KR20220034688A - A composition for preventing or treating obesity and atherosclerosis comprising Galla Rhois ethanol extract as active ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing or treating obesity and arteriosclerosis comprising an extract of Rhus chinensis as an active component, wherein the extract of Rhus chinensis extracted with water, an organic solvent, or a mixed solvent thereof regulates the expression of genes involved in lipid accumulation and lipid metabolism, suppresses the increase in the size of body fat caused by a high-fat diet, suppresses increase in blood cholesterol and triglycerides, and regulates the expression of genes related to arteriosclerosis, such as cell adhesion substances and apolipoprotein, in vascular smooth muscle cells, so that it is possible to be used as a very effective pharmaceutical composition for preventing or treating obesity, hyperlipidemia, and arteriosclerosis related to the same.

Description

A pharmaceutical composition for preventing or treating obesity and arteriosclerosis comprising an ethanol extract of baeja baeja as an active ingredient {A composition for preventing or treating obesity and atherosclerosis comprising Galla Rhois ethanol extract as active ingredient}

The present invention relates to a composition for preventing, improving, or treating obesity and arteriosclerosis, comprising as an active ingredient of an aqueous solution of pentagram lower alcohol solution.

Recently, as well as in developed countries, circulatory diseases such as arteriosclerosis, cerebral hemorrhage, hypertension, heart disease, stroke, and cerebral infarction are the leading causes of death in Korea, along with an increase in the obese population due to excessive nutritional intake due to westernization of diet. These adult diseases are occurring frequently among middle-aged and elderly people due to changes in dietary habits and internal and external environmental stress as they develop into modern society.

Arteriosclerosis is a disease in which fat accumulates on the walls of blood vessels over a long period of time and thickens the walls of blood vessels. Hyperlipidemia, especially hypercholesterolemia, is a major risk factor, and hypertension and diabetes are also known risk factors. Recently, the incidence of arteriosclerosis and related cerebro-cardiovascular diseases has been greatly increased not only in developed countries, but also in the increase of the obese population due to excessive nutritional intake due to the westernization of diet and the aging of the population. When looking at the progress of arteriosclerosis, damage to the vascular endothelial cells composing the inner surface of blood vessels due to oxidized lipids, high blood pressure, and rapid blood flow occurs first, and then fat gradually accumulates in the vessel wall, leading to infiltration of macrophages and T cells, etc. The inflammatory reaction proceeds and the vessel wall thickens due to proliferation of vascular smooth muscle cells. As a result, the blood vessels become narrow, making the flow of blood difficult.

After low-density lipoprotein (LDL) in the blood accumulates above a threshold in the extracellular matrix of the endothelial tissue in conjunction with various risk factors, the accumulated lipoprotein is mildly oxidized by oxidizing substances in the tissue, which It acts on endothelial cells and increases the expression of attachment substances (VCAM-1, ICAM-1, E-selectin) required for monocytes in the blood to attach and move to the subendothelial tissue, thereby allowing monocytes to flow in. The introduced monocytes are differentiated into macrophages and, while activated, promote the influx of monocytes by stimulation of oxidized lipids, substances that proliferate cells (M-CSF, G/M-CSF, PDGF), and cytokines involved in inflammation (IL-1, TNF-a), secretes tissue factor that promotes coagulation, and also secretes oxidizing substances to further oxidize lipids to make oxidized lipids,36 Ingesting large amounts of oxidized lipids on their own through scavenger receptors to reduce lipids It differentiates into deposited foam cells.

Atherosclerosis begins with damage to vascular endothelial cells, and when endothelial cells become dysfunctional or endothelial cells undergo an inflammatory response, vascular cell adhesion molecules (VCAM)- 1, Intercellular adhesion molecule (ICAM)-1 and E-selectin are overexpressed, and immune cells of lymphocytes, polymorphonuclear leukocytes and monocytes in the blood, platelets, lipids, etc. attach to them, thereby permeating into tissues this will increase Afterwards, the immune cells secrete growth factors, chemotactic factors, and inflammatory cytokines (interleukin-1 and tumor necrosis factor (TNF)-a, etc. Atherosclerotic lesions develop (Libby, P., Nature, 420(6917), 868-874, 2002; Plutzky, J., et al., Journal of Diabetes and its Complications, 16(6), 501-415). , 2002; Libby, P., American Journal of Clinical Nutrition, 83(2), 456s-460s, 2006).

Apolipoprotein is a component of lipoprotein, a complex that transports fat in the blood, and provides a structural skeleton of lipoprotein and plays a role in masking hydrophilic fat. Also, apolipoproteins A1 and B are known as major risk factors for atherosclerosis (Korean Society of Laboratory Medicine, 2012; Bennet, Annam et al. Archives of internal medicine, 168(6), 598-608, 2008).

Currently, treatment for arteriosclerosis is focused only on preventing and suppressing the occurrence of arteriosclerosis by lowering the blood cholesterol level, and treatment to stabilize or reduce the arteriosclerosis that has already occurred has not been implemented so far. Drugs known to have anti-inflammatory and anti-lipid effects have a problem in that they are difficult to apply in clinical practice because they require several times or tens of times the dose allowed for humans for their effects.

For the improvement of blood circulation and prevention or treatment of thrombotic diseases, a lot of research is being conducted on pharmaceutical materials or health functional food materials that can inhibit the formation of blood clots and effectively control the concentration of cholesterol and triglycerides in the blood.

Korean Patent Publication No. 10-2020-0084212

Accordingly, the present inventors reduced food intake and body weight as a result of administration of the baeja extract to mice bred on a high-fat diet while conducting a study on the anti-obesity efficacy of the extract, reducing the weight of adipose tissue, and reducing the lipids in the serum and total cholesterol, and suppressing the expression of TNF-a-induced cell adhesion material and apolipoprotein in vascular smooth muscle cells, thereby completing the present invention.

Accordingly, one aspect of the present invention is to provide a composition for preventing, improving, or treating obesity comprising the extract of baeja baeja as an active ingredient.

One aspect of the present invention is to provide a composition for the prevention, improvement or treatment of cardiovascular diseases comprising the extract of baekjae baeja as an active ingredient. The cardiovascular disease includes hypercholesterolemia, hyperlipidemia, arteriosclerosis, atherosclerosis, peripheral vascular disease, dyslipidemia, hyperbetalipoproteinemia, hypoalphalipoproteinemia, hypercholesterolemia, hypertriglyceridemia, familial hyperlipidemia Cholesterolemia, Cardiovascular Disorders, Coronary Heart Disease, Coronary Artery Disease, Coronary Vascular Disease, Angina, Ischemia, Cardiac Ischemia, Thrombosis, Myocardial Infarction, Stroke, Peripheral Vascular Disease, Reperfusion Injury, Restenosis after Angioplasty, Hypertension, Congestive Heart failure, diabetes mellitus, diabetic vascular complications, obesity and endotoxemia may be present.

One aspect of the present invention is to provide an anti-inflammatory composition comprising the extract of baeja baeja as an active ingredient.

One aspect of the present invention is to provide a composition for the prevention, improvement or treatment of cancer comprising the extract of baekjaeja as an active ingredient.

Another aspect of the present invention is to provide a method for preparing a composition for preventing, improving or treating obesity.

Another aspect of the present invention is to provide a method for preparing a composition for preventing, improving or treating the cardiovascular disease.

Another aspect of the present invention is to provide a health functional food composition comprising the composition.

Another aspect of the present invention is to provide a pharmaceutical composition comprising the composition.

Another aspect of the present invention is to provide a cosmetic composition comprising the composition.

One aspect of the present invention provides a composition for preventing, improving or treating obesity comprising an extract of Galla Rhois as an active ingredient.

In one aspect of the present invention, the active ingredient provides a composition for preventing, improving or treating obesity, which has the effect of reducing adipose tissue weight.

One aspect of the present invention provides an anti-inflammatory composition comprising an extract of Galla Rhois as an active ingredient.

One aspect of the present invention provides a composition for preventing, improving or treating cancer comprising an extract of Galla Rhois as an active ingredient.

In one aspect, the present invention provides a composition for preventing, improving or treating cardiovascular disease comprising an extract of Galla Rhois as an active ingredient, wherein the cardiovascular disease is selected from diseases consisting of hypercholesterolemia, hyperlipidemia and arteriosclerosis. Any one or more, it provides a composition for the prevention, improvement or treatment of cardiovascular disease.

In one aspect of the present invention, the active ingredient provides a composition for the prevention, improvement or treatment of cardiovascular disease, which has an effect of reducing blood cholesterol.

In one aspect of the present invention, the active ingredient provides a composition for preventing, improving or treating cardiovascular diseases, wherein the active ingredient inhibits the expression of TNF-a-induced cell adhesion material and apolipoprotein in vascular smooth muscle cells.

In one aspect of the present invention, the baeja is a worm nest formed by the stab wound of the quince aphid ( Melaphis chinensis Bell) on the leaves of Rhus javanica L. or Sumacaceae (Anacardiaceae), it provides a composition.

In one aspect of the present invention, the extract is an extract extracted using water, C ₁ -C ₅ alcohol, or C ₁ -C ₅ alcohol aqueous solution, it provides a composition.

In one embodiment, the concentration of the C ₁ -C ₅ alcohol aqueous solution is 45% to 95% (v / v), it provides a composition.

In one embodiment, the extract provides an extract of 75% to 85% (v/v) of an aqueous ethanol solution, the composition.

In one aspect of the present invention, the composition provides a health functional food, composition.

In one aspect of the present invention, the composition provides a pharmaceutical composition, the composition.

In one aspect of the present invention, the composition provides a cosmetic composition, the composition.

In one embodiment, the pharmaceutical composition provides a composition formulated in any one of injections, powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and external preparations.

In the present technology, circulatory system diseases and obesity, including hyperlipidemia, are increasing due to the increase in lipid intake due to the westernization of diet, and there is a need for a therapeutic agent for the prevention and treatment of these diseases. It can be usefully used for preventing and treating arteriosclerosis or atherosclerotic diseases by inhibiting and reducing the expression of intravascular adhesion substances in vascular endothelial cells to inhibit thickening of the vessel wall.

Diseases or disorders that can be treated, prevented, or ameliorated by the composition comprising the extract of the pear extract obtained in the present invention are specifically hypercholesterolemia, hyperlipidemia, arteriosclerosis, atherosclerosis, peripheral vascular disease, dyslipidemia, Hyperbetalipoproteinemia, hypoalphalipoproteinemia, hypercholesterolemia, hypertriglyceridemia, familial hypercholesterolemia, cardiovascular disorder, coronary heart disease, coronary artery disease, coronary artery disease, angina pectoris, ischemia, cardiac ischemia, thrombosis , myocardial infarction, stroke, peripheral vascular disease, reperfusion injury, restenosis after angioplasty, hypertension, congestive heart failure, diabetes mellitus, diabetic vascular complications, obesity and endotoxemia.

The composition containing the extract obtained from the present invention is excellent in anti-inflammatory effect, the effect is excellent as a cosmetic composition for anti-inflammatory use.

The effects of the present invention are not limited to the above-mentioned effects. It should be understood that the effects of the present invention include all effects that can be inferred from the following description.

Figure 1 shows the anti-obesity efficacy of the extract of the present invention for each ethanol concentration of baekjae baeja.
Figure 2 shows the anti-obesity efficacy of the 80% ethanol extract of the present invention.
Figures 3a to 3i show changes in the expression of lipid metabolism-related genes of the 80% ethanol extract of pentagram of the present invention.
Figures 4a to 4h show the results of an animal test for obesity prevention of 80% ethanol extract of baekjae baeja of the present invention.
Figure 5 shows the tissue changes of the 80% ethanol extract of Obaeja in the obesity prevention animal model of the present invention.
Figures 6a to 6h shows the results of the animal test for obesity treatment of 80% ethanol extract of pentagram of the present invention.
Figure 7 shows the tissue changes of 80% ethanol extract of Obaeja in the obesity treatment animal model of the present invention.
Figures 8a to 8e show the cell adhesion material, cytokine, apolipoprotein expression inhibitory efficacy of the 80% ethanol extract of baekjae of the present invention.
Figure 9 shows the effect of inhibiting the expression of VCAM-1, NF-κB of the 80% ethanol extract of the present invention.

Unless otherwise specified, all numbers, values, and/or expressions expressing ingredients, reaction conditions, and amounts of ingredients used herein refer to a variety of measures that may occur in obtaining such values, among others, in which such numbers are inherently different. Since they are approximations reflecting uncertainty, it should be understood as being modified by the term “about” in all cases. Also, where the disclosure discloses numerical ranges, such ranges are continuous and inclusive of all values from the minimum to the maximum inclusive of the range, unless otherwise indicated. Furthermore, when such ranges refer to integers, all integers inclusive from the minimum to the maximum inclusive are included, unless otherwise indicated.

In this specification, when a range is described for a variable, the variable will be understood to include all values within the stated range including the stated endpoints of the range. For example, a range of “5 to 10” includes the values of 5, 6, 7, 8, 9, and 10, as well as any subranges such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, etc. It will be understood to include any value between integers that are appropriate for the scope of the recited range, such as 5.5, 6.5, 7.5, 5.5 to 8.5 and 6.5 to 9, and the like. Also for example, ranges from "10% to 30%" include values of 10%, 11%, 12%, 13%, etc. and all integers up to and including 30%, as well as 10% to 15%, 12% to It will be understood to include any subranges such as 18%, 20% to 30%, etc., as well as any value between reasonable integers within the scope of the recited ranges, such as 10.5%, 15.5%, 25.5%, and the like.

In addition, unless otherwise defined, terms and abbreviations used herein may be interpreted as meanings commonly understood by those skilled in the art to which the present invention pertains.

Hereinafter, the invention will be described in detail.

An object of the present invention is to provide a food and pharmaceutical composition containing a composition for preventing and treating obesity and arteriosclerosis, which are one of lifestyle-related diseases. In addition, it is another object of the present invention to provide a composition for preventing, improving or treating cardiovascular diseases including arteriosclerosis.

The present inventors, as a result of intensive research to achieve the above object, found that the composition extracted by organic solvent extraction of oysteriasis not only has an anti-obesity action in vitro, but also an anti-obesity action in experimental animals, based on this finding The present invention has been completed.

One aspect of the present invention provides the following.

(1) A composition for preventing obesity and arteriosclerosis, which can be obtained by extraction with an organic solvent from quince.

(2) The composition of the above (1), wherein the baeja is a worm nest formed by a stab wound of the quince aphid ( Melaphis chinensis Bell) on the leaves of Rhus javanica L. or other faunal plants, Anacardiaceae.

(3) The composition according to (1) or (2) above, wherein the organic solvent extraction comprises extraction with a lower alcohol from pentagram.

(4) The composition according to (3), wherein the lower alcohol is ethanol or methanol.

(5) A pharmaceutical composition comprising the composition for preventing and treating obesity and arteriosclerosis according to any one of (1) to (4) above.

Hereinafter, various aspects of the present invention will be described.

One aspect of the present invention provides a composition for preventing, improving or treating obesity comprising an extract of Galla Rhois as an active ingredient.

As used herein, the term "extract" includes all substances obtained by extracting components from natural products, regardless of the extraction method or type of components. For example, it is a broad concept that includes both extracts of components soluble in solvents from natural products using water or organic solvents, and those obtained by extracting only specific components of natural products. In one embodiment of the present invention, the organic solvent is not particularly limited, and C ₁ to C ₅ lower alcohols such as methanol, ethanol, isopropyl alcohol, n-propyl alcohol, n-butanol and isobutanol, glycerol , polyhydric alcohols such as ethylene glycol, propylene glycol, 1,3-butylene glycol, hydrocarbon solvents such as methyl acetate, ethyl acetate, benzene, n-hexane, diethyl ether, dichloromethane, chloroform, and petroleum It may be a non-polar organic solvent such as ether, methyl acetate, benzene, hexane, chloroform, methylene chloride, dimethyl ether, and ethyl acetate.

In one aspect of the present invention, the active ingredient provides a composition for preventing, improving or treating obesity, which has the effect of reducing adipose tissue weight.

One aspect of the present invention provides an anti-inflammatory composition comprising an extract of Galla Rhois as an active ingredient. In one embodiment, the active ingredient exhibits anti-inflammatory efficacy through the effect of inhibiting inflammatory cytokines as supported in FIG. 8 of the present invention.

One aspect of the present invention provides a composition for preventing, improving or treating cancer comprising an extract of Galla Rhois as an active ingredient.

In one aspect, the present invention provides a composition for preventing, improving or treating cardiovascular disease comprising an extract of Galla Rhois as an active ingredient, wherein the cardiovascular disease is selected from diseases consisting of hypercholesterolemia, hyperlipidemia and arteriosclerosis. Any one or more, it provides a composition for the prevention, improvement or treatment of cardiovascular disease.

In one embodiment, the cardiovascular disease is hypercholesterolemia, hyperlipidemia, arteriosclerosis, atherosclerosis, peripheral vascular disease, dyslipidemia, hyperbetalipoproteinemia, hypoalphalipoproteinemia, hypercholesterolemia, hypercholesterolemia Triglyceridemia, familial hypercholesterolemia, cardiovascular disorders, coronary heart disease, coronary artery disease, coronary artery disease, angina pectoris, ischemia, cardiac ischemia, thrombosis, myocardial infarction, stroke, peripheral vascular disease, reperfusion injury, reperfusion after angioplasty Stenosis, hypertension, congestive heart failure, diabetes mellitus, diabetic vascular complications, obesity and any one or more selected from diseases consisting of endotoxemia, a composition is provided.

In one aspect of the present invention, the active ingredient provides a composition for the prevention, improvement or treatment of cardiovascular disease, which has an effect of reducing blood cholesterol.

In one aspect of the present invention, the active ingredient provides a composition for preventing, improving or treating cardiovascular diseases, wherein the active ingredient inhibits the expression of TNF-a-induced cell adhesion material and apolipoprotein in vascular smooth muscle cells.

In one aspect of the present invention, the baeja is a worm nest formed by the stab wound of the quince aphid ( Melaphis chinensis Bell) on the leaves of Rhus javanica L. or Sumacaceae (Anacardiaceae), it provides a composition.

In one aspect of the present invention, the extract is an extract extracted using water, C ₁ -C ₅ alcohol, or C ₁ -C ₅ alcohol aqueous solution, it provides a composition.

In one embodiment, the concentration of the C ₁ -C ₅ alcohol aqueous solution is 45% to 95% (v / v), it provides a composition.

In one embodiment, the concentration of the C ₁ -C ₅ alcohol aqueous solution is 70% to 90% (v / v), it provides a composition.

In one embodiment, the extract provides an extract of 75% to 85% (v/v) of an aqueous ethanol solution, the composition.

In one aspect of the present invention, the baeja extract may be included in an amount of 0.001 to 90% by weight based on the total weight of the composition. In one embodiment, the extract based on the total weight of the composition is 0.001% by weight or more, 0.01% by weight or more, 0.1% by weight or more, 1% by weight or more, 1.1% by weight or more, 1.5% by weight or more, 2% by weight or more, 3 wt% or more, 5 wt% or more, 10 wt% or more, 20 wt% or more, or 30 wt% or more. In addition, the ginseng flower stem extract is 90% by weight or less, 85% by weight or less, 80% by weight or less, 70% by weight or less, 50% by weight or less, 40% by weight or less, 30% by weight or less, 20% by weight or less, based on the total weight of the composition weight % or less, 10 wt% or less, 5 wt% or less, 4 wt% or less, 3 wt% or less, 2 wt% or less, 1 wt% or less, 0.1 wt% or less, or 0.05 wt% or less.

In one aspect of the present invention, the composition provides a health functional food, composition.

The health food composition of the present invention is to include an extract of baekjae baeja or a solvent fraction fractionated therefrom, and there is no particular limitation on the type thereof. Examples of the food include drinks, meat, sausage, bread, biscuits, rice cakes, sun food, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products. As an active ingredient, the extract of baekjaeja or a solvent fraction fractionated therefrom may be added to food as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method. The effective content may be appropriately determined depending on the purpose of use (for prevention or improvement), and may be included in the range of 0.001 to 70% by weight based on the total weight of the health food. However, in the case of long-term intake for health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. For example, in the case of manufacturing as a health drink, natural carbohydrates or flavoring agents may be included as additives commonly used in the manufacture of beverages in addition to the active ingredient. The natural carbohydrates are conventional such as monosaccharides (eg glucose, fructose, etc.), disaccharides (eg maltose, sucrose, etc.) and polysaccharides (eg dextrin, cyclodextrin, etc.). Phosphorus sugar and sugar alcohols such as xylitol, sorbitol, and erythritol may be included. The natural carbohydrate may be contained in an amount of 1 to 20% by weight, preferably 5 to 10% by weight, based on the total weight of the health food. The flavoring agent may include natural flavoring agents (taumartin, stevia extract, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.). In addition, various nutrients, vitamins, minerals (electrolytes), flavoring agents (synthetic or natural flavoring agents), coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives , glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like. It may also contain fruit flesh for the production of natural fruit juice and fruit juice beverages and vegetable beverages. Although there is no particular limitation on the content of these additives, it may be included in the range of 0.1 to 20% by weight based on the total weight of the health food.

In one aspect of the present invention, the composition provides a pharmaceutical composition, the composition.

In one embodiment, the pharmaceutical composition provides a composition formulated in any one of injections, powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and external preparations.

The pharmaceutical composition of the present invention can be prepared in the form of a pharmaceutical formulation suitable for oral or parenteral administration by further including an appropriate carrier, excipient and/or diluent commonly used in the manufacture of pharmaceuticals. In addition, a pharmaceutical formulation can be prepared according to a conventional method using the pharmaceutical composition of the present invention. In preparing the formulation, the active ingredient may be mixed with a carrier, diluted with a carrier, or enclosed in a carrier in the form of a capsule, sachet or other container. Accordingly, the dosage form is a tablet, pill, powder, capsule, sachet, elixir, suspension, emulsion, solution, syrup, aerosol, soft or hard gelatin capsule, solution or suspension for injection, ointment, cream, gel or lotion. It may be in the form of

Suitable carriers, excipients and diluents that may be included in the pharmaceutical compositions of the present invention are, for example, lactose, dextrose, sucrose, sorbitol, mannitol, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone. , water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, fillers, anti-aggregants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc. which are commonly used in the preparation of formulations may be additionally included. The pharmaceutical composition of the present invention may be formulated using methods well known in the art to provide rapid, sustained or delayed release of an active ingredient after administration to a mammal.

The route of administration of the pharmaceutical composition according to the present invention is not limited thereto, but for example, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, enteral, Sublingual or topical administration is possible.

The dosage of the pharmaceutical composition of the present invention varies depending on the patient's condition and body weight, the degree of disease, drug form, administration route and period, but may be appropriately selected by those skilled in the art. The active ingredient relative to the patient's body weight may be in the range of 0.001 mg/kg to 500 mg/kg, and it is preferable to administer it at 0.001 to 200 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. The above dosage does not limit the scope of the present invention in any way.

In one aspect of the present invention, the composition provides a cosmetic composition, the composition.

Hereinafter, the present invention will be described in more detail through specific examples, experimental examples and formulation examples. The following Examples, Experimental Examples, and Formulation Examples are merely illustrative to aid the understanding of the present invention, and the scope of the present invention is not limited thereto.

Example 1: Preparation of the extract of baekjae

After purchasing five baeja from Gyeongdong Market and drying it, the dried five baeja were dried in the shade, treated with each ethanol concentration, and extracted 2 to 5 times at room temperature, and extracts for each ethanol concentration were obtained. Ethanol concentration is a water solvent without ethanol (0%), 10% ethanol aqueous solution, 20% ethanol aqueous solution, 30% ethanol aqueous solution, 40% ethanol aqueous solution, 50% ethanol aqueous solution, 60% ethanol aqueous solution, 70% ethanol aqueous solution, Extraction was performed with 80% ethanol aqueous solution, 90% ethanol aqueous solution, and 100% ethanol aqueous solution.

Experimental Example 1: Anti-obesity efficacy of extracts by ethanol concentration

3T3-L1 cells, which are pre-adipocytes, are seeded in a 96-well plate at 1 X 10 ⁴ cells/well and incubated until the cells become 100% confluent. When the cells become 100% confluent, culture for 2 more days and then replace with DMEM medium containing 1 μdexamethasone, 0.1 mM 3-isobuty-1-methylxathine, 10 μg/mL insulin, 1% penicillin/Streptomycin, and 10% FBS. Extracts by ethanol concentration (100 μ treatment) were washed once with PBS after 2 days, exchanged with DMEM medium containing 10% FBS, and cultured for a total of 8 days. After induction of differentiation into adipocytes, washed with PBS and 4% formaldehyde was treated and fixed at room temperature for 5 hours, washed three times with PBS, treated with AdiopRed™ assay reagent, stained at room temperature for 10 minutes, and then the lipodifferentiation rate was measured with a Fluorometer.

The anti-obesity efficacy of the extracts for each ethanol concentration was shown in FIG. 1 .

As shown in FIG. 1 , the anti-obesity efficacy was excellent at 50%, 60%, 70%, 80%, and 90% ethanol concentrations compared to other extraction concentrations. In particular, it was the best at 80% ethanol concentration.

The present invention selected an 80% ethanol extract of baeja baeja based on the results of FIG. 1 , and FIG. 2 shows the anti-obesity effect of the ethanol extract of baeja baeja 80%.

As shown in FIG. 2, it was confirmed that the extract of baekjaeja inhibited the accumulation of lipid droplets (green).

Experimental Example 2: Real-time PCR

3T3-L1 pre-adipocytes were cultured in DMEM medium containing 10% BCS and antibiotics, and in an incubator maintained at 37°C with the supply of 5% carbon dioxide. 1 X 10 ⁵ cells/mL of 3T3-L1 cells were aliquoted in a 6 well plate and cultured until the cells were 100% confluent. To differentiate into adipocytes, MDI (0.1 mM 3-isobuty-1-methylxathine, 10 μg/mL insulin, 1 μdexamethasone) was treated with the medium, and the five baeja extracts were treated at concentrations of 0.8, 4, and 20 μ, respectively. After two days, the DMEM medium was replaced with a medium containing FBS, and the extracts of A. baeja extract were treated at concentrations of 0.8, 4, and 20 μg/mL. Cultured for a total of 8 days. After 8 days, remove the medium, wash with PBS, add 1 mL of Trizol to dissolve the cells, transfer to a 1.5 mL microtube, leave at room temperature for 5 minutes, add 200 μ of Cholroform, mix, and leave at room temperature for 10 minutes, then 13,000 After centrifugation at xg, 4℃ for 15 minutes, transfer only the supernatant to a new microtube, add 500 μ of isopropanol, centrifuge at 13,000xg, 4℃ for 15 minutes, and discard the supernatant. After adding 1 mL of 80% EtOH, centrifuge at 13,000xg, 4°C for 5 minutes, discard the supernatant, dry until EtOH is completely gone, and then add 20 μ of RNase free water to dissolve RNA. cDNA was synthesized using PrimeScript Ⅱ 1st strand cDNA synthesis kit. Real-time PCR primers were C/EBPa, C/EBPß, PPARγ, aP2, CD36, LPL, ACC, FAS, HMGCR, and ß-actin. Polymerase was activated by preincubation at 95 °C for 10 minutes, followed by repeating at 95 °C for 30 seconds, 55 °C for 1 minute, and at 72 °C for 30 seconds, a total of 40 times. To quantify the amount of primer for each cDNA sample, it was standardized with β-actin, a housekeeping gene. Primer sequences used for real-time PCR are shown in Table 1 below.

primer order C/EBPa forward primer 5'-ATTTTCTGGGGCAGGAAGTT-3' C/EBPa reverse primer 5'-ACGTCAGAACAACCGAATCC-3' C/EBPß forward primer 5'-AGVAVVTVVVVAVVTACTTT-3' C/EBPß reverse primer 5'-AGCTTGCACGACCCTTCTAA-3' PPARγ forward primer 5'-CTCCAGGTGAACCAAACAACA-3' PPARγ reverse primer 5'-TGACAACTGCAGGATGCATT-3' aP2 forward primer 5'-ACTGCAGGTCTGGCTCAGGA-3' aP2 reverse primer 5'-CAGGTGGCCACAGCCAATAA-3' CD36 forward primer 5'-GCATCCACGTGTTGGCTCAG-3' CD36 reverse primer 5'-TGGACCCATTCCTTCTTGGG-3' LPL forward primer 5'-ATCGGAGAACTGCTCATGATGA-3' LPL reverse primer 5'-CGGATCCTCTCGATGACGAA-3' ACC forward primer 5'-GAATCTCCTGGTGACAATGCTTATT-3' ACC reverse primer 5'-GGTCTTGCTGAGTTGGGTTAGCT-3' FAS forward primer 5'-CTGAGATCCCAGCACTTCTTGA-3' FAS reverse primer 5'-GCCTCCGAAGCCAAATGAG-3' HMGCR forward primer 5'-CATGCAGATTCTGGCAGTCAGT-3' HMGCR reverse primer 5'-CGGCTTCACAAACCACAGTCT-3' ß-actin forward primer 5'-CGCTGATGCATGCCTATGA-3' ß-actin reverse primer 5'-AGAGGTCCACAGAGCTGATTCC-3'

The measurement result is shown in FIG. 3 . (FIGS. 3a to 3i)

As can be seen from Figures 3a to 3i, the expression rate of genes involved in lipid metabolism by the composition showed a tendency to decrease. This suggests that the extract of baekjaeja may have the effect of inhibiting fat accumulation by acting on genes involved in lipid metabolism.

Experimental Example 3: Anti-obesity animal experiment

This experiment is one of the methods to find out the anti-obesity effect of 80% ethanol extract of Obaeja. Experimental animals were purchased 5-week-old C57BL/6 mouse (Central Experimental Animal, Seoul) and acclimatized in a kennel maintained at 50% humidity and 24-26℃ for 1 week, and feed and water were supplied so that they could freely consume it. . The experiment was carried out in two models to confirm the obesity suppression and therapeutic effect of the 80% ethanol aqueous solution extract of Obaeja. Body weight and feed intake were measured once a week. After completion of the experiment, blood was collected and centrifuged at 2,000 x g for 15 minutes, and plasma was collected to measure glucose, leptin, total-cholesterol, and triglyceride.

Experimental models were tested with a preventive model and a treatment model, and are as follows.

<Preventive model>

C57BL/6 mice (5 week-old, Male) were acclimated for 1 week.

The groups were as follows.

NDC: Normal diet control

HFDC : High-fat diet (HFD) control

OS10 : HFD + Orlistat 10 mg/kg

OS20 : HFD + Orlistat 20 mg/kg

GRE20 : HFD + Galla Rhois Ex. 20 mg/kg

GRE40 : HFD + Galla Rhois Ex. 40 mg/kg

GRE80 : HFD + Galla Rhois Ex. 80 mg/kg

In the preventive model, obesity was induced by a high-fat diet, and at the same time, GR extract and orlistat were provided for 10 weeks.

<Treatment model>

C57BL/6 mice (5 week-old, Male) were acclimated for 1 week, and HFD-induced obese mice were induced for 6 weeks.

The groups were as follows.

NDC: Normal diet control

HFDC : High-fat diet (HFD) control

OS10 : HFD + Orlistat 10 mg/kg

OS20 : HFD + Orlistat 20 mg/kg

GRE20 : HFD + Galla Rhois Ex. 20 mg/kg

GRE40 : HFD + Galla Rhois Ex. 40 mg/kg

GRE80 : HFD + Galla Rhois Ex. 80 mg/kg

In the treatment model, obesity was induced for 6 weeks with a high-fat diet, and at the same time, GR extract and orlistat were provided for 10 weeks.

The measurement results are shown in FIGS. 4 to 7 .

As shown in Figs. 4a to 4h, the baeja extract showed an anti-obesity effect by inhibiting the increase in body weight, weight gain inhibition rate, epididymal fat, and liver weight in obesity-induced mice. In addition, the extract of baeja baeja inhibited the increase of triglycerides, cholesterol, glucose, and leptin in obesity-induced mice, thereby showing an anti-obesity effect.

As shown in FIG. 5 , the baeja extract showed an obesity prevention effect by inhibiting the increase in fat size in liver and epididymal adipose tissue in obesity-induced rats.

As shown in FIGS. 6A to 6H , the baeja extract showed an effect of treating obesity by inhibiting the increase in body weight, weight gain inhibition rate, epididymal fat, and liver weight in obesity-induced mice. In addition, the extract of baeja baeja suppressed the increase of triglycerides, cholesterol, glucose, and leptin in obesity-induced rats, thereby showing an anti-obesity effect.

As shown in FIG. 7 , the baeja extract reduced the lipid accumulation in the liver tissue increased in obesity-induced mice, thereby showing an obesity treatment effect. In addition, it was confirmed that the size of epididymal fat was also decreased in the extract of baeja baeja, which was effective in treating obesity.

Experimental Example 4: Confirmation of the effect of inhibiting the expression of cell adhesion substances of the extract

In order to confirm the expression change of the cell adhesion material, the extract prepared in Example 1 was treated with 25, 50, 100 μg/mL in MOVAS (mouse vascular endothelial cell) and TNF using real time PCR and western blot. -a-induced cell adhesion substances, cytokines, and the inhibitory efficacy of apolipoprotein expression was measured. Primer sequences used for real-time PCR are shown in Table 2 below.

primer order VCAM-1 forward primer 5'-ATTTTCTGGGGCAGGAAGTT-3' VCAM-1 reverse primer 5'-ACGTCAGAACAACCGAATCC-3' ICAM-1 forward primer 5'-AGVAVVTVVVVAVVTACTTT-3' ICAM-1 reverse primer 5'-AGCTTGCACGACCCTTCTAA-3' E-selectin forward primer 5'-CTCCAGGTGAACCAAACAACA-3' E-selectin reverse primer 5'-TGACAACTGCAGGATGCATT-3' ApoB forward primer 5'-ACTGCAGGTCTGGCTCAGGA-3' ApoB reverse primer 5'-CAGGTGGCCACAGCCAATAA-3' MCP1 forward primer 5'-GCATCCACGTGTTGGCTCAG-3' MCP1 reverse primer 5'-TGGACCCATTCCTTCTTGGG-3'

The results of confirming the expression changes of the cell adhesion material, cytokines, and apolipoprotein are shown in Figs. 8a to 8e.

As can be seen in Figures 8a to 8e, the extract of baekjaeja showed superior inhibition of the expression of cell adhesion substances (VCAM-1, ICAM-1, E-selectin) compared to the control. In addition, the expression of cytokines (MCP1) and apolipoprotein (ApoB) was also suppressed. That is, the extract of baekjaeja inhibited the expression of vascular cell adhesion substances and cytokines, which are the causes of arteriosclerosis, and also suppressed the expression of ApoB, which is known as a major risk factor for arteriosclerosis.

In MOVAS cells, the expression of VCAM-1 induced by TNF-a and NF-κB, which is an upper factor, of the extract of quincea extract was measured by western blot.

The results of confirming the expression changes of VCAM-1 and NF-κB are shown in FIG. 9 .

As confirmed in FIG. 9 , it was shown that the extract of baekjaeja inhibited the expression of VCAM-1 induced by TNF-a, and suppressed the expression of NF-κB, a VCAM-1 upstream regulator.

[Formulation example]

On the other hand, the pharmaceutical composition containing the extract according to the present invention can be prepared in various forms depending on the purpose. The following formulations 1 to 4 exemplify a method for preparing a drug containing the extract of baekjae baeja according to the present invention as an active ingredient, but the present invention is not limited thereto.

Formulation 1: Tablet (Direct Pressurization)

After 5.0 mg of the active ingredient was sieved, lactose 14.1 mg, crospovidone USNF 0.8 mg, and magnesium stearate 0.1 mg were mixed and pressurized to make tablets.

Formulation 2: Tablet (wet granulation)

After sieving 5.0 mg of the active ingredient, 16.0 mg of lactose and 4.0 mg of starch were mixed. After dissolving 0.3 mg of polysorbate 80 in pure water, an appropriate amount of this solution was added, followed by atomization. After drying, the fine particles were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The granules were pressurized to make tablets.

Formulation 3: Powders and Capsules

After sieving 5.0 mg of the active ingredient, it was mixed with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. Mix the mixture to a solid No. using a suitable device. Filled in 5 gelatin capsules.

Formulation 4: Injection

An injection was prepared by containing 100 mg as an active ingredient, and also containing 180 mg of mannitol, 26 mg of Na2HPO412H2O, and 2974 mg of distilled water.

In addition, the health food composition containing the extract according to the present invention can be prepared in various forms depending on the purpose. The following formulations 5 to 9 exemplify the manufacturing method of a health food containing the extract of baekjae baeja according to the present invention as an active ingredient, but the present invention is not limited thereto.

Formulation 5. Granular health food

Active ingredient 1000 mg, vitamin A acetate 70 µg, vitamin E 1.0 mg, vitamin 0.13 mg, vitamin B2 0.15 mg, vitamin B6 0.5 mg, vitamin B12 0.2 µg, vitamin C 10 mg, biotin 10 µg, nicotinic acid amide 1.7 mg, folic acid 50 μg, calcium pantothenate 0.5 mg, 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, chloride After mixing 24.8 mg of magnesium, a granular health food was prepared according to a conventional method.

Formulation 6. Health drink

Active ingredient 1000 mg, citric acid 1000 mg, oligosaccharide 100 g, plum concentrate 2 g, and taurine 1 g were mixed, and purified water was added thereto to adjust the total volume to 900 ml. After stirring and heating at 85° C. for about 1 hour, the resulting solution was filtered and obtained in a sterilized 2L container, sealed and sterilized to prepare a health drink.

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.

Formulation 7. Flour Food

0.5 to 5 g of the active ingredient was added to 100 g of wheat flour, and the mixture was used to prepare breads, cakes, cookies, crackers and noodles to prepare health-promoting foods.

Formulation 8. Dairy Products

5 to 10 g of the active ingredient was added to 100 g of milk, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation 9. Seonshik

30 g of brown rice, 20 g of barley, 10 g of glutinous rice, and 15 g of barley radish were pregelatinized by a known method, dried, and roasted, and then prepared as a powder having a particle size of 60 mesh with a grinder. 7 g of black soybeans, 7 g of black sesame, and 7 g of perilla were steamed and dried by a known method, and then dried to prepare a powder having a particle size of 60 mesh with a grinder. A wire meal was prepared by mixing 3 g of the active ingredient of the present invention with the grains and seeds prepared above.

In the above, although the embodiments, experimental examples and formulation examples of the present invention have been described with reference to the accompanying drawings, those of ordinary skill in the art to which the present invention pertains will not change the technical spirit or essential features of the present invention. It will be understood that it may be embodied in specific forms. Therefore, it should be understood that the Examples, Experimental Examples and Formulation Examples described above are illustrative in all respects and not restrictive.

Claims (14)

A composition for preventing, improving or treating obesity comprising an extract of Galla Rhois as an active ingredient.
The method of claim 1,
The active ingredient is adipose tissue weight reduction effect, obesity prevention, improvement or treatment composition.
In the composition for the prevention, improvement or treatment of cardiovascular disease comprising an extract of Galla Rhois as an active ingredient,
The cardiovascular disease is any one or more selected from the group consisting of hypercholesterolemia, hyperlipidemia and arteriosclerosis, a composition for preventing, improving or treating cardiovascular disease.
4. The method of claim 3,
The active ingredient is a composition for the prevention, improvement or treatment of cardiovascular disease, which has an effect of reducing blood cholesterol.
4. The method of claim 3,
The active ingredient inhibits the expression of TNF-a-induced cell adhesion material and apolipoprotein in vascular smooth muscle cells, a composition for preventing, improving or treating cardiovascular disease.
An anti-inflammatory composition comprising an extract of Galla Rhois as an active ingredient.
7. The method according to any one of claims 1 to 6,
The Aphid Aphid ( Melaphis chinensis Bell ) on the leaves of the red tree ( Rhus javanica L. ) or Sumacaceae ( Anacardiaceae ) is a worm nest formed by the stab wound, composition.
7. The method according to any one of claims 1 to 6,
The extract is an extract extracted using water, C ₁ -C ₅ alcohol, or an aqueous solution of C ₁ -C ₅ alcohol, composition.
9. The method of claim 8,
The concentration of the C ₁ -C ₅ alcohol aqueous solution is 45% to 95% (v / v), the composition.
10. The method of claim 9,
The extract is an extract of 75% to 85% (v / v) of an aqueous ethanol solution, composition.
7. The method according to any one of claims 1 to 6,
The composition is a health functional food, composition.
7. The method according to any one of claims 1 to 6,
The composition is a pharmaceutical composition.
13. The method of claim 12,
The pharmaceutical composition is formulated in the form of any one of injections, powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and external preparations, compositions.
7. The method according to any one of claims 1 to 6,
The composition is a pharmaceutical cosmetic composition, composition.

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