KR101650377B1 - Compositions for Preventing or Treating Obesity Comprising Kahweol - Google Patents

Abstract

The present invention relates to a composition for preventing or treating obesity, which comprises Kahweol, wherein the composition for preventing or treating obesity comprising an effective ingredient of kahweol of the present invention has no side effects caused by conventional synthetic pharmaceutical compositions , Lipid precursor cells to differentiate into adipocytes and inhibit proliferation of adipocytes as well as lipid accumulation in adipocytes. Therefore, it is useful as a pharmaceutical composition for preventing or treating obesity, or as a health functional food for preventing or improving obesity Can be utilized.

Description

[0001] Compositions for Preventing or Treating Obesity [

The present invention relates to a composition for preventing or treating obesity, which comprises Kahweol, and more particularly to a composition for prevention or treatment of obesity comprising Kahwol derived from coffee having an effect of inhibiting differentiation into adipocytes and inhibiting lipid accumulation in adipocytes To a pharmaceutical composition for preventing or treating obesity or a health functional food.

Along with the recent westernized diet, obesity, which is a state of excessive accumulation of fat tissue, is increasing due to the imbalance of calorie intake and consumption. Obesity is an independent or closely related to other diseases leading to serious health problems, in particular increasing the prevalence and mortality of colorectal, ovarian, breast, prostate, and other cancers, including diabetes, diabetes, degenerative arthritis, Impoverishment and psychological disorders. Therefore, the prevention and treatment of obesity is considered to be a very effective means for prevention of cancer and various adult diseases, and attention is gathering worldwide.

Obesity is a well-known risk factor that causes many ailments, including atherosclerosis, hypertension, diabetes, stroke, pulmonary embolism, sleep apnea and cancer. In addition, it complicates a number of clinical diseases such as respiratory diseases, osteoarthritis, osteoporosis, gallbladder disease and dyslipidemia. The severity of this problem is best reflected in the fact that the mortality rate increases with weight gain. As with 35 million Americans, if the BMI exceeds 30 kg / m 2, all causes of death with a mortality rate above 50% can be attributed to obesity-related illnesses.

Obesity is mostly consumed in the calories consumed and the remaining part is converted into adipocyte and accumulated in the subcutaneous tissue and abdominal cavity. Since there is a limit to the size of the adipocytes, it is necessary to increase the number of adipocytes in order to store the energy quickly when consuming the excess energy, and thus, a new adipocyte is produced in the human body. The new adipocytes differentiate from pro-adipocytes, which are formed by differentiation from stem cells. The process of differentiation of stem cells into adipocytes through adipocytes is called hormone, growth Factors such as urea, cytokine, and the like regulators are affected.

Commercial anti-obesity agents include thiazolidinediones (TZDs), Xenical (Korean Roche) and sibutramine. The thiazolidinediones (TZDs) inhibit the formation of sugars, and Xenical (Korean rosacea) and sibutramine inhibit the absorption of fat and suppress the reabsorption of serotonin in synapses, .

However, these substances were originally used as antidepressants and caused serious side effects. In addition to these, anti-obesity agent called adipex is an appetite suppressant that contains pentamamine with sympathetic stimulant activity, and has a side effect by controlling the neurotransmitters associated with the hypothalamus.

Side effects of these drugs have been reported to include cardiovascular, central nervous, hepatic and renal disorders. Therefore, there is an urgent need to develop high value-added, multi-functional products that prevent side effects from long-term ingestion and prevent and treat obesity. Various herbal medicine extracts have been disclosed which minimize various side effects such as cardiovascular, central nervous, hepatic and renal disorders.

However, these herbal medicine extracts have a limited effect on reducing fat cells and inhibiting the production of triglycerides and thus being used as a composition for preventing and treating obesity and lipid-related metabolic diseases.

On the other hand, kahweol is known as a substance present in coffee, and coffee has potentials such as two diterpenes including chlorogenic acid, caffeine and cafestol and kahweol. Thousands of compounds containing bioactives are included. A typical Arabica coffee bean includes a cafe stall and its structural analogue caywal, and kahweol is a fat soluble compound known as a diterpenes molecule and is present in an oil derived from coffee bean.

In the prior art, Caywal has been shown to be effective against cancer (Korean Patent Publication No. 2006-0001162), anti-inflammatory (Korea Patent Publication No. 2005-0107876), angiogenesis inhibition (Korean Patent Publication No. 2006-0001161) Korean Patent Laid-Open Publication No. 2013-0119032), etc., and coffee extracts or other compounds are known to be effective for inhibiting obesity (Korean Patent Publication No. 2004-0036112; 2011-0087969), there has been no research that can be used as a preventive and therapeutic agent for obesity.

Therefore, in the present invention, as a result of intensive efforts to select compounds affecting the differentiation into adipocytes among the compounds derived from various natural extracts, it has been found that, in addition to the inhibition of the differentiation of coffee-derived camellia into adipocytes, And thus can be used as a composition for preventing or treating obesity. Thus, the present invention has been completed.

Disclosure of the Invention The present invention has been conceived to solve the above problems, and it is an object of the present invention to provide a pharmaceutical composition for preventing or treating obesity comprising an effective ingredient of kahweol or a health functional food for preventing or improving obesity.

In order to solve the problems of the present invention, the present invention includes kahweol as a pharmaceutical composition for preventing or treating obesity containing an active ingredient, or a health functional food for preventing or improving obesity.

According to a preferred embodiment of the present invention, the cowweed is a coffee-derived compound, and may include a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

According to another preferred embodiment of the present invention, the cowl may be included in an amount of 0.001 to 10 parts by weight based on 100 parts by weight of the total composition.

According to another preferred embodiment of the present invention, the canola may inhibit adipocyte differentiation and accumulation of fat in adipocytes.

The composition for prevention or treatment of obesity comprising kahweol of the present invention as an active ingredient has no adverse effects caused by the conventional synthetic pharmaceutical composition and can be used not only for differentiation into adipocytes and inhibiting proliferation of adipocytes, It can be effectively utilized as a pharmaceutical composition for preventing or treating obesity or as a health functional food for preventing or improving obesity.

Figure 1 shows the results of library screening of natural products affecting differentiation into adipocytes.
Fig. 2 shows data on the degree of differentiation of adipocytes (A) and the degree of lipid accumulation in adipocytes (B) according to the addition of cowork.
FIG. 3 shows data on the degree of differentiation of adipocytes (A) and the degree of lipid accumulation in adipocytes (B) according to the addition of cafestol.

Hereinafter, the present invention will be described in more detail.

As described above, drugs used as preventive or therapeutic agents for obesity cause heart diseases, respiratory diseases, elevated blood pressure and insomnia as side effects of drugs and low persistency of their efficacy. Therefore, there is an urgent need to develop high value-added, multi-functional products which have no side effects, prevent and treat obesity, reduce oxidative stress, and prevent diverse diseases.

The present invention sought to solve the above-mentioned problems by providing a composition for preventing or treating obesity comprising an effective ingredient of kahweol. The present invention provides a method for preventing obesity, which is capable of specifically reducing adipocytes, inhibiting the differentiation of adipocytes into adipocytes, and reducing the accumulation of fat in adipocytes, Or a therapeutic pharmaceutical composition or a health functional food for preventing or ameliorating obesity.

Accordingly, the present invention includes kahweol as a pharmaceutical composition for preventing or treating obesity comprising an active ingredient, or a health functional food for preventing or improving obesity.

The cowweed is a diterpenes-based compound, which may include a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof, and may be characterized in that it is derived from coffee, preferably coffee bean have.

[Chemical Formula 1]

The canola may be isolated from coffee beans by methods known in the art and may be obtained from refining in a unsaponifiable material, especially in the oil of coffee beans (coffee bean), but is typically prepared and / And is not particularly limited as long as it can.

In one embodiment of the present invention, among the compounds derived from various natural extracts, compounds that affect the differentiation into adipocytes were selected, and as a result, phorbol 12-myristate 13-acetate (PMA) , 4α-phorbol 12-myristate 13-acetate, piceatannol, prostaglandin A1, kahweol acetate, rapamycin, taxol, Vincristine sulfate and vinpocetin were firstly screened, and finally, cellulosic and non-cytotoxic carnis were selected.

In the present invention, the cowweed may be contained in an amount of 0.001 to 10 parts by weight, preferably 0.001 to 5 parts by weight, more preferably 0.002 to 1 part by weight, based on 100 parts by weight of the total composition, Ml to 10 mg / ml. In addition, the canola can inhibit adipocyte differentiation, proliferation inhibition and fat accumulation in adipocytes.

The above-mentioned carnitine can be taken in an appropriate amount when it is applied to food or the like, and the proper amount can be set in consideration of the concentration of the carnol established in in vitro conditions such as cell experiments.

 The solvent of the composition is not particularly limited as long as it does not interfere with the effect of inhibiting the differentiation of adipocytes in water, more preferably water, ethanol, propyl alcohol, DMSO, and the like. In addition, when the carnitine is contained at a concentration lower than 0.001 part by weight with respect to 100 parts by weight of the total composition, the adipocyte differentiation backbiting efficacy may be decreased, and when the concentration is higher than 10 parts by weight, It may not appear to be superior.

In one embodiment of the present invention, the inhibitory effect on adipocyte differentiation and the inhibitory effect on the lipid accumulation in adipocytes according to the treatment concentration of the carnitine showed that inhibition of adipocyte differentiation was dependent on the concentration of carnitine as shown in Fig. 2 It was confirmed that the treatment of 25 / / ml significantly inhibited the differentiation of adipocytes (FIG. 2A). In addition, lipid accumulated in adipocytes was also found to be inhibited in a concentration dependent manner on the basis of cagwort concentration. When treated with 25 ㎍ / ㎖, lipid accumulation was reduced by about 40 to 50% Respectively.

In another embodiment of the present invention, inhibition of differentiation into lipoprotein and inhibition of lipid accumulation was confirmed by using cefestol, a diterpenes compound having a structure similar to that of camellia, As described above, it was confirmed that cafe stall had no inhibitory effect on lipid accumulation and inhibition of differentiation into adipocytes.

In other words, most of the diterpenes compounds belonging to the cayworms can not be considered to inhibit adipocyte differentiation and inhibit lipid accumulation, and inhibition of adipocyte differentiation and inhibition of lipid accumulation can be regarded as a unique function of the cayworm.

Therefore, the composition for inhibiting adipocyte differentiation and proliferation comprising the cowyl of the present invention as an active ingredient can be used not only as side effects of the conventional pharmaceutical composition, but also as an agent for inhibiting proliferation of adipocyte differentiation and adipocyte proliferation, The lipid accumulation in the adipocyte can be effectively inhibited, so that the carnitine can be effectively utilized as a pharmaceutical composition for preventing or treating obesity, or as a health functional food for preventing or improving obesity.

The pharmaceutical composition for preventing or treating obesity, which comprises the cayworm according to the present invention as an active ingredient, may further comprise other natural substances or compounds having an anti-obesity effect. The pharmaceutical composition of the present invention may be used in rats, mice, , Human, and the like, via various routes including oral, transdermal, subcutaneous, intravenous, or muscular. In addition, the pharmaceutical composition for preventing or treating obesity according to the present invention can be formulated into various formulations. In the case of formulation, it may be formulated using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants which are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which are mixed with at least one excipient (e.g., starch, sucrose, lactose and gelatin) Can be prepared. In addition to simple excipients, lubricants can also be used. Examples of liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances and preservatives . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The pharmaceutical composition for prevention or treatment of obesity including the cowwheel according to the present invention may vary depending on the age, sex and body weight of the patient, but is generally in an amount of 0.01 to 50 mg / kg, preferably 0.1 to 10 mg / kg May be administered once or several times a day, and the concentration may be set in consideration of the concentration of the carnitine established in an in vitro experiment such as a cell experiment.

Also, the dosage of the composition containing the carnitine may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age and the like. Accordingly, the dosage is not limited in any way to the scope of the present invention.

In addition, the kind of the health functional food including the health functional food composition for prevention or improvement of obesity including the cowl of the present invention is not particularly limited, and examples thereof include meat, sausage, bread, chocolate, candy, snack, , Noodles, other noodles, gums, dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes.

 The health food may be used in combination with other food or food additives other than the above-mentioned cabal, and may be suitably used according to a conventional method. For example, a non-alcoholic fatty liver containing beverage containing the above-mentioned cayworm as an active ingredient may be prepared by adding a calcium, an ascorbic acid concentrate, a liquid fructose, purified water and the like in addition to the cayworm as an active ingredient, And then cooled to room temperature to prepare a beverage. In addition, the non-alcoholic health supplement for preventing obesity which contains the above-mentioned carnol as an active ingredient is prepared by adding nutritional supplement components (vitamin B1, B2, B5, B6, E and acetic acid ester, nicotinic amide), oligosaccharide, 50% ethanol, After mixing in a granular form, it is dried in a vacuum drier and then passed through 12 to 14 mesh to uniformly prepare granules. The granules are then extruded in suitable amounts to be refined or powdered or filled into hard capsules. Can be manufactured.

The effective dose of the cowweed contained in the health food can be used in accordance with the effective dose of the pharmaceutical composition, and the amount of the active ingredient to be mixed can be suitably determined according to the purpose of use such as preventive or therapeutic treatment. In the case of long-term consumption intended for health and hygiene purposes or for health control purposes, it may be below the above range.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Selection of compounds derived from natural extracts with inhibitory effect on adipocyte differentiation

In the present invention, screening was carried out to select a substance capable of specifically inhibiting adipocyte differentiation among the various natural-substance-derived compounds shown in FIG. 1, and each compound can be purchased on the market. Enzo Life Science Science, USA).

First, the preadipocytes (3T3-L1, Korea Cell Line Bank, cat. 10092.1) a 5% CO _2, 37 ℃ seeded into 8 × 10 ³ cell / well in 48-well plates (48-well plate) for 48 hours. And incubated in an incubator. After 48 hours, when the cell density reaches 100%, the culture solution is removed, and a new culture solution is dispensed and cultured for another 48 hours. After the culture, the culture medium was removed and differentiation was induced for 3 days using an adipocyte differentiation medium (DMEM containing 0.5 mM IBMX (3-Isobutyl-1-methylxanthine) and 1 μM Dexamenthason and 5 μg / ml Insulin). After 3 days, the culture medium was removed, and the various natural product-derived compounds shown in Fig. 1 were treated at a concentration of 25 占 퐂 / ml in a medium containing 5 占 퐂 / ml of insulin. As a control. Thereafter, the medium containing 25 μg / ml of insulin and 5 μg / ml of the compound derived from natural products was continuously distributed every 2 days for 8 days

The cells were then washed with phosphate buffered saline (PBS), and then 200 μl of 10% formaldehyde was added to the wells. The cells were fixed at room temperature for 1 hour, and formaldehyde , Washed with tertiary distilled water, washed with 60% isopropanol and dried. After the plate was dried, 200 ㎕ of Oil red O dye was added, and the plate was stained at room temperature for 1 hour and then washed three times with distilled water. The stained cells were observed with a microscope and the degree of differentiation from adipocytes into adipocytes was observed.

The oil red ophthalmic solution is a solution obtained by dissolving 350 mg of Oil Red O dye in 100 ml of isopropanol in a ratio of 6: 4 with distilled water, and then filtering with a 0.22 탆 filter.

As a result, as shown in Fig. 1, phorbol 12-myristate 13-acetate (PMA), 4α-phorbol 12-myristate 13-acetate (PMA) piceatannol, prostaglandin A1, kahweol acetate, rapamycin, taxol, vincristine sulfate and vinpocetin were first selected, and finally, Cytolytic cytokines were selected for the differentiation of adipocytes.

Inhibition of adipocyte differentiation by cowworm and inhibition of lipid accumulation in adipocytes

In order to confirm the effect of kahweol (cat. ALX-350-223-M010, Enzo Life Sciences) on adipocyte differentiation and adipocyte accumulation, adipose precursor cells cultured in the same manner as in Example 1 (0, 12.5 and 25 占 퐂 / ml) to induce differentiation into adipocytes.

The induction and observation of adipocyte were carried out in the same manner as in Example 1. After observing with a microscope to confirm the degree of lipid accumulation in adipocytes, 300 μl of isopropanol per well was used to extract the staining dye in the adipocyte, The optical density value (OD value) was measured at 500 nm.

As a result, as shown in FIG. 2, it was confirmed that the differentiation into adipocytes was inhibited depending on the concentration of the carnitine. Particularly, when 25 μg / ml was treated, the adipocyte differentiation was remarkably inhibited (FIG. 2A). In addition, it was confirmed that the fat accumulated in the adipocyte was also inhibited in a concentration dependent manner on the cowwort. When 25 μg / ml was treated, the fat accumulation was reduced by about 40 to 50% as compared with the control without cowwort treatment Respectively.

Diterpenes  Compound phosphorus In the cafestol (cafestol)  To inhibit adipocyte differentiation and to inhibit lipid accumulation in adipocytes

ALP-350-220-M050, Enzo Life Sciences), which is a diterpene compound, has similar inhibitory effects on lipid peroxidation and lipid accumulation inhibition , ≪ / RTI >

As a result, as shown in Fig. 3, it was confirmed that caffeolol had no inhibitory effect on the differentiation into adipocytes and lipid accumulation inhibitory effect.

That is, the diterpenes compound belonging to the cowworm can not be considered to inhibit adipocyte differentiation and inhibit lipid accumulation, and it is confirmed that the inhibition of adipocyte differentiation and the inhibition of lipid accumulation are unique functions of the carnitine.

Claims (8)

A composition for inhibiting adipocyte differentiation in vitro comprising kahweol at an effective concentration of 22 占 퐂 / ml to 28 占 퐂 / ml.
The method of claim 1, wherein the cowweed is a coffee-derived compound,
A composition for inhibiting adipocyte differentiation in vitro, which comprises a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.
[Chemical Formula 1]

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