KR102798916B1 - Composition for preventing, treating, or alleviating obesity or obesity-related disease comprising Setaria viridis extracts as an active ingredient - Google Patents

Abstract

The present invention relates to a composition containing a dog's paw extract as an effective ingredient for preventing, treating, or improving obesity or obesity-related complications. It was confirmed that the dog's paw extract according to the present invention has the effect of suppressing the increase in body weight and body fat caused by a high-fat diet in a mouse model induced by dietary obesity, and improving oxidative damage. Furthermore, it was specifically confirmed that it has the effect of improving abnormal lipids and fatty liver, hepatotoxicity, and fibrosis. Therefore, it is expected that the dog's paw extract can be usefully used not only for improving obesity and body fat, but also for preventing, treating, or improving obesity-related complications that may be caused by weight gain.

Description

Composition for preventing, treating, or alleviating obesity or obesity-related disease comprising Setaria viridis extracts as an active ingredient

The present invention relates to a composition for preventing, treating, or improving obesity or obesity-related complications, comprising a dog's-eye extract as an active ingredient.

Obesity generally refers to a state of excessive adipose tissue in the body, and it is a phenomenon in which the energy consumed through food is not balanced with the energy consumed through physical activity, so that the excess energy is stored as body fat. Obesity is caused by a variety of factors, including genetic factors, environmental factors such as irregular eating habits, lack of exercise, or excessive consumption of fast food, psychological factors such as depression, boredom, and excessive stress, and pathological factors such as changes in thyroid or adrenal cortex hormones. Recently, with economic growth and changes in lifestyle, many changes have occurred in eating habits, and busy modern people are experiencing an increase in body weight and obesity due to high-calorie diets such as fast food and little exercise.

Obesity is recognized as more serious because of the various complications that can be caused by obesity rather than the danger itself, and if body fat increases abnormally due to energy imbalance over a long period of time, it can cause various metabolic diseases and adult diseases such as diabetes, hyperlipidemia, heart disease, stroke, arteriosclerosis, and fatty liver. In addition, obesity can cause mental illness such as social isolation or alienation, lack of confidence, and depression, so obesity is emerging as a serious social problem worldwide, and the need for prevention and treatment of obesity is recognized as very important.

Obesity can be treated with diet, regular exercise, and lifestyle changes such as behavioral therapy, as well as medications such as appetite suppressants and fat absorption inhibitors. Since obesity is a chronic disease, long-term use is required when drug treatment is attempted. Currently, products approved for long-term use of more than 3 months in Korea include sibutramine, an appetite suppressant, and orlistat, a fat absorption inhibitor. However, most of these obesity treatment drugs are psychotropic drugs that act on the central nervous system to control appetite, causing side effects such as headaches and vomiting, or diarrhea due to fat absorption, and there are problems such as concerns about abuse. Therefore, research is needed to develop a material with high stability and excellent anti-obesity effects that can resolve the side effects of the above-mentioned commercially available anti-obesity drugs.

Meanwhile, it is not known whether dog grass is effective in preventing, treating, or improving obesity or obesity-related complications.

Accordingly, the inventors of the present invention confirmed that the dog's paw grass directly suppresses the increase in body weight and body fat, so that it can be used as a composition for preventing, treating, or improving obesity, and so that it can be used for the purpose of preventing, treating, or improving obesity-related complications caused by weight gain.

Republic of Korea Patent No. 10-2092254

The present inventors have endeavored to develop a composition for preventing, treating, or improving obesity or obesity-related complications, which has high stability and excellent anti-obesity effects by directly suppressing weight gain, and which can solve the problems of conventional anti-obesity agents, and as a result, have confirmed the excellent anti-obesity effects of the dogtooth grass extract of the present invention, and have completed the present invention based on this.

Accordingly, the present invention aims to provide a pharmaceutical composition for preventing or treating obesity or obesity-related complications, which contains a dog's-root extract as an active ingredient.

In addition, the present invention aims to provide a food composition for preventing or improving obesity or obesity-related complications, which contains a dog's-root extract as an effective ingredient.

However, the technical problems to be achieved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

In order to achieve the above purpose, the present invention provides a preventive or therapeutic pharmaceutical composition containing a dog's mantle extract as an effective ingredient.

In addition, the present invention provides a preventive or improvement food composition containing a dog's-eye extract as an effective ingredient.

In addition, the present invention provides a feed composition for preventing, treating, or improving obesity, which contains a dog grass extract as an effective ingredient.

In one embodiment of the present invention, the obesity-related complication may be at least one selected from the group consisting of, but is not limited to, type 2 diabetes; insulin resistance syndrome; impaired glucose tolerance; visceral fat syndrome; dyslipidemia; hepatotoxic diseases including drug-induced liver damage, viral liver damage, hepatitis, cirrhosis, liver cancer, or hepatic coma; hypertension; stroke; arteriosclerosis; heart failure; angina pectoris; myocardial infarction; fatty liver; and gallbladder disease.

In another embodiment of the present invention, the dogwood extract may be an extract using one or more solvents selected from the group consisting of water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether, subcritical fluid, and supercritical fluid, but is not limited thereto.

In another embodiment of the present invention, the composition can suppress, but is not limited to, an increase in body weight and body fat.

In another embodiment of the present invention, the composition can inhibit fibrosis of liver tissue or adipose tissue, but is not limited thereto.

In another embodiment of the present invention, the composition can suppress, but is not limited to, an increase in plasma lipid concentration.

In another embodiment of the present invention, the composition can suppress, but is not limited to, an increase in lipid content in liver tissue.

In another embodiment of the present invention, the composition can suppress, but is not limited to, an increase in plasma hepatotoxicity indicators.

In another embodiment of the present invention, the food composition may be, but is not limited to, a health functional food composition.

In addition, the present invention provides a method for preventing or treating obesity or obesity-related complications, comprising a step of administering a dog's-root extract to a subject in need thereof.

In addition, the present invention provides a use of a dog's paw extract for the prevention or treatment of obesity or obesity-related complications.

The present invention also provides a use of a dog's paw extract for the manufacture of a medicament for preventing or treating obesity or obesity-related complications.

The dog's paw extract according to the present invention was confirmed to have the effect of suppressing the increase in body weight and body fat caused by a high-fat diet in a mouse model induced by dietary obesity, and improving oxidative damage. Furthermore, it was confirmed to have the effect of improving abnormal lipids and improving fatty liver, hepatotoxicity, and fibrosis. Therefore, the dog's paw extract is expected to be useful for the development of functional foods and pharmaceuticals for not only improving obesity and body fat, but also preventing, treating, or improving obesity-related complications that may be caused by weight gain.

Figure 1 is a schematic diagram showing the experimental design using a diet-induced obesity-induced mouse model to evaluate the anti-obesity effect of the extract of the dog's paw.
Figure 2a is a drawing comparing the body weight change (left) and weight gain inhibition effect (right) according to the breeding period by measuring the body weights of mice in the normal diet group (ND), high-fat diet group (HFD), and group supplemented with dogtooth violet extract (SV) at weekly intervals for 20 weeks (HFD vs. ND: *p<0.05, **p<0.01, ***p<0.001; HFD vs. SV: #p<0.05, ##p<0.01, ###p<0.001, and the same applies hereinafter).
Figure 2b is a diagram comparing the average daily food intake (left) during the breeding period of mice in the normal diet group (ND), high-fat diet group (HFD), and group supplemented with dogtooth violet extract (SV), and the average energy intake (middle) and the food efficiency ratio (FER) (right), which indicates the weight gain in relation to energy intake, after 20 weeks of feeding.
Figure 2c is a drawing showing the weight of white adipose tissue (WAT) extracted from each part of mice fed a normal diet (ND), high-fat diet (HFD), and SV supplemented with dogtooth violet extract for 20 weeks, and expressed as the weight per 100 g of body weight (Epididymal: epididymis, Perirenal: perirenal, Retroperitoneum: retroperitoneum, Mesentric: mesentery, Visceral: viscera, Subcutaneous: subcutaneous, interscapular WAT: interscapular white fat, interscapular BAT: interscapular brown fat).
Figure 3a is a drawing showing the morphological changes in the epididymal white adipose tissue at week 20 after feeding mice in the normal diet group (ND), high-fat diet group (HFD), and group supplemented with dogtooth violet extract (SV) for 20 weeks (upper: H&E staining/lower: Masson's trichrome staining).
Figure 3b is a drawing showing the morphological changes in liver tissue at week 20 after feeding mice in the normal diet group (ND), high-fat diet group (HFD), and group supplemented with dogtooth violet extract (SV) for 20 weeks (upper: H&E staining/lower: Masson's trichrome staining).
Figure 4 is a diagram comparing the plasma neutral lipid (left) and total cholesterol (right) concentrations by week, measured at 4-week intervals for 20 weeks for mice in the normal diet group (ND), high-fat diet group (HFD), and group supplemented with dogtooth violet extract (SV).
Figure 5 is a drawing comparing the weight of liver tissue (upper left), lipid metabolism enzyme activity (upper right), and lipid content (lower; FA: free fatty acid, TG: neutral lipid, CHOL: cholesterol) in mice fed a normal diet (ND), high-fat diet (HFD), and SV supplemented with dogtooth violet extract (SV) for 20 weeks.
Figure 6a is a drawing comparing the lipid peroxide (TBARS) content of red blood cells and liver tissues of mice in the normal diet group (ND), high-fat diet group (HFD), and group supplemented with dogtooth grass extract (SV) after feeding them diets for 20 weeks.
Figure 6b is a diagram comparing the glutathione (GSH) content in the plasma and liver tissue of mice fed a normal diet (ND), a high-fat diet (HFD), and a group supplemented with dogtooth violet extract (SV) for 20 weeks.
Figure 7 is a drawing comparing plasma hepatotoxicity indices in mice fed a normal diet group (ND), a high-fat diet group (HFD), and a group supplemented with dogtooth grass extract (SV) for 20 weeks.

The present invention provides a pharmaceutical composition for preventing or treating obesity or obesity-related complications, comprising a dog's-root extract as an active ingredient.

The “puppy grass ( Setaria viridis )” of the present invention is an annual herb of the Gramineae family of the Poaceae order of monocotyledons, and because its appearance resembles a puppy’s tail, it is also called dog’s tail grass, and in Chinese characters it is called gumicho. There are various types of puppy grass, such as sea dog grass and water dog grass, and they are distributed nationwide, and they grow abundantly in fields, wastelands along roadsides, etc. In oriental medicine, the whole plant is harvested in the summer, dried, and used for medicinal purposes, and it is also mixed with rice or barley to cook rice or make porridge.

In the present invention, the "extract" includes the extract itself and all formulations that can be formed using the extract, such as an extract obtained by extracting the dog grass, a diluted or concentrated extract, a dried product obtained by drying the extract, a controlled or purified product of the extract, or a mixture thereof.

In the dog grass extract of the present invention, the method for extracting the dog grass is not particularly limited, and extraction can be performed according to a method commonly used in the relevant technical field. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, and reflux extraction, and these can be performed alone or in combination of two or more methods.

In the present invention, the type of extraction solvent used to extract the dog's paw grass is not particularly limited, and extraction can be performed using a conventional solvent under conditions of conventional temperature and pressure according to a conventional method known in the art for extracting extracts from natural materials. For example, the dog's paw grass extract in the present invention can be extracted with one or more solvents selected from the group consisting of water, organic solvents having 1 to 6 carbon atoms, and subcritical or supercritical fluids. The organic solvents having 1 to 6 carbon atoms may be one or more selected from the group consisting of alcohols having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, and petroleum ether, but is not limited thereto. The dog's paw grass extract of the present invention can preferably be extracted with ethanol.

The above-mentioned manufactured extract can then be filtered, concentrated, or dried to remove the solvent, and filtration, concentration, and drying can all be performed. For example, filtration can be performed using filter paper or a vacuum filter, concentration can be performed using a vacuum concentrator, and drying can be performed using a freeze-drying method, but is not limited thereto.

In this specification, “active ingredient” means an ingredient that exhibits the intended activity alone or can exhibit the intended activity together with a carrier or the like that is inactive on its own.

In one experimental example of the present invention, in order to evaluate the anti-obesity effect of a dog's paw extract, an experiment was conducted using a mouse model induced with dietary obesity. Specifically, the mice were divided into a normal diet group (ND), a high-fat diet group (HFD), and a dog's paw extract-supplemented group (SV), and were raised for 20 weeks. The body weights were measured. As a result, it was confirmed that the weight gain of high-fat diet-induced obesity mice was suppressed by feeding the dog's paw extract (see Experimental Example 3-1). After raising for 20 weeks, various white adipose tissues were separated from the mice in each group, the weights were measured, and the comparison results showed that the dog's paw extract could also suppress the increase in body fat mass, thereby confirming that the dog's paw extract is effective in weight loss (see Experimental Example 3-2).

In another experimental example of the present invention, the results of morphological analysis of the adipose tissue and liver tissue of mice in each group confirmed that fibrosis of the liver tissue and adipose tissue was suppressed in the SV group compared to the HFD group (see Experimental Example 4).

In another experimental example of the present invention, blood was collected in the same manner at the time of sacrifice of mice in each group and the plasma lipid concentration was analyzed. As a result, it was confirmed that the SV group could suppress the increase in plasma lipid concentration compared to the HFD group (see Experimental Example 5).

In another experimental example of the present invention, liver tissues were extracted from mice of each group and compared and analyzed for liver tissue weight, lipid metabolism enzyme activity, and free fatty acid, neutral lipid, and cholesterol concentrations. As a result, it was confirmed that the SV group could suppress the increase in liver tissue lipid content compared to the HFD group (see Experimental Example 6).

In another experimental example of the present invention, when analyzing the antioxidant-related biomarkers, such as lipid peroxides (TBARS) and glutathione (GSH), from red blood cells and liver tissues collected at the time of sacrifice of mice in each group, it was confirmed that the TBARS content decreased in the SV group compared to the HFD group (see Experimental Example 7-1), and that the GSH content increased in the SV group compared to the HFD group (see Experimental Example 7-2).

In another experimental example of the present invention, when the blood collected at the time of sacrifice of mice in each group was analyzed for plasma hepatotoxicity indicators, GOT (glutamic oxaloacetic transaminase) and GPT (glutamic pyruvic transaminase), it was confirmed that the SV group showed significantly lower plasma GOT and GPT than the HFD group (see Experimental Example 8).

Therefore, it was found through the experimental results of the present invention that the dog's paw extract has an effect of suppressing body weight and body fat increase. In addition, the dog's paw extract of the present invention exhibits an effect of improving abnormal lipids by suppressing an increase in plasma lipid concentration, and has an effect of improving fatty liver and hepatotoxicity by suppressing an increase in liver tissue lipid content and plasma hepatotoxicity indicators, GOT and GPT. In addition, it was found that it has an effect of improving oxidative damage caused by adipocytes and an effect of improving fibrosis by suppressing an increase in TBARS, an antioxidant-related biomarker, and increasing GSH.

Accordingly, it is expected that the extract of dog's paw grass can be applied to the prevention, improvement, or treatment of obesity and various obesity-related complications.

In the present invention, “obesity” refers to a state in which fat cells in the body proliferate and differentiate due to metabolic disorders, resulting in excessive fat accumulation, and may cause related complications including metabolic syndrome accompanied by hypertension, diabetes, and dyslipidemia. When energy intake increases relatively compared to energy consumption, the number and volume of fat cells increase, resulting in an increase in the mass of fat tissue.

The above obesity-related complication may be at least one selected from the group consisting of type 2 diabetes; insulin resistance syndrome; impaired glucose tolerance; visceral fat syndrome; dyslipidemia; hepatotoxic diseases including drug-induced liver damage, viral liver damage, hepatitis, cirrhosis, liver cancer, or hepatic coma; hypertension; stroke; arteriosclerosis; heart failure; angina pectoris; myocardial infarction; fatty liver; and gallbladder disease, and according to a specific experimental example of the present invention, it may be, but is not limited to, dyslipidemia; hepatotoxic diseases including drug-induced liver damage, viral liver damage, hepatitis, cirrhosis, liver cancer, or hepatic coma; or fatty liver.

In the present invention, “prevention” means any action that suppresses or delays the onset of a target disease, and “treatment” means any action that improves or beneficially changes a target disease and its metabolic abnormality symptoms by administering a pharmaceutical composition according to the present invention.

The pharmaceutical composition according to the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions. The excipients may be, for example, at least one selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, moisturizers, film-coating materials, and controlled-release additives.

The pharmaceutical composition according to the present invention may be formulated and used in the form of external preparations such as powders, granules, sustained-release granules, enteric-coated granules, liquids, eye drops, ellipses, emulsions, suspensions, alcohols, troches, aromatic waters, limonades, tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, irrigants, ointments, pastes, sprays, inhalants, patches, sterile injection solutions, or aerosols, and the external preparations may have formulations such as creams, gels, patches, sprays, ointments, ointments, lotions, liniments, pastes, or cataplasmas.

Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

When formulating, it is usually prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants.

The additives of the tablets, powders, granules, capsules, pills and troches according to the present invention include excipients such as corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium monohydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethyl cellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methyl cellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, refined shellac, starch starch, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinyl alcohol, polyvinyl pyrrolidone, and binders such as hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, Disintegrants such as carboxymethyl cellulose calcium, calcium citrate, sodium lauryl sulfate, anhydrous silicic acid, 1-hydroxypropyl cellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, baking soda, polyvinyl pyrrolidone, calcium phosphate, gelled starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethyl cellulose, sucrose, magnesium aluminum silicate, di-sorbitol solution, and light anhydrous silicic acid; Lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium dentata, kaolin, petrolatum, sodium stearate, cocoa butter, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogenated soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, anhydrous silicic acid, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and light anhydrous silicic acid can be used.

Additives that can be used in the liquid formulation according to the present invention include water, diluted hydrochloric acid, diluted sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, ammonia water, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethylcellulose, sodium carboxymethylcellulose, etc.

The syrup according to the present invention may use a solution of white sugar, other sugars or sweeteners, and may also use a flavoring agent, a coloring agent, a preservative, a stabilizer, a suspending agent, an emulsifier, a viscosity increasing agent, etc., as needed.

Purified water may be used in the emulsion according to the present invention, and an emulsifier, preservative, stabilizer, fragrance, etc. may be used as needed.

The suspension according to the present invention may use suspending agents such as acacia, tragacanth, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, and the like. Surfactants, preservatives, stabilizers, colorants, and fragrances may also be used as needed.

The injection according to the present invention includes: solvents such as distilled water for injection, 0.9% sodium chloride injection, Ringer's injection, dextrose injection, dextrose + sodium chloride injection, PEG, lactated Ringer's injection, ethanol, propylene glycol, nonvolatile oils such as sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; solubilizers such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethyl acetamide, butazolidine, propylene glycol, Tween, nitrile acid amide, hexamine, and dimethyl acetamide; buffers such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptone, and gums; It may include isotonic agents such as sodium chloride; stabilizers such as sodium bisulfite (NaHSO ₃ ), carbon dioxide gas, sodium metabisulfite (Na ₂ S ₂ O ₃ ), sodium sulfite (Na ₂ SO ₃ ), nitrogen gas (N ₂ ), and ethylenediaminetetraacetic acid; sulfating agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetic acid disodium, and acetone sodium bisulfite; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium cisplatinum, sodium alginate, Tween 80, and aluminum monostearate.

The suppository according to the present invention comprises cocoa butter, lanolin, witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, subanal, cottonseed oil, peanut oil, palm oil, cocoa butter + cholesterol, lecithin, ranette wax, glycerol monostearate, Tween or Span, Imhausen, monolene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, hexalide base 95, Cotomar, Hydroxocote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Mechanisms such as Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), Suppository type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wecovi (W, R, S, M, Fs), Tezester triglyceride basis (TG-95, MA, 57) can be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations are prepared by mixing the extract with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives.

Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, the "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dosage level can be determined according to the type and severity of the patient's disease, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the excretion rate, the treatment period, the concurrently used drugs, and other factors well known in the medical field.

The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects by taking all of the above factors into consideration, and this can be easily determined by a person skilled in the art to which the present invention belongs.

The pharmaceutical composition of the present invention can be administered to a subject by various routes. All modes of administration can be envisaged, for example, oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, intrathecal injection, sublingual administration, buccal mucosa administration, rectal insertion, vaginal insertion, ocular administration, otic administration, nasal administration, inhalation, spraying through the mouth or nose, skin administration, transdermal administration, etc.

The pharmaceutical composition of the present invention is determined according to the type of drug as an active ingredient along with various related factors such as the disease to be treated, route of administration, age, sex, weight of the patient, and severity of the disease.

In addition, the present invention provides a food composition for preventing or improving obesity or obesity-related complications, comprising a dog's-eye extract as an effective ingredient.

In the present invention, the food composition may be a health functional food composition, but is not limited thereto.

In the present invention, “improvement” means any action of reducing a parameter related to a target disease, for example, the degree of symptoms, by administering a composition according to the present invention. In this case, the health functional food composition may be used before or after the onset stage of the disease, simultaneously with or separately from a drug for treatment, in order to prevent or improve obesity or obesity-related complications.

In the present invention, the "health functional food composition" is characterized in that it is formulated with one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid formulations, including at least one of a carrier, a diluent, an excipient, and an additive.

When the dog's paw extract of the present invention is used as a food additive, the dog's paw extract can be added as it is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The amount of the active ingredient mixed can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing food or beverage, the dog's paw extract of the present invention can be added in an amount of 15 wt% or less, or 10 wt% or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount can be below the above range, and since there is no problem in terms of safety, the active ingredient can also be used in an amount above the above range.

There are no special restrictions on the types of the above foods. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the conventional sense.

The health beverage composition according to the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, like conventional beverages. The natural carbohydrates mentioned above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, a natural sweetener such as thaumatin or stevia extract, or a synthetic sweetener such as saccharin or aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01-0.20 g, or about 0.04-0.10 g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These components may be used independently or in combination. The proportion of these additives is not particularly important, but is generally selected in the range of 0.01-0.20 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides a feed composition for preventing, treating, or improving obesity, which contains a dog's-root extract as an effective ingredient. The dog's-root extract can be prepared in the same manner as in the case of the pharmaceutical composition or food composition.

Specifically, the feed composition containing the dog grass extract according to the present invention as an effective ingredient can be manufactured into various types of feed known in the art, and preferably, it can include concentrated feed, roughage, or special feed.

Concentrated feed includes seeds and fruits of cereals such as wheat, oats and corn; bran including rice bran, wheat bran and barley bran which are by-products obtained by refining grains; sesame cake which is a by-product obtained by extracting soybeans, sesame seeds, linseed and coconut oil; residual starch which is the main component of starch residue which is the remaining substance after removing starch from sweet potatoes and potatoes; animal feed such as fish meal, fish waste, fish soluble which is the concentrated fresh liquid obtained from fish, meat meal, blood meal, feather meal, skim milk powder, dried whey which is the residue obtained when manufacturing cheese from milk or casein from skim milk; yeast, chlorella and seaweed.

Forage includes raw grass feed such as wild grass, pasture, and green grass; root vegetables such as forage turnips, forage beets, and a type of turnip called luterberger; silage, which is stored feed made by filling a silo with raw grass, green grass crops, and grain and fermenting it with lactic acid; hay made by cutting and drying wild grass and pasture; straw from crops for breeding stock, and leaves of legumes.

Special feeds include mineral feeds such as shells and rock salt, urea feeds such as urea or its derivative diuretic isobutane, feed additives that are added in small amounts to compound feeds to supplement ingredients that are often lacking when only natural feed ingredients are mixed, or to increase the storability of the feed, and dietary supplements.

The feed composition according to the present invention may contain various feed additives.

In this specification, "feed additive" refers to a substance added to feed for various purposes such as nutrient supplementation and improving the digestibility and availability of fiber in feed, improving milk quality, preventing reproductive disorders and improving conception rate, and preventing high temperature stress in summer. The feed additive of the present invention corresponds to a supplementary feed under the Feed Management Act, and may additionally include mineral preparations such as sodium bicarbonate (sodium bicarbonate), bentonite, magnesium oxide, and complex minerals; mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium; vitamins such as carotene, vitamin E, vitamins A, D, E, nicotinic acid, and vitamin B complex; protected amino acid preparations such as methionine and lysine; protected fatty acid preparations such as fatty acid calcium salts; live bacteria such as probiotics (lactic acid bacteria), yeast cultures, and mold fermentations; and yeast agents.

The feed composition according to the present invention is not particularly limited and can be applied to any subject for the purpose of improving overweight or obesity. The subject refers to a mammal, including an animal, for example, a non-primate (for example, a cow, a pig, a horse, a cat, a dog, a rat, and a mouse) and a primate (for example, a monkey, for example, a cynomolgous monkey and a chimpanzee). In another specific example, the subject is a livestock animal (for example, a horse, a cow, a pig, etc.) or a pet animal (for example, a dog or a cat).

The dosage of the feed composition according to the present invention will depend on a number of factors such as the species, size, weight, and age of the individual. In principle, a typical dosage may range from 0.001 to 10 g per individual/day, but is not limited thereto.

In addition, the present invention provides a method for preventing or treating obesity or obesity-related complications, comprising a step of administering a dog's-root extract to a subject in need thereof.

In the present invention, the term "subject" means a subject requiring treatment for a disease, and more specifically, a mammal such as a human or non-human primate, mouse, rat, dog, cat, horse, and cow.

In the present invention, “administration” means providing a predetermined composition of the present invention to a subject by any appropriate method.

In addition, the present invention provides a use of a dog's paw extract for the prevention or treatment of obesity or obesity-related complications.

The present invention also provides a use of a dog's paw extract for the manufacture of a medicament for preventing or treating obesity or obesity-related complications.

Hereinafter, preferred examples are presented to help understand the present invention. However, the following examples are provided only to help understand the present invention more easily, and the content of the present invention is not limited by the following examples.

[Experimental example]

Experimental Example 1. Preparation of Dogweed Extract

Setaria viridis used in the experiment was collected from the mountains of Gyeongsan, Gyeongsangbuk-do in August to September and used for extraction. 1 L of 70% ethanol was added to 100 g of the crushed Setaria viridis sample, and the extraction was repeated three times for 3 hours at 60℃. The extracted liquid was filtered, concentrated under reduced pressure, and freeze-dried. Setaria viridis extract powder was obtained, stored frozen, and used in the manufacture of dietary supplements. The extraction yield was 6.05%.

Experimental example 2. Changes in body weight and fat tissue weight due to dog's paw extract

In order to confirm the effects of consuming the extract of dog's paw, an experiment was designed and conducted using a mouse model induced by dietary obesity, as shown in the schematic diagram in Fig. 1.

2-1. Laboratory animals

Experimental animals were 4-week-old male C57BL/6J mice (JA BIO, Korea) purchased from Jung-Ah Bio. After acclimatization for 1 week by providing a pellet-type diet, the experimental animals were divided into a normal diet group (ND), a high-fat diet group (HFD, high-fat diet, 20% fat, 1% cholesterol), and a high-fat diet group supplemented with setaria viridis extract (SV, HFD + 0.3% (w/w) setaria viridis ethanol extract) using a randomized complete block design. The animals were fed the experimental diet of Experimental Example 2-2 below and raised for 20 weeks. The animal rearing room was maintained under constant conditions with constant temperature (24±2℃), constant humidity (50±5%), and a 12-h photoperiod (6:00 AM to 6:00 PM). The experimental diet and water were provided ad libitum to each animal in an individual cage.

2-2. Experimental diet

The composition of the experimental diets of the ND, HFD, and SV groups is as shown in Table 1. The ND group was a normal diet group and was fed an AIN-76 semisynthetic diet, and the HFD group was fed a high-fat diet containing 20% fat and 1% cholesterol by adding lard and cholesterol to the AIN-76 diet. The group with the addition of the dogtooth grass extract was fed a diet with the dogtooth grass extract added at a dosage of 0.3% (w/w) of the high-fat diet.

Experimental Example 3. Analysis of changes in body weight and fat tissue weight by extract of dog's paw

3-1. Weight changes

In order to determine the effect of intake of a dog's mantle extract on weight gain in a diet-induced obesity mouse model, as shown in Experimental Example 2 above, the mice were divided into ND, HFD, and SV groups and raised for 20 weeks, while their body weight and food intake were measured at weekly intervals.

As a result, as shown in Fig. 2a, it was confirmed that the body weight of the HFD group increased rapidly compared to the ND group, whereas the body weight of the SV group significantly decreased compared to the HFD group from 12 weeks of feeding.

In addition, when the body weight gain over 20 weeks was converted to daily weight gain (g/day), it was confirmed that the weight gain in the SV group was significantly suppressed compared to the HFD group.

Meanwhile, as shown in Fig. 2b, there was no significant difference in the average daily food intake (Food intake) and energy intake (Energy intake) between the SV group and the HFD group, but the food efficiency ratio (FER), which indicates the weight gain for energy intake, significantly increased in the HFD group with a high energy density compared to the ND group with a low energy density, whereas it significantly decreased in the SV group compared to the HFD group. From the above results, it was found that the dogtooth grass extract has an effect of significantly reducing body weight in an environment with the same energy intake.

3-2. Weight of adipose tissue

To analyze the change in adipose tissue weight due to the intake of the extract of Dogtooth Venerein, mice raised for 20 weeks were fasted for 12 hours and anesthetized with isofloran (5 mg/kg body weight, Baxter, USA). The liver, epididymal WAT, perirenal WAT, retroperitoneum WAT, mesentric WAT, subcutaneous WAT, visceral WAT, interscapular WAT, and interscapular BAT were extracted. The extracted adipose tissue of each mouse was rinsed several times in saline (0.9% saline solution) solution, the moisture was removed, the weight was measured, and the result was expressed as the weight per 100 g of body weight for comparison. Furthermore, the extracted tissues were rapidly frozen in liquid nitrogen for further analysis and stored at -70°C until analysis.

As a result, as shown in Fig. 2c, in the case of the HFD group, the weight of adipose tissue in all parts significantly increased compared to the ND group, whereas in the SV group, the weight of epididymal WAT, perirenal WAT, mesentric WAT, visceral WAT, and total WAT significantly decreased compared to the HFD group. Through the above results, it was found that the dogtooth grass extract has a body fat reduction effect by significantly reducing fat mass.

Experimental Example 4. Analysis of the effect of dog's mantle extract on morphological changes in liver and adipose tissue

In order to observe the morphological changes in tissues by the extract of the dogtooth root, some of the liver tissue and epididymal white adipose tissue extracted in the above Experimental Example 3-2 were fixed in a 10% formaldehyde solution for 24 hours, then exchanged twice with the same solution, dehydrated with doubled volume ethanol, embedded in paraffin, and treated with poly-L-lysine to produce 5 μm thick tissue sections, which were then stained with hematoxylin and eosin (H&E) and observed under a light microscope at a magnification of 200x. In addition, Massons' trichrome staining (connective tissue: blue, nucleus: dark red, cytoplasm: pink) was performed to stain collagen and muscle fibers, and observed under a light microscope at a magnification of 200x.

4-1. Types of adipose tissue

As shown in Fig. 3a, when observing the epididymal white adipose tissue, the size of the fat cells in the HFD group was observed to be larger than that in the ND group, and the size of the fat cells in the SV group was confirmed to be smaller than that in the HFD group.

In addition, as a result of Masson's trichrome staining to determine the effect of the extract of dog's mantle on the fibrosis of adipose tissue, it was observed that more fibrous connective tissue was accumulated in the adipocytes of the HFD group compared to the ND group, and the SV group showed suppressed fibrosis of adipose tissue caused by a high-fat diet.

4-2. Morphology of liver tissue

As shown in Fig. 3b, when observing liver tissue, accumulation of lipid droplets was observed in the HFD group compared to the ND group, centered on the portal vein of the liver tissue, whereas the number of lipid droplets in the SV group was found to be reduced compared to the HFD group.

In addition, as a result of Masson's trichrome staining to determine the effect of the extract of the dogtooth grass on fibrosis of liver tissue, it was observed that a lot of connective tissue was accumulated around the portal vein in the HFD group, while fibrosis of liver tissue caused by a high-fat diet was suppressed in the SV group.

Experimental Example 5. Analysis of the effect of dog's mantle extract on plasma lipid concentration

5-1. Plasma neutral lipid and total cholesterol concentrations by week

In order to confirm the effect of ingestion of the extract of the dog's paw on plasma lipid concentration, the mice of Experimental Example 2 were fasted for 12 hours every 4 weeks during the breeding period, and then tail blood was collected without anesthesia to measure plasma neutral lipid and total cholesterol concentrations.

As a result, as shown in Fig. 4, in the case of plasma triglyceride concentration, the plasma triglyceride concentration of the ND group was significantly lower than that of the HFD group in the 4th week of experimental diet feeding, but there was no significant difference between the groups during the subsequent experimental period. Meanwhile, in the case of plasma total cholesterol concentration, the plasma total cholesterol concentration of the HFD group was significantly higher than that of the ND group from the 4th week of experimental diet feeding, and the plasma total cholesterol concentration of the SV group was significantly lower than that of the HFD group from the 8th week of experimental diet feeding.

5-2. Plasma lipid concentration

In the above Experimental Example 3-2, blood was collected from the inferior vena cava of the abdomen before adipose tissue extraction. The collected blood was centrifuged at 3,000 rpm, 4°C for 15 minutes, and plasma was collected, which was then stored at -70°C until sample analysis. The concentrations of plasma triglycerides, total cholesterol, free fatty acids, apolipoprotein A-1 (Apo A-1), and apolipoprotein B (Apo B) were compared from the stored plasma. At this time, plasma neutral lipid and total cholesterol concentrations were measured using an enzyme kit from Asan Pharm Co., Seoul, Korea, and free fatty acid content was measured using a free fatty acid measurement solution (Non-Esterified fatty acid, NEFA kit, Shin Yang Chemical) that utilizes a colorimetric method using an enzyme method, and plasma apolipoprotein A-1 (Apo A-1) and apolipoprotein B (Apo B) concentrations were measured using an Apo A-1 and Apo B measurement kit (Nitto Soko Co., Ltd., Tokyo, Japan).

Measurements Experimental groups ND HFD SV Free fatty acid (mol/L) 1.12±0.04 1.08±0.02 0.94±0.03 ^## Triglyceride (mmol/L) 1.00±0.10 0.95±0.05 0.79±0.02 ^# Total cholesterol (mmol/L) 4.09±0.30 7.17±0.65 ^** 5.68±0.37 Apo A-1 (mg/ml) 0.28±0.02 0.34±0.03 0.35±0.01 Apo B (mg/ml) 0.21±0.01 0.40±0.05 ^* 0.26±0.03 ^# Apo B / Apo A-1 0.77±0.06 1.15±0.10 ^** 0.73±0.06 ^##

As a result, as shown in Table 2, it was confirmed that the concentrations of plasma free fatty acids, neutral lipids, and Apo B in the SV group significantly decreased compared to the HFD group. Therefore, from the above results, it was found that the extract of Dogtooth vulgaris had an effect of improving abnormal lipids by reducing plasma lipid components.

Experimental Example 6. Analysis of the effect of dog's paw extract on liver tissue weight and lipid concentration

6-1. Weight of liver tissue

As a result of measuring the weight of liver tissue using the method of the above Experimental Example 3-2 and comparing it by expressing it as the weight per 100 g of body weight, as shown in Fig. 5, in the case of the HFD group, hypertrophy was observed in which the weight of liver tissue increased rapidly compared to the ND group, whereas in the case of the SV group, it was confirmed that it significantly decreased compared to the HFD group.

6-2. Activity of lipid metabolism enzyme (Phosphatidate phosphohydrolase) in liver tissue

To isolate enzyme sources from liver tissue, the separation method performed by Hulcher et al. (1973) was applied with some modifications. In the above Experimental Example 3-2, 0.5 g of frozen liver tissue was added with a buffer solution containing 0.1 M triethanolamine, 0.02 M ethylenediamine tetracetate (EDTA, pH 7.4), and 0.002 M dithiothreitol (DTT), and homogenized with a glass Teflon homogenizer (Glascol, 099C K44, USA) in an ice-cold state, centrifuged at 3,000 rpm at 4°C for 15 minutes, and only the supernatant was centrifuged again at 13,000 rpm at 4°C for 15 minutes. Among these, the lower layer sediment separated from the supernatant was used as the mitochondria fraction, and the cytosol fraction of the supernatant was obtained using an ultracentrifuge (Beckman, Optima TLX-120, USA) at 32,500 rpm, 4℃ for 1 hour. The microsome fraction was prepared by adding the same buffer solution to the separated pellet from the cytosol fraction of the supernatant, ultracentrifuging again at 33,000 rpm, 4℃ for 40 minutes, and then dissolving the pellet in 1 ml of buffer solution, storing it at -70℃, and using it for analysis and protein quantification.

Phosphatidate phosphohydrolase (PAP) activity was measured according to the method of Walton et al. (1985). That is, 50 μL of substrate containing 1 mM phosphatidate and phosphatidylcholine dissolved in 0.9% NaCl solution was added to 50 μL of the reaction solution with and without addition of 0.05 M Tris-HCl (pH 7.0), 1.25 mM Na ₂ -EDTA, and 1 mM MgCl ₂ , and then 0.1 mL of the microsome fraction was added to initiate the reaction. After reacting at 37°C for 15 minutes, 0.1 mL of 1.8 MH ₂ SO ₄ was added to stop the reaction, and 0.1 mL of 0.13% sodium dodecyl sulfate solution and 0.25 mL each of 1.25% ascrobic acid and 0.32% ammonium molybdate were added. After reacting and developing color at 45°C for 20 minutes, the absorbance was measured at 820 nm.

As a result, as shown in Figure 5, the activity of PAP, an enzyme related to neutral lipid synthesis in liver tissue, significantly increased in the HFD group compared to the ND group, but significantly decreased in the SV group compared to the HFD group.

6-3. Liver tissue lipid content

In order to measure the lipid content of liver tissue, the lipid of the liver tissue stored in the above Experimental Example 3-2 was extracted according to the method of Folch et al. (1957), and then the extract was volatilized with nitrogen gas at 37°C and diluted with isopropanol. For quantification, the lipid component was extracted by mixing 3 mM cholic acid as an emulsifier with the enzyme reagent and 0.5% Triton X-100 to remove the turbidity that occurs during color development, and then quantified in the same way as the plasma neutral lipid, cholesterol, and free fatty acid quantification method.

As a result, as shown in Fig. 5, it was confirmed that the liver tissue lipid content significantly increased in the HFD group compared to the ND group, whereas the liver tissue lipid content significantly decreased in the SV group compared to the HFD group. Therefore, it was found from the above results that the dogtooth grass extract had an effect on improving fatty liver.

Experimental Example 7. Analysis of the effect of dog grass extract on antioxidant-related biomarkers

To determine the effect of the extract of dog's mantle on oxidative damage induced by a high-fat diet, lipid peroxide and glutathione were measured and compared.

7-1. Lipid peroxides of red blood cells and liver tissue (TBARS; Thiobarbituricacid reactive substance)

The measurement of erythrocyte lipid peroxide content was performed using the method of Tarladgis et al. (1964). Erythrocytes were separated according to the method of McCord and Fridovich (1969). Specifically, the heparinized blood of Experimental Example 5 was centrifuged at 3,000 rpm and 4°C for 15 minutes to completely remove plasma and buffy coat, and then washed three times with 0.9% saline solution to obtain erythrocytes. 50 uL of the obtained erythrocytes were added with 3 mL of 5% triochloroacetic acid (TCA) and 1 mL of 0.06 M thiobarbituric acid (TBA) and reacted at 80°C for 90 minutes. After cooling to room temperature, the mixture was centrifuged at 2,000 rpm and 25°C for 15 minutes, and the supernatant was collected to measure the absorbance at 535 nm. Lipid Peroxidation (MDA) standard solution was prepared by dissolving 1 mmol of TMP in 100 mL of 0.01 N HCL solution, reacting at 50°C for 60 minutes, and then cooling to room temperature to hydrolyze TMP into MDA. Then, 1 mL of the hydrolyzed TMP solution was diluted in 100 mL of 0.01 M Na ₃ PO ₄ (pH 7.0) buffer to prepare an MDA standard solution (1 x 10 ^-4 M). The absorption spectrum of the MDA standard solution was obtained at 267 nm, and the exact concentration was calculated from the extinction coefficient for correction, after which a TBA-MDA chromopore standard curve was obtained, and the amount of TBA reactant was calculated in terms of the MDA extinction coefficient from this curve.

Furthermore, the lipid peroxide content of liver tissue was determined using the method of Ohkawa et al. (1979). 0.5 g of the liver tissue stored frozen in Experimental Example 3-2 was added to a buffer solution containing 0.1 M triethanolamine, 0.02 M EDTA (pH 7.4), and 0.002 M DTT, and homogenized with a glass Teflon homogenizer (Glascol, 099C K44, USA) in an ice-cold state. 0.2 mL of the homogenate, 0.2 mL of 8.1% sodium dodecyl sulfate (SDS) solution, and 0.6 mL of distilled water were mixed, left at room temperature for 5 minutes, and then 1.5 mL of 20% acetate (pH 3.5) buffer and 1.5 mL of 0.8% TBA were added, and the mixture was reacted at 95°C for 1 hour. After the reaction, the sample was cooled to room temperature, 1 mL of distilled water and 5 mL of n-butanol:pyridine (15:1) solution were added, and the sample was centrifuged at 3,000 rpm and 20°C for 15 minutes. The absorbance of the supernatant was measured at 532 nm. The MDA standard solution hydrolyzed TMP, and the amount of TBA reactant produced at 267 nm was calculated as the MDA absorption coefficient.

As a result, as shown in Fig. 6a, it was confirmed that the lipid peroxide levels in red blood cells and liver tissue significantly increased in the HFD group compared to the ND group, but significantly decreased in the SV group compared to the HFD group.

7-2. Glutathione (GSH) in plasma and liver tissue

Glutathione content includes both oxidized GSH and reduced GSH. Liver tissue glutathione content was measured by modifying and supplementing the method of Fiala et al. (1976). In the above Experimental Example 3-2, 0.5 g of liver tissue stored frozen was homogenized with a buffer solution containing 0.1 M triethanolamine, 0.02 M EDTA (pH 7.4), and 0.002 M DTT in an ice-cold state using a glass Teflon homogenizer (Glascol, 099C K44, USA). 0.3 mL of distilled water and 0.5 mL of 4% sulfosalicylic acid were added to 0.2 mL of the homogenate, and centrifuged at 25,000 rpm, 4°C for 10 minutes to obtain the supernatant. 2.7 mL of 0.1 M disulfide reagent (5.5'-dithiobis + 0.1 M sodium phosphate buffer, pH 8.0) was added to 0.3 mL of the supernatant, and the mixture was reacted at room temperature for 20 minutes, and the absorbance was measured at 412 nm.

Furthermore, the plasma glutathione content was measured by hemolysing the red blood cells obtained in Experimental Example 7-1 with an equal amount of distilled water.

As a result, as shown in Fig. 6b, it was confirmed that the glutathione content of red blood cells and liver tissue significantly decreased in the HFD group compared to the ND group, but significantly increased in the SV group compared to the HFD group.

Experimental Example 8. Analysis of the effect of dog's mantle extract on liver tissue toxicity

In order to confirm the effect of the dog's paw extract on hepatocyte damage, GOT (glutamic oxaloacetic transaminase) and GPT (glutamic pyruvic transaminase), which are closely related to hepatocyte damage, were measured and compared. Specifically, the activities of GOT and GPT were measured using an enzyme kit from Asan Pharm Co., Seoul, Korea, using plasma GOT and GPT from the blood collected in Experimental Example 5-2.

As a result, as shown in Fig. 7, the HFD group showed significantly increased plasma GOT and GPT contents compared to the ND group, but the SV group showed significantly decreased plasma GOT and GPT contents compared to the HFD group. From the above results, it can be seen that the extract of Dogtooth vulgaris has an effect of inhibiting hepatotoxicity.

Experimental Example 9. Statistical Analysis

All experimental results were calculated using the SPSS package program versuib 25.0 (Statistical Paskage for the Social Sciences, SPSS Inc., Chicago), one of the computer statistics programs. Student's t-test was performed to test the significance between the ND group and the HFD group and between the HFD group and the SV group. All results were expressed as the mean ± S.E.

The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential characteristics of the present invention. Therefore, it should be understood that the embodiments described above are exemplary in all respects and not restrictive.

Claims (17)

A pharmaceutical composition for preventing or treating complications due to obesity, comprising an extract of dog's mantle as an active ingredient,
A pharmaceutical composition for preventing or treating complications due to obesity, characterized in that the complication due to obesity is liver cirrhosis.
delete In the first paragraph,
A pharmaceutical composition, characterized in that the above-mentioned dogwood extract is an extract using at least one solvent selected from the group consisting of water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether, subcritical fluid, and supercritical fluid.
In the first paragraph,
A pharmaceutical composition characterized in that the composition suppresses an increase in body weight and body fat.
In the first paragraph,
A pharmaceutical composition characterized in that the composition inhibits fibrosis of liver tissue or adipose tissue.
In the first paragraph,
A pharmaceutical composition characterized in that the composition suppresses an increase in plasma lipid concentration.
In the first paragraph,
A pharmaceutical composition characterized in that the composition suppresses an increase in lipid content in liver tissue.
In the first paragraph,
A pharmaceutical composition characterized in that the composition suppresses an increase in plasma hepatotoxicity indicators.
A food composition for preventing or improving complications caused by obesity, containing an extract of dog grass as an effective ingredient.
A food composition for preventing or improving complications due to obesity, characterized in that the complication due to obesity is liver cirrhosis.
delete In Article 9,
A food composition, characterized in that the above-mentioned dogwood extract is an extract using at least one solvent selected from the group consisting of water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether, subcritical fluid, and supercritical fluid.
In Article 9,
A food composition characterized in that the composition suppresses an increase in body weight and body fat.
In Article 9,
A food composition characterized in that the composition inhibits fibrosis of liver tissue or adipose tissue.
In Article 9,
A food composition characterized in that the composition suppresses an increase in plasma lipid concentration.
In Article 9,
A food composition characterized in that the composition suppresses an increase in lipid content in liver tissue.
In Article 9,
A food composition characterized in that the composition suppresses an increase in plasma hepatotoxicity indicators.
In Article 9,
A food composition, characterized in that the above food composition is a health functional food composition.