WO2020067663A1 - Composition for promoting skin turnover and melanin excretion - Google Patents

Abstract

The present invention relates to a cosmetic composition, a pharmaceutical composition, a quasi-drug composition, and a health functional food, each of which comprises vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient for skin regeneration or skin whitening. According to the present invention, differentiation in the stratum basale under the epidermis is activated to shorten the skin turnover period, thereby rapidly exfoliating old calluses, promoting skin regeneration, reducing pigmentation in the skin, inhibiting melanin synthesis, and promoting melanin excretion ex vivo, with the resultant effect of skin whitening and hair growth.

Description

Composition for promoting skin turnover and promoting melanin discharge

The present invention comprises vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient for skin regeneration by promoting skin turnover or skin whitening cosmetic composition by promoting melanin discharge, pharmaceutical composition, quasi-drug composition and It is about health functional food.

The skin is divided into three parts: the upper skin layer, the epidermal, dermal and subcutaneous fat layers. Among them, the epidermis is composed of a base layer, a polar layer, a granular layer, and an outermost layer of the stratum corneum, and the epidermal cells of the base layer are differentiated as they rise to the top of the skin, and finally reach the stratum corneum. The epidermal cells that have reached the stratum corneum are lost to the nucleus and filled with an insoluble protein called keratin and converted into dead cells. The stratum corneum is composed of differentiated epidermal cells (keratinocytes) and skin lipids that fill in between them. It prevents substances in the body from going out and protects the human body from external physical, chemical, and biological stimuli. do.

If such a stratum corneum does not peel off normally and remains on the skin surface, the stratum corneum thickens and the face color becomes dull and dark. The impurities remaining on the skin surface or hair follicles are oxidized or decomposed by oxygen or microorganisms, and these substances cause skin troubles such as inflammation. In addition, aging skin, dry skin, acne skin, etc., the separation of the stratum corneum layer becomes slower than normal, and the stratum corneum thickens (stratum corneum stratification), and a remarkable feature that appears on the skin appearance is the occurrence of skin scales. Keratin layering is mainly due to reduced moisturizing ability of the skin, desmosome (protein that connects keratinocytes) degrading enzymes and reduced activity, decreased cell activity, and causes skin aging, UV exposure, and pollution.

By artificially thinning the stratum corneum thickened by these internal and external factors, the activity or regeneration of living cells under the stratum corneum increases, reducing skin scales appearing on the skin's appearance and softening the skin, removing wrinkles, acne There are many studies to solve keratin stratification within a range in which irritation is not induced due to effects such as occurrence inhibition and treatment.

Specifically, in order to eliminate keratin layering, the skin is physically rubbed or chemical dermabrasion is used. In particular, chemical peeling has additional effects such as improving fine lines, relieving rough skin, and removing blemishes. As for the concentration of alpha hydroxy acid used for chemical peeling, a high concentration of at least 20-30% or more is used, but at a low concentration of less than 2/4 10%, the stratum corneum is gradually peeled off (J.AM.Acad. Dermatol., 11, 867 -879, 1984; Van Scott, etc.), skin moisturizing enhancement (happi, jully, 66-68, 1994; Tom, etc.), fine wrinkle relief (Cutis, 43, 222-228, 1989; Van Scott, etc.) Recently, many products have been released in the cosmetics and pharmaceutical industries. However, since alpha hydroxy acid has disadvantages that cause skin irritation and irritation when the pH is low, it is very important to adjust the concentration and pH of the skin to alleviate skin side effects. In addition, it is possible to reduce the side effects of skin by attaching proteins, lipids, etc. to alpha hydroxy acid, or by adding a side effect reliever and changing the structure, but the effects such as exfoliation and improvement of fine lines are also reduced, thereby minimizing side effects and maximum effects. There is a problem that can not be obtained.

In addition, sensitive skin refers to skin that is more sensitive to external irritants, allergic substances or environmental changes or internal causes of the human body, which are more sensitive to skin irritation or dermatitis, and protects sensitive skin from skin irritants. It has been argued that a healthy stratum corneum is the most important factor in doing so. In fact, since the degree of skin irritation tendency and skin permeability are correlated, skin barrier failure is also a cause of sensitive skin. Therefore, strengthening the skin barrier can also be one of the ways to normalize sensitive skin. So far, cosmetic products for sensitive skin have been developed so that they do not cause contact dermatitis, allergic dermatitis, etc. by removing active ingredients that may cause irritation. However, due to environmental and psychological factors, the use of active ingredients is also required in cosmetics of sensitive skin holders. Therefore, there is a need to develop a cosmetic having a function of enhancing a skin barrier and promoting a skin turnover cycle to improve and normalize sensitive skin.

On the other hand, with the increase in leisure population, the number of people enjoying outside activities is getting worse, and melanin pigmentation by UV rays is getting worse, and hyperpigmentation is a serious mental burden from the skin beauty point of view, which can interfere with normal social activities. do. Melanin, which determines human skin color, is produced by melanocytes, and enzymes such as tyrosinase exist in the melanocytes, and they work together to form an amino acid called tyrosine, which is always present in the body. Through the oxidation reaction of polymerization, melanin, a dark brown pigment, is formed. The melanin thus formed moves to the epidermal cells called keratinocytes through the dendritic projections of melanocytes. Here, melanin forms a hat-like structure around the nucleus, which plays an important role in protecting genes from ultraviolet rays and protecting proteins in cells by removing free radicals. However, when more melanin is produced than necessary, hyperpigmentation such as freckles, freckles, and spots is induced, which results in a bad cosmetic effect. Women in the Asian region have preferred white and fine skin for a long time, and taking this as an important criterion for beauty, the desire for prevention and improvement of skin pigment abnormalities and hyperpigmentation is increasing, thereby preventing the overproduction of melanin. The development of whitening cosmetics and drugs for the purpose is actively underway.

However, there is no enzyme that breaks down melanin in the living body, but it is removed by exfoliating off the skin when keratin comes off the epidermis. Therefore, by promoting such a turn-over cycle, it is necessary to search for a substance capable of quickly discharging melanin produced in the skin epidermis to the body and using it to expel melanin to the body to increase the whitening effect. .

Fine wrinkles and deep wrinkles are also one of the major causes of uneven skin texture, and retinoic acid, a cosmetic ingredient for wrinkle improvement, is most frequently prescribed, but strong skin irritation caused by retinoic acid is a big problem to be solved.

On the other hand, the hair has a constant hair cycle (Hair cycle), keratinocytes grow through the growth phase, which is the period during which the epidermal cells (keratinocyte) around the breast nipple divide and proliferate, and the degeneration phase, which is the time when hair growth stops and separates from the hair nipple. They do not do it, and then they go through a rest period that falls out after a certain period of time. The parent cycle repeats this process of growth, degeneration, rest, and growth. Baldness is not a phenomenon in which the hair does not fall out, but gradually becomes thinner and fluffy, and as the baldness progresses, the papillae existing in the hair follicle become smaller. As the nipple becomes smaller, the hair cycle becomes shorter and the newly grown hair becomes thinner. At the beginning of hair loss, it is known that the use of the topical coating agent minoxidil and the oral drug propecia can slow the progression to a certain extent, but it is difficult to restore normal hair loss to the normal state by lengthening the hair cycle.

Currently, several studies have been conducted on the relationship between skin turnover and hair growth, and for example, it has been reported that deficiency of vitamin D receptor is inhibited not only in epidermal turnover but also in growth of hair follicles (J. Invest. Dermatol., 118 (1)) , 11-6, 2002; Xie Z et al.). Since proliferation of keratinocytes is essential for both the skin turnover and hair growth stages, the effect of promoting skin turnover may exhibit hair loss prevention or hair growth efficacy.

The inventors of the present invention have completed the present invention to solve the above problems, and the composition of the present invention activates differentiation in the base layer in the epidermis to accelerate the turnover cycle of the skin, thereby rapidly removing old keratin to promote skin regeneration, , It improved the hair cycle and reduced the pigmentation in the skin, as well as the effect of less irritation to the skin.

One object of the present invention is to provide a cosmetic composition for skin regeneration or skin whitening comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for skin regeneration or skin whitening comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a quasi-drug composition for skin regeneration or skin whitening comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a health functional food for skin regeneration or skin whitening comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.

The composition of the present invention penetrates into the base layer in the epidermis and induces differentiation directly, thereby activating differentiation of epidermal cells to accelerate the turnover cycle of the skin to quickly remove old keratin to promote skin regeneration and to secrete melanin pigment By suppressing melanin secretion from the skin, it reduces the pigmentation in the skin and promotes the extracellular release of melanin, which has a skin whitening effect, but also has less irritation to the skin and can promote hair growth.

1 is a result of screening a single compound having trypsin resistance.

2 is a result showing mitochondria lost by menadione.

3 is a result showing a nucleus that is not degraded by menadione.

4 is a result showing the differentiation marker changes in keratinocytes by menadione.

5 is a result showing the increase in the thickness of the artificial skin by menadione.

6 is a result showing the keratin turnover by menanion.

7 is a result of analyzing the amount of biosynthesized melanin in the artificial epidermis by treatment with 0.1% menadione.

8 is a result showing the distribution of melanin in the artificial epidermis by treatment with 0.1% menadione.

9 is a result showing the hair growth of pigs and other skin caused by Medanion.

Specifically, it is as follows. Meanwhile, each description and embodiment disclosed in the present invention can be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. In addition, the scope of the present invention is not limited by the specific descriptions described below.

In the present invention, vitamin K3 (menadione) activates differentiation in the basement layer in the epidermis to accelerate the turnover cycle of the skin, thereby quickly removing old keratin, promoting skin regeneration, reducing pigmentation in the skin, and irritating Little effect was confirmed, and the present invention is based on this.

Hereinafter, the configuration of the present invention will be described in detail.

The first aspect of the present invention is to provide a cosmetic composition for skin regeneration or skin whitening comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.

The term "skin" as used in the present invention refers to a tissue that covers the body surface of an animal, and is the broadest concept including scalp and hair, as well as tissue that covers a body surface such as a face or body. Meanwhile, in the present specification, the skin may include not only biological skin, but also artificial skin or a skin mimetic that embodies the state of biological skin.

The term “turn-over” used in the present invention means a process in which new cells made in the dermal layer rise to the stratum corneum and become dead cells, and again, new cells are born in the base layer, depending on the site or age. Although different, the turnover period of the epidermis in normal skin can be 4 to 6 weeks. The skin turnover promotion is achieved by promoting differentiation of epidermal cells, and is characterized by promoting skin turnover by penetrating the base layer and inducing direct differentiation. In addition, the skin turnover promotion can be achieved by increasing the thickness of the epidermal layer of the skin. Skin regeneration may be promoted by promoting the skin turnover.

The term “skin regeneration” used in the present invention refers to a process of restoring skin tissue to damage caused by external and internal causes of skin, and increases skin moisturization, promotes skin elasticity, increases skin thickness, improves skin wrinkles, and relieves skin irritation And skin damage recovery actions, but are not limited thereto. In the present invention, the differentiation of epidermal cells is promoted due to the promotion of skin turnover, and ultimately, it has a skin regeneration effect.

The term "epidermis" as used in the present invention is a tissue that is always in contact with the external environment and mainly protects the human body from external physical damage and chemicals, and prevents bacteria, fungi, viruses, etc. from invading the skin. It means that it acts as a protective barrier against moisture loss.

The term "differentiation of epidermal cells" as used in the present invention means a process in which keratinocytes functionally differentiate from the base layer, the lowest layer of the skin, to gradually form a polar layer, a granular layer, and a stratum corneum. The composition of the present invention is characterized by promoting the differentiation of epidermal cells by penetrating the base layer and inducing direct differentiation. In addition, acceleration of differentiation of the epidermal cells can be achieved by increasing the thickness of the epidermal layer of the skin.

The "keratinocytes" are the main constituent cells of the epidermis, and after division, they move upwards, and then, differentiation begins to take place, and through the final differentiation process, they play a role in creating a skin barrier through division and differentiation. When the final differentiation of keratinocytes into the stratum corneum occurs, all the organelles such as the nucleus and mitochondria in the cell are decomposed.

The term "keratinization" or "keratinization" as used in the present invention means that the stratum corneum is formed as the last step of differentiation of the epidermis.

The term “stratum corneum” or “stratum corneum” used in the present invention is a main component that is distributed in the skin, hair, and nails and constitutes a cell skeleton, and differentiated epidermal cells (keratinocytes) and skin lipids that fill in between them. ), Which means that it prevents substances from going out to the body and protects the human body from external physical, chemical, and biological stimuli. The keratinocytes are continuously made inside, but the old keratin of the outermost layer is peeled off, so it maintains a constant thickness. The epidermal cells that have reached the stratum corneum are transformed into dead cells as the nucleus is lost and filled with the insoluble protein keratin.

The term “stratum corneum layering” used in the present invention means a phenomenon in which the stratum corneum thickens because the stratum corneum layer is not normally peeled off and remains on the skin surface. The composition of the present invention may improve skin stratum corneum by promoting skin turnover with less side effects of skin irritation.

The term “keratin” used in the present invention is an intermediate filament constituting the cytoskeleton of keratinocytes, and refers to a characteristic protein component produced in the process of differentiation of keratinocytes. . To date, 54 types of keratin genes have been found and are divided into type I and type II keratin according to protein size (Kim, 2009). The two form a pair. In the keratinocytes of the base layer, K5 / K14 keratin, a type of keratin, is expressed, and when cell differentiation begins in the polar layer, their expression is suppressed and K1 / K10 keratin pairs begin to appear (Kwon et al. , 2009).

As used in the present invention, the term "melanin (melanin)" is to determine the skin color of a person, which is produced in melanocytes (melanocyte), which enzymes such as tyrosinase exist, and they work together to always exist in the body Through the oxidation reaction of polymerizing an amino acid called tyrosine into a substrate, melanin, a dark brown pigment, is formed. The melanin thus formed moves to the epidermal cells called keratinocytes through the dendritic projections of melanocytes. Here, melanin forms a hat-like structure around the nucleus, which plays an important role in protecting genes from ultraviolet rays and protecting proteins in cells by removing free radicals. However, if more melanin is produced than necessary, hyperpigmentation such as freckles, freckles, and spots may occur.

As used in the present invention, the term "hyperpigmentation" refers to the phenomenon of hyperpigmentation caused by increased melanin in the skin, nails, or mucous membranes surrounding the oral cavity or nasal cavity.

By promoting the skin turnover cycle through the present invention, it is possible to improve the whitening effect by promoting the extracorporeal discharge of melanin produced in the skin epidermis.

The composition of the present invention can suppress melanin secretion, thereby alleviating or improving melanin deposition or hyperpigmentation, and improving the whitening effect.

The "whitening" means to whiten the skin, and it means to alleviate or improve various pigmentation including freckles and freckles caused by excessive synthesis of melanin or melanin deposition or hyperpigmentation. The composition of the present invention can accelerate the skin turnover cycle to promote the extracorporeal release of melanin, inhibit melanin secretion, and alleviate or improve melanin deposition or hyperpigmentation.

The term "improvement" used in the present invention activates differentiation of epidermal cells that are reduced by external stimulation or aging, thereby accelerating skin turnover cycles, rapidly removing old keratin, promoting skin regeneration, and secreting melanin pigment. It may include all the actions to reduce the pigmentation in the skin by inhibiting melanin secretion in the artificial skin, but is not limited thereto.

The term "vitamin K" used in the present invention is a fat-soluble 타 vitamin, which is well known as a fat-soluble compound present in the leaves of plants, also called Menadione, and vitamin K3 can be made synthetically.

The composition in the present invention, the vitamin K3 (menadione) may contain a content of 0.01 to 1% based on the total weight of the cosmetic composition, specifically may contain a content of 0.05 to 0.7%, more specifically As may include a content of 0.1 to 0.5%, but is not limited thereto,

The composition in the present invention is resistant to trypsin.

The term "pharmaceutically acceptable salt" used in the present invention means a salt in a form that can be used pharmacologically among salts in which cations and anions are bound by electrostatic attraction, and is relatively non-toxic to patients. As a concentration having a harmless effective effect, it means any organic or inorganic addition salt of the above compound which does not lower the beneficial effect of the present invention due to side effects caused by this salt.

The composition of the present invention was found by screening for a substance that has resistance to trypsin in keratinocytes and causes mitochondria to disappear by causing differentiation. Through the epidermal thickness measurement method of artificial skin, it was confirmed that the effect of promoting the differentiation of epidermis in the case where 0.1%, 0.5% of menadione was applied on artificial keratin and 0.5% of tretinoin was applied. In addition, when the above combination was applied to the skin turnover enhancement effect evaluation method by the DHA staining method, it was confirmed that it has superior keratin turnover enhancement efficiency compared to the uncoated group. This showed similar turnover efficacy in live pig skin, and it was confirmed that this effect of promoting epidermal differentiation was not a mechanism for promoting turnover by exfoliation.

The composition of the present invention is characterized by inhibiting melanin secretion in artificial skin secreting melanin pigment. When the degree of skin irritation when promoting epidermal differentiation and suppressing melanin secretion by menadione treatment was confirmed by the secretion amounts of the inflammatory markers IL-1α and IL-8, menadione was found to have a lower secretion of the inflammatory markers than tretinoin. In addition, it was confirmed through the patch test on the human skin that there was no irritation to the skin.

The menadione of the present invention is characterized by promoting hair growth. The hair has a constant hair cycle, and keratinocytes do not grow through the growth phase, which is the period during which the keratinocytes around the breast nipple divide and proliferate, and the degeneration phase, when the hair stops growing and separates from the hair nipple. After a certain period of time, it goes through a resting period. The composition of the present invention is characterized by exhibiting a hair loss prevention or hair growth effect as well as promoting skin turnover around the nipple.

In the present invention, it was confirmed that hair growth is promoted when menadione is treated on the back skin of a pig. Through this, it was confirmed that menadione has a skin turnover effect similar to tretinoin and suppresses melanin synthesis and simultaneously exhibits low skin irritation level and promotes skin turnover to exert hair growth efficacy.

The term "cosmetic composition" used in the present invention is a solution, external ointment, cream, foam, nutrition lotion, soft lotion, pack, softener, emulsion, makeup base, essence, soap, liquid detergent, bathing agent, sunscreen cream, sun It can be prepared in a formulation selected from the group consisting of oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays. , But is not limited thereto. Specifically, the cosmetic composition of the present invention may be preferably prepared as a semi-solid preparation such as ointment for external use, lotion, etc., but is not limited thereto.

In addition, the cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers to be blended with general skin cosmetics, and for example, oil, water, surfactants, moisturizers, and lower alcohols as common ingredients. , Thickeners, chelating agents, pigments, preservatives, fragrances, etc. may be appropriately blended, but is not limited thereto. The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation.

When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. as carrier components This may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component, and in the case of a spray, additionally, chlorofluorohard Propellants such as locarbon, propane / butane or dimethyl ether, but are not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butyl glycol oil, and the like can be used, and in particular, cottonseed oil, peanut oil, corn seed oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan May be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, micro Crystalline cellulose, aluminum metahydroxide, bentonite, agar or trakant may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a soap, alkali metal salt of fatty acid, fatty acid hemiester salt, fatty acid protein hydrolyzate, isethionate, lanolin derivative, aliphatic alcohol, vegetable oil, glycerol, sugar, etc. are used as carrier components. It can be, but is not limited to. These may be used alone or in combination of two or more.

The vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof of the present invention may be contained in an amount of 0.0001% to 10% by weight, more specifically 0.0005% by weight, based on the total weight of the total cosmetic composition. % To 10% by weight, but is not limited thereto.

The cosmetic composition of the present invention is a solution, ointment for external use, cream, foam, nutrient lotion, soft lotion, pack, soft water, emulsion, makeup base, essence, soap, liquid detergent, bathing agent, sunscreen cream, sun oil, suspension, Emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches, and sprays may be those having a formulation selected from the group consisting of, but not limited to It is not.

The second aspect of the present invention provides a pharmaceutical composition for skin regeneration or skin whitening comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.

The term "pharmaceutical composition" used in the present invention, in addition to including vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient, a suitable carrier, excipient and commonly used in the manufacture of a pharmaceutical composition Diluents may be further included.

At this time, "pharmaceutically acceptable salt" of the present invention means a salt in a form that can be used pharmaceutically among salts, which are materials in which cations and anions are bound by electrostatic attraction, and are usually metal salts or organic bases. It may be a salt with, a salt with an inorganic acid, a salt with an organic acid, a salt with a basic or acidic amino acid, but is not limited thereto.

Specifically, the pharmaceutical composition may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, and sterile injection solutions, respectively, according to a conventional method. Examples of carriers, excipients, and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, and methyl Cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but are not limited thereto. These may be used alone or in combination of two or more.

When formulating the pharmaceutical composition of the present invention, it may be prepared by using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are usually used.

Solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc. These solid preparations include one or more excipients such as starch, calcium carbonate, sucrose, lactose, or gelatin. It can be prepared by mixing. In addition, lubricants such as magnesium stearate and talc may be used in addition to simple excipients. Liquid preparations for oral administration include suspending agents, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used as diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be included. have.

Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, external preparations, and the like. As the non-aqueous solvent and suspension, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used.

The pharmaceutical composition of the present invention may be used as an external preparation for skin and applied to the skin. The pharmaceutical composition may be a formulation of a liquid, suspension, emulsion, cream, hydrogel, lotion, ointment, spray, patch, or aerosol, but is not limited thereto. When used as an external preparation for skin, fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic Emulsifiers, fillers, metal ion blockers and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in external preparations for the skin It may contain adjuvants commonly used in the field of dermatology.

The pharmaceutically effective amount and effective dose of the pharmaceutical composition of the present invention may be varied by a method of formulating the pharmaceutical composition, a method of administration, an administration time and / or a route of administration, and the like to achieve the administration of the pharmaceutical composition. The type and extent of the reaction to be performed, the type of the subject to be administered, the age, weight, general health condition, the severity or severity of the disease, sex, diet, excretion, components of the drug or other composition used simultaneously or simultaneously with the subject It can be varied according to various factors including the like and similar factors well known in the pharmaceutical field, and a person skilled in the art can easily determine and prescribe an effective dosage for the desired treatment.

Administration of the pharmaceutical composition of the present invention may be administered once a day, it may be divided into several times. Therefore, the above dosage does not limit the scope of the present invention in any way.

The route of administration and the mode of administration of the pharmaceutical composition of the present invention may be independent of each other, and are not particularly limited in the manner, and any route of administration and administration as long as the pharmaceutical composition can reach the target site of interest You can follow the way. The pharmaceutical composition may be administered by oral administration or parenteral administration, preferably parenteral administration.

As the method of parenteral administration, for example, intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration, or subcutaneous administration may be used, and the composition may be applied to a diseased site, sprayed, or inhaled. May, but is not limited to these.

The pharmaceutical composition of the present invention may be administered by topical administration, more preferably by transdermal administration during parenteral administration, more preferably by applying the pharmaceutical composition to the skin of an individual.

The term "individual" used in the present invention means all animals, including, but not limited to, mammals including rats, livestock, humans, and the like. In addition, in the present invention, the term "applying" means contacting the composition of the present invention to the skin of an individual in any suitable way, thereby including all actions aimed at absorbing the composition into the skin. , But is not limited to this.

The third aspect of the present invention is to provide a quasi-drug composition for skin regeneration or skin whitening comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.

The term "quasi-drug composition" used in the present invention, in addition to containing at least one compound selected from the group consisting of hydroxycinnamic acid, isoamyl acetate and betaine or a pharmaceutically acceptable salt thereof as an active ingredient, if necessary It may further include a pharmaceutically acceptable carrier, excipient or diluent. The pharmaceutically acceptable carrier, excipient or diluent is not limited as long as it does not impair the effects of the present invention, for example, fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, lubricants, sweeteners, fragrances, preservatives, etc. It can contain.

The quasi-drug composition of the present invention may be exemplified by disinfecting cleansers, shower foams, ointments, wipes, coatings, and the like, and may preferably be made of semi-solid preparations such as ointments for external use, lotions, etc., but is not limited thereto. The formulation method, dosage, method of use, ingredients, etc. of the quasi-drug can be appropriately selected from conventional techniques known in the art.

The fourth aspect of the present invention is to provide a health functional food for skin regeneration or skin whitening comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.

The term "health functional food" used in the present invention is the same term as a specific health food, a functional food, and a health food, and refers to a food with high medical effect and medical effect processed to efficiently exhibit bioregulatory functions in addition to nutrition. , The food may be prepared in various forms such as tablets, capsules, powders, granules, liquids, pills, etc. in order to obtain useful effects in preventing or improving hair loss-related diseases. Unlike general medicines, it has the advantage that there is no side effect, etc. that can occur when taking medicines for a long time by using food as a raw material, and it has excellent portability, and the health functional food of the present invention can be ingested as a supplement for preventing or improving diabetes. .

There is no limitation in the form that the health functional food of the present invention can take, it may include all foods in a common sense, and it is interchangeable with terms known in the art, such as functional food. Specifically, the food is a food prepared by adding the food composition of the present invention to food materials such as beverages, teas, spices, gums, confectionery, or by encapsulation, powdering, suspension, etc., for example, various foods, beverages , Gum, tea, vitamin complex, health functional foods, etc. can be used. It may be prepared by mixing a known additive with other suitable auxiliary ingredients that may be included in food according to the choice of those skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gums, ice cream, various soups, beverages, teas, drinks, alcoholic beverages, and There are vitamin complexes, etc., and can be prepared by adding the compound represented by Chemical Formula 1 according to the present invention as a main component to juice, tea, jelly, and juice. It also includes foods used as feed for animals.

Hereinafter, examples and the like will be described in detail to help understanding of the present invention. However, the embodiments according to the present invention can be modified in many different forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

Example 1: Trypsin resistance evaluation

Extracellular matrix (Extracellular matrix) is a complex of proteins, glycoproteins, polysaccharides, etc. that form the microenvironment outside the cell, as well as the physical support of the cell, as well as the signal transmission between the cell and the cell, and the cell's transport pathway It also regulates differentiation. Hyaluronic acid (hyaluronic acid) is a chain-like polymer polysaccharide in which glucuronic acid and N-acetylglucosamine residues are repeatedly connected.It is a major component of the extracellular matrix in the epidermis, which promotes biosynthesis of hyaluronic acid when epidermal differentiation occurs (Journal of Investigative Deramatology (2014) 134, 2174-2182, tetrahydrojamoic acid). Accordingly, in the present invention, a single compound library held in the company was treated with human keratinocytes, respectively, and a substance that was resistant to trypsin that breaks down the extracellular matrix was screened.

Specifically, skin keratinocytes were inoculated in a 24-well microplate at a concentration of 2 x 10 ⁴ cells / well, and cultured for 24 hours in a CO ₂ incubator at 37 ° C and 5% concentration. Then, the medium was removed, and a single compound was added to the culture medium (DMEM) at 5 ppm each, and treated with cells to incubate for 48 hours. Then, the plate was washed twice with PBS, and 0.05% trypsin was added to each well, followed by incubation for 7 minutes in a CO ₂ incubator. Once again, the cells supported by PBS are removed, and then the culture medium is added and cultured. Cells counting kit-8 (DOJINDO) CCK-8 (Cell Counting Kit-8, DOJINDO) is used to define cells remaining on the plate in addition to the cells that have been degraded by extracellular matrix by trypsin as cells resistant to trypsin and to measure the number of cells attached to the plate. Laboratories) was performed as follows. After adding CCK-8 solution to 10% of 10 culture medium, incubating for 1 hour in a CO ₂ incubator at a concentration of 37 ° C., 5%, and absorbance at 450 nm was measured using a spectrophotometer. Trypsin resistant substances Caffeic acid, Menadione, Rosmarinic acid, Gallic acid were found (Fig. 1).

Example 2: Staining of mitochondria and nuclei

Acta Histoche. Cytochem. 32 (6) 465-476, 1999 reported that during the epidermal differentiation process, mitochondria and nuclei were absorbed and degraded by autophagosomes. It was confirmed that the mitochondria and nuclei in the cells were decomposed while differentiating into keratinocytes by the trypsin-resistant menadione discovered above.

The keratinocytes were incubated for 48 hours after treatment with 1 ppm and 5 ppm of menadione. Subsequently, mitotracker or DAPI (4`, 6-diamidino-2-phenylindole) fluorescent dye was added to the culture medium to stain the mitochondria and nuclei, followed by fluorescence microscopy. In FIG. 2, it was observed that mitochondria disappeared by menadione, and it was found that differentiation was occurring. In FIG. 3, it was found that the nucleus was not decomposed by menadione, and it was found that the terminal differentiation was not performed due to keratin.

Example 3: In vitro cell differentiation measurement

After inoculating HaCaT cells into a 6-well plate, the next day, 5 ppm of menadione was treated, and then the cells were eluted with 5X lysis buffer to obtain proteins. Cell debris were removed by centrifugation and quantified using a BCA protein assay kit (Pierce, USE). Cell extract proteins were separated using 4-15% SDS-PAGE gel (Bio-Rad, USA) and transferred to a nitrocellulose membrane (Life Technologies, New Zealand). After blocking Membrane with TBST buffer containing 3% BSA for 1 hour, it was reacted for 1 day at 4 ° C in a buffer solution containing involucrin antibody expressed in the stratum granulosum and K14 antibody expressed in the stratum basale. . After washing the Membrane with TBST and reacting with a secondary antibody bound with peroxidase, color was developed using enhance chemiluminescence (Amersham, UK), and the immune-reactive band was confirmed with Fusion FX 5 image system (Vilber Lourmat, France). . As can be seen in FIG. 4, when 5 ppm of menadione was treated, it was found that the amount of Keratin 14 was decreased and the expression of involucrin was increased, indicating that differentiation in cells was promoted.

Example 4: Measurement of artificial skin epidermal thickness

A 3D skin model was used to confirm that the intraepidermal differentiation was actually promoted by menadione. The reconstructed human epidermis model (Neoderm-E, Tego Science, Korea) was transferred to a 6-well plate, stabilized in 2 mL of growth media (Tego Science, Korea) for 24 hours, and menadione in triethylhexanoin (TIO) 0.1%, 0.5%, negative control TIO, and tretinoin 0.01% as a positive control were applied on the keratin. After 5 days, the artificial skin tissue was stained with H & E to check whether the epidermal thickness increased.

As a result, as can be seen from Table 1 and FIG. 5, it was confirmed that the epidermal thickness increased by 36% compared to the control by 0.1% treatment with menadione. The positive control tretinoin (Tretinoin) was treated with 0.01%, showing a 43% increase in efficacy compared to tretinoin, but it was found that keratin differentiation is promoted by menadione. At this time, as a result of confirming the discharge of IL-1α and IL-8, stimulation markers secreted from the artificial skin, it was found that the stimulation level was significantly lower than tretinoin. Table 1 below shows the increase in epidermal thickness and the degree of irritation caused by menadione.

	Epidermis Thickness (νm)	IL-1α (pg / ml)	IL-8 (pg / ml)
Control	58 ± 5.97	133.7	107
Tretinoin 0.01%	83.5 ± 2.66	269.9	72
Menadione 0.1%	79 ± 4.72	160.3	169

Example 5: Pig skin epidermal thickness measurement and exfoliation evaluation

Exp Dermatol. According to 2012 Sep; 21 (9): 643-9, physical stimulation of the stratum corneum (peeling, tensile force, etc.) can promote epidermal differentiation, and pig skin is examined to see if the effect of promoting epidermal differentiation by menadione is due to exfoliation. Using it, menadion's epidermal differentiation efficacy and peeling efficacy were measured. Specifically, a 1 mm thick back skin of a slaughtered pig was sampled in a 24-well plate size, washed twice with PBS and betadine, placed on a cell culture insert, and added to the plate with culture medium (DMEM) to add 37 ° C. , Incubated for 3 days in a 5% concentration CO ₂ incubator. At this time, 100 ppm of menadione was treated in one culture medium so that pig skin could be absorbed. As can be seen in Figure 5, it was found that the epidermal thickness is increased by menadione even in pig skin.

For the evaluation of exfoliation, the exfoliation evaluation method mentioned in the Journal of the European Academy of Dermatology and Venereology, 2014, 28, 415-423 was used as a modification. In detail, a 1 mm thick pig skin was taken with a biopsy tool and a 6 mm diameter sample was placed in a 96-well plate, washed with PBS once, and 100 ul of the sample to be tested was added. This sample is plotted with a sample containing only solvent (DW / DMI) as a negative control and 5% serine pH 6 as a positive control by measuring the number of dead keratin cells after storage for one day at 37 ° C and 50% humidity. Did. The results were compared with the results of menadione 0.1% and 0.5%, and it was confirmed that there was no keratin peeling effect by menadione concentration. As a result, it was found that the effect of promoting mendione epidermal differentiation is not a secondary differentiation effect due to exfoliation of the keratin, but is that it penetrates into the base layer and induces differentiation directly.

Example 6: Evaluation of keratin turnover enhancement by DHA staining

Subjects were enrolled in 14 healthy men and women between the ages of 20 and 40. Before applying the sample, the color of the lower arm and upper arm of the individual was measured with a chroma meter, and then about 0.4 ml of dihydroxy acetone (DHA) at a concentration of 10% was placed inside the upper and lower arm of the arm for 6 hours. While attached. After 24 hours, the color of the brown-colored area was measured by DHA to compare the color difference from before the sample was applied. Thereafter, while applying the sample once a day, the degree of discoloration every day was measured with a chromaometer, and the time taken to return to the original skin color was measured. In this experiment, the time taken for the DHA-colored skin to return to its original state was indicated through a relative comparison analysis using a regression analysis method.

When the result of the untreated group was calculated to be 9 days, when the menadione and caffeic acid found in the trypsin resistance experiment were tested, when 0.1% of menadione was used, the degree of return of colored skin compared to untreated was about 1.3 days. It was pulled, and in the case of 0.5% of menadione, it was found that it was pulled forward about 2 days. All combination groups did not cause side effects such as erythema, itching and burning sensation during and after the experiment. Through this, it was found that menadione, which is a substance that promotes epidermal differentiation in artificial skin and pig skin, actually promotes turnover in human skin (FIG. 6, Table 2). Table 2 below shows the color change of dyed keratin caused by menadione.

Substance name	ΔL * (L * Day9-L * Day0)	p -value (vs TIO)
TIO	2.948	-
Vitamin K3 0.1% / TIO	3.453	0.054322
Vitamin K3 0.5% / TIO	3.752	0.05015

Example 7: Artificial skin melanin inhibition

In order to confirm that the biosynthesis of melanin in the epidermis is reduced by menadione, a 3D skin model including a melanocyte was used. Reconstructed human pigmented epidermis model (Neoderm-ME, Tego Science, Korea) was transferred to a 6-well plate, stabilized in 2 mL of growth media (Tego Science, Korea) for 24 hours, and menadione in triethylhexanoin (TIO) 0.1%. With and without treatment, Hydroquinone (HQ) 4% and Niacinamide (NA) 2% were applied on the keratin as a positive control. After 5 days, the artificial skin tissue was stained with Fontana-Masson to check the distribution of melanin in the epidermis.

As a result, as can be seen from Table 3 and Figures 7 and 8, as a result of analyzing the amount of melanin biosynthesized in the epidermis by treatment with 0.1% menadione, it was confirmed that the decrease was 63.87% compared to the control group. The positive control of Hydroquinone (HQ) 4% and Niacinamide (NA) 2% was 75.92% and 59.63% compared to Control, and Menadione (MD) 0.1% treatment was lower than Hydroquinone 4% but higher than Niacinamide 2%. I could confirm.

Substance name	Melanin contents (% of control)	Stdev
Control	100.00	37.53
Hydroquinone	24.08	12.03
Niacinamide	40.37	16.55
Menadione	36.13	17.04

Example 8: Pig skin hair dye

Pig skin was cultured and H & E stained as in the experiment for measuring epidermal thickness using pig skin of Example 5 above. It was confirmed that hair was newly grown on the menadion-treated skin (FIG. 9).

From the above description, those skilled in the art to which the present invention pertains will appreciate that the present invention may be implemented in other specific forms without changing its technical spirit or essential features. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the following claims rather than the detailed description and equivalent concepts thereof.

Claims (12)

1. Vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient, for skin regeneration or skin whitening cosmetic composition.
2. The cosmetic composition according to claim 1, wherein the skin regeneration is achieved by promoting differentiation of epidermal cells.
3. The cosmetic composition according to claim 1, wherein the skin regeneration is performed by penetrating the base layer and inducing direct differentiation.
4. According to claim 1, The skin regeneration is made by increasing the thickness of the epidermal layer of the skin, cosmetic composition.
5. The cosmetic composition according to claim 1, wherein the skin whitening is achieved by promoting the extracorporeal discharge of melanin.
6. The cosmetic composition according to claim 1, wherein the skin whitening is performed by inhibiting melanin secretion.
7. The cosmetic composition according to claim 1, wherein the composition promotes hair growth.
8. According to claim 1, The vitamin K3 (menadione) is a cosmetic composition comprising 0.01 to 1% based on the total weight of the cosmetic composition.
9. According to claim 1, The composition is a cosmetic composition that exhibits resistance to trypsin.
10. Vitamin K3 (menadione) or a pharmaceutical composition for skin regeneration or skin whitening comprising a pharmaceutically acceptable salt thereof as an active ingredient.
11. A quasi-drug composition for skin regeneration or skin whitening, comprising vitamin K3 (menadione) or a pharmaceutically acceptable salt thereof as an active ingredient.
12. Vitamin K3 (menadione) or a pharmacologically acceptable salt thereof as an active ingredient, for skin regeneration or skin whitening health functional food.

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