KR102348696B1 - Cosmetic composition comprising teprenone - Google Patents

Abstract

The present invention relates to a composition for inducing differentiation of preadipocytes or stem cells into adipocytes, comprising Teprenone. In addition, the present invention provides a cosmetic composition comprising teprenone and an external preparation for skin, (a) culturing stem cells; and (b) adding a composition containing teprenone to the stem cells of step (a). Teprenone of the present invention promotes the differentiation of adipose tissue-derived stem cells into mature adipocytes by promoting the expression of C/ebp α, which is a key regulator of adipogenesis. It can present new possibilities to improve the aesthetic and health problems of people.

Description

Cosmetic composition comprising teprenone

The present invention relates to a composition for inducing differentiation of preadipocytes or stem cells into adipocytes, including teprenone. In addition, the present invention relates to a cosmetic composition containing teprenone and an external preparation for skin, and a method for inducing differentiation of stem cells into adipocytes using the composition containing teprenone.

The subcutaneous tissue is located between the dermis and the fascia and is a connective tissue including adipocytes, pre-adipocytes, adipose-derived stem cells, blood vessels, and neural networks that store body fat. Subcutaneous fat basically functions as energy storage, body temperature protection from external cold and heat, and buffering action. In addition, by secreting various hormones, growth factors and cytokines to regulate immunity, body temperature control, skin tissue growth and aging, it can play a role in improving the appearance of the skin through skin regeneration and suppression of wrinkle formation. . In addition, it provides elasticity and firmness to the skin, greatly affecting the shape of the individual's external state.

It is well known that such subcutaneous fat significantly reduces the overall volume of the tissue as it ages, and its function deteriorates. However, in modern times, adipose tissue tends to shrink even at a relatively young age due to the imbalance of influence caused by excessive diet. This causes problems such as reduction in skin elasticity and strength due to reduction in adipose tissue volume, as well as structural collapse due to loss of strength, promotion of aging due to deterioration of skin tissue and function as an endocrine organ that regulates systemic metabolism, reduced immunity and regeneration do.

In order to solve this problem, a method such as autologous fat grafting using an invasive method is currently being implemented. It is a method of collecting and moving fat from the abdominal and thigh areas, which are relatively rich in fat, and can partially improve the reduction of subcutaneous fat due to aging or skin sunken due to burns, surgery, and scars. However, it has to be relatively expensive, the engraftment rate of autologous adipose tissue after surgery is not constant, and it is an invasive method that has to use a surgical method. Accordingly, there is a growing demand for a non-invasive method capable of recovering the regression of the subcutaneous fat layer while compensating for these shortcomings.

Many recent studies have demonstrated that mesenchymal-type pluripotent stem cells exist in the subcutaneous fat. These stem cells are called adipose tissue-derived stem cells (ADSC), and according to environmental signals, they differentiate into mature adipocytes. In addition, it maintains the homeostasis of fat and skin tissue by secreting various hormones and cytokines. In addition, the engraftment of adipose tissue after autologous fat grafting is regulated by the content of adipose tissue-derived stem cells. That is, by inducing adipose tissue-derived stem cells to differentiate into subcutaneous fat, a non-invasive and safe fat regeneration with the same effect as autologous fat graft surgery can be expected.

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 removed 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. retains its function as

In addition, dry skin, 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, It is increasing due to various factors such as excessive bathing culture and skin aging, 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.

On the other hand, teprenone is a compound having a structure of the following formula (1), also called geranylgeranylacetone (geranylgeranylacetone), the chemical formula is C ₂₃ H ₃₈ O is a compound having a molecular weight of 330.55. Currently, Teprenone is mainly known for its use to improve gastric mucosal lesions (erosion, bleeding, redness, edema) or to improve acute gastritis and chronic gastritis.

[Formula 1]

Under this background, the present inventors found that Teprenone promotes the differentiation of adipose tissue-derived stem cells into mature adipocytes by promoting the expression of C/ebp α, which is a key regulator of adipogenesis, and found that Teprenone, which was previously unknown By revealing for the first time the function of regulating adipocyte differentiation, the present invention was completed.

It is an object of the present invention to provide a composition for inducing differentiation of preadipocytes or stem cells into adipocytes, comprising a teprenone compound or an acceptable salt thereof.

Another object of the present invention is to provide a cosmetic composition comprising a teprenone compound or a cosmetically acceptable salt thereof.

Another object of the present invention is to provide an external preparation for skin comprising a teprenone compound or an acceptable salt thereof.

Another object of the present invention comprises the steps of (a) culturing stem cells; and (b) adding the composition containing Teprenone to the stem cells of step (a).

The present invention provides a composition for inducing differentiation from preadipocytes or stem cells into adipocytes comprising a teprenone compound or an acceptable salt thereof to solve the above problems and achieve the object of the present invention. to provide.

In the present invention, "Teprenone (Teprenone)" is a compound represented by the structure of the following formula (1), the molecular formula is C ₂₃ H ₃₈ O, the molecular weight is a compound of 330.55. In addition to the above terms, Teprenone is also called geranylgeranylacetone.

[Formula 1]

The present invention is not particularly limited to the method for obtaining teprenone, and may be chemically synthesized by a method known in the art, or a commercially available material may be used.

The teprenone represented by Formula 1 of the present invention may exist in a non-solvated form as well as a solvated form. The teprenone of the present invention may exist in crystalline or amorphous form, and all such physical forms are included within the scope of the present invention.

In the composition according to the present invention, the content of teprenone may include 0.0001 to 50% by weight of teprenone relative to the total weight of the composition, and specifically 0.01 to 10% by weight of the cosmetic composition of the present invention relative to the total weight of the composition. It may be included in weight %. It exhibits excellent lipofilling, fat filling or skin regeneration effects within the above range, and at the same time has the advantage of stabilizing the formulation of the composition.

In the present invention, the term "progenitor cell" is referred to as a committed stem cell, and when a specific differentiation-forming expression of a cell (X) corresponding to a progeny is revealed, an undifferentiated parent cell that does not express the differentiation trait is a progenitor cell of X call it In addition, "adipocyte progenitor cell" refers to a progenitor cell whose differentiation into adipocyte expression has been revealed.

In the present invention, the term "stem cell" refers to a cell that exists in all multicellular organisms as undifferentiated cells of animals, and has the ability to reproduce indefinitely, maintain normal chromosomes, and differentiate into various cells. Since it was discovered by Ernest A. McCulloch and James E. Till of Ontario Cancer Institute in Canada in the 1960s, in particular, due to the totipotent potential of being able to differentiate into any cell of the tissues constituting more than 210 organs in the body, the human body known as the master cell of These stem cells are not limited thereto, but may be embryonic stem cells, adult stem cells, umbilical cord blood stem cells, mesenchymal stem cells, or human adipose tissue-derived stem cells, depending on the origin. More specifically, it may be a mesenchymal stem cell, or more specifically, it may be an adipose tissue-derived mesenchymal stem cell.

As used herein, the term "animal" includes not only humans and primates, but also domestic animals such as cattle, pigs, sheep, horses, dogs, mice, rats or cats, specifically humans.

As used herein, the term "embryonic stem cell (ESC)" is a stem cell derived from the inner cell mass (ICM) of the blastocyst stage, which is one of the early developmental stages of the embryo. About 4-5 days later, it has 50-150 cells, and these cells are stem cells with pluripotent differentiation ability that can divide into more stem cells or differentiate into tridermal cells. .

As used herein, the term "umbilical cord blood stem cells" refers to stem cells present in blood from the placenta and fetus' umbilical cord, and has a stronger proliferation than bone marrow-derived stem cells.

As used herein, the term "mesenchymal stem cells" refers to mesenchymal stem cells that are differentiated into mesenchymal tissues such as muscle, bone, and fat, which exist in the bone marrow and are adult stem cells having the ability to differentiate into cells such as nerves and skin. It refers to one of the cells (adult stem cell, AS). These adult stem cells are found in organs that repeat continuous cell replacement, and these organs include the epidermal layer of the skin, the lining of the small intestine, bone marrow, brain, tendons, and the like.

As used herein, the term "adipose tissue-derived stem cells" refers to cells with multipotency distributed in adipose tissue. They have the same level of differentiation potential as those of bone marrow mesenchymal stem cells, and it is reported that they are superior to bone marrow mesenchymal stem cells in maintaining colony forming ability and proliferative capacity in cell culture. Adipose tissue-derived stem cells increase the growth rate and cell cycle when antioxidants such as N-acetyl-L-cystein and ascorbic acid-2-phosphate are added and the calcium concentration is low. In addition, the proliferation of adipose tissue-derived stem cells can be stimulated by FGF-2 (fibroblast growth factor), and adipocyte-derived stem cells are vascular endothelial growth factor (VEGF), hepatocyte growth factor (hepatocyte growth factor). It is known to secrete potential growth factors such as factor, HGF) and insulin like growth factor (IGF). Unlike stem cells isolated from bone marrow or umbilical cord blood, adipose tissue-derived stem cells have the advantage that they can be collected repeatedly and do not cause pain to the patient when collected like bone marrow. It has the advantage of no rejection. In addition, it is easier to culture than bone marrow-derived mesenchymal stem cells, the amount of isolated mesenchymal stem cells is 100 to 1000 times higher than that of bone marrow, and self-renewal ability and multipotent ability are also known to be excellent.

As used herein, the term “differentiation into adipocytes” refers to the differentiation of preadipocytes or stem cells of an animal, specifically, a mammal into adipocytes. As used herein, the term “composition for inducing differentiation” refers to a composition capable of inducing a process in which cells in the initial stage have characteristics as individual tissues, and for the purpose of the present invention, the preadipocytes or stem cells are transformed into adipocytes. means a composition capable of inducing differentiation into Specifically, the composition for inducing differentiation may induce differentiation of adipocytes by promoting the expression of C/ebp α.

As used herein, the term "C/ebp α (CCAAT-enhancer 0 binding protein α)" is a protein belonging to C/EBPs (CCAAT-enhancer 0 binding proteins) and is a transcription factor. These proteins are known to promote the expression of specific genes by binding to their promoters. In the present invention, it can be used as an adipocyte differentiation marker to determine whether adipose tissue-derived stem cells are induced to differentiate into adipocytes by measuring the expression level of the C/ebp α.

In a specific embodiment of the present invention, it was confirmed that the differentiation of adipocytes occurred more effectively than when Teprenone was added and then the differentiation of adipose tissue-derived stem cells into adipocytes was induced, compared to the case where the adipocytes were not added. By promoting the expression of C/ebp α, a marker for measuring cell differentiation, it was confirmed that the adipose tissue-derived stem cells had an excellent effect on promoting adipocyte differentiation ( FIGS. 1 to 3 ). These results show that the composition containing the teprenone compound of the present invention has a function of regulating the differentiation of adipocytes, so it is a novel possibility to improve the aesthetic and health problems of modern people caused by volume reduction and functional deterioration due to adipose tissue regression. is a result that presents

As another embodiment, the present invention provides a cosmetic composition comprising a teprenone compound or a cosmetically acceptable salt thereof.

In addition, specifically, the present invention may provide a cosmetic composition for lipofilling or fat filling, or skin regeneration comprising a Teprenone compound or a cosmetically acceptable salt thereof.

In the present invention, the term "Teprenone" is the same as described above.

In the present invention, "lipofilling (lipofilling) or fat filling" means to relieve, improve, or fill the degree of reduction in the volume of adipose tissue due to the regression of adipose tissue in the skin. It means that the volume is improved.

As used herein, the term "skin regeneration" means to allow severely damaged skin to recover its original function, but is not limited thereto, but skin regeneration relieves the degree of sagging or sagging of the skin, or filling the skin with fat or fat. It may include maintaining skin elasticity by filling fat with structural breakdown due to decreased skin elasticity and strength due to reduction in tissue volume. In addition, it may include inhibiting or inhibiting the generation of wrinkles on the skin, or alleviating the wrinkles already generated.

In one embodiment of the present invention, the teprenone compound promotes the expression of C/ebp α, which is a key regulator of adipogenesis, so that adipose tissue-derived stem cells, more specifically adipose tissue-derived mesenchymal stem cells, are converted into mature adipocytes. It was confirmed that differentiation was induced. These results suggest that by inducing the differentiation of stem cells into mature adipocytes, it is possible to alleviate, improve, or fill the decreased volume of skin adipose tissue. Therefore, the cosmetic composition containing the teprenone compound of the present invention This indicates that it can be used for lipofilling or fat filling.

In addition, the above results suggest that by alleviating or improving the volume of regressed adipose tissue, it is possible to promote or improve skin elasticity and wrinkles, and furthermore, to help damaged skin recover its original function. This suggests that the cosmetic composition comprising the teprenone compound of the present invention can be used for skin regeneration.

The cosmetic composition is a solution, external ointment, cream, foam, nutrient lotion, softening lotion, pack, soft water, emulsion, makeup base, essence, soap, liquid detergent, bath agent, sunscreen cream, sun oil, suspension, emulsion, It may be prepared in a formulation selected from the group consisting of paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch and spray, but is not limited thereto.

In addition, the cosmetic composition may further include one or more cosmetically acceptable carriers to be formulated in general skin cosmetics, and as common ingredients, for example, oil, water, surfactant, humectant, lower alcohol, thickener, A chelating agent, a colorant, a preservative, a fragrance, etc. may be appropriately blended, but the present invention is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition varies depending on the formulation.

When the formulation of the cosmetic composition is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide as a carrier component or mixtures thereof may be used.

When the formulation of the cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or a mixture thereof may be used as a carrier component, and in particular, in the case of a spray, additionally propellants such as chlorofluorohydrocarbons, propane/butane or dimethyl ether.

When the formulation of the cosmetic composition is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol , 1,3-butylglycol oil can be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty ester, fatty acid ester of polyethylene glycol or sorbitan may be used. can

When the formulation of the cosmetic composition is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the formulation of the cosmetic composition is soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolsate, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oil, glycerol, sugar, etc. are used as carrier components can be

As another specific embodiment, the composition comprising the teprenone compound of the present invention may be used as a food composition.

The food composition may be for lipofilling or fat filling containing a Teprenone compound or a pharmaceutically acceptable salt thereof, or for skin regeneration.

The food composition may be used in the form of a health functional food, but is not limited thereto.

Teprenone or a pharmaceutically acceptable salt thereof contained in the food composition may be included in the form of animals and plants containing Teprenone, an extract thereof, a fraction thereof, or a processed product thereof. In addition, the composition may include a food pharmaceutically acceptable food supplement additive in addition to the active ingredient.

In the present invention, "food supplement additive" refers to a component that can be added to food as an auxiliary, added to the manufacture of health functional food of each formulation, and can be appropriately selected and used by those skilled in the art. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of food supplement additive of the present invention.

The food composition of the present invention may include a health functional food. In the present invention, "health functional food" refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful for the human body. Here, "functional" means to obtain useful effects for health purposes, such as regulating nutrients or physiological actions with respect to the structure and function of the human body. The health functional food can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art. In addition, the dosage form of the health functional food may be manufactured without limitation as long as it is a dosage form recognized as a health functional food. The food composition of the present invention can be prepared in various forms, and unlike general drugs, it has the advantage that there are no side effects that may occur during long-term administration of the drug using food as a raw material, and excellent portability, the present invention Health functional food of lipofilling (lipofilling), fat filling, or can be consumed as a supplement to enhance the skin regeneration improvement effect.

There is no limitation in the form that the health functional food of the present invention can take, and it may include any food in a conventional sense, and may be used interchangeably with terms known in the art, such as functional food. In addition, the health functional food of the present invention can be prepared by mixing known additives with other suitable auxiliary ingredients that may be included in food according to the selection of those skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and There are vitamin complexes, and the like, and it can be prepared by adding the compound represented by Formula 1 according to the present invention as a main component to juice, tea, jelly, juice, and the like. It also includes food used as feed for animals.

As another specific embodiment, the composition including the teprenone of the present invention can be used as a quasi-drug composition.

In addition, specifically, the quasi-drug composition may provide a quasi-drug composition for lipofilling or fat filling, or skin regeneration comprising the teprenone or a pharmaceutically acceptable salt thereof.

The quasi-drug composition may further include a pharmaceutically acceptable carrier, excipient or diluent, if necessary, in addition to the above ingredients. The pharmaceutically acceptable carrier, excipient or diluent is not limited as long as it does not impair the effects of the present invention, and includes, for example, fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, fragrances, preservatives, etc. may include

Representative examples of pharmaceutically acceptable carriers, excipients or diluents of the quasi-drug composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, maltitol, starch, gelatin, glycerin, acacia gum, alginate, calcium phosphate, Calcium carbonate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxy benzoate, talc, magnesium stearate, mineral oil, propylene glycol, polyethylene glycol, vegetable oils, injectable esters, witepsol, macrogol, tween 61, cacao butter, laurage, and the like.

In addition, when Teprenone of the quasi-drug composition is used as a quasi-drug, it may contain one or more additional active ingredients having the same or similar function. For example, it may include known lipofilling or fat filling, or skin regeneration ingredients. When an additional lipofilling or fat filling, or skin regeneration component is included, the lipofilling or fat filling, or skin regeneration improvement effect of the quasi-drug composition of the present invention may be further increased. When adding the above ingredients, skin safety according to complex use, ease of formulation, and stability of active ingredients can be considered.

The quasi-drug composition of the present invention may exemplify, but is not limited to, disinfectant cleaner, shower foam, ointment, wet tissue, coating agent, etc. It may be appropriately selected from conventional techniques.

In addition, the teprenone of the present invention can be used in a method of lipofilling or fat filling, or skin regeneration, including the step of applying to the skin of an individual. The subject includes, without limitation, mammals including rats, livestock, humans, and the like.

As another embodiment, the present invention provides an external preparation for skin comprising a teprenone compound or an acceptable salt thereof.

In addition, specifically, the present invention may provide an external preparation for lipofilling or fat filling or skin regeneration including a Teprenone compound or an acceptable salt thereof.

In the present invention, the terms "Teprenone", "for lipofilling or fat filling", and "for skin regeneration" are the same as described above.

As used herein, the term "external preparation" is a preparation provided for external use, and includes external acid preparations, external tablets, external solutions, ointments, warning preparations, suppositories, and the like. External preparation means dispersing zincized starch, etc., erosions and ulcers of the skin mucous membrane, external tablet means dissolving or vaginal tablet, and external preparation is a liquid containing water, ethanol, oil, etc. as a solvent It is a preparation, and is used for brushing teeth, compresses, washing, eye drops, and nasal drops. In addition, an ointment is a semi-solid formulation of suitable roughness applied to the skin, a warning is a solid at room temperature, a skin application that softens at body temperature, and a suppository is a formulation applied to the anus, vagina, and urethra.

The “external preparation for skin” refers to a preparation that acts externally on the skin among external preparations, and in the present invention, it refers to an external preparation for skin containing the Teprenone compound or an acceptable salt thereof as an active ingredient. Specifically, for external skin preparations, creams, gels, patches, sprays, ointments, warnings, lotions, liniments, pastas, or cataplasmas can be prepared and used as pharmaceuticals or quasi-drugs in the form of external preparations for the skin, but is not limited thereto. does not

As another one embodiment, the present invention comprises the steps of (a) culturing stem cells; And (b) it relates to a method for inducing differentiation into adipocytes from stem cells, comprising adding a composition comprising a teprenone compound to the stem cells of step (a).

In the present invention, the terms "Teprenone", "stem cells" and "differentiation induction" are the same as described above.

The step (a) is a step of culturing undifferentiated cells, which are stem cells, and can be cultured according to a conventional culture method known in the art. , more specifically, may be adipose tissue-derived mesenchymal stem cells.

The step (b) is a step of inducing differentiation into adipocytes by treating the stem cells cultured in step (a) with the composition containing the eteprenone of the present invention. Cytokines added to the culture medium to induce differentiation can be easily selected by those skilled in the art from cytokines for adipocyte differentiation known in the art, and can be used by adding an effective concentration for inducing differentiation to the medium, It is not limited thereto.

Specifically, as described above, the method for inducing adipocyte differentiation of the present invention can promote the expression of C/ebpα, and the adipocytes differentiated from the stem cells produced in the present invention can be used as cosmetics, food, quasi-drugs, or It may be used as an external preparation for skin, and in particular, the cosmetic, food, quasi-drug, or external preparation for skin may be used for lipofilling or fat filling, or skin regeneration.

In addition, it can be used as a cell composition, such as a cell therapeutic agent for the treatment of rare incurable diseases and diseases related to adipocyte transplantation.

As described above, the teprenone of the present invention promotes the differentiation of adipose tissue-derived stem cells into mature adipocytes by promoting the expression of C/ebp α, which is a key regulator of adipogenesis, thereby reducing the volume due to adipose tissue shrinkage and It can present a new possibility to improve the aesthetic and health problems of modern people caused by functional deterioration.

Figure 1 is indomethacin (Indomethacin), dexamethasone (Dexamethason), isobutyl methyl xanthine (IBX), insulin (Insulin) containing adipocyte differentiation conditions, adipose tissue-derived mesenchymal stem cells fat After inducing cell differentiation, fat was stained with Oil Red O staining reagent and observed under a microscope. The non-differentiation group was a control group under culture conditions that did not induce differentiation, the untreated control group was a control group with only adipocyte differentiation medium added, a positive control group was obtained by adding Rosiglitazone to the adipocyte differentiation medium, and the experimental group was an adipocyte differentiation medium. It was carried out by adding tefrenone to it.
Figure 2 is a graph showing the relative adipogenic differentiation level compared to the untreated control group after the adipose tissue-derived mesenchymal stem cells were treated with an adipocyte differentiation medium containing teprenone.
3 is a graph showing the expression level of C/ebp α, a genetic marker for adipocyte differentiation, compared to a control group after adipose tissue-derived mesenchymal stem cells were treated with an adipocyte differentiation medium containing Teprenone.

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Reference example One : tefrenone ( teprenone ) substance information

[Formula 1]

Substance name: teprenone

CAS No. : 6809-52-5

Molecular Formula: C ₂₃ H ₃₈ O

Molecular Weight: 330.55

Where to buy: Sigma-Aldrich

Example 1: Confirmation of the efficacy of promoting differentiation of human adipose tissue-derived stem cells into adipocytes

Human adipose-derived stem cells (hADSCs) were purchased from Lonza (Lonza, Inc. Walkersville, MD, USA) and cultured according to Lonza's instructions. Adipocyte differentiation was achieved using the method recommended by Lonza. Specifically, 0.5 μg/ml of insulin, 0.5 μM of dexamethasone (DEXA), 0.25 mM of isobutyl methyl xanthine (IBMX), and 50 μM of indomethacin were added to the adipose tissue-derived stem cell culture medium and cultured. Thus, stem cells were differentiated into adipocytes, and 10 ppm of teprenone was added to the medium under the same conditions to induce adipocyte differentiation. As a positive control, 0.5 μM of Rosiglitazone (ROSI) was used and treated with IDX medium. 14 days after the initiation of adipocyte differentiation of adipose tissue-derived stem cells, the cell culture medium was removed, the cells were washed with 10 ml of Phosphate Buffered Saline (PBS), and then the cells were cultured using 10% formalin solution. was fixed for a minute. The fixed cells were washed once with a 60% isopropanol solution, and the lipids in the adipocytes were stained with a staining solution in which Oil Red O was dissolved in 60% isopropanol for 10 minutes. Then, it was observed under a microscope, and the results are shown in FIG. 1 .

As a result, as can be seen in FIG. 1 , when Teprenone was added and then adipogenic differentiation of adipose tissue-derived stem cells was induced, compared to the case of inducing adipocyte differentiation of adipose tissue-derived stem cells under the conditions of no addition. It was confirmed that the differentiation of adipocytes occurred more effectively (FIG. 1).

These results indicate that the composition containing teprenone has a lipolysis use by promoting the differentiation of adipose tissue-derived stem cells into adipocytes, and furthermore, the composition containing teprenone is used for lipofilling or fat filling, or skin This suggests that it can be used for recycling purposes.

Example 2: Quantitative confirmation of adipocyte differentiation promoting effect of human adipose tissue-derived stem cells

After inducing differentiation of adipocytes from adipose tissue-derived stem cells in the same manner as in Example 1, adipocytes were stained with Nile Red reagent (AdipoRed™, Lonza). The staining method followed the method recommended by Lonza. Fluorescence of the stained adipocytes was measured at 485 nm/575 nm of 500 nm, and the results are shown in Table 1 and FIG. 2 .

As a result, as can be seen in Tables 1 and 2 above, as a result of treating adipose tissue-derived mesenchymal stem cells with teprenone, differentiation of adipose tissue-derived stem cells into adipocytes was induced under the condition in which Teprenone was not added. It was confirmed that the differentiation of adipocytes occurred more effectively than in one case (Table 1 and FIG. 2).

These results also indicate that the composition containing teprenone has a lipolysis application by promoting the differentiation of adipose tissue-derived stem cells into adipocytes. This suggests that it can be used for skin regeneration purposes.

Example 3: Confirmation of adipocyte-specific gene expression level in human adipose tissue-derived stem cells

After inducing differentiation of adipocytes from adipose tissue-derived stem cells in the same manner as in Example 1, the expression level of C/ebp α, a marker of adipocyte differentiation, in order to confirm the degree of adipocyte differentiation of adipose tissue-derived stem cells. was evaluated. Cell RNA was isolated to synthesize cDNA, and then, real-time-PCR was performed with Taqman staining solution to measure the expression level of C/ebp α. In this experiment, the RNA concentration was normalized to S16 ribosomal RNA, and the results are shown in Table 2 and FIG. 3 .

As a result, as can be seen in Table 2, the medium condition treated with Teprenone promoted the expression of C/ebp α, and the effect of promoting differentiation of adipose tissue-derived stem cells into adipocytes was greater than that of the case where Teprenone was not added. It was confirmed that it was excellent (Table 2 and FIG. 3).

These results indicate that the composition containing teprenone has a lipolysis use by promoting the differentiation of adipose tissue-derived stem cells into adipocytes, and furthermore, the composition containing teprenone is used for lipofilling or fat filling, or skin This suggests that it can be used for recycling purposes.

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

Claims (17)

A composition for inducing differentiation into adipocytes from preadipocytes or stem cells, comprising a teprenone compound or an acceptable salt thereof.
The composition of claim 1, wherein the stem cells are mesenchymal stem cells (MSCs).
The composition of claim 1, wherein the stem cells are human adipose tissue-derived stem cells.
The composition of claim 1, wherein the composition promotes the expression of C/ebp α.
A cosmetic composition for inducing differentiation of preadipocytes or stem cells into adipocytes, comprising a teprenone compound or a cosmetically acceptable salt thereof.
The cosmetic composition according to claim 5, wherein the composition has at least one of a lipofilling or fat filling effect, and a skin regeneration effect.
delete delete The cosmetic composition of claim 5, wherein the stem cells are mesenchymal stem cells (MSCs).
The cosmetic composition of claim 5, wherein the stem cells are human adipose tissue-derived stem cells.
According to claim 5, wherein the cosmetic composition is a solution, external ointment, cream, foam, nourishing lotion, softening lotion, pack, soft water, emulsion, makeup base, essence, soap, liquid detergent, bath agent, sunscreen cream, sun oil , suspension, emulsion, paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch and spray. , cosmetic composition.
A skin external preparation for inducing differentiation of preadipocytes or stem cells into adipocytes, comprising a teprenone compound or an acceptable salt thereof.
[Claim 13] The topical preparation for skin according to claim 12, wherein the external preparation for skin has at least one of a lipofilling or fat filling effect, and a skin regeneration effect.
delete (a) culturing the stem cells; and
(b) A method for inducing differentiation from stem cells into adipocytes, comprising adding the composition of claim 1 to the stem cells of step (a).
The method of claim 15, wherein the stem cells are mesenchymal stem cells.
The method of claim 15, wherein the stem cells are derived from human adipose tissue.

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