WO2013022178A1 - Composition pour prévention et traitement de l'obésité contenant un principe actif sous la forme d'un extrait de préparation végétale mélangée à base de lonicera japonica et d'aurantii nobilis pericarpium fermenté ou non fermenté, et son utilisation - Google Patents

Composition pour prévention et traitement de l'obésité contenant un principe actif sous la forme d'un extrait de préparation végétale mélangée à base de lonicera japonica et d'aurantii nobilis pericarpium fermenté ou non fermenté, et son utilisation Download PDF

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WO2013022178A1
WO2013022178A1 PCT/KR2012/004237 KR2012004237W WO2013022178A1 WO 2013022178 A1 WO2013022178 A1 WO 2013022178A1 KR 2012004237 W KR2012004237 W KR 2012004237W WO 2013022178 A1 WO2013022178 A1 WO 2013022178A1
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fermented
extract
obesity
fermentation
silver
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English (en)
Korean (ko)
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김미려
이은실
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대구한의대학교 산학협력단
영농조합법인 이도
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Publication of WO2013022178A1 publication Critical patent/WO2013022178A1/fr

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    • 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)
    • A61K36/35—Caprifoliaceae (Honeysuckle family)
    • 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)
    • A61K36/75—Rutaceae (Rue family)
    • A61K36/752—Citrus, e.g. lime, orange or lemon
    • 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)
    • A61K36/35—Caprifoliaceae (Honeysuckle family)
    • A61K36/355—Lonicera (honeysuckle)
    • 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
    • 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

Definitions

  • the present invention relates to a composition for the prevention and treatment of obesity and its use containing fermented or unfermented sterling silver and dermis mixed herbal extracts as an active ingredient.
  • Cinnamaldehyde prevents adipocyte differentiation and adipogenesis via regulation of peroxisome proliferator-activated receptor- ⁇ (PPAR ⁇ ) and AMP-activated protein kinase (AMPK) pathways, J Agric Food Chem, Apr 27; 59; (8) , pp. 3666-73, 2011
  • PPAR ⁇ peroxisome proliferator-activated receptor- ⁇
  • AMPK AMP-activated protein kinase
  • Obesity is a phenomenon caused by an imbalance between the intake and consumption of energy, which means an excess of fat in the body. Although it is not known yet, because it is caused by various causes, it is generally recognized to be caused by excessive calorie intake, endocrine disorders, lack of exercise, and genetic factors. It is also commonly referred to as lifestyle disease because it is closely related to lifestyle. Since 2000, the number of people with metabolic syndrome, which is susceptible to chronic diseases such as heart disease and diabetes, has increased 65% in recent five years. As a result, various problems such as suffering and poor quality of life of the patient as well as excessive medical expenses and reduced productivity are socially occurring.
  • Metabolic disease is estimated to increase by about 50% to about 1.5 billion people.
  • Obesity refers to a condition in which the body's fat is excessively accumulated to damage health, and can be defined as obesity when there is an increase of 20% or more of the individual's standard weight. Specifically, the weight of fat accounts for more than 25% of body fat in men and more than 30% in women (Van der Ploeg LH.Related Obesity: an epidemic in need of therapeutics.Curr Opin Chem Biol 4 (4) , pp. 452-460, 2004). All inde trend worldwide increase in overweight or obese people that has been a significant increase in the incidence of obesity in our country recently (Kim DM, et al., Prevalence of obesity in Korea. Obes Rev. 6, pp.117-121 , 2005).
  • Obesity is associated with the prevalence of various degenerative diseases, such as diabetes, hypertension, cardiovascular disease, joint disease, lung disease, and some cancers, because excess energy is converted to fat due to an imbalance between energy intake and consumption.
  • the financial burden and loss of life incurred are enormous (Daily JW, Cha YS. Cellutrient intake and obesity. J Food Sci Nutr. 5 (1) , pp.58-64, 2000).
  • obesity management is more important because the body mass index is at least abdominal obesity compared with Westerners, and the sensitivity of arterial diseases such as hypertension, diabetes, and hyperlipidemia is high.
  • the medical community recognizes the appetite suppressant sibutramine and the fat absorption inhibitor olistat as obesity drugs that can be used for a long time, but there are still questions about side effects, so diet, exercise and behavior therapy are used to safely lose weight. Including a multifaceted approach is recommended.
  • Lonicerae Flos is a perennial herbaceous vine that belongs to the Caprifoliaceae family. It is harvested before flowering in summer and dried in shade. Tannin, inositol, sterol, chlorogenic acid and isochlogenic acid have been reported as ingredients, and flavonoids include luteolin, apigenin and luteolin-7-O-rhamnoglucoside. Contains lonicerin, inositol, essential oils and quercetin. It is sweet and weak in coldness and detoxification, so it is an important medicine to cure heat venom (Jungkuk University of Medicine, College of Medicine, Herbal Medicine, Younglimsa, pp.242-244 , 2006).
  • the dermis (Citri Reticullatae Pericarpium) is a dry-married fruit peel of the mature fruit of the evergreen subfamily, Tangerine, and the same genus, which belongs to the Unhyang family. It is harvested in November and dried in the sun. Its components include hesperidin, naringin poncirin, nobilein, neoesprin, nobileting, neohesprindin and tanderetin as flavonoids.
  • the taste is spicy and bitter, the medicinal properties are warm, fragrant, and mainly enter into the rain and menopause, which is effective.
  • the spicy taste is divergent and hangs, and the taste is bitter and warm.
  • Fermented herbal medicine refers to herbal medicines fermented using purely isolated microorganisms such as microorganisms in the air or lactic acid bacteria after steaming or boiling traditional herbal medicines so that microorganisms can use them well through traditional fermentation methods. It is a kind of processing method that maximizes the body absorption rate and bioavailability of Chinese herbal medicines and improves the pharmacological function as well as the formulation improvement and preparation method of Chinese medicine, thereby creating new demand for Chinese medicine and producing new high value-added Chinese medicine products. It is meant to be able to develop.
  • various manufacturing methods and fermented herbal medicines are being developed, and fermented red ginseng products and fermented herbal cosmetics can be considered as representative products.
  • the mixed herbal extract obtained by fermenting sterling silver and dermis can be used as a composition for preventing and treating obesity.
  • the present inventors have conducted research to develop an effective therapeutic agent for obesity, inhibiting the differentiation of 3T3-L1, which is a fat cell, from gold and silver dermis fermented extracts, and reducing the body weight, body fat, and blood lipid levels in obesity-induced animals using high-fat diet.
  • 3T3-L1 is a fat cell
  • adiponectin adiponectin
  • the present invention provides a pharmaceutical composition for the prevention and treatment of obesity containing fermented or unfermented gold and silver dermis mixed herbal extract as an active ingredient.
  • the present invention also provides a health functional food for the prevention and improvement of obesity, containing fermented or unfermented sterling silver and dermis mixed herbal extracts as an active ingredient.
  • Mixed herbal extracts as defined herein include extracts soluble in water, preferably lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, preferably extracts soluble in water.
  • Preferred weight blending ratios of the mixed herbals as defined herein include blends in which the weight blending ratio (w / w) of gold to silver dermis is 1-5: 1, more preferably 1-3: 1 (w / w). It is characterized by that.
  • the fermentation extracts defined herein are sterilized from 10 to 70 minutes, preferably from 20 to 30 minutes, at 80 to 150 ° C. for samples of dried individual herbs with a water content of less than 20%, preferably less than 10%. Sterilization step; Sterilized samples can be fermented for Aspergillus oryzae, Aspergillus niger, Lactobacillus acidophilus Lactobacillus casei, and other food processing.
  • a strain preferably Aspergillus oryzae, is added in an amount of 1 to 30%, preferably 1 to 20% of the weight of the sample so that 10 to 50%, preferably 20 to 40%
  • the selected fermented product is a process comprising a fifth step of performing a process of quenching for 5 minutes to 60 minutes, preferably 10 minutes to 30 minutes with far-infrared herb at 100 to 300 °C, preferably 150 to 200 °C Obtainable.
  • the mixed herbal extract of the present invention can be obtained as follows.
  • Fermented or unfermented mixed herbal extract of the present invention first, the gold and silver or dermis individual herbal medicines, the water moisture content of less than 20%, preferably less than 10% of the sample dried at 80 to 150 °C 10 minutes to 70 minutes, preferably The sterilization step of undergoing a sterilization process for 20 to 30 minutes; Sterilized samples can be fermented for Aspergillus oryzae, Aspergillus niger, Lactobacillus acidophilus Lactobacillus casei, and other food processing. The strain is added in an amount of 1 to 30%, preferably 1 to 20%, of the sample weight, so that the temperature is 10 to 50 ° C., preferably 20 to 40 ° C. under conditions of 10 to 50%, preferably 20 to 40%, moisture.
  • the selected fermented product is subjected to a fifth step of carrying out the process of sieving for 5 minutes to 60 minutes, preferably 10 minutes to 30 minutes with far-infrared herb at 100 to 300 ° C, preferably 150 to 200 ° C. Fermentation samples of can be obtained.
  • the fermentation sample obtained in the above step or the unfermented dry medicinal herb gold and silver dermis are washed, and the weight mixing ratio (w / w) of gold to silver dermis is 1-5: 1, more preferably 1-3: 1 (w / w) and 1 to 20 times (w / v) weight of the sample, preferably 1 to 10 times (w / v) weight of water, methanol, ethanol or a mixed solvent thereof, preferably 50 to water
  • the extract obtained after performing hot water extraction, cold needle extraction or ultrasonic extraction, preferably hot water extraction at 120 ° C., preferably about 80-100 ° C. for 1 hour to 5 hours, preferably 2 hours to 4 hours, is filtered with a filter paper.
  • the filtrate obtained after the filtration may be lyophilized, room temperature or hot air dried, preferably lyophilized, to obtain fermented or unfermented mixed herbal extracts composed of gold and silver, the dermis of the present invention.
  • the present invention provides a pharmaceutical composition for the treatment and prevention of obesity, containing the fermented or unfermented sterling silver and dermis herbal extract obtained by the manufacturing method and the manufacturing process as an active ingredient.
  • the present invention also provides a method for treating obese patients, comprising administering a mixed herbal extract of fermented or unfermented sterling silver and dermis to an obese patient.
  • the present invention also provides the use of a mixed herbal extract of fermented or unfermented sterling silver and dermis for the manufacture of a medicament for the treatment of obesity.
  • the gold and silver dermis herbal extracts obtained above reduce fat differentiation of adipocytes, and show weight loss, body fat reduction and blood lipid concentration in rats induced obesity using high fat diet.
  • the pharmaceutical composition for treating obesity of the present invention comprises 0.01 to 80% by weight of the extract, preferably 10 to 50% by weight, based on the total weight of the composition.
  • compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
  • compositions of the extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other pharmaceutically active compounds, as well as in any suitable collection.
  • the pharmaceutical composition comprising the extract according to the present invention is formulated in the form of external preparations, suppositories, and sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc. Can be used.
  • Carriers, excipients and diluents that may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, and the like in the extracts and fractions. (sucrose), lactose (lactose), gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.
  • Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin.
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
  • the non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.
  • As the base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol gelatin and the like can be used.
  • the preferred dosage of the pharmaceutical composition comprising the extract of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the pharmaceutical composition comprising the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
  • Fermented or unfermented sterling silver and dermis extracts of the present invention can be administered to mammals such as mice, mice, livestock, humans and the like by a variety of routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or Intracerebroventricular injection.
  • the present invention provides a health functional food for the prevention and improvement of obesity, containing fermented or unfermented sterling silver and dermis mixed herbal extracts having an excellent inhibitory effect on obesity as an active ingredient.
  • health functional food means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. 6767, and “functional” means It means ingestion for the purpose of obtaining useful effects on health use such as nutrient control or physiological action on structure and function.
  • the extract comprises 0.01 to 95% by weight, preferably 1 to 80% by weight relative to the total weight of the composition.
  • a pharmaceutical dosage form such as powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, or health functional foods in the form of tea bags, leach teas, and health drinks. Processing is possible.
  • the present invention also provides a dietary supplement containing as a main ingredient a mixed herbal extract of fermented or non-fermented gold and silver and dermis exhibiting anti-obesity activity.
  • the health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the extract as an essential ingredient in the indicated proportions, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks.
  • natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents such as, tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
  • the proportion of the natural carbohydrate is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.
  • the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
  • the compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
  • the extract of the present invention may be added to food or beverage for the purpose of preventing obesity.
  • the amount of the extract in the food or beverage can be added in 0.01 to 15% by weight of the total food weight
  • the health beverage composition is added in a ratio of 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml Can be.
  • Fermented or unfermented mixed gold and silver extracts of the present invention are effective in inhibiting fat differentiation in mouse embryo fibroblast-derived adipocytes, 3T3-L1, and in obese rats induced by high fat diet.
  • body fat reduction and blood lipid concentrations, leptin and adiponectin reduce blood levels, which can be useful in pharmaceutical compositions, dietary supplements, and health supplements for the prevention and treatment of obesity. Can be.
  • 1 is a diagram showing the effect of F-CL on MTT assay of 3T3-L1, a cell line of adipocytes,
  • FIG. 2 is a diagram showing the effect of F-CL on Oil Red-O staining of 3T3-L1, a cell line of adipocytes,
  • FIG. 3 is a diagram showing the effect of CL and F-CL on the effect of the herbal medicine extract on AMPK and ACC activity
  • Figure 4 is a diagram showing the effect of F-CL on the weight of obese mice due to high-fat diet administration
  • FIG. 5 is a diagram showing the effect of F-CL on the liver weight of obese mice due to high-fat diet administration
  • Figure 6 (a-d) is a diagram showing the effect of F-CL on the body fat weight of obese mice due to high-fat diet administration
  • Figure 7 (a-c) is a diagram showing the effect of F-CL on the blood lipid concentration of obese mice due to high-fat diet administration
  • Figure 8 (d-f) is a diagram showing the effect of F-CL on the blood lipid concentration of obese mice due to high-fat diet administration
  • FIG. 9 is a diagram showing the effect of F-CL on the concentration of Adiponectin, Leptin in the blood of obese rats due to high-fat diet administration,
  • FIG. 10 is a diagram showing the effect of F-CL on AMPK and ACC activity in the tissues of obese mice due to high-fat diet administration.
  • the above ingredients are mixed and filled in an airtight cloth to prepare a powder.
  • tablets are prepared by tableting according to a conventional method for preparing tablets.
  • the above ingredients are mixed and filled into gelatin capsules to prepare capsules.
  • the amount of the above ingredient is prepared per ampoule (2 ml).
  • each component is added to the purified water to dissolve it, the lemon flavor is added appropriately, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled in a brown bottle.
  • the solution is prepared by sterilization.
  • Vitamin B6 0.5 mg
  • composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method.
  • the granules may be prepared and used for preparing a health food composition according to a conventional method.
  • composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment
  • compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.
  • Fermented gold silver, dermis, unfermented gold silver and dermis fermented by the above process was measured 5 kg in a 3: 2 weight ratio, and then distilled water of 10 times and then heated to 95 °C and extracted for 12 hours. After extraction, the precipitate was filtered using a filter paper (Advantec, 71101903), and the filtrate was concentrated under reduced pressure in a rotary vacuum distillation machine (EYELA, A-1000s). The concentrate was left for 12 hours in a -70 ° C. deep freezer and dried for 72 hours in a freeze dryer for 2.1 kg (yield 42%) of fermented mixed extract powder and the fermentation process was not performed. 1.6 kg (yield 32% yield) of the non-fermented mixed extract powder prepared from the material was obtained (hereinafter referred to as 'F-CL' and the fermented mixed extract 'CL').
  • 3T3-L1 mouse-derived embryonic fibroblasts were purchased from the American Type Culture Collection (ATCC, CL-173TM), Dulbeccos modified Eagles media (DMEM, SH30243.01), Bovine calf serum (BCS, SH30401.01) , Fetal bovine serum (FBS, SH30396.03) and penicillin and streptomycin (PS) were purchased from Hyclone (SV30010, Hyclone, USA).
  • Insulin (090-03446), dimethyl sulfoxide (DMSO, D2650), Oil Red-O (O0625-25), dexamethasone (DEX, D4902-100MG) and isobutylmethyl Xanthine (isobutylmethylxanthine (IBMX, I5879-250MG) was purchased from Sigma-Aldrich company, St. Louis, Mo., USA and used.
  • mice 5 weeks old ICR mice (Orient Corp.) weighing 28-32 g in weight, and were fed in the animal breeding room of Daegu Haany University under constant conditions (temperature: 21 ⁇ 2, contrast: 12 hour contrast cycle). Free intake of negative water was allowed, and sufficient water and food were provided until the start of the experiment.
  • 3T3-L1 mouse embryo fibroblasts were cultured by the following experimental method (Lin. J. et al., Green Tea Polyphenol Epigallocatechin Gallate Inhibits Adipogenesis and Induces Apoptosis in 3T3-L1 Adipocytes, Obesity Research, 13 (6) , pp. 982-990, 2005).
  • DMEM Dulbecco's modified Eagle's medium
  • DMEM bovine calf serum
  • FBS fetal bovine serum
  • a pellet-type solid feed (# 102038, AIN-76A Based High Fat / High Carb Purified) containing 1% cholesterol and 15% Lard was added to the diet based on the AIN-76 composition.
  • Rodent Diet with Cholesterol & Cholic Acid, Dyet, USA was fed for 6 weeks and 15 animals were assigned to each group. That is, divided into normal group (Normal), high fat control group (HF), high fat feed unfermented mixed extract (CL-5%), high fat feed fermented mixed extract (F-CL-5%), and the sample group in Example 1
  • the lyophilized mixed herbal extract powder obtained in was added to the high fat diet by 5% and fed with water every morning, and 20 g of feed were allocated daily.
  • MTT assay methylthiazol tetrazolium bromide
  • the MTT assay is a method of measuring the absorbance by dissolving blue formazan crystals produced by dehydrogenase in mitochondria of living cells with DMSO (dimethyl sulfoxide).
  • DMSO dimethyl sulfoxide
  • the 3T3-L1 cell line cultured in Reference Example 1 was used, and the cells in exponential growth phase were used for the experiment. In other words, 5 ⁇ 10 4 cells / ml was mixed well with the same amount of 5% tryphan without any treatment for a certain period of time, and the number of living cells was measured.
  • the viability test was evaluated using the average value of all three wells. Add 50 ⁇ l of MTT solution (2 mg / ml) per well and react for 2 hours at 37 ° C and 5% CO 2 incubator. Remove the medium and add 100 ⁇ l of DMSO per well for 5 minutes. After dissolution, absorbance was measured at 540 nm of ELISA reader (Techan, Germany).
  • fermented herbal medicine (F-CL) and unfermented herbal medicine (CL) were treated for 24 hours with different concentrations (0.1, 0.3, 1 mg / ml), and the MTT assay was tested.
  • concentrations 0.1, 0.3, 1 mg / ml
  • MTT assay was tested.
  • the survival rate of the adipocytes was observed to be decreased, and cell viability was significantly inhibited than the fermented medicinal herb extract, especially at 0.1 mg / ml of fermented medicinal herbs (F-CL).
  • Oil Red-O staining is a method of measuring the amount of intracellular fat produced by staining differentiated 3T3-L1 cells with Oil Red-O reagent.
  • the medium of the differentiated cells was removed, the cells were washed with D-PBS (DPBS, SH30028.03), fixed with pH 7.2 Cacodylate buffer for 2 hours, and stained with Oil-red O Solution. After the cell staining was washed three times with 40% Isopropyl alcohol (2-Propanol, 67-63-0) and dried to observe the size of the fat in the cell under an optical microscope (AE 31, Motic, USA).
  • the fermented and unfermented mixed extracts were treated with 100 ⁇ g / ml (B), 150 ⁇ g / ml (C), and 200 ⁇ g / ml (D), respectively, during differentiation of 3T3-L1 cells.
  • B 100 ⁇ g / ml
  • C 150 ⁇ g / ml
  • D 200 ⁇ g / ml
  • 3T3-L1 Cell Western was performed to investigate the effects of fermented extracts on AMPK, ACC and PPAR- ⁇ activity (Huang B. et, al, Cinnamaldehyde prevents adipocyte differentiation and adipogenesis via regulation of peroxisome proliferator-activated receptor- ⁇ (PPAR ⁇ ) and AMP-activated protein kinase (AMPK) pathways, J Agric Food Chem, Apr 27; 59 (8), pp. 3666-73, 2011).
  • PPAR ⁇ peroxisome proliferator-activated receptor- ⁇
  • AMPK AMP-activated protein kinase
  • FER body weight gain / food intake
  • liver and site-specific visceral adipose tissue epididymal, visceral adipose tissue
  • brown adipose tissue BAT
  • the liver weight, epididymal fat weight, peripheral fat weight, visceral weight, and brown fat were significantly increased in the high fat diet group (HF) group compared to the normal group.
  • HF high fat diet group
  • CL and F-CL groups hepatic weight, epididymal fat weight, peri-dial adipose tissue weight, and brown fat weight were decreased, especially in the F-CL group.
  • the triglyceride content was determined by Van Handel et al. (Van Handel E. et al, Micromethod for the direct determination of serum triglycerides, J. Lab. Clin. Med. 50 (1) , pp.152-157, 1957). Test reagents (L-type TG M, Wako, Japan) were used. High-density lipoprotein (HDL-cholesterol) and low-density lipoprotein (LDL-cholesterol) concentrations were determined by Warnick's enzyme method (Warnick GR et al., Dextran sulfate-Mg 2+ precipitation procedure for quantitation of high density lipoprotein cholesterol, Clin. Chem. 20 (6) , pp.
  • ALT, AST and triglyceride concentrations and low density lipoprotein (LDL) content in serum were increased in the high fat diet group (HF) group compared to the normal group.
  • ALT, AST and triglyceride concentrations and low density lipoprotein (LDL) content in serum were decreased by administration of F-CL, and high density lipoprotein (HDL) content in serum was decreased in high fat diet group (HF) group compared to normal group.
  • F-CL administration tended to increase HDL content.
  • the serum content of adiponectin secreted from adipocytes while acting as an insulin-sensitive hormone was increased in the normal group (NOR) than the high-fat diet group (HF), by F-CL administration.
  • the adiponectin content in the serum tended to be higher than that in the high fat diet group (HF).
  • HF high fat diet administered group
  • NOR normal group
  • 3T3-L1 Cell Western was performed to investigate the effects of fermented extracts on AMPK, ACC and PPAR- ⁇ activity (Murase T. et, al, Nootkatone, a characteristic constituent of grapefruit, stimulates energy metabolism and prevents diet- induced obesity by activating AMPK, Am J Physiol Endocrinol Metab, 299 (2), pp. E266-75, 2010).
  • Tissue was used with RIPA buffer (150 mM NaCl, 0.5% Triton X-100, 50 mM Tris-HCl, pH 7.4, 25 mM NaF, 20 mM EGTA, 1 mM DTT, 1 mM Na 3 VO 4 , protease inhibitor cocktail) After homogenization, the supernatant was collected by centrifugation at 14,000 g for 10 minutes at 4 ° C., the protein content was quantified with BCA reagent, and 20 ⁇ g was electrophoresed on 8% SDS-PAGE. The developed gel was transferred to PVDF membrane (Millipore) and blocked with 5% bovine serum albumin (BSA) -TBST (TBS, 0.1% Tween-20).
  • BSA bovine serum albumin
  • Fermented or unfermented mixed gold and silver extracts of the present invention are effective in inhibiting fat differentiation in mouse embryo fibroblast-derived adipocytes, 3T3-L1, and in obese rats induced by high fat diet.
  • body fat reduction and blood lipid concentrations, leptin and adiponectin reduce blood levels, which can be useful in pharmaceutical compositions, dietary supplements, and health supplements for the prevention and treatment of obesity. Can be.

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Abstract

La présente invention concerne une composition qui permet de prévenir et de traiter l'obésité et qui contient un principe actif sous la forme d'un extrait de préparation végétale mélangée à base de Lonicera Japonica et d'Aurantii Nobilis Pericarpium fermenté ou non fermenté. L'extrait de préparation végétale de la présente invention peut être utilisé de manière avantageuse dans des produits pharmaceutiques ou dans des aliments sains et fonctionnels pour la prévention et le traitement de l'obésité, étant donné que l'extrait de la présente invention est efficace dans la suppression de la différenciation adipeuse de cellules 3T3-L1 qui sont des adipocytes dérivés des souris, et, chez des rats blancs chez lesquels l'obésité a été induite à l'aide d'un régime à forte teneur lipidique, l'extrait de la présente invention est efficace dans la réduction du poids corporel et des graisses corporelles, ainsi que dans la réduction de la concentration de lipides dans le sang.
PCT/KR2012/004237 2011-08-10 2012-05-30 Composition pour prévention et traitement de l'obésité contenant un principe actif sous la forme d'un extrait de préparation végétale mélangée à base de lonicera japonica et d'aurantii nobilis pericarpium fermenté ou non fermenté, et son utilisation WO2013022178A1 (fr)

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KR10-2011-0079661 2011-08-10
KR1020110079661A KR101344189B1 (ko) 2011-08-10 2011-08-10 발효 또는 미발효된 금은화 및 진피 혼합생약 추출물을 유효성분으로 함유하는 비만증의 예방 및 치료용 조성물

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KR102087041B1 (ko) * 2019-07-08 2020-03-10 (주)앤이바이오플러스 다이어트를 위한 건강식품 조성물의 제조방법

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108478766A (zh) * 2018-05-22 2018-09-04 北京未医堂健康科技有限公司 一种治疗体内湿气和微循环淤堵的药物及其制备方法

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