KR20170035661A - Composition for improving skin comprising n-Methylcytisine as active ingredient - Google Patents
Composition for improving skin comprising n-Methylcytisine as active ingredient Download PDFInfo
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- KR20170035661A KR20170035661A KR1020150134843A KR20150134843A KR20170035661A KR 20170035661 A KR20170035661 A KR 20170035661A KR 1020150134843 A KR1020150134843 A KR 1020150134843A KR 20150134843 A KR20150134843 A KR 20150134843A KR 20170035661 A KR20170035661 A KR 20170035661A
- Authority
- KR
- South Korea
- Prior art keywords
- skin
- composition
- acid
- promoting
- methylcytosine
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- CULUKMPMGVXCEI-VHSXEESVSA-N n-methylcytisine Chemical compound C12=CC=CC(=O)N2C[C@@H]2CN(C)C[C@H]1C2 CULUKMPMGVXCEI-VHSXEESVSA-N 0.000 title claims abstract description 10
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4906—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom
- A61K8/4926—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having six membered rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
Abstract
The present invention relates to a composition for improving skin. N-Methylcytisine according to the present invention is effective in promoting skin stem cell activity, promoting collagen synthesis of skin fibroblasts, promoting melanin synthesis, and improving skin brightness, thereby improving skin regeneration, Skin whitening, skin texture improvement and skin barrier strengthening, cosmetics, food or skin external formulations. In addition, it promotes proliferation of dermal stem cells and exhibits a stem cell-retaining effect, and thus can be used for the production of cosmetic formulations for promoting stem cell activity.
Description
The present invention relates to a skin improving composition exhibiting skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement, skin barrier enhancement, and stem cell activity promoting effect.
Collagen is a major substrate protein produced in fibroblasts of the skin and exists in extracellular epilepsy. Its important functions are mechanical rigidity of skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, division of cells and differentiation Growth or wound healing) are known. Such collagen is reduced by aging and photo aging caused by ultraviolet irradiation, which is known to be closely related to the wrinkling of the skin. Also, in recent years, extensive research on skin aging has developed, and important functions of collagen in skin have been revealed.
Effective ingredients promoting collagen synthesis and exhibiting wrinkle-reducing effects are known. For example, retinoic acid, transforming growth factor (TGF) [non-patent document 1], animal placenta-derived protein [Patent document 1], betulinic acid [Patent document 2], chlorella extract [ Patent Documents 3 and 4] are known as collagen synthesis promoting substances. However, the above-mentioned effective ingredients are limited in the use amount due to safety problems such as irritation and redness when applied to the skin, or have insufficient effect, so that the effect of improving the skin function by promoting the collagen synthesis of the skin can not be expected.
In addition, the epidermis located at the outermost part of the skin protects against various external physical, chemical and mechanical stimuli and protects against excessive divergence of body water through the skin. This protective function is possible by normally forming and maintaining the stratum corneum composed of keratinocytes. The keratinocyte is a cell formed by a stepwise change in morphology and function while a basal cell that continuously proliferates in the stratum basale moves to the stratum corneum, Forming cells are removed from the skin and the new keratinocytes from the epidermis's bottom layer repeat the process of epidermis differentiation or keratinization replacing its function. In this keratinization process, keratinocytes produce intercellular lipids such as natural moisturizing factors (NMF) and ceramides, cholesterol, and fatty acids, which act as barrier layers to the outside, As shown in Fig.
In addition, it is the desire of everyone to have a whiter skin. It is genetically determined by the concentration and distribution of melanin in human skin, but is also influenced by environmental or physiological conditions such as sunlight, fatigue, and stress. Melanin is produced by a nonenzymatic oxidation reaction after tyrosine, an amino acid, is converted to DOPA or dopaquinone by an enzyme called tyrosinase. Thus, the pathway through which melanin is made is known, but the mechanism by which melanin synthesis, the previous step in which tyrosinase acts, is not yet elucidated.
On the other hand, commonly known whitening ingredients include substances inhibiting the activity of tyrosinase enzymes such as kojic acid and arbutin, hydroquinone, vitamin C (L-Ascorbic acid) There are plant extracts. By inhibiting the synthesis of melanin pigment, they can brighten the skin tone to realize skin whitening, and it is possible to improve skin hypercholesterolemia such as stain or freckles due to ultraviolet rays, hormones or heredity. However, when applied to skin, there is a problem that the use amount is limited due to safety problems such as irritation and redness, or the effect is insignificant, so that a practical effect can not be expected.
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 according to age and site. The skin texture is divided into primary (about 20-100 μm), secondary (about 5-40 μm), and tertiary (about 0.5 μm) lines depending on the depth of the line. And has a characteristic that the polygons and star formation that occur when the lines meet meet [non-patent document 1].
However, as the age increases or the skin becomes damaged, the network structure of the dense fine lines collapses and fine lines (secondary, tertiary, etc.) disappear and the primary lines become deeper and wrinkles are known to be produced Literature 2]. In other words, wrinkles, a typical sign of skin aging, are accumulations of minute changes that have already occurred over a long period of time.
Embryologically, all components of human skin are known to originate from ectoderm or mesodermal lobe. Epidermis, hair follicles, sebaceous glands and glands are originated from ectoderm, and melanocytes, nerves and special sensory receptors are derived from neuroepithelial cells. According to the developmental stage, the embryonic stem cells are repeatedly differentiated and become cells with the characteristics of each tissue function. After the embryo, a certain number of stem cells remain in the tissue. . The first is in the hair follicle. It is known to play an important role in the regeneration of the epidermis into cells before cell differentiation occurs, and plays an important role in hair regeneration and growth. The second is the basal layer of the epidermis. The stem cells found here play an important role in maintaining skin health by administering not only the epidermis but also the fibroblasts of the dermal layer. The stem cells here are relatively large in quantity and easy to obtain, making them widely used in the study of skin stem cells. The skin is constantly renewed, and stem cells present in the epithelium of the skin, that is, epidermal stem cells of the skin, are involved in the recovery of the epithelium after injury [Non-Patent Document 3]. The epithelial stem cells of the skin are also referred to as skin stem cells. When the skin stem cells are activated, it is possible to treat skin wounds such as trauma, promote wound healing, have. Integrin β1 and integrin α6 have been used as indices of dermal stem cells and maintenance of dermal stemness necessary for epithelial morphogenesis and differentiation has been reported to be regulated by p63 protein .
Dermal stem cells play an important role in maintaining the health and physiological and biochemical homeostasis of the skin. Stem cells present in the skin are also abnormally functioned due to the effect of aging, and thus various problems arise as the homeostasis of the skin is broken. Therefore, it is possible to improve various phenomena of skin aging through the activation of stem cells [Non-Patent Document 4].
In addition, the present invention provides a cosmetic composition which is safe and biologically safe, has an excellent effect on skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement and skin barrier enhancement There is a desperate need for the development of a composition having an activity of enhancing, skin whitening, improving skin texture and enhancing skin barrier.
Accordingly, the present inventors have found that N-methylcytisine promotes proliferation of dermal stem cells, maintains stem cell characteristics, exhibits skin regeneration promoting effect, promotes collagen synthesis of fibroblasts of skin, Or exhibiting an effect of enhancing elasticity, exhibiting a skin barrier strengthening effect by promoting skin stem cell activity and promoting collagen synthesis, exhibiting a whitening effect by inhibiting melanin synthesis, improving skin texture by improving skin brightness, Promoting proliferation of stem cells and promoting stem cell activity promoting stem cell retention, thereby completing the present invention.
Accordingly, an object of the present invention is to provide a composition for enhancing skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement and skin barrier enhancement comprising n-methylcycin, which is a compound represented by the following formula :
[Chemical Formula 1]
Another object of the present invention is to provide a composition for promoting stem cell activity comprising n-methylcyclin, which is a compound represented by the following formula 1, as an active ingredient:
[Chemical Formula 1]
As a means for solving the above-mentioned problems, the present invention relates to a skin comprising, as an active ingredient, n-methylcytisine, which is a compound represented by the following formula (1), for the preparation of pharmaceutical, cosmetic, There is provided a composition for regenerating, regenerating, improving wrinkles, improving elasticity, whitening skin, improving skin texture and strengthening skin barrier:
[Chemical Formula 1]
As another means for solving the above problems, the present invention provides a composition for promoting stem cell activity comprising n-methylcytosine, which is a compound represented by the following formula (1), as an active ingredient for the production of cosmetic formulations:
[Chemical Formula 1]
N-Methylcytisine according to the present invention promotes proliferation of dermal stem cells, maintains stem cell characteristics, exhibits skin regeneration promoting effect, promotes collagen synthesis of fibroblasts of skin, Exhibits the effect of enhancing elasticity, exhibits a skin barrier strengthening effect by promoting skin stem cell activity and promoting collagen synthesis, exhibits a whitening effect by inhibiting melanin synthesis, and exhibits an effect of improving skin texture due to improvement in skin brightness. , Food or skin external preparation formulations.
In addition, n-methylcytosine promotes the proliferation of dermal stem cells and promotes stem cell activity that maintains stem cell characteristics, and thus can be used for the production of cosmetic formulations for promoting stem cell activity.
Hereinafter, the configuration of the present invention will be described in detail.
Skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement and skin barrier enhancement In order to exert excellent effects when the ingredient is applied to the actual skin, it is necessary to improve the skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, So as to be able to stay for a sufficient time to exhibit an effect of enhancing skin texture and enhancing skin barrier and having excellent ability to permeate and absorb the skin and to exhibit skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement, Is low in volatility, maintains active ingredients stably on the composition or on the skin, is easy to be formulated into pharmaceuticals, cosmetics, foods, skin external agents and the like, and is safe for skin. However, the components satisfying all of the above-mentioned characteristics among the known components are not common. For example, some skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement, and skin barrier enhancement ingredients can be used for skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, It has excellent reinforcing activity, but its ability to permeate through the skin is poor, making it difficult to apply to real skin. In addition, some skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin tone improvement and skin barrier enhancement ingredients may be decomposed or converted into other compounds when exposed to heat, light or oxygen, Sometimes the effect disappears.
As can be seen in the following examples, N-Methylcytisine shows remarkably superior skin stem cell activity promoting effect, collagen synthesis promoting effect, melanin formation inhibiting effect and skin brightness improvement effect at a low concentration, It can be used as an effective ingredient for pharmacy, cosmetics, food, skin external agent for regenerating, improving wrinkles, improving elasticity, whitening skin, improving skin texture and strengthening skin barrier.
Accordingly, the present invention provides a composition for enhancing skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement and skin barrier enhancement comprising n-methylcytosine, which is a compound represented by Chemical Formula 1, as an active ingredient.
[Chemical Formula 1]
The compound of Chemical Formula 1 is named (1R, 5S) -1,2,3,4,5,6-Hexahydro-3-methyl-1,5-methano-8H-pyrido [ , 5] diazocin-8-one, also called N-methylcytisine.
The above-mentioned n-methylcytosine may be synthesized, or a commercially available compound may be used.
In the present invention, the term " skin regeneration effect " refers to restoration of skin tissue against damage caused by external or internal causes of the skin. Damage due to external causes may include ultraviolet rays, external contaminants, wound, trauma, etc. The damage caused by the internal causes may be stress. Preferably, the skin regeneration is associated with dermal stem cells which play an important role in maintaining the health and physiological and biochemical homeostasis of the skin. The maintenance of the skin stem cell state necessary for epithelial morphogenesis and differentiation is regulated by the p63 protein do. Accordingly, by treating the active ingredient of the present invention, the skin regeneration effect can be remarkably enhanced by promoting the proliferation of the dermal stem cells and maintaining the dermal cellularity.
In the present invention, the term " wrinkle-reducing effect " refers to inhibiting or inhibiting the generation of wrinkles on the skin, or alleviating already-generated wrinkles.
In the present invention, the 'elasticity-enhancing effect' refers to an increase in elasticity to the skin, which suppresses or inhibits the loss of elasticity of the skin, or alleviates the already-reduced elasticity.
Preferably, the wrinkle-improving and elasticity-enhancing effects are related to the mechanical rigidity of the skin, the resistance of the connective tissue and the binding force of the tissue, the support of cell adhesion, and the collagen inducing cell division and differentiation. Therefore, by treating the active ingredient of the present invention, collagen synthesis can be promoted and the effect of improving wrinkles and improving the elasticity can be remarkably improved.
In the present invention, the 'whitening effect' refers to not only brightening the skin tone by inhibiting the synthesis of the melanin pigment but also improving skin hypercholesterolemia due to ultraviolet rays, hormones or heredity, such as spots or freckles.
In the present invention, the term " skin texture improving effect " refers to smoothing the skin surface, imparting gloss, and brightening the skin tone by inhibiting or inhibiting roughness of the skin surface due to aging, stress, .
In the present invention, the 'skin barrier' is a skin barrier that performs a defense function against various physical, chemical and mechanical stimuli of the outside. The horny layer formed by repeated epidermal differentiation or keratinization acts as a barrier layer against the outside, It means to function. Thus, in the present invention, the term 'skin barrier strengthening effect' refers to a function of promoting collagen synthesis of fibroblasts of the skin and promoting proliferation of skin stem cells, thereby promoting skin regeneration, improving wrinkles, To make healthy.
For regenerating skin, improving wrinkles, improving elasticity, skin whitening, improving skin texture and strengthening skin barrier of the present invention The compositions may be used for the manufacture of cosmetic formulations.
The cosmetic formulation may be in the form of a conventional emulsion formulation and a solubilized formulation. For example, creams, essences, cosmetic creams, sprays, gels, packs, sunscreens, make-up bases, liquids such as lotions such as lotion, facial lotion, body lotion, A powder, a cleansing lotion, a makeup removing agent such as a cleansing oil, a cleansing foam, a soap, a body wash, and the like.
The cosmetic composition may further contain, in addition to the n-methylcytosine, a lipid, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, 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 cosmetics Such as, for example, cosmetics, cosmetics, pharmaceuticals, cosmetics, pharmaceuticals,
The cosmetic formulation may contain a relatively high concentration of n-methylcytosine in the case of wash-off type cosmetics such as make-up remover, detergent, etc. in which the active ingredient remains on the skin in a short period of time. On the other hand, in the case of leave-on type cosmetics such as lotion, cream, essence and the like in which the active ingredient remains on the skin for a long period of time, the concentration of the n-methylcitycin which is lower than that of the wash- It may be included. In one embodiment of the present invention, but not limited thereto, the composition comprises 0.0001 wt% to 10 wt% (preferably 0.0001 wt% to 1 wt%) of n-methylcytosine based on the total composition weight . When the composition of the present invention contains less than 0.0001 wt% of n-methylcytosine, sufficient skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement and skin barrier strengthening effect can not be expected, It is intended to prevent an undesired reaction such as allergy or a problem of skin safety.
Further, the composition for improving skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement and skin barrier enhancement of the present invention can be used for the preparation of external preparation for skin.
When the n-methylcytosine is used as an active ingredient of the external preparation for skin, it is further possible to use a lipid, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, , Surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, lipophilic or lipophilic active agents, And any other ingredients conventionally used in external preparations. The components can also be introduced in amounts commonly used in the field of dermatology.
When the n-methylcyclosine is provided as an external preparation for skin, it may have a formulation such as, but not limited to, ointments, patches, gels, creams or sprays.
The compositions of the present invention may also be used for the manufacture of pharmaceutical formulations. Thus, the composition of the present invention may comprise a pharmaceutically acceptable salt of said n-methylcytosine. The pharmaceutically acceptable salt of n-methylcytosine may be an acid addition salt formed using an organic acid or an inorganic acid. The organic acid may be, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid The organic acid may be selected from the group consisting of hydrochloric acid, hydrobromic acid, hydrochloric acid, hydrobromic acid, hydrobromic acid, hydrobromic acid, hydrobromic acid, hydrobromic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, , Benzenesulfonic acid, p-toluenesulfonic acid and methanesulfonic acid-based salts, and the inorganic acid includes, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid-based salts. Preferably in the hydrochloride or acetate form, more preferably in the hydrochloride form.
The above-mentioned acid addition salts may be prepared by a) directly mixing the n-methylcytosine and the acid, or b) dissolving and mixing one of them in a solvent or a water solvent, or c) Are placed in an acid in the sub-solvent and mixed therewith.
In addition to the above, additionally saltable forms include, but are not limited to, the salts of gabapentin, pregabalin, nicotinate, adipate, hemimarate, cysteine, acetylcysteine, methionine, arginine, Aspartate and the like.
The composition may further contain one or more active ingredients exhibiting the same or similar functions. For example, it may contain known skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening and skin tone improving ingredients. Wrinkle improvement, skin elasticity enhancement, skin whitening, and skin tone improving composition, the effect of the composition of the present invention on skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening and skin texture improving effect can be further enhanced. When the above ingredients are added, skin safety, easiness of formulation, and stability of effective ingredients can be considered according to the combined use. In one embodiment of the invention, the composition is a skin regeneration component known in the art, including retinoic acid, TGF, animal placenta-derived protein, betulinic acid and chlorella extract, (Hydroquinone), vitamin-C (L-ascorbic acid) and derivatives thereof, and various plant extracts, such as kojic acid and arbutin, Or may further comprise two or more components. The additional ingredient may be included in an amount of 0.0001% to 10% by weight based on the total weight of the composition, and the content range may be adjusted according to requirements such as skin safety, ease of formulation of n-methylcytosine, and the like .
In addition, the composition of the present invention may further comprise a pharmaceutically acceptable carrier.
Pharmaceutically acceptable carriers may contain a variety of ingredients such as buffer, injectable sterile water, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates, and the like.
The composition of the present invention can be administered orally or parenterally, and can be administered in the form of a general pharmaceutical preparation, for example, various forms of oral and parenteral administration at the time of clinical administration. In the case of formulation, a filler, , A binder, a wetting agent, a disintegrant, a surfactant, and the like.
Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin, and the like.
In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of liquid formulations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are commonly used simple diluents.
Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.
In the present invention, the term "effective amount" refers to an amount effective for promoting regeneration of damaged skin, improving wrinkles, improving elasticity, exhibiting a whitening effect, inhibiting or alleviating oxidation of cells, Means the amount of a compound capable of improving dry skin such as atopy, improving moisturizing effect, improving skin barrier, or improving skin texture. When the composition of the present invention contains an effective amount of the above-mentioned n-methylcytosine, it is possible to provide desirable skin regeneration effect, wrinkle improving effect, elasticity improving effect, skin whitening effect, skin texture improving effect and skin barrier strengthening effect. The effective amount of the n-methylcytosine contained in the composition of the present invention will vary depending on the form in which the composition is commercialized, the manner in which the compound is applied to the skin, and the time on the skin. For example, when the composition is made into a pharmaceutical formulation, it may contain the n-methylcytosine at a higher concentration than when it is commercialized as cosmetics that are routinely applied to the skin. Accordingly, the daily dose is 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, more preferably 50 to 60 mg / kg, based on the amount of the n-methylcytine, 6 times a day.
The 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.
Further, the composition for skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement and skin barrier enhancement of the present invention can be used for manufacturing food formulations.
The food formulation refers to food prepared by adding the n-methylcytosine to a food material such as beverage, tea, spice, gum, confectionery, or the like, or encapsulating, pulverizing, or suspending.
Since the food formulation can be taken on a daily basis, it is very useful because it can expect high skin regeneration, wrinkle improvement, elasticity enhancement, skin whitening, skin texture improvement and skin barrier enhancement effect.
When n-methylcytosine is used as a food additive, the n-methylcytosine may be added as it is or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, when the food or beverage is produced. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, 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 exceeding the above range .
There is no particular limitation on the kind of the food. Examples of the food to which the above substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.
When the food is a beverage, various flavors or natural carbohydrates may be added as an additional ingredient such as a normal drink. The above-mentioned natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio 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 food formulations may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, And the like. Other food formulations may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, 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 stimulating stem cell activity comprising the n-methylcytosine as an active ingredient.
The term 'stem cell' in the present invention is a cell capable of cell division by itself and capable of differentiating into a very specific type of specific cell type. The type of such stem cells is not particularly limited, and in one embodiment, the stem cells may be dermal stem cells. The term 'dermal stem cells' refers to stem cells that can be differentiated into cells constituting the skin (epidermis, dermis and subcutaneous fat layer). The cells that make up the skin include keratinocytes, melanocytes, and fibroblasts (mainly responsible for biosynthesis of collagen and elastin) present in the epidermis.
The kind of the skin stem cell is not particularly limited. The dermal stem cells used in the present invention can be used irrespective of where they originate from. For example, dermal stem cells may be obtained from a known source of dermal stem cells, e. G., From the basal layer of hair follicles or epidermis, and the animal to be harvested may be a mammal. In one embodiment, the mammal may include, but is not limited to, a human, a mouse, a rat, a guinea pig, a rabbit, a monkey, a pig, a horse, a cattle, a sheep, Preferably the mammal can be human. Such methods of obtaining dermal stem cells from dermal stem cell sources are well known in the art.
In the present invention, 'stem cell activity promoting effect' refers to a stem cell proliferation promoting effect and / or a stem cell maintaining effect. The stem cell maintenance is effected by promoting the expression of p63, which is a maintenance marker of dermal stem cells, as an intracellular transcription factor when treating n-methylcytosine to stem cells.
In addition, the composition for promoting stem cell activity of the present invention can be used for the production of cosmetic formulations. These cosmetic formulations are the same as those described in the description of cosmetic formulations including n-methylcytosine.
Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.
Reference Example 1: N- Methylcytisine Material Information
[Chemical Formula 1]
CAS No .: 486-86-2
Where to buy: Sichuan Hongjie Import & Export CO., LTD
Reference Example 2: Serum-free Under medium conditions Of dermal stem cells culture
Human epidermal stem cells purchased from Cellntec were added to 48-well plates (6 × 10 3 cells / well), supplemented with BPE (bovine pituitary extract) similar to fetal bovine serum And cultured for 24 hours at 5% CO 2 and 37 ° C using CNT-57 medium (Cellntec). Thereafter, the culture medium was removed through a suction tube, and the medium was removed using a PBS solution (GibcoBRL). To the CNT-57 medium without BPE, n-methylcytosine 0.1 μg / mL And cultured for 72 hours at 5% CO 2 and 37 ° C.
Experimental Example One: CCK -8 Through evaluation method Dermal stem cells Proliferation promoting effect
The skin stem cells cultured in Reference Example 2 were evaluated for CCK-8 (Cell counting kit-8). The CCK-8 evaluation is an indirect method of measuring the density of living cells by measuring the absorbance of Formazan, a dehydrogenase in the intracellular electron transport system, produced by decomposition of tetrazolium salt.
Cells were treated spectrophotometrically by treating CCK-8 solution (treated with 1/10 of the medium volume) at 37 ° C for 2 hours and then measuring the absorbance at 450 nm. The proliferation rate (%) of n-methylcytosine-induced dermal stem cell proliferation was calculated from the measured absorbance value in accordance with the following equation (1) with respect to the control group (CNT-57 medium supplemented with BPE) .
[Equation 1]
Growth rate (%) = (absorbance of sample-treated group / absorbance of control group) x 100
As shown in Table 1, the dermal stem cells under the medium conditions treated with n-methylcyclin exhibited an excellent proliferation promoting effect.
Experimental Example 2: Of dermal stem cells Stem cell ( stemness ) Confirm maintenance effect
The expression level of p63 was evaluated in order to confirm the stem cell maintenance effect of the dermal stem cells cultured in Reference Example 2 above. p63 is an intracellular transcription factor and is well known as a maintenance marker for dermal stem cells. Cells were isolated and cDNA was synthesized. Then, real-PCR was performed with Taqman staining solution to measure the expression level of p63. The concentration of RNA in this experiment was normalized to S16 ribosomal RNA.
The experimental results are shown in Table 2 below. In Table 2 below, the numerical value of the increase in p63 is a multiple of the control group (CNT-57 medium supplemented with BPE).
As shown in Table 2, the medium conditions in which n-methylcytosine was treated promoted the expression of p63 in dermal stem cells, thus exhibiting excellent stem cell maintenance effect.
Example 1: Promoting collagen synthesis
n-Methylcytosine was added to the culture medium of human-derived fibroblasts to examine the promoting effect of collagen synthesis at the cellular level. The biocompatible collagen was quantitated using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immunoassay Kit).
was added to the culture medium of human fibroblasts (7 x 10 4 cells / cm 2 ) together with vitamin C and a control group (no addition) so that the final concentrations of n-methylcystine were 0.05 ㎍ / mL and 0.1 ㎍ / After culturing for 1 day, the culture broth was taken and the degree of collagen biosynthesis at each concentration was measured at 450 nm using a spectrophotometer with PICP EIA Kit. The collagen biosynthesis performance was calculated by the relative performance relative to the control group and the results are summarized in Table 3 below.
As shown in Table 3, n-methylcytosine has excellent collagen synthesis ability against human-derived fibroblasts, and has superior collagen synthesis effect at a lower concentration than vitamin C, which is generally known to have collagen synthesis ability .
Example 2: Whitening effect - Confirmation of melanin formation inhibitory effect
n-methylcytosine was added to the culture of mouse melanoma cells (B-16 mouse melanoma cell) to test the whitening effect at the cellular level (Lotan R., Lotan D. Cancer Res. 40: 3345-3350, 1980 ). At this time, the toxicity of the melanoma cells of the rats before the experiment was evaluated, and the whitening evaluation was performed by selecting the concentration without toxicity.
Methylcytosine was added to the medium to a final concentration of 0.05 μg / mL and 0.1 μg / mL, and the control group, arbutin, was added to the medium to a concentration of 200 μg / mL, and treated with B-16 melanoma cells to obtain 3 Lt; / RTI >
Cells were then trypsinized, detached from the culture, centrifuged, and extracted with melanin. The removed cells were incubated with 1 mL of sodium hydroxide solution (1N), boiled for 10 minutes to dissolve melanin, and the absorbance was measured at 400 nm using a spectrophotometer to determine the amount of melanin produced.
The amount of melanin was measured by an absorbance of 10 6 cells per unit cell, and the amount of melanin produced relative to the control group was calculated as inhibition (%). The results are summarized in Table 4 below.
As shown in Table 4, it can be seen that n-methylcytosine has remarkably superior melanin production inhibitory effect on melanoma cells of the rat cultured as compared with arbutin, which is a known whitening substance.
Example 3: To improve the turn-over of the skin About Efficacy
About the nutritional cream of the following formulation example 2, the effect of the improvement of the exfoliation of the skin in the healthy twenties to 50 women was tested as follows.
Twenty of 20 women aged 20 to 50 years were given a 1.5% solution of DHA (Dihydroxyacetone, Sigma Aldrich, USA) for 8 hours, followed by deposition. The preparation cream was applied twice daily to the test site. After 4 days of application, 8 days after application, 12 days after application, photographs were taken using instrument Chromameter CR-400 (Minolta, Japan) and skin lightness of DSLR and DSLR. The measured values were evaluated by three mean values excluding the maximum value and the minimum value. The higher the improvement in the skin brightness of the deposition site, the better the improvement of the exfoliation. The results are shown in Table 5 below.
As shown in Table 5, when the nutritional cream according to the present invention was used, it was found that the exfoliation of the skin was improved.
Formulation example 1: Preparation of pharmaceutical preparations
1. Preparation of tablets
0.2 mg of n-methylcytosine
100 mg of corn starch
100 mg of milk
2 mg of magnesium stearate
After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.
Formulation example 2: Manufacture of cosmetics
1. Manufacture of nutritional cream
A nutritional cream containing n-methylcytosine as an active ingredient was prepared according to a conventional method, as shown below.
0.2% by weight of n-
Beta-1,3-glucan 5.0 wt%
Wax 10.0 wt%
Polysorbate 60 1.5 wt%
≪ tb > < tb > < tb >
0.5% by weight of sorbitan sesquioleate
Liquid paraffin 10.0 wt%
Squalane 5.0 wt%
Caprylic / capric triglyceride 5.0 wt%
Glycerin 5.0 wt%
3.0% by weight of butylene glycol
3.0% by weight of propylene glycol
0.2% by weight triethanolamine
Preservative 0.05 wt%
0.05% by weight of pigment
0.05% by weight fragrance
Purified water to 100%
Formulation example 3: Preparation of external preparation for skin
1. Manufacture of ointment
An ointment containing n-methylcytosine as an active ingredient was prepared according to a conventional method as follows.
0.5% by weight of n-
Beta-1,3-glucan 10.0 wt%
Wax 10.0 wt%
Polysorbate 60 5.0 wt%
≪ tb > < tb > < tb >
0.5% by weight of sorbitan sesquioleate
Vaseline 5.0 wt%
Liquid paraffin 10.0 wt%
Squalane 5.0 wt%
SHARE BUTTER 3.0 wt%
Caprylic / capric triglyceride 5.0 wt%
Glycerin 10.0 wt%
Propylene glycol 10.2 wt%
0.2% by weight triethanolamine
Preservative 0.05 wt%
0.05% by weight of pigment
0.05% by weight fragrance
Purified water to 100%
Claims (9)
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH08208424A (en) | 1994-12-20 | 1996-08-13 | Unilever Nv | Cosmetic composition containing betulinic acid |
| JPH08231370A (en) | 1995-02-23 | 1996-09-10 | Taiyo Kagaku Co Ltd | Skin cosmetic |
| JPH0940523A (en) | 1995-07-28 | 1997-02-10 | Ichimaru Pharcos Co Ltd | Fibroblast proliferation promoter containing water extract form chlorella |
| JPH1036283A (en) | 1996-07-18 | 1998-02-10 | Ichimaru Pharcos Co Ltd | Fibroblast proliferation promoting agent |
-
2015
- 2015-09-23 KR KR1020150134843A patent/KR20170035661A/en unknown
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH08208424A (en) | 1994-12-20 | 1996-08-13 | Unilever Nv | Cosmetic composition containing betulinic acid |
| JPH08231370A (en) | 1995-02-23 | 1996-09-10 | Taiyo Kagaku Co Ltd | Skin cosmetic |
| JPH0940523A (en) | 1995-07-28 | 1997-02-10 | Ichimaru Pharcos Co Ltd | Fibroblast proliferation promoter containing water extract form chlorella |
| JPH1036283A (en) | 1996-07-18 | 1998-02-10 | Ichimaru Pharcos Co Ltd | Fibroblast proliferation promoting agent |
Non-Patent Citations (4)
| Title |
|---|
| 1. Journal of the European Academy of Dermatology and Venereology. 12:103-114, 1999 |
| 2. The British journal of dermatology. 110: 129-138, 1984, Skin research and technology.5:189-194, 1999 |
| 3. Epidermal Stem Cells of the Skin, Cedric Blanpain, Elaine Fuchs, Annual Review of Cell and Developmental Biology, November 2006, Vol. 22, Pages 339-373 |
| 4. Human skin stem cells and the ageing process, Catherin Niemann, Stem Cell Aging and Regenerative Medicine, November 2008, Pages 986-997 |
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