JP6883090B2 - A composition for skin whitening containing 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide. - Google Patents

Description

This specification describes skin containing 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide. It relates to a composition for whitening, and specifically, it relates to a composition containing a substance that regulates the production and deposition of melanin and exerts a skin whitening effect.

Melanin is a phenolic biopolymer in the form of a complex of black pigment and protein that causes browning or browning that occurs when the cut surface of apples, potatoes and bananas is exposed to the air. It is observed in the outer skin, skin, hair, and eyes of animals. When melanin is overproduced and deposited on the skin, stains and freckles are formed, which is directly linked to skin whitening, and melanin also promotes skin aging and may induce skin cancer.

Melanocyte stimulating hormone (MSH) is secreted by ultraviolet rays, inflammation, hormones, etc., and the MSH reacts with the receptor to improve cAMP in melanin-forming cells to synthesize melanin, and the synthesized melanin Is secreted to the outside of melanin-forming cells and plays a role in protecting the skin from ultraviolet rays and the like. The synthesis of melanin is mainly regulated by α-MSH, and MITF, TYR, TRP1, TRP2 and the like are known as proteins involved in the synthesis of melanin.

CHEN, J.K. et al., Small Molecule Modulation of Smoothened Activity., Proceedings of the National Academy of Science, 2002, vol. 99, no. 22, pages 14071-14076 YANG, H. et al., Converse Conformational Control of Smoothened Activity by Structurally Related Small Molecules., Journal of Biological Chemistry, 2009, vol. 284, no. 31, pages 20876-20884 MAK, S.K. et al., Small Molecules Greatly Improve Conversion of Human-induced Pluripotent Stem Cells to the Neuronal Lineage, Stem Cells International, 2012, vol. 2012, thesis no. 140427, internal pages 1-12 FRANK-KAMENETSKY, M. et al., Small-molecule Modulations of Hedgehog Signaling: Identification and characterization of Smoothened Agonists and Antagonists., Journal of Biology, 2002, vol. 1, no. 2, thesis no. 10, internal pages 1 -19

The present invention, in one aspect, is 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide. To provide a composition for skin whitening containing, an optical or stereoisomer thereof, an acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient, to develop a field related to skin whitening, and to develop skin. Its purpose is to meet the demands of whitening-related consumers.

The present invention, in one aspect, is 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide. , An optical or stereoisomer thereof, an acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

In another aspect of the present invention, the 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2- Carboxamide may be one that suppresses the production and deposition of melanin.

Further, in another aspect of the present invention, the composition may be a pharmaceutical composition or a cosmetic composition.

The present invention, in one aspect, is 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide. (3-Chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide), its optical or stereoisomer , A skin whitening method comprising administering to an individual an acceptable salt thereof, a hydrate thereof, or a solvate thereof.

In addition, in another aspect, the present invention comprises 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) as an active ingredient for use in skin whitening applications. ) Phenyl] methyl] benzo [b] thiophen-2-carboxamide, an optical or stereoisomer thereof, an acceptable salt thereof, a hydrate thereof, or a solvate thereof. In addition, as an active ingredient for skin whitening, 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene- Provided are non-therapeutic cosmetic uses of 2-carboxamide, an optical or stereoisomer thereof, an acceptable salt thereof, a hydrate thereof, or a solvate thereof.

In addition, in another aspect, the present invention comprises 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) for use in the production of skin whitening compositions. ) Phenyl] methyl] benzo [b] thiophen-2-carboxamide, an optical or stereoisomer thereof, an acceptable salt thereof, a hydrate thereof, or a solvate thereof.

The present invention, in one aspect, is 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide. Opening up new fields related to skin whitening by discovering and providing new skin whitening compositions containing, its optical or stereoisomer, its acceptable salt, its hydrate, or its solvate as an active ingredient. And can contribute to the expansion of the market.

B16F1 cells with α-MSH, 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide, It is a figure which showed the change of the content of melanin when arbutin was treated. B16F1 cells with α-MSH, 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide, It is a figure which confirmed the influence on the expression of tyrosinase, TRP1 when arbutin was treated. After treating the skin model with 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide It is the figure which confirmed the change of brightness. After treating the skin model with 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide It is the figure which evaluated the change of the brightness by the chromaticity meter.

As used herein, the term "skin" means a tissue covering the body surface of an animal, and is a broadest concept including not only the tissue covering the body surface such as the face or the body but also the scalp and hair.

In the present specification, "3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide" is used. , Smo agonist, Smoothered agent, Smo agent, or SAG. The 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide is a Hedgehog signaling pathway. It may contain a substance that induces the expression of a Smoothened protein that plays a major role in (hedgehog signaling pathway).

As used herein, "isomers" are particularly pure optical isomers (eg, essentially pure isomers), essentially pure steric isomers or mixtures thereof, essentially pure. Essentially pure diastereomers, as well as steric isomers, conformation isomers (ie, isomers that differ only in their angle of one or more chemical bonds), position isomers. (In particular, tautomers), or geometric isomers (eg, cis-trans isomers).

As used herein, "essentially pure" is, for example, an enantiomer, a stereoisomer, or a diastereomer when used in connection with an enantiomer, a stereoisomer, or a diastereomer. About 90% or more, preferably about 95% or more, more preferably about 97% or more, or about 98% or more, still more preferably about 99% or more, most preferably about 99% or more. It means that it is present at 5% or more (w / w).

As used herein, "acceptable" can be used in animals, more specifically in humans, by avoiding significant toxicity when used in normal or pharmaceutical doses. Can be approved or approved by the government or an equivalent regulatory body, or listed in the Food Code, Health Function Food Standards, or General Pharmacopoeia, or other general literature. It means that it is certified as described in.

As used herein, the term "acceptable salt" means a salt according to one aspect of the invention that is usually or pharmaceutically acceptable and has the preferred activity of the parent compound. Is the salt formed from (1) inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitrate, phosphoric acid and the like; or acetic acid, propionic acid, hexanic acid, cyclopentenepropionic acid, glycolic acid, pyruvate, Lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2,2 , 2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, laurylsulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, mucon It may include an acid addition salt formed from an organic acid such as an acid; or (2) a salt formed when the acidic protons present in the parent compound are replaced.

As used herein, the term "hydrate" means a compound to which water is bound, and is a broad concept including an inclusion compound having no chemical binding force between water and a mixture.

As used herein, the term "solvate" means a higher-order compound formed between a molecule or ion of a solute and a molecule or ion of a solvent.

The present invention, in one aspect, is 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide. , An optical or stereoisomer thereof, an acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

As an embodiment, the 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide (3) -Chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide) is a chlorobenzothiophene compound. It _{may contain C 28} H ₂₈ ClN ₃ OS, and may be represented by the following chemical formula.

As another embodiment, the compound may be obtained by synthesis, may be obtained by processing another substance, or may be derived from an organism, a microorganism, or the like.

As another embodiment, the 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide , It may suppress the production or deposition of melanin.

According to another aspect of the present invention, the 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2 -Carboxamide may enhance the expression of tyrosinase or TRP1 (Tyrosinase-Related productin 1). Since the expression of tyrosinase or TRP1 after the treatment of the compound is lower than that before the treatment of the compound, it can be seen that the compound can be used as an active ingredient of the skin whitening composition. In addition, when the compound is treated, the number of cells expressing tyrosinase or TRP1 may be reduced, and it can be seen that the compound can be used as an active ingredient of a skin whitening composition.

Melanin is observed from the outer skin hair, skin, head, eyes, etc. of animals, and when melanin is overproduced, it deposits on the skin and forms spots, freckles, etc., and also accelerates skin aging. , May also induce skin cancer. Diseases or symptoms due to overproduction of melanin include stains, moles, senile stains, fine dust, epidermal melanotic lesions, cafe's au light scales, moles, and moles of Becker. (Becker's Nevus), Mole (Nevus Spils), Mole (Lentiines), Black spots, Dermal melanotic lesions, Mongolian spot, Ota mother's spot (Nevus) Two-sided bilateral Ota mole-like pigmented mole (Acquired bilateral nevus of Ota-like moles), Ito mole (Nevus of Ito), blue mole (Blue nevus), melanogenic cell mole (Melanocytic) Junctional nevus, Compound nevus, Intradermal nevus, Mole nevus, Congenital melanin cell spicy nevus, Congenital nevocatic nevus Spots (Spitz nevus), dysplastic nevus, melanoma (Melanoma), malignant melanoma, superficial spreading melanoma, terminal melanoma (Acral lentiginous melanoma), nodular melanoma, pigmented basal cell carcinoma, pigmented dermatofibromas, pigmented mole cyst, pigmented skin cyst (dermatofibromas) It may be one or more selected from the group consisting of moleid), ultraviolet pigmentation, drug pigmentation, post-inflammatory pigmentation, and pigmentation and pigmented keratoacanthomas resulting from dermatitis.

As an embodiment of the present invention, the 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2- Carboxamide suppresses the production and accumulation of melanin from spots, freckles, black spots, nevus, melanoma, UV pigmentation, drug-induced hyperpigmentation, post-inflammatory hyperpigmentation, and pigmentation resulting from dermatitis. One or more selected from the group may be prevented, ameliorated or treated.

In one aspect of the present invention, the process of treating melanin-forming cells with a substance that induces melanin formation to form melanin is 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [. It was confirmed that it was suppressed by [3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide.

Based on these results, the present inventors have identified 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene. It has been found that -2-carboxamide can be used as a skin whitening substance. Using the efficacy of 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide. It is also possible to prevent, improve, and treat the above-mentioned diseases and symptoms related to melanin.

According to yet another aspect of the invention, 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [in the composition. b] The content of thiophen-2-carboxamide, an isomer thereof, an acceptable salt thereof, a hydrate thereof, or a solvate thereof is 0.0001% by mass to 20% by mass based on the total weight of the composition. It may be in the range of. In one embodiment, the content is 0.0001% by mass or more, 0.0005% by mass or more, 0.001% by mass or more, 0.005% by mass or more, 0.01% by mass with respect to the total weight of the composition. % Or more, 0.05% by mass or more, 0.1% by mass or more, 0.3% by mass or more, 0.5% by mass or more, 0.8% by mass or more, 1% by mass or more, 3% by mass or more, 5% by mass % Or more, 8% by mass or more, 10% by mass or more, 12% by mass or more, 15% by mass or more, or 18% by mass or more. The content is 20% by mass or less, 18% by mass or less, 15% by mass or less, 12% by mass or less, 10% by mass or less, 8% by mass or less, 5% by mass or less, based on the total weight of the composition. 3% by mass or less, 1% by mass or less, 0.8% by mass or less, 0.5% by mass or less, 0.3% by mass or less, 0.1% by mass or less, 0.05% by mass or less, 0.01% by mass Hereinafter, it may be 0.005% by mass or less, 0.001% by mass or less, or 0.0005% by mass or less.

According to yet another aspect of the present invention, the 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene- The dose of 2-carboxamide, an isomer thereof, an acceptable salt thereof, a hydrate thereof, or a solvate thereof may be in the range of 0.0001 mg / kg / day to 20 mg / kg / day. In one embodiment, the doses are 0.0001 mg / kg / day or higher, 0.0005 mg / kg / day or higher, 0.001 mg / kg / day or higher, 0.005 mg / kg / day or higher, 0.01 mg / day or higher. kg / day or more, 0.05 mg / kg / day or more, 0.1 mg / kg / day or more, 0.5 mg / kg / day or more, 0.8 mg / kg / day or more, 1 mg / kg / day or more, 2 mg / day or more kg / day or more, 3 mg / kg / day or more, 5 mg / kg / day or more, 8 mg / kg / day or more, 10 mg / kg / day or more, 12 mg / kg / day or more, 15 mg / kg / day or more, or 18 mg / day It may be kg / day or more. The doses are 20 mg / kg / day or less, 18 mg / kg / day or less, 15 mg / kg / day or less, 12 mg / kg / day or less, 10 mg / kg / day or less, 8 mg / kg / day or less, 5 mg. / Kg / day or less, 3 mg / kg / day or less, 2 mg / kg / day or less, 1 mg / kg / day or less, 0.8 mg / kg / day or less, 0.5 mg / kg / day or less, 0.1 mg / kg / Day or less, 0.05 mg / kg / day or less, 0.01 mg / kg / day or less, 0.005 mg / kg / day or less, 0.001 mg / kg / day or less, or 0.0005 mg / kg / day or less It may be there.

According to one aspect of the present invention, the composition may be a skin whitening composition which is a pharmaceutical composition.

The pharmaceutical composition is a preservative, a preservative, a stabilizer, a wettable powder or an emulsion promoter, a pharmacological aid such as a salt and / or a buffer for osmoregulation, and other therapeutically useful. Substances may be further contained and may be emulsified in the form of various oral or parenteral administrations according to conventional methods.

Examples of the oral administration agent include tablets, pills, hard and soft capsules, liquids, suspending agents, emulsifiers, syrups, powders, powders, fine granules, granules, pellets and the like. In addition to the active ingredient, the form is a surfactant, a diluent (eg lactose, dextrose, syrup, mannitol, sorbitol, cellulose and glycine), a lubricant (eg silica, talc, stearic acid and its magnesium or calcium salt). , And polyethylene glycol). The tablets may further contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose and polyvinylpyrrolidin, and optionally a disrupting agent such as starch, agar, alginic acid or a sodium salt thereof. , Absorbents, colorants, flavors, and pharmaceutical additives such as sweeteners. The tablets may be produced by conventional mixing, granulation or coating methods.

The form of parenteral administration may be a transdermal dosage form, for example, an injection, an infusion, an ointment, a lotion, a gel, a cream, a spray, a suspension, an emulsion, a suppository, or a patch. However, the dosage form is not limited to these.

The pharmaceutical composition may be administered parenterally, rectal, topically, transdermally, subcutaneously or the like.

The determination of the dose of the active ingredient is within the standard of a normal technician, and the daily dose of the drug varies depending on the degree of progression of the subject to be administered, the time of onset, age, health condition, complications, etc. It can vary depending on various factors.

The pharmaceutical composition may be an external preparation for skin, and the external preparation for skin is a general term for compositions that can contain any of those applied to the outer skin, and is a pharmaceutical product or quasi-drug in various dosage forms. Can be included in this.

According to yet another aspect of the present invention, the composition may be a skin whitening composition which is a cosmetic composition.

The cosmetic composition may further contain functional additives and ingredients contained in general cosmetic compositions. The functional additive may contain a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, sphingolipids and seaweed extracts. Other ingredients included include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjustments. Examples include agents, alcohols, pigments, fragrances, blood circulation promoters, cold sensation agents, antiseptic agents, purified water and the like.

The dosage form of the cosmetic composition is not particularly limited, and the cosmetic composition may be appropriately selected depending on the intended purpose. For example, Skin Lotion, Skin Softener, Skin Toner, Astringent, Lotion, Milk Lotion, Moisture Lotion, Nutrition Lotion, Massage Cream, Nutrition Cream, Moisture Cream, Hand Cream, Foundation, Essence, Nutrition Essence, Pack, Soap, Cleansing Foam, It may be produced in any one or more dosage forms selected from the group consisting of cleansing lotions, cleansing creams, body lotions, and body cleansers, but is not limited thereto.

When the dosage form according to one aspect of the present invention is a paste, cream, or gel, the carrier components include animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicon, bentonite, silica, and the like. Starch, zinc oxide, or the like may be used.

When the dosage form according to one aspect of the present invention is powder or spray, propane, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as the carrier component, and particularly when it is a spray. May further include propellants such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

When the dosage form according to one aspect of the present invention is a solution or an emulsion, a solvent, a solvent admixture, or an emulsion agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, etc. There are ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol, or fatty acid esters of sorbitan.

When the dosage form according to one aspect of the present invention is a suspension, the carrier components include water, ethanol, or a liquid diluent such as propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and poly. Suspensions such as oxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxylide, bentonite, agar, tragacant and the like may be used.

When the dosage form according to one aspect of the present invention is a surfactant-containing cleansing, as a carrier component, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinate monoester, an acetylonic acid, an imidazolium derivative, Methyl tauret, sarcosinate, fatty acid amide ether sulfate, alkylamide betaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, ethoxylated glycerol fatty acid ester and the like may be used.

Hereinafter, the configuration and effect of one aspect of the present invention will be described more specifically through Examples and Experimental Examples. It should be noted that the following examples are merely exemplified for the purpose of assisting the understanding of the present invention, and the scope and scope of the present invention are not limited by them.

[Example 1]
Preparation of cell and skin model Normal human epidermal melanocytes (Normal human epidermal melanocytes, NHEMs, Cascade Biologics, Portland, OR, USA) were obtained and melanocytes derived from C57BL / 6J (black, a / a) mice. A melan-A cell line was obtained from Dr. Dorothy C. Bennett (St. George's Hospital Medical School, London, UK). The B16F1 mouse melanoma cell line was obtained from ATCC (Manassas, VA, USA).

Normal human epidermal melanocytes were maintained in M-254 medium under Human Melanocyte Growth Supplements, HMGS (Cascade Biologics, Inc., Mansfield, UK). Melan-A cells were found in RPMI 1640 medium with 10% (v / v) fetal bovine serum, 1% (v / v) penicillin-streptomycin, and 0.2 μM phorbol 12-mylstart 13-acetylate 13-. It was maintained under accept). B16F1 mouse melanoma cell lines were cultured in DMEM supplemented with 10% (v / v) fetal bovine serum and 1% (v / v) penicillin-streptomycin.

After obtaining the three-dimensional human skin substitute material (Thre-dimensional human skin substitut, MelanoDerm ™, MEL-312-B, MatTek, Seoul, Korea), KGF (Keratinocyte Growth Factor (Keratinocyte Growth Factor) It was maintained in EPI-100-NMM-113 medium optimized for Factor) and α-MSH (alpha-melanocyte stimulating homones) according to the manufacturer's instructions.

The change in brightness (ΔL) index of the skin model is calculated from the L index (L value, lightness index) measured by a chromaticity meter (colorimeter, CR-300, Konica Minolta, Tokyo, Japan), and ΔL. = Calculated as L (treated skin) -L (control group skin) on day 13 or 20. Increased ΔL index indicates compound-induced pigment under-deposition in the skin (Lee et al., 2013).

Reagent 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide "SAG" Obtained from Calbiochem (San Diego, CA, USA). α-MSH, arbutin, and kojic acid were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Melanin assay cells were treated with trypsin / EDTA, centrifuged at 1000 xg for 5 minutes and washed twice with PBS. The final cell pellet in the tube was photographed and the cell pellet was dissolved in 1N NaOH. The homogenized cell extract was transferred to a 96-well plate and the relative melanin content at absorbance 405 nm was measured using an ELISA plate reader.

Western blot analysis All lysates were 2 x Laemmli sample buffer, 62.5 mM Tris-HCl, pH 6.8, 25% [v / v] glycerol, 2% [w / v] SDS, 5 Prepared using% [v / v] β-mercaptoethanolol and 0.01% [w / v] bromophenol blue) (Bio-Rad, Hercules, CA, USA). All cellular proteins were measured using Bradford solution (Bio-Rad). The sample was then separated by SDS-PAGE and transferred to PVDF membrane (Bio-Rad). After blocking with 4% (w / v) skim milk in TBST (25 mM Tris, 140 mM NaCl, and 0.05% [v / v] Tween® 20), the membrane is identified as 1 It was cultured overnight with the next antibody.

Anti-actin antibody (anti-actin antibody, MAB1501, 1: 10,000) was obtained from Millipore (Temecula, CA, USA) and anti-αPEP7 (tyrosinase) antibody (anti-αPEP7 (tyrosinase) antibody, 1: 1000). And anti-αPEP1 (TRP1) antibody (anti-αPEP1 (TRP1) antibody, 1: 1000) was obtained from VJ Hairing (NIH, Bethesda, MD). For protein detection, the membrane was cultured with HRP-conjugated secondary antibody and the signal was signaled using SuperSignal® West Dura HRP Detection Kit (Pierce, Rockford, IL, USA). Was measured.

Statistical analysis Each data was obtained by performing at least 3 independent experiments and was expressed as mean ± SE (standard error). Statistical evaluation of the results was performed using one-way analysis of variance (1-way ANOVA) (*: p <0.05, **: p <0.01).

[Experimental Example 1] Regulation of melanin formation 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide Is the substance represented by the following chemical formula 1.

B16F1 cells were pretreated with α-MSH (1 μM) and 24 hours later, 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl. ] Benzo [b] thiophene-2-carboxamide (5 μM) or arbutin (500 μM) was treated. After 24 hours, the cell melanin content and the expression levels of tyrosinase and TRP1 protein were analyzed by Western blotting. As a result, the compound suppressed the production of melanin in B16F1 cells treated and cultured with α-MSH (Fig. 1). The compound also reduced the expression levels of tyrosinase and TRP1 protein (Fig. 2).

[Experimental Example 2] Artificial skin experiment A three-dimensional human skin substitute material (recombinant epidermal model containing normal human epidermal corneocytes and NHEMs) was used to experiment with the regulation of melanin production.

Specifically, 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide (5 μM), Kojic acid (KA, 5 mM) was administered to a three-dimensional human skin substitute once every three days for 13 and 20 days to evaluate its effect on pigmentation. The pigmentation was evaluated by a colorimeter, and in FIG. 4, ΔL is a value obtained by calculating the difference in brightness between the control group and the substance treatment group.

As a result, the compound reduced skin pigmentation (FIGS. 3 and 4). From this, it was confirmed that the compound regulates melanin formation and pigmentation even in a three-dimensional human skin substitute.

Hereinafter, examples of dosage forms of the composition according to one aspect of the present invention will be described, but application to various other dosage forms is also possible, and these are not intended to limit the present invention, simply. This is for a specific explanation.

[Dosage Form Example 1] Soft Capsules 3-Chloro-N- [Trans-4- (Methylamino) Cyclohexyl] -N- [[3- (4-Pyridinyl) Phenyl] Methyl] Benzo [b] Thiophen-2- 20 mg of carboxamide, 80 to 140 mg of L-carnitine, 180 mg of soybean oil, 2 mg of palm oil, 8 mg of hydrogenated vegetable oil, 4 mg of yellow wax and 6 mg of lecithin were mixed and filled into one capsule according to a usual method to prepare a soft capsule. ..

[Dosage Form Example 2] Tablets 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide 20 mg , 200 mg of galactooligosaccharide, 60 mg of lactose and 140 mg of maltose were mixed and granulated using a fluidized layer dryer, and then 6 mg of sugar ester was added and tableted with a tableting machine to produce tablets. ..

[Dosage Form Example 3] Granules 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide 20 mg, 250 mg of anhydrous crystalline glucose and 550 mg of starch were mixed, formed into granules using a fluidized bed granulator, and then packed in a packet to produce granules.

[Formation Example 4] Drinking agent 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide After mixing 20 mg, 10 g of glucose, 0.6 g of citric acid, and 25 g of liquid oligosaccharide, 300 ml of purified water is added, and each bottle is filled with 200 ml. After filling the bottle, it was sterilized at 130 ° C. for 4 to 5 seconds to produce a drink.

[Dosage Form Example 5] Injection 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide An injection was prepared according to a usual method using 50 mg, an appropriate amount of sterile distilled water for injection, and an appropriate amount of a pH adjuster.

[Dosage form example 6] Soft lotion (skin lotion)
3-Chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide 3.0% by mass, L- Ascorbic acid-2-magnesium phosphate salt 1.00% by mass, water-soluble collagen (1% aqueous solution) 1.00% by mass, sodium citrate 0.10% by mass, citric acid 0.05% by mass, licorice extract 0. A soft cosmetic solution (skin lotion) was produced using 20% by mass, 1,3-butylene glycol 3.00% by mass, and purified water as the remaining amount.

[Dosage Form Example 7] Cream 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide 3 .00% by mass, polyethylene glycol monostearate 2.00% by mass, self-emulsifying glycerin monostearate 5.00% by mass, propylene glycol 4.00% by mass, squalene 6.00% by mass, tri2-ethylhexaneglyceryl A creamy preparation was produced using 6.00% by mass, 1.00% by mass of sphingo glycolipid, 7.00% by mass of 1,3-butylene glycol, 5.00% by mass of beeswax, and purified water as the remaining amount.

[Formation Example 8] Pack 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide 3 0.00% by mass, polyvinyl alcohol 13.00% by mass, L-ascorbic acid-2-maganoate phosphate 1.00% by mass, lauroyl hydroxyproline 1.00% by mass, water-soluble collagen (1% aqueous solution) 2.00 A pack was produced using purified water in an amount of% by mass, 1,3-butylene glycol 3.00% by mass, ethanol 5.00% by mass, and the remaining amount.

[Dosage Form Example 9] Health food A health food was produced according to a usual method with the composition shown in the table below.

The composition ratio of the vitamin and the inorganic mixture is a mixture of ingredients that are relatively suitable for health foods as an example, but the composition ratio may be arbitrarily modified and according to a normal health food production method. After mixing the above components, it may be used in the production of a composition of a health food according to a usual method.

[Dosage form example 10] Health drink

As shown in Table 2, the remaining amount of purified water is added so as to have a total volume of 900 ml, the components are mixed according to a normal method for producing a healthy beverage, and then stirred and heated at 85 ° C. for about 1 hour to prepare the mixture. The filtrate obtained by filtering the resulting solution was placed in a sterilized 2 liter container, sterilized by sealing, and then refrigerated to produce a healthy beverage.

Although a specific part of the content of the present invention has been described in detail above, such a specific description is merely a preferred embodiment for a person having ordinary knowledge in the art, and thereby the scope of the present invention. It will be clear that is not limited. Therefore, it can be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (8)

3-Chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide (3-Chloro-N- [ trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] bento [b] thiophene-2-carboxamide), its optical or steric isomer, its acceptable salt, its acceptable salt. A composition for skin whitening containing a hydrate or a solvate thereof as an active ingredient. The 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide produces or deposits melanin. The composition for skin whitening according to claim 1, which suppresses the above-mentioned. The 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophene-2-carboxamide is tyrosinase or The composition for skin whitening according to claim 1, which enhances the expression of TRP1 (Tyrosinase-Related product 1). The 3-chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide is a stain, freckle, black color. Claim to prevent, ameliorate or treat one or more selected from the group consisting of spots, melasma, melanoma, hyperpigmentation due to ultraviolet rays, pigmentation due to drugs, post-inflammatory hyperpigmentation, and pigmentation caused by dermatitis. The composition for skin whitening according to 1. 3-Chloro-N- [trans-4- (methylamino) cyclohexyl] -N-[[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide in the composition, the opposite sex thereof. The first aspect of claim 1, wherein the content of the isomer, an acceptable salt thereof, a hydrate thereof, or a solvate thereof is in the range of 0.0001% by mass to 20% by mass with respect to the total weight of the composition. Composition for skin whitening. 3-Chloro-N- [trans-4- (methylamino) cyclohexyl] -N- [[3- (4-pyridinyl) phenyl] methyl] benzo [b] thiophen-2-carboxamide, its isomer, and its tolerance The skin whitening composition according to claim 1, wherein the possible dose of the salt, its hydrate, or its solvate is in the range of 0.0001 mg / kg / day to 20 mg / kg / day. The composition for skin whitening according to any one of claims 1 to 6 , wherein the composition is a pharmaceutical composition. The composition for skin whitening according to any one of claims 1 to 6 , wherein the composition is a cosmetic composition.

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