KR102409079B1 - Pharmaceutical composition for prevention or treatment of obesity comprising the extracts of Schizandra chinensis as active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating obesity containing an extract of Schizandra chinensis as an active ingredient, wherein the extract of Schisandra chinensis is UCP-1 (Uncoupling protein-1), a gene involved in energy consumption in white adipocytes. , because it increases the expression of PGC-1α and PRDM16, the composition of the present invention can be used as a brown fat inducer or anti-obesity agent, a pharmaceutical composition for inducing brown adipocytes, a health functional food, a cosmetic composition, and a pharmaceutical composition for external application to the skin .

Description

A composition for preventing and treating obesity comprising the extract of Schisandra as an active ingredient {Pharmaceutical composition for prevention or treatment of obesity comprising the extracts of Schizandra chinensis as active ingredient}

The present invention is Schisandra chinensis ) It relates to a pharmaceutical composition for preventing and treating obesity or for inducing brown adipocytes, a health functional food, a cosmetic composition, and a pharmaceutical composition for external use on the skin containing the extract as an active ingredient.

With the recent economic growth and changes in lifestyle, there have been many changes in eating habits. In particular, in the case of busy modern people, excessive weight and obesity are increasing due to a high-calorie diet such as fast food and a small amount of exercise. Overweight means a body mass index (BMI, a measure of obesity divided by weight squared) of 25 or more and less than 30, and obesity means a body mass index of 30 or more. Two-thirds of American adults are overweight or obese, and excess weight increases blood pressure and cholesterol levels, increasing the incidence of various adult diseases such as heart disease. The effect of obesity on various diseases is very serious, and it is known that obesity is the cause of 80% of diabetic patients and 21% of heart disease worldwide. In addition, various cancers such as uterine cancer, kidney cancer, and breast cancer are also directly or indirectly related to obesity, and in the case of obesity or overweight, the incidence of sleep apnea, osteoarthritis, and cholelithiasis is higher than that of normal people (Stein CJ, Colditz GA. The epidemic of obesity.J Clin Endocrinol Metab 2004;89: 2522-2525. Kopelman PG. Obesity as a medical problem.Nature 2000 Apr 6;404(6778):635-43.). Obesity is not a disease caused by a single cause, but is difficult to treat with any one method because it is caused by a combination of several factors such as genetics, environment-social, mental, etc. Currently, methods for treating obesity can be divided into lifestyle correction methods such as diet therapy, exercise therapy, and behavior therapy, as well as drug therapy and surgical treatment. Active efforts to correct lifestyle should be preceded by drug or surgical treatment, but it is not easy to correct lifestyle, and there is a limit to the weight that can be reduced only by lifestyle modification. Therefore, in many cases, drug treatment is necessary along with lifestyle modification.

Although many anti-obesity agents are being developed every year for the drug treatment of obesity, there are not many currently available anti-obesity drugs, and most are limited to agents that suppress digestion or appetite. In the case of agents that control digestion or appetite, they are classified as psychotropic drugs because of their habit, and digestive inhibitors exhibit side effects such as diarrhea and constipation. The representative obesity treatment drugs approved for long-term use by the U.S. FDA until 2010 are norepinephrine, sibutramine (Reductil) that inhibits the reuptake of serotonin, and secretion from the pancreas and digestive system There was orlistat (Xenical), which showed an effect by inhibiting lipase (Yanovski SZ, Yanovski JA. Obesity. N Engl J Med 2002; 346: 591-602.). However, sibutramine was withdrawn from the United States and Korea in October 2010 because side effects such as elevated blood pressure, insomnia, dry mouth, and dizziness are common, and there is a risk of cardiovascular symptoms such as myocardial infarction and stroke. In addition, orlistat has common side effects such as diarrhea, fatty stools, and loss of money, and its use is limited because the effect of the drug is not clear when the intake of fat is lower than that of Westerners such as Koreans (Sangman Kim. A study on Orlistat. Journal of the Korean Society for Obesity) 1998; 7(4): 287-92.). Therefore, there is a need for anti-obesity and therapeutic agents to replace obesity because there is still no anti-obesity drug with good anti-obesity effect and safety for long-term use. In general, fat stored in adipocytes is used as an important energy source in the body, but obesity progresses due to excessive caloric intake and inefficient consumption of caloric energy. At this time, along with the increase in the number of adipocytes, a large amount of triglyceride synthesis is induced by excessive differentiation of adipocytes, accompanied by morphological changes including an increase in the size of adipocytes and changes in the expression of various genes. The increase in adipose tissue is achieved by an increase in the number of adipocytes and an increase in the size of adipocytes. In particular, although the size of adipocytes is generally possible with diet control, the process of new precursor adipocytes differentiated into adipocytes is not effective with diet control. It is important to control

Mammalian adipocytes are classified as either white adipocytes, which store energy, or brown adipocytes, which consume energy. Brown adipocytes express uncoupling protein-1 (UCP1), which converts biochemical energy into heat by unconjugated ATP production from a mitochondrial proton gradient (Cannon et al., 2004, Physiol Rev). 84:277-359). This heat plays a role in maintaining body temperature in a cold environment or promoting energy balance despite excessive caloric intake.

In humans, brown adipose tissue plays an important pyrogenic role at birth, but shrinks after birth, disintegrates to histologically rare, and becomes clinically insignificant in adults. However, recent findings overturn this idea and generate considerable interest in the role of brown adipose tissue in adulthood. In particular, by using a combination of positron emission tomography and automated X-ray tomography (PET-CT) to monitor tumor metastasis, it was accidentally discovered that a substantial proportion of highly active brown adipose depots in adults were estimated (Nedergaard). et al., 2007, Am J Physiol Endocrinol Metab 293:E444-E452). In a follow-up study, it was confirmed that in healthy adults these depots are in fact brown fats that function to express UCP-1 (Virtanen et al., 2009, N Engl J Med 360:1518-1525), in a cold environment but thermally. Brown-adipose-tissue activity was observed in more than 90% of young males exposed to neutral conditions (van Marken Lichtenbelt et al., 2009, N Engl J Med 360: 15000-1508). Moreover, in a retrospective analysis of nearly 2,000 PET-CT scans for various diagnostic reasons, the amount of active brown fat was inversely correlated with body mass index, a widely used measure of overall adiposity, suggesting an important role of brown fat in adult human metabolism. The likelihood is increased (Cypess et al., 2009, N Engl J Med 360:1509-1517). The role of pyrogenic adipocytes (eg, brown adipocytes or other UCP-1 expressing adipocytes) interspersed in white adipose tissue is somewhat less clear. Since pyrogenic adipocytes play an important role in metabolic activity, there is a need for a substance that increases exothermic adipocytes in vivo.

In addition, as obesity is emerging as a problem in modern society, many women in particular are making efforts to improve and shape their body. A well-balanced body and smooth skin can add not only attractiveness but also confidence to women. Now these needs extend from women to men. Body care includes not only obesity, but also obesity prevention and fat removal, as well as improving skin condition. In order to satisfy the needs of modern people, slimming body care products must be simple and effective, have no side effects on other organs, and ensure safety without affecting the human body even when used for a long period of time. Existing slimming body care products had the disadvantage of showing temporary effects such as the use of appetite suppressants or diuretics, and had side effects such as dizziness, vomiting, insomnia, and hallucinations. Also, conventionally known substances that promote methylxanthine-based fat breakdown, such as caffeine and theophylline, are recognized as addictive substances, and side effects such as bronchodilation and worsening of premenstrual syndrome have been reported, so there is no guarantee of safety. Therefore, it is urgently required to develop a new material that uses natural ingredients to exhibit the same or higher effect than conventional cosmetic products and is safer for the human body.

On the other hand, Schizandra chinensis Baillon. is a deciduous broad-leaved vine with alternate phyllotaxis, ovate or obovate leaves, sharply pointed tips, sawtooths, and some hairs on the veins on the back side. Flowers are red and white, bloom in June-July, and are bisexual. The fruit is used for food, hangs like an ear at the tip of the branch, and ripens red in August~September. The fruit is called Schisandra (Schizandrae fructus) and Schisandrae fructus. Jirisan, Jeollabuk-do, and Gangwon-do are the main production areas at an altitude of 200-1,600m, and they are wild all over the country except Chungnam and North Chungcheong. Geographically, they are distributed in Japan, Sakhalin, Manchuria, and China. In fruits, Schizandrol, Schizandrin A, B, C, angeloylgomisin O, P, epigomisin, pregomoisin, deoxyschizandrin ) and the like. Omija has long been used in herbal medicine as a raw material for herbal medicines because it has medicinal effects such as astringent, nourishing, tonic, anti-inflammatory, poisonous, thirsty, astringent gosap, ripe saengjin, and bosinyeomsim. The safety of the human body according to ingestion has already been confirmed.

Accordingly, the present inventors made efforts to find a natural product-derived anti-obesity agent. As a result, Schisandra extract increases the energy consumption of white adipocytes, induces the formation of brown adipocytes in white adipocytes, is involved in energy consumption, and is a marker gene for brown adipocytes. Since the expression of UCP-1 (Uncoupling protein-1), PGC-1α and PRDM16 was confirmed to be increased, the present invention was completed by confirming that the Schisandra extract can be used as an active ingredient in a composition for obesity.

The object of the present invention is Schisandra ( Schizandra ) chinensis ) to provide a pharmaceutical composition for preventing and treating obesity, a health functional food, a cosmetic composition, and an external preparation for skin containing the extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for inducing brown adipocytes, a health functional food, a cosmetic composition, and an external preparation for skin containing the Schisandra extract as an active ingredient.

In order to achieve the above object, the present invention is omija ( Schizandra chinensis ) extract as an active ingredient to provide a brown fat differentiation inducer.

In addition, the present invention provides a method for inducing brown adipocytes comprising the step of treating the extract of Schisandra Schisandra to white adipocytes.

In addition, the present invention provides a slimming (sliming) cosmetic composition containing Schisandra extract as an active ingredient.

In addition, the present invention provides a slimming skin external preparation containing Schisandra extract as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity containing the extract of Schisandra as an active ingredient.

In addition, the present invention provides a health functional food for preventing and improving obesity containing Schisandra extract as an active ingredient.

The present invention is Schisandra chinensis ) extract as an active ingredient, which relates to a pharmaceutical composition for preventing and treating obesity. Since it increases the expression of , the composition of the present invention can be used as a brown fat-inducing substance or anti-obesity substance, a pharmaceutical composition for inducing brown fat cells, a health functional food, a cosmetic composition, and a pharmaceutical composition for external application to the skin.

1 is a microscopic view showing the differentiation of preadipocytes extracted from inguinal white adipose tissue (iWAT) of mice and white adipocytes matured from preadipocytes.
Figure 2, Schisandra ( Schizandra ) chinensis ) is a diagram showing an increase in the expression of UCP-1 through the extract treatment.
3 is a diagram showing the increase in the expression of PGC-1α and PRDM16 through Schisandra extract treatment.

Hereinafter, the present invention will be described in detail.

The present invention is Schisandra chinensis ) extract as an active ingredient to provide a brown fat differentiation inducer.

The inducer preferably increases the expression of UCP-1 (Uncoupling protein-1), and preferably increases the expression of PGC-1α and PRDM16.

The Schisandra extract is preferably prepared by a manufacturing method comprising the following steps, but is not limited thereto:

1) extracting by adding an extraction solvent to Schisandra;

2) filtering the extract of step 1); and

3) Concentrating the filtered extract of step 2) under reduced pressure and drying it to prepare an extract of Omija.

In the above method, Schisandra of step 1) may be used without limitation, such as cultivated or commercially available ones. The omija fruit, seeds, leaves, stems or roots are all available, and most preferably a fruit, but is not limited thereto.

In the method, it is preferable to use water, alcohol, or a mixture thereof as the extraction solvent in step 1). It is preferable to use a C ₁ to C ₂ lower alcohol as the alcohol, and it is preferable to use ethanol or methanol as the lower alcohol. The extraction method may use an extraction method such as hot water extraction, immersion extraction, reflux cooling extraction, or ultrasonic extraction, but is not limited thereto.

It is preferable to extract the extraction solvent by adding 1 to 10 times to the amount of dried Schisandra, and more preferably adding 2 to 3 times to extract. The extraction temperature is preferably 20°C to 100°C, more preferably 20°C to 40°C, and most preferably room temperature, but is not limited thereto. In addition, the extraction time is preferably 10 to 48 hours, more preferably 15 to 30 hours, and most preferably 24 hours, but is not limited thereto. In addition, the extraction number is preferably 1 to 5 times, more preferably 3 to 4 times repeated extraction, and most preferably 3 times, but is not limited thereto.

In the above method, it is preferable to use a vacuum vacuum concentrator or a vacuum rotary evaporator for the vacuum concentration in step 3), but is not limited thereto. In addition, drying under reduced pressure, vacuum drying, boiling drying, spray drying or freeze drying is preferable, but is not limited thereto.

In a specific embodiment of the present invention, the present inventors treated the Schisandra extract to induce brown adipocytes in white adipocytes, which were extracted from inguinal white adipose tissue (iWAT), which is one of the subcutaneous fat tissues of mice. Preadipocytes were first cultured, and as a result, it was confirmed that white precursor cells were successfully extracted (see FIG. 1a ). To induce differentiation from the preadipocytes into adipocytes, the differentiation cocktail (IBMX 0.5 mM, Rosiglitazone 1 mM, Dexamethason 1 mM, insulin 5 mg/ml in 10% FBS DMEM/F12 GlutaMax media ) was treated for 2 days, and differentiation was carried out for a total of 6 days in a growth medium (10% FBS, insulin 5 mg/ml) every 2 days. As a result, it was confirmed that differentiation into mature white adipocytes was successfully induced (Fig. see 1b).

In addition, the present inventors have used it as a marker for inducing brown adipogenesis of white adipocytes, and observed an increase in UCP-1 expression, indicating that white adipocytes are becoming brown adipocytes and that energy consumption efficiency in adipocytes is increased. To measure, the expression level of the thermogenic gene (UCP-1) was analyzed using real-time PCR after 24 hours of treatment with a medium at a concentration of 1 μg/ml of Schisandra extract in white adipocytes. As a result, it was confirmed that the expression of UCP-1 in white adipocytes was increased more than two-fold due to the treatment of the Schisandra extract (see FIG. 2).

In addition, in order to measure the expression level of PGC-1α and PRDM16, which regulate the expression of UCP-1, which are thermogenic genes, the present inventors treated white adipocytes with a commercially available Schisandra extract and measured the expression level using real-time PCR. analyzed. As a result, it was confirmed that the Schisandra extract increased the expression of PGC-1α and PRDM16 (see FIG. 3 ).

Therefore, the Schisandra extract of the present invention increases the energy consumption of white adipocytes, induces the formation of brown adipocytes in white adipocytes, is involved in energy consumption, and contains uncoupling protein-1 (UCP-1), a marker gene for brown adipocytes; By confirming that the expression of PGC-1α and PRDM16 was increased, it was confirmed that the Schisandra extract can be used as a brown adipocyte differentiation inducer.

In addition, the present invention provides a method for inducing brown adipocytes comprising the step of treating the extract of Schisandra Schisandra to white adipocytes.

The composition preferably increases the expression of UCP-1 (Uncoupling protein-1), and preferably increases the expression of PGC-1α and PRDM16.

Schisandra extract of the present invention increases the energy consumption of white adipocytes, induces the formation of brown adipocytes in white adipocytes, is involved in energy consumption, and is a marker gene for brown adipocytes, UCP-1 (Uncoupling protein-1), PGC- By confirming that the expression of 1α and PRDM16 was increased, it was confirmed that the Schisandra extract can be used as a brown adipocyte differentiation inducer.

In addition, the present invention provides a slimming (sliming) cosmetic composition containing Schisandra extract as an active ingredient.

The composition preferably increases the expression of UCP-1 (Uncoupling protein-1), and preferably increases the expression of PGC-1α and PRDM16.

The composition includes skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, foundation, essence, nourishing essence, pack, soap, cleansing foam, cleansing lotion , it is preferable to have any one formulation selected from the group consisting of formulations of cleansing cream, body lotion and body cleanser, but is not limited thereto.

Schisandra extract of the present invention increases the energy consumption of white adipocytes, induces brown adipogenesis of white adipocytes, is involved in energy consumption, and UCP-1 (Uncoupling protein-1), PGC-, which are marker genes of brown adipocytes By confirming that the expression of 1α and PRDM16 was increased, it was confirmed that the Schisandra extract can be used as a slimming cosmetic composition.

The lipolytic agent contained in the slimming cosmetic composition according to the present invention may be commercially available domestically or internationally or prepared by a known method. According to the present invention, the slimming cosmetic composition comprising an extract extracted from Schisandra and a lipolytic agent as active ingredients has an excellent slimming effect on the abdomen or thigh.

Therefore, the slimming cosmetic composition containing the Schisandra extract and the lipolytic agent can be used as a composition for suppressing or treating obesity. The cosmetic composition may further include an active material constituting the composition as long as it does not have a detrimental effect on slimming, obesity suppression or treatment.

The formulation of the cosmetic composition is not particularly limited and may be appropriately selected according to the purpose. For example, it may be prepared in the form of a liquid, solid, cream, emulsion, paste, and the like. Specifically, the cosmetic composition of the present invention is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, It may be prepared in any one formulation selected from the group consisting of packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers.

When the formulation of the present invention is a paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

In addition, the slimming cosmetic composition of the present invention may contain other ingredients commonly formulated in cosmetics in addition to the ingredients described above as needed in each formulation. The ingredients that can be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, There are blood circulation promoters, cooling agents, limiting agents, thickeners, purified water, and the like.

In addition, the mixing|blending component which may be added is not limited to this, Any said component can be mix|blended within the range which does not impair the objective and effect of this invention. According to the present invention, the slimming cosmetic composition containing the extract obtained from Schisandra Schisandra using water and a lipolytic agent can be used as a food, pharmaceutical composition, etc. for the suppression or therapeutic effect of obesity.

In addition, the present invention provides an external preparation for slimming skin comprising the step of treating Schisandra extract to white fat cells.

The external preparation for skin preferably increases the expression of UCP-1 (Uncoupling protein-1), and preferably increases the expression of PGC-1α and PRDM16.

Schisandra extract of the present invention increases the energy consumption of white adipocytes, induces brown adipogenesis of white adipocytes, is involved in energy consumption, and UCP-1 (Uncoupling protein-1), PGC-, which are marker genes of brown adipocytes By confirming that the expression of 1α and PRDM16 was increased, it was confirmed that the Schisandra extract can be used as an external preparation for slimming skin.

In addition, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity containing the extract of Schisandra as an active ingredient.

The composition preferably increases the expression of UCP-1 (Uncoupling protein-1), and preferably increases the expression of PGC-1α and PRDM16.

Schisandra extract of the present invention increases the energy consumption of white adipocytes, induces brown adipogenesis of white adipocytes, is involved in energy consumption, and UCP-1 (Uncoupling protein-1), PGC-, which are marker genes of brown adipocytes By confirming that the expression of 1α and PRDM16 is increased, it was confirmed that the Schisandra extract can be used as a pharmaceutical composition for preventing and treating obesity.

The composition containing the extract of the present invention or an active fraction thereof may contain one or more active ingredients having the same or similar function in addition to the above ingredients.

The composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive includes starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, and lactose. , mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate , sucrose, dextrose, sorbitol and talc and the like can be used. The pharmaceutically acceptable additive according to the present invention is preferably included in an amount of 0.1 to 90 parts by weight based on the composition, but is not limited thereto.

That is, the composition of the present invention may be administered in various oral and parenteral formulations during actual clinical administration. In the case of formulation, commonly used fillers, extenders, binders, wetting agents, disintegrants, diluents or excipients such as surfactants It can be prepared using Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the mixed extract of the present invention, for example, starch, calcium carbonate, water It can be prepared by mixing cross (Sucrose), lactose (Lactose) or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Liquid formulations for oral use include suspensions, solutions, emulsions, and syrups, and various excipients such as wetting agents, sweetening agents, fragrances, and preservatives in addition to commonly used simple diluents such as water and liquid paraffin may be included. . Formulations for parenteral administration may 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.

The composition of the present invention may be administered orally or parenterally according to a desired method, and when administered parenterally, external skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection injection method It is preferable to select The dosage varies according to the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and severity of disease.

The dosage of the composition of the present invention varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of disease, and the daily dosage is the mixed extract of the present invention Based on the amount of 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, may be administered 1 to 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 for the prevention and treatment of asthma or inflammatory diseases.

In addition, the present invention provides a health functional food for preventing and improving obesity containing Schisandra extract as an active ingredient.

The composition preferably increases the expression of UCP-1 (Uncoupling protein-1), and preferably increases the expression of PGC-1α and PRDM16.

Schisandra extract of the present invention increases the energy consumption of white adipocytes, induces brown adipogenesis of white adipocytes, is involved in energy consumption, and UCP-1 (Uncoupling protein-1), PGC-, which are marker genes of brown adipocytes By confirming that the expression of 1α and PRDM16 is increased, it was confirmed that the Schisandra extract can be used as a health functional food for preventing and improving obesity.

The food composition containing the composition of the present invention as an active ingredient, each component contained in the extract of the present invention is a material that is safe for the human body, and even if it is ingested, it has an obesity suppression, therapeutic or slimming effect. It can also be used as a composition.

As food, for example, it can be used as various foods, drinks, gum, tea, vitamin complex, health functional foods, etc. The food can be prepared in dosage forms such as tablets, granules, powders, capsules, liquid solutions and pills according to a known manufacturing method, and the content of the extract according to the present invention is 0.0001 based on the total weight of the food composition depending on the dosage form. to 100% by weight. There is no particular limitation on other ingredients other than containing the extract according to the present invention as an active ingredient, and various conventional flavoring agents or natural carbohydrates may be included as additional ingredients.

Examples of the natural carbohydrate include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.

In addition, the food composition according to the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The proportion of these additives is not so critical, but may be selected from 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Therefore, the Schisandra extract of the present invention increases the energy consumption of white adipocytes and induces the brown adipogenesis of white adipocytes. By confirming that the expression of -1α and PRDM16 was increased, it was confirmed that the Schisandra extract can be used as an active ingredient in a brown adipocyte differentiation agent, a slimming cosmetic composition and external application for skin, and an anti-obesity composition and health functional food.

Hereinafter, the present invention will be described in detail by the following Examples and Formulation Examples.

However, the following Examples and Formulation Examples merely illustrate the present invention, and the content of the present invention is not limited by the following Examples and Formulation Examples.

< Example 1> Cell culture

According to the existing literature review, there is a report that brown localization occurs more in subcutaneous adipose tissue than in visceral fat among white adipose tissue. According to the present invention, pre-adipocytes extracted from inguinal white adipose tissue (iWAT), one of the subcutaneous fat tissues of mice, were first cultured as follows in order to induce brown adipocytes in white adipocytes by treatment with Schisandra extract. did

Specifically, the white adipose tissue near the groin of the rat was removed, washed in D-PBS, and then an isolation buffer (KRB buffer + 4% BSA + 1.5 mg/ml collagenase type 1) was added and sliced using scissors ( chopping). The Krebs-Ringer bicarbonate (KRB) buffer is 0.77 M NaCl 13.16 ml, 0.77 M KCl 0.52 ml, 0.77 M KH ₂ PO ₄ 0.132 ml, 0.77 M MgSO ₄ 0.132 ml, 0.77 M NaHCO ₃ 2.76 ml, 0.3 based on 100 ml M glucose 4.16 ml, 0.275 M CaCl ₂ 0.38 ml, DW 78.8 ml, adjusted to pH 7.4, and incubated in a 5% CO ₂ incubator for 20 to 30 minutes before use. The minced sample was left at 37° C. for 45 minutes, and then the sample was first filtered through a 300 μm nylon mesh and centrifuged at 200×g for 5 minutes. After removing the supernatant, the pellet was resuspended with D-PBS, passed through a 100 μm nylon mesh, and centrifuged again at 200×g for 5 minutes. After removal of the supernatant, cells resuspended in DMEM/F12 GultaMax media supplemented with 15% FBS were laid out in a 100 mm culture dish (Distinct transcriptional profiles of adipogenesis in vivo and in vitro. J Biol Chem. 2001, Soukas et al. )

As a result, as shown in FIG. 1A , it was confirmed that the white precursor cells were successfully extracted ( FIG. 1A ).

< Example 2> Confirmation of differentiation into white adipocytes

The following experiment was performed to induce differentiation of the preadipocytes prepared in <Example 1> into adipocytes.

Specifically, one day after the cells were 100% confluenced, they were treated with a differentiation cocktail (IBMX 0.5mM, Rosiglitazone 1 mM, Dexamethason 1 mM, insulin 5 mg/ml in 10% FBS DMEM/F12 GlutaMax media) for two days to grow at 2-day intervals. Differentiation was carried out in medium (10% FBS, insulin 5 mg/ml) for a total of 6 days (FGF21 regulates PGC-1α and browning of white adipose tissues in adaptive thermogenesis. Genes Dev. 2012, Fisher et al.).

As a result, it was confirmed that differentiation into successfully matured white adipocytes was induced as shown in FIG. 1B ( FIG. 1B ).

< Example 3> According to Schisandra extract treatment UCP -One( Uncoupling protein -1) Confirmation of increased protein expression

Uncoupling protein-1 (UCP-1) is known to increase energy consumption efficiency by increasing thermogenesis in the body. It has been reported that UCP-1 is rarely expressed in white adipocytes and is expressed in brown adipocytes responsible for energy consumption. Therefore, it is used as a marker for induction of brown adipocytes in white adipocytes, and the increase in UCP-1 in white adipocytes indicates that white adipocytes are becoming brown adipocytes and the energy consumption efficiency in adipocytes is increasing. The following experiment was performed to measure the expression level of Uncoupling protein-1 (UCP-1), a fever-related protein, due to the treatment of Schisandra extract in the differentiated white adipocytes prepared in <Example 2>.

Schisandra used in the present invention chinensis ( Turcz .) Baill . ) The extract is commercially available, and the detailed description of the product is shown in Table 1 below.

Specifically, in the differentiated white adipocytes prepared in <Example 2>, the level of expression of the thermogenic gene (UCP-1) 24 hours after treatment with the medium at a concentration of 1 μg/ml of the commercially available Schisandra extract was analyzed using real-time PCR. After separation from each sample using the RNeasy mini kit from Qiagen (USA), 2 μg of RNA and 2 μl of Oligo dT primer (500 μg/mL) were reacted at 70°C for 5 minutes, followed by RT of Bioneer (Korea) -CDNA was synthesized using a premix kit. Using the synthesized cDNA, 8 ng each was mixed with real-time PCR mix and reacted at 95°C for 15 minutes, followed by 40 cycles of 20 seconds at 95°C, 40 seconds at 60°C, and 15 seconds at 72°C. PCR was performed by reacting at 72°C for 5 minutes, at 60°C for 5 seconds, and at 95°C for 5 seconds. The primer sequences of the UCP-1 gene are shown in Table 2 below.

As a result, as shown in FIG. 2 , it was confirmed that the expression of UCP-1 in white adipocytes was increased more than two-fold due to the treatment of the Schisandra extract ( FIG. 2 ).

Schizandra berry PE 9% Schizandrins NATUREX Extracted from fruits together with seeds using alcohol and water as solvents. shape powder Color Brown incense Has a unique scent analysis Schisandra content > 9% by HPLC loss due to drying < 6% particle size 40 mesh 100% pass

< Example 4> Confirmation of increase in the expression of thermogenic genes following Schisandra extract treatment

There are various factors regulating the expression of UCP-1, a thermogenic gene, but PGC-1α and PRDM16 are representative. In the present invention, the following experiment was performed to measure the expression levels of PGC-1α and PRDM16, which are involved in the regulation of UCP-1 expression according to the treatment of Schisandra extract.

Specifically, as shown in <Example 3>, after treating the commercially available extract of Schisandra Schisandra to the differentiated white adipocytes prepared in <Example 2>, in order to examine the expression level of the thermogenic gene, Qiagen from each sample After separation using the RNeasy mini kit from (USA), 2 μg of RNA and 2 μl of Oligo dT primer (500 μg/mL) were reacted at 70°C for 5 minutes, and then using the RT-premix kit from Bioneer (Korea). to synthesize cDNA. Using the synthesized cDNA, 8 ng each was mixed with real-time PCR mix and reacted at 95°C for 15 minutes, followed by 40 cycles of 20 seconds at 95°C, 40 seconds at 60°C, and 15 seconds at 72°C. PCR was performed by reacting at 72°C for 5 minutes, at 60°C for 5 seconds, and at 95°C for 5 seconds. The primer sequences of each thermogenic gene are shown in Table 2 below.

As a result, as shown in FIG. 3 , it was confirmed that the Schisandra extract increased the expression of PGC-1α and PRDM16 ( FIG. 3 ).

Therefore, according to the present invention, it was found that the increase in the expression of UCP-1 in the Schisandra extract appears through the increase in the expression of the transcription factors of PGC-1α and PRDM16.

Primer name gene name base sequence SEQ ID NO: mCidea-F TGC TCT TCT GTA TCG CCC AGT SEQ ID NO: 1 mCidea-R GCC GTG TTA AGG AAT CTG CTG SEQ ID NO: 2 mDio2-F CAA CTG CCA GCC TTC CGC CA SEQ ID NO: 3 mDio2-R CAG GCA CTG CCC AGC CAT CC SEQ ID NO: 4 mUCP1-F UCP-1 CCC GCT GGA CAC TGC C SEQ ID NO: 5 mUCP1-R UCP-1 ACC TAA TGG TAC TGG AAG CCT GG SEQ ID NO: 6 prdm6-F PRDM16 CAG CAC GGT GAA GCC ATT C SEQ ID NO: 7 prdm6-R PRDM16 GCG TGC ATC CGC TTG TG SEQ ID NO: 8 mPGC1a-F PGC-1α CCC TGC CAT TGT TAA GAC C SEQ ID NO: 9 mPGC1a-R PGC-1α TGC TGC TGT TCC TGT TTT C SEQ ID NO: 10

A formulation example of the composition will be described below, but this is not intended to limit the present invention, but merely to describe it in detail.

< Formulation example 1> Cream making

Schisandra extract or fraction ....................10.00(%)

Polyethylene glycol monostearate ...............2.00

Self-emulsifying monostearate glycerin ...............................5.00

Cetyl Alcohol .................................................. .4.00

Squalene ............................................... ..6.00

Glyceryl tri2-ethylhexanoate ............................... 6.00

Sphingoglycolipids ............................................... 1.00

1.3-Butylene Glycol ............................................... 7.00

Vitamin C.................................................. ....1.00

The total amount was 100 with purified water, and a cream was prepared at the above compounding ratio (%).

< Formulation example 2> Preparation of tablets

Schisandra extract or fraction ....................50 mg

Cornstarch ............................................................... ..100 mg

Lactose ..................................................... .......100 mg

Magnesium Stearate ............................... 2 mg

Vitamin C.................................................. .....50 mg

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

< Formulation example 3> Manufacture of health drinks

Schisandra extract or fraction ....................1000 mg

Citric acid ............................................... ........1000 mg

oligosaccharide................................................. ........100 g

Plum Concentrate ............................................... .........2 g

Taurine ............................................................... ...............1 g

Purified water is added to the total ...............................900 ㎖

After mixing the above ingredients according to a conventional health drink manufacturing method, stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2L container, sealed and sterilized, then refrigerated and then stored in the present invention used in the manufacture of health beverage compositions.

Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demanding class, demanding country, and use.

Those of ordinary skill in the art to which the present invention pertains will be able to make various applications and modifications within the scope of the present invention based on the above contents.

<110> Korea Research Institute Bioscience and Biotechnology <120> Pharmaceutical composition for prevention or treatment of obesity comprising the extracts of Schizandra chinensis as active ingredient <130> 2014P-10-012 <160> 10 <170> KopatentIn 2.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> mCidea-F <400> 1 tgctcttctg tatcgcccag t 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> mCidea-R <400> 2 gccgtgttaa ggaatctgct g 21 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> mDio2-F <400> 3 caactgccag ccttccgcca 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> mDio2-R <400> 4 caggcactgc ccagccatcc 20 <210> 5 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> mUCP1-F <400> 5 cccgctggac actgcc 16 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> mUCP1-R <400> 6 acctaatggt actggaagcc tgg 23 <210> 7 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> prdm6-F <400> 7 cagcacggtg aagccattc 19 <210> 8 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> prdm6-R <400> 8 gcgtgcatcc gcttgtg 17 <210> 9 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> mPGC1a-F <400> 9 ccctgccatt gttaagacc 19 <210> 10 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> mPGC1a-R <400> 10 tgctgctgtt cctgttttc 19

Claims (11)

A cosmetic composition for inducing differentiation of brown fat, characterized in that it contains an extract of Schisandra chinensis and increases the expression of UCP-1, PGC-1α and PRDM16.
The cosmetic composition according to claim 1, wherein the extract is extracted with water, a C ₁ to C ₂ lower alcohol, or a mixed solvent thereof.
The cosmetic composition according to claim 1, wherein the UCP-1, PGC-1α and PRDM16 are marker genes for brown adipocytes.
The cosmetic composition according to claim 1, wherein the UCP-1 is a thermogenic gene, and the PGC-1α and PRDM16 are transcription factors that regulate UCP-1 expression.
According to claim 1, wherein the composition is skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, foundation, essence, nourishing essence, pack, soap A cosmetic composition, characterized in that it has any one formulation selected from the group consisting of formulations of , cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser. delete delete delete delete delete delete

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