KR20200137193A - A composition for improving, preventing and treating obesity and metabolic disease comprising polysaccharide fraction isolated from barley leaf - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating, and treating obesity and metabolic diseases, which can alleviate, prevent or treat diseases caused by metabolic diseases by containing a polysaccharide fraction derived from barley leaves as an active component. In addition, the composition has no toxicity, thereby being able to be consumed in a form of food.

Description

A composition for improving, preventing and treating obesity and metabolic disease comprising polysaccharide fraction isolated from barley leaf}

The present invention relates to a composition capable of improving, preventing or treating obesity and metabolic diseases by containing a polysaccharide fraction derived from barley leaf as an active ingredient.

In modern society, as a result of the change to a meat-oriented diet, the intake of calories has become excessive, while the amount of exercise has become insufficient, and the onset of metabolic diseases including various diseases such as obesity, diabetes, hyperlipidemia, non-alcoholic fatty liver, and dyslipidemia is rapidly increasing.

The obesity refers to a state in which excess energy in the body is excessively accumulated as fat due to a large amount of energy intake compared to the amount of energy consumed. The pharmacological mechanisms of obesity drugs currently used are largely 1) inhibition of fat absorption, 2) promotion of fat breakdown and heat generation, 3) regulation of appetite and satiety, 4) inhibition of protein metabolism, and 5) 　regulation of emotions related to food intake. I can share. Representative obesity treatments include Xenical™, which inhibits fat absorption, and Reductil™, which suppresses appetite by stimulating the sympathetic nervous system. However, in the case of Xenical™, side effects such as fat stool, abdominal pain, vomiting, itching, and liver damage have been reported. In the case of Reductil™, it is believed to cause serious cardiovascular side effects as well as side effects such as headache, loss of appetite, insomnia, and constipation. Have been reported.

Diabetes is a disease that occurs when the secretion of insulin is insufficient or a normal function is not achieved, and is characterized by hyperglycemia in which the concentration of glucose in the blood is high and glucose is excreted through urine. Diabetes treatments currently used include PPAR-γ activators, GLP-1 derivatives, and DPP-IV inhibitors, but these conventional drugs are reported to have side effects showing toxicity to weight gain and liver, kidney, muscle, and heart. have.

The hyperlipidemia is known as a disease that causes inflammation by accumulating on the walls of blood vessels while more fatty substances are present in the blood than necessary, resulting in cardiovascular diseases such as myocardial infarction, stroke or cerebral infarction. Currently used drugs for hyperlipidemia include'statin'-based drugs that have HMG-CoA reductase inhibitory activity, but when used for a long time, they are reported to have side effects showing toxicity to the liver or muscles.

The dyslipidemia refers to a condition in which total cholesterol, LDL cholesterol, or triglyceride in the blood is increased, or a condition in which HDL cholesterol is decreased, and is known to increase the risk of cardiovascular diseases such as heart attack and stroke. Currently, drugs that can be used as treatments for dyslipidemia include Bayer's BAY13-9952 (implitapide), a drug that inhibits the activity of MTP (microsomal triglyceride transfer protein), but these drugs have specific mechanisms of action in cells. There is a problem that it may cause side effects because it is not identified as.

Currently, the development of excellent therapeutic agents for overall obesity or metabolic diseases is inadequate, and various side effects such as the above are also reported for treatments for individual specific diseases.Therefore, the treatment efficacy for obesity or metabolic diseases is excellent, but safe without side effects. There is an urgent need for the development of therapeutic agents.

Korean Patent Registration No. 1,757,385 Korean Patent Registration No. 1,897,770

Non-Patent Document 1. LIU, Gan, et al. Regulation of plasma lipid homeostasis by hepatic lipoprotein lipase in adult mice. Journal of lipid research, 2016, 57.7: 1155-1161.

An object of the present invention is to provide a pharmaceutical composition capable of preventing or treating obesity and metabolic diseases.

In addition, another object of the present invention is to provide a food composition capable of improving or preventing obesity and metabolic diseases.

In order to achieve the above object, the pharmaceutical composition for preventing or treating obesity and metabolic diseases of the present invention may contain a polysaccharide fraction derived from barley leaves as an active ingredient.

The barley leaf-derived polysaccharide fraction contains 65 to 85% by weight of neutral sugar, 5 to 30% by weight of uronic acid, and 2-keto-3-deoxy-D-manno (KDO) relative to the total polysaccharide fraction. -octulosonic acid) KDO-like material may contain 0.5 to 10% by weight.

The polysaccharide fraction derived from barley leaves may further contain 0.5 to 20% by weight of protein.

The uronic acid is composed of galacturonic acid and glucuronic acid; The neutral polysaccharide may include rhamnose, fucose, arabinose, xylose, mannose, galactose, and glucose.

The polysaccharide fraction comprises the steps of (a) treating barley leaf powder with pectin hydrolase; And (b) recovering the fraction by precipitating the enzyme-treated barley leaf powder with C1 to C4 alcohol.

The manufacturing method may further include (c) recovering a fraction having a molecular weight of 50 kDa or more from the polysaccharide fraction of step (b).

The metabolic disease may be one or more selected from the group consisting of dyslipidemia, fatty liver, insulin resistance syndrome, and diabetes, and the dyslipidemia may be one or more selected from the group consisting of hyperlipidemia, hypercholesterolemia, and hypertriglyceridemia. have.

In addition, the food composition for improving or preventing obesity and metabolic diseases of the present invention for achieving the other objects described above may contain a polysaccharide fraction derived from barley leaves as an active ingredient.

The composition for preventing, improving, and treating obesity and metabolic diseases of the present invention reduces body weight, and significantly reduces the expression of factors involved in blood sugar levels and blood triglyceride concentrations that cause metabolic diseases to prevent metabolic diseases. And can be effectively used for treatment.

In addition, the composition of the present invention is non-toxic and can be consumed in the form of food.

1 is a graph showing the results of measuring the average weight of mice divided into groups 1 to 5 for 5 weeks.
2 is a graph showing the results of measuring the average dietary intake of mice divided into groups 1 to 5 for 5 weeks.
3 is a graph showing the results of measuring the weight of the liver of mice divided into groups 1 to 5;
4A is a graph showing the results of measuring the blood glucose concentration of mice divided into groups 1 to 5;
4B is a graph showing the results of measuring blood triglyceride concentrations of mice divided into groups 1 to 5;
FIG. 5 is a graph showing the results of performing an oral fat load test of mice divided into groups 1 to 4 and measuring blood triglyceride concentrations measured for each time period.
6 is a graph showing the calculation of the area under the curve of FIG. 5 (AUC).
7 is a graph measuring the expression level of LPL mRNA in liver tissue of mice divided into groups 1 to 4;

The present invention relates to a composition capable of improving, preventing or treating obesity or metabolic diseases by containing a polysaccharide fraction derived from barley leaf as an active ingredient.

Hereinafter, the present invention will be described in detail.

The polysaccharide fraction derived from barley leaf of the present invention contains 65 to 85% by weight of neutral sugar, 5 to 30% by weight of uronic acid, and 2-keto-3-deoxy-D (KDO) relative to the total polysaccharide fraction. -manno-octulosonic acid) contains 0.5 to 10% by weight of a KDO-like substance. In addition, it may further contain 0.5 to 20% by weight of protein.

The uronic acid is composed of galacturonic acid and glucuronic acid; The neutral polysaccharide may include rhamnose, fucose, arabinose, xylose, mannose, galactose and glucose, but is not limited thereto.

The polysaccharide fraction derived from barley leaf of the present invention comprises the steps of: (a) treating barley leaf powder with pectin hydrolase; And (b) recovering the fraction by precipitating the enzyme-treated barley leaf powder with C1 to C4 alcohol.

First, in step (a), barley leaf powder is treated with pectin hydrolase. The pectin hydrolase may be added in an amount of 1 to 4% by weight, preferably 1 to 3% by weight, and more preferably 1.5 to 2.5% by weight based on the weight of the barley leaf powder.

In the present invention, barley leaves may be undried or dried, and pulverized or powdered may be used.

Preferably, the barley leaf powder treated with the enzyme may be used by suspending it in distilled water at 5 to 20 times (w/v).

The hydrolytic enzyme treatment is preferably performed for 1 to 4 days, more preferably 2 to 3 days.

The step (a) may further perform a process of inactivating the enzyme by heating the remaining pectin hydrolase at 90 to 110° C. for 10 to 60 minutes after the enzyme treatment. Due to the heating, the elution of the soluble polysaccharide component is increased, and the polysaccharide extract obtained by centrifugation and purity is improved by denaturing and precipitating a high molecular protein contained as some impurities.

Since the barley leaf extract obtained through the step (a) undergoes a process of hydrolytic enzyme treatment, the ratio of neutral polysaccharide is remarkably higher than that of the extract obtained by hot water extraction of barley leaf as it is. That is, the barley leaf extract extracted according to the above method contains a large amount of a polysaccharide fraction.

In addition, after step (a), a step of removing the residue from the hydrolyzed barley leaf extract may be additionally performed. The method of removing the residue may be performed by a method of removing solids from a known mixture such as centrifugation and filtration, preferably centrifugation.

Next, in step (b), an organic solvent is added to the enzyme-treated barley leaf hydrolyzate to obtain a precipitated polysaccharide. The organic solvent may be a C1 to C4 alcohol. Alcohol precipitation may be performed by a known ethanol precipitation method, and ethanol used for ethanol precipitation is preferably ethanol having a concentration of 70% to 95% (v/v) by mixing with water.

The manufacturing method may further include (c) recovering a fraction having a molecular weight of 50 kDa or more using ultrafiltration or gel filtration chromatography after step (b).

The method of obtaining the fraction is not particularly limited as long as it is a method of purifying on the basis of molecular weight, but is preferably ultrafiltration or gel filtration chromatography, more preferably gel filtration chromatography. In addition, the form of the final polysaccharide fraction may be an extract, a concentrate, or a powder.

As a result of analyzing the constituent sugar of the obtained barley leaf-derived crude polysaccharide fraction (BLE-0) by gas chromatography, compared to the total polysaccharide, neutral polysaccharide 74.77 wt%, uronic acid 19.37 wt%, protein 3.66 wt%, KDO-like substance 2.19 % By weight; As a result of analyzing the components of the neutral polysaccharide, relative to the total neutral polysaccharide, rhamnose 3.84 mole%, fucose 0.60 mole%, arabinose 16.56 mole%, xylose 37.27 mole%, mannose 1.15 mole%, galactose 11.10 mole%, glucose It was confirmed that the sum of 4.25 mole% and galacturonic acid and glucuronic acid was 19.37 mole%. KDO analogs refer to 2-keto-3-deoxy-D-manno-octulosonic acid (refer to [Table 2]).

A step of purifying the polysaccharide by removing low molecular weight substances and impurities by adding 50 to 100% alcohol having 1 to 4 carbon atoms to the final polysaccharide fraction may be further performed.

Specifically, in the present invention, a barley leaf-derived fraction obtained by treating with the pectinase and precipitating an alcohol having 1 to 4 carbon atoms is obtained by using the same method as described above, and a crude polysaccharide fraction derived from barley leaf having a molecular weight of 50 kDa or more (BLE -0) was obtained.

The metabolic disease may be one or more selected from the group consisting of dyslipidemia, fatty liver, insulin resistance syndrome and diabetes, and the dyslipidemia may be hyperlipidemia, hypercholesteronemia, and hypertriglyceridemia. ) It may be one or more selected from the group consisting of.

The term “fraction” used in referring to barley leaves in the present specification includes not only the fraction obtained by treatment with an extraction solvent, but also the processed product of the polysaccharide fraction derived from barley leaves. For example, the polysaccharide fraction derived from barley leaves may be prepared in a powder state by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

In addition, the barley leaf-derived polysaccharide fraction of the present invention has a meaning to include a barley leaf polysaccharide processed product, for example, barley leaf polysaccharide powder, formulated so that barley leaf can be administered to animals. Although the experiment was conducted with the polysaccharide fraction derived from barley leaf in the present invention, it will be expected by those skilled in the art that the desired effect can be achieved even in the same form as the processed barley leaf polysaccharide.

Meanwhile, in the present specification, the term “contained as an active ingredient” means containing an amount sufficient to achieve the efficacy or activity of the polysaccharide fraction derived from barley leaf. For example, the barley leaf-derived polysaccharide fraction is used in a concentration of 10 to 1500 μg/ml, preferably 100 to 1000 μg/ml. Since the barley leaf-derived polysaccharide fraction is a natural product and does not have side effects on the human body even if it is administered in excess, the upper limit of the quantity of the barley leaf-derived polysaccharide fraction included in the composition of the present invention can be selected and carried out within an appropriate range.

The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, It is possible to use a lubricant or flavoring agent.

For administration, the pharmaceutical composition may contain one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.

The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, sterilization and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

Furthermore, it can be preferably formulated according to each disease or ingredient using a method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration. .

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, Usually, the skilled practitioner can readily determine and prescribe the dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention may be prepared in a unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient, or may be prepared by placing it in a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

In addition, the present invention provides a food composition for improving or preventing obesity or metabolic diseases containing a polysaccharide fraction derived from barley leaves as an active ingredient.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, health supplements. Etc.

The food composition of the present invention may include not only a barley leaf-derived polysaccharide fraction as an active ingredient, but also an ingredient commonly added during food production, for example, protein, carbohydrate, fat, nutrients, seasoning and flavoring agents. Include. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)] and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts, etc., in addition to the polysaccharide fraction derived from barley leaf of the present invention may be additionally included. I can.

The present invention provides a health functional food comprising a food composition for improving, preventing or treating obesity or metabolic diseases comprising the barley leaf-derived polysaccharide fraction as an active ingredient. Health functional foods are foods prepared by adding a polysaccharide fraction derived from barley leaf to food materials such as beverages, teas, spices, gums, confectionery, or encapsulated, powdered, or suspended. It means to bring it, but unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long time by using food as raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of polysaccharide fraction derived from barley leaf in such health functional foods cannot be uniformly regulated depending on the type of health functional food to be targeted, but can be added within the range that does not damage the original taste of the food. It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In addition, the present invention provides a use of a polysaccharide fraction derived from barley leaf for the manufacture of a pharmaceutical or food for improvement, prevention or treatment of obesity or metabolic disease. As described above, the barley leaf-derived polysaccharide fraction may be used for improvement, prevention or treatment of obesity or metabolic diseases.

In addition, the present invention provides a method for improving, preventing or treating obesity or metabolic diseases, comprising administering an effective amount of a polysaccharide fraction derived from barley leaves to a mammal.

The term "mammal" as used herein refers to a mammal that is the subject of treatment, observation or experimentation, and preferably refers to a human.

The term "effective amount" as used herein refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, which is considered by a researcher, veterinarian, doctor or other clinician, Includes an amount that induces relief of symptoms of the disease or disorder. The effective amount and the number of administrations for the active ingredient of the present invention may vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the amount of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient. It can be adjusted according to various factors including condition, sex and diet, time of administration, route of administration and secretion rate of the composition, duration of treatment, and drugs used simultaneously. In the prophylaxis, treatment or improvement method of the present invention, in the case of adults, it is preferable to administer the polysaccharide fraction derived from barley leaves at a dose of 0.001 g/kg to 10 g/kg when administered once to several times a day.

In the treatment method of the present invention, the composition comprising a polysaccharide fraction derived from barley leaves as an active ingredient is conventionally used through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a phosphorus manner.

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention, but the following examples are only illustrative of the present invention, and it is obvious to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It is natural that such modifications and modifications fall within the appended claims.

Example 1. Barley leaf-derived crude polysaccharide fraction (BLE-0)

Barley grass was produced in Yeonggwang, Jeollanam-do, and was purchased and used by Saemomwon Co., Ltd.

Powdered barley was suspended in distilled water 10 times (w/v) (pH 4), and pectinase (Rapidase C80MAX, Vision Biochem) was added at 1% per raw material, followed by enzymatic treatment in a 50° C. incubator for 3 days. Thereafter, the enzyme-treated reaction solution was heated at 100° C. for 15 minutes to inactivate the remaining pectinase.

After the enzyme treatment was completed, the sample was centrifuged at 4° C., 6,500×g for 15 minutes to remove the residue, and 80% ethanol was added thereto to allow the polysaccharide to precipitate while standing for 4 hours. After recovering the precipitated polysaccharide, the precipitated polysaccharide was dissolved in a small amount of distilled water, and then low-molecular substances were removed using a dialysis membrane (MWCO 3,500), and an enzyme-treated polysaccharide fraction (BLE-0) was obtained.

Comparative Example 1. Crude polysaccharide fraction derived from barley leaves (BLW-0)_non-enzyme treatment (hot water extraction)

Powdered barley was suspended in distilled water 10 times (w/v) and heated at 100° C. for 3 hours. After centrifugation of the hot water extract at 4° C., 6,500×g, 15 minutes, the supernatant was separated, 80% ethanol of 4 times the volume (v/v) of the supernatant was added, and the polysaccharide was precipitated while standing for 4 hours. Dialysis (MWCO 6,000-8,000) was performed on the precipitated polysaccharide to obtain a crude polysaccharide fraction (BLW-0) extracted with hot water.

<Test Example>

Test Example 1. Analysis per composition of fractions

Constituent sugar analysis is performed by partially modifying the method of Albersheim et al. to derivatize each constituent sugar with alditol acetate and aldonolactone after hydrolysis, and then GC (Gas Chromatography ACME-6100, Young-Lin Co. Ltd. Anyang, Korea). ) Was used. The polysaccharide sample was hydrolyzed by reacting for 1.5 hr at 121° C. in 2 M TFA (trifluroacetic acid), dissolved in 1 ml of 1 M NH ₄ OH (ammonia solution), and reduced with 10 mg of NaBH ₄ for 4 hr. After adding an appropriate amount of acetic acid to remove the remaining NaBH ₄ , the mixture was repeatedly dried with methanol to remove excess acetic acid and converted to alditol corresponding to each constituent sugar. To find out the composition of neutral sugar, 1 ml of acetic anhydride was added to each alditol and reacted at 121° C. for 30 min to convert to alditol acetate, which was converted into chloroform/H ₂ O It was separated and extracted with a two-phase solvent system, and the extract was dried and dissolved in a small amount of acetone to be used as a sample for GC analysis.

In addition, in order to analyze the composition of acidic sugars, the converted alditol was dissolved in distilled water, developed on a Sep-pak QMA Cartridge (Waters, Milford, MA, USA), and aldonic acid was separated with 1 M HCl. 2 M TFA was added to the separated solution and reacted at 100° C. for 5 min to convert to aldonolactone. After that, it was separated and extracted with a chloroform/H ₂ O two-phase solvent system, dried and used as a GC analysis sample, and the GC column was an SP-2380 capillary column (0.25 ㎜×30 m, 0.2 ㎛ film thickness, Supelco, Bellefonte, PA , USA), a flame ionization detector (FID, Young-Lin Co. Ltd.) was used as the detector, and N ₂ as a carrier gas was flowed at 1.5 ml/min. Injection temperature of 250 ℃, detector temperature of 270 ℃, column temperature from 60 ℃ to 220 ℃ 30 ℃ / min, 220 ℃ to 250 ℃ under the conditions of 8 ℃ / min (see [Table 1]). The mole% of each component was calculated from the peak area ratio of each derivative, the reaction coefficient for the flame ionization detector (FID), and the molecular weight of the alditol acetate derivative of each component.

Apparatus GC ACME-6100 (Young-Lin Co. Ltd., Anyang, Korea) Detector Flame ionization detector(FID) (Young-Lin Co. Ltd., Anyang, Korea) Column SP-2380 capillary column (Supelco, Bellefonte, USA) Column size 0.25 mm × 30 m, 0.2 m film thickness Oven temp. 60 ℃(1min) → 220 ℃(12min) → 250 ℃(15min) 30 ℃/min 8 ℃/min Injector temp. 250 ℃ Detector temp. 270 ℃ Carrier gas N ₂ (1.5 mL/min)

As a result, as a result of analyzing the physicochemical properties such as the main component content and composition sugar composition of the hot water extract polysaccharide fraction (BLW-O) and RAPIDASE C80MAX (Pectinase) enzyme-treated polysaccharide fraction (BLE-0) from barley leaves, the following table It was like 2.

Chemical composition (unit: wt%) Example 1 (BLE-0) Comparative Example 1 (BLW-0) Neutral sugar 74.77 ± 5.07 42.22 ± 4.60 Uronic acid 19.37 ± 0.47 44.64 ± 2.27 Protein 3.66 ± 0.53 11.54 ± 0.76 KDO-liked material 2.19 ± 0.22 1.60 ± 0.11 Sugar composition (unit: mole%) Rhamnose 3.84 ± 0.04 2.60 ± 0.05 Fucose 0.60 ± 0.02 - Arabinose 16.56 ± 0.13 15.71 ± 0.20 Xylose 37.27 ± 0.07 5.39 ± 0.06 Mannose 1.15 ± 0.05 - Galactose 11.10 ± 0.11 14.78 ± 0.09 Glucose 4.25 ± 0.07 3.75 ± 0.13 GalA+GlcA 19.37 ± 0.47 44.64 ± 2.27

* In the above table, KDO-linked material (KDO-like substance) means 2-keto-3-deoxy-D-manno-octulosonic acid, and “GalA+GlcA” means the sum of galacturonic acid and glucuronic acid. Make up Ronic acid.

* In the table above, the sugar composition was analyzed using the alditol acetates derivative method.

* The content ratio (%) of the chemical component represents the ratio in the dry sample.

* The mole% is calculated from the amount of total neutral sugar detected.

As a result of analysis of general components and constituents of the polysaccharide fractions, Comparative Example 1 (BLW-0), which is a non-enzyme-treated polymeric polysaccharide fraction, contains an acidic sugar (42.22%) and an acidic sugar (44.64%) at almost 1:1. Although shown as a polysaccharide fraction, Example 1 (BLE-0), which is an enzyme-treated polymeric polysaccharide fraction, was analyzed as a neutral polysaccharide fraction in which the content of protein (3.66%) was rapidly reduced together with acidic sugar (19.37%) (Table). 2).

This is due to the enzymatic action of the commercially available Rapidase ^TM , where pectinase is the main activity, and polygalacturonan [(α-D-galacturonic acid)n], which mainly comprises homogalacturonan (HG), was decomposed and removed by dialysis. In addition, in the composition of constituent sugars, Example 1 (BLE-0) mainly contains xylose, arabinose, and galactose (36.09, 16.04 and 10.74%) as well as a small amount of rhamnose. In addition to the arabinogalactan constituting ), the possibility of the existence of arabinoxylan, which is classified as hemicellulose, could be strongly deduced.

Test Example 2. Animal experiment

Experimental animals

22g of 6-week-old C57BL/6J male mice (Joongang Animal Lab, Seoul, Korea) were used in the experiment, and the mice were subjected to acclimatization for 2 weeks before the experiment. Mice were reared on a free diet in a breeding room controlled at 22±1°C temperature and 55%±10% humidity throughout the experiment, and the light/dark cycle was controlled at a 12 hour cycle. All animal experiments were conducted in accordance with the Animal Experiment Operation Regulations of Korea Food Research Institute.

Sample administration and sampling

For administration of the sample, the mice were randomly divided into five groups. ND (normal diet) was a negative control group that ate a normal diet, and the HFD and sample administration groups were allowed to autonomously eat a high fat diet (TD.88137, Teklad Laboratories, Madison, WI, USA). Samples were administered orally with 200 mg/kg of BLE0 and 10 g/kg of fish oil as a positive control, and 200 mg/kg of sample BLW0 was administered, and the same amount of distilled water was used in the ND and HFD groups for the same period. It was administered orally during the period. Weight (FIG. 1) and dietary intake (FIG. 2) were measured every week for a total of 5 weeks, and at the end of the experiment, the liver was removed under anesthesia and the weight was measured (FIG. 3). For statistical analysis of the experiment, a one-way ANOVA (Dunnett's test vs. HFD) was used to determine the significant difference in mean values between groups, and a p value lower than 0.05 was considered as a significant result. To indicate the statistical significance between each group and HFD, the significant difference between ND and HFD is indicated as #, the difference between BLE0 and HFD is indicated as *, and the difference from fish oil is indicated as §.

-5 groups of mice-

Group 1 (ND, negative control): normal diet intake, oral administration of distilled water

Group 2 (HFD): high fat diet intake, oral administration of distilled water

Group 3 (BLE0): high fat diet intake, enzyme-treated crude polysaccharide fraction (BLE-0) 0.20 g/kg orally administered

Group 4 (Fish oil): high fat diet intake, figh oil 10 g/kg orally administered

Group 5 (BLW-0): High fat diet intake, hot water extracted crude polysaccharide fraction (BLW-0) 0.20 g/kg orally administered

Test Example 2-1. Animal model weight measurement

1 is a graph showing the results of measuring the average weight of mice in groups 1 to 5 for 5 weeks.

As shown in FIG. 1, significant weight reduction efficacy under high fat diet intake in the experimental group in which 200 mg/kg of the enzyme-treated crude polysaccharide fraction (BLE0) prepared according to Example 1 of the present invention was orally administered daily for 5 weeks This is showing that there is.

Compared to the HFD group that consumed only the high-fat diet, the ND group that ate a normal diet showed a lower average weight from 1 week after dietary change, and when administered BLE0 and fish oil, the average weight decreased significantly compared to the HFD group from 2 weeks later. The results were confirmed.

In addition, in the case of the experimental group administered with the hot water extracted crude polysaccharide fraction (BLW0), a significant weight reduction could not be confirmed.

Test Example 2-2. Dietary intake measurement

2 is a graph showing the results of confirming the average dietary intake of mice in groups 1 to 5 for 5 weeks. As shown in FIG. 2, the dietary intake in the HFD group increased by about 38% on a calorie basis compared to the ND group, and when fish oil was administered, the dietary intake decreased by 19.4% compared to HFD. However, when administering the enzyme-treated crude polysaccharide fraction (BLE0) prepared according to Example 1 of the present invention, there was no significant difference, and it can be seen that weight reduction compared to HFD was significantly observed without reduction in dietary intake. On the other hand, even when the hot water extracted crude polysaccharide fraction (BLW0) prepared according to Comparative Example 1 of the present invention was administered, a significant difference in dietary intake could not be confirmed.

Test Example 2-3. Liver weight measurement

3 is a graph showing the results of measuring the weight of the liver by extracting the liver of the mice in groups 1 to 5; It is known that the increase in liver weight caused by the intake of a high fat diet is due to fatty liver, such as the production of fat inside the liver tissue, and in fact, the liver weight of the HFD group shows a significant increase compared to the ND group. The BLE0 and fish oil-administered groups showed similar liver weights compared to the ND group, and were significantly different from the HFD group. On the other hand, in the case of the group to which the hot water-extracted crude polysaccharide fraction (BLW0) prepared according to Comparative Example 1 of the present invention was administered, the liver weight increased compared to the ND group, and there was no significant difference compared to the HFD group.

Test Example 2-4. Improve blood sugar and blood fat metabolism

1) Analysis of blood index

In order to confirm the efficacy of improving dyslipidemia and related metabolic diseases by administration of an enzyme-treated crude polysaccharide fraction (BLE0) prepared according to Example 1 of the present invention, a blood sample obtained from the caudal vein of a mouse maintained fasting for 15 hours or longer was obtained. Used. For blood glucose measurement, a trace amount (2 μl) of whole blood was collected from the caudal vein of the experimental animal, and then blood glucose was measured and recorded using a blood glucose meter (Accu-Chek Performa, Roche Diagnostics, Indianapolis, IN, USA). Blood was collected in a heparin-treated tube, centrifuged at 10,000 rpm for 10 minutes to separate the serum, and stored at -80°C until used in an experiment for analysis of triglyceride (TG) in blood. To measure blood triglycerides, a serum triglyceride determination kit (Sigma-Aldrich, St. Louis, MO, USA) was used to perform the manufacturer's protocol. For statistical analysis of the experiment, a one-way ANOVA (Dunnett's test vs. HFD) was used to determine the significant difference in mean values between groups, and a p value lower than 0.05 was considered as a significant result. To indicate the statistical significance between each group and HFD, the significant difference between ND and HFD is indicated as #, the difference between BLE0 and HFD is indicated as *, and the difference from fish oil is indicated as §.

4A is a graph showing plasma glucose concentrations of mice divided into groups 1 to 5, and FIG. 4B is a graph showing blood triglyceride concentrations of mice divided into groups 1 to 5.

As shown in FIGS. 4A and 4B, it can be seen that both blood sugar and triglycerides were significantly increased in the HFD group that ate a high-fat diet compared to ND that consumed a normal diet, and BLE0 200 mg/kg was administered. It can be seen that this increase decreases significantly. It was also confirmed that the blood triglyceride was also decreased when the positive control fish oil was administered. On the other hand, in the group administered with BLW0, neither blood sugar nor blood triglycerides significantly decreased.

2) Oral olive oil tolerance test

In order to more accurately check the effect of improving blood triglyceride metabolism by administering a sample, an oral fat load test that checks whether triglyceride metabolism is properly performed by measuring triglyceride in blood after forcibly administering triglyceride (olive oil). tolerance test) was performed. For this, the mice were kept fasting for more than 15 hours, and blood was collected from the caudal vein after 0, 1.5, 3, and 6 hours after oral administration of 5 ml/kg of olive oil. The collected blood was centrifuged (12,000 rpm, 30 seconds) and the supernatant was stored at -80 °C. Blood triglycerides were measured according to the manufacturer's protocol using a serum triglyceride determination kit (Sigma-Aldrich, St. Louis, MO, USA). The blood triglyceride concentration measured by time is shown in Fig. 5 as an average value and a standard error for each group, and the area under curver (AUC) is calculated in units of mg/dl*hr and is shown as a graph ( Fig. 6). For statistical analysis of the experiment, a one-way ANOVA (Dunnett's test vs. HFD) was used to determine the significant difference in mean values between groups, and a p value lower than 0.05 was considered as a significant result. To indicate the statistical significance between each group and HFD, the significant difference between ND and HFD is indicated as #, the difference between BLE0 and HFD is indicated as *, and the difference from fish oil is indicated as §.

The BLW0 group was judged to have no effect on dyslipidemia and was not used in subsequent experiments.

FIG. 5 is a graph obtained by measuring blood triglyceride concentrations of mice divided into groups 1 to 4 by time, and FIG. 5 is a graph calculating an area under the curve (AUC) of the graph of FIG. 4.

As shown in FIGS. 5 and 6, it was confirmed that blood triglyceride concentration was higher in the HFD group compared to ND, and this increase was decreased when BLE0 administration. However, since fish oil, which was a positive control, contained oil as a sample itself, this olive oil tolerance test, which evaluates triglyceride metabolism in a short period of time, did not show any effect.

As a result of this experiment, it was confirmed that BLE0 has an effect of lowering triglycerides in the normal blood, and in particular, has the effect of clearing it when an excessive amount of olive oil is administered for a short period of time.

Test Example 2-5. Analysis of regulatory mechanisms related to lipid metabolism

In order to more accurately confirm the effect of improving blood triglyceride metabolism by sample administration, the relevant mechanism was explored. Lipoprotein lipase (LPL) was selected as an important factor that can control the concentration of triglyceride in blood. In relation to the expression of LPL in the liver, it is known through prior studies that the expression of LPL in the liver directly regulates the blood triglyceride concentration after a meal.

To confirm this, immediately after sacrifice of the mouse, the liver tissue was immersed in liquid nitrogen and stored at -80 °C. 300 mg of each liver tissue was used, RNA was extracted using Qiagen RNA purification kit (Life Technologies, Inc.), and cDNA was synthesized using iScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA). . In order to measure the relative expression level of LPL mRNA in tissues, ABI Step One Plus Real-Time PCR System (Life Technologies, Inc.) was used as a special primer pair for LPL detection (for.word: 5'-TTGCCCTAAGGACCCCTGAA-3', reverse: 5'-ACAGAGTCTGCTAATCCAGGAAT-3') was used. For data analysis and processing, software provided by PCR company (StepOne Software, v2.3, Life Technologies) was used. For statistical analysis of the experiment, a one-way ANOVA (Dunnett's test vs. HFD) was used to determine the significant difference in mean values between groups, and a p value lower than 0.05 was considered as a significant result. To indicate the statistical significance between each group and HFD, the significant difference between ND and HFD is indicated by #, and the difference between BLE0 and HFD is indicated by *.

7 is a graph measuring the expression level of LPL mRNA in liver tissues of mice divided into groups 1 to 4;

As shown in Figure 7, it can be seen that the expression of LPL in the liver compared to HFD is significantly lower in the case of a normal diet (ND), and it is confirmed that this is restored to a normal value compared to the HFD group when administered with BLE0. I did.

Test Example 3. Return mutation toxicity test

In order to confirm the toxicity by the reversion mutation test of the crude polysaccharide fraction (BLE-0) prepared according to Example 1, the method suggested by Maron and Ames (1983) described in the literature was partially modified and tested as follows ( Maron, DM and Ames BN (1983) Revised methods for the Salmonella mutagenicity test, Mutat. Res. 113: 173-215).

To search for mutagenicity for specific components, strains (Salmonella typhimurium) TA98, TA100, TA1535, and TA1537 were used. Treatment of the test substance was performed by a direct plate incorporation method in which metabolic active enzyme system (S-9 mix) was applied or not applied. Salmonella typhimurium TA98 strain used in the reversion mutation test was purchased from Molecular Toxicology Inc. (USA). For the strain, inoculate 50 μL of the frozen test strain solution into 25 mL of liquid medium (2.5% Oxoid Nutrient broth No. 2), and use a shaking incubator (VS-8480SFN, Vision Science Co., Ltd.). It was used after incubating for about 10 hours at ℃. The minimal glucose agar plate was prepared with 1.5% Bacto agar (214010, BD Difco), Vogel-Bonner medium E, and 2% glucose, and the top agar was prepared with 0.6% agar and 0.5% NaCl. , 0.05 mM histidine (43011, Fluka)-biotin (47868, Supelco) was added to the top agar.

Dispense 2 mL each of the autoclaved top agar into a sterilized tube preheated to 45°C in a dry bath (11-718-4, Fisher Scientific), and 0.1 mL of the test substance solution and bacterial culture solution. 0.1 mL was mixed with a top agar, and immediately shaken with a vortex mixer (37600, Thermolyne) for 2-3 seconds, poured into a minimal glucose agar plate, tilted in various directions, and then solidified. For the excipient group (negative control), 0.1 mL of the excipient was added instead of the test material solution, and the positive control solution (2-Aminoanthracene (2AA), standard: Sigma A1381) was added in the same manner to the positive control group. After the top agar was hardened, the plate was turned over and incubated at 37° C. for about 48 hours with the plate lid closed, and colonies were counted.

Strain Metabolic activity
Enzyme system Return mutant count/plate 0 μg/plate 500 μg/plate 1000 μg/plate TA98 -S9 20±1 23±0 18±4 +S9 24±3 24±1 21±0 TA100 -S9 135±6 140±8 121±6 +S9 145±4 134±5 142±6 TA1535 -S9 12±1 9±1 13±3 +S9 12±1 11±1 12±1 TA1537 -S9 12±1 10±1 11±1 +S9 14±1 9±1 11±2

As shown in Table 3 above, it was confirmed that the crude polysaccharide fraction (BLE-0) prepared according to Example 1 of the present invention did not induce a return mutation for the strain used under this test condition.

Test Example 4. In vitro staining abnormality test using Chinese Hamster Lung (CHL) cells

As an experiment to determine whether the crude polysaccharide fraction (BLE-0) prepared according to Example 1 causes structural or numerical abnormalities in the chromosomes of cultured Chinese Hamster Lung (CHL) cells, Chinese hamster lung (CHL) cells were used. Thus, a chromosomal abnormality test was performed with and without the application of the metabolic activity system. As a metabolic activity system, a cofactor was added to the rat liver homogenous fluid induced by Aroclor-1254.

The test substance was suspended and treated in a physiological saline injection solution. The highest concentration was determined using Relative Increased Cell Count (RICC) as an index of cytotoxicity. The concentration group was set as shown in the following table, including the negative (excipient) control group and the positive control group, and one flask per concentration group was used.

Actively proliferating cells were isolated, 5 x 10 ⁴ cells were seeded in 5 mL of culture medium in a flask with a culture area of 25 cm 2, cultured for about 3 days, and then the test material was treated. Chromosomal samples were prepared 24 hours after the start of treatment, and chromosomal abnormalities were counted from 150 metaphases per concentration group.

6 hours treatment with metabolic activation system

Relative Increased Cell Count (RICC) was calculated by the following equation from the number of cells obtained by separating and counting cells from the flask, and used as an index of cytotoxicity.

[Equation 1]

S9 density
(μg/mL) Cell count RICC(%) Abnormal medium weather
(%) PP+ER(%) + 0 6388±123 100 0.00 0.00 31.25 6224±50 96 0.00 0.00 62.5 6297±139 98 0.00 0.00 125 5932±33 89 0.00 0.00 250 5943±17 89 0.00 0.00 500 6086±11 93 0.00 0.00 1000 5675±68 83 0.00 0.00 Benzo[a]pyrene 4418±246 52 34.00 0.00

Cloudiness was observed in the concentration group of 500 μg/mL or higher of the test substance. As shown in Table 4 above, the frequency of structural abnormal intermediate phase (excluding gap) was negative control, test substance 31.25, 62.5, 125, 250, 500 and 1000. In the order of μg/mL, 0.00, 0.00, 0.00, 0.00, 0.00, 0.00, and 0.00% showed no statistically significant increase in all concentration groups treated with the test substance compared to the negative control group, and there was no dose correlation. The incidence of mid-phase phase with a number of abnormalities was 0.00% in both the negative control group and all test substance treatment groups, and there was no statistically significant increase in all concentration groups treated with the test substance compared to the negative control group, and there was no dose correlation.

In the positive control group, a statistically significant increase was observed in the frequency of metaphase (34.00%) with structural abnormalities (P<0.01).

6 hours treatment without metabolic activation system

S9 density
(μg/mL) Cell count RICC(%) Abnormal medium weather
(%) PP+ER(%) - 0 8694±16 100 0.00 0.00 31.25 7894±55 88 0.00 0.00 62.5 7882±109 87 0.00 0.00 125 8097±7 91 0.00 0.00 250 8043±57 90 0.00 0.00 500 7963±109 89 0.00 0.00 1000 7206±3 77 0.00 0.00 4-Nitroquinoline-1-oxide 0.4 7306±26 78 12.00 0.00

As shown in Table 5 above, the frequency of structural abnormal intermediate phases is 0.00, 0.00, 0.00, 0.00, 0.00, 0.00 and 0.00% in the order of negative control, test substance 31.25, 62.5, 125, 250, 500, and 1000 μg/mL. , There was no statistically significant increase in all concentration groups treated with the test substance compared to the negative control group, and there was no dose correlation. The incidence of mid-phase phase with a number abnormality was 0.00% in both the negative control group and all test substance treatment groups, and there was no statistically significant increase in all concentration groups treated with the test substance compared to the negative control group, and there was no dose correlation.

In the positive control group, a statistically significant increase was observed in the frequency of metaphase (12.00%) with structural abnormalities (P<0.01).

24-hour treatment without metabolic activation system

S9 density
(μg/mL) Cell count RICC(%) Abnormal medium weather
(%) PP+ER(%) - 0 7642±51 100 0.00 0.00 31.25 7758±6 102 0.00 0.00 62.5 7527±3 98 0.00 0.00 125 7598±52 99 0.00 0.00 250 7227±44 92 0.00 0.00 500 6238±29 74 0.00 0.00 1000 5941±50 68 0.00 0.00 4-Nitroquinoline-1-oxide 0.4 6951±30 87 13.33 0.00

As shown in Table 6 above, the frequency of structural abnormal intermediate phases is 0.00, 0.00, 0.00, 0.00, 0.00, 0.00 and 0.00% in the order of negative control, test substance 31.25, 62.5, 125, 250, 500, and 1000 μg/mL. , There was no statistically significant increase in all concentration groups treated with the test substance compared to the negative control group, and there was no dose correlation. The incidence of mid-phase phase with a number abnormality was 0.00% in both the negative control group and all test substance treatment groups, and there was no statistically significant increase in all concentration groups treated with the test substance compared to the negative control group, and there was no dose correlation.

In the positive control group, a statistically significant increase was observed in the frequency of metaphase (13.33%) with structural abnormalities (P<0.01).

As a result of counting chromosomal abnormalities, the appearance frequency of the intermediate phase with chromosomal abnormalities in all test substance treatment groups did not increase compared to the negative control group, regardless of the treatment method, and this result did not satisfy the positive criteria.

Therefore, it was confirmed that the crude polysaccharide fraction (BLE-O), which is a test substance, does not cause chromosomal abnormalities in CHL cells under this test condition.

Hereinafter, examples of the formulation of the composition containing the powder of the present invention will be described, but the present invention is not intended to limit this, but is intended to be described in detail.

Formulation Example 1. Preparation of powder

500 mg of crude polysaccharide fraction derived from barley leaves obtained in Example 1

100 mg lactose

10 mg of talc

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2. Preparation of tablet

300 mg of crude polysaccharide fraction derived from barley leaves obtained in Example 1

100 mg corn starch

100 mg lactose

2 mg of magnesium stearate

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

Formulation Example 3. Preparation of Capsule

200 mg of crude polysaccharide fraction derived from barley leaves obtained in Example 1

3 mg of crystalline cellulose

14.8 mg lactose

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.

Formulation Example 4. Preparation of injection

600 mg of crude polysaccharide fraction derived from barley leaves obtained in Example 1

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Na ₂ HPO _4, 12H ₂ O 26 mg

It is prepared with the above ingredients per ampoule according to a conventional injection preparation method.

Formulation Example 5. Preparation of liquid formulation

4 g of crude polysaccharide fraction derived from barley leaves obtained in Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of liquid preparation, add each ingredient to purified water to dissolve it, add lemon zest, mix the above ingredients, add purified water and add purified water to adjust the total to 100g, then fill in a brown bottle for sterilization. To prepare a solution.

Formulation Example 6. Preparation of granules

1,000 mg of crude polysaccharide fraction derived from barley leaves obtained in Example 1

Vitamin mixture right amount

Vitamin A acetate 70 ㎍

1.0 mg of vitamin E

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

1.7 mg of nicotinic acid amide

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

1.75 mg ferrous sulfate

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

90 mg of potassium citrate

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for granules, but it is possible to arbitrarily modify the mixing ratio. After mixing the above ingredients according to a conventional granule preparation method, granules And can be used to prepare a health functional food composition according to a conventional method.

Formulation Example 7. Preparation of functional beverage

1,000 mg of crude polysaccharide fraction derived from barley leaves obtained in Example 1

1,000 mg citric acid

100 g oligosaccharides

2 g of plum concentrate

1 g taurine

Total 900 mL with purified water

After mixing the above ingredients according to a normal health drink manufacturing method, stirring and heating the resulting solution at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and stored in a refrigerator. It is used to prepare the functional beverage composition of the invention.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences, such as the demand class, the country of demand, and the purpose of use.

Claims (15)

A pharmaceutical composition for preventing or treating obesity or metabolic disease, characterized in that it contains a polysaccharide fraction derived from barley leaf as an active ingredient. The method of claim 1, wherein the barley leaf-derived polysaccharide fraction contains 65 to 85% by weight of neutral sugar, 5 to 30% by weight of uronic acid, and 2-keto-3 (KDO) relative to the total polysaccharide fraction. -deoxy-D-manno-octulosonic acid) KDO-like material comprising 0.5 to 10% by weight of obesity or a pharmaceutical composition for the prevention or treatment of metabolic diseases, characterized in that it contains. The pharmaceutical composition for preventing or treating obesity or metabolic diseases according to claim 2, wherein the barley leaf-derived polysaccharide fraction further contains 0.5 to 20% by weight of protein. The pharmaceutical composition for preventing or treating obesity or metabolic diseases according to claim 2, wherein the uronic acid is composed of galacturonic acid and glucuronic acid. The pharmaceutical composition for preventing or treating obesity or metabolic diseases according to claim 2, wherein the neutral polysaccharide comprises rhamnose, fucose, arabinose, xylose, mannose, galactose, and glucose. The method of claim 1, wherein the polysaccharide fraction comprises the steps of: (a) treating barley leaf powder with pectin hydrolase; And
(b) recovering the fraction by precipitating the enzyme-treated barley leaf powder with C1 to C4 alcohol. A pharmaceutical composition for preventing or treating obesity or metabolic disease, characterized in that prepared by a manufacturing method comprising. The method of claim 6, wherein (c) recovering a fraction having a molecular weight of 50 kDa or more from the polysaccharide fraction of step (b); prevention of obesity or metabolic disease, characterized in that it is prepared by a manufacturing method further comprising Or a pharmaceutical composition for treatment. The pharmaceutical composition for preventing or treating obesity or metabolic disease according to claim 1, wherein the metabolic disease is at least one selected from the group consisting of dyslipidemia, fatty liver, insulin resistance syndrome, and diabetes. The pharmaceutical composition for preventing or treating obesity or metabolic disease according to claim 8, wherein the dyslipidemia is at least one selected from the group consisting of hyperlipidemia, hypercholesterolemia, and hypertriglyceridemia. A food composition for improving or preventing obesity or metabolic diseases, characterized in that it contains a polysaccharide fraction derived from barley leaves as an active ingredient. The method of claim 10, wherein the barley leaf-derived polysaccharide fraction contains 65 to 85% by weight of neutral sugar, 5 to 30% by weight of uronic acid, and 2-keto-3 (KDO) relative to the total polysaccharide fraction. -deoxy-D-manno-octulosonic acid) KDO-like material consisting of 0.5 to 10% by weight of obesity or metabolic disease improvement or prevention food composition, characterized in that it contains. The food composition for improving or preventing obesity or metabolic disease according to claim 11, wherein the barley leaf-derived polysaccharide fraction further comprises 0.5 to 20% by weight of protein. The food composition for improving or preventing obesity or metabolic diseases according to claim 11, wherein the uronic acid is composed of galacturonic acid and glucuronic acid. The food composition for improving or preventing obesity or metabolic disease according to claim 11, wherein the neutral polysaccharide comprises rhamnose, fucose, arabinose, xylose, mannose, galactose, and glucose. The food composition for improving or preventing obesity or metabolic disease according to claim 10, wherein the metabolic disease is at least one selected from the group consisting of dyslipidemia, fatty liver, insulin resistance syndrome, and diabetes.

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