JP6952104B2 - Composition for enhancing melanin containing 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylidene) -β-oxo-benzenepropanenitrile - Google Patents

Description

This specification describes melanin-lowering properties for blackening hair containing 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile. It relates to a composition for preventing a disease or for treating a melanin-lowering disease.

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, and it is directly linked to blackening of hair. Melanin also promotes skin or hair 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 melanocyte-forming cells and plays a role in protecting the skin or hair 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.

Melanin in hair is an important factor that determines the hue of hair, and melanin produced in melanin-forming cells present in the hair sac is transmitted to keratinocytes of hair cortical cells and then moves upward as the hair grows. do. Hair bleaching, which is recognized as a phenomenon of physiological aging, is induced by a decrease in melanin production due to a decrease in the number of hair melanin-forming cells and a decrease in the function of melanin-forming cells. Currently, hair dyeing is used as a solution to whitening hair, but hair dyeing is a temporary method that needs to be re-dyed as the hair grows, and is included in the hair dye. The existing ingredients become irritating ingredients and cause contact dermatitis and skin allergies, which is a problem.

In addition, when an abnormality occurs in melanin biosynthesis, abnormal whitening of the skin appears due to abnormal coloring, which causes diseases such as bleaching (albinism) and vitiligo.

U.S. Patent Application Publication No. 2013-0296349 U.S. Patent Application Publication No. 2011-0015201

FIRESTONE, A. J. et al., Small-molecule Inhibitors of the AAA-ATPase Motor Cytoplasmie Dynein., Nature, 2012, vol. 484, no. 7392, pages 125-129 KIM, E.S. et al., Inhibition of Autophagy Suppresses Sertraline-mediated Primary Ciliogenesis in Rentinal Pigment Epithelium Cells., PLoS One, 2015, vol. 10, no. 2, thesis no. E011890 CHOI, H. et al., Primary Cilia Negatively Regulate Melanogenesis in Melanocytes and Pigmentation in a Human Skin Model., PLoS One, December 2016, vol. 11, no. 12, thesis no. E0168025

In one aspect, the present invention relates to 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile, an optical or stereoisomer thereof. Composition for blackening hair, preventing whitening of hair, preventing melanin-lowering disease, or improving melanin-lowering disease, which contains the acceptable salt, its hydrate, or its solvate as an active ingredient. The purpose is to provide products to develop the fields of white hair care and melanin-lowering diseases, and to improve the welfare of related consumers and patients.

In order to solve the above problems, the present invention presents on one aspect, 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile, For blackening of hair containing the optical or stereoisomer, its acceptable salt, its hydrate, or its solvate as an active ingredient, for preventing whitening of hair, for preventing melanin-lowering diseases, and for lowering melanin. A composition for treating a sexual disease or for improving a melanin-lowering disease is provided.

In another aspect, the present invention describes 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile, an optical or stereoisomer thereof. , A method for blackening hair, a method for preventing whitening of hair, a method for preventing melanin-lowering diseases, a method for preventing melanin-lowering diseases, including administration of an acceptable salt thereof, a hydrate thereof, or a solvate thereof to an individual. A method for improving melanin, or a method for treating a melanin-lowering disease.

In another aspect, the present invention is used for blackening hair, preventing whitening of hair, preventing melanin-lowering diseases, treating melanin-lowering diseases, or improving melanin-lowering diseases. 2,4-Dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazoliniriden) -β-oxo-benzenepropanenitrile, its optical or steric isomer, its acceptable salt, its water A Japanese product or a solvate thereof is provided.

In another aspect, the present invention comprises 2,4 as an active ingredient for blackening hair, preventing whitening of hair, preventing melanin-lowering diseases, treating melanin-lowering diseases, or improving melanin-lowering diseases. -Dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile, its optical or stereoisomer, its acceptable salt, its hydrate, Alternatively, the solvate thereof provides a non-therapeutic cosmetic use.

In other aspects, the present invention comprises a hair blackening composition, a hair whitening prevention composition, a melanin-lowering disease preventive composition, a melanin-lowering disease therapeutic composition, or a melanin-lowering disease. 2,4-Dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile, its optical or stereoscopic, for use in the production of the composition for improvement of The use of an isomer, an acceptable salt thereof, a hydrate thereof, or a solvate thereof is provided.

In another aspect of the invention, the composition may be a pharmaceutical composition or a cosmetic composition.

2,4-Dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile, its optical or stereoisomer, according to one aspect of the present invention. When a composition containing the acceptable salt, its hydrate, or its solvate as an active ingredient is used, blackening of hair, prevention of whitening of hair, prevention of melanin-lowering disease, or melanin-lowering disease It can be treated and effectively planned.

It is a figure which confirmed the effect of 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazoliniriden) -β-oxo-benzenepropanenitrile on the content of intracellular melanin. Western blot showed the effect of 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazoliniriden) -β-oxo-benzenepropanenitrile on the expression of tyrosinase and TRP1 protein. It is a figure. Effect of 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylidene) -β-oxo-benzenepropanenitrile on intracellular melanin content in B16F1 melanoma cell line It is a figure shown. Effect of 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile on tyrosinase and TRP1 protein expression in B16F1 melanoma cell line It is a figure confirmed by Western blotting.

Hereinafter, the present specification will be described in detail.

As used herein, the term "hair" refers to a filamentous structure composed of keratinized epithelial cells that appears on the surface of the skin, and specifically means hair of the body, which includes hair and body hair. It is a concept in a broad sense.

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.

As used herein, the term "blackening" means an increase in colored pigments in the skin or hair, and in particular, a phenomenon in which the amount of melanin pigments increases and the skin or hair becomes darker in color. Can be done. For example, blackening of hair includes a phenomenon in which hair turns black, and also includes prevention and prevention of whitening of hair and whitening of hair.

As used herein, "whitening" means a decrease in colored pigments in the skin or hair, and in particular, a phenomenon in which the amount of melanin pigments decreases and the skin or hair becomes lighter in color. obtain. For example, bleaching or whitening of hair includes the phenomenon that the hair turns white.

In the present specification, the "hair blackening effect substance" or the "hair whitening prevention effect substance" is a general term for substances that prevent the brightness of the hair from being darkened or the whitening of the hair from further progressing. It may be there. In addition, it may be a general term for substances that induce, improve, and promote the deposition of colored pigments, particularly melanin, such as single compounds, polymers, DNA, RNA, miRNA, peptides, and proteins. However, it is not limited to these.

As used herein, the term "melanin-lowering disease" includes a disease in which melanin deposition is abnormally suppressed due to abnormal synthesis or decomposition of melanin pigment. For example, vitiligo (albinism), leukoderma, partial vitiligo, vitiligo, vitiligo, quadricrome vitiligo, punctate vitiligo, idiopathic drops. Hypomelanosis, punctate vitiligo, syndromic Albinism (eg, Alezzandrinisyndrome, Hermansky-Pudlak syndrome, Hermansky-Pudlac syndrome, Chedia) (Chediak-Higashi syndrome), Griscily syndrome (Griscelli syndrome) (Elegarde syndrome), Griscily syndrome type 2 (Griscelli syndrome type3) and Griscily syndrome Wardenburg syndrome, Tiets syndrome, Cross-MuKusikk-Breen syndrome, Cross-MuKusick-Breen syndrome, ABCD syndrome, Vitiligo, Vitiligo, Vitiligo, Vitiligo, Vitiligo, Vitiligo deafness syndrome, Vogt-Koyanagi-Harada syndrome, ocular skin vitiligo (oculocutaneous albinism), hypomelanosis (hypomelanosis) ), Phylloid hypomelanosis, progressive patchy hypomelanosis), vitiligo patchy, mottled peebaldism, depigmented vitiligo, depigmented vitiligo (Post-inflammatory pigmentation, postinflammatory hyperpigmentation), white vitiligo (pityriasis alba), Vagabond's vitiligo (Vagabond's leukome) randorma), Yemenite deaf-blind hyperpigmentation syndrome, Wende-Buckus syndrome, Wolonoff ring (Woronoff's leucism) )including.

As used herein, the term "melanin-lowering disease preventive, ameliorating and therapeutically effective substance" refers to a substance that prevents or suppresses melanin-lowering disease in advance, a substance that reduces or alleviates the symptoms of melanin-lowering disease. It contains substances that eliminate the cause of melanin-lowering diseases, substances that promote melanin synthesis, substances that prevent suppression of melanin synthesis, and substances that suppress the decomposition of melanin, such as the entire skin, the entire hair, and specific hair. Or, it is a general term for substances that act to darken the brightness of specific skin areas or induce, improve, or enhance the deposition of melanin pigment, and is a single compound, polymer, DNA, RNA, etc. It may include, but is not limited to, peptides, proteins and the like.

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, "pharmaceutically acceptable" is 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 similar regulatory bodies, or listed in the Food Code, Health Function Food Standards, or General Pharmacopoeia, or otherwise in general It means that it is recognized as being described in a typical document.

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.

In one aspect, the present invention relates to 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile, an optical or stereoisomer thereof. Provided is a composition for darkening hair or preventing whitening of hair, which comprises an acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

In another aspect, the present invention describes 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile, its optical or stereoisomer. , A composition for preventing and treating melanin-lowering diseases, which comprises an acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

In the present specification, the 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile (2,4-Dichloro-a- (2,4-Dichloro-a-) 3,4-dihydro-4-oxo-2 (1H) -quinazalylidene) -β-oxo-benzenepropanitile) may be expressed as "substance A" and may contain a compound represented by the following chemical formula 1.

According to another aspect of the present invention, 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. May be good.

According to one embodiment, the 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile enhances the production or deposition of melanin. It may be something to make.

According to other embodiments, the 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile is tyrosinase or TRP1. It may enhance the expression of (Tyrosinase-Related Protein 1). Tyrosinase is an enzyme involved in the formation of melanin, and when the expression of tyrosinase is enhanced, the production or deposition of melanin is increased. In addition, TRP1 is a protein involved in the synthesis of melanin, and like tyrosinase, when the expression of TRP1 is enhanced, the production or deposition of melanin is increased.

After the treatment of the compound, the production or deposition of melanin is promoted, and the expression of tyrosinase and TRP1 is higher than that before the treatment of the compound. It can be seen that it can be used as an active ingredient of a substance. Further, it can be seen that when the compound is treated, the number of cells expressing tyrosinase or TRP1 increases, and through this, the compound can be used as an active ingredient of the composition. Melanin in hair is an important factor that determines the hue of hair, and melanin produced in melanin-forming cells present in the hair sac is transmitted to keratinocytes of hair cortical cells and then moves upward as the hair grows. Therefore, hair bleaching is induced by a decrease in the number of hair melanin-forming cells and a decrease in melanin production due to a decrease in the function of melanin-forming cells. The 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazoliniriden) -β-oxo-benzenepropanenitrile is used in melanin-forming cells such as melan-a cells and B16F1 cells. Since it enhances the expression of tyrosinase and TRP1 and increases the production and deposition of melanin, it can be used in compositions for blackening hair and preventing whitening of hair.

According to other embodiment, the melanin-lowering diseases include bleaching, vitiligo, depigmented mother's spot, pityriasis alba, albinism, post-inflammatory depigmentation, patchy sclerosis, partial vitiligo, idiopathic. It may be one or more selected from the group consisting of drip-like hypomelaninosis and punctate albinism. Melanin is observed from the outer skin hair, hair, skin, head, eye, etc. of animals, and when melanin production is insufficient, deposition is suppressed, or excessive decomposition occurs, melanin-lowering disease occurs. May get sick. In one aspect, the present invention enhances the production or deposition of melanin and enhances the expression of tyrosinase and TRP1, and thus can be used for the prevention, amelioration, and treatment of the melanin-lowering disease.

In one aspect, the present invention produces melanin in melanin-forming cells by 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile. Was confirmed to be enhanced, and it was confirmed that the expression of tyrosinase and TRP1 was enhanced. Using the efficacy of 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazoliniriden) -β-oxo-benzenepropanenitrile, the disease related to the decrease in melanin. It is possible to prevent, improve, and treat the symptoms and symptoms.

According to another aspect of the present invention, 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile in the composition. The content of the isomer, an acceptable salt thereof, a hydrate thereof, or a solvate thereof may be in the range of 0.0001% by mass to 20% by mass with respect to the total mass of the composition. 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 or more, based on the total mass of the composition. 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, It may be 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 mass 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 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile, its optics or steric. The dose of the isomer, 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. The doses are 0.0001 mg / kg / day or more, 0.0005 mg / kg / day or more, 0.001 mg / kg / day or more, 0.005 mg / kg / day or more, 0.01 mg / kg / day or more. Above, 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 / 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 / kg / day or more That may be the above. 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 embodiment of the present invention, the composition may be 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 osmotic pressure regulation, and other therapeutically useful. Substances may be further included and may be emulsified in the form of various oral or parenteral administrations by conventional methods.

Examples of the oral administration agent include tablets, pills, hard and soft capsules, liquids, suspensions, 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 another embodiment of the present invention, the composition may be a cosmetic composition for blackening hair, preventing whitening of hair, preventing or ameliorating melanin-lowering diseases.

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, bactericides, antioxidants, plant extracts, pH adjustments. Examples include agents, alcohols, pigments, fragrances, blood circulation promoters, cooling 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. Talk, 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 metahydroxydo, 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.

According to another aspect of the invention, the composition may be a food or health food composition.

The food or health food composition may be in a liquid or solid dosage form, and includes, for example, various foods, beverages, gums, teas, vitamin compounds, health functional foods, dietary supplements, and the like. , Powders, granules, tablets, capsules, or may be used in the form of beverages. In addition to the active ingredient, the food composition of each dosage form may be appropriately selected and blended by a person skilled in the art according to the dosage form or the purpose of use according to the dosage form or the purpose of use. A synergistic effect may be achieved when applied at the same time as the raw material.

The liquid component that can be contained in addition to the active ingredient disclosed in the present specification is not particularly limited, and various flavoring agents, natural carbohydrates, and the like may be contained as additional components as in ordinary beverages. Examples of the natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the flavoring agent, a natural flavoring agent (thaumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.)) and a synthetic flavoring agent (saccharin, aspartame, etc.) may be advantageously used. The proportion of said natural carbohydrates may be generally about 1-20 g per 100 ml of the compositions disclosed herein, and in one aspect may be about 5-12 g.

In one aspect, the food composition comprises a variety of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acids and the like. It may contain salts, alginic acids and salts thereof, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In another aspect, it may include pulp for the production of natural fruit juices and vegetable beverages. These components may be used independently or in combination. The proportion of the additive may vary, but is generally selected in the range of 0.001 to about 20 parts by mass per 100 parts by mass of the composition disclosed herein.

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.

Reagents 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile, α-MSH, arbutin, and kojic acid acid) was 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] Confirmation of melanin production and regulation of related enzymes and proteins 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile (Substance A) is a substance represented by the following chemical formula 1.

Melanocytes were treated with substance A (10 μM, 30 μM), respectively, and 48 hours later, the cell melanin content and the expression levels of tyrosinase and TRP1 protein were analyzed by Western blotting. As a result, serum starvation slightly reduced the content of melanin and the expression of tyrosinase and TRP1 protein in melan-a cells, but then the treatment with substance A resulted in an increase in melanin synthesis and expression of tyrosinase and TRP1 protein. Enhancement was confirmed (FIGS. 1 and 2, "substance A10" is 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropane. The group to which 10 μM of nitrile was administered is shown, and “substance A30” is 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile 30 μM. Indicates the group to which it was administered).

It was confirmed whether such a phenomenon also appears in the B16F1 mouse melanoma cell line. Specifically, B16F1 cells were treated with substance A (10 μM, 30 μM) in serum starvation medium and then cultured for 48 hours. Cellular melanin content was then measured (FIG. 3) and tyrosinase and TRP1 were analyzed by Western blot (FIG. 4).

As a result, the synthesis of melanin by the substance A and the expression of the melanin-producing enzyme were also induced in the B16F1 cell line (FIGS. 3 to 4).

Integrating these contents and inducing the synthesis of melanin and the expression of melanin-producing enzymes by substance A, it is possible to promote the production of melanin in the melanin-forming cells in the skin and hair sac, thereby causing hair. Since it is possible to prevent blackening and whitening of hair, one aspect of the present invention can be used for blackening of hair and whitening of hair. In addition, since it is possible to promote the production of melanin for a specific part of the skin or the entire skin by using such a feature of the present invention, the present invention can be used to prevent or treat melanin-lowering diseases and symptoms. And can be improved.

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 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile 20 mg, L-carnitine 80- 140 mg, 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 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile 20 mg, galactooligosaccharide 200 mg, lactose 60 mg and After mixing 140 mg of maltose and granulating it using a fluidized layer dryer, 6 mg of sugar ester was added and tableted with a tableting machine to produce tablets.

[Dosage Form Example 3] Granules 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile 20 mg, anhydrous crystalline glucose 250 mg and starch 550 mg was mixed, and the granules were formed into granules using a fluidized bed granulator, and then packed in a packet to produce granules.

[Formation Example 4] Drink 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile 20 mg, glucose 10 g, citric acid 0 After mixing .6 g and 25 g of liquid oligosaccharide, add 300 ml of purified water and fill each bottle 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 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile 50 mg, sterilized distillation for injection in an appropriate amount An injection was prepared according to a usual method using water and an appropriate amount of a pH adjuster.

[Dosage form example 6] Soft lotion (skin lotion)
2,4-Dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile 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 0.20% by mass, 1,3- A soft cosmetic solution (skin lotion) was produced using 3.00% by mass of butylene glycol and purified water as the remaining amount.

[Dosage Form Example 7] Cream 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile 3.00% by mass, polyethylene glycol mono Steerate 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 6.00% by mass, spingosaccharide A creamy preparation was produced using 1.00% by mass of lipid, 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 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile 3.00% by mass, polyvinyl alcohol 13 .00% by mass, L-ascorbic acid-2-maganoic acid salt 1.00% by mass, lauroyl hydroxyproline 1.00% by mass, water-soluble collagen (1% aqueous solution) 2.00% by mass, 1,3-butylene A pack was produced using 3.00% by mass of glycol, 5.00% by mass of ethanol, and purified water as 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. 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] Healthy Beverage

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 (10)

2,4-Dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile (2,4-Dichloro-a- (3,4-dihydro-) 4-oxo-2 (1H) -quinazadiolidene) -β-oxo-benzenepropanitile), its optical or steric isomer, its acceptable salt, its hydrate, or its solvate as an active ingredient in black hair Composition for chemistry or prevention of bleaching of hair. 2,4-Dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylidene) -β-oxo-benzenepropanenitrile, its optical or steric isomer, its acceptable salt, its water A composition for preventing and treating a melanin-lowering disease containing a Japanese product or a solvate thereof as an active ingredient. The 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylidene) -β-oxo-benzenepropanenitrile enhances the production or deposition of melanin, claim. Item 2. The composition according to Item 1 or 2. The 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylene) -β-oxo-benzenepropanenitrile is a tyrosinase or TRP1 (Tyrosinase-Related Protein 1). The composition according to claim 1 or 2, which enhances the expression of. 2,4-Dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinidene) -β-oxo-benzenepropanenitrile in the composition, an isomer thereof, an acceptable salt thereof, The composition according to claim 1 or 2, wherein the content of the hydrate or the solvate thereof is in the range of 0.0001% by mass to 20% by mass with respect to the total mass of the composition. The 2,4-dichloro-a- (3,4-dihydro-4-oxo-2 (1H) -quinazolinylidene) -β-oxo-benzenepropanenitrile, its optical or stereoisomer, its acceptable salt, and the like. The composition according to claim 1 or 2, wherein the dose of the hydrate or a solvate thereof is in the range of 0.0001 mg / kg / day to 20 mg / kg / day. The melanin-lowering diseases include bleaching, vitiligo, depigmented mother's spot, pityriasis alba, albinism, post-inflammatory depigmentation, patchy emphysema, partial leukoderma, idiopathic droplet hypomelaninosis and The composition according to claim 2, which is one or more selected from the group consisting of punctate albinism. The composition according to claim 1 or 2, wherein the composition is a pharmaceutical composition. The composition is a whitening prevention of粧料composition or hair darkening of the hair, the composition of claim 1. The composition according to claim 2, wherein the composition is a cosmetic composition for preventing and improving melanin-lowering diseases.

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