KR102831785B1 - Composition for Promoting Melanin synthesis comprising Sargassum fusiforme extract - Google Patents

Abstract

The present invention relates to a composition for promoting melanin production containing an extract of Sargassum fusiforme . The composition induces blackening of skin and hair by promoting melanin synthesis, and is stable to the skin, so it is very effective as a composition for preventing gray hair and treating vitiligo or as an agent for improving melanin hypopigmentation.

Description

Composition for promoting melanin synthesis comprising Sargassum fusiforme extract

The present invention relates to a composition containing a Sargassum fusiforme extract for promoting melanin production. More specifically, the present invention relates to a composition containing a Sargassum fusiforme extract as an effective ingredient for preventing and improving gray hair, vitiligo, and hypopigmentation by promoting melanin synthesis and inducing darkening while being safe for the skin.

Melanin, a pigment produced by melanocytes in the basal layer of the epidermis of the human skin, is a phenolic polymer that is a complex of black pigment and protein. It determines human skin color depending on its concentration and distribution, and also plays a role in protecting our skin from ultraviolet rays. If melanin is excessively produced due to causes such as ultraviolet rays and hormones, it causes pigmentation such as blemishes and freckles, but conversely, if melanin production is suppressed, it can become a risk factor for skin cancer due to hypopigmentation and skin lesions such as vitiligo can appear. Therefore, research on inducing darkening by promoting melanin production is just as important as research on whitening that suppresses melanin production.

In hair, gray hair occurs naturally as aging progresses, but it is not only unsightly, but also causes difficulties in social activities, so many people try to hide it by temporarily dyeing their gray hair black, and so much research is being done on hair dyes. However, hair dyes are temporary methods, so they require periodic use because gray hair grows back, and they can damage the skin because they contain oxidizing agents. Therefore, research on melanin production accelerators is also actively being conducted to fundamentally prevent or improve gray hair.

Gray hair can also be caused by external stress. The scalp, like the skin, protects the body from the external environment and excretes harmful substances and waste products in the body. If oxidative stress accumulates in the hair follicles due to external substances, not only will the hair become thinner, but it can also affect melanin formation, causing gray hair.

Taking this into account, the inventors of the present invention sought to find a substance that could alleviate the problems of hypopigmentation and gray hair, and selected tangle as such a substance.

Sargassum fusiforme is an edible seaweed belonging to the family Sargassum, a perennial seaweed that grows in the lower intertidal zones of Korea, Japan, and China. It grows in colonies on gently sloping reefs or in areas with gentle waves, has fibrous roots, and has a conical stem structure. Sargassum is a seaweed that starts growing in winter and is in season from February to July. In Korea, it is mainly cultivated in the southern coast and Jeju Island. Sargassum is a seaweed that is rich in nutrients and contains a large amount of various physiologically active substances, so it is called the elixir of the sea, and has been eaten a lot since ancient times. In Jeju, it was also eaten a lot as a famine relief food during famine. It is rich in dietary fiber, vitamins, and fucoidan, and is popular as an anticancer food because it has excellent effects of excreting heavy metals from the body through alginic acid.

Meanwhile, Korean Patent Publication No. 10-2021-0106661 and Japanese Patent Publication No. 2011-79768 disclose that hot water extract of thuja inhibits melanin production.

In contrast, the inventors of the present invention have discovered that when the extraction method of the extract of the sea tangle is changed, unlike the above patent documents, the sea tangle is safe for the skin and promotes melanin synthesis to induce blackening, thereby being very effective in preventing and improving gray hair, vitiligo, and hypopigmentation, and thus completed the present invention.

One object of the present invention is to provide a composition for promoting melanin production containing a turmeric extract.

Another object of the present invention is to provide a method for producing a tangle extract having the effect of promoting melanin production.

Another object of the present invention is to provide a cosmetic composition for preventing or improving gray hair, vitiligo, or hypopigmentation, comprising the composition for promoting melanin production.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating gray hair, vitiligo, or hypopigmentation, comprising the composition for promoting melanin production.

Another object of the present invention is to provide a composition for external application to the skin for preventing, improving or treating gray hair, vitiligo or hypopigmentation, comprising the composition for promoting melanin production.

Another object of the present invention is to provide a food composition for preventing or improving gray hair, vitiligo, or hypopigmentation, comprising the composition for promoting melanin production.

Another object of the present invention is to provide a method for preventing, improving, or treating gray hair, vitiligo, or hypopigmentation in a subject by administering to the subject a composition for promoting melanin production.

In one embodiment, the present invention provides a composition for promoting melanin production comprising a sesame extract.

In the present invention, " Sargassum fusiforme " is an edible seaweed belonging to the family Sargassum, a brown algae, and is a perennial seaweed that lives in the lower intertidal zone of Korea, Japan, and China.

The present invention relates to a method for producing a tangle extract, comprising: (S10) a step of first extracting tangle with water, an organic solvent, or a mixed solvent thereof; (S20) a step of removing alginic acid from the tangle extract and drying it; (S30) a step of second extracting the tangle extract from which the alginic acid has been removed and dried by precipitating the tangle extract in an organic solvent; and (S40) a step of obtaining a supernatant of the tangle extract.

Hereinafter, the method for producing the thuja extract of the present invention will be specifically described with reference to FIG. 1.

(S10) Step is the step of first extracting the husk with water, an organic solvent, or a mixed solvent thereof.

The above organic solvent may be a polar or nonpolar solvent, preferably a lower alcohol having 1 to 4 carbon atoms (such as ethanol, methanol, propanol or butanol), and the first extraction may be performed using various extraction methods, such as hot water extraction, cold extraction, ultrasonic extraction, or reflux cooling extraction, with these solvents, preferably a hot water extraction method.

As a specific example, the first extraction may use a heat extraction method in which dried tangle powder is immersed in water and heated to 90 to 120°C, preferably 100°C, to perform extraction. Meanwhile, the extraction time is preferably 10 minutes to 4 hours, more preferably 60 minutes to 3 hours, and most preferably 2 hours, but is not limited thereto. In addition, the number of extractions is preferably 1 to 5 times, more preferably 1 to 3 times of repeated extraction, and most preferably 2 times, but is not limited thereto.

Meanwhile, a step of drying the seaweed may be included before the step (S10), and the drying is preferably done in the shade.

Step (S20) is a step of removing alginic acid from the primary extract of the above-mentioned tangle and drying it.

The removal of the above alginic acid is not necessarily limited to this, but may be accomplished by using a method such as coagulation and removal through a cross-linking reaction with a calcium salt.

The above drying can be accomplished by a conventional drying method, such as freeze drying, reduced pressure drying, vacuum drying, boiling drying, or spray drying.

(S30) Step is a step of secondary extraction by precipitating the first extract of the dried seaweed from which the alginate has been removed in an organic solvent.

The above organic solvent is a lower alcohol having 1 to 4 carbon atoms (e.g. ethanol, methanol, propanol or butanol), preferably ethanol or methanol, and most preferably ethanol.

The above precipitation can be carried out at room temperature for 60 minutes to 4 hours, preferably 60 minutes to 3 hours, more preferably 90 minutes to 150 minutes, and most preferably 120 minutes.

As a specific example, the first extract of the above-mentioned tangle is dried after removing alginic acid, and then the first extract is immersed in a 60 to 80% ethanol solution, preferably a 65 to 75% ethanol solution, more preferably a 70% ethanol solution, and the process can be performed at room temperature for 60 to 240 minutes, preferably 60 minutes to 3 hours, more preferably 90 to 150 minutes, and most preferably 120 minutes.

(S40) Step is a step of obtaining a supernatant of the secondary extract of the above-mentioned thuja.

The above supernatant is a supernatant generated by the precipitation of the above (S30) step, and will be understood to include all of the solids of the supernatant, dilutions thereof, and concentrates thereof.

The extract of the present invention manufactured by the above method has the effect of promoting the production and blackening of melanin in hair and skin with reduced melanin pigment.

Accordingly, the present invention provides, as another aspect, a method for producing a thuja extract that promotes the production and blackening of melanin in hair and skin.

In one specific embodiment, it was confirmed that the thuja extract according to the present invention has the effect of significantly promoting melanin production in human melanocytes and B16 melanoma cells.

In another specific embodiment, it was confirmed that the extract of the present invention has the effect of promoting melanin production in experiments using 3D skin and human hair follicle culture.

Therefore, the extract of the present invention promotes melanin synthesis and has high stability on the skin, so it can be usefully used for skin darkening, prevention of gray hair, treatment and prevention of vitiligo, and can be usefully used in cosmetics, medicines and foods for inducing darkening.

In the composition of the present invention, the effective ingredient of the tilia extract may be included in an amount of 0.0001 to 10.0 wt%, preferably 0.001 to 10 wt%, based on the total weight of the composition. If the content of the tilia extract is less than 0.0001 wt%, the effect of preventing, improving, or treating the darkening effect of skin and hair and vitiligo or hypopigmentation of the skin is too weak, and if it exceeds 10.0 wt%, there is a problem that the increase in effect according to the increase in the content is insignificant and the stability of the formulation is not secured.

In another aspect, the present invention provides a cosmetic composition for improving or preventing skin darkening effects, such as white hair, vitiligo or hypopigmentation, comprising a composition containing the extract of the above-mentioned thuja as an effective ingredient.

The prevention or improvement effects of gray hair, vitiligo and hypopigmentation due to the melanin production-promoting effect of the above-mentioned extract of the plant are as described above.

In the present invention, the term "prevention" or "suppression" means reducing the occurrence of a disease or condition in a subject who has not been diagnosed as having the disease or condition, but is prone to developing such disease or condition.

As used herein, the term “treatment” or “improvement” means (a) inhibition of the development of a disease or condition, (b) alleviation of the disease or condition, and (c) elimination of the disease or condition in a subject.

As used herein, the term “subject” refers to a mammal, including a human, monkey, cow, horse, pig, sheep, dog, cat, rat, mouse, chimpanzee, etc., having a disease whose symptoms can be improved by administering the composition of the present invention.

The above composition contains, in addition to the extract of thuja as an effective ingredient, ingredients commonly used in cosmetic compositions, and includes conventional auxiliary agents and carriers such as antioxidants, solubilizers, stabilizers, vitamins, pigments, and fragrances.

The cosmetic composition of the present invention can be manufactured into any formulation generally manufactured in the art, and for example, can be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray, but is not limited thereto. In addition, it can be manufactured into the formulation of a nourishing cream, astringent toner, flexible toner, lotion, essence, shampoo, nourishing gel, or massage cream.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane/butane or dimethyl ether may be additionally included.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizer or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, etc. can be used as carrier components.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl amidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester may be used as a carrier component.

In particular, the cosmetic composition for improving or preventing gray hair, vitiligo or hypopigmentation of the present invention provides a hair care cosmetic composition that promotes melanin production in the hair and suppresses gray hair when applied to the hair.

The above-mentioned cosmetic compositions for hair care include, for example, hair shampoo, hair rinse, hair conditioner, hair treatment, hair tonic, scalp treatment, hair lotion, hair cream, hair mist, hair ampoule, hair pack, hair essence, hair oil, scalp scaler, hair nourishing toner, and hair dye.

The cosmetic composition of the present invention may contain a tilia extract as an active ingredient in an amount of 0.0001 to 10.0 wt%, preferably 0.001 to 10 wt%, based on the weight of the total composition. If the content of the tilia extract is less than 0.0001 wt%, the effect of preventing, improving, or treating skin and hair blackening and vitiligo or hypopigmentation of the skin is too weak, and if it exceeds 10.0 wt%, there is a problem that the increase in effect due to the increase in the content is insignificant and the stability of the formulation is not secured.

In another aspect, the present invention provides a pharmaceutical composition comprising the composition for preventing or treating skin darkening, i.e., white hair, vitiligo or hypopigmentation.

The preventive or therapeutic effects of the above-mentioned extract of the plant on white hair, vitiligo, and hypopigmentation due to the melanin production-promoting effect are as described above.

The term "pharmaceutical composition" of the present invention means a composition manufactured for the purpose of preventing or treating a disease, and each may be formulated and used in various forms according to conventional methods. For example, it may be formulated as an oral formulation such as a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, etc., and may be formulated and used as a parenteral formulation using a diluent or excipient such as a lubricant, a wetting agent, a flavoring agent, an emulsifier, a suspending agent, a preservative, a surfactant, etc. In addition, it may be formulated and used in the form of an external preparation, a suppository, an external skin preparation, and a sterile injection solution. As a specific example, the composition of the present invention may be used in an oral administration manner as an oral formulation. As another specific example, the composition of the present invention may be used by a method such as directly applying or spraying it on hair and skin with reduced melanin pigment.

In addition, depending on each formulation, it can be manufactured by additionally including a pharmaceutically acceptable carrier, such as a buffer, an analgesic, a solubilizer, an isotonic agent, a stabilizer, a base, etc. known in the art.

The term "pharmaceutically acceptable carrier" used in the present invention may mean a carrier or diluent that does not stimulate a living organism and does not inhibit the biological activity and properties of the active ingredient to be injected. The type of the carrier usable in the present invention is not particularly limited, and any carrier that is commonly used in the relevant technical field and is pharmaceutically acceptable may be used. Non-limiting examples of the carrier include saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and the like. These may be used alone or in combination of two or more. The carrier may be a non-naturally occurring carrier.

The composition of the present invention can be administered in a pharmaceutically effective amount. The pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and not causing side effects, and the effective dosage level can be determined according to factors including the patient's health condition, the type and severity of the disease, the activity of the drug, the sensitivity to the drug, the administration method, the administration time, the administration route and the excretion rate, the treatment period, the drugs used in combination or simultaneously, and other factors well known in the medical field. Specifically, the dosage is generally 0.01 mg to 5000 mg per 1 kg of the body weight of the administered subject per day, and may be administered once or several times a day at regular intervals depending on the judgment of a doctor or pharmacist, but is not limited thereto.

The pharmaceutical composition of the present invention may contain a tilia extract as an active ingredient in an amount of 0.0001 to 10.0 wt%, preferably 0.001 to 10 wt%, based on the weight of the total composition. If the content of the tilia extract is less than 0.0001 wt%, the effect of preventing, improving, or treating darkening of the skin and hair and vitiligo or hypopigmentation of the skin is too weak, and if it exceeds 10.0 wt%, there is a problem that the increase in effect according to the increase in the content is insignificant and the stability of the formulation is not secured.

In addition, the pharmaceutical composition of the present invention can be used alone or in combination with or as an appropriate set of other pharmaceutically active compounds that exhibit a preventive or therapeutic effect on gray hair, vitiligo, or hypopigmentation.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And may be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without causing side effects by considering all of the above factors, and this can be easily determined by those skilled in the art.

In the present invention, the term "administration" means introducing the pharmaceutical composition of the present invention into a subject by any appropriate method, and the route of administration of the composition of the present invention may be through various routes, such as oral or parenteral, as long as it can reach the target tissue.

The method of administration of the pharmaceutical composition according to the present invention is not particularly limited, and may follow a method commonly used in the relevant technical field. As a non-limiting example of the above-mentioned method of administration, the composition may be administered orally or parenterally. Specifically, the composition of the present invention may be used by a method such as directly applying or spreading it on a part of the hair or skin where melanin pigment is reduced.

The pharmaceutical composition according to the present invention can be prepared in various dosage forms depending on the intended administration method.

The frequency of administration of the composition of the present invention is not particularly limited, but may be administered once a day or administered several times in divided doses.

In another aspect, the present invention provides a composition for external application to the skin for preventing, improving or treating gray hair, vitiligo or hypopigmentation, comprising a composition for promoting melanin production containing the extract of the above-mentioned thuja.

The prevention or improvement effects of gray hair, vitiligo and hypopigmentation due to the melanin production-promoting effect of the above-mentioned extract of the plant are as described above.

The external skin composition according to the present invention can be formulated containing a cosmetically or dermatologically acceptable medium or base. It can be provided in any formulation suitable for topical application, for example, in the form of a solution, a gel, a solid, a pasty anhydrous product, an emulsion obtained by dispersing an oil phase in an aqueous phase, a suspension, a microemulsion, a microcapsule, a microgranule, or a vesicular dispersion of ionic (liposomes) and nonionic types, or in the form of a cream, a skin, a lotion, a powder, an ointment, a spray, a pack or a concealer stick. It can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions can be prepared according to conventional methods in the art.

In addition, the skin external composition according to the present invention may contain adjuvants commonly used in the fields of cosmetology or dermatology, such as fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestering agents, chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetics. The adjuvants are introduced in an amount commonly used in the fields of cosmetology or dermatology.

The thuja extract included in the above composition is not particularly limited thereto, but may be contained in an amount of 0.0001 to 10.0 wt%, more preferably 0.001 to 10 wt%, based on the total weight of the composition. If the content of the thuja extract is less than 0.0001 wt%, the effect of preventing, improving, or treating the darkening effect of skin and hair and vitiligo or hypopigmentation of the skin is too weak, and if it exceeds 10.0 wt%, there is a problem that the increase in effect according to the increase in the content is insignificant and the stability of the formulation is not secured.

In another aspect, the present invention provides a food composition for preventing or improving diseases related to white hair, vitiligo, and hypopigmentation, comprising a composition for promoting melanin production containing the extract of the above-mentioned thuja.

The prevention or improvement effects of gray hair, vitiligo and hypopigmentation due to the melanin production-promoting effect of the above-mentioned extract of the plant are as described above.

When the composition of the present invention is used as a food additive, the thuja extract can be added as it is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The mixing amount of the effective ingredient can be appropriately determined according to the purpose of use (prevention, health, or therapeutic treatment), and a food-related acceptable food auxiliary additive can be additionally included. The thuja extract of the present invention has been used for food, and its safety has been confirmed through human safety tests, so there is no significant limitation on the mixing amount.

The food composition of the present invention can include all foods in the conventional sense, and can be used interchangeably with terms known in the art, such as functional food and health functional food.

There is no limitation on the type of food in which the composition of the present invention can be used for the prevention or improvement of diseases related to the prevention or improvement of white hair, vitiligo, and hypopigmentation through the promotion of melanin production. In addition, a composition comprising the extract of the present invention as an active ingredient can be prepared by mixing other appropriate auxiliary ingredients that can be contained in foods and known additives according to the selection of a person skilled in the art. Examples of foods to which the composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and the like can be prepared by adding the extract according to the present invention to juice, tea, jelly, and juice made using the extract as a main ingredient.

In addition, foods applicable to the present invention may include all foods, such as special nutritional foods (e.g., formulated milk, infant/toddler food, etc.), processed meat products, fish products, tofu, starch jelly, noodles (e.g., ramen, noodles, etc.), health supplements, seasoned foods (e.g., soy sauce, soybean paste, red pepper paste, mixed sauce, etc.), sauces, confectionery (e.g., snacks), processed dairy products (e.g., fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchi types, pickled vegetables, etc.), beverages (e.g., fruit and vegetable beverages, soy milk, fermented beverages, etc.), natural seasonings (e.g., ramen soup, etc.).

The thuja extract included in the above composition is not particularly limited thereto, but is contained in an amount of 0.0001 to 10 wt%, more preferably 0.001 to 10 wt%, based on the total weight of the composition. If the content of the thuja extract is less than 0.0001 wt%, the effect of preventing, improving, or treating the darkening effect of skin and hair and vitiligo or hypopigmentation of the skin is too weak, and if it exceeds 10.0 wt%, there is a problem that the increase in effect according to the increase in the content is insignificant and the stability of the formulation is not secured.

When the health functional food composition of the present invention is used in the form of a beverage, it may contain various sweeteners, flavoring agents, or natural carbohydrates as additional ingredients, like conventional beverages. In addition to the above, the health functional 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 regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, and the like. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks.

In another aspect, the present invention provides a method for preventing, improving or treating diseases related to white hair, vitiligo or hypopigmentation, comprising a composition comprising the extract of the above-mentioned thuja.

The prevention or treatment effects of gray hair, vitiligo and hypopigmentation due to the melanin production-promoting effect of the above-mentioned extract of the plant are as described above.

The term "subject" used in the present invention may mean any animal, including a human, whose hair or skin color has become white or may become white in whole or in part due to a decrease in melanin pigment. The animal may be, but is not limited to, a mammal, such as a cow, horse, sheep, pig, goat, camel, antelope, dog, or cat, which requires treatment for symptoms similar thereto, as well as a human.

The above preventive or therapeutic method of the present invention may specifically include a step of administering the composition in a pharmaceutically effective amount to an individual whose hair or skin in whole or in part has become white or is likely to become white due to a decrease in melanin pigment. The method of administration is as described above.

The above prevention may mean any act of inhibiting or delaying the onset of symptoms and diseases resulting from a decrease in melanin pigment, such as gray hair, vitiligo, and hypopigmentation, by administering to a subject a composition for promoting melanin production according to the present invention.

The above treatment may mean any act of administering the composition of the present invention to a subject suspected of having symptoms or diseases due to a decrease in melanin pigment, thereby improving or beneficially aggravating the symptoms or symptoms of the disease due to a decrease in melanin pigment.

The pharmaceutical composition containing the above-mentioned extract of the plant is as described above.

The composition of the present invention may contain, for administration, a pharmaceutically acceptable carrier, excipient or diluent in addition to the effective ingredients described above. The carrier, excipient and diluent may include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The appropriate dosage of the composition of the present invention varies depending on the patient's condition and weight, the degree of the disease, the drug form, and the time, but can be appropriately selected by a person skilled in the art. Specifically, the dosage of the above-mentioned thuja extract can be 1 to 500 mg/kg.

Meanwhile, the effective ingredient of the present invention, the thuja extract, has not only been used for food, but has also been found to be harmless to the human body as a result of a skin cumulative irritation test. Therefore, the thuja extract of the present invention has almost no toxicity or side effects, so it can be used safely even for long-term use, and can be safely used in cosmetics, medicines, foods, and external skin preparations.

The composition for promoting melanin production containing the extract of the present invention as an active ingredient can be effectively used as an active ingredient of cosmetics, medicines, foods, and external skin agents for preventing, improving, or treating gray hair, gray hair and hypopigmentation of the skin, etc. by inducing blackening of hair and skin due to a decrease in melanin pigment.

Figure 1 is a diagram schematically illustrating a method for producing a thuja extract according to one embodiment of the present invention.
Figure 2 is a drawing showing the effect of a seaweed extract on promoting melanin synthesis in human melanocytes.
Figure 3 is a drawing showing the effect of the extract of Houttuynia cordata on promoting melanin synthesis in B16 melanoma cells.

Hereinafter, in order to help understand the present invention, examples and the like will be described in detail. However, the examples according to the present invention may be modified in various different forms, and the scope of the present invention should not be construed as being limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to a person having average knowledge in the art.

Example 1: Preparation of Tooth Extract

Jeju seaweed was collected, washed thoroughly to completely remove foreign substances and impurities, shade-dried at 20 to 35°C, and extracted with 100°C water for 2 hours. Calcium chloride was added to the hot water extract of seaweed to a final concentration of 2%, and alginic acid was gelled through a cross-linking reaction and removed by filtration. The seaweed extract from which alginic acid had been removed was freeze-dried and then precipitated in a 70% ethanol solution for 2 hours. After precipitation, only the supernatant was recovered, filtered through a 5 μm filter, concentrated in a reduced pressure concentrator, and then freeze-dried to prepare a seaweed extract powder.

Example 2: Cytotoxicity test of the extract of the plant

In order to confirm the cytotoxicity of the extract of the present invention, the following experiment was performed using human melanocytes and human keratinocytes.

Human melanocytes were seeded at 5 x 10 ³ per well in a 96-well plate with M254 medium containing Human Melanocytes Growth Supplement (HMGS), and human keratinocytes were seeded at 5 x 10 3 per well in a 96-well plate with EpiLife™ medium containing Human Keratinocytes Growth Supplement (HKGS). The cells were then cultured at 5% CO ₂ and ³⁷ °C until approximately 80% or more of the cells were attached to the bottom of the well. The medium was then removed, replaced with each medium in which the samples were diluted at various concentrations ranging from 10 ppm to 150 ppm, and then cultured at 5% CO ₂ and 37°C for 3 days. After 3 days, the culture medium was removed and the MTT reagent dissolved at 1 mg/㎖ was diluted 10-fold with each medium and treated, and then cultured for 4 hours. After 4 hours, the culture medium containing the MTT reagent was removed, 100 ㎕ of DMSO was added per well, and the formazan produced in the cells was dissolved and the absorbance was measured at 540 nm. The cell viability was calculated by comparing it with the control value that was not treated with the extract of Houttuynia cordata.

As a result, as shown in Table 1, it was confirmed that there was no cytotoxicity in all treatment groups of the thuja extract.

Sample Treatment concentration (ppm) human melanocytes human keratinocytes Cell viability (%) control - 100 100 Tofu extract 10 102.3 101.2 50 102.7 99.8 100 100.3 101.1 150 100.4 104.2

Example 3: Effect of stimulating melanin production by extracts of tangle in human melanocytes

In order to determine the effect of promoting melanin production by the extract of the present invention, the following experiment was performed using human melanocytes.

Human melanocytes were seeded at 1.5 x 10 ⁵ per well in a 6-well plate with M254 medium containing HMGS (Human Melanocytes Growth Supplement), and cultured under 5% CO ₂ and 37°C until the cells were attached to the bottom of the well by about 80%. After culture, the medium was removed and replaced with a medium in which the sample was diluted to an appropriate concentration, and cultured under 5% CO ₂ and 37°C for 5 days. The concentration range of the Hizikia extract was determined to be 50 ppm, 100 ppm, and 150 ppm without cytotoxicity according to the above Example 2. The cells from which the medium was removed were washed with PBS (phosphated buffer saline), and the cells were harvested. The harvested cells were counted for the cell number using a hemocytometer, and then centrifuged at 10,000 to 13,000 rpm for 10 minutes to remove the supernatant, and a pellet was obtained. After drying the obtained cell pellet at 60℃, 100 ㎕ of 1 M sodium hydroxide solution containing 10% DMSO was added to obtain an intracellular melanin solution in a 60℃ constant temperature bath. The absorbance of this solution at 450 nm was measured using a microplate reader to obtain the amount of melanin per cell number. The results are shown in Table 2 and Fig. 2 below.

Sample Treatment concentration (ppm) Melanin production promotion rate (%) control - 100 Tofu extract 50 138 100 148.9 150 154.5

As described in Table 2 and Figure 2 above, the extract of the present invention was observed to have a higher melanin production promotion rate than the control group in a concentration-dependent manner.

Example 4: Effect of stimulating melanin production by safflower extract in B16 melanoma cells

In order to determine the melanin production promoting effect of the extract of the present invention, the following experiment was performed using B16 melanoma cells.

B16 melanoma cells were seeded at 1x105 per well in a 6-well plate with DMEM medium containing 10% FBS (fetal bovine serum) and cultured under ⁵ % _CO2 and 37℃ until the cells were attached to the bottom of the well by about 80%. After culture, the medium was removed, the sample was replaced with a medium diluted to an appropriate concentration, and cultured under 5% _CO2 and 37℃ for 3 days. The concentration range of the Hizikia extract was determined to be 10 ppm, 50 ppm, and 100 ppm without cytotoxicity. The cells from which the medium was removed were washed with PBS (phosphate buffer saline), and the cells were harvested. The harvested cells were counted using a hemocytometer, and then centrifuged at 10,000 to 13,000 rpm for 10 minutes to remove the supernatant and obtain a pellet. After drying the obtained cell pellet at 60℃, 100 ㎕ of 1 M sodium hydroxide solution containing 10% DMSO was added to obtain intracellular melanin in a 60℃ constant temperature bath. The absorbance of this solution at 450 nm was measured using a microplate reader to obtain the amount of melanin per cell number. The results are shown in Table 3 and Fig. 3 below.

Sample Treatment concentration (ppm) Melanin production promotion rate (%) control - 100 Tofu extract 10 142.3 50 173 100 198.5

As described in Table 3 and FIG. 3 above, the thuja extract according to the present invention showed a higher rate of promotion of melanin production in a concentration-dependent manner than the control group, and it was observed that melanin production was promoted approximately twice when 100 ppm of the thuja extract was treated.

Example 5: Effect of stimulating melanin production by extracts of turmeric in 3D skin

In order to investigate the effect of promoting melanin production by the extract of the present invention, the following experiment was performed using 3D skin. The 3D skin was cultured in a Maintenance medium at 37°C, 5% _CO2 in a 12-well plate. After replacing the medium, the extract of the thuja prepared in Example 1 was treated at non-cytotoxic concentrations of 50 ppm and 100 ppm. After replacing the medium three times with the medium treated with the extract of the thuja at the appropriate concentration for 10 days, the medium was cultured under 5% _CO2 and 37°C. The cells from which the medium was removed were washed with PBS (phosphate buffer saline), and the cells were harvested. The supernatant was removed by centrifugation at 10,000 to 13,000 rpm for 10 minutes, and a pellet was obtained. After drying the obtained cell pellet at 60℃, 100 ㎕ of 1 M sodium hydroxide solution containing 10% DMSO was added to obtain intracellular melanin in a 60℃ thermostatic bath. The absorbance of this solution at 450 nm was measured using a microplate reader to obtain the amount of melanin per cell number. The results are shown in Table 4 below.

Sample Treatment concentration (ppm) Melanin production promotion rate (%) control - 100 Tofu extract 50 134 100 141

As described in Table 4 above, the extract of the thuja according to the present invention was observed to have a higher melanin production promotion rate in a concentration-dependent manner than the control group.

Example 6: Effect of the extract of the plant on melanin production through human hair follicle culture

The melanin production-promoting effect of the H. chinensis extract prepared in Example 1 was measured through human hair follicle culture. Specifically, hair follicles obtained from female donors were cultured at 37°C for 7 days in E. William's medium containing 50 U/mL penicillin, 50 μg/mL streptomycin, 2 mM L-glutamine, 10 μg/mL insulin, and 0.004 μg/mL hydrocortisone, and treated with the H. chinensis extract. After fixing with formaldehyde, the cells were dehydrated, fixed in paraffin, stained with Fontana-Masson staining solution, and cross-sections were cut to a thickness of 5 μm and observed under a Nikon E400 microscope. The melanin content of the hair follicles was quantitatively measured using Image J software for melanin in the hair follicle area, and shown in Table 5. As a result, compared to the untreated negative control group, a 39% increase in melanin content was observed in the group treated with 100 ppm of the tofu extract.

Sample Treatment concentration (ppm) Melanin production promotion rate (%) control - 100 Tofu extract 50 122 100 139

Example 7: Safety test of the extract of turmeric on human skin

In order to verify whether the extract of the present invention is safe for human skin, a skin safety verification experiment was performed. For this purpose, a skin cumulative irritation test was performed. Formulations containing 0.1%, 0.5%, and 1% of the extract of the present invention were manufactured, and these were used to apply patches to the upper arm area of 30 healthy adults every other day for a total of 9 24-hour cumulative patches to measure whether the extract of the present invention irritates the skin. A Finn chamber (Finn chamber, Epitest Ltd, Finland) was used for the patch application. 15 ㎕ of each of the above external skin agents was dropped into the chamber, and then the patch application was performed. The degree of the reaction shown on the skin each time was scored using the following mathematical formula 1, and the results are shown in Table 6 below.

[Mathematical formula 1]

Average Response = [[{(Response Index × Response) / (Total Number of Examiners × Highest Score (4 points))}] × 100] ÷ Number of Examinations (9 times)

In the reactivity scale, ± is given 1 point, + is given 2 points, and ++ is given 4 points, and a composition is judged to be safe when the average reactivity scale is less than 3.

Test substance Number of subjects who responded Average Response 1 week 2 weeks 3 weeks 1st 2nd 3rd 4th 5th 6th 7th 8th 9th ± ＋ ＋＋ ± ＋ ＋＋ ± ＋ ＋＋ ± ＋ ＋＋ ± ＋ ＋＋ ± ＋ ＋＋ ± ＋ ＋＋ ± ＋ ＋＋ ± ＋ ＋＋ Control group
(Squalene) 0 - - 0 - - - - - - - - - - - - - - - - - - - - - - - 0.00 Hot
extract
(0.1%)
[Test group 1] 0 - - 0 - - - - - - - - - - - - - - - - - - - - - - - 0.00 Hot
extract
(0.5%) [Test group 2] 0 - - 0 - - - - - - - - - - - - - - - - - - - - - - - 0.00 Tofu extract
(1%)
[Test group 3] 0 - - 0 - - - - - - - - - - - - - - - - - - - - - - - 0.00

In Table 5 above, the number of people corresponding to ±, +, and ++ in test groups 1, 2, and 3 was all 0, and the average reaction rate was also 0.00. Therefore, the average reaction rate was 3 or less, and the extract of the thuja was determined to be a safe substance for human skin that does not show a distinct cumulative irritation pattern.

The composition of the present invention can be prepared in the following formulations, but is not limited thereto.

Formulation Example 1: Cosmetic Formulation

1-1. Flexible toner

As shown in Table 7 below, a flexible toner containing a turmeric extract as an effective ingredient was manufactured according to a conventional method.

ingredient weight% Tofu extract 0.01 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 Ethanol 10.0 Triethanolamine 0.1 Preservatives, trace colors, trace flavors and trace purified water 82.79 aggregate 100.0

1-2. Nourishing Toner

As shown in Table 8 below, a nutritional toner containing a turmeric extract as an effective ingredient was manufactured according to a conventional method.

ingredient weight% Tofu extract 0.01 beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 5.0 Squalane 5.0 Caprylic/Capric Triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservatives, trace colors, trace flavors and trace purified water 69.69 aggregate 100.0

1-3. Nourishing cream

A nutritional cream containing a turmeric extract as an active ingredient was manufactured according to a conventional method, as shown in Table 9 below.

ingredient weight% Tofu extract 0.01 beeswax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 10.0 Squalane 5.0 Caprylic/Capric Triglyceride 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, trace colors, trace flavors and trace purified water 56.79 aggregate 100.0

1-4. Massage Cream

A massage cream containing a perilla extract as an active ingredient was manufactured according to a conventional method, as shown in Table 10 below.

ingredient weight% Tofu extract 0.01 beeswax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Caprylic/Capric Triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, trace colors, trace flavors and trace purified water 27.49 aggregate 100.0

1-5. Pack

A pack containing a turmeric extract as an effective ingredient, as shown in Table 11 below, was manufactured according to a conventional method.

ingredient weight% Tofu extract 0.01 polyvinyl alcohol 13.0 Sodium carboxymethyl cellulose 0.2 Allantoin 0.1 Ethanol 5.0 Nonylphenyl ether 0.3 Preservatives, trace colors, trace flavors and trace purified water 81.39 aggregate 100.0

1-6. Shampoo

A shampoo containing a turmeric extract as an active ingredient was manufactured according to a conventional method, as shown in Table 12 below.

ingredient weight% Tofu extract 0.01 TEA-Cocoyl Glutamate 10.0 Cocamidopropyl Betaine 20.0 glycerin 5.0 Panthenol 0.05 Niacinamide 0.2 Xanthan gum 1.0 Preservatives, trace colors, trace flavors and trace purified water 63.74 aggregate 100.0

1-7. Hair Serum

A hair serum containing a turmeric extract as an active ingredient was manufactured according to a conventional method, as shown in Table 13 below.

ingredient weight% Tofu extract 0.01 menthol 0.1 Polysorbate 20 0.5 Ethanol 5.0 Panthenol 0.1 Niacinamide 0.3 Pyridoxine HCL 0.1 Dipropylene glycol 2.0 Preservatives, trace colors, trace flavors and trace purified water 91.89 aggregate 100.0

Preparation Example 2: Pharmaceutical preparation

2-1. Manufacturing of Sanje

ingredient Content (g) Tofu extract 2 Lactose 1

The above ingredients were mixed and filled into a sealed bag to prepare a powdered preparation.

2-2. Manufacturing of tablets

ingredient Content (mg) Tofu extract 100 Corn starch 100 Lactose 100 Magnesium stearate 2

After mixing the above ingredients, tablets were manufactured by pressing them according to the usual method for manufacturing tablets.

2-3. Manufacturing of capsules

ingredient Content (mg) Tofu extract 100 Corn starch 100 Lactose 100 Magnesium stearate 2

After mixing the above ingredients, the mixture was filled into a gelatin capsule according to a conventional capsule manufacturing method to produce a capsule.

2-4. Preparation of gel

ingredient weight% Tofu extract 0.01 Butylene glycol 2.00 glycerin 1.00 Disodium EDTA 0.02 PEG-60 Hydrogenated Castor Oil 1.00 Cocamidopropyl Betaine 4.00 Lauryl Glucoside 6.00 Polysorbate 20 1.00 Sodium benzoate 0.40 Caprylic Glycol 0.20 Sodium lactate 0.50 Hydroxyethyl cellulose 0.7 Purified water 83.17 aggregate 100.0

2-5. Preparation of ointment

ingredient weight% Tofu extract 0.01 diethanolamine 1.5 Polyvinyl photolidone 5.0 Propylene glycol 30.0 Purified water 63.49 aggregate 100.0

Claims (7)

A cosmetic composition for suppressing or improving white hair, vitiligo, or hypopigmentation, containing Sargassum fusiforme extract as an active ingredient.
A cosmetic composition characterized in that the above-mentioned tangle extract is manufactured by a method including: (S10) a step of first extracting tangle with hot water; (S20) a step of removing alginic acid from the above-mentioned tangle first extract and drying it; (S30) a step of secondly extracting the above-mentioned tangle first extract from which the alginic acid has been removed and dried by precipitating it in an organic solvent; and (S40) a step of obtaining a supernatant of the above-mentioned tangle second extract. In claim 1, the cosmetic composition has a formulation selected from the group consisting of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray. delete A cosmetic composition for hair care that promotes melanin production in hair and suppresses gray hair by applying the cosmetic composition according to Article 1 to the hair. A pharmaceutical composition for the prevention or treatment of white hair, vitiligo, or hypopigmentation, containing an extract of Sargassum fusiforme as an active ingredient.
A pharmaceutical composition characterized in that the above-mentioned tangle extract is manufactured by a method including: (S10) a step of first extracting tangle with hot water; (S20) a step of removing alginic acid from the above-mentioned tangle first extract and drying it; (S30) a step of secondly extracting the above-mentioned tangle first extract from which the alginic acid has been removed and dried by precipitating it in an organic solvent; and (S40) a step of obtaining a supernatant of the above-mentioned tangle second extract. A composition for external use in the skin for the prevention or treatment of white hair, vitiligo, or hypopigmentation, containing an extract of Sargassum fusiforme as an active ingredient,
A skin external preparation composition characterized in that the above-mentioned tangle extract is manufactured by a method including: (S10) a step of first extracting tangle with hot water; (S20) a step of removing alginic acid from the above-mentioned tangle first extract and drying it; (S30) a step of secondly extracting the above-mentioned tangle first extract from which the alginic acid has been removed and dried by precipitating it in an organic solvent; and (S40) a step of obtaining a supernatant of the above-mentioned tangle second extract. A food composition for preventing or improving white hair, vitiligo, or hypopigmentation, containing an extract of Sargassum fusiforme as an effective ingredient.
A food composition characterized in that the above-mentioned tangle extract is manufactured by a method including: (S10) a step of first extracting tangle with hot water; (S20) a step of removing alginic acid from the above-mentioned tangle first extract and drying it; (S30) a step of secondly extracting the above-mentioned tangle first extract from which the alginic acid has been removed and dried by precipitating it in an organic solvent; and (S40) a step of obtaining a supernatant of the above-mentioned tangle second extract.