomes was suppressed in the keratinocytes. Melanosomes were treated and melanin content and melanosome indicators were analyzed.

As a specific experimental method, keratinized cells (HEKn, cat # C0015C, manufactured by Gibco) were ^{dispensed into 6 wells at 1 × 10 5} cells / well and cultured for 1 day, and then compound 1 and compound 2 were added to 10 μM. Each was treated for 1 day, and the separated melanosomes as in Example 9 were added (5 μg / well) and then cultured for another 2 days. After completion of the culture, a culture solution was obtained, cells were separated from the plate using trypsin-EDTA (cat # LS-015-09, manufactured by WELGENE), transferred to a 1.5 ml tube, and then spun down to the pellet. A photograph was taken and the absorbance of the solution in which the pellet was dissolved in 1N NaOH was measured (wavelength: 400 nm, Epoch, manufactured by Biotech).

In addition, in order to quantitatively analyze this, the amount of melanosomes in the culture medium and cell pellet was analyzed using synthetic melanin (cat # M8631, manufactured by Sigma-Aldrich) as a standard product. The obtained culture solution was freeze-dried, dissolved in 1N NaOH in the same manner as the pellets, and then the absorbance was measured (wavelength: 400 nm, Epoch, manufactured by Biotech).

The result is shown in FIG.

As shown in FIG. 13, it was confirmed that when the mixture of Compound 1 and Compound 2 was treated, the amount of melanosomes inside the keratinized cells decreased and the amount of melanosomes in the culture medium increased. Such a result means that the influx of melanosomes into keratinocytes was effectively suppressed by the treatment of the mixture of Compound 1 and Compound 2. Thereby, when the mixture of compound 1 and compound 2 is treated, pigmentation due to melanosomes can be significantly suppressed.

In addition, the mechanism for suppressing the pigmentation process of keratinocytes according to the present invention is shown in FIG. 1 below.

(Example 11) Melanin deposition inhibitory effect In order to analyze how compounds 1 and 2 obtained in the above example have a histological effect on melanin deposition inhibition, melanin was used using an artificial skin model. The degree of hyperpigmentation suppression was analyzed.

As a specific experimental method, an artificial skin model derived from a dark-skinned person (MelanoDerm ™, MEL-300-B, manufactured by Mattek) was used in a special medium (EPI-100-NMM-PRF) for 15 days at 37 ° C. The cells were cultured in an incubator under the condition of 5% CO _2. Compound 1 and Compound 2 obtained in the above example were treated on the surface of the artificial skin model at a concentration of 100 ppm by 25 μl once every 24 hours. The existing compound 1 and compound 2 at the time of treatment of compound 1 and compound 2 were removed before treatment, and the medium was also replaced every 24 hours. Photographs of the tissue surface were taken on the 1st, 9th, and 15th days during the progress of the experiment, and the degree of color change (L * value) of the tissue surface was analyzed using the results.

The result is shown in FIG.

As shown in FIG. 14, it was confirmed that the melanin pigmentation was statistically significantly reduced in the group treated with the mixture of Compound 1 and Compound 2 as compared with the control.

(Example 12) Melanin-degrading effect In order to analyze the melanin-degrading effect of Compound 1 and Compound 2 obtained in the above-mentioned Example, the melanin content was analyzed using an artificial skin model.

As a specific experimental method, an artificial skin model was cultured as described in Example 11. Tissues were obtained on day 15 and fixed in 4% phosphate buffered formalin for 24 hours. The fixed tissue was washed with flowing water for 12 hours, dehydrated in descending order of alcohol concentration, replaced with xylene, and embedded in paraffin. The paraffin-embedded tissue was sliced to a thickness of 5 μm, stained with melanin by the Fontana Masson staining method, and then used for analysis. The photographs of all tissues and the photographs of the stratum corneum were analyzed and compared with imageJ, respectively.

The result is shown in FIG.

As shown in FIG. 15, it was confirmed that melanin was statistically significantly reduced in the group treated with the mixture of compound 1 and compound 2 as compared with the control. In addition, it was confirmed that the amount of melanin ascending to the stratum corneum was 43.25%, which was statistically significantly reduced as compared with the control.

From the above results, it was confirmed that the compound according to the present invention can effectively suppress the influx of melanosomes by effectively inducing melanosome degradation by autophagy activation and promoting PAR-2 suppression. did.

Specifically, compound 1 induces autophagy activity of melanocytes to effectively decompose melanosomes as a melanosomes degradation-inducing compound by autophagy activation, and compound 2 suppresses melanosomes influx by suppressing endocytosis. As an inducing compound, it was confirmed that it exerts an outstanding effect on the role of blocking melanosomes from entering keratinocytes. As a result, it is expected that melanin in keratinocytes will be reduced and it can be used very effectively for alleviation, treatment and prevention of skin pigmentation. In particular, when the compounds represented by the chemical formulas 2 and 3 according to the present invention are mixed and used, not only the whitening effect but also the excellent effect for the improvement, prevention and treatment of the disease due to the excessive deposition of the melanin pigment is realized. Can be done.

(Examples 13 to 14 and Comparative Example 1) Production of cosmetic composition for skin whitening The ingredients and contents shown in Table 2 below are put into an emulsification reactor and heated to 75 ° C. with mixing and stirring. It was emulsified. After that, when the emulsification was completed, the mixture was stirred with a stirrer and gradually cooled to room temperature to produce a skin whitening cosmetic composition (o / w cream).

(Example 15) Whitening effect A clinical test was conducted to investigate the whitening effect of the skin whitening cosmetic composition of the example produced by the above method.

As a specific experimental method, UV exposure induces darkening of the skin of 20 adult men and women (15 women and 5 men) with an average age of 37 years (36.57 ± 5.90) with hyperpigmentation. After that, as shown in Table 2, high-concentration and low-concentration-containing creams containing compound 1 and compound 2 (test sample) and creams not containing compound 1 and compound 2 (Comparative Example 1, control). Sample) was applied twice a day. After applying each cream, measure according to the manual of the instrument using Meter and Chromameter at the 4th and 8th weeks, and the measured values are measured according to the manual of the instrument, and the measured values are measured by ANOVA for the change due to the evaluation time after the normality test. By paired t-test, the significance of the test sample / control sample before and after sample application and weekly discrimination was confirmed with a hypothetical mean difference of 5% (P <0.05). The statistical analysis program used was IBM SPSS version 18.0.

The results are shown in Tables 3 and 4 below.

As shown in Tables 3 and 4, the melanin pigment in the group using the cosmetic composition containing the melanosomes decomposition-inducing compound by autophagy activation of the present invention and the melanosomes inflow suppression-inducing compound by suppressing endocytosis. It was confirmed that the deposition and the change in skin brightness were improved. In addition, it was confirmed that the improvement rate of melanin pigmentation and changes in skin brightness exhibited more improved effects as the usage time elapsed.

As described above, the present invention has been described by the specified matters and limited examples, which are provided for facilitating a more general understanding of the present invention. Is not limited to the above-described embodiment, and any person who has ordinary knowledge in the field to which the present invention belongs can make various modifications and modifications from the description.

Therefore, the idea of the present invention should not be limited to the above-described embodiment, and not only the claims described later, but also anything having a modification equal to or equivalent to the claims of the present invention, etc. , It can be said that it belongs to the category of the idea of the present invention.

Claims (17)

A compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

In the chemical formula 1,
AA is an amino acid residue containing substituted or unsubstituted proline. The first aspect of claim 1, wherein the AA is an amino acid residue comprising proline substituted or unsubstituted with one or more substituents selected from _{hydroxy, C 1-10} alkoxy, C _{1-10 alkylcarbonyl and carboxy.} Compound or pharmaceutically acceptable salt thereof. The chemical formula 1 is the compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is represented by the following chemical formula 2.

In the chemical formula 2,
R ₁ is a C _1-10 alkyl carbonyl;
R ₂ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. The compound represented by the chemical formula 1 is the compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is represented by the following chemical formula 3.

In the chemical formula 3,
R ₃ is hydroxy or C _1-10 alkyl;
R ₄ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. A cosmetic composition for skin whitening containing the compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In the chemical formula 1,
AA is an amino acid residue containing substituted or unsubstituted proline. The active ingredient is
The cosmetic composition for skin whitening according to claim 5, which is a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt (A) thereof.

In the chemical formula 2,
R ₁ is a C _1-10 alkyl carbonyl;
R ₂ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. The active ingredient is
The cosmetic composition for skin whitening according to claim 5, which is a compound represented by the following chemical formula 3 or a pharmaceutically acceptable salt (B) thereof.

In the chemical formula 3,
R ₃ is hydroxy or C _1-10 alkyl;
R ₄ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. The active ingredient is
A claim which is a mixture of the compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt (A) thereof and the compound represented by the following chemical formula 3 or a pharmaceutically acceptable salt (B) thereof. 5. The cosmetic composition for skin whitening according to 5.

In the chemical formulas 2 and 3,
R ₁ is a C _1-10 alkyl carbonyl;
R ₃ is hydroxy or C _1-10 alkyl;
R ₂ and R ₄ are independently hydrogen, hydroxy, C _1-10 alkoxy, C _1-10 alkylcarbonyl or carboxy, respectively. The active ingredient is
The skin whitening cosmetic composition according to claim 8, which is mixed in a weight ratio of 1: 0.1 to 1: 1 (A: B). The skin whitening cosmetic composition according to any one of claims 5 to 9, which contains 0.0001 to 10% by weight of the active ingredient with respect to the total weight. A pharmaceutical composition for improving or treating a melanin pigment hyperdeposition disease, which comprises a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In the chemical formula 1,
AA is an amino acid residue containing substituted or unsubstituted proline. The active ingredient is
The pharmaceutical composition for ameliorating or treating an over-deposition disease of a melanin pigment according to claim 11, which is a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt (A) thereof.

In the chemical formula 2,
R ₁ is a C _1-10 alkyl carbonyl;
R ₂ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. The active ingredient is
The pharmaceutical composition for ameliorating or treating a hyperdeposited disease of a melanin pigment according to claim 11, which is a compound represented by the following chemical formula 3 or a pharmaceutically acceptable salt (B) thereof.

In the chemical formula 3,
R ₃ is hydroxy or C _1-10 alkyl;
R ₄ is hydrogen, _{hydroxy, C 1-10 alkoxy, C 1-10} alkylcarbonyl or carboxy. The active ingredient is
It is a mixture of the compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt (A') thereof and the compound represented by the following chemical formula 3 or a pharmaceutically acceptable salt thereof (B'). The pharmaceutical composition for improving or treating an over-deposited disease of melanin pigment according to claim 11.

In the chemical formulas 2 and 3,
R ₁ is a C _1-10 alkyl carbonyl;
R ₃ is hydroxy or C _1-10 alkyl;
R ₂ and R ₄ are independently hydrogen, hydroxy, C _1-10 alkoxy, C _1-10 alkylcarbonyl or carboxy, respectively. The active ingredient is
The pharmaceutical composition for ameliorating or treating an over-deposited disease of melanin pigment according to claim 14, which is mixed in a weight ratio of 1: 0.1 to 1: 1 (A': B'). The melanin pigment excess deposition disease is
With freckles, senile pigmented spots, chloasma, spots, brown or black spots, sun-induced pigmented spots, blue melanosis, excessive pigmentation after drug use, gestational brown spots or excessive pigmentation after inflammation The pharmaceutical composition for improving or treating the hyperpigmentation disease of melanin pigment according to claim 11. The pharmaceutical composition for improving or treating an over-deposited disease of melanin pigment according to any one of claims 11 to 16, which contains 0.0001 to 10% by weight of the active ingredient based on the total weight.

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