KR102275267B1 - Composition for improving skin - Google Patents

Abstract

The present invention relates to a composition for improving skin. Tetrahydropalmatine according to the present invention exhibits an antioxidant effect by scavenging free radicals, has an anti-inflammatory effect by inhibiting NO production, and exhibits a whitening effect by inhibiting melanin synthesis, as well as excellent activity of PPAR alpha Thus, it has an atopic improvement effect, and it can be used as a pharmaceutical composition, external preparation for skin, cosmetic composition or health food by showing an effect of improving skin moisture due to increased filaggrin expression and skin texture improvement by improving skin brightness. In addition, it promotes the proliferation of skin stem cells and exhibits a stem cell maintenance effect, so it can be used as a composition for promoting stem cell activity and a cosmetic comprising the same.

Description

Composition for improving skin

The present invention relates to a composition for improving skin having effects such as atopy improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement.

Active oxygen introduced from outside the living body or generated in the living body causes many problems, such as promoting aging of the living body or causing cancer. Therefore, a lot of development and research on antioxidants that inhibit oxidation by active oxygen are being made. Antioxidants are widely distributed in animals and plants, and many phenolic compounds, flavonoids, tocopherols, vitamin C, and selenium are known in fruits and vegetables. However, antioxidant substances present in nature cannot be expected to have a substantially sufficient effect when applied to the skin. Therefore, although many synthetic antioxidants have excellent antioxidant power and are inexpensive, their use is limited due to safety concerns such as side effects in the human body.

In addition, inflammation is an immune response of the human body in response to a wound or disease, and oxidative stress such as ultraviolet rays, free radicals, and free radicals activate inflammatory factors, thereby causing various diseases and aging of the skin. Vasoactive polypeptides such as kinin, plasmin, and complement act as vasodilators, contractiles, and chemotaxis, and in addition, lymphocytic such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized to prostaglandin and leukotriene, which are inflammatory mediators, through two pathways, cyclooxygenase or lipooxygenase, and mediates various inflammatory responses.

On the other hand, in order to eliminate inflammation, an anti-inflammatory agent that acts to remove an inflammatory source, reduce biological reactions and symptoms. Substances used for anti-inflammatory purposes so far include flufenamic acid, ibuprofen, benzydamine, indomethacin, etc. as a non-steroidal type, and prednisolone as a steroid type. , and dexamethasone. In addition, allantoin, azene, hydrocortisone, etc. are known to be effective in anti-inflammatory, but these substances have limitations in usage due to safety issues for the skin, or have insignificant effects, so that no inflammatory relief effect can be expected. There is this.

In addition, the epidermis, which is located at the outermost part of the skin, performs a protective function of protecting against various external physical, chemical and mechanical stimuli and preventing excessive dissipation of body moisture through the skin. This protective function is made possible by the normal formation and maintenance of the stratum corneum composed of keratinocytes. Keratinocytes are cells that are formed through gradual changes in shape and function as basal cells that have continuously proliferated in the stratum basale move to the stratum corneum, and after a certain period of time, old keratinocytes The forming cells are eliminated from the skin, and the process of epidermis differentiation or keratinization is repeated in which new keratinocytes that rise from the lowermost layer of the epidermis take over their functions. In this keratinization process, keratinocytes generate natural moisturizing factor (NMF) and intercellular lipids such as ceramide, cholesterol, and fatty acids, and the stratum corneum acts as a blocking layer from the outside, thereby creating a skin barrier. retain the function as

In addition, dry skin, which is considered one of the major diseases in modern society, is one of the symptoms caused by abnormal skin barrier function. Recently, environmental pollution, an increase in dry environments such as apartments and high-rise buildings, an increase in social stress, and unique to our country It is increasing more and more due to various factors such as excessive bathing culture, skin aging, etc., and the number of cases requiring treatment due to severe symptoms is also continuously increasing.

Recently, atopic dermatitis, which occurs in 10% of children, is also known to be the main cause of dry skin, more fundamentally, abnormal skin barrier function. In order to treat such atopic dermatitis, conventionally, many studies have been conducted to supply moisture from the outside or to minimize moisture loss from the body by focusing on maintaining adequate moisture in the skin, and in fact, ceramides ( Moisturizers such as ceramide) or derivatives thereof have been developed and are widely used in pharmaceutical or cosmetic fields.

However, most of the use of such a moisturizer is only a temporary relief of symptoms rather than a fundamental treatment, and thus does not exhibit sufficient effect on the treatment of dry skin including atopic dermatitis and skin barrier function abnormalities. Accordingly, there is an urgent need to develop a material that fundamentally regenerates the damaged skin barrier.

Recently, when PPAR alpha (Peroxisome Proliferator Activated Receptor-alpha) present in keratinocytes binds to its ligand and is activated, it is known that it has the effect of reconstructing the damaged skin barrier by promoting the differentiation of keratinocytes. In fact, when Clofibrate [Non-Patent Document 1] or WY14643 [Non-Patent Document 2], which are known agonists of PPAR alpha, is applied to skin with damaged skin barrier, differentiation of keratinocytes is promoted It has been confirmed that the recovery of the skin barrier is accelerated.

Also, to have fair and fair skin is the unchanging desire of everyone. It is genetically determined according to the concentration and distribution of melanin in the skin of a person, but it is also affected by environmental or physiological conditions such as solar ultraviolet rays, fatigue, and stress. Melanin is produced through a non-enzymatic oxidation reaction after the enzyme tyrosinase acts on tyrosine, a kind of amino acid, to convert it into DOPA and dopaquinone. As such, the pathway by which melanin is made is known, but the mechanism that induces melanin synthesis, which is the previous step of tyrosinase action, is still not fully elucidated.

On the other hand, as a general known whitening ingredient, substances that inhibit tyrosinase enzyme activity such as kojic acid, arbutin, etc., hydroquinone, vitamin-C (L-Ascorbic acid) and derivatives thereof and various There are plant extracts. By inhibiting the synthesis of melanin pigment, it is possible not only to realize skin whitening by brightening the skin tone, but also to improve skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or heredity. However, when applied to the skin, there is a problem in that the amount of use is limited due to safety issues such as irritation and redness, or the actual effect cannot be expected because the effect is insignificant.

In addition, the skin texture, which is the shape of the skin surface, is characterized by a three-dimensional microstructure formed by fine lines and is significantly different by age and region. The skin texture is divided into 1st (about 20-100 μm), 2nd (about 5-40 μm), and 3rd (about 0.5 μm) lines according to the depth of the line. It has a characteristic that polygons and star formations formed by the intersection of lines forming a clear pattern [Non-Patent Document 3].

However, it is known that with age or as the skin is damaged, the network structure of dense fine lines collapses, causing fine lines (secondary, tertiary, etc.) to disappear and the primary line to become deeper, creating wrinkles [non-patented] Literature 4]. In other words, wrinkles, a typical symptom of aging skin, are the accumulation of minute changes that have already occurred over a long period of time.

It is known that all components of human skin are embryologically derived from ectoderm or mesoderm. The epidermis, hair follicles, sebaceous and sweat glands are derived from the ectoderm, while melanocytes, nerves and special sensory receptors are derived from the neuroectoderm. Depending on the embryonic period, the stem cells of the embryo repeatedly differentiate to become cells with characteristics suitable for each tissue function, and even after becoming an adult, a certain number of stem cells remain in the tissue. In the skin, stem cells are mainly present in two places. . The first is in the hair follicle. This is a cell before cell differentiation and is known to play an important role in the regeneration of the epidermis, and plays an important role in the regeneration and growth of hair. The second is the basal layer of the epidermis. It has been found that the stem cells located here play an important role in protecting skin health by controlling not only the epidermis but also the fibroblasts of the dermis layer. The stem cells here are widely used in skin stem cell research because they have the advantage of being relatively abundant and easily obtainable. The skin is continuously regenerated, and stem cells present in the epithelium of the skin, that is, epidermal stem cells of the skin, are involved in the repair of the epithelium after a wound [Non-Patent Document 5]. The epithelial stem cells of the skin are also referred to as skin stem cells, and when the skin stem cells are activated, it can treat skin wounds such as trauma or promote wound healing, and skin wrinkle relief can also be expected. have. Integrin β1 and integrin α6 are used as indicators of skin stem cells, and it is reported that the maintenance of skin stem cells necessary for epithelial morphogenesis and differentiation is regulated by p63 protein.

Skin stem cells play an important role in maintaining skin health and physiological and biochemical homeostasis. Stem cells present in the skin also have malfunctions due to the effects of aging, which causes various problems as the homeostasis of the skin is broken. Therefore, it will be possible to improve various phenomena of skin aging through the activity of these stem cells [Non-Patent Document 6].

In addition, it is safe to the body, and the active ingredients are stable, and above all, atopy improvement, antioxidant, anti-inflammatory, moisturizing, whitening, which is superior to the existing substances that have the effect of improving atopic dermatitis, antioxidant, anti-inflammatory, moisturizing, whitening, and skin texture. However, there is an urgent need for the development of ingredients with skin texture improvement activity.

Feingold et al., J. Invest. Dermatol., 110, pp 368-375, 1998. Feingold et al., J. Invest. Dermatol., 115, pp 353-360, 2000. Journal of the European Academy of Dermatology and Venereology. 12:103-114, 1999 The British journal of dermatology. 110: 129-138, 1984, Skin research and technology. 5:189-194, 1999 Epidermal Stem Cells of the Skin, Cedric Blanpain, Elaine Fuchs, Annual Review of Cell and Developmental Biology, November 2006, Vol. 22, Pages 339-373 Human skin stem cells and the aging process, Catherin Niemann, Stem Cell Aging and Regenerative Medicine, November 2008, Pages 986-997

Accordingly, the present inventors found that tetrahydropalmatine exhibits an antioxidant effect by scavenging free radicals of fibroblasts in the skin, has an anti-inflammatory effect by inhibiting NO production, and inhibits melanin synthesis to exhibit a whitening effect as well as , PPAR alpha also has excellent activity to improve atopic dermatitis. In addition to improving skin moisture due to increased filaggrin expression, skin texture improvement effect by improving skin brightness was confirmed, stem cell proliferation promoting effect and stem cell maintenance effect By confirming the present invention was completed.

Accordingly, it is an object of the present invention to provide a composition for improving atopic dermatitis, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture comprising a compound represented by the following formula (1) as an active ingredient:

[Formula 1]

In addition, another object of the present invention is to provide a composition for promoting stem cell activity comprising a compound represented by the following formula (1) as an active ingredient, and a cosmetic composition comprising the composition:

[Formula 1]

As a means for solving the above problems, the present invention

It provides a skin external preparation for improving atopic dermatitis, antioxidant, anti-inflammatory, moisturizing, whitening and improving skin texture, comprising a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

As another means for solving the above problems, the present invention

Provided is a cosmetic composition for improving atopic dermatitis, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture comprising a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

As another means for solving the above problems, the present invention

It provides a composition for promoting stem cell activity comprising a compound represented by the following formula (1) as an active ingredient, and a cosmetic composition comprising the composition.

[Formula 1]

Tetrahydropalmatine according to the present invention exhibits an antioxidant effect by scavenging free radicals, has an anti-inflammatory effect by inhibiting NO production, and exhibits a whitening effect by inhibiting melanin synthesis, as well as excellent activity of PPAR alpha Thus, it has an atopic improvement effect, and it can be used in pharmaceuticals, cosmetics, or health food because it exhibits an effect of improving skin moisture due to increased filaggrin expression and improvement of skin texture due to improvement of skin brightness. In addition, it promotes the proliferation of skin stem cells and exhibits a stem cell maintenance effect, so it can be used as a composition for promoting stem cell activity and a cosmetic comprising the same.

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

In order for atopic improvement, antioxidant, anti-inflammatory, moisturizing, whitening, and skin texture improvement ingredients to exhibit excellent effects when applied to actual skin, high-active atopic improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement activities at low concentrations are required. It has excellent ability to penetrate and absorb the skin, and has low volatility so that it can stay for a sufficient time to exhibit the effects of improving atopy, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture, and the active ingredient is stable on the composition or on the skin. It is desirable that it is maintained in a healthy state, is easy to formulate into medicines or cosmetics, and is safe for the skin. However, it is rare among known components that satisfy all of the above properties. For example, some atopic improvement, antioxidant, anti-inflammatory, moisturizing, whitening, and skin texture improving ingredients have excellent atopy improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement activities even at low concentrations in in vitro experiments. It has poor ability to penetrate and absorb, making it difficult to apply to actual skin. Another active ingredient has low hydrophilicity, making it difficult to formulate into medicines or cosmetics. In addition, some atopy improving, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improving ingredients are decomposed or transformed into other compounds when exposed to heat, light, or oxygen, and their effects disappear before application to the skin. In some cases.

As can be seen in the following examples, tetrahydropalmatin exhibits excellent antioxidant effect, anti-inflammatory effect, melanin production inhibitory effect, PPAR alpha activation effect, filaggrin expression effect, skin brightness improvement effect at low concentration, thus improving atopic dermatitis , antioxidant, anti-inflammatory, moisturizing, skin whitening and skin texture improvement, it can be used as an active ingredient in pharmaceuticals, cosmetics, and health foods.

Therefore, the present invention provides a pharmaceutical composition for improving atopic dermatitis, antioxidant, anti-inflammatory, moisturizing, skin whitening, and skin texture improvement comprising a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

The compound name of the compound of Formula 1 above is (13aS)-2,3,9,10-tetramethoxy-6,8,13,13a-tetrahydro-5H-isoquinolino[2,1-b]isoquinoline[ It is called (13aS)-2,3,9,10-tetramethoxy-6,8,13,13a-tetrahydro-5H-isoquinolino[2,1-b]isoquinoline], also called tetrahydropalmatine.

The compound of Formula 1 may be synthesized and used, or a commercially available compound may be used.

The pharmaceutical composition for improving atopic dermatitis, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture of the present invention may include a pharmaceutically acceptable salt of the compound of Formula 1 above.

The pharmaceutically acceptable salt of the compound of Formula 1 may be an acid addition salt formed using an organic acid or an inorganic acid, and the organic acid is, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid , malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, dichloroacetic acid, aminooxyacetic acid , benzenesulfonic acid, p-toluenesulfonic acid and methanesulfonic acid salts, and inorganic acids include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid salts. Preferably, it may be in the form of hydrochloride or acetate, and more preferably in the form of hydrochloride.

The above-mentioned acid addition salt can be prepared by a) directly mixing the compound of formula 1 and an acid, b) dissolving one of them in a solvent or a hydrous solvent and mixing, or c) dissolving the compound of formula 1 in a solvent or water. It is prepared by a general method of preparing salts by placing them in an acid in a solvent and mixing them.

In addition to the above, additional salts available include gaba salt, gabapentin salt, pregabalin salt, nicotinate salt, adipate salt, hemimalonate, cysteine salt, acetylcysteine salt, methionine salt, arginine salt, lysine salt, ornithine salt, Aspartate, etc.

In addition, when the compound of Formula 1 of the present invention is used as a pharmaceutical, it may further contain one or more active ingredients exhibiting the same or similar function. For example, it may include known atopic improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement ingredients. When additional atopic improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement components are included, the atopic improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement effects of the composition of the present invention may be further enhanced. When adding the above ingredients, skin safety according to complex use, ease of formulation, and stability of active ingredients can be considered. In one embodiment of the present invention, the composition as an antioxidant component known in the art, tocopherol, selenium, vitamin C and phenolic compounds, as a whitening component known in the art, kojic acid (Kojic acid), arbutin (Arbutin) ), etc., a substance that inhibits tyrosinase enzyme activity, hydroquinone, vitamin-C (L-Ascorbic acid) and derivatives thereof, and various plant extracts. One or two or more components selected from the group are added. can be included as Additional ingredients may be included in an amount of 0.0001 wt% to 10 wt% based on the total weight of the composition, and the content range may be adjusted according to requirements such as skin safety and ease of formulation of the compound of Formula 1 .

In addition, the pharmaceutical composition for improving atopy, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture of the present invention may further include a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain various ingredients such as buffers, sterile water for injection, plain saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates, and the like.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and may be administered in the form of an over-the-counter pharmaceutical formulation, for example, oral and parenteral various formulations during clinical administration. , extenders, binders, wetting agents, disintegrants, and diluents or excipients such as surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the pharmaceutical composition of the present invention, for example, starch, calcium carbonate, It may be prepared by mixing sucrose or lactose, gelatin, and the like.

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

In the present invention, the term 'atopic improvement effect' refers to preventing and treating skin diseases such as atopic dermatitis and dry skin, and inhibiting, inhibiting, or alleviating atopic symptoms.

In the present invention, the term 'antioxidative effect' refers to the effect of intracellular metabolism or oxidative stress caused by UV rays, which are highly reactive free radicals or reactive oxygen species (ROS). It refers to inhibiting oxidation, and includes reducing damage to cells by removing free radicals or reactive oxygen species.

In the present invention, the term 'anti-inflammatory effect' refers to suppressing inflammation, and the inflammation is one of the defense responses of a living tissue to a certain stimulus, which causes tissue deterioration, circulatory disorder and exudation, and tissue proliferation. complex lesions. More specifically, inflammation is part of innate immunity, and as in other animals, human innate immunity recognizes patterns on the surface of cells that are specific to pathogens. Phagocytes recognize cells with such a surface as non-self and attack pathogens. If pathogens break through the body's physical barriers, an inflammatory response occurs. The inflammatory response is a non-specific defense action that creates a hostile environment for microorganisms that invade the wound site. In the inflammatory response, when there is a wound or an external infectious agent enters the body, the leukocytes responsible for the initial stage immune response flock and express cytokines. Therefore, the expression level of intracellular cytokines is an indicator of inflammatory response activation. Examples of skin diseases related to inflammation include atopic dermatitis, psoriasis, erythematous diseases triggered by radiation, chemicals, burns, etc., acid burns, bullous dermatosis, lichenoid type disease, itching caused by allergies, seborrheic eczema, acne on roses, pemphigus vulgaris, erythema multiforme, erythema nodosum, balanitis, vulvitis, inflammatory hair loss such as alopecia areata, cutaneous T-cell lymphoma, and the like.

In the present invention, the 'moisturizing effect' refers to inhibiting or suppressing a decrease in moisture in the skin or increasing the moisture content of the skin to smooth the skin surface and to impart gloss.

In the present invention, the 'whitening effect' refers to not only brightening skin tone by inhibiting the synthesis of melanin pigment, but also improving skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones or heredity.

In the present invention, the 'skin texture improvement effect' refers to smoothing the skin surface, imparting gloss, and brightening the skin tone by suppressing or inhibiting the roughening of the skin surface due to the effects of aging, stress, etc. .

When the pharmaceutical composition of the present invention contains an effective amount of the compound of Formula 1, it may provide desirable atopic improvement effects, antioxidant effects, anti-inflammatory effects, moisturizing effects, whitening effects, and skin texture improvement effects. In the present invention, the term 'effective amount' refers to a compound capable of improving skin dryness such as atopic dermatitis, inhibiting or alleviating cell oxidation, inhibiting inflammation, improving skin texture, moisturizing effect, and whitening effect means the amount of The effective amount of the compound of Formula 1 included in the composition of the present invention will vary depending on the form in which the composition is commercialized, the method in which the compound is applied to the skin, and the time it stays on the skin. For example, when the composition is commercialized as a pharmaceutical, the compound of Formula 1 may be included at a higher concentration than when commercialized as a cosmetic that is routinely applied to the skin. Accordingly, the daily dose is 0.1 to 100 mg/kg, preferably 30 to 80 mg/kg, and more preferably 50 to 60 mg/kg, based on the amount of the compound of Formula 1, and 1 to It can be administered 6 times.

The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

The present invention may also be provided in the form of an external preparation for atopic dermatitis, antioxidant, anti-inflammatory, moisturizing, whitening, and skin texture improvement including the compound of Formula 1 as an active ingredient.

When the compound of Formula 1 is used as an external preparation for skin, in addition, a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant , water, ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or common for external applications for the skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients used as a dermatome. In addition, the ingredients may be introduced in an amount generally used in the field of dermatology.

When the compound of Formula 1 is provided as an external preparation for skin, the present invention is not limited thereto, but may have a formulation such as an ointment, a patch, a gel, a cream, or a spray.

The present invention may also be provided as a formulation for atopic improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement cosmetic comprising the compound of Formula 1 as an active ingredient.

When the compound of Formula 1 is used as a cosmetic, cosmetics prepared by containing the compound of Formula 1 as an active ingredient may be prepared in the form of general emulsified formulations and solubilized formulations. For example, lotions such as softening lotions or nourishing lotions, emulsions such as facial lotions and body lotions, creams such as nourishing creams, moisture creams and eye creams, essences, cosmetic ointments, sprays, gels, packs, sunscreens, makeup bases, liquid It may have formulations such as foundation, powder, cleansing cream, cleansing lotion, and makeup remover such as cleansing oil, cleansing foam, soap, body wash, and the like.

In addition, the cosmetic is a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water in addition to the compound of Formula 1 , ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or commonly used in cosmetics It may contain adjuvants commonly used in the field of cosmetology, such as any other ingredients.

In the case of wash-off type cosmetics such as makeup removers and detergents, in which the active ingredient stays on the skin within a short period of time when the compound of Formula 1 is commercialized as a cosmetic, a relatively high concentration of the compound of Formula 1 is used. could include On the other hand, in the case of leave-on type cosmetics, such as lotions, emulsions, creams, and essences, in which the active ingredient stays on the skin for a long period of time, a lower concentration of the compound of Formula 1 compared to wash-off type cosmetics would be free to include. Although not limited thereto, in one embodiment of the present invention, the composition may contain the compound of Formula 1 in an amount of 0.0001% to 10% by weight (preferably 0.0001% to 1% by weight) based on the total weight of the composition. can When the composition of the present invention contains the compound of Formula 1 in an amount of less than 0.0001% by weight, sufficient atopic improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement effects cannot be expected, and it may contain more than 10% by weight. In some cases, this is to prevent unwanted reactions such as allergies or problems with skin safety.

The present invention also relates to a health food for improving atopic dermatitis, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture, comprising the compound of Formula 1 above.

As used herein, the term 'health food' refers to food prepared by adding the compound of Formula 1 to food materials such as beverages, teas, spices, gum, and confectionery, or encapsulating, powdering, suspension, etc. It means to bring a specific effect, but unlike general drugs, it has the advantage that there are no side effects that may occur when taking the drug for a long period of time by using food as a raw material.

Since the health food of the present invention obtained in this way can be consumed on a daily basis, it is very useful because it can be expected to have high atopy improvement, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture improvement effects.

When the compound of Formula 1 is used as a food additive, the compound of Formula 1 may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). In general, in the production of food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. .

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all health foods in the ordinary sense.

The health beverage composition of the present invention may contain various flavoring agents or natural carbohydrates as an additional component like a conventional beverage. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumatine and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.

In addition to the above, the health food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol , a carbonation agent used in carbonated beverages, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also provides a composition for promoting stem cell activity comprising the compound of Formula 1 as an active ingredient.

In the present invention, the term 'stem cell' refers to a cell capable of self-dividing and having the ability to differentiate into a wide variety of specific cell types. The type of these stem cells is not particularly limited, and in one embodiment, the stem cells may be skin stem cells. The 'skin stem cells' refer to stem cells that can be differentiated into cells constituting the skin (epidermis, dermis, and subcutaneous fat layer). Cells constituting the skin include keratinocytes, melanocytes, and fibroblasts (mainly responsible for the biosynthesis of collagen and elastin) present in the epidermis and dermis.

The type of the skin stem cells is not particularly limited. The skin stem cells used in the present invention can be used regardless of where they originate. For example, the skin stem cells may be obtained from a known source of skin stem cells, for example, hair follicles or the basal layer of the epidermis, and the animal to be harvested may be a mammal. In one embodiment, a mammal can include, but is not limited to, a human, mouse, rat, guinea pig, rabbit, monkey, pig, horse, cow, sheep, antelope, dog or cat. Preferably the mammal may be a human. Such, a method for obtaining skin stem cells from a skin stem cell source is well known in the art.

In the present invention, the 'stem cell activity promoting effect' refers to including a stem cell proliferation promoting effect and/or stem cell maintenance effect. Skin stem cells play an important role in maintaining skin health and physiological and biochemical homeostasis. Stem cells present in the skin also have malfunctions due to the effects of aging, which causes various problems as the homeostasis of the skin is broken. Various phenomena of skin aging can be improved through the activity of these stem cells [Non-Patent Document 6], and the present invention confirmed the effects of improving atopy, antioxidant, anti-inflammatory, moisturizing, whitening and skin texture.

Therefore, the present invention can provide a cosmetic composition comprising the composition for promoting stem cell activity. This cosmetic formulation is the same as described in the description of the cosmetic formulation containing the compound of Formula 1 above.

Hereinafter, the present invention will be described in detail by way of Examples. However, the following examples only illustrate the present invention, and the content of the present invention is not limited to the following examples.

Reference example 1: tetrahydro palmatin ( Tetrahydropalmatine ) of the substance information

[Formula 1]

CAS No.: 10097-84-4

Where to buy: Sichuan Hongjie Import&Export CO., LTD

Reference example 2: serum free in the medium condition Cultivation of skin stem cells

Human epidermal stem cells purchased from Cellntec were placed in a 48-well plate (6×10 ³ cells/well), and BPE (Bovine pituitary extract) similar to fetal bovine serum was added. CNT-57 medium (Cellntec) was used for 24 hours _{and cultured at 5% CO 2} and 37° C. conditions. Thereafter, the culture medium was removed through a suction tube, and then the medium components were removed using a PBS solution (GibcoBRL), and 20 μg/mL of the compound of Formula 2 was treated with BPE-free CNT-57 medium to 5% CO Incubated for 72 hours at _{2 and 37 ℃ conditions.}

Experimental example One: CCK Effect of promoting skin stem cell proliferation through the -8 evaluation method

CCK-8 (Cell counting kit-8) was evaluated on the skin stem cells cultured in Reference Example 2. CCK-8 evaluation is an analysis method that indirectly indicates the density of living cells by measuring the absorbance of Formazan, which is generated by dehydrogenase in the intracellular electron transport system by decomposing Tetrazolium Salt.

Cells were treated with a CCK-8 solution (treated with 1/10 of the medium volume) at 37° C. for 2 hours, and then spectroscopically measured by measuring absorbance at a wavelength of 450 nm. The proliferation-promoting effect of the compound of Formula 1 was obtained by calculating the proliferation rate (%) according to the following Equation 1 compared to the control (CNT-57 medium with BPE added) from the measured absorbance value, and the values are shown in Table 1 below. .

[Equation 1]

Growth rate (%) = (absorbance of sample treatment group / absorbance of control group) × 100

Skin stem cell proliferation promoting effect additive sample density Cell proliferation rate (%) compared to control compound of formula 1 20 μg/mL 125% control - 100%

As can be seen from the results in Table 1, the skin stem cells in the medium condition treated with the compound of Formula 1 of the present invention exhibited an excellent proliferation-promoting effect.

Experimental example 2: Stem cell nature of skin stem cells ( stemness ) check the retention effect

In order to confirm the stem cell maintenance effect of the skin stem cells cultured in Reference Example 2, the expression level of p63 was evaluated. p63 is an intracellular transcription factor and is well known as a maintenance marker of skin stem cells. Cell RNA was isolated to synthesize cDNA, and then real-time-PCR was performed with Taqman staining solution to measure the expression level of p63. In this experiment, the concentration of RNA was normalized to S16 ribosomal RNA.

The experimental results are shown in Table 2 below. In Table 2 below, the value of the p63 increase fold means the fold compared to the control (CNT-57 medium with BPE added).

Stemness maintenance effect (number of repetitions = 3) additive sample density p63 multiplier (fold) compound of formula 1 20 μg/mL 1.228 control - 1.0

As can be seen from the results in Table 2, the medium condition treated with the compound of Formula 1 of the present invention promotes p63 expression in skin stem cells, and thus has excellent stem cell maintenance effect.

Example 1: Whitening effect - Confirmation of melanin production inhibitory effect

The whitening effect at the cellular level was tested by adding the compound of Formula 1 to the culture medium of B-16 mouse melanoma cells. (Lotan R., Lotan D. Cancer Res. 40:3345-3350, 1980).

Before the experiment, toxicity was evaluated for melanoma cells in mice, and a non-toxic concentration was selected for whitening evaluation.

The compound of Formula 1 was tested to have final concentrations of 10㎍/mL and 20㎍/mL in the culture medium, and arbutin, a control, was added to the medium so that it became 200㎍/mL and treated with B-16 melanoma cells, respectively. Incubated for one day.

Thereafter, the cells were treated with trypsin, removed from the culture vessel, centrifuged, and then melanin was extracted. To the detached cells, 1 mL of sodium hydroxide solution (1N) was added and boiled for 10 minutes to dissolve melanin. Using a spectrophotometer, absorbance was measured at 400 nm to measure the amount of melanin produced.

The amount of melanin was measured by a method indicated by the absorbance per unit cell number (10 ⁶ cells), the relative melanin production to the control was calculated as the inhibition rate (%), and the results are summarized in Table 3 below.

Melanogenesis inhibitory effect at the cellular level according to the concentration (number of repetitions = 3) sample Inhibition rate (%) Control (no additive) - Control 1: Arbutin (200 μg/mL) 30.8 Compound of Formula 1 (20㎍/mL) 38.2 Compound of Formula 1 (10 μg/mL) 23.5

As can be seen from the results of Table 3, it can be seen that the compound of Formula 1 has a superior ability to inhibit melanin production in cultured mice melanoma cells when compared with arbutin, which is a known whitening material.

Example 2: Antioxidant effect- free radical erasure rate

In order to confirm the antioxidant activity of the compound of Formula 1, free radical scavenging activity was measured. Free radical scavenging activity was measured using DPPH. DPPH was purchased from Sigma Co., Ltd. (USA) and used. First, a standard DPPH ethanol solution of 1.5 mM (0.06 mg/mL) was prepared. Then, ethanol was added to each of the compound of Formula 1 and ascorbic acid, an antioxidant as a reference material, to prepare samples at concentrations of 50 μg/mL, 25 μg/mL, 12.5 μg/mL, 6.25 μg/mL, and 3.125 μg/mL. . Then, the sample and the standard DPPH solution were added in the same ratio, stirred well, reacted at 37° C. for 30 minutes, and absorbance was measured at 520 nm. At this time, the addition of ethanol instead of the sample was used as a control. _{The free radical scavenging ability was obtained by calculating the IC 50} , which is the half maximal inhibitory concentration (median inhibition value), and the results are shown in Table 4 below. IC ₅₀ is the concentration of ascorbic acid and the compound of Formula 1 required to remove 50% of free radicals of the no-addition control group, and is a general method of expressing the free radical scavenging ability.

Free radical scavenging rate (IC ₅₀ ) sample Free radical scavenging rate (㎍/mL) ascorbic acid 5.2 compound of formula 1 3.4

As can be seen from the results of Table 4, it can be seen that the compound of Formula 1 exhibits high activity compared to ascorbic acid, which is a known antioxidant, and thus has an antioxidant effect.

Example 3: Anti-inflammatory effect- NO production inhibitory effect

In order to confirm the anti-inflammatory effect and skin trouble alleviation effect of the compound of Formula 1, a nitric oxide (NO) formation inhibition experiment was performed by the GRIESS method using the RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, which are macrophages of mice, were subcultured several times, ^{and placed in a 24-well plate so that 3×10 5} cells were put into each well, and then cultured for 24 hours. Then, it was replaced with a cell medium containing the compound of Formula 1 diluted to the concentration shown in Table 5 below. At this time, 20 μg/mL of L-NMMA (L-NG-Monomethylarginine), which is a NO-production inhibitor, was treated together as a positive control and incubated for 30 minutes, and LPS (Lipopolysaccharide) was treated as a stimulant by 1 μg for 24 hours. incubated during 100 μl of the supernatant was taken and transferred to a 96-well plate, 100 μl of GRIESS solution was added and reacted at room temperature for 10 minutes, and the NO inhibitory effect of Compound 1 was determined by measuring the absorbance at 540 nm, and the results are shown below. Table 5 shows.

NO production inhibitory effect sample NO production inhibition rate (%) L-NMMA (positive control) 41.53 Compound of Formula 1 (0.1%) 34.76 Compound of formula 1 (1%) 41.25

As can be seen from the results of Table 5, it can be seen that the compound of Formula 1 has an ability to inhibit NO production at the same level as that of L-NMMA, which is a known anti-inflammatory material.

Example 4: PPAR Verification of alpha activation promotion effect

In order to confirm the PPAR alpha activation promoting effect of the compound of Formula 1, the following experiment was performed using the compound of Formula 1. C2C12 cells (ATCC CRL-1772), a mouse-derived myoblast cell line, were subcultured in DMEM (Dulbeco's Modified Eagle's Medium) medium containing 10% fetal bovine serum. _{The vector used here is 1) encoding Ser 167} to Tyr ₄₆₈ in the yeast transcription factor GAL4-DBD (DNA Binding Domain) and human PPAR alpha LBD (Ligand Binding Domain) in the SV40 promoter of the pZEO vector (Invitrogen). A vector containing a DNA fragment (gene bank information, NM 005036), 2) a vector in which the GAL4 response sequence is inserted into the multiple restriction enzyme recognition site of a pGL3 vector (Promega) expressing luciferase (Luciferase), and 3) trans A vector expressing β-galactosidase was used as a transfection internal control.

The cultured cells ^{were plated on a 24-well plate at a concentration of 4×10 4} , cultured for 12 hours, and the three types of plasmid genes were transiently transfected using Lipopectamine. . After 8 hours, the compound of Formula 1 was treated and incubated for 12 hours, washed with 1×PBS and lysed with 1×reporter lysis buffer, followed by a luciferase assay kit (Promega) and The activity of luciferase was measured using a β-galactosidase assay kit (Promega). That is, PPAR alpha activity was measured as 'luciferase activity/β-galactose activity'.

In this experiment, as a positive control, Wy-14,643 (Calbiochem), which is known to be the strongest of the ligands of PPAR alpha, was used, and as a negative control, 0.05% of DMSO used for dissolving the sample was used.

According to the above experimental method, C2C12 cells were treated with the compound of Formula 1 to measure the activity of PPAR alpha according to the concentration change, and the results are shown in Table 6 below. In Table 6 below, the value of PPAR alpha activity means a percentage compared to the negative control (DMSO 0.05%).

PPAR alpha activity sample PPAR alpha activity (%) DMSO (0.05%) 100 Wy-14,643 (0.5 μM) 280 Wy-14,643 (1 μM) 935 compound of formula 1 (0.05%) 694 compound of formula 1 (0.5%) 940

As shown in the results of Table 6, it was confirmed that the compound of Formula 1 exhibits very strong PPAR alpha activity when compared with the positive control, which is a purified compound.

Example 5: of Filaggrin Expression promoting effect

The compound of Formula 1 was added to the culture medium of the human keratinocyte strain HaCaT cells at a concentration of 5 μg/mL and 10 μg/mL, respectively, and cultured for 1 day, then RNA of the cells was isolated and cDNA was synthesized, followed by Taqman staining solution Real-time PCR was performed with RNA concentrations were normalized to S16 ribosomal RNA.

Filaggrin expression effect according to concentration (number of repetitions = 3) additive sample Factor of increase in filaggrin compared to control group Control (no additive) 1.00 Compound of Formula 1 (5㎍/mL) 25.0 Compound of Formula 1 (10㎍/mL) 49.6

As can be seen from the results of Table 7, the compound of Formula 1 has the ability to promote filaggrin expression in the human keratinocyte strain HaCaT cells.

Example 6: To improve keratin exfoliation (turn-over) About efficacy

The nutritional cream of Formulation Example 2 below was tested for the effect of improving keratin exfoliation on healthy women in their 20s to 50s.

After depositing a 1.5% solution of DHA (Dihydroxyacetone, Sigma Aldrich, USA) with a closed patch for 8 hours on the test site of 20 women aged 20 to 50 years, the preparation nutrient cream was applied to the test site twice daily. After 4 days of application, 8 days after application, and 12 days after application, using a Chromameter CR-400 (Minolta, Japan) to measure the brightness of the skin at the deposition site and take pictures using a DSLR. The measured values were evaluated by obtaining the average value of three times excluding the maximum and minimum values, and the higher the improvement in skin brightness at the deposition site, the greater the effect of improving keratin exfoliation.

The results are shown in Table 8 below.

Skin brightening effect division 4 days after application 8 days after application 12 days after application Improvement (%) 1.28 4.83 12.37

As can be seen in Table 8, when the formulation example nutritional cream according to the present invention was used, it was found that there was an effect of improving keratin exfoliation.

Formulation example 1: Preparation of pharmaceutical formulations

1. Preparation of tablets

0.2 mg of compound of formula 1

Corn Starch 100mg

Lactose 100mg

stearic acid Magnesium 2mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional method for manufacturing tablets.

Formulation example 2: Manufacture of cosmetics

1. Manufacture of nourishing cream

As shown in the following composition, a nutritional cream containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.2 wt% of the compound of Formula 1

Beta-1,3-glucan 5.0 wt%

Beeswax 10.0 wt%

Polysorbate 60 1.5 wt%

Sebum 60 hydrogenated castor oil 2.0 wt%

Sorbitan sesquioleate 0.5 wt%

Liquid paraffin 10.0 wt%

5.0 wt% squalane

5.0 wt% caprylic/capric triglyceride

glycerin 5.0 wt%

Butylene glycol 3.0 wt%

Propylene glycol 3.0 wt%

Triethanolamine 0.2 wt%

Preservative 0.05 wt%

dye 0.05% by weight

Flavor 0.05% by weight

Purified water to 100% by weight

Formulation example 3: external skin preparations Produce

1. Preparation of ointment

As shown in the following composition, an ointment containing the compound of Formula 1 as an active ingredient was prepared according to a conventional method.

0.5 wt% of the compound of Formula 1

Beta-1,3-glucan 10.0 wt%

Beeswax 10.0 wt%

Polysorbate 60 5.0 wt%

Sebum 60 hydrogenated castor oil 2.0 wt%

0.5 wt% of sorbitan sesquioleate

Vaseline 5.0% by weight

Liquid paraffin 10.0 wt%

5.0 wt% squalane

Shea Butter 3.0 wt%

5.0 wt% caprylic/capric triglycerides

10.0 wt% glycerin

Propylene glycol 10.2 wt%

0.2% by weight of triethanolamine

Preservative 0.05 wt%

dye 0.05% by weight

Flavor 0.05% by weight

Purified water to 100% by weight

Claims (15)

delete delete delete delete delete delete delete A cosmetic composition for moisturizing the skin comprising a compound represented by the following formula (1) as an active ingredient:
[Formula 1]

delete delete delete A cosmetic composition for improving skin texture comprising a compound represented by the following formula (1) as an active ingredient:
[Formula 1]

A cosmetic composition for skin regeneration according to the promotion of stem cell activity comprising a compound represented by the following formula (1) as an active ingredient:
[Formula 1]

14. The method of claim 13,
The stem cell activity is a cosmetic composition for skin regeneration according to stem cell activity promotion, characterized in that stem cell proliferation promotion or stem cell maintenance. delete