Composition for skin whitening and method for preparing same

Abstract

The present invention relates to a composition for skin whitening and a method for preparing same and, to a composition for skin whitening and a method for preparing same, in which a compound having a specific molecular structure and derived from a culture of Streptomyces sp. strains is used to inhibit the melanin production of melanoma cells and improve skin whitening effects.

Description

Composition for skin whitening and method for preparing the same

[0001] The present invention relates to a novel compound of chemical formula 1 and its use, comprising a compound having a specific molecular structure and a compound derived from a culture of a Streptomyces genus strain. This compound can exhibit excellent effects as a skin whitening composition that inhibits melanin production in melanoma cells and enhances skin whitening effects.

[0002] A variety of factors determine human skin color and contribute to the formation of freckles and blemishes. These include the activity of melanocytes (which produce melanin), blood vessel distribution, skin thickness, carotenoids, and bilirubin. The most crucial factor is melanin, which is produced by the action of various enzymes in melanocytes within the body. The formation of melanin is known to be influenced by a variety of physiological and environmental factors, including genetics, hormone secretion, stress, and ultraviolet rays.

[0003] Melanin, which determines human skin color, is produced in melanocytes. These melanocytes contain enzymes such as tyrosinase, which work together to form melanin, a dark-brown pigment, by polymerizing and oxidizing tyrosine, an amino acid that always exists in the body. This melanin then travels through the melanocyte's dendrites to epidermal cells called keratinocytes. Here, melanin plays a crucial role, forming a cap-like structure around the nucleus to protect genes from UV rays and scavenge free radicals to protect intracellular proteins. There are no enzymes in the body that break down melanin. However, when keratinocytes age, cease their function, and fall off from the epidermis, they are removed from the skin along with the keratinocytes. However, if melanin is produced in excess, it can cause hyperpigmentation, such as freckles, spots, and age spots. Accordingly, research has been conducted on whitening agents to suppress pigmentation in the skin by blocking excessive production of melanin, and inhibitors that inhibit tyrosinase activity, such as kojic acid and arbutin, hydroquinone, vitamin A, and representative antioxidants such as vitamin C and their derivatives have been reported.

[0004] However, existing whitening substances such as the above are often weak in efficacy or accompanied by cytotoxicity, so efforts are continuing to discover natural products that are safe and have high whitening efficacy.

[0005] The technical problem to be achieved by the present invention is to provide a skin whitening composition and a method for preparing the same, which has low toxicity and exhibits an excellent whitening effect, and can be used as a pharmaceutical composition for preventing or treating skin pigmentation or a cosmetic composition for preventing, improving, or alleviating skin pigmentation, using 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid {2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid} derived from a microorganism isolated from Jeju Island moss root marine sediment as an active ingredient, and which can be used as a pharmaceutical composition for preventing or treating skin pigmentation or a cosmetic composition for preventing, improving, or alleviating skin pigmentation.

[0006] However, the problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0007] One object of the present invention is to provide a compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof.

[0008] Another object of the present invention is to provide a cosmetic composition for skin whitening comprising a compound represented by the following chemical formula 1; or a cosmetically acceptable salt thereof.

[0009] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating skin pigmentation disease, comprising a compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof.

[0010] Another object of the present invention is to provide a food composition for preventing or improving skin pigmentation disease, comprising a compound represented by the following chemical formula 1; or a food-wise acceptable salt thereof.

[0011] Another object of the present invention is to provide a method for producing a cosmetic composition for skin whitening, comprising a compound represented by the following chemical formula 1; or a cosmetically acceptable salt thereof.

[0012] In this specification, when a part is said to "include" a certain component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise specifically stated.

[0013] In this specification, “A and / or B” means “A and B, or A or B.”

[0014] In this specification, “2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid” may mean a compound represented by the above chemical formula 1.

[0015] In this specification, “skin whitening” may mean preventing or improving blemishes, freckles, dark spots, and blemishes, and may mean including all activities that prevent, improve, or treat pigmentation on the skin.

[0016] The drawings attached to this specification illustrate preferred embodiments of the invention and, together with the description of the invention, serve to explain the principles of the invention, but are not intended to limit the scope of the invention. Furthermore, the shape, size, scale, or proportion of elements in the drawings included in this specification may be exaggerated for clarity.

[0017] Furthermore, to avoid confusion due to overlapping content, the description of overlapping content has been omitted. In other words, the content of the invention is not limited to the content described below, and the content of the invention should be interpreted based on the overall content of the invention.

[0018] Hereinafter, the present invention will be described in more detail.

[0019] The present inventors have made great efforts to discover a substance that exhibits an effective melanin synthesis inhibitory effect, and as a result, they have produced 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid from a marine actinomycete (Marine Streptomyces, APA-053) isolated from moss root marine sediment, and have discovered that this 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid can effectively inhibit melanin synthesis, thereby completing the present invention.

[0020] The present invention provides a compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof.

[0021] [Chemical Formula 1]

[0022]

[0023] The compound according to the present invention is 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid {2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid}.

[0024] In the present invention, “pharmaceutically acceptable salt” means a salt commonly used in the pharmaceutical industry, and includes, for example, inorganic ionic salts manufactured with calcium, potassium, sodium, and magnesium; inorganic acid salts manufactured with hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid, and sulfuric acid; organic acid salts manufactured with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, and hydroiodic acid; sulfonic acid salts manufactured with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid; Amino acid salts made from glycine, arginine, lysine, etc.; and amine salts made from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.; however, the types of salts meant in the present invention are not limited to these listed salts. Such pharmaceutically acceptable salts may also be acceptable salts in cosmetic or food applications.

[0025] In addition, the present invention is a skin whitening composition comprising a compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof.

[0026] More specifically, it may provide a cosmetic composition for skin whitening, a pharmaceutical composition for preventing or treating a skin pigmentation disease, or a food composition for preventing or improving a skin pigmentation disease.

[0027] According to one embodiment of the present invention, the compound represented by the chemical formula 1 may be included in one or more forms selected from the group consisting of the compound represented by the chemical formula 1; or a cosmetically, pharmaceutically, or food-wise acceptable salt, hydrate, solvate, isomer, or derivative thereof; and combinations thereof.

[0028] The present invention provides a cosmetic composition for skin whitening comprising a compound represented by the following chemical formula 1; or a cosmetically acceptable salt thereof.

[0029] [Chemical Formula 1]

[0030]

[0031] According to one embodiment of the present invention, the compound represented by the chemical formula 1 may be derived from a culture of a strain of the genus Streptomyces. It may also be produced by chemical synthesis.

[0032] According to one embodiment of the present invention, the compound represented by the chemical formula 1 may be derived from Streptomyces sp. APA-053 strain (Accession number: KCTC15538BP).

[0033] According to one embodiment of the present invention, the compound represented by the chemical formula 1 may be an extract or fraction of Streptomyces sp. APA-053 strain.

[0034] According to one embodiment of the present invention, the compound represented by the chemical formula 1 may inhibit melanin production of melanoma cells.

[0035] According to one embodiment of the present invention, the cosmetic composition for skin whitening may have a formulation selected from the group consisting of a solution, an ointment, an external ointment, a cream, an essence, a foam, a toner, a nourishing toner, an emollient, an emollient toner, a pack, a milky lotion, a makeup base, a soap, a cleanser, a bathing agent, a sunscreen, a sun oil, a suspension, an emulsion, a paste, a gel, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a foundation, a powder foundation, an emulsion foundation, a wax foundation, a patch, and a spray.

[0036] A skin whitening cosmetic composition according to one embodiment of the present invention is a safe substance because it is derived from a natural product, and can effectively inhibit melanin production, thereby preventing, treating, or improving skin pigmentation, and thus can be usefully used in the manufacture of a composition for developing a whitening pharmaceutical preparation or cosmetic.

[0037] According to one embodiment of the present invention, the cosmetic composition for skin whitening may include a salt of the compound represented by the above chemical formula 1, particularly in the form of a cosmetically acceptable salt. As the salt, any salt commonly used in the art may be used without limitation.

[0038] According to one embodiment of the present invention, the cosmetic composition for skin whitening may contain the compound represented by the chemical formula 1 or a salt thereof in an amount of 0.0025 wt% or more and 0.0100 wt% or less based on the total weight. Specifically, the amount may be adjusted depending on the form in which the cosmetic composition for skin whitening is manufactured, the method by which the compound is applied to the skin, the time it remains on the skin, etc., and when the cosmetic composition for skin whitening is manufactured as a pharmaceutical for dermatological treatment of hyperpigmentation of the skin, the composition may contain the compound in a higher concentration than when the composition is manufactured as a cosmetic for daily application to the skin.

[0039] According to one embodiment of the present invention, the compound may be derived from a culture of a strain of the genus Streptomyces. Specifically, since the genus Streptomyces APA-053 strain has the ability to produce the compound represented by the chemical formula 1, the culture of the genus Streptomyces APA-053 strain may be isolated and / or purified to obtain the compound represented by the chemical formula 1.

[0040] According to one embodiment of the present invention, the Streptomyces genus strain may be derived from Streptomyces sp. APA-053 strain (Accession No.: KCTC15538BP). Specifically, the Streptomyces sp. APA-053 strain (Accession No.: KCTC15538BP) corresponds to a strain capable of producing 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid, and corresponds to a strain that was confirmed by the inventors of the present invention to have the ability to produce 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid for the first time.

[0041] As a result of analyzing the strain according to the present invention through the 16S rRNA base sequence, it was possible to identify a strain with new genetic properties having the 16S rRNA base sequence of sequence number 6, and the strain was named Streptomyces sp. APA-053 strain, and was deposited with the Korea Collection for Type Cultures (KCTC, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 181 Ipsin-gil, Jeongeup-si, Jeollabuk-do, on August 4, 2023, and was assigned the designation KCTC15538BP.

[0042] The strain selected in the present invention can be used to produce a compound represented by chemical formula 1 included in the above skin whitening cosmetic composition.

[0043] According to one embodiment of the present invention, the compound represented by Chemical Formula 1 may be an extract or fraction of Streptomyces sp. APA-053 strain. Specifically, since the Streptomyces sp. APA-053 strain has the ability to produce the compound represented by Chemical Formula 1, the extract and / or fraction of the Streptomyces sp. APA-053 strain may be separated and / or purified to obtain the compound represented by Chemical Formula 1.

[0044] According to one embodiment of the present invention, when the compound represented by the above chemical formula 1 is an extract or fraction of Streptomyces sp. APA-053 strain, it may be included in an amount of 0.0010 wt% or more and 50 wt% or less based on the total weight. Specifically, when the compound represented by the above chemical formula 1 is an extract or fraction of Streptomyces sp. APA-053 strain, it may be included in an amount of 0.0100 wt% or more and 30 wt% or less, 0.1000 wt% or more and 10 wt% or less, or 1 wt% or more and 5 wt% or less based on the total weight. However, the content may be adjusted depending on the application area (face, neck, etc.) of the formulation of the present invention or the cosmetic composition for skin whitening, and it should not be understood that the scope of the present invention is limited by the content range.

[0045] According to one embodiment of the present invention, the skin whitening cosmetic composition may inhibit melanin production. Specifically, the skin whitening cosmetic composition may inhibit melanin production in melanoma cells. As described above, the skin whitening cosmetic composition may exhibit excellent skin whitening efficacy by inhibiting melanin production.

[0046] In the present specification, “inhibiting melanin production” may mean all mechanisms that lower the content of melanin in the skin and inhibit pigment deposition, such as inhibiting melanin synthesis in the skin and promoting the decomposition of produced melanin.

[0047] Specifically, the compound represented by the above chemical formula 1 exhibited an inhibitory activity that was approximately 37% higher than that of arbutin, and was confirmed to exhibit a very high melanin production inhibitory activity of approximately 45%, particularly at low concentrations. Consequently, a skin whitening cosmetic composition containing the above compound exhibits excellent melanin production inhibitory activity and can be usefully used in whitening preparations or cosmetics.

[0048] According to one embodiment of the present invention, the cosmetic composition for skin whitening may further include a cosmetically acceptable carrier or additive in addition to the compound represented by the chemical formula 1.

[0049] According to one embodiment of the present invention, the additive includes, but is not limited to, a fatty substance, an organic solvent, a solubilizer, a thickener, a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, an emulsifier, a filler, a metal ion sequestering agent, a chelating agent, a preservative, a vitamin, a blocking agent, a humectant, an oil, a dye, a pigment, a hydrophilic or lipophilic active agent, a lipid vesicle, and the like, and may include any known substance applicable to a cosmetic composition.

[0050] According to one embodiment of the present invention, the cosmetic composition may be formed into one or more formulations selected from the group consisting of paste, gel, emollient toner, nourishing toner, astringent toner, skin, lotion, essence, cream, massage cream, pack, makeup base, BB cream, foundation, powder, cleansing foam, cleansing cream, and cleansing water, but is not limited thereto, and may be formed into all known formulations.

[0051] According to one embodiment of the present invention, when the cosmetic composition is formed in a paste, cream or gel state, it may further include animal oil, vegetable oil, wax, paraffin, starch, tragacanth gum, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc.

[0052] According to one embodiment of the present invention, when the cosmetic composition is formed in the form of a solution or emulsion, it may further include a solvent, a solubilizer, an emulsifier, etc.

[0053] According to one embodiment of the present invention, the solvent, solubilizer, and emulsifier may include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, and the like.

[0054] According to one embodiment of the present invention, when the cosmetic composition is formed in the form of a suspension, it may further include a liquid diluent such as water, ethanol, or propylene glycol; a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester; microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or gum tragacanth.

[0055] According to one embodiment of the present invention, when the cosmetic composition is a cleansing foam, cleansing cream or cleansing water, it may further include fatty alcohol sulfate, fatty alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate-based compound, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, ethoxylated glycerol fatty acid ester, etc.

[0056] Additionally, for each type of cosmetic composition, appropriate ingredients can be selected and blended according to the cosmetic formulation or intended use. Since the blending ingredients and method can be achieved using conventional techniques, a detailed description thereof is omitted herein.

[0057] Additionally, according to one embodiment of the present invention, the skin whitening composition may be a pharmaceutical composition.

[0058] The present invention provides a pharmaceutical composition for preventing or treating skin pigmentation disease, comprising a compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof.

[0059] [Chemical Formula 1]

[0060]

[0061] In the present specification, “pharmaceutically acceptable salt” may mean any organic or inorganic addition salt of the compound represented by the above chemical formula 1, which has an effective effect at a concentration that is relatively non-toxic and harmless to the patient, and the side effects caused by the salt do not reduce the beneficial effects of the compound represented by the above chemical formula 1.

[0062] In the present invention, the skin pigmentation disease may be at least one selected from the group consisting of, for example, freckles, melasma, hyperpigmentation after UV exposure, hyperpigmentation after inflammation, lentigines, brown spots, and lentigines. That is, it is possible to prevent, improve, alleviate, or treat skin pigmentation and diseases related thereto, such as freckles, melasma, hyperpigmentation after UV exposure, hyperpigmentation after inflammation, lentigines, brown spots, or lentigines.

[0063] According to one embodiment of the present invention, the pharmaceutical composition can be used in a combined form with another pharmaceutically active compound.

[0064] According to one embodiment of the present invention, the pharmaceutical composition may be in the form of a known oral or parenteral administration preparation.

[0065] According to one embodiment of the present invention, the pharmaceutical composition may further include a diluent, excipient and / or carrier such as a filler, a bulking agent, a binder, a wetting agent, a disintegrating agent, a surfactant, etc.

[0066] According to one embodiment of the present invention, when the pharmaceutical composition is a solid preparation for oral administration, it may further include tablets, pills, powders, granules, capsules, etc., and such solid preparations may be prepared by mixing one or more compounds with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin.

[0067] According to one embodiment of the present invention, if the pharmaceutical composition is a preparation for parenteral administration, it may further include a sterile aqueous solution, non-aqueous solvent, suspension, emulsion, lyophilized preparation, or suppository.

[0068] According to one embodiment of the present invention, the non-aqueous solvent and suspending agent may be propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, etc.

[0069] According to one embodiment of the present invention, the base material of the suppository may be witepsol, macrogol, tween 61, cocoa butter, laurin butter, glycerogelatin, etc.

[0070] According to one embodiment of the present invention, the pharmaceutical composition can be administered to mammals such as rats, mice, livestock, and humans through various routes.

[0071] According to one embodiment of the present invention, the pharmaceutical composition may be administered by oral or parenteral administration (intraperitoneal, rectal, intravenous, intramuscular or subcutaneous injection, or transdermal administration), and the parenteral route is preferably transdermal administration, and among them, topical application may be most preferable.

[0072] According to one embodiment of the present invention, the dosage of the pharmaceutical composition may be adjusted according to the patient's condition and weight, the degree of disease, the drug form, the administration route and period, and specifically, may be administered once or several times a day in an amount of 0.1 to 100 mg / kg, but is not limited thereto.

[0073] According to one embodiment of the present invention, the parenteral route administration amount of the pharmaceutical composition varies depending on the patient's condition and weight, the degree of disease, the drug form, the route of administration, and the period of administration, but in the case of an external preparation, 1.0 to 3.0 ml may be applied once or several times per day, but is not limited thereto.

[0074] Additionally, according to one embodiment of the present invention, the skin whitening composition may be a food composition.

[0075] The present invention provides a food composition for preventing or improving skin pigmentation disease, comprising a compound represented by the following chemical formula 1; or a food-wise acceptable salt thereof.

[0076] [Chemical Formula 1]

[0077]

[0078] When the food composition of the present invention is used as a food additive, the food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. Generally, when manufacturing a food or beverage, the food composition of the present invention may be added in an amount of 15% by weight or less, preferably 10% by weight or less, relative to the raw materials.

[0079] There are no specific restrictions on the types of the above foods. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and all foods in the conventional sense are included.

[0080] The above beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. The above-mentioned natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, or synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrates may be appropriately determined by those skilled in the art.

[0081] In addition to the above, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the food composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in combination. The proportions of these additives may also be appropriately selected by those skilled in the art.

[0082] The food composition of the present invention can also be used as a health functional food. A health functional food is a food that emphasizes the bioregulatory function of food, and is a food that has added value by using physical, biochemical, and biotechnological methods to act and express for a specific purpose. The food composition of the present invention can be used as a health functional food. The ingredients of such a health functional food are designed and processed to sufficiently exert the body's body regulatory functions related to biodefense, regulation of body rhythm, and prevention and recovery from disease. The food composition may contain food-acceptable food additives, sweeteners, or functional raw materials. When the strain of the present invention is used as a health functional food (or health functional beverage additive), the novel strain may be added as is or used in combination with other foods or food ingredients, and may be used appropriately according to conventional methods. The mixing amount of the strains may be appropriately determined depending on the intended use (prevention, health or improvement, therapeutic treatment).

[0083] One embodiment of the present invention relates to a composition for producing a compound represented by the above chemical formula 1, comprising at least one selected from the group consisting of a culture comprising a strain selected from the group consisting of Streptomyces sp. APA-053 strain (Accession No.: KCTC15538BP), a dried product of the culture, a concentrate of the culture, and a combination thereof.

[0084] A composition for producing a compound represented by the chemical formula 1 according to one embodiment of the present invention can increase the yield of the compound.

[0085] According to one embodiment of the present invention, a culture comprising a strain selected from the group consisting of Streptomyces sp. APA-053 strain (Accession No.: KCTC15538BP) may include a culture medium together with one or more strains selected from the group consisting of Streptomyces sp. APA-053 strain (Accession No.: KCTC15538BP).

[0086] According to one embodiment of the present invention, the culture medium may be any known appropriate medium for culturing the strain, but in order to provide optimal conditions for the production of useful secondary metabolites, the culture medium may be Marine broth (BD), SYP, etc.

[0087] According to one embodiment of the present invention, the medium form of the marine broth may be either a solid medium or a liquid medium, and preferably a liquid medium.

[0088] According to one embodiment of the present invention, the marine broth medium may use soluble starch, yeast extract, peptone, and sea salt.

[0089] According to one embodiment of the present invention, when the medium form of the marine broth is a solid medium, 400 mL may be obtained by adding 4 g of soluble starch, 1.6 g of yeast extract, 0.3 g of peptone, 7.2 g of agar, and 13.9 g of sea salt to 400 mL of distilled water.

[0090] According to one embodiment of the present invention, when the medium form of the marine broth is a liquid medium, 40 g of soluble starch, 16 g of yeast extract, 8 g of peptone, 139 g of sea salt, and 4 L of distilled water may be added based on 4 L.

[0091] According to one embodiment of the present invention, the dried product and / or the concentrated product may be obtained by drying and / or concentrating the culture product by a conventional method.

[0092] One embodiment of the present invention provides a method for producing a compound of formula 1 or a pharmaceutically acceptable salt thereof, comprising the steps of: culturing a Streptomyces genus strain to obtain a culture (S10); extracting the culture with an extraction solvent to obtain an extract (S30); and separating and purifying the extract by chromatography with a developing solvent to obtain a fraction to obtain the compound (S50).

[0093] A method for producing a compound of formula 1 or a pharmaceutically acceptable salt thereof according to one embodiment of the present invention can easily purify a compound having a specific structure, i.e., a compound of formula 1 or a pharmaceutically acceptable salt thereof, and can increase the yield of the compound.

[0094] In this specification, the method for manufacturing the cosmetic composition for skin whitening is omitted to avoid overlapping with that described in the cosmetic composition for skin whitening described above.

[0095] According to one embodiment of the present invention, the method for producing the compound of the above chemical formula 1 or a pharmaceutically acceptable salt thereof includes a step of culturing a strain of the genus Streptomyces to obtain a culture (S10).

[0096] According to one embodiment of the present invention, in the step of culturing the Streptomyces genus strain to obtain a culture, the culture may be obtained by culturing one or more strains selected from the group consisting of Streptomyces sp. APA-053 strain (Accession No.: KCTC15538BP).

[0097] According to one embodiment of the present invention, the culture period for obtaining the culture may be about 4 days or more, preferably 4 to 20 days, and more preferably 6 to 15 days, during which production of effective secondary metabolites is active.

[0098] According to one embodiment of the present invention, a method for producing a compound of formula 1 or a pharmaceutically acceptable salt thereof includes a step (S30) of extracting the culture with an extraction solvent to obtain an extract.

[0099] According to one embodiment of the present invention, a method for extracting 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid represented by the chemical formula 1 from Streptomyces sp. APA-053 strain can use any known microbial extraction method without limitation.

[0100] According to one embodiment of the present invention, in the step of obtaining the extract, that is, the process of extracting 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid represented by the chemical formula 1 from Streptomyces sp. APA-053 strain, the extraction solvent used may be an appropriate solvent known in the art selected according to the characteristics of the microorganism, but may be extracted with an organic solvent, specifically, at least one organic solvent selected from the group consisting of ethyl acetate, methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, chloroform, methylene chloride, hexane, cyclohexane, and dichloromethane, and extraction with an ethyl acetate solvent is preferable. By selecting the solvent used in the step of obtaining the extract from the above, it is possible to exhibit an excellent whitening effect by containing useful secondary metabolites including the compound represented by the chemical formula 1.

[0101] According to one embodiment of the present invention, the extraction may be replaced with a method using an extraction device such as supercritical extraction, high-pressure extraction, or ultrasonic extraction, or a method using celite, polystyrene, or polyamide adsorption resin, but is not limited thereto.

[0102] According to one embodiment of the present invention, the amount of extraction solvent or the extraction temperature during the extraction process may be appropriately changed by a person skilled in the art, but it is preferable to extract by adding the extraction solvent 1 to 3 times the volume of the culture during extraction, and it is preferable to extract at room temperature.

[0103] According to one embodiment of the present invention, the number of extractions in the extraction process is preferably 1 to 3 times, and the vacuum drying is preferably performed using a rotary vacuum evaporator, but is not limited thereto. Furthermore, the temperature during the vacuum drying is preferably 20 to 40°C, and more preferably 30°C, but is not limited thereto.

[0104] According to one embodiment of the present invention, a step (S40) of removing the extraction solvent, i.e., the organic solvent, from the obtained extract may be additionally included between the step (S30) of obtaining the extract and the step (S50) of obtaining the compound. Furthermore, the removal of the organic solvent, i.e., the extraction solvent, may be performed without limitation using any method known in the art.

[0105] According to one embodiment of the present invention, a method for preparing a cosmetic composition for skin whitening includes a step (S50) of obtaining the compound by obtaining fractions by separating and purifying the extract using chromatography as a developing solvent.

[0106] According to one embodiment of the present invention, the chromatography may be any chromatography without limitation based on size, charge, hydrophobicity or affinity, and may preferably be flash chromatography (TLC) or reverse phase liquid chromatography.

[0107] According to one embodiment of the present invention, the developing solvent may be at least one selected from the group consisting of water and alcohol.

[0108] According to one embodiment of the present invention, in the step of obtaining the fraction and obtaining the compound, the fraction may be prepared using a developing solvent (fractionation solvent), and preferably a fraction fractionated with a mixed solvent of water and alcohol (e.g., a lower alcohol having 1 to 3 carbon atoms), more preferably a water / methanol fraction. By selecting the developing solvent (fractionation solvent) from the above, it is possible to contain useful secondary metabolites including the compound represented by Chemical Formula 1 and exhibit an excellent whitening effect.

[0109] According to one embodiment of the present invention, the fraction may be a subfraction obtained by further fractionating the extract into a solution having a concentration of 20 (w / w)%, 40 (w / w)%, 50 (w / w)%, 60 (w / w)%, 70 (w / w)%, 80 (w / w)% or 100 (w / w)% methanol. By using the fraction and / or subfraction as described above, it is possible to exhibit an excellent whitening effect by containing useful secondary metabolites including the compound represented by Chemical Formula 1.

[0110] In addition, the present invention provides a pharmaceutical composition comprising a compound represented by the above chemical formula 1; or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

[0111] In addition, the present invention provides a method for preventing or treating a skin pigmentation disease, comprising administering to a subject in need thereof a compound represented by the above chemical formula 1; or a pharmaceutically acceptable salt thereof.

[0112] In addition, the present invention provides a skin whitening method comprising a step of administering a compound represented by the above chemical formula 1; or a cosmetically acceptable salt thereof, to a subject in need thereof.

[0113] In addition, the present invention provides the use of a compound represented by the above chemical formula 1; or a cosmetically acceptable salt thereof, in the manufacture of a cosmetic for skin whitening.

[0114] In addition, the present invention provides the use of a compound represented by the above chemical formula 1; or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating skin pigmentation diseases.

[0115] In addition, the present invention provides a cosmetic composition comprising a compound represented by the above chemical formula 1 for use in skin whitening; or a cosmetically acceptable salt thereof.

[0116] In addition, the present invention provides a pharmaceutical composition comprising a compound represented by the above chemical formula 1 for use in the prevention or treatment of skin pigmentation diseases; or a pharmaceutically acceptable salt thereof.

[0117] The skin whitening composition according to one embodiment of the present invention is a safe substance because it is derived from a natural product, and can effectively inhibit melanin production, thereby preventing, treating, or improving skin pigmentation, and thus can be usefully used in the manufacture of a composition for developing a whitening pharmaceutical preparation or cosmetic.

[0118] A composition for producing a compound represented by the chemical formula 1 according to one embodiment of the present invention can increase the yield of the effective substance.

[0119] The method for producing a skin whitening composition according to one embodiment of the present invention can easily purify a compound having a specific structure and increase the yield of an effective substance.

[0120] Figure 1 is a photograph showing the morphology of Streptomyces sp. APA-053 strain isolated from moss root marine sediment on a marine solid medium.

[0121] Figures 2a to 2d are graphs measured to identify whether the compound obtained by separation and purification in Example 2 is a compound represented by Chemical Formula 1. Specifically, Figure 2a shows the chemical formula of the compound obtained by separation and purification in Example 2. 1 H NMR results, Figure 2b shows the compound obtained by separation and purification in Example 2. 13 C NMR results, Fig. 2c shows the MS (mass spectrum) of the compound obtained by separation and purification in Example 2, and Fig. 2d shows the UV spectrum results of the compound obtained by separation and purification in Example 2, respectively.

[0122] Figure 3 is a graph showing the results of confirming the concentration-dependent melanin inhibitory effect of the compound represented by the above chemical formula 1 through absorbance measurement.

[0123] Figure 4 is a graph showing the results of confirming intracellular viability according to the addition of the compound represented by the above chemical formula 1.

[0124] Figure 5 is a graph showing the melanin inhibitory effect according to the concentration of 3-indoleacetic acid through absorbance measurement.

[0125] Hereinafter, the present invention will be described in detail using examples. However, the examples according to the present invention may be modified in various ways, and the scope of the present invention is not limited to the examples described below. The examples in this specification are provided to more fully explain the present invention to those of ordinary skill in the art.

[0126] Example 1 (Strain selection and identification)

[0127] Example 1-1: Isolation of strains in sediment

[0128] Sediment samples collected from the moss root marine sediments along the coast of Jukdo Island, Jeju Island were dried in an oven at 65°C for 3 minutes to obtain a dried product. Afterwards, the dried product was suspended in sterilized seawater, diluted to 1 / 2, and inoculated onto ISP4 (Inorganic Salts Starch 4 Agar) solid medium. A single strain produced during cultivation at 27°C was inoculated again onto SYP (Starch Yeast Peptone) SW solid medium to isolate and select a pure strain.

[0129] Figure 1 is a photograph showing the morphology of Streptomyces sp. APA-053 strain isolated from moss root marine sediment on a marine solid medium. The purely isolated and selected APA-053 strain was observed to have the same morphological characteristics as strains belonging to the genus Streptomyces, as shown in Figure 1.

[0130] Example 1-2: Identification of strains

[0131] For species identification of the strain isolated in Example 1-1, 1 ml of the strain cultured in Marine broth liquid medium at 27°C for 4 days was taken, and genomic DNA was extracted using a "Tissue Genomic DNA Isolation kit" (Cosmogenetech co, Ltd., Seoul, South Korea) according to the manufacturer's protocol. For the species analysis, 16S rRNA gene amplification was performed using the 518F and 805R primers.

[0132] The PCR products were purified using a “PCR purification kit” (Cosmogenetech co, Ltd., Seoul, South Korea), and then the base sequence was analyzed using capillary electrophoresis (Applied Biosystems 3730XL).

[0133] The 16S rRNA gene sequence obtained from strain APA-053 was compared with information on previously reported strains using a BLAST search of the GenBank / EMBL / DDBJ database. As a result, the 16S rRNA gene sequence of strain APA-053 showed 99.0% homology with Streptomyces malachitospinus strain NBRC 101004, and the Streptomyces strain APA-053 was determined to be a Streptomyces malachitospinus strain belonging to the genus Streptomyces.

[0134] The 16S rRNA genes base sequence of the above Streptomyces genus APA-053 strain is shown in Table 1 (SEQ ID NO: 1).

[0135]

[0136] Example 2 (Isolation and purification of a compound represented by chemical formula 1)

[0137] Streptomyces sp. APA-053 strain was isolated from a moss root marine sediment sample from the coast of Jukdo, Jeju Island, and a compound represented by the following chemical formula 1 (APA053.243, Marinoprenylindole, 2-(6-(3-methylbut-2-en-1-yl)-1H-indol-3-yl)acetic acid) was isolated and purified.

[0138] [Chemical Formula 1]

[0139]

[0140] Specifically, Streptomyces sp. APA-053 strain was cultured in a 2.5 L culture vessel in 1 L of seawater-containing culture medium (10 g / L starch, 2 g / L yeast extract, 4 g / L peptone) for 7 days at 27°C and 120 rpm (40 L in total). 1 L of ethyl acetate was added to the Streptomyces sp. APA-053 strain culture medium to separate the two layers and obtain an ethyl acetate layer therefrom. Thereafter, the organic solvent was removed from the ethyl acetate layer using a vacuum rotary evaporator, and its extract was obtained.

[0141] The above ethyl acetate extract (743.6 mg) was fractionated with a mixed solvent of water and methanol, and the methanol concentration was fractionated by reverse-phase column chromatography using C-18 silica according to a stepwise gradient from 20 (w / w)% to 100 (w / w)%. The organic solvent was removed from the first fraction fractionated with 80 (w / w)% methanol, and the residue was dissolved in 3 mL of methanol, and high-performance liquid chromatography was performed on the solution to finally obtain a pure compound represented by the above chemical formula 1.

[0142] The conditions of the high-performance liquid chromatography used were as follows.

[0143] Phenomenex Luna C18 (2), 250 mm

[0144] 2.0 mL / min, 5 mm, 100 Å, UV = 254 nm, water (H2O) : acetonitrile (CH3CN) = 52 : 48

[0145] According to the above method, the compound represented by the above chemical formula 1 (3.0 mg) was separated from Streptomyces sp. APA-053 strain at a retention time of 34.4 minutes.

[0146] Example 3 (Identification of the compound represented by Chemical Formula 1)

[0147] In order to confirm whether the compound separated and purified in the above Example 2 is the compound represented by the above Chemical Formula 1, identification was performed using NMR (nuclear magnetic resonance) spectrum, LC-MS (Liquid chromatography - mass spectrometry) spectrum, and ultraviolet spectrum.

[0148] Figures 2a to 2d are graphs measured to identify whether the compound obtained by separation and purification in Example 2 is a compound represented by Chemical Formula 1. Specifically, Figure 2a shows the chemical formula of the compound obtained by separation and purification in Example 2. 1 H NMR results, Figure 2b shows the compound obtained by separation and purification in Example 2. 13 C NMR results, Fig. 2c shows the MS (mass spectrum) of the compound obtained by separation and purification in Example 2, and Fig. 2d shows the UV spectrum results of the compound obtained by separation and purification in Example 2, respectively.

[0149] As a result, as shown in the above Fig. 2a, 1It was confirmed that it contained a benzene ring through the peak between 6.00 ppm and 8.00 ppm in H NMR, and that it contained isoprenylated indole.

[0150] As shown in the above figure 2c, through LC-MS, the mass is [M+H] + It was confirmed that it is 244, and it is a natural product with a molecular weight of 243, and finally, as shown in the above Figures 2b and 2d, 13 Through C NMR and UV spectral analysis, it was confirmed that the compound isolated and purified in Example 2 was the compound represented by the chemical formula 1.

[0151] Example 4 (Measurement of melanin production inhibition ability of a compound represented by chemical formula 1)

[0152] The melanin production inhibitory activity of the compound represented by the above chemical formula 1 was measured using the method of Lee et al. (2013) (J Invest Dermatol. Lee CS, Park M, Han J, Lee JH, Bae IH, Choi H, Son ED, Park YH, Lim KM. 133, 4, 1063-1071).

[0153] B16 melanoma cells (ATCC, USA) were seeded at 1 x 10 4 After dispensing 300 μL of cells / well into a 48-well plate, the cells were cultured overnight in an incubator, and DMEM (Dulbecco's Modified Eagle's Medium) (ATCC, USA) was used as the medium (Con).

[0154] Alpha-MSH (0.1 uM, Sigma-Aldrich, USA), a melanin production stimulant, and its solution in DMSO were injected into the above Con to prepare a negative control (NC). A medium was prepared by simultaneously mixing the compound represented by Chemical Formula 1 at concentrations of 1 ppm (SJ100 1 ppm), 10 ppm (SJ100 10 ppm), 25 ppm (SJ100 25 ppm), 50 ppm (SJ100 50 ppm), and 100 ppm (SJ100 100 ppm) into the negative control group. Arbutin 50 ppm was used as a positive control (PC).

[0155] After removing the medium in the dispensed wells, 450 μL of the prepared medium was added to each well. After 48 hours, the medium was completely removed, 200 μL of 1 N NaOH was added to each well, and the cells were incubated at 60 °C for 1 hour to dissolve melanin from the cells. 100 μL of the dissolved melanin sample was transferred to a 96-well plate, and the absorbance was measured at 405 nm using a microplate reader (Molecular Devices, Sunnyvale, CA, USA).

[0156] From the difference in absorbance, the degree of inhibition of melanin production was calculated as shown in the following mathematical formula 1.

[0157] [Mathematical Formula 1]

[0158] Melanin production inhibition (%) = 1 - (ODS-ODc) / (ODm-ODc) × 100

[0159] (ODS: absorbance of the sample treated with the test substance and alpha-MSH, ODm: absorbance of the sample treated only with alpha-MSH, ODc: absorbance of the negative control group (1N NaOH 100 μL) not treated with the test substance or alpha-MSH)

[0160] Figure 3 is a graph showing the results of confirming the concentration-dependent melanin inhibitory effect of the compound represented by the above chemical formula 1 through absorbance measurement.

[0161] As confirmed in the above Figure 3, the compound represented by the above chemical formula 1 showed a tendency for the melanin production inhibition effect to decrease as the concentration increased to 1 ppm, 10 ppm, 25 ppm, 50 ppm, and 100 ppm.

[0162] According to the above results, it was confirmed that the compound represented by the chemical formula 1 has excellent melanin production inhibition activity at a specific concentration, and can be applied to various diseases and conditions related to skin pigmentation through inhibition of melanin production.

[0163] Example 5 (Confirmation of cytotoxicity of compound represented by chemical formula 1)

[0164] To confirm the cytotoxicity of the compound represented by the above chemical formula 1, cell viability was confirmed. Specifically, B16 cells were cultured at 2x10 4 After suspending the cells / mL, 600 μL was placed in 48 wells and cultured for one day. After treating the material, the cell viability was measured using the MTT assay, which measures mitochondrial activity 48 hours later.

[0165] Figure 4 is a graph showing the results of confirming intracellular viability according to the addition of the compound represented by the above chemical formula 1.

[0166] Referring to the above Figure 4, it was confirmed that addition of the compound represented by the above chemical formula 1 did not significantly affect cell viability and that there was no cytotoxicity.

[0167] Example 6 (Comparison of the melanin production inhibition ability of 3-indoleacetic acid)

[0168] Melanin production inhibition was measured in the same manner as in Example 4, except that 3-indoleacetic acid was used instead of the compound represented by the chemical formula 1 in Example 4.

[0169] Figure 5 is a graph showing the melanin inhibitory effect according to the concentration of 3-indoleacetic acid, a reference example, through absorbance measurement.

[0170] Referring to the above Figure 5, it was confirmed that even when 3-indoleacetic acid was added and its concentration was increased, the melanin concentration remained high, and the melamine production inhibition effect was lower than that of the compound represented by Chemical Formula 1.

[0171] Therefore, the skin whitening composition and the method for preparing the same according to one embodiment of the present invention can inhibit melanin production and improve the skin whitening effect by using a compound having a specific molecular structure derived from a culture of a Streptomyces genus strain as an active ingredient of the skin whitening composition.

[0172] [Explanation of symbols]

[0173] S10: Culture acquisition step

[0174] S30: Extraction step

[0175] S40: Extraction solvent removal step

[0176] S50: Fraction acquisition step

[0177] [Accession number]

[0178] Name of depositor: Korea Research Institute of Bioscience and Biotechnology, Biological Resource Center (KCTC)

[0179] Accession number: KCTC15538BP

[0180] Date of acceptance: 20230804

[0181]

[0182]

Claims

1. A compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof [Chemical Formula 1] .

2. A cosmetic composition for skin whitening comprising a compound represented by the following chemical formula 1; or a cosmetically acceptable salt thereof. [Chemical Formula 1] .

3. In claim 2, A skin whitening cosmetic composition wherein the compound represented by the above chemical formula 1 is derived from Streptomyces sp. APA-053 strain (Accession number: KCTC15538BP).

4. In claim 2, A skin whitening cosmetic composition comprising a compound represented by the above chemical formula 1, which inhibits melanin production in melanoma cells.

5. In claim 2, the skin whitening cosmetic composition has a formulation selected from the group consisting of a solution, an ointment, an external ointment, a cream, an essence, a foam, a toner, a nourishing toner, an emollient, an emollient toner, a pack, a milky lotion, a makeup base, a soap, a cleansing agent, a bath agent, a sunscreen, a sun oil, a suspension, an emulsion, a paste, a gel, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a foundation, a powder foundation, an emulsion foundation, a wax foundation, a patch, and a spray.

6. A pharmaceutical composition for preventing or treating skin pigmentation disease, comprising a compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof. [Chemical Formula 1] .

7. A pharmaceutical composition for preventing or treating a skin pigmentation disease, wherein the skin pigmentation disease in paragraph 6 is any one selected from the group consisting of freckles, liver spots, hyperpigmentation after ultraviolet exposure, hyperpigmentation after inflammation, lentigines, brown spots, and liver spots.

8. A food composition for preventing or improving skin pigmentation disease, comprising a compound represented by the following chemical formula 1; or a food-based acceptable salt thereof. [Chemical Formula 1] .

9. A food composition for preventing or improving skin pigmentation disease, wherein the skin pigmentation disease in paragraph 8 is any one selected from the group consisting of freckles, liver spots, hyperpigmentation after ultraviolet exposure, hyperpigmentation after inflammation, lentigines, brown spots, and liver spots.

10. In a method for producing a compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof, A step of culturing a Streptomyces strain to obtain a culture; A step of extracting the above culture with an extraction solvent to obtain an extract; and A method for producing a compound of formula 1 or a pharmaceutically acceptable salt thereof, comprising the step of obtaining fractions by separating and purifying the extract using a developing solvent through chromatography to obtain the compound: [Chemical Formula 1] .

11. A pharmaceutical composition comprising a compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier. [Chemical Formula 1] .

12. A method for preventing or treating skin pigmentation disease, comprising administering to a subject in need thereof a compound represented by the following chemical formula 1; or a pharmaceutically acceptable salt thereof. [Chemical Formula 1] .

13. Use of a compound represented by the following chemical formula 1 in the manufacture of a drug for treating skin pigmentation diseases; or a pharmaceutically acceptable salt thereof. [Chemical Formula 1] .

14. A pharmaceutical composition comprising a compound represented by the following chemical formula 1 for use in the prevention or treatment of skin pigmentation diseases; or a pharmaceutically acceptable salt thereof. [Chemical Formula 1] .

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