KR20130044707A - Composition for improving metabolism disorder comprising syringaresinol - Google Patents

Abstract

PURPOSE: A composition containing syringaresinol is provided to suppress fatty acid synthesis, to promote fatty acid oxidation, to enhance PGC 1 expression, and to prevent and treat obesity and hyperlipidemia. CONSTITUTION: A composition for preventing or treating dysbolism contains 1-80 wt% of syringaresinol as an active ingredient. Dysbolism includes disorders of carbohydrate metabolism or lipid metabolism, selected from the group consisting of obesity, type 2 diabetes, hyperlipidemia, and fatty liver. Syringaresinol is contained in extracts which is prepared using one or more selected among linseed, Phellodendron amurense Ruprecht, Acanthopanax senticosus, sesame seeds, and ginseng fruits. [Reference numerals] (AA) mRNA expression of relative SIRT 1; (BB) Fat cell; (CC) Liver cell; (DD) Muscle cell

Description

Composition for improving metabolism disorder comprising syringaresinol

The present invention relates to a composition for improving metabolic dysfunction comprising siringarecinol.

The lifestyle changes and overflowing high-calorie foods caused by civilization's selfishness expose humans to the risk of metabolic diseases represented by obesity, type 2 diabetes, hyperlipidemia and fatty liver. Considering that obesity is a major cause of metabolic diseases such as type 2 diabetes, hyperlipidemia, and fatty liver, preventing or improving obesity may prevent other metabolic diseases.

Obesity generally occurs when energy influx increases compared to body energy consumption. Specifically, surplus energy is stored in adipose tissue in the form of triglycerides. Adipose tissue can store large amounts of surplus energy like a rubber balloon, but at the same time increases the size of fat cells. In addition, if the amount of surplus energy that needs to be stored is not adequate for fatty tissue alone, lipid dysregulation, which stores energy in other tissues such as muscle and liver, results in lipotoxicity. In addition, the fat stored in each tissue is converted into free fatty acids through lipolysis, which is known to neutralize the insulin signaling system through signaling mechanisms such as JNK and PKC. Inhibition of the insulin signaling system inevitably leads to insulin resistance, which leads to various metabolic diseases such as hyperglycemia, hyperlipidemia, hypercholesterolemia, type 2 diabetes, fatty liver, and the like. As such, obesity is more serious because it is accompanied by various adult diseases as well as cosmetic problems.

Korean Patent Publication No. 10-2011-0110053 (2011.10.06. Publication)

The present invention is to provide a composition having an excellent metabolic prevention or improvement effect, specifically by controlling the fat metabolism having an excellent metabolic prevention or improvement effect. In another aspect, the present invention is to provide a food and pharmaceutical composition having an excellent metabolic prevention or improvement effect.

One aspect of the present invention provides a composition for preventing or ameliorating metabolic dysfunction, including syringaresinol as an active ingredient.

Another aspect of the present invention provides a food and pharmaceutical composition for preventing or ameliorating metabolic dysfunction, including siringarecinol as an active ingredient.

The composition according to the present invention comprises a siringaresinol as an active ingredient, thereby increasing the expression of SIRT 1 (Sirtuin 1), by inhibiting fatty acid synthesis while promoting fatty acid oxidation, by increasing the expression of PGC 1, It is excellent in preventing and improving metabolic abnormalities, in particular obesity, type 2 diabetes, hyperlipidemia or fatty liver.

Figure 1 is a schematic diagram showing a method for separating and purifying shiringaresinol from ginseng fruit extract.
2 is the chemical structure of siringaresinol.
Figure 3 is a graph showing the expression changes of SIRT 1 gene after the treatment of siringaresinol 20, 50, 100 μM to human adipocytes, hepatocytes and myocytes.
Figure 4 is a graph showing the expression reduction of genes involved in fatty acid synthesis in human adipocytes treated with 50 μM of cyringaresinol.
5 is a graph showing the expression reduction of genes involved in fatty acid synthesis in human hepatocytes treated with 50 μM of siringarecinol.
Figure 6 is a graph showing the expression reduction of genes involved in fatty acid synthesis in human myocytes treated with 50 μM of cyringaresinol.
Figure 7 is a graph showing the increased expression of genes involved in fatty acid oxidation in human adipocytes treated with 50 μM of cyringaresinol.
8 is a graph showing increased expression of genes involved in fatty acid oxidation in human hepatocytes treated with 50 μM of siringarecinol.
Figure 9 is a graph showing the increased expression of genes involved in fatty acid oxidation in human myocytes treated with 50 μM of cyringaresinol.
10 is a graph showing the increase in fatty acid oxidation in human adipocytes, hepatocytes and myocytes treated with 50 μM of siringarecinol.
11 is a graph showing the increased expression of PGC 1a, an energy metabolic regulatory gene, in human adipocytes, hepatocytes and myocytes treated with 50 μM of cyringaresinol.
12 is a graph showing the increased expression of PGC 1b, an energy metabolic regulator gene, in human adipocytes, hepatocytes, and myocytes treated with 50 μM of cyringaresinol.

As used herein, the term "extract" is a broad concept including all materials obtained by extracting the components of natural products, regardless of the extraction method, extraction solvent, extracted components or the form of the extract.

Controlling fat metabolism can prevent over-accumulation of fat, which will prevent and improve obesity. One way to prevent excessive accumulation of fat is dietary restriction, or calorie restriction. Limiting your diet can not only help you lose weight, but also improve your metabolism by improving blood lipid levels. In the case of dietary restriction, the expression of SIRT 1 protein is increased, and if the dietary restriction leads to an increase in the activity of SIRT 1, dietary restriction is similar to that of dietary restriction. have. SIRT 1 is a NAD ⁺ -dependent histone protein deacetylase that regulates energy metabolism such as glucose synthesis, fatty acid oxidation and the like. Therefore, a substance capable of regulating the expression of SIRT 1 may regulate fat metabolism and further prevent or improve metabolic abnormalities such as obesity, type 2 diabetes, hyperlipidemia, and fatty liver.

One aspect of the present invention provides a composition for preventing or ameliorating metabolic dysfunction, including siringaresinol as an active ingredient.

Living things ingest nutrients in order to sustain life, digest it, absorb it into body composition, and store it as an energy source. In addition, it decomposes and performs various biological activities with energy released at that time. Such a phenomenon is called "metabolism". As used herein, the term “metabolism” refers to a state in which a living organism including a human or an animal cannot perform the metabolic activity smoothly, and includes a glucose metabolism abnormality or a lipid metabolism abnormality. Glucose metabolism abnormalities or lipid metabolism abnormalities include symptoms or diseases caused by disorders of glucose or lipid metabolism, in particular those caused by excessive accumulation of fat due to fatty acid metabolism disorders, examples of which include obesity, diabetes mellitus, Hyperlipidemia or fatty liver.

Siringaresinol is a lignan compound having a chemical structure as shown in FIG. 2, which may be obtained through chemical synthesis, or may be extracted from at least one of flaxseed, yellowish white, thorny thorn, sesame and ginseng fruit. The flaxseed, baekbaek, thorns and sesame seeds include all the parts of the plant, for example, the leaves, stems, roots, berries or seeds of each plant, ginseng fruit includes ginseng fruit peel or flesh.

Siringarecinol can increase the expression of SIRT 1, inhibit fatty acid synthesis, and promote oxidation of fatty acids, thereby inhibiting fat accumulation in the body. This may be confirmed by inhibiting expression of fatty acid synthesis related genes such as ADD1 / SREBP1c, ACC, FAS, promoting expression of fatty acid oxidation related genes such as ACO, CPT1, mCAD, and increasing fatty acid oxidation. Can be. In addition, siringarecinol may promote energy metabolism, specifically fat metabolism, thereby increasing body fat consumption and inhibiting the accumulation of excess fat, which is a peroxisome that regulates the expression of genes involved in energy metabolism. increased expression of proliferator-activated receptor coactivator 1. Thus, a composition comprising siringarecinol can prevent or ameliorate metabolic disorders such as obesity, diabetes mellitus, hyperlipidemia or fatty liver due to a glycemic or lipid metabolism disorder.

In one aspect of the invention, the siringaresinol can be obtained by extracting at least one of flaxseed, yellowish white, thorny pomegranate, sesame and ginseng fruit with at least one selected from the group consisting of water, an organic solvent and a mixture of water and an organic solvent. have. The organic solvent includes one or more selected from the group consisting of alcohol, acetone, ether, ethyl acetate, diethyl ether, ethyl methyl ketone and chloroform, but is not limited thereto. The alcohol includes a C ₁ ~ C ₅ lower alcohol, C ₁ ~ C ₅ lower alcohol is one selected from the group consisting of methanol, ethanol, isopropyl alcohol, n-propyl alcohol, n-butanol and isobutanol Including but not limited to the above.

In another aspect of the present invention, siringarecinol may be included in the composition as an extract of flaxseed, yellowish white, thorny creeper, sesame and ginseng fruit, among which fractions may be particularly effective in improving metabolic dysfunction.

In one aspect of the invention, the method of separating and purifying the siringaresinol from ginseng fruit may comprise the following steps. Preparing an alcohol extract of ginseng fruit pulp; Eluting the prepared alcohol extract with a solvent comprising at least one of water and alcohol to obtain a fraction; And performing chromatography, in particular thin film chromatography (TLC), using an organic solvent as the developing solvent for the obtained fraction. The organic solvent includes at least one selected from the group consisting of alcohol, acetone, ether, ethyl acetate, diethyl ether, ethyl methyl ketone, and chloroform, and the alcohol includes C ₁ to C ₅ alcohols. The composition according to one aspect of the present invention may include the purified siringaresinol as an active ingredient as described above.

The composition according to one aspect of the invention comprises from 1% to 80% by weight, in particular from 5% to 60% by weight, more specifically from 10% to 30% by weight, based on the total weight of the composition can do. When included in the above range is not only suitable for showing the intended effect of the present invention, it can satisfy both the stability and safety of the composition, it may be appropriate to use in the above range in terms of cost-effectiveness. Specifically, when less than 1% by weight of shirringarinol, sufficient metabolic improvement effects cannot be obtained, and when it exceeds 80% by weight, safety and formulation stability may be lowered.

One aspect of the present invention provides a food composition comprising siringaresinol as an active ingredient. The food composition may prevent or ameliorate metabolic disorders such as obesity, claim 2 diabetes, hyperlipidemia or fatty liver, and include a favorite food or a health food composition.

The formulation of the food composition is not particularly limited, but may be, for example, formulated into a liquid such as tablets, granules, powders, drinks, caramels, gels, bars, and the like. The food composition of each formulation can be blended with the ingredients commonly used in the field in addition to the active ingredient without difficulty by those skilled in the art depending on the purpose of formulation or use, and synergistic effect can be obtained when the composition is applied simultaneously with other ingredients.

Dosage determination of the active ingredient is within the level of one skilled in the art, the daily dosage of which is for example from 0.1 mg / kg / day to 5000 mg / kg / day, more specifically from 50 mg / kg / day to 500 mg / kg / day, but is not limited thereto, and may vary depending on various factors such as age, health condition, and complications of the subject to be administered.

One aspect of the present invention provides a pharmaceutical composition comprising siringaresinol as an active ingredient. The pharmaceutical composition may prevent or improve metabolic abnormalities, and in particular, may prevent or improve obesity, claim 2 diabetes, hyperlipidemia or fatty liver.

The pharmaceutical composition according to one aspect of the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, and the like.

Formulations for oral administration may be tablets, pills, soft and hard capsules, granules, powders, granules, solutions, emulsions or pellets, but are not limited thereto. It is not. These formulations may contain, in addition to the active ingredient, diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose or glycine), glidants (e.g. silica, talc, stearic acid or polyethylene glycol), or binders (e.g. magnesium aluminum Silicates, starch pastes, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose or polyvinylpyrrolidine). It may optionally contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavors, or sweeteners. The tablets may be prepared by conventional mixing, granulating or coating methods.

Formulations for parenteral administration may be, but are not limited to, injections, drops, lozenges, ointments, gels, creams, suspensions, emulsions, suppositories, patches or spraying agents.

The active ingredient of the pharmaceutical composition according to one aspect of the present invention will vary depending on the age, sex, weight, pathology and severity of the subject to be administered, the route of administration or the judgment of the prescriber. Dosage determination based on these factors is within the level of those skilled in the art, the daily dosage of which is for example 0.1 mg / kg / day to 100 mg / kg / day, more specifically 5 mg / kg / day to 50 mg / kg / day, but is not limited thereto.

One aspect of the present invention provides a cosmetic composition comprising siringaresinol as an active ingredient. The cosmetic composition may prevent or improve metabolic abnormalities, and in particular, may prevent or improve obesity, claim 2 diabetes, hyperlipidemia or fatty liver.

The cosmetic composition according to the present invention may be provided in all formulations suitable for topical application. For example, it may be provided as a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an oil phase in water, a suspension, a solid, a gel, a powder, a paste, a foam or an aerosol composition. Compositions of such formulations may be prepared according to conventional methods in the art.

The cosmetic composition according to the present invention may further include a moisturizer, an emulsifier, a surfactant, a ultraviolet absorbent, a preservative, a fungicide, an antioxidant, a pH adjuster, organic and inorganic pigments, flavors, coolants, or limiting agents. . The blending amount of the above components can be easily selected by those skilled in the art within the range that does not impair the object and effect of the present invention, the blending amount may be 0.01 to 5% by weight, specifically 0.01 to 3% by weight based on the total weight of the composition. have.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to Examples and Experimental Examples. However, these Examples and Experimental Examples are provided only for the purpose of illustration in order to facilitate understanding of the present invention, but the scope and scope of the present invention is not limited thereto.

EXAMPLES Isolation and Analysis of Cyringareresinol

1. Pretreatment of Ginseng Fruit

The raw ginseng fruit was harvested, the seeds were separated and removed, and then the skin of the ginseng fruit was removed, and only the flesh was dried by daylight drying or hot air to prepare a ginseng fruit flesh dried product.

2. Isolation and Analysis of Shilingaresinol from Ginseng Fruit Pulp Extract

Water or alcohol 3L was added to 1 kg of the ginseng fruit pulp dried product prepared above, followed by extraction at room temperature or reflux, followed by filtration and concentration under reduced pressure at 40-45 ° C. to obtain 300 g of ginseng fruit pulp extract. The extract was treated with ether to remove lipid soluble components, crude saponin was extracted with butanol and concentrated. This was separated and purified to obtain syringal lysinol. Specific methods are as follows. First, 194 g of the sample was purified by reversed-phase column chromatography. At this time, 100% water was initially used as the elution solvent, and methanol was gradually increased by 10%, and finally 100% methanol solvent was used. . As a result, GB-1 to GB-10 fractions were obtained. The fractions GB-3 showing SIRT1 expression activity were selected and concentrated. Then, 50% aqueous methanol was used to separate the fractions from the Sephadex LH-20 column Followed by chromatography (Sephadex LH-20 column chromatography). GB-3-6 (3F) showing the SIRT1 expression activity was selected and concentrated. TLC of preparative silica gel was performed using chloroform: methanol (10: 1) as a developing solvent. The active fraction having an Rf value of 0.67 was purified. The separation and purification method is shown schematically in FIG. 1.

NMR spectroscopy and database search were performed to identify the isolated and purified active compounds as syringaresinol. In order to reconfirm this, mass analysis was performed. As a result of measuring ESI-mass in positive mode, [M + Na] + was observed at m / z 440.9 and [2M + Na] + was observed at m / z 858.9. It was found that the molecular weight is 418. In addition, NMR spectroscopic analysis was performed to determine the chemical structure, and the results are shown in FIG. 2. As can be seen in Figure 2, the isolated and purified active compound has the chemical structure of the siringaresinol.

Thus, the shiringaresinol was isolated from the ginseng fruit pulp.

Experimental Example 1 Evaluation of SIRT 1 Expression Promoting Effect in Adipose Cells, Hepatocytes, and Myocytes

In order to evaluate the effect of SIRT 1 gene expression on the adipocyte, hepacyte and myocyte of siringaresinol, the following experiment was performed.

Human adipocytes, hepatocytes and myocytes were purchased from ZENBIO (Research Triangle Park, NC, USA) and adipocyte dedicated media (OM-AM, ZENBIO), hepatocyte dedicated media (HM-2, ZENBIO) and myocytes, respectively. Culture was carried out in a 5% CO ₂ incubator using a dedicated medium (SKM-D, SKM-M, ZENBIO). Each cell was treated with shiringaresinol dissolved in DMSO for 24 hours at 20, 50, 100 μM concentration, and negative control was treated with DMSO with a volume of 1/1000 of the medium.

Cells treated with each sample were washed twice with cold PBS, and RNA was extracted therefrom using Trizol reagent (TRIzol agent, Invitrogen). CDNA was synthesized using a reverse transcription system (Fermentas, Glen Burnie, MD, USA) with 5 mg of RNA extracted and quantified. Expression of each gene was measured by qRT-PCR using primers designed for each gene of cDNA and SIRT 1 and GAPDH. PCR reaction and analysis were performed using a Rotor-Gene 3000 system (Corbett Research, Sydney, Australia). mRNA relative expression results are shown in FIG. 3.

As can be seen in FIG. 3, siringarecinol can increase the expression of SIRT 1 in human adipocytes, hepatocytes and myocytes in a concentration dependent manner. Based on this, it can be seen that siringaresinol may improve metabolic abnormalities, specifically lipid metabolic abnormalities.

Experimental Example 2 Evaluation of Gene Expression Regulation of Adipose Metabolism in Adipose and Hepatocytes

In the same manner as in Experiment 1, human adipocytes, hepatocytes and myocytes were treated with 50 μM siringarecinol, washed with PBS, and RNA was extracted to synthesize cDNA. Changes in expression patterns of fatty acid synthesis-related genes such as ADD1 / SREBP1c, ACC, and FAS and fatty acid oxidation-related genes such as ACO, CPT1, and mCAD were measured using qRT-PCR. 4 to 9 show the results compared with the control treated with only DMSO as a sample.

As can be seen in Figures 4 to 6, siringaresinol inhibited the expression of genes that induce fatty acid synthesis. In addition, as can be seen in Figures 7 to 9, siringaresinol increased the expression of genes that promote fatty acid oxidation. Based on this, it can be seen that siringaresinol can inhibit fat accumulation by inhibiting fat synthesis and increasing fat consumption.

Experimental Example 3 Evaluation of Fatty Acid Oxidation Promoting Capacity

50 μM siringarecinol was treated to human adipocytes, hepatocytes and muscle cells in substantially the same manner as in Experimental Example 1. The cells were washed with PBS and incubated in a medium for fatty acid oxidation measurement for one day, and then the medium was rolled up to measure the amount of ³ H ₂ O contained therein. The result compared with the untreated group is shown in FIG.

As can be seen in FIG. 10, siringarecinol can increase fatty acid oxidation in human adipocytes, hepatocytes and myocytes. Based on this, it can be seen that siringaresinol can inhibit the accumulation of fat in the body.

Example 5 PGC 1 Expression Increasing Effect of Siringaresinol

In a substantially the same manner as in Experimental Example 1, 50 μM siringaresinol was treated to human adipocytes, hepatocytes and myocytes, washed with PBS, and RNA and cDNA were sequentially extracted. MRNA expression of PGC 1α and PGC 1β, which regulate the expression of genes involved in energy metabolism, was analyzed by qRT-PCR using dedicated primers. The expression results of PGC 1α and PGC 1β compared to the control treated with DMSO only as a sample are shown in FIGS. 11 and 12, respectively.

As can be seen in FIGS. 11 and 12, siringarecinol can increase the expression of PGC 1α and PGC 1β in human adipocytes, hepatocytes and myocytes. Based on this, it can be seen that siringaresinol can stimulate energy metabolism and suppress body fat accumulation.

Formulation examples of compositions according to one aspect of the present invention are described below, but may be applied to various other formulations, which are not intended to be limiting but merely illustrative of the invention.

[Formulation Example 1] Health food

Siringalecinol ................................... 1000 mg

Vitamin mixtures

Vitamin A Acetate ............... 70 μg

Vitamin E ....................... 1.0 mg

Vitamin B1 ...................... 0.13 mg

Vitamin B2 ........... 0.15 mg

Vitamin B ........................ 0.5 mg

Vitamin B12 ......... 0.2 μg

Vitamin C ............................ 10 mg

Biotin ............... 10 μg

Nicotinic Acid Amide ... 1.7 mg

Folic acid ......................................... 50 ㎍

Calcium Pantothenate ......................................... 0.5 mg

Mineral mixture

Ferrous Sulfate ........................ 1.75 mg

Zinc Oxide ..................... 0.82 mg

Magnesium Carbonate ......... 25.3 mg

Potassium monophosphate ......................................... 15 mg

Dibasic calcium phosphate ............... 55 mg

Potassium Citrate ............... 90 mg

Calcium Carbonate ..................... 100 mg

Magnesium Chloride ............ 24.8 mg

Although the composition ratio of the said vitamin and mineral mixture was mixed and consisted with the component suitable for a healthy food in a preferable Example, the compounding ratio may be arbitrarily modified.

[Formulation Example 2] Health drinks

Siringaresinol ..................... 1000 mg

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

Oligosaccharide ......................... 100 g

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

Purified water ..................

The above components are mixed according to a conventional health drink manufacturing method, and the mixture is stirred and heated at 85 DEG C for about 1 hour, and then the solution is sterilized by filtration.

Formulation Example 3 Tablet

100 mg of siringaresinol, 50 mg of soy extract, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch and 4 mg of magnesium stearate were added and 40 mg of 30% ethanol was added to form granules. Dry and tablet into tablets using a tablet press.

[Formulation Example 4]

100 mg of siringaresinol, 50 mg of soy extract, 100 mg of glucose, and 600 mg of starch are mixed and 100 mg of 30% ethanol is added to form granules, followed by drying at 60 ° C. to form granules, followed by filling into fabrics. .

Formulation Example 5 Cream

A cream is prepared according to a conventional method with the composition described in the table below.

ingredient Content (% by weight) Siringaresinol 3.0 Polyethylene Glycol Monostearate 2.0 Glycerin Monostearate 5.0 Cetyl alcohol 4.0 Squalene 3.0 Tri-2-ethylhexane glyceryl 6.0 Sphingolipid 1.0 1,3-butylene glycol 7.0 Purified water Balance

Claims (8)

Composition for preventing or ameliorating metabolic dysfunction comprising siringaresinol as an active ingredient.
The method of claim 1,
Metabolic dysfunction is a composition comprising glucose metabolic dysfunction or lipid metabolism dysfunction.
3. The method of claim 2,
Carbohydrate metabolic dysfunction or lipid metabolic dysfunction comprises at least one selected from the group consisting of obesity, type 2 diabetes, hyperlipidemia and fatty liver.
The method of claim 1,
Siringaresinol is a composition comprising one or more extracts selected from flaxseed, yellowish white, bramble, sesame and ginseng fruit.
The method of claim 1,
The composition comprises 1% to 80% by weight of siringaresinol based on the total weight of the composition.
The method of claim 1,
The composition inhibits fatty acid synthesis and promotes fatty acid oxidation.
7. The method according to any one of claims 1 to 6,
The composition is a food composition.
7. The method according to any one of claims 1 to 6,
The composition is a pharmaceutical composition.

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