WO2024117763A1 - Pharmaceutical composition for preventing or treating fatty liver or obesity, comprising extract of crataegus pinnatifida bunge roots as active ingredient - Google Patents

Pharmaceutical composition for preventing or treating fatty liver or obesity, comprising extract of crataegus pinnatifida bunge roots as active ingredient Download PDF

Info

Publication number
WO2024117763A1
WO2024117763A1 PCT/KR2023/019426 KR2023019426W WO2024117763A1 WO 2024117763 A1 WO2024117763 A1 WO 2024117763A1 KR 2023019426 W KR2023019426 W KR 2023019426W WO 2024117763 A1 WO2024117763 A1 WO 2024117763A1
Authority
WO
WIPO (PCT)
Prior art keywords
preventing
fatty liver
obesity
composition
root extract
Prior art date
Application number
PCT/KR2023/019426
Other languages
French (fr)
Korean (ko)
Inventor
남주옥
이은비
채종범
Original Assignee
경북대학교 산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR1020220165578A external-priority patent/KR20240081878A/en
Priority claimed from KR1020220165566A external-priority patent/KR20240081872A/en
Application filed by 경북대학교 산학협력단 filed Critical 경북대학교 산학협력단
Publication of WO2024117763A1 publication Critical patent/WO2024117763A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/734—Crataegus (hawthorn)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/06—Preparations for care of the skin for countering cellulitis

Definitions

  • the present invention relates to a composition for preventing, improving or treating fatty liver disease or obesity, including a hawthorn ( Crataegus pinnatifida ) extract as an active ingredient.
  • a hawthorn Crataegus pinnatifida
  • Obesity generally refers to the state of excessive fatty tissue in the body, and is a phenomenon in which the energy consumed through food is not balanced with the energy consumed through physical activity, and the excess energy is accumulated as body fat.
  • Obesity has a wide variety of causes, including genetic factors, environmental influences due to irregular eating habits, lack of exercise or westernized eating habits, depression, boredom, psychological effects due to excessive stress, and pathological factors due to changes in thyroid or adrenal cortex hormones. It is known to be caused by Recently, with economic growth and changes in lifestyle, there have been many changes in eating habits, and busy modern people are increasingly overweight and obese due to high-calorie diets such as fast food and little exercise.
  • adipocyte differentiation may be helpful in treating obesity.
  • anti-obesity studies related to the inhibition of adipocyte differentiation and lipid accumulation have been reported, and 3T3-L1 mouse fibroblasts are a well-characterized model for the differentiation of preadipocytes and are generally used for in vitro studies. .
  • liver is the most important organ in our body related to metabolism, detoxification, and excretion. Liver function deteriorates due to viruses, alcohol, drugs, malnutrition, obesity, etc. In particular, fatty liver caused by high sugar and high fat intake in modern people can further progress to steatohepatitis.
  • Fatty liver is divided into alcoholic fatty liver disease and non-alcoholic fatty liver disease. If the weight increases by more than 5% due to fat accumulation not caused by alcohol, it is defined as non-alcoholic fatty liver disease.
  • Nonalcoholic fatty liver disease which is unrelated to drinking, is reported in as few as 6.3% and as many as 33% worldwide, and non-alcoholic fatty liver lesions are reported to occur even in a small number of normal-weight adults.
  • Non-alcoholic fatty liver disease is caused by increased absorption of free fatty acids from adipose tissue into the liver or increased biosynthesis of neutral fat in the liver. In particular, when insulin resistance occurs, lipolysis is promoted in adipose tissue, increasing fatty acids flowing into the liver. It causes impaired glucose utilization in muscle cells, increasing hyperinsulinemia and carbohydrate conversion to fat (de novo lipogenesis), further increasing fat accumulation in the liver.
  • Non-alcoholic fatty liver disease caused by obesity progresses from simple fatty liver (steatosis) to non-alcoholic steatohepatitis (non-alcoholic steatohepatitis) in severe cases, as neutral fat accumulates in the body. Unlike other diseases, it progresses to cirrhosis, loss of liver function, etc. without general symptoms. Early treatment and prevention are necessary because it causes complex and progressive liver damage such as liver cancer.
  • Obesity can be treated through diet, regular exercise, lifestyle changes such as behavioral therapy, and medications such as appetite suppressants and fat absorption inhibitors. Since obesity is a chronic disease, attempting drug treatment requires long-term use. Currently, products approved for long-term use in Korea for more than 3 months include the appetite suppressant sibutramine and the lipolytic enzyme inhibitor orlistat. ). However, most of these obesity treatment drugs are psychotropic drugs that act on the central nervous system to control appetite, so they come with side effects such as headaches and vomiting and have problems such as concerns about abuse. Therefore, research is being actively conducted to develop materials with high stability and excellent anti-obesity effect that can solve the side effects of the commercially available anti-obesity drugs.
  • Hawthorn (Crataegus pinnatifida) is a deciduous tree belonging to the Rosaceae family of the rose order. In Korea, it grows wild in mountainous regions across the country. Its fruits have been used for food and medicinal purposes since ancient times, but not much is known about the physiological effects of the roots of hawthorn.
  • the present inventors completed the present invention by confirming the fatty liver improvement effect or anti-obesity activity of the hawthorn root extract.
  • the present inventors have made efforts to develop a composition for preventing or treating fatty liver or obesity with high stability that can solve the problems of conventional anti-obesity drugs and excellent anti-fatty liver or anti-obesity effects.
  • the hawthorn root extract of the present invention has been found to be an excellent anti-obesity drug.
  • the present invention was completed after confirming that it had an anti-fatty liver or anti-obesity effect.
  • the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating fatty liver comprising hawthorn root extract as an active ingredient.
  • Another object of the present invention is to provide a food composition for preventing or improving fatty liver disease containing hawthorn root extract as an active ingredient.
  • the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating obesity containing hawthorn root extract as an active ingredient.
  • Another object of the present invention is to provide a food composition for preventing or improving obesity containing hawthorn root extract as an active ingredient, and another object is to provide a cosmetic composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  • a composition is provided.
  • Another object of the present invention is to provide a quasi-drug composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  • the present invention provides a pharmaceutical composition for preventing or treating fatty liver comprising hawthorn root extract as an active ingredient.
  • the present invention provides a food composition for preventing or improving fatty liver disease containing hawthorn root extract as an active ingredient.
  • the present invention provides a method for preventing, improving, or treating fatty liver disease, comprising administering a hawthorn root extract or a composition containing the same as an active ingredient to an individual in need thereof.
  • the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient to prevent, improve, or treat fatty liver.
  • the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient for preparing a preparation for preventing, improving, or treating fatty liver disease.
  • the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient for preparing a medicine for preventing, improving, or treating fatty liver disease.
  • the hawthorn root extract contains water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, and ethyl acetate.
  • one or more solvents selected from the group consisting of acetate, methylene chloride, hexane, cyclohexane, petroleum ether, dichloromethane, subcritical fluids, and supercritical fluids It may be an extract, but is not limited thereto.
  • the fatty liver may be non-alcoholic fatty liver, but is not limited thereto.
  • the composition can improve liver damage induced by a high-fat diet, but is not limited thereto.
  • the composition can reduce serum ALT or AST activity, and preferably, the composition can reduce serum ALT or AST activity in a high-fat diet group, but is not limited thereto.
  • the hawthorn root extract may inhibit the expression of transcription factors or adiponectin involved in adipocyte differentiation and maturation, but is not limited thereto.
  • the transcription factor may be peroxisome proliferator activated receptor gamma (PPAR ⁇ ), sterol regulator binding protein-1 (SREBP-1), or CCAAT-enhancer binding protein alpha (C/EBP ⁇ ).
  • PPAR ⁇ peroxisome proliferator activated receptor gamma
  • SREBP-1 sterol regulator binding protein-1
  • C/EBP ⁇ CCAAT-enhancer binding protein alpha
  • the food composition may be a health functional food composition, but is not limited thereto.
  • the present invention provides a pharmaceutical composition for preventing or treating obesity containing hawthorn root extract as an active ingredient.
  • the present invention provides a food composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  • the present invention provides a cosmetic composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  • the present invention provides a quasi-drug composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  • the present invention provides a method for preventing, improving, or treating obesity, comprising administering a hawthorn root extract or a composition containing the same as an active ingredient to an individual in need thereof.
  • the present invention provides a use of hawthorn root extract or a composition containing it as an active ingredient to prevent, improve, or treat obesity.
  • the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient for preparing a preparation for preventing, improving or treating obesity.
  • the present invention provides the use of a hawthorn root extract or a composition containing it as an active ingredient for preparing a medicine for preventing, improving, or treating obesity.
  • the hawthorn root extract contains water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, and ethyl acetate.
  • one or more solvents selected from the group consisting of acetate, methylene chloride, hexane, cyclohexane, petroleum ether, dichloromethane, subcritical fluids, and supercritical fluids It may be an extract, but is not limited thereto.
  • the hawthorn root extract may have one or more of the following properties, but is not limited thereto:
  • the hawthorn root extract may inhibit the expression of transcription factors or adiponectin involved in adipocyte differentiation and maturation, but is not limited thereto.
  • the transcription factor may be peroxisome proliferator activated receptor gamma (PPAR ⁇ ), sterol regulator binding protein-1 (SREBP-1), or CCAAT-enhancer binding protein alpha (C/EBP ⁇ ).
  • PPAR ⁇ peroxisome proliferator activated receptor gamma
  • SREBP-1 sterol regulator binding protein-1
  • C/EBP ⁇ CCAAT-enhancer binding protein alpha
  • the food composition may be a health functional food composition, but is not limited thereto.
  • hawthorn root extract effectively inhibits adipocyte differentiation without cytotoxicity, reduces lipid accumulation in adipocytes, and inhibits the expression of transcription factors involved in adipocyte differentiation and maturation.
  • the effect of inhibiting hepatic steatosis and improving liver damage caused by a high-fat diet was confirmed in high-fat diet mice, and the inhibition of fat production was confirmed in high-fat diet mice. Therefore, the hawthorn root extract of the present invention can be usefully used as a pharmaceutical composition and health functional food for preventing, treating, or improving fatty liver disease or obesity.
  • Figure 1a shows the results of measuring the viability of 3T3-L1 preadipocytes by MTT assay at 595 nm absorbance. CON was treated with the same amount of DMSO as CRE.
  • Figure 1b shows an image observed after staining 3T3-L1 preadipocytes with Oil Red O solution (left), and the result of measuring the content of Oil Red O (ORO) solution at absorbance at 450 nm (right, *p ⁇ 0.05). It represents.
  • Figure 1c shows the results of measuring the triglyceride content in 3T3-L1 preadipocytes using absorbance at 570 nm. *p ⁇ 0.05
  • Figure 1d shows the results of measuring the mRNA expression levels of Adiponectin, C/EBP ⁇ , PPAR ⁇ , and SREBP-1 in 3T3-L1 preadipocytes. *p ⁇ 0.05
  • Figure 1e shows the results of measuring the protein expression levels of C/EBP ⁇ and PPAR ⁇ in 3T3-L1 preadipocytes. *p ⁇ 0.05
  • Figure 2a is an image showing fat globules in iWAT, eWAT, and liver tissue stained with H&E.
  • Figure 2b shows the results of measuring the protein expression levels of SREBP-1 and PPAR ⁇ in iWAT tissue. *p ⁇ 0.05
  • Figure 2c shows representative images of the liver of each mouse group.
  • Figure 2d shows the results of measuring the plasma levels of AST and ALT in each mouse group. *p ⁇ 0.05
  • Figure 3a is an image of reactive oxygen species in 3T3-L1 preadipocytes observed through a fluorescence microscope.
  • Figure 3b shows the results of quantitative measurement of reactive oxygen species in 3T3-L1 preadipocytes using a flow cytometer.
  • Figure 4a shows the feed intake ratio of each mouse group. **p ⁇ 0.05
  • Figure 4b shows the results of measuring the body weight of mice in each group at weekly intervals. *p ⁇ 0.05
  • Figure 4c shows the results of measuring the weight of adipose tissue and organs in each group (top, *p ⁇ 0.05 and **p ⁇ 0.01) and the results of measuring obesity relative to body weight (bottom).
  • Figure 4d shows the results of measuring the TG content (left) and total cholesterol content (right) of each mouse group. *p ⁇ 0.05
  • Figures 4E to 4G show representative images of abdominal organs (Figure 4E), iWAT ( Figure 4F), and eWAT ( Figure 4G) of each mouse group.
  • the present inventors confirmed that the hawthorn root extract of the present invention has an excellent fatty liver improvement effect and completed the present invention.
  • the present inventors tried to develop a composition for preventing or treating obesity with high stability and excellent anti-obesity effect, and as a result, they confirmed that the hawthorn root extract of the present invention had excellent anti-obesity effect and completed the present invention.
  • the present invention provides a pharmaceutical composition for preventing, improving, or treating fatty liver disease or obesity, comprising hawthorn root extract as an active ingredient.
  • hawthorn (Crataegus pinnatifida for. pinnatifida) used in the present invention is a deciduous small tree belonging to the genus of Hawthorn in the Rosaceae family. In Korea, it grows naturally in mountainous areas across the country, and the leaves, flowers, fruits, bark, and roots of the hawthorn tree have been used since ancient times. It was used for food or medicinal purposes.
  • hawthorn root extract refers to an extract obtained by extraction treatment of the hawthorn root, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a crude product or purified product of the extract, or a mixture thereof. This includes the extract itself and all formulations of extracts that can be formed using the extract.
  • the method of extracting the hawthorn root is not particularly limited, and may be extracted according to a method commonly used in the art.
  • Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, and reflux extraction, which may be performed alone or in combination of two or more methods, but are not limited thereto.
  • the hawthorn root extract is water, alcohol with 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, and methylene. Extraction with one or more solvents selected from the group consisting of methylene chloride, hexane, cyclohexane, petroleum ether, dichloromethane, subcritical fluid, and supercritical fluid. However, it is not limited to this.
  • the type of solvent used to extract hawthorn roots in the present invention is not particularly limited, and is performed according to a conventional method known in the art for extracting extracts from natural products, that is, under conventional conditions of temperature and pressure. It can be extracted using a solvent. For example, it may 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 solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, benzene, chloroform, ethyl acetate, and methylene chloride.
  • the solvent is 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 80%. It may be 100%, 85% to 100%, 90% to 100%, 95% to 100%, or 100% solvent, but is not limited thereto.
  • the solvent is 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 80%. It may be 100%, 85% to 100%, 90% to 100%, 95% to 100%, or 100% methanol, preferably 100% methanol, but is not limited thereto.
  • Conventional extraction methods such as cold immersion, warm immersion, and heating can be used using the above solvent.
  • fatty liver may be non-alcoholic fatty liver, but is not limited thereto.
  • the composition can improve liver damage induced by a high-fat diet, but is not limited thereto.
  • the composition can reduce serum ALT or AST activity, and preferably, the composition can reduce serum ALT or AST activity in the high-fat diet group, but is not limited thereto.
  • hawthorn root extract can inhibit the expression of transcription factors or adiponectin involved in adipocyte differentiation and maturation, but is not limited thereto.
  • the transcription factor may be peroxisome proliferator-activated receptor gamma (PPAR ⁇ sterol regulator binding protein-1 (SREBP-1) or CCAAT-enhancer binding protein alpha (C/EBP ⁇ ), but is limited thereto. no.
  • PPAR ⁇ sterol regulator binding protein-1 SREBP-1
  • C/EBP ⁇ CCAAT-enhancer binding protein alpha
  • the hawthorn root extract may have one or more of the following properties, but is not limited thereto:
  • hawthorn root extract reduces fat content by inhibiting 3T3-L1 preadipocyte differentiation without causing cytotoxicity (see Example 1).
  • hawthorn root extract was confirmed to have an inhibitory effect on steatosis and an improvement in liver damage in HFD-induced obese mice (see Example 2).
  • antioxidant in the present invention refers to the action of suppressing oxidation.
  • the human body has a balance of prooxidants and antioxidants, but this balance can become imbalanced due to various factors. When this is achieved and the tendency is toward promoting oxidation, oxidative stress is induced, causing potential cell damage and pathological diseases.
  • Reactive oxygen species ROS
  • ROS reactive oxygen species
  • Reactive nitrogen species such as NO, HNO 2 , and ONOO-
  • NO reactive nitrogen species
  • HNO 2 oxidize and destroy cells in the body, thereby exposing them to various diseases.
  • hawthorn root extract reduces the content of active oxygen in 3T3-L1 preadipocytes (see Example 3).
  • hawthorn root extract inhibits fat production in HFD-induced obese mice (see Example 4).
  • the term "obesity" of the present invention refers to a state in which fat cells proliferate and differentiate in the body due to metabolic disorders, resulting in excessive accumulation of fat, and is accompanied by high blood pressure, diabetes, and dyslipidemia. It can lead to associated complications, including syndromes. When energy absorption increases relative to consumption, the number and volume of fat cells increase, resulting in an increase in the mass of fat tissue.
  • white adipocytes are cells with the function of accumulating large amounts of fat energy as neutral fat in the body, they proliferate intensively in late pregnancy, infancy, and puberty, and swell up to 15 times, causing the phenomenon of fat hypertrophy. It may also be the cause, and generally, “adipocytes” refer to “white fat cells.”
  • differentiation in the present invention refers to the phenomenon in which cells become specialized in structure or function while they divide and grow, that is, the cells, tissues, etc. of living organisms change their form or function to perform a given task. It says that In general, it is a phenomenon in which a relatively simple system is separated into two or more qualitatively different subsystems. For example, in ontogeny, distinctions such as the head or body arise between parts of an egg that were initially homogeneous, or distinctions such as muscle cells or nerve cells develop in cells, so qualitative changes occur between parts of a biological system that were initially almost homogeneous. The occurrence of personal differences, or the state of being divided into qualitatively distinguishable sub-regions or subsystems as a result, is called differentiation.
  • the content of the hawthorn root extract in the composition of the present invention can be appropriately adjusted depending on the symptoms of the disease, the degree of progression of the symptoms, the patient's condition, etc., for example, 0.0001 to 99.9% by weight, or 0.001 to 50%, based on the total weight of the composition. It may be weight percent, but is not limited thereto.
  • the content ratio is a value based on the dry amount with the solvent removed.
  • the pharmaceutical composition according to the present invention may further include appropriate carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions.
  • the excipient may be, for example, one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, humectants, film-coating materials, and controlled-release additives.
  • the pharmaceutical composition according to the present invention can be prepared as powder, granules, sustained-release granules, enteric-coated granules, solutions, eye drops, ellipsis, emulsions, suspensions, spirits, troches, perfumes, and limonadese according to conventional methods.
  • 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, gum acacia, alginate, gelatin, and calcium. These include phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
  • diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
  • Additives to tablets, powders, granules, capsules, pills, and troches according to the present invention include corn starch, potato starch, wheat starch, lactose, white sugar, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, and phosphoric acid.
  • Excipients such as cellulose (HPMC), HPMC 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethyl cellulose, sodium methyl cellulose, methyl cellulose, microcrystalline cellulose, dextrin.
  • binders can be used, Hydroxypropyl methyl cellulose, corn starch, agar powder, methyl cellulose, bentonite, hydroxypropyl starch, sodium carboxymethyl cellulose, sodium alginate, calcium carboxymethyl cellulose, calcium citrate, sodium lauryl sulfate, silicic anhydride, 1-hydroxy Propylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, Disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose
  • soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, Macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acids, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, Lubricants such as starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and light anhydrous silicic acid may be used.
  • Additives to the liquid according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.
  • a solution of white sugar, other sugars, or sweeteners, etc. may be used in the syrup according to the present invention, and if necessary, flavoring agents, colorants, preservatives, stabilizers, suspending agents, emulsifiers, thickening agents, etc. may be used.
  • Purified water can be used in the emulsion according to the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. can be used as needed.
  • Suspensions according to the present invention include acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, etc. Topics may be used, and surfactants, preservatives, stabilizers, colorants, and fragrances may be used as needed.
  • Injections according to the present invention include distilled water for injection, 0.9% sodium chloride injection, IV solution, dextrose injection, dextrose + sodium chloride injection, PEG, lactated IV solution, ethanol, propylene glycol, non-volatile oil - sesame oil.
  • solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristic acid, and benzene benzoate;
  • Solubilizing agents such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, Tween, nicotinic acid amide, hexamine, and dimethylacetamide;
  • 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 buffering agents such as gums
  • Isotonic agents such as sodium chloride
  • Stabilizers such as sodium bisulfite (NaHSO3) carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and
  • Suppositories according to the present invention include cacao oil, lanolin, witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, lecithin, Lanet wax, glycerol monostearate, Tween or Span, Imhausen, monolene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydrocote SP, S-70-XXA, S-70-XX75(S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Massaupol, Masupol-15, Neosupostal-
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the extract with at least one excipient, such as starch, calcium carbonate, and sucrose. ) or prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate talc are also used.
  • Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups.
  • various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included.
  • Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories.
  • Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.
  • composition according to the present invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat the disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, activity of the drug, and the type of patient's disease. It can be determined based on factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, 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 multiple times. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art to which the present invention pertains.
  • the pharmaceutical composition of the present invention can be administered to an individual through various routes. All modes of administration are contemplated, including oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, intrarectal injection, vaginal injection. It can be administered by internal insertion, ocular administration, otic administration, nasal administration, inhalation, spraying through the mouth or nose, dermal administration, transdermal administration, etc.
  • the pharmaceutical composition of the present invention is determined depending on the type of drug as the active ingredient along with various related factors such as the disease to be treated, the route of administration, the patient's age, gender, weight, and severity of the disease.
  • the present invention provides a method for preventing, improving, or treating fatty liver disease or obesity, comprising administering a hawthorn root extract or a composition containing the same as an active ingredient to an individual in need thereof.
  • the present invention provides the use of hawthorn root extract or a composition containing the same as an active ingredient to prevent, improve, or treat fatty liver disease or obesity.
  • the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient for preparing a preparation for preventing, improving or treating fatty liver disease or obesity.
  • “individual” refers to a subject in need of treatment for a disease, and more specifically, human or non-human primates, mice, rats, dogs, cats, horses, cows, etc. It may be a mammal, but is not limited thereto.
  • “administration” means providing a given composition of the present invention to an individual by any suitable method.
  • prevention refers to any action that suppresses or delays the onset of the desired disease
  • treatment refers to the improvement or improvement of the desired disease and its associated metabolic abnormalities by administration of the pharmaceutical composition according to the present invention. It refers to all actions that are beneficially changed, and “improvement” refers to all actions that reduce parameters related to the target disease, such as the degree of symptoms, by administering the composition according to the present invention.
  • the present invention provides a food composition for preventing or improving fatty liver disease or obesity, comprising hawthorn root extract as an active ingredient.
  • hawthorn, hawthorn root extract, solvent, fatty liver, composition, and transcription factor are directly applied to the food composition for preventing or improving fatty liver or obesity of the present application.
  • the food composition may be a health functional food composition, but is not limited thereto.
  • food includes functional foods and health functional foods.
  • the hawthorn root extract of the present invention When using the hawthorn root extract of the present invention as a food additive, the hawthorn root extract can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods.
  • the mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). In general, when producing food or beverages, the hawthorn root extract of the present invention may be added in an amount of 15% by weight or less, or 10% by weight 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 may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.
  • foods to which the above substances can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, These include alcoholic beverages and vitamin complexes, and include all health functional foods in the conventional sense.
  • the health drink composition according to the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, like conventional drinks.
  • the above-mentioned natural carbohydrates include 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.
  • a sweetener natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used.
  • the proportion of natural carbohydrates 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.
  • the composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, It may contain carbonating agents used in carbonated drinks. Additionally, the composition of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients can be used independently or in combination. The ratio of these additives is not very 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.
  • the present invention provides a cosmetic composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  • the hawthorn root extract of the present invention inhibits the accumulation of neutral fat and can be added to a cosmetic composition that has a cellulite reducing effect.
  • the hawthorn root extract of the present invention can be added as is or used together with other cosmetic ingredients, and can be used appropriately according to conventional methods.
  • the mixing amount of the active ingredient can be appropriately determined depending on the purpose of use.
  • the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier ingredients. etc. can be used.
  • the formulation of the cosmetic composition of the present invention is a powder or spray
  • lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient.
  • chlorofluorohydride may be additionally used. It may contain propellants such as carbon, propane-butane or dimethyl ether.
  • a solvent, solvating agent, or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, and propylene.
  • a carrier component such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, and propylene.
  • glycols 1,3-butylglycol oil, glycerol fatty esters, polyethylene glycol or fatty acid esters of sorbitan.
  • the carrier component includes water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
  • a liquid diluent such as ethanol or propylene glycol
  • a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester
  • Microcrystalline cellulose aluminum metahydroxide, bentonite, agar, or tracant may be used.
  • the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyl taurate, and sarcosinate.
  • fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linoline derivative, or ethoxylated glycerol fatty acid ester can be used.
  • the cosmetic composition of the present invention may further contain excipients including fluorescent substances, fungicides, hydrotropes-inducing substances, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, etc. .
  • the present invention provides a quasi-drug composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  • the term "quasi-drug” refers to fibers, rubber products, or similar products used for the purpose of treating, alleviating, treating, or preventing diseases in humans or animals. They have a weak effect on the human body or do not directly act on the human body, and refer to devices or Items that are non-machine or similar, or products that are used for sterilization, insecticide and similar purposes to prevent infectious diseases, and are used for the purpose of diagnosing, treating, mitigating, treating or preventing diseases in humans or animals. It refers to articles that are not instruments, machines, or devices among articles used for the purpose of having a pharmacological effect on the structure and function of humans or animals, excluding articles that are not instruments, machines, or devices.
  • the hawthorn root extract can be added as is or used together with other quasi-drug ingredients, and can be used appropriately according to conventional methods.
  • the mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment).
  • the quasi-drug composition of the present invention may be a disinfectant cleaner, toothpaste, shower foam, gargle, wet tissue, detergent, soap, hand wash, humidifier filler, mask, ointment, or filter filler.
  • the present invention provides a composition for inhibiting adipocyte differentiation in vitro, comprising hawthorn root extract as an active ingredient.
  • Hawthorn ( Crataegus pinnatifida ) root extract was obtained from the Korea Plant Extract Bank (Ochang-eup, Chungcheongbuk-do). The extract was extracted with 100% methanol, and after extraction, the extract was filtered and vacuum evaporated to dryness. To be used in test tube experiments, the obtained extract was dissolved in dimethyl sulfoxide (DMSO) at 50 mg/ml and then diluted. For preclinical experiments, the obtained extract was dissolved in sterile water and used.
  • DMSO dimethyl sulfoxide
  • Inguinal white adipose tissue ('iWAT'), epididymal white adipose tissue ('eWAT'), and liver were extracted from mice and then placed in 4% paraformaldehyde. After storing for 1 day and fixing the tissue, the tissue was embedded in paraffin, cut into 5 ⁇ m thick sections, stained with H & E according to standard protocols, and then examined under a microscope.
  • AST Aspartate aminotransferase
  • ALT alanine aminotransferase
  • 3T3-L1 preadipocytes were seeded in 6-well plates, and after 8 days of differentiation, they were analyzed using a fluorescence microscope and flow cytometer using the DCFDA/H2DCFDA-Celluar ROS Assay Kit (Abcam, US), referring to the supplier's protocol. .
  • DPPH 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity
  • CRE was diluted with DMSO to 2, 4, 6, 8, and 10 ⁇ g/ml, then vortexed, and 10.0039 g of DPPH was diluted in ethanol to obtain DPPH.
  • Reagents were prepared. Afterwards, ascorbic acid was used as a standard material, and CRE or ascorbic acid vortexed into the reagent was injected at the above concentration and reacted in the dark for 30 minutes. Afterwards, the absorbance was measured at 517 nm three times.
  • CRE was diluted with DMSO to 2, 4, 6, 8, and 10 ⁇ g/ml, then vortexed, and ABTS was added with potassium persulfate. (potassium persulfate) was added, dissolved in distilled water, and left at room temperature for 12 hours to prepare the ABTS reagent. Afterwards, CRE or ascorbic acid vortexed using ascorbic acid as a standard substance was injected into the ABTS reagent according to the above concentration and reacted in the dark for 30 minutes. Afterwards, the absorbance was measured three times at 734 nm.
  • AST Aspartate aminotransferase
  • ALT Alanine aminotransferase
  • ALP Alkaline phosphatase
  • TP Total protein
  • ALB Albumin
  • T-BIL total bilirubin
  • D-BIL Direct bilirubin
  • BUN Blood in plasma Urea nitrogen (CREA), creatinine (CREA), triglyceride (TG), and cholesterol (CHO) levels were measured using an Olympus AU 400 (Olympus Optical, Tokyo, Japan) analyzer according to the manufacturer's procedures.
  • 3T3-L1 preadipocytes were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% bovine calf serum (BCS). 3T3-L1 preadipocytes were seeded in a 6-well plate when confluence reached 80% in a 100-pi experimental dish and grown to reach confluence for 2 days. At 2 days after confluence, to initiate adipocyte differentiation, medium was supplemented with 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.25 mM dexamethasone (DEX), 1 ⁇ g/mL insulin, 0.125 mM India. It was changed to Differentiation medium I (DMI) containing metacin. Afterwards, maturation of adipocytes was induced for 6 days with differentiation medium II (DMII), and differentiation was performed for a total of 8 days.
  • DMEM Dulbecco's modified Eagle's medium
  • BCS bovine calf serum
  • 3T3-L1 preadipocytes were induced to differentiate in the presence or absence of CRE.
  • Adipocytes on day 8 of differentiation were washed with phosphate-buffered saline (PBS), fixed with paraformaldehyde, and stained with filtered 0.3% Oil Red O solution for 30 minutes at room temperature.
  • the reaction product was then dissolved in isopropyl alcohol. The absorbance of the reaction was measured at 450 nm.
  • 3T3-L1 preadipocytes were seeded in 96-well plates. After culturing, cells were treated with CRE or DMSO for 48 hours. MTT (3-(4,5)-dimethylthiazo-2-yl-2,5-diphenyltetrazolium bromide) solution was added to each well and incubated at 37°C for 4 hours. The reaction product was dissolved in isopropyl alcohol. The absorbance of the reaction was measured at 595 nm.
  • 3T3-L1 preadipocytes were induced to differentiate in the presence or absence of CRE.
  • Adipocytes on day 8 of differentiation were washed with phosphate-buffered saline (PBS), and the cells were dissolved in 5% NP-40 lysis buffer. Afterwards, it was measured using a Triglyceride assay kit according to the manufacturer's procedure.
  • PBS phosphate-buffered saline
  • 3T3-L1 preadipocytes on day 8 of differentiation were lysed with radioimmunoprecipitation assay (RIPA) lysis buffer containing phosphatase inhibitors and protease inhibitors.
  • Total protein (30 ⁇ g) content was separated on a 10% SDS-polyacrylamide gel by electrophoresis and transferred to a nitrocellulose membrane.
  • Membranes were blocked with 5% skim milk in PBS for 1 hour, incubated with relevant primary antibodies overnight at 4°C, and then membranes were washed three times for 10 minutes with TBS-T buffer. The membrane was then reacted with HRP-conjugated secondary antibody for 1 hour. Protein bands were visualized using ECL, detected on a Fusion Solo detector (Vilber Lourmat, Marne La Vallee, France), and evaluated by normalizing to ⁇ -actin.
  • RIPA radioimmunoprecipitation assay
  • 3T3-L1 preadipocytes were seeded in a 6-well plate and differentiated into groups according to the presence or absence of CRE.
  • 3T3-L1 cells on day 8 of differentiation were washed with PBS, and then total RNA was extracted from the cells using Trizol reagent.
  • the cDNA library was synthesized using the PrimeScript TM RT reagent kit (TaKaRa Bio). mRNA expression levels were quantified by analysis of cDNA implemented by the iCycler iQTM Real-Time PCR Detection System (Bio-Rad Laboratories, USA) using SYBR Green (TOYOBO, Japan), normalized to ⁇ -actin, and relative Levels of gene expression were expressed as folds.
  • Example 1 Confirmation of the inhibitory effect of hawthorn root extract (CRE) on adipocyte differentiation in 3T3-L1 preadipocytes.
  • CRE hawthorn root extract
  • adiponectin, C/EBP ⁇ , and PPAR ⁇ which are genes involved in adipocyte differentiation when treated with CRE. It was confirmed that the mRNA expression of SREBP-1 decreased in a concentration-dependent manner, and as shown in Figure 1e, the protein expression of C/EBP ⁇ and PPAR ⁇ also decreased when CRE was treated.
  • ROS reactive oxygen species
  • the radical scavenging ability of CRE was evaluated through IC50 and was found to be 23.674 ⁇ 1.452 ⁇ g/mL in the ABTS assay and 24.618 ⁇ 0.254 ⁇ g/mL in the DPPH assay, which is similar to that of Ascorbic acid, which is the control group.
  • the value was about 4 times higher and that the radical scavenging ability was very high at 1/4 of that of ascorbic acid.
  • hawthorn root extract effectively inhibits adipocyte differentiation without cytotoxicity, has an antioxidant effect, reduces lipid accumulation in adipocytes, and participates in adipocyte differentiation and maturation. It was confirmed that it suppresses the expression of transcription factors or adiponectin, and it was confirmed that it inhibited liver steatosis and improved liver damage or suppressed fat production caused by a high-fat diet in mice fed a high-fat diet. These results showed that hawthorn root extract was used to treat fatty liver or liver disease. It shows that it can be used as a drug and health functional food for preventing, treating, or improving obesity, and as a substance for their development.
  • hawthorn root extract effectively inhibits adipocyte differentiation without cytotoxicity, reduces lipid accumulation in adipocytes, and inhibits the expression of transcription factors involved in adipocyte differentiation and maturation.
  • the effect of inhibiting hepatic steatosis and improving liver damage caused by a high-fat diet was confirmed in high-fat diet mice, and the inhibition of fat production was confirmed in high-fat diet mice. Therefore, the hawthorn root extract of the present invention is expected to be useful in pharmaceutical compositions and health functional foods for preventing, treating, or improving fatty liver disease or obesity, and has industrial applicability.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Botany (AREA)
  • Biotechnology (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Microbiology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Dermatology (AREA)
  • Child & Adolescent Psychology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medical Informatics (AREA)
  • Diabetes (AREA)
  • Obesity (AREA)
  • Hematology (AREA)
  • Birds (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

The present invention relates to a composition for preventing, treating or alleviating fatty liver or obesity, comprising an extract of Crataegus pinnatifida Bunge roots. The present inventors identified that the extract of Crataegus pinnatifida Bunge roots effectively inhibits preadipocyte cell differentiation without cytotoxicity, reduces lipid accumulation in adipocytes, and inhibits the expression of a transcription factor or adiponectin, which is involved in the formation and differentiation of adipocytes. In addition, an adiposis inhibitory effect and the effect of alleviating liver damage caused by high-fat feeding has been identified in high-fat diet mice, and it has been identified that adipogenesis and lipid accumulation are inhibited in high-fat diet mice. Therefore, Crataegus pinnatifida Bunge roots of the present invention can be effectively used as a pharmaceutical composition, a health functional food and the like for preventing, treating or alleviating fatty liver of obesity.

Description

산사나무 뿌리 추출물을 유효성분으로 포함하는 지방간 또는 비만의 예방 또는 치료용 약학적 조성물Pharmaceutical composition for preventing or treating fatty liver disease or obesity containing hawthorn root extract as an active ingredient
본 발명은 산사나무(Crataegus pinnatifida) 추출물을 유효성분으로 포함하는 지방간 또는 비만의 예방, 개선 또는 치료용 조성물 등에 관한 것이다.The present invention relates to a composition for preventing, improving or treating fatty liver disease or obesity, including a hawthorn ( Crataegus pinnatifida ) extract as an active ingredient.
본 발명은 2022년 12월 1일에 출원된 한국특허출원 제10-2022-0165566호 및 2022년 12월 1일에 출원된 한국특허출원 제10-2022-0165578호에 기초한 우선권을 주장하며, 해당 출원의 명세서 및 도면에 개시된 모든 내용은 본 출원에 원용된다. The present invention claims priority based on Korean Patent Application No. 10-2022-0165566 filed on December 1, 2022 and Korean Patent Application No. 10-2022-0165578 filed on December 1, 2022. All contents disclosed in the specification and drawings of the application are incorporated into this application.
비만은 일반적으로 체내에 지방조직이 과다한 상태인 것을 의미하며, 음식물로 섭취한 에너지가 신체활동 등으로 소비한 에너지와 균형을 이루지 못하여 잉여의 에너지가 체지방으로 축적되는 현상이다. 비만은 유전적인 요인, 불규칙한 식습관이나 운동부족 또는 서구화되는 식생활에 의한 환경적인 영향, 우울감, 지루감, 과다한 스트레스에 의한 심리적인 영향, 갑상선이나 부신피질 호르몬 변화에 의한 병적인 요인 등 매우 다양한 원인에 의해 유발되는 것으로 알려져 있다. 최근에는 경제성장과 생활방식의 변화에 따라 식습관에도 많은 변화가 생겨 바쁜 현대인들은 패스트푸드 등의 고열량 식이와 적은 운동량으로 인하여 과체중 및 비만이 증가하고 있는 추세이다. Obesity generally refers to the state of excessive fatty tissue in the body, and is a phenomenon in which the energy consumed through food is not balanced with the energy consumed through physical activity, and the excess energy is accumulated as body fat. Obesity has a wide variety of causes, including genetic factors, environmental influences due to irregular eating habits, lack of exercise or westernized eating habits, depression, boredom, psychological effects due to excessive stress, and pathological factors due to changes in thyroid or adrenal cortex hormones. It is known to be caused by Recently, with economic growth and changes in lifestyle, there have been many changes in eating habits, and busy modern people are increasingly overweight and obese due to high-calorie diets such as fast food and little exercise.
또한, 비만은 지방세포의 비대에 의해 유발되므로, 지방세포 분화의 억제는 비만 치료에 도움이 될 수 있다. 종래에 지방세포 분화 및 지질 축적의 억제와 관련된 항비만 연구가 보고된 바 있으며, 이 때 3T3-L1 마우스 섬유아세포는 지방 전구세포의 분화를 위한 잘 특성화된 모델로 일반적으로 시험관 내 연구에 사용된다. Additionally, since obesity is caused by hypertrophy of adipocytes, inhibition of adipocyte differentiation may be helpful in treating obesity. Previously, anti-obesity studies related to the inhibition of adipocyte differentiation and lipid accumulation have been reported, and 3T3-L1 mouse fibroblasts are a well-characterized model for the differentiation of preadipocytes and are generally used for in vitro studies. .
한편, 간은 우리 몸에서 대사, 해독 및 배설과 관련하여 가장 중요한 장기이다. 바이러스, 음주, 약물, 영양불량, 비만 등에 의해 간 기능이 저하되는데, 특히 현대인의 고당, 고지방 섭취에 따른 지방간은 나아가 지방간염으로 진행될 수 있다. Meanwhile, the liver is the most important organ in our body related to metabolism, detoxification, and excretion. Liver function deteriorates due to viruses, alcohol, drugs, malnutrition, obesity, etc. In particular, fatty liver caused by high sugar and high fat intake in modern people can further progress to steatohepatitis.
지방간은 알코올성 지방간과 비알코올성 지방간으로 나누어지는데, 알코올에 기인하지 않은 지방축적으로 5% 이상 무게가 증가하면 비알코올성 지방간 질환으로 정의한다. 음주와는 무관한 비알코올성 지방간질환(nonalcoholic fatty liver disease)은 전 세계적으로 적게는 6.3%, 많게는 33%가 보고되고 있으며, 적지만 정상체중 성인에게도 비알코올성 지방간병변이 나타나는 것으로 보고되고 있다. 비알코올성 지방간질환은 지방조직으로부터 간으로 유리지방산 흡수가 증가하거나, 간에서 중성지방의 생합성이 증가하여 발생하는데, 특히 인슐린 저항성이 일어나는 경우 지방조직에서 지방분해가 촉진되어 간으로 유입되는 지방산을 증가시키고, 근육세포에서 포도당 이용 장애를 유발하여 고인슐린혈증 및 탄수화물의 지방 전환(de novo lipogenesis)을 증가시켜 간내 지방축적을 더욱 증가시킨다.Fatty liver is divided into alcoholic fatty liver disease and non-alcoholic fatty liver disease. If the weight increases by more than 5% due to fat accumulation not caused by alcohol, it is defined as non-alcoholic fatty liver disease. Nonalcoholic fatty liver disease, which is unrelated to drinking, is reported in as few as 6.3% and as many as 33% worldwide, and non-alcoholic fatty liver lesions are reported to occur even in a small number of normal-weight adults. Non-alcoholic fatty liver disease is caused by increased absorption of free fatty acids from adipose tissue into the liver or increased biosynthesis of neutral fat in the liver. In particular, when insulin resistance occurs, lipolysis is promoted in adipose tissue, increasing fatty acids flowing into the liver. It causes impaired glucose utilization in muscle cells, increasing hyperinsulinemia and carbohydrate conversion to fat (de novo lipogenesis), further increasing fat accumulation in the liver.
비만으로 인한 비알코올성 지방간은 체내의 중성지방이 축적되면서 단순 지방간(steatosis)부터, 증상이 심하면 지방간염(non-alcoholic steatohepatitis)으로 진행되며, 다른 질병과는 달리 일반적 증상 없이 간경변, 간 기능 상실, 간암 등의 복잡하고 점진적 간 손상을 진행시키기 때문에 조기 치료 및 예방이 필요하다. Non-alcoholic fatty liver disease caused by obesity progresses from simple fatty liver (steatosis) to non-alcoholic steatohepatitis (non-alcoholic steatohepatitis) in severe cases, as neutral fat accumulates in the body. Unlike other diseases, it progresses to cirrhosis, loss of liver function, etc. without general symptoms. Early treatment and prevention are necessary because it causes complex and progressive liver damage such as liver cancer.
비만은 식이요법, 규칙적인 운동과 더불어 행동요법과 같은 생활습관의 개선, 및 식욕 억제제와 지방 흡수 억제제와 같은 약물을 통해 치료할 수 있다. 비만은 만성 질환이기 때문에 약물치료를 시도하는 경우 장기간의 사용이 필요하며, 현재 국내에서 3개월 이상 장기간 사용이 허가된 제품으로는 식욕억제제인 시부트라민(sibutramine)과 지방분해효소 억제제인 올리스타트(orlistat)가 있다. 그러나 이러한 비만 치료 약물들은 대부분 중추신경계에 작용하여 식욕을 조절하는 향정신성의약품들이므로 두통 및 구토 등의 부작용을 동반하며 남용 우려 등의 문제점이 있다. 따라서 상기 시판중인 항비만제의 부작용을 해결할 수 있는 안정성이 높고 항비만 효과가 우수한 소재를 개발하기 위한 연구가 활발히 이루어지고 있다.Obesity can be treated through diet, regular exercise, lifestyle changes such as behavioral therapy, and medications such as appetite suppressants and fat absorption inhibitors. Since obesity is a chronic disease, attempting drug treatment requires long-term use. Currently, products approved for long-term use in Korea for more than 3 months include the appetite suppressant sibutramine and the lipolytic enzyme inhibitor orlistat. ). However, most of these obesity treatment drugs are psychotropic drugs that act on the central nervous system to control appetite, so they come with side effects such as headaches and vomiting and have problems such as concerns about abuse. Therefore, research is being actively conducted to develop materials with high stability and excellent anti-obesity effect that can solve the side effects of the commercially available anti-obesity drugs.
산사나무(Crataegus pinnatifida)는 장미목 장미과에 해당하는 낙엽 교목으로 국내에서는 전국 산지에서 자생하며 열매의 경우 예로부터 식용 및 약용으로 즐겨 사용되었으나, 산사나무 뿌리에 대해서는 생리활성 효과가 많이 알려진 바가 없다. Hawthorn (Crataegus pinnatifida) is a deciduous tree belonging to the Rosaceae family of the rose order. In Korea, it grows wild in mountainous regions across the country. Its fruits have been used for food and medicinal purposes since ancient times, but not much is known about the physiological effects of the roots of hawthorn.
이에, 본 발명자들은 산사나무 뿌리 추출물의 지방간 개선 효과 또는 항비만 활성을 확인하여 본 발명을 완성하게 되었다.Accordingly, the present inventors completed the present invention by confirming the fatty liver improvement effect or anti-obesity activity of the hawthorn root extract.
이에 본 발명자들은 종래 항비만제의 문제점을 해결할 수 있는 안정성이 높고 항지방간 또는 항비만 효과가 우수한 지방간 또는 비만의 예방 또는 치료용 조성물을 개발하고자 노력한 결과, 본 발명의 산사나무 뿌리 추출물이 뛰어난 항지방간 또는 항비만 효과를 가지는 것을 확인하고 본 발명을 완성하였다.Accordingly, the present inventors have made efforts to develop a composition for preventing or treating fatty liver or obesity with high stability that can solve the problems of conventional anti-obesity drugs and excellent anti-fatty liver or anti-obesity effects. As a result, the hawthorn root extract of the present invention has been found to be an excellent anti-obesity drug. The present invention was completed after confirming that it had an anti-fatty liver or anti-obesity effect.
따라서, 본 발명의 목적은 산사나무 뿌리 추출물을 유효성분으로 포함하는 지방간의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.Accordingly, the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating fatty liver comprising hawthorn root extract as an active ingredient.
본 발명의 다른 목적은 산사나무 뿌리 추출물을 유효성분으로 포함하는 지방간의 예방 또는 개선용 식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a food composition for preventing or improving fatty liver disease containing hawthorn root extract as an active ingredient.
본 발명의 목적은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating obesity containing hawthorn root extract as an active ingredient.
본 발명의 다른 목적은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 식품 조성물을 제공하는 것이며, 또 다른 목적은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 화장료 조성물을 제공하는 것이다.Another object of the present invention is to provide a food composition for preventing or improving obesity containing hawthorn root extract as an active ingredient, and another object is to provide a cosmetic composition for preventing or improving obesity containing hawthorn root extract as an active ingredient. A composition is provided.
본 발명의 또 다른 목적은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 의약외품 조성물을 제공하는 것이다.Another object of the present invention is to provide a quasi-drug composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.
상기 본 발명의 목적을 달성하기 위하여, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 지방간의 예방 또는 치료용 약학적 조성물을 제공한다.In order to achieve the object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating fatty liver comprising hawthorn root extract as an active ingredient.
또한, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 지방간의 예방 또는 개선용 식품 조성물을 제공한다.Additionally, the present invention provides a food composition for preventing or improving fatty liver disease containing hawthorn root extract as an active ingredient.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물을 이를 필요로 하는 개체에 투여하는 단계를 포함하는, 지방간의 예방, 개선, 또는 치료 방법을 제공한다.Additionally, the present invention provides a method for preventing, improving, or treating fatty liver disease, comprising administering a hawthorn root extract or a composition containing the same as an active ingredient to an individual in need thereof.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 지방간 예방, 개선, 또는 치료 용도를 제공한다.In addition, the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient to prevent, improve, or treat fatty liver.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 지방간 예방, 개선 또는 치료 제제를 제조하기 위한 용도를 제공한다.In addition, the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient for preparing a preparation for preventing, improving, or treating fatty liver disease.
또한, 본 발명은 지방간의 예방, 개선, 또는 치료용 약제를 제조하기 위한 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 용도를 제공한다.In addition, the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient for preparing a medicine for preventing, improving, or treating fatty liver disease.
본 발명의 일실시예에서, 상기 산사나무 뿌리 추출물은 물, 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane), 석유에테르(petroleum ether), 디클로로메탄(dichloromethane), 아임계 유체, 및 초임계 유체로 이루어진 군으로부터 선택된 하나 이상의 용매에 의한 추출물일 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the hawthorn root extract contains water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, and ethyl acetate. one or more solvents selected from the group consisting of acetate, methylene chloride, hexane, cyclohexane, petroleum ether, dichloromethane, subcritical fluids, and supercritical fluids It may be an extract, but is not limited thereto.
본 발명의 일실시예에서, 상기 지방간은 비알콜성 지방간일 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the fatty liver may be non-alcoholic fatty liver, but is not limited thereto.
본 발명의 일실시예에서, 상기 조성물은 고지방식이로 유도된 간 손상을 개선할 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the composition can improve liver damage induced by a high-fat diet, but is not limited thereto.
본 발명의 일실시예에서, 상기 조성물은 혈청 ALT 또는 AST 활성을 감소시킬 수 있고, 바람직하게는 상기 조성물은 고지방식이군의 혈청 ALT 또는 AST 활성을 감소시킬 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the composition can reduce serum ALT or AST activity, and preferably, the composition can reduce serum ALT or AST activity in a high-fat diet group, but is not limited thereto.
본 발명의 일실시예에서, 상기 산사나무 뿌리 추출물은 지방세포 분화 및 성숙에 관여하는 전사인자 또는 아디포넥틴의 발현을 억제할 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the hawthorn root extract may inhibit the expression of transcription factors or adiponectin involved in adipocyte differentiation and maturation, but is not limited thereto.
본 발명의 일실시예에서, 상기 전사인자는 퍼옥시좀 증식제 활성화 수용체 감마(PPARγ), 스테롤 조절인자 결합 단백질-1(SREBP-1) 또는 CCAAT-인핸서 결합 단백질 알파(C/EBPα)일 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the invention, the transcription factor may be peroxisome proliferator activated receptor gamma (PPARγ), sterol regulator binding protein-1 (SREBP-1), or CCAAT-enhancer binding protein alpha (C/EBPα). However, it is not limited to this.
본 발명의 일실시예에서, 상기 식품 조성물은 건강기능성 식품 조성물일 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the food composition may be a health functional food composition, but is not limited thereto.
본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 치료용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating obesity containing hawthorn root extract as an active ingredient.
또한, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 식품 조성물을 제공한다.Additionally, the present invention provides a food composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
또한, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 화장료 조성물을 제공한다.Additionally, the present invention provides a cosmetic composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
또한, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 의약외품 조성물을 제공한다.In addition, the present invention provides a quasi-drug composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물을 이를 필요로 하는 개체에 투여하는 단계를 포함하는, 비만의 예방, 개선, 또는 치료 방법을 제공한다.Additionally, the present invention provides a method for preventing, improving, or treating obesity, comprising administering a hawthorn root extract or a composition containing the same as an active ingredient to an individual in need thereof.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 비만의 예방, 개선, 또는 치료 용도를 제공한다.Additionally, the present invention provides a use of hawthorn root extract or a composition containing it as an active ingredient to prevent, improve, or treat obesity.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 비만의 예방, 개선 또는 치료 제제를 제조하기 위한 용도를 제공한다.In addition, the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient for preparing a preparation for preventing, improving or treating obesity.
또한, 본 발명은 비만의 예방, 개선, 또는 치료용 약제를 제조하기 위한 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 용도를 제공한다.Additionally, the present invention provides the use of a hawthorn root extract or a composition containing it as an active ingredient for preparing a medicine for preventing, improving, or treating obesity.
본 발명의 일실시예에서, 상기 산사나무 뿌리 추출물은 물, 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane), 석유에테르(petroleum ether), 디클로로메탄(dichloromethane), 아임계 유체, 및 초임계 유체로 이루어진 군으로부터 선택된 하나 이상의 용매에 의한 추출물일 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the hawthorn root extract contains water, alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, and ethyl acetate. one or more solvents selected from the group consisting of acetate, methylene chloride, hexane, cyclohexane, petroleum ether, dichloromethane, subcritical fluids, and supercritical fluids It may be an extract, but is not limited thereto.
본 발명의 일실시예에서, 상기 산사나무 뿌리 추출물은 하기 성질중 하나 이상을 가질 수 있으나, 이에 제한되는 것은 아니다:In one embodiment of the present invention, the hawthorn root extract may have one or more of the following properties, but is not limited thereto:
i) 지방세포의 분화를 억제;i) Inhibit differentiation of adipocytes;
ii) 세포 내 지질 축적을 감소; 및ii) reduce intracellular lipid accumulation; and
iii) 지방 세포 분화중 유도되는 산화 스트레스를 감소.iii) Reduces oxidative stress induced during adipocyte differentiation.
본 발명의 일실시예에서, 상기 산사나무 뿌리 추출물은 지방세포 분화 및 성숙에 관여하는 전사인자 또는 아디포넥틴의 발현을 억제할 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the hawthorn root extract may inhibit the expression of transcription factors or adiponectin involved in adipocyte differentiation and maturation, but is not limited thereto.
본 발명의 일실시예에서, 상기 전사인자는 퍼옥시좀 증식제 활성화 수용체 감마(PPARγ), 스테롤 조절인자 결합 단백질-1(SREBP-1) 또는 CCAAT-인핸서 결합 단백질 알파(C/EBPα)일 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the invention, the transcription factor may be peroxisome proliferator activated receptor gamma (PPARγ), sterol regulator binding protein-1 (SREBP-1), or CCAAT-enhancer binding protein alpha (C/EBPα). However, it is not limited to this.
본 발명의 일실시예에서, 상기 식품 조성물은 건강기능성 식품 조성물일 수 있으나, 이에 제한되는 것은 아니다.In one embodiment of the present invention, the food composition may be a health functional food composition, but is not limited thereto.
본 발명자들은 산사나무 뿌리 추출물이 세포 독성 없이 지방 전구 세포 분화를 효과적으로 억제하고, 지방세포 내 지질 축적을 감소시킬 뿐만 아니라, 지방 세포 분화 및 성숙에 관여하는 전사인자의 발현을 억제함을 확인하였다. 또한 고지방 식이 마우스에서 고지방 식이로 인한 간 지방증의 저해 및 간 손상의 개선효과를 확인하였고, 고지방 식이 마우스에서 지방 생성의 억제를 확인하였다. 따라서 본 발명의 산사나무 뿌리 추출물은 지방간 또는 비만의 예방, 치료, 또는 개선용 약학적 조성물 및 건강기능식품 등으로 유용하게 사용될 수 있다.The present inventors confirmed that hawthorn root extract effectively inhibits adipocyte differentiation without cytotoxicity, reduces lipid accumulation in adipocytes, and inhibits the expression of transcription factors involved in adipocyte differentiation and maturation. In addition, the effect of inhibiting hepatic steatosis and improving liver damage caused by a high-fat diet was confirmed in high-fat diet mice, and the inhibition of fat production was confirmed in high-fat diet mice. Therefore, the hawthorn root extract of the present invention can be usefully used as a pharmaceutical composition and health functional food for preventing, treating, or improving fatty liver disease or obesity.
도 1a는 3T3-L1 지방 전구 세포를 595 nm 흡광도에서 MTT 분석으로 생존력을 측정한 결과를 나타낸 것이다. CON은 CRE와 동일한 양의 DMSO를 처리하였다.Figure 1a shows the results of measuring the viability of 3T3-L1 preadipocytes by MTT assay at 595 nm absorbance. CON was treated with the same amount of DMSO as CRE.
도 1b는 3T3-L1 지방 전구 세포를 Oil Red O 용액 염색한 후 관찰한 이미지(왼쪽), 및 Oil Red O(ORO) 용액의 함량을 450 nm 흡광도로 측정한 결과(오른쪽, *p <0.05)를 나타낸 것이다.Figure 1b shows an image observed after staining 3T3-L1 preadipocytes with Oil Red O solution (left), and the result of measuring the content of Oil Red O (ORO) solution at absorbance at 450 nm (right, *p <0.05). It represents.
도 1c는 3T3-L1 지방 전구 세포 내 트리글리세라이드 함량을 570 nm 흡광도로 측정한 결과를 나타낸 것이다. *p <0.05Figure 1c shows the results of measuring the triglyceride content in 3T3-L1 preadipocytes using absorbance at 570 nm. *p <0.05
도 1d는 3T3-L1 지방 전구 세포에서 Adiponectin, C/EBPα, PPARγ 및 SREBP-1의 mRNA 발현 수준을 측정한 결과이다. *p <0.05Figure 1d shows the results of measuring the mRNA expression levels of Adiponectin, C/EBPα, PPARγ, and SREBP-1 in 3T3-L1 preadipocytes. *p <0.05
도 1e는 3T3-L1 지방 전구 세포에서 C/EBPα 및 PPARγ의 단백질 발현 수준을 측정한 결과이다. *p <0.05Figure 1e shows the results of measuring the protein expression levels of C/EBPα and PPARγ in 3T3-L1 preadipocytes. *p <0.05
도 2a는 iWAT, eWAT 및 간 조직을 H&E로 염색하여 지방구를 관찰한 이미지이다.Figure 2a is an image showing fat globules in iWAT, eWAT, and liver tissue stained with H&E.
도 2b는 iWAT 조직에서 SREBP-1 및 PPARγ의 단백질 발현 수준을 측정한 결과이다. *p <0.05Figure 2b shows the results of measuring the protein expression levels of SREBP-1 and PPARγ in iWAT tissue. *p <0.05
도 2c는 각 마우스 그룹의 간 대표 이미지를 나타낸 것이다.Figure 2c shows representative images of the liver of each mouse group.
도 2d는 각 마우스 그룹의 혈장 내 AST 및 ALT 수준을 측정한 결과를 나타낸 것이다. *p <0.05Figure 2d shows the results of measuring the plasma levels of AST and ALT in each mouse group. *p <0.05
도 3a는 3T3-L1 지방 전구 세포내 활성 산소를 형광 현미경을 통해 관찰한 이미지이다.Figure 3a is an image of reactive oxygen species in 3T3-L1 preadipocytes observed through a fluorescence microscope.
도 3b는 3T3-L1 지방 전구 세포내 활성 산소를 유세포 분석기를 사용하여 정량적으로 측정한 결과를 나타낸 것이다.Figure 3b shows the results of quantitative measurement of reactive oxygen species in 3T3-L1 preadipocytes using a flow cytometer.
도 4a는 각 마우스 그룹의 사료 섭취량 비율을 나타낸 것이다. **p <0.05 Figure 4a shows the feed intake ratio of each mouse group. **p <0.05
도 4b는 각 그룹별 마우스의 체중을 일주일 간격으로 측정한 결과를 나타낸것이다. *p <0.05Figure 4b shows the results of measuring the body weight of mice in each group at weekly intervals. *p <0.05
도 4c는 각 그룹별 지방 조직 및 장기의 무게를 측정한 결과(상단, *p <0.05 및 **p <0.01) 이에 따른 체중대비 비만도를 측정한 결과(하단)를 나타낸 것이다.Figure 4c shows the results of measuring the weight of adipose tissue and organs in each group (top, *p <0.05 and **p <0.01) and the results of measuring obesity relative to body weight (bottom).
도 4d는 각 마우스 그룹의 TG 함량(좌측), 총 콜레스테롤 함량(우측)을 측정한 결과를 나타낸 것이다. *p <0.05Figure 4d shows the results of measuring the TG content (left) and total cholesterol content (right) of each mouse group. *p <0.05
도 4e 내지 도 4g는 각 마우스 그룹의 복부 장기(도 4e), iWAT(도 4f), eWAT(도 4g)의 대표 이미지를 나타낸 것이다.Figures 4E to 4G show representative images of abdominal organs (Figure 4E), iWAT (Figure 4F), and eWAT (Figure 4G) of each mouse group.
본 발명자들은 본 발명의 산사나무 뿌리 추출물이 뛰어난 지방간 개선 효과를 가지는 것을 확인하고 본 발명을 완성하였다.The present inventors confirmed that the hawthorn root extract of the present invention has an excellent fatty liver improvement effect and completed the present invention.
또한 본 발명자들은 안정성이 높고 항비만 효과가 우수한 비만 예방 또는 치료용 조성물을 개발하고자 노력한 결과, 본 발명의 산사나무 뿌리 추출물이 뛰어난 항비만 효과를 가지는 것을 확인하고 본 발명을 완성하였다.In addition, the present inventors tried to develop a composition for preventing or treating obesity with high stability and excellent anti-obesity effect, and as a result, they confirmed that the hawthorn root extract of the present invention had excellent anti-obesity effect and completed the present invention.
이하, 본 발명에 대해 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 지방간 또는 비만의 예방, 개선 또는 치료용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing, improving, or treating fatty liver disease or obesity, comprising hawthorn root extract as an active ingredient.
본 발명에서 사용되는 용어 "산사나무(Crataegus pinnatifida for. pinnatifida)"는 장미과 산사나무속에 속하는 낙엽성 소교목으로 국내에서는 전국 산지에서 자생하고 산사나무의 잎과 꽃, 열매, 껍질, 뿌리는 예로부터 식용 또는 약용으로 사용되었다.The term "hawthorn (Crataegus pinnatifida for. pinnatifida)" used in the present invention is a deciduous small tree belonging to the genus of Hawthorn in the Rosaceae family. In Korea, it grows naturally in mountainous areas across the country, and the leaves, flowers, fruits, bark, and roots of the hawthorn tree have been used since ancient times. It was used for food or medicinal purposes.
본 발명에서 "산사나무 뿌리 추출물"은 상기 산사나무 뿌리의 추출처리에 의하여 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 조정제물이나 정제물, 또는 이들의 혼합물 등 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함한다.In the present invention, “hawthorn root extract” refers to an extract obtained by extraction treatment of the hawthorn root, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a crude product or purified product of the extract, or a mixture thereof. This includes the extract itself and all formulations of extracts that can be formed using the extract.
본 발명의 산사나무 뿌리 추출물에 있어서, 상기 산사나무 뿌리를 추출하는 방법은 특별히 제한되지 아니하며, 당해 기술분야에서 통상적으로 사용하는 방법에 따라 추출할 수 있다. 상기 추출 방법의 비제한적인 예로는, 열수 추출법, 초음파 추출법, 여과법, 환류 추출법 등을 들 수 있으며, 이들은 단독으로 수행되거나 2종 이상의 방법을 병용하여 수행될 수 있으나, 이에 제한되지 않는다.In the hawthorn root extract of the present invention, the method of extracting the hawthorn root is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, and reflux extraction, which may be performed alone or in combination of two or more methods, but are not limited thereto.
본 발명에 있어서, 산사나무 뿌리 추출물은 물, 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane), 석유에테르(petroleum ether), 디클로로메탄(dichloromethane), 아임계 유체, 및 초임계 유체로 이루어진 군으로부터 선택된 하나 이상의 용매에 의한 추출물일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the hawthorn root extract is water, alcohol with 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, and methylene. Extraction with one or more solvents selected from the group consisting of methylene chloride, hexane, cyclohexane, petroleum ether, dichloromethane, subcritical fluid, and supercritical fluid. However, it is not limited to this.
본 발명에서 산사나무 뿌리를 추출하는 데에 사용되는 용매의 종류는 특별히 제한되지 아니하며, 천연물로부터 추출물을 추출하는 당업계에 공지된 통상적인 방법에 따라, 즉 통상적인 온도, 압력의 조건 하에서 통상적인 용매를 사용하여 추출할 수 있다. 예를 들어, 물, 탄소수 1 내지 6개의 유기용매 및 아임계 또는 초임계 유체로 이루어진 군에서 선택된 하나 이상의 용매로 추출될 수 있다. 상기 탄소수 1 내지 6개의 유기용매는 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 디클로로메탄(dichloromethane), 시클로헥산(cyclohexane) 및 석유에테르(petroleum ether)로 이루어진 군으로부터 선택된 하나 이상인 것일 수 있으나, 이에 제한되는 것은 아니다. 본 발명의 일 구현예에 따르면 상기 용매는 50% 내지 100%, 55% 내지 100%, 60% 내지 100%, 65% 내지 100%, 70% 내지 100%, 75% 내지 100%, 80% 내지 100%, 85% 내지 100%, 90% 내지 100%, 95% 내지 100%, 또는 100% 용매일 수 있으나, 이에 제한되는 것은 아니다. 본 발명의 다른 구현예에 따르면 상기 용매는 50% 내지 100%, 55% 내지 100%, 60% 내지 100%, 65% 내지 100%, 70% 내지 100%, 75% 내지 100%, 80% 내지 100%, 85% 내지 100%, 90% 내지 100%, 95% 내지 100%, 또는 100% 메탄올일 수 있으며, 바람하게는 100% 메탄올일 수 있으나, 이에 제한되는 것은 아니다. 추출방법은 상기 용매를 사용하여 냉침, 온침, 가열 등 통상의 추출방법이 사용 가능하다.The type of solvent used to extract hawthorn roots in the present invention is not particularly limited, and is performed according to a conventional method known in the art for extracting extracts from natural products, that is, under conventional conditions of temperature and pressure. It can be extracted using a solvent. For example, it may 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 solvent having 1 to 6 carbon atoms is alcohol, acetone, ether, benzene, chloroform, ethyl acetate, and methylene chloride. It may be one or more selected from the group consisting of chloride, hexane, dichloromethane, cyclohexane, and petroleum ether, but is not limited thereto. According to one embodiment of the present invention, the solvent is 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 80%. It may be 100%, 85% to 100%, 90% to 100%, 95% to 100%, or 100% solvent, but is not limited thereto. According to another embodiment of the present invention, the solvent is 50% to 100%, 55% to 100%, 60% to 100%, 65% to 100%, 70% to 100%, 75% to 100%, 80% to 80%. It may be 100%, 85% to 100%, 90% to 100%, 95% to 100%, or 100% methanol, preferably 100% methanol, but is not limited thereto. Conventional extraction methods such as cold immersion, warm immersion, and heating can be used using the above solvent.
본 발명에 있어서, 지방간은 비알콜성 지방간일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, fatty liver may be non-alcoholic fatty liver, but is not limited thereto.
본 발명에 있어서, 조성물은 고지방식이로 유도된 간 손상을 개선할 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the composition can improve liver damage induced by a high-fat diet, but is not limited thereto.
본 발명에 있어서, 조성물은 혈청 ALT 또는 AST 활성을 감소시킬 수 있고, 바람직하게는 상기 조성물은 고지방식이군의 혈청 ALT 또는 AST 활성을 감소시킬 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the composition can reduce serum ALT or AST activity, and preferably, the composition can reduce serum ALT or AST activity in the high-fat diet group, but is not limited thereto.
본 발명에 있어서, 산사나무 뿌리 추출물은 지방세포 분화 및 성숙에 관여하는 전사인자 또는 아디포넥틴의 발현을 억제할 수 있으나, 이에 제한되는 것은 아니다.In the present invention, hawthorn root extract can inhibit the expression of transcription factors or adiponectin involved in adipocyte differentiation and maturation, but is not limited thereto.
본 발명에 있어서, 전사인자는 퍼옥시좀 증식제 활성화 수용체 감마(PPARγ스테롤 조절인자 결합 단백질-1(SREBP-1) 또는 CCAAT-인핸서 결합 단백질 알파(C/EBPα)일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the transcription factor may be peroxisome proliferator-activated receptor gamma (PPARγ sterol regulator binding protein-1 (SREBP-1) or CCAAT-enhancer binding protein alpha (C/EBPα), but is limited thereto. no.
본 발명에 있어서, 산사나무 뿌리 추출물은 하기 성질중 하나 이상을 가질 수 있으나, 이에 제한되는 것은 아니다:In the present invention, the hawthorn root extract may have one or more of the following properties, but is not limited thereto:
i) 지방세포의 분화를 억제;i) Inhibit differentiation of adipocytes;
ii) 세포 내 지질 축적을 감소; 및ii) reduce intracellular lipid accumulation; and
iii) 지방 세포 분화중 유도되는 산화 스트레스를 감소.iii) Reduces oxidative stress induced during adipocyte differentiation.
본 발명의 일실시예에서는 산사나무 뿌리 추출물이 세포 독성을 야기하지 않고 3T3-L1 지방 전구 세포 분화를 억제하여 지방 함량을 감소시킴을 확인하였다 (실시예 1 참조).In one embodiment of the present invention, it was confirmed that hawthorn root extract reduces fat content by inhibiting 3T3-L1 preadipocyte differentiation without causing cytotoxicity (see Example 1).
본 발명의 다른 일실시예에서는, 산사나무 뿌리 추출물이 HFD 유도된 비만마우스에서 지방증의 저해 효과 및 간 손상의 개선 효과를 확인하였다 (실시예 2 참조).In another example of the present invention, hawthorn root extract was confirmed to have an inhibitory effect on steatosis and an improvement in liver damage in HFD-induced obese mice (see Example 2).
본 발명의 용어 "항산화"는 산화를 억제하는 작용을 의미하는 것으로, 인체는 산화촉진물질(prooxidant)과 산화억제물질(antioxidant)이 균형을 이루고 있으나 여러 가지 요인들에 의하여 이런 균형상태가 불균형을 이루게 되고 산화촉진 쪽으로 기울게 되면, 산화적 스트레스(oxidative stress)가 유발되어 잠재적인 세포손상 및 병리적 질환을 일으키게 된다. 이러한 산화적 스트레스의 직접적 원인이 되는 활성산소종(reactive oxygen species, ROS)은 불안정하고 반응성이 높아 여러 생체물질과 쉽게 반응하고, 체내 고분자들을 공격하여 세포와 조직에 비가역적인 손상을 일으키거나 돌연변이, 세포독성 및 발암 등을 초래하게 된다. NO, HNO2, ONOO-와 같은 활성질소종(reactive nitrogen species, RNS)은 염증 반응 시 대식세포 호중구 및 다른 면역 세포 들의 면역반응으로 인해 다량 생성되며, 이때 ROS도 같이 생성된다. 상기와 같은 활성산소는 체내에서 세포를 산화시켜 파괴시키며, 그에 따라 각종 질환에 노출되게 된다.The term "antioxidant" in the present invention refers to the action of suppressing oxidation. The human body has a balance of prooxidants and antioxidants, but this balance can become imbalanced due to various factors. When this is achieved and the tendency is toward promoting oxidation, oxidative stress is induced, causing potential cell damage and pathological diseases. Reactive oxygen species (ROS), which are the direct cause of this oxidative stress, are unstable and highly reactive, easily reacting with various biological substances, and attacking macromolecules in the body, causing irreversible damage to cells and tissues, mutations, and mutations. It causes cytotoxicity and carcinogenesis. Reactive nitrogen species (RNS), such as NO, HNO 2 , and ONOO-, are produced in large quantities due to the immune response of macrophages, neutrophils, and other immune cells during inflammatory reactions, and ROS is also produced at this time. The above-described free radicals oxidize and destroy cells in the body, thereby exposing them to various diseases.
본 발명의 다른 일실시예에서는, 산사나무 뿌리 추출물이 3T3-L1 지방 전구 세포에서 활성산소의 함량을 감소시킴을 확인하였다 (실시예 3 참조).In another example of the present invention, it was confirmed that hawthorn root extract reduces the content of active oxygen in 3T3-L1 preadipocytes (see Example 3).
본 발명의 또 다른 일실시예에서는, 산사나무 뿌리 추출물이 HFD 유도 비만 마우스의 지방 생성을 억제함을 확인하였다 (실시예 4 참조).In another example of the present invention, it was confirmed that hawthorn root extract inhibits fat production in HFD-induced obese mice (see Example 4).
본 발명의 용어 "비만(obesity)"은, 대사 장애로 인하여 체내에 지방세포가 증식 분화하고 이로 인하여 지방이 과잉으로 축적된 상태를 의미하며, 고혈압, 당뇨, 및 이상지질혈증 등을 동반하는 대사 증후군을 포함하는 관련 합병증을 유발할 수 있다. 에너지 흡수량이 소비량에 비해 상대적으로 증가하는 경우, 지방세포의 수와 부피가 증가되는 과정을 거쳐 결과적으로 지방조직의 질량이 증가된다.The term "obesity" of the present invention refers to a state in which fat cells proliferate and differentiate in the body due to metabolic disorders, resulting in excessive accumulation of fat, and is accompanied by high blood pressure, diabetes, and dyslipidemia. It can lead to associated complications, including syndromes. When energy absorption increases relative to consumption, the number and volume of fat cells increase, resulting in an increase in the mass of fat tissue.
본 발명의 용어, "백색 지방세포"는 중성 지방으로 대량의 지방 에너지를 체내에 축적하는 기능을 갖는 세포로서, 임신말기, 유아기, 사춘기에 집중하여 증식하며, 15배까지 부풀어 올라 지방 비대화 현상의 원인이 되기도 하며, 일반적으로 "지방세포"라 함은 "백색 지방세포"를 의미한다.As the term of the present invention, "white adipocytes" are cells with the function of accumulating large amounts of fat energy as neutral fat in the body, they proliferate intensively in late pregnancy, infancy, and puberty, and swell up to 15 times, causing the phenomenon of fat hypertrophy. It may also be the cause, and generally, “adipocytes” refer to “white fat cells.”
본 발명의 용어, "분화(differentiation)"는 세포가 분열 증식하여 성장하는 동안에 서로 구조나 기능이 특수화하는 현상, 즉 생물의 세포, 조직 등이 각각에게 주어진 일을 수행하기 위하여 형태나 기능이 변해가는 것을 말한다. 일반적으로 비교적 단순한 계(系)가 둘 이상의 질적으로 다른 부분계(部分系)로 분리되는 현상이다. 예를 들면, 개체 발생에서 처음에 동질적이었던 알 부분 사이에 머리나 몸통 등의 구별이 생기거나 세포에도 근세포 또는 신경세포 등의 구별이 생기는 것과 같이 처음에 거의 동질이었던 어떤 생물계의 부분 사이에 질적인 차이가 생기는 것, 또는 그 결과로서 질적으로 구별할 수 있는 부역 또는 부분계로 나누어져 있는 상태를 분화라고 한다.The term "differentiation" in the present invention refers to the phenomenon in which cells become specialized in structure or function while they divide and grow, that is, the cells, tissues, etc. of living organisms change their form or function to perform a given task. It says that In general, it is a phenomenon in which a relatively simple system is separated into two or more qualitatively different subsystems. For example, in ontogeny, distinctions such as the head or body arise between parts of an egg that were initially homogeneous, or distinctions such as muscle cells or nerve cells develop in cells, so qualitative changes occur between parts of a biological system that were initially almost homogeneous. The occurrence of personal differences, or the state of being divided into qualitatively distinguishable sub-regions or subsystems as a result, is called differentiation.
본 발명의 조성물 내의 상기 산사나무 뿌리 추출물의 함량은 질환의 증상, 증상의 진행 정도, 환자의 상태 등에 따라서 적절히 조절 가능하며, 예컨대, 전체 조성물 중량을 기준으로 0.0001 내지 99.9중량%, 또는 0.001 내지 50중량%일 수 있으나, 이에 한정되는 것은 아니다. 상기 함량비는 용매를 제거한 건조량을 기준으로 한 값이다.The content of the hawthorn root extract in the composition of the present invention can be appropriately adjusted depending on the symptoms of the disease, the degree of progression of the symptoms, the patient's condition, etc., for example, 0.0001 to 99.9% by weight, or 0.001 to 50%, based on the total weight of the composition. It may be weight percent, but is not limited thereto. The content ratio is a value based on the dry amount with the solvent removed.
본 발명에 따른 약학적 조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다. 상기 부형제는 예를 들어, 희석제, 결합제, 붕해제, 활택제, 흡착제, 보습제, 필름-코팅 물질, 및 제어방출첨가제로 이루어진 군으로부터 선택된 하나 이상일 수 있다.The pharmaceutical composition according to the present invention may further include appropriate carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions. The excipient may be, for example, one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, humectants, film-coating materials, and controlled-release additives.
본 발명에 따른 약학적 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 서방형 과립제, 장용과립제, 액제, 점안제, 엘실릭제, 유제, 현탁액제, 주정제, 트로키제, 방향수제, 리모나아데제, 정제, 서방형정제, 장용정제, 설하정, 경질캅셀제, 연질캅셀제, 서방캅셀제, 장용캅셀제, 환제, 틴크제, 연조엑스제, 건조엑스제, 유동엑스제, 주사제, 캡슐제, 관류액, 경고제, 로션제, 파스타제, 분무제, 흡입제, 패취제, 멸균주사용액, 또는에어로졸 등의 외용제 등의 형태로 제형화하여 사용될 수 있으며, 상기 외용제는 크림, 젤, 패치, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 또는 카타플라스마제 등의 제형을 가질 수 있다.The pharmaceutical composition according to the present invention can be prepared as powder, granules, sustained-release granules, enteric-coated granules, solutions, eye drops, ellipsis, emulsions, suspensions, spirits, troches, perfumes, and limonadese according to conventional methods. , tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric-coated capsules, pills, tinctures, soft extracts, dry extracts, liquid extracts, injections, capsules, perfusate, It can be formulated and used in the form of external preparations such as warning agents, lotions, pasta preparations, sprays, inhalants, patches, sterilized injection solutions, or aerosols, and the external preparations include creams, gels, patches, sprays, ointments, and warning agents. , it may have a dosage form such as lotion, liniment, pasta, or cataplasma.
본 발명에 따른 약학적 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 올리고당, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다.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, gum acacia, alginate, gelatin, and calcium. These include phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다.When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
본 발명에 따른 정제, 산제, 과립제, 캡슐제, 환제, 트로키제의 첨가제로 옥수수전분, 감자전분, 밀전분, 유당, 백당, 포도당, 과당, 디-만니톨, 침강탄산칼슘, 합성규산알루미늄, 인산일수소칼슘, 황산칼슘, 염화나트륨, 탄산수소나트륨, 정제 라놀린, 미결정셀룰로오스, 덱스트린, 알긴산나트륨, 메칠셀룰로오스, 카르복시메칠셀룰로오스나트륨, 카올린, 요소, 콜로이드성실리카겔, 히드록시프로필스타치, 히드록시프로필메칠셀룰로오스(HPMC), HPMC 1928, HPMC 2208, HPMC 2906, HPMC 2910, 프로필렌글리콜, 카제인, 젖산칼슘, 프리모젤 등 부형제; 젤라틴, 아라비아고무, 에탄올, 한천가루, 초산프탈산셀룰로오스, 카르복시메칠셀룰로오스, 카르복시메칠셀룰로오스칼슘, 포도당, 정제수, 카제인나트륨, 글리세린, 스테아린산, 카르복시메칠셀룰로오스나트륨, 메칠셀룰로오스나트륨, 메칠셀룰로오스, 미결정셀룰로오스, 덱스트린, 히드록시셀룰로오스, 히드록시프로필스타치, 히드록시메칠셀룰로오스, 정제쉘락, 전분호, 히드록시프로필셀룰로오스, 히드록시프로필메칠셀룰로오스, 폴리비닐알코올, 폴리비닐피롤리돈 등의 결합제가 사용될 수 있으며, 히드록시프로필메칠셀룰로오스, 옥수수전분, 한천가루, 메칠셀룰로오스, 벤토나이트, 히드록시프로필스타치, 카르복시메칠셀룰로오스나트륨, 알긴산나트륨, 카르복시메칠셀룰로오스칼슘, 구연산칼슘, 라우릴황산나트륨, 무수규산, 1-히드록시프로필셀룰로오스, 덱스트란, 이온교환수지, 초산폴리비닐, 포름알데히드처리 카제인 및 젤라틴, 알긴산, 아밀로오스, 구아르고무(Guar gum), 중조, 폴리비닐피롤리돈, 인산칼슘, 겔화전분, 아라비아고무, 아밀로펙틴, 펙틴, 폴리인산나트륨, 에칠셀룰로오스, 백당, 규산마그네슘알루미늄, 디-소르비톨액, 경질무수규산 등 붕해제; 스테아린산칼슘, 스테아린산마그네슘, 스테아린산, 수소화식물유(Hydrogenated vegetable oil), 탈크, 석송자, 카올린, 바셀린, 스테아린산나트륨, 카카오지, 살리실산나트륨, 살리실산마그네슘, 폴리에칠렌글리콜(PEG) 4000, PEG 6000, 유동파라핀, 수소첨가대두유(Lubri wax), 스테아린산알루미늄, 스테아린산아연, 라우릴황산나트륨, 산화마그네슘, 마크로골(Macrogol), 합성규산알루미늄, 무수규산, 고급지방산, 고급알코올, 실리콘유, 파라핀유, 폴리에칠렌글리콜지방산에테르, 전분, 염화나트륨, 초산나트륨, 올레인산나트륨, dl-로이신, 경질무수규산 등의 활택제;가 사용될 수 있다.Additives to tablets, powders, granules, capsules, pills, and troches according to the present invention include corn starch, potato starch, wheat starch, lactose, white sugar, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, and phosphoric acid. Calcium monohydrogen, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropylmethyl. Excipients such as cellulose (HPMC), HPMC 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethyl cellulose, sodium methyl cellulose, methyl cellulose, microcrystalline cellulose, dextrin. , hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, refined shellac, starch, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. binders can be used, Hydroxypropyl methyl cellulose, corn starch, agar powder, methyl cellulose, bentonite, hydroxypropyl starch, sodium carboxymethyl cellulose, sodium alginate, calcium carboxymethyl cellulose, calcium citrate, sodium lauryl sulfate, silicic anhydride, 1-hydroxy Propylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, Disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose, white sugar, magnesium aluminum silicate, di-sorbitol solution, light anhydrous silicic acid; Calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium, kaolin, petrolatum, sodium stearate, cacao fat, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogen. Added soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, Macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acids, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, Lubricants such as starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and light anhydrous silicic acid may be used.
본 발명에 따른 액제의 첨가제로는 물, 묽은 염산, 묽은 황산, 구연산나트륨, 모노스테아린산슈크로스류, 폴리옥시에칠렌소르비톨지방산에스텔류(트윈에스텔), 폴리옥시에칠렌모노알킬에텔류, 라놀린에텔류, 라놀린에스텔류, 초산, 염산, 암모니아수, 탄산암모늄, 수산화칼륨, 수산화나트륨, 프롤아민, 폴리비닐피롤리돈, 에칠셀룰로오스, 카르복시메칠셀룰로오스나트륨 등이 사용될 수 있다.Additives to the liquid according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.
본 발명에 따른 시럽제에는 백당의 용액, 다른 당류 혹은 감미제 등이 사용될 수 있으며, 필요에 따라 방향제, 착색제, 보존제, 안정제, 현탁화제, 유화제, 점조제 등이 사용될 수 있다.A solution of white sugar, other sugars, or sweeteners, etc. may be used in the syrup according to the present invention, and if necessary, flavoring agents, colorants, preservatives, stabilizers, suspending agents, emulsifiers, thickening agents, etc. may be used.
본 발명에 따른 유제에는 정제수가 사용될 수 있으며, 필요에 따라 유화제, 보존제, 안정제, 방향제 등이 사용될 수 있다.Purified water can be used in the emulsion according to the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. can be used as needed.
본 발명에 따른 현탁제에는 아카시아, 트라가칸타, 메칠셀룰로오스, 카르복시메칠셀룰로오스, 카르복시메칠셀룰로오스나트륨, 미결정셀룰로오스, 알긴산나트륨, 히드록시프로필메칠셀룰로오스(HPMC), HPMC 1828, HPMC 2906, HPMC 2910 등 현탁화제가 사용될 수 있으며, 필요에 따라 계면활성제, 보존제, 안정제, 착색제, 방향제가 사용될 수 있다.Suspensions according to the present invention include acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, etc. Topics may be used, and surfactants, preservatives, stabilizers, colorants, and fragrances may be used as needed.
본 발명에 따른 주사제에는 주사용 증류수, 0.9%염화나트륨주사액, 링겔주사액, 덱스트로스주사액, 덱스트로스+염화나트륨주사액, 피이지(PEG), 락테이티드 링겔주사액, 에탄올, 프로필렌글리콜, 비휘발성유-참기름, 면실유, 낙화생유, 콩기름, 옥수수기름, 올레인산에칠, 미리스트산 이소프로필, 안식향산벤젠과 같은 용제; 안식향산나트륨, 살리실산나트륨, 초산나트륨, 요소, 우레탄, 모노에칠아세트아마이드, 부타졸리딘, 프로필렌글리콜, 트윈류, 니정틴산아미드, 헥사민, 디메칠아세트아마이드와 같은 용해보조제; 약산 및 그 염(초산과 초산나트륨), 약염기 및 그 염(암모니아 및 초산암모니움), 유기화합물, 단백질, 알부민, 펩 톤, 검류와 같은 완충제; 염화나트륨과 같은 등장화제; 중아황산나트륨(NaHSO3)이산화탄소가스, 메타중아황산나트륨(Na2S2O5), 아황산나트륨(Na2SO3), 질소가스(N2), 에칠렌디아민테트라초산과 같은 안정제; 소디움비설파이드 0.1%, 소디움포름알데히드 설폭실레이트, 치오우레아, 에칠렌디아민테트라초산디나트륨, 아세톤소디움비설파이트와 같은 황산화제; 벤질알코올, 클로로부탄올, 염산프로카인, 포도당, 글루콘산칼슘과 같은 무통화제; 시엠시나트륨, 알긴산나트륨, 트윈 80, 모노스테아린산알루미늄과 같은 현탁화제를 포함할 수 있다.Injections according to the present invention include distilled water for injection, 0.9% sodium chloride injection, IV solution, dextrose injection, dextrose + sodium chloride injection, PEG, lactated IV solution, ethanol, propylene glycol, non-volatile oil - sesame oil. solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristic acid, and benzene benzoate; Solubilizing agents such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, Tween, nicotinic acid amide, hexamine, and dimethylacetamide; 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 buffering agents such as gums; Isotonic agents such as sodium chloride; Stabilizers such as sodium bisulfite (NaHSO3) carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; Sulfurizing agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, and acetone sodium bisulfite; Analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; It may contain suspending agents such as CM sodium, sodium alginate, Tween 80, and aluminum monostearate.
본 발명에 따른 좌제에는 카카오지, 라놀린, 위텝솔, 폴리에틸렌글리콜, 글리세로젤라틴, 메칠셀룰로오스, 카르복시메칠셀룰로오스, 스테아린산과 올레인산의 혼합물, 수바날(Subanal), 면실유, 낙화생유, 야자유, 카카오버터+콜레스테롤, 레시틴, 라네트왁스, 모노스테아린산글리세롤, 트윈 또는 스판, 임하우젠(Imhausen), 모놀렌(모노스테아린산프로필렌글리콜), 글리세린, 아뎁스솔리두스(Adeps solidus), 부티룸 태고-G(Buytyrum Tego-G), 세베스파마 16 (Cebes Pharma 16), 헥사라이드베이스 95, 코토마(Cotomar), 히드록코테 SP, S-70-XXA, S-70-XX75(S-70-XX95), 히드록코테(Hydrokote) 25, 히드록코테 711, 이드로포스탈 (Idropostal), 마사에스트라리움(Massa estrarium, A, AS, B, C, D, E, I, T), 마사-MF, 마수폴, 마수폴-15, 네오수포스탈-엔, 파라마운드-B, 수포시로(OSI, OSIX, A, B, C, D, H, L), 좌제기제 IV 타입 (AB, B, A, BC, BBG, E, BGF, C, D, 299), 수포스탈 (N, Es), 웨코비 (W, R, S, M ,Fs), 테제스터 트리글리세라이드 기제 (TG-95, MA, 57)와 같은 기제가 사용될 수 있다.Suppositories according to the present invention include cacao oil, lanolin, witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, lecithin, Lanet wax, glycerol monostearate, Tween or Span, Imhausen, monolene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydrocote SP, S-70-XXA, S-70-XX75(S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Massaupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), suppositories type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wecobi (W, R, S, M, Fs), Tegestor triglyceride base (TG-95, MA, 57) and The same mechanism can be used.
경구 투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스티레이트 탈크 같은 윤활제들도 사용된다.Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the extract with at least one excipient, such as starch, calcium carbonate, and sucrose. ) or prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate talc are also used.
경구 투여를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜 (propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다.Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.
본 발명에 따른 약학적 조성물은 약학적으로 유효한 양으로 투여한다. 본 발명에 있어서, "약학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 환자 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다.The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat the disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, activity of the drug, and the type of patient's disease. It can be determined based on factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other factors well known in the medical field.
본 발명에 따른 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 본 발명이 속하는 기술분야에 통상의 기술자에 의해 용이하게 결정될 수 있다.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 multiple times. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art to which the present invention pertains.
본 발명의 약학적 조성물은 개체에게 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구 복용, 피하 주사, 복강 투여, 정맥 주사, 근육 주사, 척수 주위 공간(경막내) 주사, 설하 투여, 볼점막 투여, 직장 내 삽입, 질 내 삽입, 안구 투여, 귀 투여, 비강 투여, 흡입, 입 또는 코를 통한 분무, 피부 투여, 경피 투여 등에 따라 투여될 수 있다.The pharmaceutical composition of the present invention can be administered to an individual through various routes. All modes of administration are contemplated, including oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, intrarectal injection, vaginal injection. It can be administered by internal insertion, ocular administration, otic administration, nasal administration, inhalation, spraying through the mouth or nose, dermal administration, transdermal administration, etc.
본 발명의 약학적 조성물은 치료할 질환, 투여 경로, 환자의 연령, 성별, 체중 및 질환의 중등도 등의 여러 관련 인자와 함께 활성성분인 약물의 종류에 따라 결정된다.The pharmaceutical composition of the present invention is determined depending on the type of drug as the active ingredient along with various related factors such as the disease to be treated, the route of administration, the patient's age, gender, weight, and severity of the disease.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물을 이를 필요로 하는 개체에 투여하는 단계를 포함하는, 지방간 또는 비만의 예방, 개선, 또는 치료 방법을 제공한다.Additionally, the present invention provides a method for preventing, improving, or treating fatty liver disease or obesity, comprising administering a hawthorn root extract or a composition containing the same as an active ingredient to an individual in need thereof.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 지방간 또는 비만의 예방, 개선, 또는 치료 용도를 제공한다.Additionally, the present invention provides the use of hawthorn root extract or a composition containing the same as an active ingredient to prevent, improve, or treat fatty liver disease or obesity.
또한, 본 발명은 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 지방간 또는 비만의 예방, 개선 또는 치료 제제를 제조하기 위한 용도를 제공한다.In addition, the present invention provides the use of hawthorn root extract or a composition containing it as an active ingredient for preparing a preparation for preventing, improving or treating fatty liver disease or obesity.
또한, 본 발명은 지방간 또는 비만의 예방, 개선, 또는 치료용 약제를 제조하기 위한 산사나무 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 용도를 제공한다.Additionally, the present invention provides the use of a hawthorn root extract or a composition containing the same as an active ingredient for preparing a medicine for preventing, improving, or treating fatty liver disease or obesity.
본 발명에서 "개체"란 질병의 치료를 필요로 하는 대상을 의미하고, 보다 구체적으로는 인간 또는 비-인간인 영장류, 생쥐 (mouse), 쥐 (rat), 개, 고양이, 말, 및 소 등의 포유류일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, “individual” refers to a subject in need of treatment for a disease, and more specifically, human or non-human primates, mice, rats, dogs, cats, horses, cows, etc. It may be a mammal, but is not limited thereto.
본 발명에서 "투여"란 임의의 적절한 방법으로 개체에게 소정의 본 발명의 조성물을 제공하는 것을 의미한다.In the present invention, “administration” means providing a given composition of the present invention to an individual by any suitable method.
본 발명에서 "예방"이란 목적하는 질환의 발병을 억제하거나 지연시키는 모든 행위를 의미하고, "치료"란 본 발명에 따른 약학적 조성물의 투여에 의해 목적하는 질환과 그에 따른 대사 이상 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미하며, "개선"이란 본 발명에 따른 조성물의 투여에 의해 목적하는 질환과 관련된 파라미터, 예를 들면 증상의 정도를 감소시키는 모든 행위를 의미한다.In the present invention, “prevention” refers to any action that suppresses or delays the onset of the desired disease, and “treatment” refers to the improvement or improvement of the desired disease and its associated metabolic abnormalities by administration of the pharmaceutical composition according to the present invention. It refers to all actions that are beneficially changed, and “improvement” refers to all actions that reduce parameters related to the target disease, such as the degree of symptoms, by administering the composition according to the present invention.
또한, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는, 지방간 또는 비만의 예방 또는 개선용 식품 조성물을 제공한다.Additionally, the present invention provides a food composition for preventing or improving fatty liver disease or obesity, comprising hawthorn root extract as an active ingredient.
본 발명에서, 산사나무, 산사나무 뿌리 추출물, 용매, 지방간, 조성물, 및 전사인자에 대하여 앞서 설명된 사항은 모두 본원의 지방간 또는 비만의 예방 또는 개선용 식품 조성물에 그대로 적용된다.In the present invention, all of the matters previously described regarding hawthorn, hawthorn root extract, solvent, fatty liver, composition, and transcription factor are directly applied to the food composition for preventing or improving fatty liver or obesity of the present application.
본 발명에 있어서, 식품 조성물은 건강기능성 식품 조성물일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the food composition may be a health functional food composition, but is not limited thereto.
또한, 산사나무 뿌리 추출물은 지방간 또는 비만의 예방 또는 개선을 목적으로 식품에 첨가될 수 있다.Additionally, hawthorn root extract can be added to foods for the purpose of preventing or improving fatty liver disease or obesity.
본 발명에 있어서 식품은 기능성 식품 및 건강기능성 식품을 포함한다. In the present invention, food includes functional foods and health functional foods.
본 발명의 산사나무 뿌리 추출물을 식품 첨가물로 사용할 경우, 상기 산사나무 뿌리 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용할 수 있다. 유효성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시 본 발명의 산사나무 뿌리 추출물은 원료에 대하여 15 중량% 이하, 또는 10 중량% 이하의 양으로 첨가될 수 있다. 그러나, 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.When using the hawthorn root extract of the present invention as a food additive, the hawthorn root extract can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). In general, when producing food or beverages, the hawthorn root extract of the present invention may be added in an amount of 15% by weight or less, or 10% by weight 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 may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다.There are no special restrictions on the types of foods above. Examples of foods to which the above substances can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, These include alcoholic beverages and vitamin complexes, and include all health functional foods in the conventional sense.
본 발명에 따른 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당 및 과당과 같은 모노사카라이드, 말토오스 및 수크로오스와 같은 디사카라이드, 덱스트린 및 시클로덱스트린과 같은 폴리사카라이드, 및 자일리톨, 소르비톨 및 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 mL당 일반적으로 약 0.01-0.20g, 또는 약 0.04-0.10g 이다.The health drink composition according to the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, like conventional drinks. The above-mentioned natural carbohydrates include 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, natural sweeteners such as thaumatin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used. The proportion of natural carbohydrates 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.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0.01-0.20 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, It may contain carbonating agents used in carbonated drinks. Additionally, the composition of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients can be used independently or in combination. The ratio of these additives is not very 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.
또한, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 화장료 조성물을 제공한다.Additionally, the present invention provides a cosmetic composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
본 발명에서, 산사나무, 산사나무 뿌리 추출물, 용매, 지방간, 조성물, 및 전사인자에 대하여 앞서 설명된 사항은 모두 본원의 비만의 예방 또는 개선용 화장료 조성물에 그대로 적용된다.In the present invention, all of the matters previously described regarding hawthorn, hawthorn root extract, solvent, fatty liver, composition, and transcription factor are directly applied to the cosmetic composition for preventing or improving obesity herein.
본 발명의 산사나무 뿌리 추출물은 중성지방의 축적을 억제하며, 셀룰라이트 감소 효과를 지니는 화장료 조성물에 첨가될 수 있다. 본 발명의 산사나무 뿌리 추출물을 첨가물로 사용할 경우, 산사나무 뿌리 추출물을 그대로 첨가하거나 다른 화장료 성분과 함께 사용할 수 있고, 통상의 방법에 따라 적절하게 사용될 수 있다. 유효성분의 혼합량은 사용목적에 따라 적합하게 결정될 수 있다.The hawthorn root extract of the present invention inhibits the accumulation of neutral fat and can be added to a cosmetic composition that has a cellulite reducing effect. When using the hawthorn root extract of the present invention as an additive, the hawthorn root extract can be added as is or used together with other cosmetic ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use.
본 발명의 일 구현예에 따른 화장료 조성물에 있어서, 상기 비만의 예방 또는 개선용 화장료 조성물은 스킨, 스킨 소프트너, 스킨 토너, 로션, 밀크로션, 모이스쳐로션, 영양로션, 마사지크림, 영양크림, 아이크림, 모이스쳐크림, 핸드크림, 에센스, 영양에센스, 팩, 클렌징폼, 클렌징워터, 클렌징크림, 바디로션, 바디클렌져, 비누 및 파우더 중에서 선택된 어느 하나의 제형을 가질 수 있으나, 이에 제한되지 않는다. 이들 각 제형으로 이루어진 화장료 조성물은 그 제형의 제제화에 필요하고 적절한 각종의 기제와 첨가물을 함유할 수 있다.In the cosmetic composition according to one embodiment of the present invention, the cosmetic composition for preventing or improving obesity is used for skin, skin softener, skin toner, lotion, milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, and eye cream. It may have a formulation selected from cream, moisture cream, hand cream, essence, nutritional essence, pack, cleansing foam, cleansing water, cleansing cream, body lotion, body cleanser, soap, and powder, but is not limited thereto. The cosmetic composition consisting of each of these formulations may contain various bases and additives necessary and appropriate for the formulation of the formulation.
본 발명의 화장료 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라가칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier ingredients. etc. can be used.
본 발명의 화장료 조성물의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판-부탄 또는 디메틸에테르와 같은 추진체를 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the cosmetic composition is a spray, chlorofluorohydride may be additionally used. It may contain propellants such as carbon, propane-butane or dimethyl ether.
본 발명의 화장료 조성물의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의지방산 에스테르가 있다.When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, solvating agent, or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, and propylene. These include glycols, 1,3-butylglycol oil, glycerol fatty esters, polyethylene glycol or fatty acid esters of sorbitan.
본 발명의 화장료 조성물의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a suspension, the carrier component includes water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
본 발명의 화장료 조성물의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르설페이트, 설포숙신산 모노에스테르, 아세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될수 있다.When the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyl taurate, and sarcosinate. , fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linoline derivative, or ethoxylated glycerol fatty acid ester can be used.
본 발명의 화장료 조성물은 형광물질, 살진균제, 굴수성 유발물질, 보습제, 방향제, 방향제 담체, 단백질, 용해화제, 당 유도체, 일광차단제, 비타민, 식물 추출물 등을 포함하는 부형제를 추가로 함유할 수 있다.The cosmetic composition of the present invention may further contain excipients including fluorescent substances, fungicides, hydrotropes-inducing substances, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, etc. .
또한, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 의약외품 조성물을 제공한다.In addition, the present invention provides a quasi-drug composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
본 발명에서, 산사나무, 산사나무 뿌리 추출물, 용매, 지방간, 조성물, 및 전사인자에 대하여 앞서 설명된 사항은 모두 본원의 비만의 예방 또는 개선용 의약외품 조성물에 그대로 적용된다.In the present invention, all of the matters previously described regarding hawthorn, hawthorn root extract, solvent, fatty liver, composition, and transcription factor are directly applied to the quasi-drug composition for preventing or improving obesity herein.
본 발명에서 용어 "의약외품"은 사람이나 동물의 질병을 치료, 경감, 처치 또는 예방할 목적으로 사용되는 섬유, 고무제품 또는 이와 유사한 것, 인체에 대한 작용이 약하거나 인체에 직접 작용하지 아니하며, 기구 또는 기계가 아닌 것과 이와 유사한 것, 감염형 예방을 위하여 살균, 살충 및 이와 유사한 용도로 사용되는 제제 중 하나에 해당하는 물품으로서, 사람이나 동물의 질병을 진단, 치료, 경감, 처치 또는 예방할 목적으로 사용하는 물품 중 기구, 기계 또는 장치가 아닌 것 및 사람이나 동물의 구조와 기능에 약리학적 영향을 줄 목적으로 사용 하는 물품 중 기구, 기계 또는 장치가 아닌 것을 제외한 물품을 의미한다.In the present invention, the term "quasi-drug" refers to fibers, rubber products, or similar products used for the purpose of treating, alleviating, treating, or preventing diseases in humans or animals. They have a weak effect on the human body or do not directly act on the human body, and refer to devices or Items that are non-machine or similar, or products that are used for sterilization, insecticide and similar purposes to prevent infectious diseases, and are used for the purpose of diagnosing, treating, mitigating, treating or preventing diseases in humans or animals. It refers to articles that are not instruments, machines, or devices among articles used for the purpose of having a pharmacological effect on the structure and function of humans or animals, excluding articles that are not instruments, machines, or devices.
본 발명의 조성물을 의약외품 조성물로 사용할 경우, 상기 산사나무 뿌리 추출물을 그대로 첨가하거나 다른 의약외품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 본 발명의 일 구현예에 따르면 본 발명의 의약외품 조성물은 소독청결제, 치약, 샤워폼, 가그린, 물티슈, 세제, 비누, 핸드워시, 가습기 충진제, 마스크, 연고제 또는 필터충진제일 수 있다.When using the composition of the present invention as a quasi-drug composition, the hawthorn root extract can be added as is or used together with other quasi-drug ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). According to one embodiment of the present invention, the quasi-drug composition of the present invention may be a disinfectant cleaner, toothpaste, shower foam, gargle, wet tissue, detergent, soap, hand wash, humidifier filler, mask, ointment, or filter filler.
또한, 본 발명은 산사나무 뿌리 추출물을 유효성분으로 포함하는, 시험관 내(in vitro) 지방세포 분화 억제용 조성물을 제공한다.Additionally, the present invention provides a composition for inhibiting adipocyte differentiation in vitro, comprising hawthorn root extract as an active ingredient.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시한다. 그러나 하기의 실시예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 하기 실시예에 의해 본 발명의 내용이 한정되는 것은 아니다.Below, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are provided only to make the present invention easier to understand, and the content of the present invention is not limited by the following examples.
실험예 1. 산사나무 뿌리 추출물의 준비Experimental Example 1. Preparation of hawthorn root extract
산사나무(Crataegus pinnatifida) 뿌리 추출물은 한국 식물 추출물 은행 (충청북도 오창읍)에서 분양받았다. 추출물은 100% 메탄올로 추출되었으며, 추출 후 추출액을 여과하고 진공 증발 건조 시켰다. 시험관 실험에 사용하기 위하여 획득한 추출물을 디메틸 설폭사이드(dimethyl sulfoxide, 이하 'DMSO')에 50mg/ml로 용해시킨 후 희석하여 사용하였다. 전임상실험의 경우 획득한 추출물을 멸균수에 용해시켜 사용하였다.Hawthorn ( Crataegus pinnatifida ) root extract was obtained from the Korea Plant Extract Bank (Ochang-eup, Chungcheongbuk-do). The extract was extracted with 100% methanol, and after extraction, the extract was filtered and vacuum evaporated to dryness. To be used in test tube experiments, the obtained extract was dissolved in dimethyl sulfoxide (DMSO) at 50 mg/ml and then diluted. For preclinical experiments, the obtained extract was dissolved in sterile water and used.
실험예 2. 동물 및 산사나무 뿌리 추출물 (CRE) 투여Experimental Example 2. Animals and Hawthorn Root Extract (CRE) Administration
6 주령 수컷 C57BL/6 마우스는 효창 사이언스 (대한민국, 대구)에서 구입하였다. 마우스는 순응을 위해 25-30℃에서 12 시간의 명암 주기에서 2 주 동안 우리에서 사육하였다. 2 주일의 순응 후, 마우스를 식이에 따라 무작위로 5 개의 그룹 (n=7)으로 나누었다; (1) 정상적인 식단 (ND); (2) 고지방 식단(HFD); (3) HFD에 CRE를 1.25 mg/kg/일로 투여; (4) HFD에 CRE를 2.5 mg/kg/일로 투여 및 (5) HFD에 CRE를 5 mg/kg/일로 투여. 음식과 물은 12주동안 무제한으로 제공하였다. 체중과 식이섭취는 2일에 한 번 측정되었고, 추출물은 경구투여로 2일에 한 번 투여하였다. 동물 실험 마지막 날 장기와 혈액을 채취하여 분석에 이용하였다. 동물 실험은 대한민국 대구의 경북대학교 동물병원 동물보호위원회의 승인을 받았다 (승인번호 : KNU 2022-0125). Six-week-old male C57BL/6 mice were purchased from Hyochang Science (Daegu, Korea). Mice were housed in cages for 2 weeks under a 12-hour light/dark cycle at 25-30°C for acclimatization. After 2 weeks of acclimatization, mice were randomly divided into five groups (n=7) according to diet; (1) normal diet (ND); (2) high-fat diet (HFD); (3) CRE administered on HFD at 1.25 mg/kg/day; (4) CRE administered on a HFD at 2.5 mg/kg/day and (5) CRE administered on a HFD at 5 mg/kg/day. Food and water were provided ad libitum for 12 weeks. Body weight and dietary intake were measured once every two days, and the extract was administered orally once every two days. On the last day of the animal experiment, organs and blood were collected and used for analysis. Animal experiments were approved by the Animal Care Committee of Kyungpook National University Animal Hospital, Daegu, Korea (approval number: KNU 2022-0125).
실험예 3. 조직 헤마톡실린 및 에오신(H&E) 염색Experimental Example 3. Tissue hematoxylin and eosin (H&E) staining
마우스로부터 서혜부 백색 지방 조직 (inguinal white adipose tissue 이하 'iWAT'), 부고환 백색 지방 조직 (epididymal white adipose tissue 이하 'eWAT') 및 간(liver)을 적출한 뒤, 4% 파라포름알데히드(parpaformaldehyde)에 1일 동안 보관하여 조직을 고정한 뒤, 조직을 파라핀에 포매하고 5μm 두께로 절단한 뒤 표준 프로토콜에 따라 H & E로 염색한 다음 현미경으로 검사하였다.Inguinal white adipose tissue ('iWAT'), epididymal white adipose tissue ('eWAT'), and liver were extracted from mice and then placed in 4% paraformaldehyde. After storing for 1 day and fixing the tissue, the tissue was embedded in paraffin, cut into 5 μm thick sections, stained with H & E according to standard protocols, and then examined under a microscope.
실험예 4. 혈액 생화학 분석Experimental Example 4. Blood biochemical analysis
동물실험 12주 마지막 날 희생하였으며, 휙득한 혈액은 3,500rpm에서 15분간 원심분리하여 혈장을 분리해내었다. 혈장 내 AST (Aspartate aminotrasnferase) 및 ALT (Alanine aminotransferase) 수준은 Olympus AU 400 (Olympus Optical, Tokyo, Japan) 분석기를 사용하여 제조업체의 절차에 따라 측정하였다.Animals were sacrificed on the last day of the 12th week of the experiment, and the collected blood was centrifuged at 3,500 rpm for 15 minutes to separate plasma. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in plasma were measured using an Olympus AU 400 (Olympus Optical, Tokyo, Japan) analyzer according to the manufacturer's procedures.
실험예 5. 항산화 분석Experimental Example 5. Antioxidant analysis
3T3-L1 지방 전구 세포를 6-웰 플레이트에 시딩하였으며 8일 분화 후 DCFDA/H2DCFDA-Celluar ROS Assay Kit (Abcam, US)를 이용해 형광 현미경 및 유세포 분석기를 사용하여 공급업체의 프로토콜을 참고하여 분석하였다.3T3-L1 preadipocytes were seeded in 6-well plates, and after 8 days of differentiation, they were analyzed using a fluorescence microscope and flow cytometer using the DCFDA/H2DCFDA-Celluar ROS Assay Kit (Abcam, US), referring to the supplier's protocol. .
DPPH (2,2-diphenyl-1-picrylhydrazyl) 라디칼 소거활성 능력 측정은 CRE를 DMSO로 2, 4, 6, 8, 10μg/ml로 희석한 후 볼텍싱 하고, DPPH 10.0039g을 에탄올에 희석하여 DPPH 시약을 준비하였다. 이후 아스코르브산(Ascorbic acid)를 표준물질로 이용하고 시약에 볼텍싱한 CRE 또는 아스코르브산을 상기 농도에 맞게 주입한 후 30분 간 암반응 시켰다. 그 후, 517nm에서 흡광도를 3회 반복으로 측정하였다. To measure DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, CRE was diluted with DMSO to 2, 4, 6, 8, and 10 μg/ml, then vortexed, and 10.0039 g of DPPH was diluted in ethanol to obtain DPPH. Reagents were prepared. Afterwards, ascorbic acid was used as a standard material, and CRE or ascorbic acid vortexed into the reagent was injected at the above concentration and reacted in the dark for 30 minutes. Afterwards, the absorbance was measured at 517 nm three times.
ABTS radical decolorization (2,2'-azinobis-3-ehtlbezothiazoline-6-sulfonic acid) 측정은 CRE를 DMSO로 2, 4, 6, 8, 10μg/ml로 희석한 후 볼텍싱 하고, ABTS에 과황산칼륨(potassium persulfate)를 첨가하여 증류수에 용해시켜 실온에 12 시간 방치하여 ABTS 시약을 준비하였다. 이 후 아스코르브산을 표준 물질로 이용하여 볼텍싱한 CRE 또는 아스코르브산을 상기 농도에 맞게 ABTS 시약에 주입한 후 30분간 암반응 시켰다. 그 후 734nm에서 흡광도를 3회 반복으로 측정하였다.To measure ABTS radical decolorization (2,2'-azinobis-3-ehtlbezothiazoline-6-sulfonic acid), CRE was diluted with DMSO to 2, 4, 6, 8, and 10 μg/ml, then vortexed, and ABTS was added with potassium persulfate. (potassium persulfate) was added, dissolved in distilled water, and left at room temperature for 12 hours to prepare the ABTS reagent. Afterwards, CRE or ascorbic acid vortexed using ascorbic acid as a standard substance was injected into the ABTS reagent according to the above concentration and reacted in the dark for 30 minutes. Afterwards, the absorbance was measured three times at 734 nm.
실험예 6. 혈액 생화학 분석Experimental Example 6. Blood biochemical analysis
동물실험 12주 마지막 날 희생하였으며, 휙득한 혈액은 3,500rpm에서 15분간 원심분리하여 혈장을 분리해내었다. 혈장 내 AST (Aspartate aminotrasnferase), ALT (Alanine aminotransferase), ALP (Alkaline phosphatase), TP (Total protein), ALB (Albumin), T-BIL (total bilirubin), D-BIL(Direct bilirubin), BUN (Blood Urea Nitrogen), CREA (creatinine), TG (트리글리세라이드) 및 CHO (콜레스테롤) 수준은 Olympus AU 400 (Olympus Optical, Tokyo, Japan) 분석기를 사용하여 제조업체의 절차에 따라 측정하였다.Animals were sacrificed on the last day of the 12th week of the experiment, and the collected blood was centrifuged at 3,500 rpm for 15 minutes to separate plasma. AST (Aspartate aminotransferase), ALT (Alanine aminotransferase), ALP (Alkaline phosphatase), TP (Total protein), ALB (Albumin), T-BIL (total bilirubin), D-BIL (Direct bilirubin), BUN (Blood) in plasma Urea nitrogen (CREA), creatinine (CREA), triglyceride (TG), and cholesterol (CHO) levels were measured using an Olympus AU 400 (Olympus Optical, Tokyo, Japan) analyzer according to the manufacturer's procedures.
실험예 7. 지방 세포 분화Experimental Example 7. Adipocyte differentiation
3T3-L1 지방 전구 세포는 10% Bovine calf serum(BCS)가 보충된 둘베코의 변형된 이글 배지(DMEM)에서 유지되었다. 3T3-L1 지방 전구 세포를 100파이 실험접시에 컨플루언스가 80%가 되면 6-웰 플레이트에 시딩하고 2 일 동안 컨플루언스에 도달하도록 성장시켰다. 컨플루언스 후 2 일째에, 지방 세포 분화를 시작하기 위해, 배지를 0.5 mM 3-이소부틸-1-메틸크산틴 (IBMX), 0.25 mM 덱사메타손 (DEX), 1 μg/mL 인슐린, 0.125 mM 인도메타신을 함유하는 분화 배지-I(Differentiaion medium I, DMI)로 변경하였다. 그 후, 분화 배지-II(Differentiaion medium II, DMII)로 6일간 지방세포의 성숙을 유도하여 총 8일간 분화 하였다. 3T3-L1 preadipocytes were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% bovine calf serum (BCS). 3T3-L1 preadipocytes were seeded in a 6-well plate when confluence reached 80% in a 100-pi experimental dish and grown to reach confluence for 2 days. At 2 days after confluence, to initiate adipocyte differentiation, medium was supplemented with 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.25 mM dexamethasone (DEX), 1 μg/mL insulin, 0.125 mM India. It was changed to Differentiation medium I (DMI) containing metacin. Afterwards, maturation of adipocytes was induced for 6 days with differentiation medium II (DMII), and differentiation was performed for a total of 8 days.
실험예 8. 오일 레드 O 염색Experimental Example 8. Oil Red O staining
3T3-L1 지방 전구 세포를 CRE의 존재 또는 부재 하에 분화하도록 유도하였다. 분화 8일 차의 지방 세포를 인산 완충 식염수(PBS)로 세척하고, 파라포름알데히드로 고정시킨 뒤 여과된 0.3% 오일 레드 O 용액으로 30분간 상온에서 염색하였다. 이어서, 반응 생성물을 이소프로필알코올에 용해시켰다. 반응의 흡광도는 450 nm에서 측정되었다. 3T3-L1 preadipocytes were induced to differentiate in the presence or absence of CRE. Adipocytes on day 8 of differentiation were washed with phosphate-buffered saline (PBS), fixed with paraformaldehyde, and stained with filtered 0.3% Oil Red O solution for 30 minutes at room temperature. The reaction product was then dissolved in isopropyl alcohol. The absorbance of the reaction was measured at 450 nm.
실험예 9. 세포 생존율 분석 Experimental Example 9. Cell viability analysis
3T3-L1 지방 전구 세포를 96-웰 플레이트에 시딩 하였다. 배양한 후, 세포를 48시간 동안 CRE 또는 DMSO로 처리하였다. MTT(3-(4,5)-디메틸티아조-2-일-2,5-디페닐테트라졸륨 브로마이드) 용액을 각 웰에 첨가하고 4시간 동안 37℃에서 인큐베이션 하였다. 반응 생성물을 이소프로필알코올에 용해시켰다. 반응의 흡광도는 595 nm에서 측정되었다.3T3-L1 preadipocytes were seeded in 96-well plates. After culturing, cells were treated with CRE or DMSO for 48 hours. MTT (3-(4,5)-dimethylthiazo-2-yl-2,5-diphenyltetrazolium bromide) solution was added to each well and incubated at 37°C for 4 hours. The reaction product was dissolved in isopropyl alcohol. The absorbance of the reaction was measured at 595 nm.
실험예 10. 트리글리세라이드(TG) 분석Experimental Example 10. Triglyceride (TG) analysis
3T3-L1 지방 전구 세포를 CRE의 존재 또는 부재 하에 분화하도록 유도하였다. 분화 8일 차의 지방 세포를 인산 완충 식염수(PBS)로 세척하였고, 세포를 5% NP-40 용해 완충액에 용해시켰다. 이 후 Triglyceride assay kit를 사용하여 제조업체의 절차에 따라 측정하였다.3T3-L1 preadipocytes were induced to differentiate in the presence or absence of CRE. Adipocytes on day 8 of differentiation were washed with phosphate-buffered saline (PBS), and the cells were dissolved in 5% NP-40 lysis buffer. Afterwards, it was measured using a Triglyceride assay kit according to the manufacturer's procedure.
실험예 11. 웨스턴 블롯 분석 Experimental Example 11. Western blot analysis
분화 8일차의 3T3-L1 지방 전구 세포를 포스파타제 억제제 및 프로테아제 억제제를 함유하는 방사선 면역 침전 분석(RIPA) 용해 완충액으로 용해시켰다. 총 단백질(30 ㎍) 함량을 전기 영동을 통해 10% SDS-폴리아크릴아미드 겔에서 분리하고 니트로셀룰로스 막으로 옮겼다. 막을 1시간 동안 PBS 중의 5% 탈지유로 차단하고, 관련 1차 항체와 함께 4℃에서 밤새 인큐베이션한 후, 막을 TBS-T 완충액으로 10분동안 3회 세척하였다. 이어서 막을 HRP-접합된 이차 항체와 함께 1시간 동안 반응시켰다. 단백질 밴드를 ECL을 사용하여 가시화하고 퓨전 솔로 검출기(Vilber Lourmat, Marne La Vallee, France)에서 검출하였고, β-액틴에 대하여 표준화함으로써 평가되었다.3T3-L1 preadipocytes on day 8 of differentiation were lysed with radioimmunoprecipitation assay (RIPA) lysis buffer containing phosphatase inhibitors and protease inhibitors. Total protein (30 μg) content was separated on a 10% SDS-polyacrylamide gel by electrophoresis and transferred to a nitrocellulose membrane. Membranes were blocked with 5% skim milk in PBS for 1 hour, incubated with relevant primary antibodies overnight at 4°C, and then membranes were washed three times for 10 minutes with TBS-T buffer. The membrane was then reacted with HRP-conjugated secondary antibody for 1 hour. Protein bands were visualized using ECL, detected on a Fusion Solo detector (Vilber Lourmat, Marne La Vallee, France), and evaluated by normalizing to β-actin.
실험예 12. 실시간 역전사 폴리머라제 연쇄 반응(RT-PCR)Experimental Example 12. Real-time reverse transcription polymerase chain reaction (RT-PCR)
3T3-L1 지방 전구 세포를 6-웰 플레이트에 시딩하여 CRE의 존재 여부에 따라 그룹을 나누어 분화하였다. 분화 8일차의 3T3-L1 세포를 PBS로 세척한 후, 트리졸(Trizol) 시약을 사용하여 세포로부터 총 RNA를 추출하였다. cDNA 라이브러리는 PrimeScriptTM RT 시약 키트(TaKaRa Bio)로 합성하였다. mRNA 발현 수준은 SYBR Green(TOYOBO, Japan)을 사용하여 iCycler iQTM Real-Time PCR Detection System(Bio-Rad Laboratories, USA)에 의해 구현된 cDNA의 분석에 의해 정량화되었고, β-액틴에 대하여 표준화하였으며 상대적 수준의 유전자 발현을 배수로 표시하였다.3T3-L1 preadipocytes were seeded in a 6-well plate and differentiated into groups according to the presence or absence of CRE. 3T3-L1 cells on day 8 of differentiation were washed with PBS, and then total RNA was extracted from the cells using Trizol reagent. The cDNA library was synthesized using the PrimeScript TM RT reagent kit (TaKaRa Bio). mRNA expression levels were quantified by analysis of cDNA implemented by the iCycler iQTM Real-Time PCR Detection System (Bio-Rad Laboratories, USA) using SYBR Green (TOYOBO, Japan), normalized to β-actin, and relative Levels of gene expression were expressed as folds.
실험예 13. 통계 분석Experimental Example 13. Statistical Analysis
모든 실험은 3회 반복으로 진행하였으며, 통계 분석 데이터는 평균±SEM 및 SD로 표시되고 일원 분산 분석을 사용하여 분석되었다. 0.05 미만의 p-값은 통계적으로 유의한 것으로 간주되었다.All experiments were performed in triplicate, and statistical analysis data were expressed as mean ± SEM and SD and analyzed using one-way analysis of variance. A p-value of less than 0.05 was considered statistically significant.
실시예 1. 산사나무 뿌리 추출물(CRE)의 3T3-L1 지방 전구세포에서 지방 세포 분화 억제 효과 확인Example 1. Confirmation of the inhibitory effect of hawthorn root extract (CRE) on adipocyte differentiation in 3T3-L1 preadipocytes.
3T3-L1 지방 전구 세포에 CRE를 처리하였을 때 지방 세포 분화 억제 효과를 조사하였다. 먼저, 상기 실험예 9의 방법으로 세포 생존율을 분석한 결과, 도 1a에 나타난 바와 같이, CRE는 3T3-L1 세포의 생존율에 영향을 끼치지 않는 점 또는 3T3-L1 세포에 세포 독성을 야기하지 않는 점을 확인하였다. The inhibitory effect on adipocyte differentiation was investigated when 3T3-L1 preadipocytes were treated with CRE. First, as a result of analyzing the cell viability by the method of Experiment 9, as shown in Figure 1a, CRE does not affect the survival rate of 3T3-L1 cells or does not cause cytotoxicity in 3T3-L1 cells. was confirmed.
또한, 상기 실험예 8의 방법으로 오일 레드 O 염색을 통해 지방 함량을 분석한 결과, 도 1b에 나타난 바와 같이, 3T3-L1 세포 성숙 과정에서 CRE를 투여한 그룹은 지방 함량이 감소하는 점을 확인하였고, 이는 CRE 농도 의존적으로 감소하는 점을 확인하였다.In addition, as a result of analyzing the fat content through Oil Red O staining using the method of Experimental Example 8, it was confirmed that the fat content of the group administered CRE during the 3T3-L1 cell maturation process decreased, as shown in Figure 1b. It was confirmed that this decreases in a CRE concentration-dependent manner.
또한, 상기 실험예 10을 통해 상기 현상이 3T3-L1 세포의 트리글리세라이드(TG) 함량에도 영향을 미치는지 확인해본 결과, 도 1c에 나타난 바와 같이, CRE 처리시 농도 의존적으로 세포내 TG 함량이 감소하는 점을 확인하였다.In addition, as a result of confirming whether the above phenomenon also affects the triglyceride (TG) content of 3T3-L1 cells through Experimental Example 10, as shown in Figure 1c, the intracellular TG content decreases in a concentration-dependent manner upon CRE treatment. The point was confirmed.
또한, 상기 현상이 분자 수준에서도 재현이 되는지 확인해 보기 위해 상기 실험예 11 및 12의 방법으로 지방 세포 분화 관련 유전자의 발현을 확인해보았다.Additionally, in order to confirm whether the above phenomenon could be reproduced at the molecular level, the expression of genes related to adipocyte differentiation was checked using the methods of Experimental Examples 11 and 12 above.
도 1d에 나타난 바와 같이, CRE를 처리하였을 때 지방 세포 분화에 관여하는 유전자인 아디포넥틴, C/EBPα, PPARγ. SREBP-1의 mRNA발현이 농도 의존적으로 감소하는 점을 확인하였고, 도 1e에 나타난 바와 같이, CRE를 처리하였을 때 C/EBPα, PPARγ의 단백질 발현도 감소한 점을 확인하였다.As shown in Figure 1d, adiponectin, C/EBPα, and PPARγ, which are genes involved in adipocyte differentiation when treated with CRE. It was confirmed that the mRNA expression of SREBP-1 decreased in a concentration-dependent manner, and as shown in Figure 1e, the protein expression of C/EBPα and PPARγ also decreased when CRE was treated.
종합하면, 이러한 결과는 CRE는 세포 독성을 야기하지 않고 지방 전구 세포 분화 억제를 통한 항비만 효과를 나타내는바, 지방간 또는 비만의 개선 또는 치료에 이용될 수 있음을 시사한다.Taken together, these results suggest that CRE does not cause cytotoxicity and exhibits an anti-obesity effect through inhibition of adipocyte differentiation, suggesting that it can be used to improve or treat fatty liver or obesity.
실시예 2. HFD 유도된 비만 마우스에서 CRE의 지방증 저해 효과 및 간 손상 개선 효과Example 2. Effect of CRE on steatosis inhibition and liver damage improvement in HFD-induced obese mice
CRE의 생체 내에서의 지방증 저해 효과를 확인하고자, HFD 유도된 비만 마우스의 iWAT, eWAT, 간에서 CRE 투여를 통한 지방구 크기 변화를 조사하였다.To confirm the in vivo steatosis inhibition effect of CRE, changes in fat globule size were investigated in iWAT, eWAT, and liver of HFD-induced obese mice by CRE administration.
상기 실험예 3의 방법으로 마우스의 iWAT, eWAT 및 간 조직을 H&E로 염색하고 관찰한 결과, 도 2a에 나타난 바와 같이, HFD 대비 CRE 처리군에서 지방구의 크기가 유의하게 감소한 것을 확인하였다.As a result of staining and observing iWAT, eWAT, and liver tissue of mice with H&E using the method of Experimental Example 3, it was confirmed that the size of fat globules was significantly reduced in the CRE-treated group compared to HFD, as shown in Figure 2a.
또한, 상기 결과가 CRE의 지방세포 분화 관련 유전자 조절에 기인한 것인지 확인해보았다.In addition, we confirmed whether the above results were due to CRE's regulation of genes related to adipocyte differentiation.
그 결과 도 2b에 나타난 바와 같이, iWAT에서 CRE 처리를 통해 SREBP-1의 발현이 감소한 것을 확인하였고, SREBP-1의 하류영역인 PPARγ의 발현 또한 감소한 것을 확인하였다.As a result, as shown in Figure 2b, it was confirmed that the expression of SREBP-1 was decreased in iWAT through CRE treatment, and the expression of PPARγ, a downstream region of SREBP-1, was also confirmed to be decreased.
또한, 앞서 CRE 처리를 통해 간에서 지방구의 크기가 감소한 것을 확인한 바, CRE의 HFD로 유도된 간 손상의 개선 효과 여부를 확인하였다.In addition, as it was previously confirmed that the size of fat globules in the liver was reduced through CRE treatment, the effectiveness of CRE in improving liver damage induced by HFD was confirmed.
그 결과 도 2c에 나타난 바와 같이, CRE를 처리할 경우 HFD로 유도된 간의 손상된 형태가 개선되는 것을 확인하였다.As a result, as shown in Figure 2c, it was confirmed that treatment with CRE improved the damaged form of the liver induced by HFD.
또한, 상기 실험예 4의 방법으로 상기 간 손상의 개선을 AST, ALT 지표를 통해서도 확인해본 결과, 도 2d에 나타난 바와 같이, 혈장 내 AST, ALT 수치도 CRE 처리시 유의적으로 감소한 것을 확인하였으며 특히 ALT의 경우 그 감소폭이 큰 것을 확인하였다. In addition, as a result of confirming the improvement of liver damage through AST and ALT indicators using the method of Experimental Example 4, it was confirmed that AST and ALT levels in plasma were significantly reduced upon CRE treatment, as shown in Figure 2d. In the case of ALT, it was confirmed that the decrease was large.
상기 결과는 CRE가 HFD로 유도된 비만 마우스에서 간의 지방증 저해 및 HFD로 유도된 간 손상을 개선하는데 도움이 됨을 시사한다.The above results suggest that CRE is helpful in inhibiting liver steatosis and improving HFD-induced liver damage in HFD-induced obese mice.
실시예 3. CRE의 항산화 효과Example 3. Antioxidant effect of CRE
지방 분해시, 세포내 TG를 분해하면 지방산 산화로 이어지며 지방산 산화가 알어나면 세포내 지방 함량이 감소하면서 생체내에 에너지를 공급하게 된다.During fat decomposition, breaking down intracellular TG leads to fatty acid oxidation, and when fatty acid oxidation occurs, intracellular fat content decreases and energy is supplied to the body.
하지만 지방산 산화를 유도할 경우 단백질 손상, 지질 과산화, DNA 손상이 야기되며 세포내 산화 스트레스를 유도하여 생체 내 염증을 야기하는 바, 항비만 효과를 가지는 조성물의 항산화 효과는 매우 중요하다.However, inducing fatty acid oxidation causes protein damage, lipid peroxidation, and DNA damage, and induces intracellular oxidative stress, causing inflammation in vivo, so the antioxidant effect of a composition with an anti-obesity effect is very important.
이에, CRE가 항산화 효과를 갖는지 확인해보고자 상기 실험예 5의 방법으로 3T3-L1 지방 전구 세포에 CRE 처리시 세포내 활성산소(ROS) 함량의 변화를 조사해보았다.Therefore, to confirm whether CRE has an antioxidant effect, changes in intracellular reactive oxygen species (ROS) content were examined when 3T3-L1 preadipocytes were treated with CRE using the method of Experimental Example 5 above.
도 3a에 나타난 바와 같이, 3T3-L1 세포의 분화 과정중 CRE를 처리하였을 때 세포내 ROS 함량이 유의적으로 감소하는 것을 형광 현미경을 통해 확인하였다.As shown in Figure 3a, it was confirmed through fluorescence microscopy that the intracellular ROS content was significantly reduced when CRE was treated during the differentiation of 3T3-L1 cells.
또한, 도 3b에 나타난 바와 같이, 유세포 분석을 통해 세포내 ROS의 감소를 정량적으로 확인하였고, 이를 통해 CRE 처리시 유도된 산화 스트레스가 감소한 것을 확인하였다. In addition, as shown in Figure 3b, the decrease in intracellular ROS was quantitatively confirmed through flow cytometry, and through this, it was confirmed that oxidative stress induced during CRE treatment was reduced.
또한, 표 1에 나타난 바와 같이 CRE의 라디칼 소거능을 IC50을 통해 평가해본 결과 ABTS assay에서는 에서는 23.674±1.452μg/mL, DPPH assay에서는 24.618±0.254μg/mL로 나타났으며, 이는 대조군인 Ascorbic acid와 비교했을 때, 약 4배 높은 수치이며 라디칼 소거능이 Ascorbic acid의 1/4로 매우 높은 수준임을 확인하였다.In addition, as shown in Table 1, the radical scavenging ability of CRE was evaluated through IC50 and was found to be 23.674 ± 1.452 μg/mL in the ABTS assay and 24.618 ± 0.254 μg/mL in the DPPH assay, which is similar to that of Ascorbic acid, which is the control group. When compared, it was confirmed that the value was about 4 times higher and that the radical scavenging ability was very high at 1/4 of that of ascorbic acid.
Figure PCTKR2023019426-appb-img-000001
Figure PCTKR2023019426-appb-img-000001
종합하면, CRE는 지방 분해시 발생하는 산화 스트레스를 해소하여 산화 스트레스로 인한 세포 손상을 방지할 수 있음을 시사한다.Taken together, this suggests that CRE can prevent cell damage caused by oxidative stress by relieving oxidative stress that occurs during lipolysis.
실시예 4. HFD 유도된 비만 마우스에서 CRE의 체중 감소 효과Example 4. Weight loss effect of CRE in HFD-induced obese mice
In vitro 실험을 통해 CRE의 지방 세포 분화 억제를 통한 지방 생성 억제 효과를 확인한 바, 생체 내에서의 지방 생성 억제 효과를 확인하고자, 상기 실험예 2의 방법으로 HFD 유도된 비만 마우스로 in vivo에서의 실험을 진행하였다.Through in vitro experiments, the inhibitory effect of CRE on adipogenesis through inhibition of adipocyte differentiation was confirmed. To confirm the inhibitory effect on adipogenesis in vivo, HFD-induced obese mice were used in vivo using the method of Experimental Example 2. An experiment was conducted.
먼저, 도 4a에 나타난 바와 같이, CRE 투여는 HFD 유도된 비만 마우스의 사료 섭취량에 영향을 미치지 않은 점을 확인하였다.First, as shown in Figure 4a, it was confirmed that CRE administration did not affect the food intake of HFD-induced obese mice.
또한, 도 4b에 나타난 바와 같이, 일주일 간격으로 체중 증가량을 확인했을 때도, CRE를 처리한 그룹이 그렇지 않은 그룹보다 체중 증가량이 유의하게 감소한 점을 확인하였다.Additionally, as shown in Figure 4b, when the amount of body weight gain was checked at weekly intervals, it was confirmed that the amount of body weight gain was significantly reduced in the group treated with CRE compared to the group not treated with CRE.
또한, 각 그룹별 지방 조직 및 장기의 무게를 확인한 결과, 도 4c에 나타난 바와 같이, CRE 처리 그룹에서 서혜부 백색 지방 조직 (inguinal white adipose tissue 이하 'iWAT') 및 부고환 백색 지방 조직 (epididymal white adipose tissue 이하 'eWAT')의 무게가 감소함을 확인하였으며(상단), 백색 지방 조직(WAT)의 무게를 체중과 대비하여 각 그룹의 비만도를 확인해봤을 때, CRE 처리 그룹이 HFD 그룹 대비 비만도가 감소한 것을 확인하였으며(하단), 이는 상기 조직 및 장기 무게 감소 경향과 일치함을 보였다.In addition, as a result of checking the weight of adipose tissue and organs in each group, as shown in Figure 4c, inguinal white adipose tissue (hereinafter referred to as 'iWAT') and epididymal white adipose tissue in the CRE treatment group It was confirmed that the weight of (hereinafter referred to as 'eWAT') decreased (top), and when the weight of white adipose tissue (WAT) was compared with body weight to check the obesity level of each group, the CRE treatment group showed a decrease in obesity compared to the HFD group. This was confirmed (bottom), and this was consistent with the trend of tissue and organ weight reduction.
또한, CRE가 HFD 유도된 마우스 그룹의 대사 프로필(metabolic profile)을 개선할 수 있는지 상기 실험예 6의 방법으로 확인해본 결과, 도 4d에 나타난 바와 같이, HFD 대비 CRE 처리 그룹에서 TG 및 총 콜레스테롤 함량이 감소함을 확인하였다.In addition, as a result of confirming whether CRE can improve the metabolic profile of the HFD-induced mouse group by the method of Experimental Example 6, as shown in Figure 4d, TG and total cholesterol content in the CRE-treated group compared to the HFD This decrease was confirmed.
또한, 도 4e에 나타난 바와 같이, HFD로 유도된 비만 마우스에서 CRE가 투여된 그룹이 HFD 그룹에 비해 감소된 내장 및 피하 지방 조직을 보이는 것을 확인하였으며, 도 4f 및 도 4g에 나타난 바와 같이, iWAT, eWAT의 지방 조직 모두에서 HFD 그룹에 비해 CRE 처리 그룹이 지방 조직의 크기가 유의하게 감소함을 확인하였다.In addition, as shown in Figure 4e, it was confirmed that the CRE-administered group in HFD-induced obese mice showed reduced visceral and subcutaneous adipose tissue compared to the HFD group, and as shown in Figures 4f and 4g, iWAT , it was confirmed that the size of adipose tissue in the CRE treatment group was significantly reduced compared to the HFD group in all adipose tissues of eWAT.
상기 CRE의 항비만 효과가 생체 내 독성에 기인한 것인지 평가해보기 위하여 상기 실험예 6의 방법으로 혈장 내의 ALP, TP, ALB, T-BIL, D-BIL, BUN, CREA 등을 분석해 본 결과, 표 2에 나타난 바와 같이, CRE를 처리한 그룹과 처리하지 않은 그룹에서의 차이는 나타나지 않았으며 이러한 결과는 CRE는 생체 내 독성을 야기하지 않는 것을 시사한다.In order to evaluate whether the anti-obesity effect of CRE is due to in vivo toxicity, ALP, TP, ALB, T-BIL, D-BIL, BUN, CREA, etc. in plasma were analyzed using the method of Experimental Example 6. Table As shown in Fig. 2, there was no difference between the group treated with CRE and the group not treated with CRE, and these results suggest that CRE does not cause toxicity in vivo.
Figure PCTKR2023019426-appb-img-000002
Figure PCTKR2023019426-appb-img-000002
상기 결과는 CRE가 HFD로 유도된 비만 마우스에서 비정상적인 지방 증가 및 지질함량을 조절하는데 도움이 됨을 시사한다.The above results suggest that CRE helps control abnormal fat gain and lipid content in HFD-induced obese mice.
이상의 실시예를 종합하여, 본 발명자들은 산사나무 뿌리 추출물이 세포 독성 없이 지방 전구 세포 분화를 효과적으로 억제하고, 항산화 효과를 가지며, 지방세포 내 지질 축적을 감소시킬 뿐만 아니라, 지방 세포 분화 및 성숙에 관여하는 전사인자 또는 아디포넥틴의 발현을 억제함을 확인하였고, 고지방 식이 마우스에서 고지방 식이로 인한 간의 지방증 저해 및 간 손상 개선 또는 지방 생성을 억제함을 확인하였는바, 이러한 결과는 산사나무 뿌리 추출물을 지방간 또는 비만의 예방, 치료, 또는 개선을 위한 약제 및 건강기능식품과 이들의 개발을 위한 물질에 이용할 수 있음을 보여준다.Comprehensive of the above examples, the present inventors found that hawthorn root extract effectively inhibits adipocyte differentiation without cytotoxicity, has an antioxidant effect, reduces lipid accumulation in adipocytes, and participates in adipocyte differentiation and maturation. It was confirmed that it suppresses the expression of transcription factors or adiponectin, and it was confirmed that it inhibited liver steatosis and improved liver damage or suppressed fat production caused by a high-fat diet in mice fed a high-fat diet. These results showed that hawthorn root extract was used to treat fatty liver or liver disease. It shows that it can be used as a drug and health functional food for preventing, treating, or improving obesity, and as a substance for their development.
전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술 분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.The description of the present invention described above is for illustrative purposes, 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 features of the present invention. will be. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive.
본 발명자들은 산사나무 뿌리 추출물이 세포 독성 없이 지방 전구 세포 분화를 효과적으로 억제하고, 지방세포 내 지질 축적을 감소시킬 뿐만 아니라, 지방 세포 분화 및 성숙에 관여하는 전사인자의 발현을 억제함을 확인하였다. 또한 고지방 식이 마우스에서 고지방 식이로 인한 간 지방증의 저해 및 간 손상의 개선효과를 확인하였고, 고지방 식이 마우스에서 지방 생성의 억제를 확인하였다. 따라서 본 발명의 산사나무 뿌리 추출물은 지방간 또는 비만의 예방, 치료, 또는 개선용 약학적 조성물 및 건강기능식품 등으로 유용하게 사용될 것으로 기대되는 바 산업상 이용가능성이 있다.The present inventors confirmed that hawthorn root extract effectively inhibits adipocyte differentiation without cytotoxicity, reduces lipid accumulation in adipocytes, and inhibits the expression of transcription factors involved in adipocyte differentiation and maturation. In addition, the effect of inhibiting hepatic steatosis and improving liver damage caused by a high-fat diet was confirmed in high-fat diet mice, and the inhibition of fat production was confirmed in high-fat diet mice. Therefore, the hawthorn root extract of the present invention is expected to be useful in pharmaceutical compositions and health functional foods for preventing, treating, or improving fatty liver disease or obesity, and has industrial applicability.

Claims (31)

  1. 산사나무(Crataegus pinnatifida) 뿌리 추출물을 유효성분으로 포함하는, 지방간의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating fatty liver, comprising hawthorn ( Crataegus pinnatifida ) root extract as an active ingredient.
  2. 제1항에 있어서,According to paragraph 1,
    상기 추출물은 물, 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane), 석유에테르(petroleum ether), 디클로로메탄(dichloromethane), 아임계 유체, 및 초임계 유체로 이루어진 군으로부터 선택된 하나 이상의 용매에 의한 추출물인 것을 특징으로 하는, 지방간의 예방 또는 치료용 약학적 조성물.The extract is water, alcohol with 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, and hexane. Characterized in that it is an extract of fatty liver with one or more solvents selected from the group consisting of (hexane), cyclohexane, petroleum ether, dichloromethane, subcritical fluid, and supercritical fluid. Pharmaceutical composition for prevention or treatment.
  3. 제1항에 있어서,According to paragraph 1,
    상기 지방간은 비알콜성 지방간인, 지방간의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating fatty liver, wherein the fatty liver is non-alcoholic fatty liver.
  4. 제1항에 있어서,According to paragraph 1,
    상기 조성물은 고지방식이로 유도된 간 손상을 개선하는 것을 특징으로 하는, 지방간의 예방 또는 치료용 약학적 조성물.The composition is a pharmaceutical composition for preventing or treating fatty liver, characterized in that it improves liver damage induced by a high-fat diet.
  5. 제1항에 있어서,According to paragraph 1,
    상기 조성물은 고지방식이군의 혈청 ALT 또는 AST 활성을 감소시키는 것을 특징으로 하는, 지방간의 예방 또는 치료용 약학적 조성물.The composition is a pharmaceutical composition for preventing or treating fatty liver, characterized in that it reduces serum ALT or AST activity in the high-fat diet group.
  6. 제1항에 있어서,According to paragraph 1,
    상기 산사나무 뿌리 추출물은 지방세포 분화 및 성숙에 관여하는 전사인자 또는 아디포넥틴의 발현을 억제하는 것을 특징으로 하는, 지방간의 예방 또는 치료용 약학적 조성물.The hawthorn root extract is a pharmaceutical composition for preventing or treating fatty liver, characterized in that it inhibits the expression of adiponectin or transcription factors involved in adipocyte differentiation and maturation.
  7. 제6항에 있어서,According to clause 6,
    상기 전사인자는 퍼옥시좀 증식제 활성화 수용체 감마(PPARγ), 스테롤 조절인자 결합 단백질-1(SREBP-1) 또는 CCAAT-인핸서 결합 단백질 알파(C/EBPα)인 것을 특징으로 하는, 지방간의 예방 또는 치료용 약학적 조성물.Prevention of fatty liver, characterized in that the transcription factor is peroxisome proliferator activated receptor gamma (PPARγ), sterol regulator binding protein-1 (SREBP-1), or CCAAT-enhancer binding protein alpha (C/EBPα), or Pharmaceutical composition for therapeutic use.
  8. 산사나무 뿌리 추출물을 유효성분으로 포함하는, 지방간의 예방 또는 개선용 식품 조성물.A food composition for preventing or improving fatty liver disease, comprising hawthorn root extract as an active ingredient.
  9. 제8항에 있어서,According to clause 8,
    상기 조성물은 건강기능성 식품 조성물인 것을 특징으로 하는, 지방간의 예방 또는 개선용 식품 조성물.The composition is a food composition for preventing or improving fatty liver, characterized in that it is a health functional food composition.
  10. 제8항에 있어서,According to clause 8,
    상기 추출물은 물, 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane), 석유에테르(petroleum ether), 디클로로메탄(dichloromethane), 아임계 유체, 및 초임계 유체로 이루어진 군으로부터 선택된 하나 이상의 용매에 의한 추출물인 것을 특징으로 하는, 지방간의 예방 또는 개선용 식품 조성물.The extract is water, alcohol with 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, and hexane. Characterized in that it is an extract of fatty liver with one or more solvents selected from the group consisting of (hexane), cyclohexane, petroleum ether, dichloromethane, subcritical fluid, and supercritical fluid. Food composition for prevention or improvement.
  11. 제8항에 있어서,According to clause 8,
    상기 지방간은 비알콜성 지방간인, 지방간의 예방 또는 개선용 식품 조성물.A food composition for preventing or improving fatty liver, wherein the fatty liver is non-alcoholic fatty liver.
  12. 제8항에 있어서,According to clause 8,
    상기 조성물은 고지방식이로 유도된 간 손상을 개선하는 것을 특징으로 하는, 지방간의 예방 또는 개선용 식품 조성물.The composition is a food composition for preventing or improving fatty liver, characterized in that it improves liver damage induced by a high-fat diet.
  13. 제8항에 있어서,According to clause 8,
    상기 조성물은 고지방식이군의 혈청 ALT 또는 AST 활성을 감소시키는 것을 특징으로 하는, 지방간의 예방 또는 개선용 식품 조성물.The composition is a food composition for preventing or improving fatty liver, characterized in that it reduces serum ALT or AST activity in the high-fat diet group.
  14. 제8항에 있어서,According to clause 8,
    상기 산사나무 뿌리 추출물은 지방세포 분화 및 성숙에 관여하는 전사인자 또는 아디포넥틴의 발현을 억제하는 것을 특징으로 하는, 지방간의 예방 또는 개선용 식품 조성물.The hawthorn root extract is a food composition for preventing or improving fatty liver, characterized in that it inhibits the expression of adiponectin or transcription factors involved in adipocyte differentiation and maturation.
  15. 제14항에 있어서,According to clause 14,
    상기 전사인자는 퍼옥시좀 증식제 활성화 수용체 감마(PPARγ), 스테롤 조절인자 결합 단백질-1(SREBP-1) 또는 CCAAT-인핸서 결합 단백질 알파(C/EBPα)인 것을 특징으로 하는, 지방간의 예방 또는 개선용 식품 조성물.Prevention of fatty liver, characterized in that the transcription factor is peroxisome proliferator activated receptor gamma (PPARγ), sterol regulator binding protein-1 (SREBP-1), or CCAAT-enhancer binding protein alpha (C/EBPα), or Food composition for improvement.
  16. 산사나무(Crataegus pinnatifida) 뿌리 추출물을 유효성분으로 포함하는, 비만의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating obesity, comprising a hawthorn (Crataegus pinnatifida) root extract as an active ingredient.
  17. 제16항에 있어서,According to clause 16,
    상기 추출물은 물, 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane), 석유에테르(petroleum ether), 디클로로메탄(dichloromethane), 아임계 유체, 및 초임계 유체로 이루어진 군으로부터 선택된 하나 이상의 용매에 의한 추출물인 것을 특징으로 하는, 비만의 예방 또는 치료용 약학적 조성물.The extract is water, alcohol with 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, and hexane. Characterized by being an extract of obesity using one or more solvents selected from the group consisting of (hexane), cyclohexane, petroleum ether, dichloromethane, subcritical fluid, and supercritical fluid. Pharmaceutical composition for prevention or treatment.
  18. 제16항에 있어서,According to clause 16,
    상기 산사나무 뿌리 추출물은 하기 성질중 하나 이상을 가지는 것을 특징으로 하는, 비만의 예방 또는 치료용 약학적 조성물:A pharmaceutical composition for preventing or treating obesity, wherein the hawthorn root extract has one or more of the following properties:
    i) 지방세포의 분화를 억제;i) Inhibit differentiation of adipocytes;
    ii) 세포 내 지질 축적을 감소; 및ii) reduce intracellular lipid accumulation; and
    iii) 지방 세포 분화중 유도되는 산화 스트레스를 감소.iii) Reduces oxidative stress induced during adipocyte differentiation.
  19. 제16항에 있어서,According to clause 16,
    상기 산사나무 뿌리 추출물은 지방세포 분화 및 성숙에 관여하는 전사인자 또는 아디포넥틴의 발현을 억제하는 것을 특징으로 하는, 비만의 예방 또는 치료용 약학적 조성물.The hawthorn root extract is a pharmaceutical composition for preventing or treating obesity, characterized in that it inhibits the expression of adiponectin or transcription factors involved in adipocyte differentiation and maturation.
  20. 제19항에 있어서,According to clause 19,
    상기 전사인자는 퍼옥시좀 증식제 활성화 수용체 감마(PPARγ), 스테롤 조절인자 결합 단백질-1(SREBP-1) 또는 CCAAT-인핸서 결합 단백질 알파(C/EBPα)인 것을 특징으로 하는, 비만의 예방 또는 치료용 약학적 조성물.Prevention of obesity, characterized in that the transcription factor is peroxisome proliferator-activated receptor gamma (PPARγ), sterol regulator binding protein-1 (SREBP-1), or CCAAT-enhancer binding protein alpha (C/EBPα) Pharmaceutical composition for therapeutic use.
  21. 산사나무 뿌리 추출물을 유효성분으로 포함하는, 비만의 예방 또는 개선용 식품 조성물.A food composition for preventing or improving obesity, comprising hawthorn root extract as an active ingredient.
  22. 제21항에 있어서,According to clause 21,
    상기 조성물은 건강기능성 식품 조성물인 것을 특징으로 하는, 비만의 예방 또는 개선용 식품 조성물.The composition is a food composition for preventing or improving obesity, characterized in that it is a health functional food composition.
  23. 제21항에 있어서,According to clause 21,
    상기 추출물은 물, 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane), 석유에테르(petroleum ether), 디클로로메탄(dichloromethane), 아임계 유체, 및 초임계 유체로 이루어진 군으로부터 선택된 하나 이상의 용매에 의한 추출물인 것을 특징으로 하는, 비만의 예방 또는 개선용 식품 조성물.The extract is water, alcohol with 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, and hexane. Characterized by being an extract of obesity using one or more solvents selected from the group consisting of (hexane), cyclohexane, petroleum ether, dichloromethane, subcritical fluid, and supercritical fluid. Food composition for prevention or improvement.
  24. 제21항에 있어서,According to clause 21,
    상기 산사나무 뿌리 추출물은 하기 성질중 하나 이상을 가지는 것을 특징으로 하는, 비만의 예방 또는 개선용 식품 조성물:A food composition for preventing or improving obesity, wherein the hawthorn root extract has one or more of the following properties:
    i) 지방세포의 분화를 억제;i) Inhibit differentiation of adipocytes;
    ii) 세포 내 지질 축적을 감소; 및ii) reduce intracellular lipid accumulation; and
    iii) 지방 세포 분화중 유도되는 산화 스트레스를 감소.iii) Reduces oxidative stress induced during adipocyte differentiation.
  25. 제21항에 있어서,According to clause 21,
    상기 산사나무 뿌리 추출물은 지방세포 분화 및 성숙에 관여하는 전사인자 또는 아디포넥틴의 발현을 억제하는 것을 특징으로 하는, 비만의 예방 또는 개선용 식품 조성물.A food composition for preventing or improving obesity, wherein the hawthorn root extract inhibits the expression of adiponectin or transcription factors involved in adipocyte differentiation and maturation.
  26. 제25항에 있어서,According to clause 25,
    상기 전사인자는 퍼옥시좀 증식제 활성화 수용체 감마(PPARγ), 스테롤 조절인자 결합 단백질-1(SREBP-1) 또는 CCAAT-인핸서 결합 단백질 알파(C/EBPα)인 것을 특징으로 하는, 비만의 예방 또는 개선용 식품 조성물.Prevention of obesity, characterized in that the transcription factor is peroxisome proliferator-activated receptor gamma (PPARγ), sterol regulator binding protein-1 (SREBP-1), or CCAAT-enhancer binding protein alpha (C/EBPα) Food composition for improvement.
  27. 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 화장료 조성물.A cosmetic composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  28. 산사나무 뿌리 추출물을 유효성분으로 포함하는 비만의 예방 또는 개선용 의약외품 조성물.A quasi-drug composition for preventing or improving obesity containing hawthorn root extract as an active ingredient.
  29. 산사나무(Crataegus pinnatifida) 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물을 이를 필요로 하는 개체에게 투여하는 단계를 포함하는, 지방간 또는 비만의 예방, 개선 또는 치료 방법.A method for preventing, improving or treating fatty liver or obesity, comprising administering a hawthorn ( Crataegus pinnatifida ) root extract or a composition containing the same as an active ingredient to an individual in need thereof.
  30. 산사나무(Crataegus pinnatifida) 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 지방간 또는 비만의 예방, 개선 또는 치료 용도.Use of hawthorn ( Crataegus pinnatifida ) root extract or a composition containing it as an active ingredient to prevent, improve, or treat fatty liver or obesity.
  31. 산사나무(Crataegus pinnatifida) 뿌리 추출물 또는 이를 유효성분으로 포함하는 조성물의 지방간 또는 비만의 예방, 개선 또는 치료 제제를 제조하기 위한 용도.Use of hawthorn ( Crataegus pinnatifida ) root extract or a composition containing the same as an active ingredient for preparing a preparation for preventing, improving, or treating fatty liver disease or obesity.
PCT/KR2023/019426 2022-12-01 2023-11-29 Pharmaceutical composition for preventing or treating fatty liver or obesity, comprising extract of crataegus pinnatifida bunge roots as active ingredient WO2024117763A1 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR10-2022-0165566 2022-12-01
KR1020220165578A KR20240081878A (en) 2022-12-01 2022-12-01 Pharmaceutical Composition comprising Crataegus pinnatifida root extract as an active ingredient for Preventing or Treating obesity
KR1020220165566A KR20240081872A (en) 2022-12-01 2022-12-01 Pharmaceutical Composition comprising Crataegus pinnatifida root extract as an active ingredient for Preventing or Treating fatty liver
KR10-2022-0165578 2022-12-01

Publications (1)

Publication Number Publication Date
WO2024117763A1 true WO2024117763A1 (en) 2024-06-06

Family

ID=91324687

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2023/019426 WO2024117763A1 (en) 2022-12-01 2023-11-29 Pharmaceutical composition for preventing or treating fatty liver or obesity, comprising extract of crataegus pinnatifida bunge roots as active ingredient

Country Status (1)

Country Link
WO (1) WO2024117763A1 (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20130075036A (en) * 2011-12-27 2013-07-05 이강수 Food composition for anti-obesity and preparation thereof
JP2014208620A (en) * 2013-03-25 2014-11-06 株式会社漢方医科学研究所 Anti-obesity agent
KR20160070872A (en) * 2014-12-10 2016-06-21 원광대학교산학협력단 Composition for prevention or treatment of nonalcoholic fatty liver disease comprising Crataegi Fructus extract
KR20180114395A (en) * 2017-04-10 2018-10-18 주식회사 엘지생활건강 A health functional food comprising composite material extract for preventing or treating obesity
KR20210030192A (en) * 2019-09-09 2021-03-17 (주)리즈바이오텍 Pharmaceutical composition containing complex Rubus crataegifolius, Crataegus pinnatifida Bunge, Cinnamomum cassia extracts for prevention or treatment of metabolic syndrome

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20130075036A (en) * 2011-12-27 2013-07-05 이강수 Food composition for anti-obesity and preparation thereof
JP2014208620A (en) * 2013-03-25 2014-11-06 株式会社漢方医科学研究所 Anti-obesity agent
KR20160070872A (en) * 2014-12-10 2016-06-21 원광대학교산학협력단 Composition for prevention or treatment of nonalcoholic fatty liver disease comprising Crataegi Fructus extract
KR20180114395A (en) * 2017-04-10 2018-10-18 주식회사 엘지생활건강 A health functional food comprising composite material extract for preventing or treating obesity
KR20210030192A (en) * 2019-09-09 2021-03-17 (주)리즈바이오텍 Pharmaceutical composition containing complex Rubus crataegifolius, Crataegus pinnatifida Bunge, Cinnamomum cassia extracts for prevention or treatment of metabolic syndrome

Similar Documents

Publication Publication Date Title
WO2017039365A1 (en) Method for inhibiting absorption of and/or promoting excretion of lipids using d-psicose
WO2012124888A2 (en) Composition comprising the extract of herbal combination for preventing or treating diabetic peripheral neuropathy
WO2016032249A1 (en) Pharmaceutical composition containing vaccinium bracteatum thunb. extract or fraction thereof as active ingredient for preventing or treating neuroinflammation or neuro-degenerative diseases
WO2020218720A1 (en) Composition for preventing or treating muscular disorders or improving muscular functions, containing leonurus japonicus extract or leonurine
WO2010087577A2 (en) Use of thymus capitatus extract, satureja hortensis extract, or carvacrol for treating metabolic diseases
WO2015111832A1 (en) Composition for preventing or treating prostate-related diseases, containing poncirus trifoliate extract
WO2015030336A1 (en) Composition containing composite extract of grape and schisandra chinensis for preventing or treating metabolic syndrome-related diseases
WO2019017677A2 (en) Composition comprising citral as effective ingredient for exhibiting effect of muscle strengthening, muscle enhancement, muscle differentiation, muscle regeneration, or sarcopenia suppression
WO2013069934A1 (en) Composition for treating and preventing obesity, containing wheatgrass extract as active ingredient
WO2024117763A1 (en) Pharmaceutical composition for preventing or treating fatty liver or obesity, comprising extract of crataegus pinnatifida bunge roots as active ingredient
WO2013012117A1 (en) Pharmaceutical compositions for preventing or treating inflammatory diseases, comprising phytosterol compound
WO2010041908A2 (en) Novel use of panduratin derivative or boesenbergia pandurata extract
WO2021002642A1 (en) Composition for preventing or treating rheumatoid arthritis, comprising snake venom
WO2020204479A1 (en) Composition for alleviating venous circulation disorders comprising grape leaf extract and centella asiatica extract
WO2015009047A1 (en) Composition containing humulus japonicus extract for preventing or treating insulin resistance related diseases
WO2010143825A2 (en) Anti-arthritic agent using cyathula officinalis
WO2015152653A1 (en) Composition comprising extract of alpine wormwood
WO2015105373A1 (en) Composition for prevention or treatment of asthma, comprising e uonymus alatus extract or fraction thereof
WO2014014177A1 (en) Composition comprising dendropanax morbifera extract or compound derived therefrom as active ingredient for preventing and treating benign prostatic hyperplasia
WO2022265281A1 (en) Composition for preventing, improving, or treating degenerative arthritis comprising steamed ginger extract or 1-dehydro-6-gingerdione isolated therefrom as active ingredient
WO2018190638A1 (en) Composition for preventing or treating corneal diseases, containing glycine max extract
WO2023017958A1 (en) Composition for preventing, treating, or alleviating arthritis, comprising elaeocarpus sylvestris extract or mixture of elaeocarpus sylvestris extract and sulfasalazine as an active ingredient
WO2011122805A2 (en) A composition comprising ajoene for preventing or treating a disease caused by overexpression of lxr-alpha
WO2022270760A1 (en) Method for treating non-alcoholic steatohepatitis through co-administration of curcumin derivative and tgf-β receptor inhibitor
WO2023182757A1 (en) Composition for preventing and treating stroke