KR102672900B1 - COMPOSITION FOR MELANIN-ENHANCING COMPRISING 2,4-DICHLORO-A-(3,4-DIHYDRO-4-OXO-2(1H)-QUINAZOLINYLIDENE)-β-OXO-BENZENEPROPANENITRILE - Google Patents

Abstract

In one aspect, the present invention relates to 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, its optical or stereoscopic It relates to a composition for blackening hair, preventing hair whitening, and preventing and improving melanin-lowering diseases, comprising an isomer, an acceptable salt thereof, a hydrate, or a solvate thereof as an active ingredient, and through one aspect of the present invention, 2 ,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile can provide new uses and effects to consumers, It can promote the development of related industries.

Description

Composition for melanin enhancement comprising 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile {COMPOSITION FOR MELANIN- ENHANCING COMPRISING 2,4-DICHLORO-A-(3,4-DIHYDRO-4-OXO-2(1H)-QUINAZOLINYLIDENE)-β-OXO-BENZENEPROPANENITRILE}

The present specification discloses a hair blackening product containing 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile and melanin reduction. It relates to a composition for preventing sexual diseases or treating melanin-lowering diseases.

Melanin is a phenol-based biopolymer in the form of a complex of black pigment and protein. The browning that occurs on the cut surfaces of apples, potatoes, and bananas when exposed to the air, or the browning that occurs in animal skin, feathers, skin, hair, eyes, etc. is observed. When melanin is overproduced and deposited, spots, freckles, etc. are formed and are directly related to hair darkening. Melanin can also accelerate skin or hair aging and cause skin cancer.

Melanocyte stimulating hormone (MSH) is secreted by ultraviolet rays, inflammation, hormones, etc., and MSH reacts with receptors to increase cAMP in melanin-forming cells to synthesize melanin, and the synthesized melanin is absorbed into the melanin-forming cells. It is secreted externally and plays a role in protecting the skin or hair from ultraviolet rays. Melanin synthesis is mainly regulated by α-MSH, and proteins involved in melanin synthesis include MITF, TYR, TRP1, and TRP2.

Melanin in hair is an important factor in determining hair color. Melanin produced within melanin-forming cells present in hair follicles is delivered to hair cortical cells keratinocytes and then moves upward along with hair growth. Hair whitening, which is recognized as a phenomenon of physiological aging, is caused 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, dyeing is being used as a solution to hair bleaching, but dyeing is a temporary method and requires re-dying due to gray hair growth, and the ingredients contained in the dye of hair dye are irritating and can cause contact dermatitis or skin allergies. It is causing a problem.

In addition, if there is an abnormality in melanin biosynthesis, abnormal coloring causes abnormal whitening of the skin, which may lead to diseases such as albinism and vitiligo.

J. A. SWIFT, Transfer of Melanin Granules from Melanocytes to the Cortical Cells of Human Hair, Nature 203, 976-977 (29 August 1964)

In one aspect, the present invention provides 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, its optical or stereoscopic Provided is a composition for hair blackening, preventing hair whitening, preventing melanin-lowering diseases, or improving melanin-lowering diseases, comprising an isomer, an acceptable salt thereof, a hydrate or a solvate thereof as an active ingredient, and providing a composition for treating white hair. The purpose is to promote the development of care and low-melanin disease-related fields and to improve the welfare of related consumers and patients.

In order to solve the above problem, the present invention in one aspect, 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropane A composition containing nitrile, its optical or stereoisomer, its acceptable salt, its hydrate, or its solvate as an active ingredient for hair blackening, preventing hair whitening, preventing melanin-lowering diseases, or improving melanin-lowering diseases. provides.

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

According to one aspect of the present invention, 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, its optical or When using a composition containing a stereoisomer, an acceptable salt thereof, a hydrate, or a solvate thereof as an active ingredient, it can be used to prevent hair darkening, hair whitening, prevent melanin-lowering diseases, or treat or improve melanin-lowering diseases. It can be promoted effectively.

Figure 1 shows the effect of 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile on intracellular melanin content. It has been confirmed.
Figure 2 shows tyrosinase and TRP1 protein expression of 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile. The effect on is shown by Western blot.
Figure 3 shows that 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile induces intracellular melanin in the B16F1 melanoma cell line. This shows the effect on content.
Figure 4 shows 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile in B16F1 melanoma cell line. The effect on TRP1 protein expression was confirmed by Western blot.

Hereinafter, the present invention will be described in detail.

In this specification, “hair” refers to a thread-like structure made of keratinized epithelial cells that emerges from the surface of the skin, and specifically refers to body hair, and is the broadest concept including hair and body hair.

In this specification, “skin” refers to the tissue that covers the body surface of an animal, and is the broadest concept that includes not only the tissue that covers the body surface such as the face or body, but also the scalp and hair.

As used herein, “blackening” refers to an increase in the colored pigment of the skin or hair, and in particular, may refer to a phenomenon in which the amount of melanin pigment increases, causing the skin or hair to appear dark in color. For example, hair darkening includes the phenomenon of hair turning black, and also includes preventing and preventing hair bleaching or graying.

As used herein, “whitening” refers to a decrease in the colored pigment of the skin or hair, and in particular, may refer to a phenomenon in which the amount of melanin pigment is reduced, causing the skin or hair to appear pale in color. For example, hair efflorescence or graying involves the phenomenon of hair turning gray.

In the present specification, “hair blackening effective substance” or “hair bleaching prevention effective substance” may be a general term for substances that darken the brightness of hair or prevent further hair bleaching. In addition, it can be a general term for substances that induce, improve, and enhance the deposition of colored pigments, especially melanin, and may include single compounds, polymers, DNA, RNA, peptides, proteins, etc., but is not limited thereto.

As used herein, “melanin-lowering disease” includes a disease in which melanin deposition is abnormally suppressed due to abnormalities in the synthesis or decomposition of melanin pigment. For example, albinism, leukoderma, partial albinism, tinnitus, vitiligo, quadrichrome vitiligo, vitiligo ponctue, idiopathic erythrohypopigmentation. Leukoderma petechiae, syndromic albinism (e.g. Alezzandrinisyndrome, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome, Griselli Griscelli syndrome (Elejalde syndrome, Griscelli syndrome type 2 and Griscelli syndrome type 3), Waardenburg syndrome, Titze syndrome (Tietz syndrome), Cross-MuKusick-Breen syndrome, ABCD syndrome, Albinism-deafness syndrome and Vogt-Koyanagi-Harada syndrome -Koyanagi-Harada syndrome), oculocutaneous albinism, hypomelanosis (idiopathic guttate hypomelanosis, phylloid hypomelanosis, progressive patchy hypomelanosis ( progressive macular hypomelanosis), ecchymosis, piebaldism, nevus depigmentosus, postinflammatory hypopigmentation, pityriasis alba, vagabond vitiligo. (Vagabond's leukomelanoderma), Yemenite deaf-blind hypopigmentation syndrome, Wende-Bauckus syndrome, Woronoff's ring, melanism, and Lu Includes leucism.

In this specification, “substances effective in preventing, improving, and treating hypomelanin diseases” refer to substances that prevent or suppress the development of hypomelanin diseases in advance, substances that reduce or alleviate the symptoms of hypomelanin diseases, and substances that reduce melanin levels. It includes substances that relieve the cause of sexual diseases, substances that enhance melanin synthesis, substances that prevent the inhibition of melanin synthesis, and substances that suppress the decomposition of melanin, and are used to treat the entire skin, all hair, specific hair, or specific skin areas. It is a general term for substances that darken the brightness or induce, improve, and enhance the deposition of melanin pigment, and may include, but is not limited to, single compounds, polymers, DNA, RNA, peptides, proteins, etc.

As used herein, “isomer” specifically refers to optical isomers (e.g., essentially pure enantiomers, essentially pure stereoisomers or mixtures thereof, as well as essentially pure diastereomers). , stereoisomers, conformation isomers (i.e. isomers that differ only in the angle of one or more chemical bonds), structural isomers, position isomers (especially tautomers), or geometric isomers. (e.g., cis-trans isomers).

As used herein, "essentially pure", for example, when used in relation to enantiomers, stereoisomers, or diastereomers, includes about 90 specific compounds of which enantiomers, stereoisomers, or diastereomers are present. % or more, preferably at least about 95%, more preferably at least about 97% or at least about 98%, even more preferably at least about 99%, even more preferably at least about 99.5% (w/w) It means to exist.

As used herein, “acceptable” means that it can be or has been approved by a government or equivalent regulatory agency for use in animals, more specifically in humans, by avoiding significant toxic effects when used in conventional or medicinal dosages. It means that it is recognized as being received, listed in the Food Code, Codex of Health Functional Foods, or General Pharmacopoeia, or listed in other general literature.

As used herein, “acceptable salt” refers to a salt according to one aspect of the present invention that is conventionally or pharmaceutically acceptable and has the desired activity of the parent compound. The salts are (1) formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, 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 -acid addition salts formed with organic acids such as butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid; or (2) a salt formed when an acidic proton present in the parent compound is replaced.

In this specification, “hydrate” refers to a compound to which water is bound, and is a broad concept that includes embedded compounds in which there is no chemical bond between water and the compound.

In this specification, “solvate” refers to a higher-order compound formed between a solute molecule or ion and a solvent molecule or ion.

In one aspect, the present invention provides 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, its optical or stereoscopic Provided is a composition for blackening hair or preventing hair whitening, comprising an isomer, an acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

In another aspect, the present invention relates to 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, optical or stereoisomers thereof. , an acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient, and a composition for preventing and treating melanin-lowering diseases is provided.

In the present specification, the 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile (2,4-Dichloro-a -(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile) may be expressed as “Substance A” and may include a compound represented by Formula 1 below.

[Formula 1]

According to another aspect of the present invention, the compound may be obtained through synthesis, may be obtained by processing other materials, or may be derived from living organisms or microorganisms.

According to one embodiment, the 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile is used to produce or deposit melanin. It may be to promote .

According to another embodiment, the 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile is tyrosinase. Alternatively, it may enhance the expression of TRP1 (Tyrosinase-Related Protein 1). Tyrosinase is an enzyme involved in the formation of melanin, and when the expression of tyrosinase is increased, melanin production or deposition increases. Additionally, TRP1 is a protein involved in melanin synthesis, and like tyrosinase, when TRP1 expression is enhanced, melanin production or deposition increases.

After treatment with the compound, melanin production or deposition is enhanced, and the expression of tyrosinase and TRP1 is higher than before treatment with the compound. This shows that the compound can be used as an active ingredient in a composition for hair blackening or preventing hair whitening. In addition, when treated with the compound, the number of cells expressing tyrosinase or TRP1 increases, which shows that the compound can be used as an active ingredient in the composition. Melanin in hair is an important factor in determining hair color. Melanin produced within melanin-forming cells present in hair follicles is delivered to hair cortical cells keratinocytes and then moves upward along with hair growth. Hair bleaching is caused 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. The 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile is used in melanin-forming cells, melan-a cells and B16F1. Since it enhances the expression of tyrosinase and TRP1 in cells and increases melanin production and deposition, it can be used in compositions for hair blackening and preventing hair whitening.

According to another embodiment, the melanin-lowering disease is a group consisting of chlorosis, vitiligo, depigmented nevus, white pityriasis, pityriasis, post-inflammatory depigmentation, ecchymosis, partial leukoderma, idiopathic red hypopigmentation, and petechial leukoderma. There may be one or more selected from . Melanin is observed in the outer skin of animals, including feathers, fur, skin, hair, and eyes. If melanin is not produced properly, its deposition is suppressed, or its decomposition is excessive, melanin-lowering disease may result. In one aspect, the present invention enhances melanin production or deposition and the expression of tyrosinase and TRP1, so it can be used for the prevention, improvement, and treatment of the above-described melanin-lowering disease.

In one aspect, the present invention provides melanin forming cells by 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile. It was confirmed that melanin production was enhanced, and the expression of tyrosinase and TFP1 was confirmed to be enhanced. Using the efficacy of 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, the above related to melanin reduction It is possible to prevent, improve, and treat diseases or symptoms.

According to another aspect of the present invention, 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile in the composition, The content of its isomer, its acceptable salt, its hydrate, or its solvate may be 0.0001% by weight to 20% by weight based on the total weight of the composition. In addition, the content is 0.0001 weight% or more, 0.0005 weight% or more, 0.001 weight% or more, 0.005 weight% or more, 0.01 weight% or more, 0.05 weight% or more, 0.1 weight% or more, 0.3 weight% or more, 0.5 weight% or more based on the total weight of the composition. It may be at least 0.8% by weight, at least 1% by weight, at least 3% by weight, at least 5% by weight, at least 8% by weight, at least 10% by weight, at least 12% by weight, at least 15% by weight, or at least 18% by weight. there is. In addition, the content is 20% by weight or less, 18% by weight or less, 15% by weight or less, 12% by weight or less, 10% by weight or less, 8% by weight or less, 5% by weight or less, 3% by weight or less, 1 wt% or less, 0.8 wt% or less, 0.5 wt% or less, 0.3 wt% or less, 0.1 wt% or less, 0.05 wt% or less, 0.01 wt% or less, 0.005 wt% or less, 0.001 wt% or less, or 0.0005 wt% or less. You can.

According to another aspect of the present invention, the 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, its The dosage of the optical or stereoisomer, its acceptable salt, hydrate, or solvate thereof may be 0.0001 mg/kg/day to 20 mg/kg/day. In addition, the dosage is 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, 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. You can. In addition, the dosage is 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 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. You can.

According to one embodiment of the present invention, the composition may be a pharmaceutical composition.

The pharmaceutical composition may further contain pharmaceutical auxiliaries such as preservatives, preservatives, stabilizers, wetting agents or emulsification accelerators, salts and/or buffers for adjusting osmotic pressure, and other therapeutically useful substances, and may be used in a conventional manner. Accordingly, it can be formulated into various oral or parenteral dosage forms.

The oral administration agents include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, powders, powders, fine granules, granules, pellets, etc., and these formulations contain surfactants in addition to the active ingredients. , may contain diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (e.g. silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycol). . Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, and in some cases starch, agar, alginic acid or its sodium salt. It may contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavoring agents, and sweeteners. The tablets can be prepared by conventional mixing, granulating or coating methods.

In addition, the parenteral dosage form may be a transdermal dosage form, for example, injections, drops, ointments, lotions, gels, creams, sprays, suspensions, emulsions, suppositories, patches, etc. It may be, but is not limited to this.

The pharmaceutical composition may be administered parenterally, rectally, topically, transdermally, subcutaneously, etc.

Determination of the dosage of the active ingredient is within the level of a person skilled in the art, and the daily dosage of the drug may vary depending on various factors such as the degree of progression of the subject to be administered, time of onset, age, health condition, and complications.

The pharmaceutical composition may be an external preparation for the skin, and the external preparation for the skin is a general term that can include anything applied outside the skin, and may include various dosage forms of drugs or quasi-drugs.

According to another embodiment of the present invention, the composition may be a cosmetic composition for blackening hair, preventing hair whitening, or preventing or improving melanin-lowering diseases.

The cosmetic composition may additionally include functional additives and ingredients included in general cosmetic compositions. The functional additive may include ingredients selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extract. Other ingredients included include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohol, pigments, fragrances, and blood circulation agents. Examples include accelerators, cooling agents, restrictors, and purified water.

The formulation of the cosmetic composition is not particularly limited and can be appropriately selected depending on the purpose. For example, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, foundation, essence, nutritional essence, pack, soap, cleansing. It may be manufactured in one or more formulations selected from the group consisting of foam, cleansing lotion, cleansing cream, body lotion, and body cleanser, but is not limited thereto.

When the formulation according to one aspect of the present invention is a paste, cream, or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide are used as carrier ingredients. etc. can be used.

When the formulation according to one aspect of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the formulation is a spray, chlorofluoro May contain propellants such as hydrocarbons, propane/butane or dimethyl ether.

When the formulation according to one aspect of the present invention is a solution or emulsion, a solvent, solvating agent, or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, These include propylene glycol, 1,3-butyl glycol oil, glycerol fatty esters, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation according to one aspect of the present invention is a suspension, the carrier component includes water, a liquid diluent such as ethanol or propylene glycol, and a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester. , microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant, etc. can be used.

When the formulation according to one aspect of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, and sarcosylate. Nate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linoline derivative, or ethoxylated glycerol fatty acid ester can be used.

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

The food or health food composition may be in a liquid or solid form, and includes, for example, various foods, beverages, gums, teas, vitamin complexes, health functional foods, health supplements, powders, granules, tablets, etc. It can be used in capsule or beverage form. In addition to the active ingredients, the food composition of each formulation can be appropriately selected and mixed by a person skilled in the art according to the formulation or purpose of use, without difficulty, and a synergistic effect can occur when applied simultaneously with other raw materials.

There are no particular restrictions on the liquid ingredients that can be contained in addition to the active ingredients disclosed in this specification, and, like regular beverages, various flavoring agents or natural carbohydrates can be included as additional ingredients. Examples of the natural carbohydrates include monosaccharides, disaccharides such as glucose and fructose, polysaccharides such as maltose and sucrose, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. etc. As the above flavoring agents, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrate may be generally about 1 to 20 g, in one aspect, about 5 to 12 g per 100 ml of the composition disclosed herein.

In one aspect, the food composition includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts. , organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In another aspect, it may include pulp for the production of natural fruit juices and vegetable drinks. The above ingredients can be used independently or in combination. The ratio of the additive may vary, but is generally selected in the range of 0.001 to about 20 parts by weight per 100 parts by weight of the composition disclosed herein.

Hereinafter, the configuration and effect of one aspect of the present invention will be described in more detail through examples and experimental examples. However, the examples below are provided only for illustrative purposes to aid understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

[Example 1]

Cell and skin model preparation

Normal human epidermal melanocytes (NHEMs, Cascade Biologics, Portland, OR, USA) were obtained, and Melan-A, a melanogenic cell derived from C57BL/6 J (black, a/a) mouse, was obtained. -A) Cell lines were 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).

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

reagent

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, α-MSH, arbutin, and co Kojic acid was obtained from Sigma-Aldrich (St. Louis, MO, USA).

melanin essay

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

western Blot analyze

All lysates were stored in 2 × Laemmli sample buffer, 62.5 mM Tris-HCl, pH 6.8, 25% [v/v] glycerol, 2% [w/v] SDS, 5% [v/v]. ] It was prepared using β-mercaptoethanol and 0.01% [w/v] bromophenol blue) (Bio-Rad, Hercules, CA, USA). All cell proteins were measured using Bradford solution (Bio-Rad). Afterwards, the sample was separated by SDS-PAGE and transferred to a 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 was incubated with specific primary antibodies. Cultured overnight.

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 (1:1000) were obtained from VJ Hearing (NIH, Bethesda, MD). For protein detection, the membrane was incubated with HRP-conjugated secondary antibody, and the signal was measured using SuperSignal ^® West Dura HRP Detection Kit (Pierce, Rockford, IL, USA).

statistical analysis

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

[Experimental Example 1] Confirmation of melanin production, related enzymes and protein regulation

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile (Substance A) is a substance represented by the following formula (1): .

[Formula 1]

Melan-a cells were treated with substance A (10 μM, 30 μM), respectively, and 48 hours later, cell melanin content and expression levels of tyrosinase and TRP1 proteins were analyzed by Western blot. As a result, serum starvation slightly decreased the melanin content and tyrosinase and TRP1 protein expression in Melan-a cells, but subsequent treatment with substance A increased melanin synthesis and enhanced tyrosinase and TRP1 protein expression ( 1 and 2, “Substance A 10” is 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile It represents the group administered 10 μM, and “Substance A 30” is 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo- (Represents the group administered 30 μM benzene propanenitrile).

It was confirmed whether this phenomenon also occurs in the B16F1 mouse melanoma cell line. Specifically, B16F1 cells were cultured for 48 hours after treatment with substance A (10 μM, 30 μM) in serum starvation medium. Afterwards, cell melanin content was measured (Figure 3), and tyrosinase and TRP1 were analyzed by Western blot (Figure 4).

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

Taking these details together, if melanin synthesis and the expression of melanin-producing enzymes are induced through substance A, melanin production can be enhanced in melanin-forming cells in the skin and hair follicles, thereby preventing blackening and whitening of hair. Therefore, the present invention can be used for hair blackening and white hair care. In addition, since this feature of the present invention can be used to enhance melanin production in a specific area of the skin or the entire skin, the present invention can be used to prevent, treat, and improve melanin-lowering diseases or symptoms. .

A formulation example of the composition according to one aspect of the present invention is described below, but it can also be applied in various other formulations, and this is not intended to limit the present invention, but is only intended to be described in detail.

[Formulation Example 1] Soft capsule formulation

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile 20mg, L-carnitine 80~140mg, soybean oil 180mg, 2 mg of palm oil, 8 mg of hydrogenated vegetable oil, 4 mg of yellow lead, and 6 mg of lecithin were mixed and filled into one capsule according to a conventional method to prepare a soft capsule.

[Formulation Example 2] Tablets

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile 20 mg, galactooligosaccharide 200 mg, lactose 60 mg and maltose 140 mg was mixed and granulated using a fluidized bed dryer, then 6 mg of sugar ester was added and tableted using a tablet press to prepare tablets.

[Formulation Example 3] Granules

Mix 20 mg of 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, 250 mg of anhydrous crystalline glucose, and 550 mg of starch. , granules were manufactured by forming them into granules using a fluidized bed granulator and then filling them into bags.

[Formulation example 4] Drink formulation

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile 20 mg, glucose 10 g, citric acid 0.6 g, and liquid oligosaccharide After mixing 25g, add 300ml of purified water and fill each bottle with 200ml. After filling the bottle, it was sterilized at 130°C for 4 to 5 seconds to prepare a drink.

[Formulation Example 5] Injectable

50 mg of 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, appropriate amount of sterile distilled water for injection, and appropriate amount of pH adjuster. An injection was prepared using a conventional method.

[Formulation Example 6] Soft lotion (skin lotion)

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile 3.0% by weight, L-ascorbic acid-2- Use 1.00% by weight of magnesium phosphate, 1.00% by weight of water-soluble collagen (1% aqueous solution), 0.10% by weight of sodium citrate, 0.05% by weight of citric acid, 0.20% by weight of licorice extract, 3.00% by weight of 1,3-butylene glycol, and the remaining amount of purified water. Thus, soft lotion (skin lotion) was manufactured.

[Formulation Example 7] Cream

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile 3.00% by weight, polyethylene glycol monostearate 2.00% by weight , self-emulsifying glycerin monostearate 5.00% by weight, propylene glycol 4.00% by weight, squalene 6.00% by weight, tri2-ethylhexaneglyceryl 6.00% by weight, sphingoglycolipid 1.00% by weight, 1,3-butylene glycol 7.00% by weight. A cream-like formulation was prepared using %, 5.00% by weight of beeswax, and the remaining amount of purified water.

[Formulation Example 8] Pack

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile 3.00% by weight, polyvinyl alcohol 13.00% by weight, L -Ascorbic acid-2-phosphate magnesium salt 1.00% by weight, lauroyl hydroxyproline 1.00% by weight, water-soluble collagen (1% aqueous solution) 2.00% by weight, 1,3-butylene glycol 3.00% by weight, ethanol 5.00% by weight , a pack was prepared using the remaining amount of purified water.

[Formulation Example 9] Health food

Health food was prepared by a conventional method according to the composition shown in the table below.

ingredient content 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile 10mg vitamin mixture Vitamin A Acetate 70 ㎍ Vitamin E 1.0 mg Vitamin B1 0.13 mg Vitamin B2 0.15 mg Vitamin B6 0.5 mg Vitamin B12 0.2 μg Vitamin C 10 mg biotin 10 μg Nicotinic acid amide 1.7 mg folic acid 50 μg Calcium Pantothenate 0.5 mg mineral mixture Ferrous sulfate 1.75 mg zinc oxide 0.82 mg Magnesium Carbonate 25.3 mg Potassium Phosphate Monobasic 15 mg Dibasic Calcium Phosphate 55 mg Potassium citrate 90 mg calcium carbonate 100 mg Magnesium Chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture was prepared by mixing ingredients that are relatively suitable for health food as an example, but the mixing ratio may be modified arbitrarily. After mixing the above ingredients according to a normal health food manufacturing method, use the usual method. It can be used to manufacture health food compositions.

[Formulation example 10] Health drink

ingredient content 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile 15mg citric acid 1000 mg oligosaccharide 100 g plum concentrate 2g taurine 1g Purified water remaining amount total volume 900 ml

As shown in Table 2, the remaining amount of purified water was added to make a total volume of 900 ml, the ingredients were mixed according to a typical health drink manufacturing method, and then stirred and heated at 85°C for about 1 hour, and then the resulting solution was filtered. A health drink was prepared by obtaining it in a sterilized 2-liter container, sealing and sterilizing it, and then refrigerating it.

As above, specific parts of the present invention have been described in detail, and those skilled in the art will understand that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (9)

2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile (2,4-Dichloro-a-(3, For hair blackening containing 4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile), its optical or stereoisomer, its acceptable salt, its hydrate, or its solvate as an active ingredient. Or a composition for preventing hair bleaching. A composition for preventing and treating melanin-lowering diseases,
2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, optical or stereoisomers thereof, acceptable salts thereof, Contains its hydrate or its solvate as an active ingredient,
The melanin-lowering disease is one or more selected from the group consisting of chlorosis, vitiligo, depigmented nevus, white pityriasis, pityriasis, post-inflammatory depigmentation, ecchymosis, partial leukoderma, idiopathic red hypopigmentation, and petechial leukoderma. Composition for preventing and treating degenerative diseases. The method of claim 1 or 2, wherein the 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile is A composition that promotes melanin production or deposition. The method of claim 1 or 2, wherein the 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile is A composition that enhances the expression of tyrosinase or TRP1 (Tyrosinase-Related Protein 1). The method of claim 1 or 2, wherein 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropane in the composition A composition wherein the content of nitrile, its isomer, its acceptable salt, its hydrate, or its solvate is 0.0001% to 20% by weight based on the total weight of the composition. The method of claim 1 or 2, wherein 2,4-dichloro-a-(3,4-dihydro-4-oxo-2(1H)-quinazolinylidene)-β-oxo-benzenepropanenitrile, A composition wherein the dosage of its optical or stereoisomer, its acceptable salt, hydrate, or solvate thereof is from 0.0001 mg/kg/day to 20 mg/kg/day. delete The composition according to claim 1 or 2, wherein the composition is a pharmaceutical composition. The composition according to claim 1, wherein the composition is a cosmetic composition.