KR20140124480A - Composion comprising extract of Angelica tenuissima Nakai for controlling ipid metabolism - Google Patents

Abstract

The present invention relates to extract of Angelica tenuissima Nakai having the effect of improving lipid metabolism and a composition containing the same for controlling lipid metabolism and, more specifically, to a composition for controlling lipid metabolism, which is highly effective in adjust or improving lipid metabolism by containing extract of Angelica tenuissima Nakai which has pancreatic lipase-inhibiting activity and can reduce a lipid amount and a neutral lipid amount by suppressing adipocytic differentiation and lipid metabolism.

Description

[0001] The present invention relates to a composition for controlling lipid metabolism,

The present invention relates to a hyperglycemic extract having lipid metabolism improving effect and a composition for controlling lipid metabolism containing the same, which has a pancreatic lipase inhibitory activity and contains a hypertonic extract capable of reducing lipid production and neutral mass in 3T3-L1 adipocytes To a composition for controlling lipid metabolism, which has an excellent effect for controlling or improving lipid metabolism.

In modern society, diverse metabolic diseases such as obesity, diabetes, hypertension and arteriosclerosis are rapidly increasing due to repeated lifestyle such as high calorie diet, lack of exercise, excessive stress such as high fat, high sugar and high salt. In particular, obesity is an energy metabolism abnormality caused by an imbalance between intake energy and consumed energy, which is defined as a state in which excess fat is accumulated in fat cells. The most common cause of obesity is increased body weight or body fat accumulation due to ingestion of food containing high energy or high fat, lack of exercise, or body fat accumulation. Cytologically, obesity is caused by the differentiation of adipocyte and lipid metabolism, Of the world.

It has been reported that obesity increases the risk of severe adult diseases such as hypertension, type 2 diabetes, hypertension, heart disease, stroke, arthritis, arteriosclerosis and cancer, as well as external problems. Recently, studies on the prevention and treatment of obesity have been actively carried out. These studies mainly focus on inhibition of differentiation of adipocytes, suppression of lipid accumulation in adipocytes, and decomposition of accumulated lipids.

Meanwhile, as a principle of treatment for obesity, diet and exercise are the most appropriate methods. Recently, development of appetite suppressant, heat production promoter and absorption inhibitor have been progressed, and the usefulness of anti-hyperlipidemia has been attracting attention. Among them, lipase inhibition Pancreatic lipase inhibitors have been attracting interest as a way to slow the absorption of fatty acids. Pancreatic lipase acts as a key enzyme that degrades triglyceride into 2-monoacylglycerol and fatty acid. Therefore, if there is a substance that specifically inhibits pancreatic lipase, it may be a useful therapeutic agent for obesity. A representative example of such a pancreatic lipase inhibitor is tetrahydrolipstatin (orlistat), a derivative of lipstatin derived from Streptomyces toxitricini, which is known to have the highest efficacy to inhibit about 30% of the fat ingested and is currently marketed as a pharmaceutical product . However, despite these benefits, orlistat has been known to have side effects such as gastrointestinal disorders, hypersensitivity, biliary secretion, and lipid-soluble vitamin absorption inhibition. Recently, studies for the development of pancreatic lipase inhibitors from natural materials without side effects have been conducted.

Such techniques include Korean Patent Publication No. 2003-0088528 (a method for inhibiting pancreatic lipase using eggplant extracts and a composition for inhibiting the absorption of fat by the body using the same), Korean Patent Publication No. 2006-0120791 (lipid Metabolic improvement and pharmaceutical composition for prevention and treatment of obesity). However, these natural extracts were limited in their ability to improve lipid metabolism.

It is an object of the present invention to provide an extract derived from a natural product, which has excellent lipid metabolism control or improving effect.

In addition, the present invention provides a ganoderma extract which is excellent in pancreatic lipase inhibitory activity and can effectively reduce lipid production and neutral mass in 3T3-L1 adipocytes, and can be used as a food or pharmacological agent And to provide a composition.

In order to achieve the above object, the present invention provides a food composition for improving lipid metabolism, which comprises an extract of Angelica tenuissima Nakai.

The present invention also provides a pharmaceutical composition for preventing or treating lipid metabolism-related diseases, which comprises an extract of Angelica tenuissima Nakai.

Hereinafter, means for solving the problems of the present invention will be described in detail.

The present invention relates to a food composition for improving lipid metabolism or a pharmaceutical composition for the prevention or treatment of lipid metabolism-related diseases, which comprises pancreatic lipase inhibitory activity by containing a goat extract as an active ingredient, And the mass of the neutral region, and can be effectively used for controlling or improving lipid metabolism.

The vertebrate ( Angelica tenuissima Nakai) used in the present invention means a perennial herbaceous herbaceous perennial plant of the dicotyledonous plant, and its photograph is shown in Fig. It is used for headache, fever, seawater, sputum, runny nose, etc., and it is used for limb paralysis joint pain. Especially, it is known that it has efficacy in headache of crown palsy. Has calming, analgesic, antipyretic, anti-inflammatory action, inhibits the intestinal and uterine smooth muscle, and is known to have a strong inhibitory action against Pycnogenetics.

The extract of the present invention can be obtained through various methods known in the art. Examples of the extraction solvent include purified water, methanol, ethanol, butanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, And mixtures thereof.

In addition, the gum extract of the present invention includes not only the above-mentioned extraction method, but also extracts extracted by other methods. For example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value; Separation by chromatography made for separation by various chromatographies, for example size, charge, hydrophobicity or affinity; The fractions obtained through various purification methods which are additionally included are also included in the extract of the present invention.

In addition, the gum extract of the present invention may be used in a liquid state, but may be prepared in powder form by an additional process such as vacuum distillation, freeze-drying, or spray drying.

As shown in the following examples, the extract of Ganoderma extract of the present invention has pancreatic lipase inhibitory activity and can be effectively used for controlling or improving lipid metabolism by decreasing lipid production amount and neutral mass in 3T3-L1 adipocytes .

On the other hand, the food composition containing the guinea pig extract of the present invention can be used as a health functional food in powder, granule, tablet, capsule or beverage form. At this time, the food composition of the present invention may contain other ingredients which can give a synergistic effect to the main effect, within the range not impairing the intended main effect of the present invention, in addition to the above- Various food-acceptable food additives may be added. Preferably, depending on the form of the desired composition, it may be added with conventional additives or additives such as sweeteners such as licorice, vitamin C, citric acid, nicotinic acid, sodium benzoate, aspartame, saccharin, pectin, malitol, sorbitol, xylitol, guar gum, One or more components selected from the group consisting of milk powder and oligosaccharides may be added to increase taste and aesthetics.

The food composition of the present invention may be added to various foods for the development of health supplement foods such as various foods, meat, beverages, chocolate, snacks, confectionery, pizza, ramen, other noodles, Alcoholic beverages, vitamin complexes, health supplements and the like.

Meanwhile, the pharmaceutical composition for preventing or treating lipid metabolism-related diseases containing the extract of Ganoderma extract of the present invention is useful for prevention of lipid metabolism-related diseases such as obesity, diabetes, hypercholesterolemia, hyperlipidemia, Or for therapeutic use. In this case, the pharmaceutical composition of the present invention can be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or other oral formulations, external preparation, have.

In addition, the pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the production of pharmaceutical compositions. Such carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The dosage of the pharmaceutical composition of the present invention is preferably determined in consideration of the administration method, the age, sex, and weight of the recipient, severity of the disease, and the like. For example, the pharmaceutical composition of the present invention can be administered at least once at a dose of 0.1 to 100 mg / kg (body weight) per day based on the active ingredient. However, the dosage is only an example and may be changed by a doctor's prescription depending on the condition of the recipient.

According to the present invention, it is possible to provide a food or pharmaceutical composition having excellent pancreatic lipase inhibiting activity and having an excellent effect for improving lipid metabolism by using a ganoderma extract which can effectively reduce lipid production and neutral mass in 3T3-L1 adipocytes have.

Fig. 1 is a photograph of a copy of the present invention.
Fig. 2 is a process diagram showing the fractionation process of the ginseng extract (hot water extract) of the present invention.
3 is an HPLC chromatogram of a blank.
4 is an HPLC chromatogram of the hot water extract of the present invention.
Figure 5 is an HPLC chromatogram of the goblet ethyl acetate fraction of the present invention.
Figure 6 is an HPLC chromatogram of the butanol fraction of the present invention.
Figure 7 is an HPLC chromatogram of the purified water fraction of the present invention.
FIG. 8 is a graph showing the effect of inhibiting lipid production upon addition of the extract of hot pepper (hot water extract) of the present invention.
9 is a graph showing the change in the mass of the neutral paper according to the addition of the extract of hot pepper (hot water extract) of the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

Example 1: Preparation of goat extract and general component analysis of goat

200 g of Gobbon was dissolved in 1,000 mL of distilled water and extracted with 95 ° C hot water for 6 hours . The extraction yield was 45.13 ± 3.09%.

The general analysis of Ganoderma lucidum extract was carried out according to the AOAC method. That is, the moisture content was 105 ° C, the dry heat drying method, and the ash content method was a dry conversation method in which the sample was measured at 550 ° C. Soxhlet method was used for crude fat content and Kjeldahl method was used for crude protein content. Each sample was repeated three times and expressed as a mean value. The results are shown in Table 1 below.

According to Table 1, 9.49% of moisture, 5.13% of crude protein, 10.51% of crude fat and 15.06% of crude protein were detected in the sample. From these results, it was found that the contents of crude protein, crude protein and crude fat were high.

Example 2: Preparation of gumbo fraction

For the solvent fraction of the goblet, an organic solvent fraction of polarity was carried out with 95 캜 hot water extract. As the organic solvent used in the fractionation, three kinds of fractions were obtained including the organic solvent fraction and the water fraction of polar ethyl acetate and butanol.

In other words, 200 g of goads were dissolved in 1,000 mL of distilled water, and the resulting extract was subjected to hot water extraction at 95 ° C for 3 hours and repeated for 6 hours. The extract was concentrated using a rotary vacuum evaporator (Tokyo rikakikai CO., Japan), and the extract was applied to a fractionation funnel After re-dissolving in distilled water, 500 mL of polar organic solvent was added, and the sequential fraction was repeated three times and concentrated under reduced pressure. The yield of each fraction is shown in Fig.

Example 3: Screening of analytical conditions of goat extract and fractions

HPLC (LC-20A, Shimadzu Corp., Japan) was used to investigate the analytical conditions of the gibberellins. The extracts and fractions were diluted to a concentration of 10 mg / mL and analyzed for retention time.

The analytical column was a Luna C ₁₈ column (Phenomenex, USA, 10 μm, 250 × 4.6 mm). The mobile phase A used was water and the mobile phase B was acetonitrile. The gradient conditions of the mobile solvent for each sample analysis are shown in Table 2 below, and the compounds coming out through the column for 41 minutes were monitored at 254 nm and analyzed. The elution rate of the mobile solvent was maintained at 1.0 mL / min and the temperature of the oven was maintained at 40 ° C. The sample was filtered with a 0.45 μm syringe filter (Advantec, Japan) and analyzed by injection into HPLC.

The results are shown in Figs. Here, blank was analyzed using the third distilled water under the same conditions. Figs. 3 to 7 are chromatograms of blank, hot water extracts of 95 占 폚, ethyl acetate fraction of goat, butanol fraction of goat, and water fraction of goat. The ethyl acetate fraction showed the strongest peak between 17 and 18 minutes, the butanol fraction showed the strongest peak between 8 and 10 minutes, and the same peak appeared in the ethyl acetate fraction.

Example 4: Pancreatic lipase inhibitory activity of goatee extract and fractions

In this Example, pancreatic lipase inhibitory activity of goat extract and fractions was measured.

The porcine pancreatic lipase was dissolved in 0.5 g / 200 mL of the enzyme buffer (10 mM MOPS, 1 mM EDTA, pH 6.8) at 4 ° C and centrifuged at 4000 rpm. The supernatant was transferred to 169 μl of Tris buffer (100 mM Tris-HCl, 5 mM CaCla 2, pH 7.0) and 6 μl of enzyme buffer. Samples were dissolved in DMSO to a final concentration of 3% and diluted to various concentrations. After dissolving p- nitriphenyl butyrate ( p -NPB) as a substrate solution in DMF for 10 mM, the enzyme and sample were shaking incubated at 37 ° C for 15 min. Substrate was added and shaking incubated at 37 ° C for 30 min. Absorbance was measured at 405 nm using an ELISA raider.

The inhibitory activity of pancreatic lipase was shown by the absorbance reduction rate of the group of addition of the sample solution and the group of no addition, and the results are shown in Table 3 below.

According to the above Table 3, it was found that the pancreatic lipase inhibitory activity of the whole lotus appeared in the order of ethyl acetate fraction, butanol fraction, hot water extract and water fraction.

Example 5: Ability to inhibit fat accumulation

In this example, the ability to inhibit fat accumulation in the goat was measured.

Cultures of 3T3-L1 adipose precursor cells were cultured in DMEM supplemented with 1% penicillin-streptomycin (P / S) and 10% bovine calf serum (BCS) at 37 ° C and 5% CO ₂ . Cells were washed with phosphate buffered saline (PBS) at 70% on the bottom of 100 plates, and incubated for 3 min at 37 ° C and 5% CO _{2 with} 0.25% trypsin-EDTA solution. Cells were desorbed and subcultured for maintenance. To induce the differentiation of 3T3-L1 adipose precursor cells, the cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% P / S at 1.25 × 10 ⁵ cells / well in each well of a 6-well plate After 2 days, the medium was exchanged, and the cells were completely fused on the 3rd to 4th day. Differentiation was induced by treatment with 10% FBS and MDI solution (0.5 mM 3-iso-Butyl-1-methylxanthine, 1 M dexamethsone, 5 / mL insulin) in the fusion state.

In order to observe the effect of adipocyte differentiation on adipocyte differentiation, ganoderma extract (hot water extract) was treated at different concentrations. After 2 days of induction of differentiation, medium was replaced with DMEM containing 10% FBS, 1% P / S and 5 / mL insulin, and 10% FBS and 1% P / S Cells were differentiated by exchanging with included DMEM.

Oil Red O Staining was carried out using cells that were 7 to 9 days after induction of differentiation. After removing 10% FBS DMEM, the cells were washed twice with PBS, fixed with 10% formalin at 4 ° C for 1 hour, and then washed three times with distilled water. The washed cells were stained with Oil Red O, which specifically reacts with the intracellular lipid for 1 hour. After staining, the staining solution was removed, washed three times with distilled water, and then distilled water was filled in wells and observed under a microscope.

For determination of triglyceride, distilled water was removed and 2 mL of 100% iso-propyl alcohol was added to the completely dry well. Oil-Red O was eluted again and absorbance was measured at 490 nm with an ELISA reader.

[Mathematical Expression]

Lipid accumulation (%) = 100 - (A - B) / A × 100

A: Absorbance of the control group

B: absorbance of the sample treatment group

On day 9 of differentiation, the adipocytes were washed with PBS and pulverized with a sonicator for 10 seconds using a homogenizing solution (50 mM Tris, 154 mM KCl, 1 mM EDTA, pH 7.4). The cells were then centrifuged at 3,000 rpm for 5 minutes to obtain cell supernatants. Quantitative colorimetric measurements were made at 550 nm wavelength using neutral fat for quantification.

The results are shown in Figs.

FIG. 8 shows the amount of lipid production (%) according to the addition of the extract of Ganoderma lucidum extract.

FIG. 9 shows the change in the neutral fat amount according to the addition of the extract of Ganoderma lucidum extract.

In conclusion, it was found that the extract and fractions of Ganoderma lucidum have a pancreatic lipase inhibitory activity and can inhibit adipocyte differentiation and lipid metabolism, thereby reducing lipid production and neutral mass, and thus can be effectively used for controlling or improving lipid metabolism .

Claims (5)

A food composition for improving lipid metabolism characterized by containing an extract of Angelica tenuissima Nakai.
A pharmaceutical composition for preventing or treating a lipid metabolism-related disease, which comprises an extract of Angelica tenuissima Nakai.
3. The method according to claim 1 or 2,
Wherein the extraction solvent of the extract is selected from the group consisting of purified water, methanol, ethanol, butanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, hexane and mixtures thereof.
The method according to claim 1,
The composition may comprise,
Wherein the composition is a health functional food in powder, granule, tablet, capsule or beverage form.
3. The method of claim 2,
Wherein said lipid metabolism-related disease is at least one of obesity, diabetes, hypercholesterolemia, hyperlipidemia, hypersomnia, lipoproteinemia or arteriosclerosis.

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