KR20150071282A - Pharmaceutical or food composition having TGR5 agonist activity - Google Patents

Abstract

The present invention relates to a pharmaceutical composition or a food composition which is safe due to few side effects, and has excellent TGR5 agonist activity comprising one or more extracts selected from a group consisting of Erigeron annuus, Sasa borealis(Hack.) Makino, cypress, ricinus, sunberry, Stellaria alsine var. undulata Ohwi, angelica, and zanthoxylum piperitum seed as an active ingredient. The composition of the present invention can be used as a pharmaceutical composition or a food composition for treating or preventing diabetes; treating or preventingobesity; protecrting nerve cells; controlling digestion and absorption; or alleviating or treating liver diseases or heart diseases.

Description

A pharmaceutical composition or food composition having TGR5 agonist activity,

The present invention relates to a pharmaceutical composition or a food composition. More specifically, the present invention relates to a pharmaceutical composition or a food composition comprising a plant extract that acts as agonists in combination with TGR5 present in the cell membrane of various tissues in the body as a specific disease or disease To a pharmaceutical composition or food composition useful in the treatment of cancer.

The G protein-coupled receptor 5 (TGR5, GPBAR1, GPR131) is a class A G protein that is present on the cell membrane of the liver, skeletal muscle, intestine, brown adipose tissue, -protein-coupled receptor (GPCR) (1. AT Londregan, DW Piotrowski, K. Futatsugi, JS Warmus et al., Discovery of 5-phenoxy-1,3-dimethyl-1H- pyrazole-4-carboxamides as potent agonists of TGR5 via sequential combinatorial libraries. Bioorganic & Medicinal Chemistry Letters 23 (2013) 1407-1411).

Cholanic acid and bile acid derivatives bind to TGR5 to act as TGR5 agonists. When TGR5 is activated, it increases intracellular cAMP levels and increases protein kinase A (PKA) activity. This activation of PKA plays an important role in the regulation of metabolism, glucose and energy homeostasis, specifically in tissues in which TGR5 is present. In contrast, activation of TGR5, which is present in intestinal endocrine cells, increases secretion of glucagon-like peptide-1 (GLP-1) from intestinal endocrine cells (Katsuma, S., Hirasawa, A., Tsujimoto, G. Biochem (TGF-β1), which is an endothelial nitric oxide synthase (eNOS) and nitric oxide (TGF-β) in liver cells (Kietel, V., Reinehr, R , TGR5, which is present in the plasma membrane of brown adipose tissue and skeletal muscle tissue, is activated in brown adipose tissue and skeletal muscle tissue, type 2 (Tatsuo et al., 2001; Gatsios, P., Rupprecht, iodothyronine deiodinase (D2) (Brufau, G., Bahr, MJ, Staels, B., Claudel, T., et al., Nutr. Metab. 2010, 7, 73).

These effects comprehensively amplify glucose-dependent insulin secretion mediated by GLP-1 secretion from intestinal endocrine cells and promote insulin gene expression, enhancement of pancreatic beta cell proliferation, enhancement of pancreatic beta cell survival, Diabetes mellitus can be treated through inhibition of glucagon secretion. Hepatocyte can also treat liver disease by increasing the production of endothelial nitric oxide synthase (eNOS) and nitric oxide. Obesity can also be treated through activation of type 2 iodothyronine deiodinase (D2) in brown adipose tissue and skeletal muscle tissue.

GLP-1 secreted from the endocrine endocrine cells by the activation of TGR5 has been shown to induce glucose-dependent insulin secretion stimulation, insulin gene expression enhancement, pancreatic beta cell proliferation enhancement effect, pancreatic beta cell survival enhancement effect, glucagon secretion inhibitory effect, And it is known to be involved in the effect of treating obesity by slowing down the rate of emptying the stomach, suppressing the appetite, promoting the satiety, and suppressing the food intake.

Glucagon-like peptide-1 infusion improves left ventricular function through cardioprotective effect of cardiomyocytes from patients with ischemia and cardiopulmonary dysfunction. (2006) 12: 694-699., Ban, K., et. al . , Cardioprotective and vasodilatory actions of glucagon-like peptide-1 receptors are mediated through both glucagon-like peptide-1 receptor-dependent and -independent pathways. Circulation (2008) 117: 2340-2350. ), Pancreas and small intestine are known to be involved in efficient digestion and absorption regulation by promoting digestive enzyme secretion.

Glucagon-like peptide-1 (GLP-1) and its split products GLP-1 (9-37) and GLP-1 (28-37) stabilize atherosclerotic lesions in apoe ^{- / -} mice. Atherosclerosis (2013) 231, 427-435], the therapeutic effect of cerebrovascular diseases and neuronal inflammation (Salcedo I et al., Neuroprotective and neurotrophic actions of glucagon-like peptide-1 (GLP-1): an emerging opportunity to treat neurodegenerative and cerebrovascular disorders. British Journal of Pharmacology (2012) 166, 1586-1599).

The GLP-1 activates TGR5 and GPR119, one of the G protein-coupled receptors (GPCRs) (Reimann, F., et al . , Glucose sensing in L cells: a primary cell study. Cell Metab. (2008) 8: 532-539; Lauffer, LM, et al . , GPR119 is essential for oleoylethanolamide-induced glucagon-like peptide-1 secretion from the intestinal enteroendocrine L-cell. Diabetes (2009) 58: 1058-1066. ) Or activation of α-gustducin (Jang, HJ, et al . , 2007. Gut expressed gustducin and taste receptors regulate secretion of glucagon-like peptide-1. Proceeding of the National Academy of Science 104, 1506915074.). In particular, the activation of G protein-coupled receptor (GPCR) TGR5 (GPR131) expressed in brown adipose tissue and muscle increases energy expenditure and results in the treatment of obesity, (See Lieu T et al., GPBA: AG protein-coupled receptor for bile acids and an emerging therapeutic target for disorders of digestion and sensation. British Journal of Pharmacology (2013) in press).

In addition, GLP-1 is known to be capable of treating intestinal motility disorders, intestinal fluid and electrolyte secretion disorders (Lieu T et al., GPBA: AG protein-coupled receptor for bile acids and an emerging therapeutic target for disorders digestion and sensation, British Journal of Pharmacology DOI: 10.1111 / bph.12426).

There has been a continuing effort worldwide to develop drugs that are effective in serious diseases such as diabetes and cardiovascular disease. However, it is not easy to develop drugs with very low effects, such as obesity, hepatotoxicity, and edema, which are very effective.

On the other hand, medicines having anti-obesity effect are used for prevention or treatment of obesity, but these drugs are known to cause side effects such as oil stomach, abdominal bloating, dizziness, dryness, constipation, Development of strong natural functional materials is urgently required.

Accordingly, the technical problem to be solved by the present invention is to provide a pharmaceutical or food composition which is safe and has excellent side effects and is excellent in TGR5 agonist activity and can exhibit various useful effects.

In particular, another technical problem to be solved by the present invention is to provide an anti-diabetic effect; Anti-obesity effect; Neuronal protection effect; Or digestion and absorption control effects; Or a liver disease, or a pharmaceutical composition or food composition exhibiting at least one of the effects of improving or treating heart disease.

Another object of the present invention is to provide an anti-diabetic effect in an individual or patient comprising administering an effective amount of a pharmaceutical or food composition containing a safe extract as an active ingredient to a patient or patient in need of treatment or improvement; Anti-obesity effect; Neuronal protection effect; Or digestion and absorption control effects; Or liver disease, or a method for achieving at least one of the effects of improving or treating heart disease.

In order to accomplish the above object, the present invention provides a method for producing a plant of the genus Erigeron annuus , Sasa borealis (Hack.) Makino, Chamaecyparis obtusa fruit, Ricinus communis L., Solanum nigrum L., , Angelica dahurica B. et H. and Zanthoxylum piperitum AP DC) as an active ingredient, and an extract of a plant selected from the group consisting of Stellaria alsina var. Undulata (Thunb.) Ohwi, Angelica dahurica B. et H. and Zanthoxylum piperitum AP DC. And has TGR5 agonist activity, resulting in an antidiabetic effect; Anti-obesity effect; Neuronal protection effect; Or digestion and absorption control effects; Or a liver disease, or a composition for improving or treating a heart disease.

The present inventors have found that, among the natural products which have been used for a long time and proved to be safe, Erigeron annuus , Sasa borealis (Hack.) Makino, Chamaecyparis obtusa fruit, Ricinus communis L., Solanum nigrum L. Wherein at least one extract selected from the group consisting of Stellaria alsine var. Undulata (Thunb.) Ohwi, Angelica dahurica B. et H., and Zanthoxylum piperitum AP DC. The present invention has been completed.

&Quot; Treatment " in the present invention is an approach to obtaining beneficial or desired clinical results, which may or may not be able to detect beneficial or desired clinical results for the purposes of the present invention, whether or not partial But are not limited to, alleviation of the disease state, reduction in the degree of disease, stabilized (i.e., less severe) condition of the disease, delay or rate reduction of disease progression, improvement or temporary alleviation and alleviation of the disease state. In addition, treatment is an intervention performed with the intention of preventing the development or alteration of a lesion of one disease, which also means prolonged survival as compared to predicted survival when treatment is not accommodated. Thus, treatment refers to both therapeutic treatment and prophylactic measures, and those that need treatment include conditions that already have the disease, as well as conditions in which the disease is to be prevented.

As used herein, the term " improvement " is used in a broad sense to include alleviation and relief of symptoms after use of the composition of the present invention prior to use of the composition of the present invention.

Erigeron annuus ) is a plant of Asteraceae, which is known to be effective against diarrhea and fever symptoms caused by enteritis, and inflammation of the gums.

Sasa borealis (Hack.) Makino) is a major plant and is also known as "Sanjuk," and is known to have excellent antipyretic action. The present invention can preferably use a sage leaf.

Cloning ( Chamaecyparis obtusa ) is a plant of the spruce tree, and the bark or leaf part of the white wool is used for various purposes, and the leaf is known to have excellent hemostatic function. For the purpose of the present invention, it is preferred to use the fruit of the whitewood tree, in particular, of the whitewood tree.

Castor ( Ricinus communis L.) is a major pole and plant, also known as aquatic fungus, is known to have an excellent effect in removing inflammation.

Solanum nigrum L.) is a branch and plant that can be harvested in summer and autumn, and is known to be particularly effective in the womb health of middle-aged women.

Flea Herbs ( Stellaria There is alsine . undulata (Thunb.) Ohwi) is a plant of the japonica, also called an ant needle. It grows frequently in rice fields and fields. It is 15 ~ 25cm high and has no hairs. It grows as a big abandon because it spreads from branch to branch.

It is known as Angelica dahurica B. et H., which is also called as a white flower, and it is known that it has good effect on relieving pain and making blood circulate well. Particularly, for the purpose of the present invention, it is particularly preferable to use the roots of the peel.

Zanthoxylum piperitum AP DC. Has been known to have the effects of mungbean and vegetation, dehumidification, insect repellent, analgesia and so on. For the purpose of the present invention, it is particularly preferred to use the seed portion of the cherry fruit.

In one embodiment of the present invention, Erigeron annuus ), Sasa borealis (Hack.) Makino), Cliché ( Chamaecyparis obtusa ) fruit, castor ( Ricinus communis L., Solanum nigrum L.), flea herb ( Stellaria There is alsine . undulata (Thunb.) Ohwi), guritdae (Angelica B. H. et dahurica), and Chopi (Zanthoxylum piperitum AP DC.). The present inventors have found through experiments that the effect of any one or more of the extracts selected from the group consisting of the extracts of the present invention on insulin gene expression, glucagon secretion inhibition effect and blood glucose lowering effect on the prevention and improvement of diabetes It provides usages.

In another embodiment of the present invention, any one or more of the extracts selected from the group consisting of pomegranate, sage, cottonseed, castor, camphor, flea herb, gherkin and papaya seed has an appetite suppressing effect and slows down the rate at which the stomach is emptied And it is found through experiments that it promotes satiety, and provides an application for the anti-obesity effect through the suppression of the appetite of these extracts.

In another embodiment of the present invention, any one or more of extracts selected from the group consisting of amphibians, sorghum, wheat germ, castor, camphor, flea herb, gherkin, and camphor seed enhances digestive enzyme secretion in the pancreas and small intestine, Activity and mobility of the extracts through experimentation, thereby providing an application for improving the digestive and absorption disorders of these extracts.

According to another embodiment of the present invention, at least one extract selected from the group consisting of an amphipod, a sage, a cottonseed, a castor, a camphor, a flea herb, a peper, And the use of these extracts for improving heart disease is provided. More specifically, these extracts of the present invention can protect cardiac cells from ischemia and exhibit cardiac function-enhancing effects in patients suffering from cardiac arrest.

In another embodiment of the present invention, the brain's arcuate nucleus in hypothalamus (ARC) is activated by one or more extracts selected from the group consisting of amphipods, sage, cottonseed, castor, camphor, flea herb, , And vagus nerves, and provides a use for the neuroprotective effect of these extracts. More specifically, any one or more of the extracts selected from the group consisting of the herb of the present invention, sage, cottonseed, castor, camphor, flea herb, gherkin and papaya seed may be used for treating neurodegenerative diseases and cerebrovascular diseases , Neuroinflammation, and neurogenesis (Neurogenesis) can be expected to enhance the effect.

Another embodiment of the present invention provides the use of any one or more of extracts selected from the group consisting of amphipods, bamboo shoots, cotton buds, castor beans, flax seeds, flea herbs, More specifically, the plant extract of the present invention can be used for the treatment and / or prevention of hepatic inflammation, improvement of enterohepatic blood flow, improvement of liver function, treatment of cholestatic liver diseases, pruritus, Painless jaundice treatment effect, intestinal function improvement effect, neurotransmitter promotion effect can be expected.

Specifically, in one embodiment of the present invention, it was experimentally found that one or more extracts selected from the group consisting of Anopheles melliferae, Sorghum, Cotyledon fruit, Castor, Radish, Flea spp., Glycyrrhiza and Grasshopper stimulates GLP-1 secretion Lt; RTI ID = 0.0 > GLP-1 < / RTI > secretion effect of these extracts.

Any one or more plant extracts selected from the group consisting of herbaceous ginseng, sorghum, sorghum, cottonseed, castor, camphor, flea herb, gherkin and papaya seed of the present invention means extracts extracted by a conventional method in the field of the present invention, Followed by further fractionation, separation, filtration and the like.

For example, the plant extract according to the present invention may be any one or more of the solvent extracts or a dried product thereof selected from the group consisting of apricot, sage, cottonseed, castor, lobster, flea, When extracting the plant extract, the extraction solvent may be any solvent used for extracting natural products without limitation, for example, purified water; Lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; Hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether and dichloromethane; Or a mixed solvent thereof may be used, but it is preferable to use purified water, a lower alcohol having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol, butyl alcohol, or a mixed solvent thereof, and it is preferable to use purified water, ethanol, , And it is more preferable for the purpose of the present invention to use a mixed solvent of purified water and ethanol.

The amount of the extracting solvent may be 10 to 100 times, preferably 20 to 80 times, the weight of any one or more extracts selected from the group consisting of amaranths, sorghum, cottonseed, castor, lobster, flea, And more preferably 30 to 50 times, but is not limited thereto.

The method of extracting the plant extract can be performed by conventional extraction methods such as stirring, cold extraction, hot water extraction, immersion extraction, supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction, It is preferable to heat and reflux or extract at room temperature. The extraction temperature is preferably 10 to 30 ° C, and the extraction time is preferably 1 to 20 days, more preferably 1 to 5 days, but is not limited thereto.

More specifically, after the plants are shredded and ground, the extraction solvent is added in an amount of 1 to 20 times the volume of any one or more of the plant pulverized products selected from the group consisting of apricot, sage, cotton seed, castor, lentil, flea, (Decompression), followed by drying or purification. More specifically, an extraction solvent is added to extract an active ingredient substance as in the above-mentioned method, (a) heating at a temperature of 50 to 100 ° C for 1 to 20 hours in a state where a cooling condenser is installed to prevent evaporation of the solvent, (B) immersing the mixture at a temperature of 5 to 37 DEG C for 1 to 15 days to extract the active ingredient.

According to another embodiment of the present invention, the plant extract for the intestinal endocrine cell activation may further comprise one or more extracts selected from the group consisting of an amphipathic syrup, a sage, a flaxseed, a castor, a flaxseed, a flea sprout, Lt; / RTI >

The fraction of the present invention can be obtained by fractionating the plant extract (preferably a lower alcohol extract having 1-4 carbon atoms) with a solvent having a lower polarity than water such as hexane, chloroform, butanol and ethyl acetate, Chloroform may be used, but is not limited thereto.

In particular, for the purpose of achieving the effects of the present invention, the plant extract according to the present invention is preferably a hexane or chloroform fraction of a lower alcohol extract having 1-4 carbon atoms.

The fraction may be prepared by suspending or dissolving the plant extract in liquid form prior to drying or in a suitable solvent such as water, ethanol, or a mixed solvent thereof, and then dissolving the dried plant extract in a suitable solvent such as hexane, chloroform, butanol or ethyl acetate This low solvent may be sequentially added to separate each layer, and then each layer may be obtained by concentration under reduced pressure and / or drying.

At this time, the vacuum concentration is preferably performed using a vacuum rotary evaporator, but not limited thereto.

In addition, " drying " according to the present invention may be vacuum drying, vacuum drying, boiling drying, spray drying, room temperature drying or freeze-drying, However, the present invention is not limited to this drying method.

The present invention relates to a pharmaceutical composition comprising at least one extract selected from the group consisting of a herbicide, a sage, a cottonseed, a castor, a camphor, a flea herb, a pea, and a pea seed to promote secretion of the endocrine hormone and a neuropeptide, Anti-obesity effect; Neuronal protection effect; Or digestion and absorption control effects; Or a liver disease, or a pharmaceutical composition having an effect of improving or treating a heart disease or the like or the like.

The present invention also relates to an anti-diabetic effect comprising, as an active ingredient, one or more extracts selected from the group consisting of gabbroside, sorghum, cottonseed, castor, lobster, flea, Anti-obesity effect; Neuronal protection effect; Or digestion and absorption control effects; Or a liver disease, or a food composition having an effect of improving or treating heart disease or the like or the like.

That is, the composition according to the present invention can be used for medicines, health supplements, food additives, feed additives, and the like, but is not limited thereto.

The pharmaceutical compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of medicaments. The pharmaceutical composition according to the present invention can be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like oral preparation, external preparation, suppository and sterilized injection solution according to a conventional method Can be used. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, (sucrose), lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

More specifically, for example, powders can be prepared by simple mixing of an extract of the present invention with an appropriate pharmaceutically acceptable excipient such as lactose, starch, microcrystalline cellulose and the like. The granule is an extract of the present invention; Suitable excipients which are pharmaceutically acceptable; And a suitable pharmaceutically acceptable binder such as polyvinylpyrrolidone or hydroxypropylcellulose, followed by wet granulation using a solvent such as water, ethanol or isopropanol or dry granulation using a compressive force . Further, the tablet may be prepared by mixing the above granule with a suitable pharmaceutically acceptable salt such as magnesium stearate and then tableting it using a tablet machine.

The composition of the present invention may be administered orally or parenterally (for example, intramuscular injection, intraperitoneal injection, intravenous injection, infusion, subcutaneous injection, implant), inhalant, nasal administration agent, , Rectal, sublingual, transdermal, topical, and the like, but is not limited thereto. May be formulated into suitable dosage unit formulations, including those conventionally used and non-toxic, pharmaceutically acceptable carriers, additives, vehicles according to the route of administration. Depot formulations that are capable of sustained release of the drug over a period of time are also within the scope of the present invention.

Antidiabetic effect; Anti-obesity effect; Neuronal protection effect; Or digestion and absorption control effects; Or liver disease, or a cardiovascular disease, the plant extract according to the present invention may be administered at a daily dose of about 0.001 mg / kg to about 10 g / kg, A daily dosage of about 1 g / kg is preferred. However, the dosage may be adjusted depending on the degree of purification of one or more extracts selected from the group consisting of ganoderma, ganoderma, sorghum, castor, castor, flea, ganoderma, And the severity of the condition. For convenience, the total daily dose may be divided as needed and divided into several doses throughout the day.

When the extract of the present invention is used as a food composition, there is no particular limitation on the type of the food, when the extract of the present invention is used as a food composition, which is selected from the group consisting of pomegranate, sage, cottonseed, castor, Examples of the food which can be added with any one or more of extracts selected from the group consisting of the above-mentioned extracts selected from the group consisting of ginseng, sorghum, sorghum, cottonseed, cashew, flea sprouts, gherkin and chaffee seeds include drink, meat, sausage, bread, biscuit, rice cake, , Dairy products including candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, beverages, alcoholic beverages and vitamin complexes.

Any one or more extracts selected from the group consisting of herbaceous ginseng, sorghum, sorghum, cottonseed, castor, flea, flea, gherkin and chaffia seed of the present invention can be added directly to food or used together with other food or food ingredients, And the like. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose (for prevention or improvement). Generally, the amount of the extract in the food may be added in an amount of 0.01 to 50% by weight of the whole food. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

The health functional beverage composition of the food composition according to the present invention is characterized in that it contains at least one extract selected from the group consisting of the above-mentioned poppies, sage, sorghum, castor, camphor, flea sprout, There are no particular restrictions on the other components other than the above, and it may contain various flavors or natural carbohydrates as additional components such as ordinary beverages. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The flavoring agent may be a natural flavoring agent (tau martin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavorings (for example, saccharin and aspartame).

The extract of any one or more selected from the group consisting of the herb of the present invention, bamboo sage, cotton wool, castor, camphor, flea herb, gherkin, and chinese mackerel can be used as various nutrients, vitamins, minerals (electrolytes), synthetic flavors, Used in flavoring agents, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, Carbonating agents, and the like.

The present invention also relates to a method of treating or ameliorating a therapeutically effective amount or an amount effective for improving health of any one or more of the extracts selected from the group consisting of the above mentioned populations, such as Sorghum, Sorghum, Cotyledon fruit, Castor, Diabetic treatment or prevention, including administration to a patient or individual in need thereof; Obesity treatment or prevention; Nerve cell protection; Or regulation of digestion and absorption; Or liver disease, or a method of treating or ameliorating a heart disease.

That is, the present invention relates to a method of treating or preventing diabetes mellitus, comprising: extracting at least one selected from the group consisting of anoptera, a bamboo shoot, a cotton bud, a castor, a fagot, a flea bean, Obesity treatment or prevention; Nerve cell protection; Or regulation of digestion and absorption; Or liver disease, or heart disease improvement or treatment medicinal or health functional food use.

The present invention relates to a method for the treatment or prevention of diabetes mellitus of any one or more extracts selected from the group consisting of amphipods, sage, cottonseed, castor, lobster, flea sprouts, Obesity treatment or prevention; Nerve cell protection; Or regulation of digestion and absorption; Or liver disease, or heart disease improvement or treatment medicinal or health functional food use. The pharmaceutical composition or the food composition according to the present invention can be used as a medicinal composition for preventing and improving diabetes, suppressing appetite, anti-obesity, improving heart disease, improving digestion and absorption disorder, The treatment or improvement effect is excellent.

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

Experimental Method

1. Measurement of TGR5 activity

1) Cell culture

CHO-K1 cells were cultured in Ham's F-12K medium containing 10% (v / v) fetal bovine serum (FBS), 100 U / mL penicillin, and 100 μg / mL streptomycin, .

2) Measurement of TGR5 activity

The hTGR5 plasmid DNA was purchased from Origene, Inc. The transcript variant 3 was purchased from Origene Inc., and the vector was pCMV6-XL5 (4.5 kb), and the insert, GPBAR1 (SEQ ID NO: TGR5) was 1.4 Kb. In order to obtain the TGR5 plasmid, plasmids were obtained using Invitrogen Purelink (High pure filter maxiprep kit) after transfection into transformed E. coli containing 100 μg / ml ampicillin. 400ng / well of TGR5 expression plasmid (pCMV6-XL5 / TGR5) was transfected with CHO-K1 using Neon ^™ (Invitrogen, Cergy Pontoise, France) and treated for 72 hrs. cAMP was measured using a cAMP dynamin kit (Cisbio International).

2. GLP-1 secretion stimulation experiment using NCI-H716 cells

1) Cell culture

Human-derived cell lines, NCI-H716 cells, were obtained from the American Type Culture Collection (Manassas, Va.). Cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 IU / ml penicillin, 100 / ml streptomycin.

2) Determination of GLP-1 secretion effect of sample

NCI-H716 cells were cultured in 96-wells at a concentration of 2 x 10 < ⁵ > cells / plate for 3 days. On the day of the experiment, the cells were rinsed with phosphate-buffered saline (PBS), washed with Krebs-Ringer bicarbonate buffer (KRB, 128.8 mmol / l NaCl, 4.8 mmol / l KCl, 1.2 mmol / l KH ₂ PO ₄ , 1.2 mmol / 1 mM MgSO ₄ , 2.5 mmol / l CaCl ₂ , 5 mmol / l NaHCO ₃ , and 10 mmol / l HEPES, 0.2% BSA, pH 7.4) and incubated at 37 ° C for 1 hour. After 1 hour incubation, the supernatant was collected (100 μL) for GLP-1 analysis and analyzed by GLP-1 activity ELISA kit (EGLP-35K, Millipore). The concentration of GLP-1 was calculated according to the supplied GLP-1 standard (pmole / mL), and the result was compared with the concentration secreted by stimulation of GLP-1 secreted by the stimulus of the sample, of control.

3) Measurement of changes in intracellular calcium concentration

Fluorescence-based assay was used to examine changes in intracellular calcium. Cells were collected at a concentration of 4.0 × 10 ⁴ cells / well in a 96-well plate. Fluorometric imaging plate reader (FLIPR ^™ ) was prepared using a calcium-5 assay kit purchased from Molecular Devices, Intracellular Ca ²⁺ concentration was measured using Flexstation III (Molecular Devices Co., Sunnyvale, Calif., USA). Fluorescence intensity of calcium stained for 120 seconds after injection of sample was measured at 488 nm excitation and 525 emmision. The results were as follows: maximum fluorescence value - minimum fluorescence value = relative fluorescence value And was automatically calculated as a fluorescence unit (delta relative fluorescent units (? RFU)).

Activation of the GPCRs activates the G protein complex (Gα-gustducin, Gα-transtducin, Gα14, Gβ3, Gβ1, Gγ13), which activates the phospholipase to activate the type Ⅲ IP3-receptor, it is possible to measure the activation of GPCRs by measuring the change of intracellular Ca ²⁺ ion concentration because it causes increase of Ca ²⁺ concentration in cytoplasm.

In addition, when TRP channels are activated, the calcium ions outside the cell membrane enter the cells, so that the measurement of the intracellular Ca ^{2 +} ion concentration may be able to measure the activation of TRP channels.

4) cAMP measurement method

NCI-H716 cell line in 24-well 1 x 10 ⁶ cells / mL concentration of culture and then cultured for 3 days. Experiment were cultured in the same day a sample and with 5% CO ₂ incubator in ℃ 37 in KRB buffer, the reaction medium is removed and cells are subjected to back washing with water three times to process the KRB buffer into 0.1N HCl 0.5mL 150 rpm, at room temperature The reaction was carried out for 20 minutes. The reacted cells were centrifuged at 1000 g for 10 minutes, and the supernatant was obtained. The cyclic AMP kit using enzyme immunoassay (EIA) was used.

5) Results

The concentrations of GLP-1 and cAMP were determined according to the EIA method. Changes in intracellular calcium concentration were expressed in relative fluorescence units (ΔRFU). All values were expressed as mean ± SEM (n = 9) in triplicate experiments. * p &lt;0.05; ** p &lt;0.01; *** p <0.001 vs control (control group)

&Lt; Example 1 >

1. Preparation of Angelica gigantosa Extract

Except for roots, the outposts were purchased from the Sichuan Won (370-114, North Hwasun, Hwasun Gun, Jeonnam), lyophilized, ground with a grinder, and 40 mL of 50 wt% ethanol was added per 1 g of the sample. The extract was filtered twice using a filter paper (Advantec No. 2, 4, Japan), concentrated using a rotary evaporator (B-480, Buchi, Switzerland), lyophilized again, Respectively.

2. Measurement of TGR5 activity

2-1. Effect of 50% Ethanol Extract of Angelica gigas on Growth Activity of TGR5 in Cells Transfected with hTGR5 in CHO-K1 Cells

To the culture medium of CHO-K1 cells transfected with the TGR5 plasmid, 50% ethanol extract of Angelica gigas was added for 1 hour. TGR5 activity was assessed by Luciferase assay as mentioned in the previous method and all values are expressed as means ± SEM. n = 9. * p <0.05, ** p <0.01, *** p <0.001 vs control (control group)

sample density (fold of control) Negative control group 1.000 ± 0.144 Extract (50% ethanol extract) 10 μg / mL 1.407 + 0.096 50 μg / mL 1.717 + - 0.399 ^* 100 μg / mL 3.071 0.358 ^*** Lithocholic acid (LCA) 0.1 μM 1.536 + 0.180 1.0 [mu] M 2.777 ± 0.303 ^*** 5.0 μM 4.506 ± 0.413 ^***

As shown in Table 1, the TGR5 activity was increased by 50% ethanol extract of Angelica keiskei on the cells transfected with hTGR5. From these results, it can be concluded that the herb extract of the present invention may exhibit an anti-obesity effect, a liver disease treatment effect, a heart disease treatment effect, or a gastrointestinal disease treatment effect.

3. Measurement of GLP-1 secretion enhancing activity

3-1. NCI - H716  Extract of 50% ethanol extract from the cell GLP -1 secretion effect

sample density GLP-1 (fold of control) Control group 1.000 + 0.088 Extract (50% ethanol extract) 500 μg / mL 2.320 0.481 ^* Lithocholic acid (LCA) 100 μg / mL 1.282 + 0.087 ^* 250 μg / mL 2.172 + 0.187 ^** 500 μg / mL 4.025 ± 0.413 ^***

As shown in the above Table 2, it was found that GLP-1 secretion was effective at 500 μg / mL concentration in NCI-H716 cells. Thus, the 50% ethanol extract of Ganoderma lucidum can activate the TGR5 expressed in the intestine and secrete GLP-1, thereby slowing down the gastric emptying, and promoting obesity by controlling the appetite suppression and satiety. Neuroprotective and neurotrophic actions such as treatment of neurodegenerative and cerebrovascular disorders, neuroinflammation therapy, and neurogenesis enhancing effects can be expected.

3-2. NCI - H716  Extract of 50% ethanol extract from the cell Intracellular Ca ^{2 +}  Concentration change

sample density In Ca ^{2 +} concentration in the cell (ΔRFU) Control group 9.181 ± 0.632 Extract (50% ethanol extract) 500 μg / mL 30.927 + 1.050 ^** Lithocholic acid (LCA) 5 [mu] M 10.647 ± 0.366 ^** 10 μM 19.895 ± 0.509 ^*** 30 μM 56.615 ± 2.626 ^***

According to the results shown in Table 3, 500 μg / mL of 50% ethanol extract of Angelica keiskei koidzii significantly increased intracellular calcium concentration compared with the control group. The 50% ethanol extract of Angelica gigas seemed to increase GLP-1 secretion by increasing intracellular calcium concentration.

3-3. NCI - H716  Extract of 50% ethanol extract from the cell Intracellular cAMP  Increase effect

sample density Changes in intracellular cAMP concentration Control group 1.000 ± 0.101 Extract (50% ethanol extract) 500 μg / mL 3.067 ± 0.601 ^* Lithocholic acid (LCA) 5 [mu] M 3.962 + 0.084 ^*** 10 μM 7.665 ± 1.375 ^** 30 μM 13.823 ± 1.260 ^***

As shown in Table 4 above, it was shown that apricot (50% ethanol extract) was more effective in increasing intracellular cAMP than the control group. In other words, cAMP concentration, which is one of signal transduction molecules of GPCRs, is measured and it is thought that the extract of Anopheles sinensis has an effect on the increase of GLP-1 secretion.

&Lt; Example 2 > Extract of Saururus chinensis

1. Manufacture of leaf extract

The leaves of the ganoderma lucidum (the outposts except for the roots) were purchased from Samcheonwon (370-114, Noppyeong-myeon, Hwasun-gun, Jeonnam), lyophilized and pulverized by a pulverizer. 40 ml of 50 wt% ethanol was added per 1 g of the sample and extracted at room temperature for 3 days. The extract was filtered twice using a filter paper (Advantec No. 2, 4, Japna), concentrated using a rotary evaporator (B-480, Buchi, Switzerland), lyophilized again, Respectively.

2. Measurement of TGR5 activity

2-1. Enhancement of TGR5 activity of 50% ethanol extract of Sanjiku leaves on cells transfected with hTGR5 in CHO-K1 cells

To the culture of CHO-K1 cells transfected with the TGR5 plasmid, 50% ethanol extract of Sanjiku leaf was added for 1 hour. TGR5 activity was assessed by Luciferase assay as mentioned in the previous method and all values are expressed as means ± SEM. n = 9. * p <0.05, ** p <0.01, *** p <0.001 vs control (control group)

sample density (fold of control) Control group 1.000 ± 0.144 Sunflower leaf (50% ethanol extract) 100 μg / mL 1.877 0.358 ^*** Lithocholic acid (LCA) 0.1 μM 1.536 + 0.130 1.0 [mu] M 2.777 ± 0.303 ^*** 5.0 μM 4.506 ± 0.413 ^***

As shown in Table 5, TGR5 activity was increased by the 50% ethanol extract of Sanjiku leaf in hTGR5 transfected cells.

3. Measurement of GLP-1 secretion enhancing activity

3-1. NCI - H716  In a cell Ganjuk Leaf  Of 50% ethanol extract GLP -1 secretion effect

sample density GLP-1 (fold of control) Control group 1.000 + 0.038 Sunflower leaf (50% ethanol extract) 250 μg / mL 2.761 + - 0.314 ^** Lithocholic acid (LCA) 100 μg / mL 1.282 + 0.087 ^* 250 μg / mL 2.172 + 0.187 ^** 500 μg / mL 4.025 ± 0.413 ^***

As shown in Table 6 above, the leaf of Sanjiku (50% ethanol extract) showed GLP-1 secretion effect at 250 μg / mL concentration in NCI-H716 cells.

3-2. NCI - H716  In a cell Ganjuk Leaf  Of 50% ethanol extract Intracellular Ca ^{2 +}  Concentration change

sample density In Ca ^{2 +} concentration in the cell (ΔRFU) Control group 9.181 ± 0.632 Sunflower leaf (50% ethanol extract) 250 μg / mL 34.149 ± 2.740 ^*** Lithocholic acid (LCA) 5 [mu] M 10.647 ± 0.366 ^** 10 μM 19.895 ± 0.509 ^*** 30 μM 56.615 ± 2.626 ^***

According to the results shown in Table 7, 250 μg / mL of the 50% ethanol extract of Ganoderma lucidum significantly increased the intracellular calcium concentration in the cells compared to the control. The 50% ethanol extract of Ganjuk leaves was thought to increase GLP-1 secretion by increasing intracellular calcium concentration.

&Lt; Example 3 >

1. Preparation of Cotton White Fruit Extract

The fruit of Prunus yedoensis was purchased from Sanchuanwon (370-114 Songun Road, Hwasun Gun, Jeonnam), lyophilized and pulverized by a pulverizer, and 40 mL of 50% by weight ethanol was added per 1 g of sample and extracted at room temperature for 3 days. The extract was filtered twice using a filter paper (Advantec No. 2, 4, Japna), concentrated using a rotary evaporator (B-480, Buchi, Switzerland), lyophilized again, Respectively.

2. Measurement of TGR5 activity

2-1. Enhancement of TGR5 activity of 50% ethanol extract of white flour on cells transfected with hTGR5 in CHO-K1 cells

To the culture of CHO-K1 cells transfected with the TGR5 plasmid, a 50% ethanolic extract of white cotton fruit was added for 1 hour. TGR5 activity was assessed by Luciferase assay as mentioned in the previous method and all values are expressed as means ± SEM. n = 9. * p <0.05, * p <0.01, *** p <0.001 vs control (control group)

sample density (fold of control) Control group 1.000 ± 0.144 Cottonseed fruit (50% ethanol extract) 100 μg / mL 1.502 + - 0.148 ^* Lithocholic acid (LCA) 0.1 μM 1.536 + 0.130 1.0 [mu] M 2.777 ± 0.303 ^*** 5.0 μM 4.506 ± 0.413 ^***

As shown in Table 8, the activity of TGR5 was increased by 50% ethanol extract of the white cotton fruit in the cells transfected with hTGR5.

3. Measurement of GLP-1 secretion enhancing activity

3-1. NCI - H716  In a cell Pruning fruit  Of 50% ethanol extract GLP -1 secretion effect

sample density GLP-1 (fold of control) Control group 1.000 + 0.038 Cottonseed fruit (50% ethanol extract) 250 μg / mL 2.725 +/- 0.039 ^*** Lithocholic acid (LCA) 100 μg / mL 1.282 + 0.087 ^* 250 μg / mL 2.172 + 0.187 ^** 500 μg / mL 4.025 ± 0.413 ^***

As shown in Table 9, it was found that GLP-1 secretion was effective at a concentration of 250 μg / mL in NCI-H716 cells. Thus, the 50% ethanol extract of the white fruit extract activates TGR5 expressed in the intestine and secretes GLP-1, thereby enhancing glucose-dependent insulin secretion stimulation in pancreas, promoting insulin gene expression, promoting pancreatic beta cell proliferation, Survival enhancement effect, glucagon secretion inhibitory effect, and the like can be expected to be effective for the treatment of diabetes.

3-2. NCI - H716  In a cell Pruning fruit  Of 50% ethanol extract Intracellular Ca ^{2 +}  Concentration change

sample density In Ca ^{2 +} concentration in the cell (ΔRFU) Control group 9.181 ± 0.632 Cottonseed fruit (50% ethanol extract) 250 μg / mL 59.252 ± 2.284 ^*** Lithocholic acid (LCA) 5 [mu] M 10.647 ± 0.366 ^** 10 μM 19.895 ± 0.509 ^*** 30 μM 56.615 ± 2.626 ^***

According to the results shown in Table 10, 250 μg / mL of the 50% ethanol extract of the whitespace fruit significantly increased the intracellular calcium concentration compared with the control group. The 50% ethanol extract of white flour was thought to increase GLP-1 secretion by increasing intracellular calcium concentration.

<Example 4> Castor (Castor) Leaf Extract

1. Preparation of Pharmacia Extract

The leaves of the castor were purchased from Samcheonwon (370-114 Songdang road, Hwasun Gun, Jeonnam), lyophilized, pulverized by a pulverizer, 40 ml of 50% by weight ethanol was added per 1 g of the sample and extracted at room temperature for 3 days. The extract was filtered twice using a filter paper (Advantec No. 2, 4, Japan), concentrated using a rotary evaporator (B-480, Buchi, Switzerland), lyophilized again, Respectively.

2. Measurement of TGR5 activity

2-1. Enhancement of TGR5 activity of 50% ethanol extract of castor leaves on cells transfected with hTGR5 in CHO-K1 cells

A 50% ethanol extract of castor leaves was added to the culture of CHO-K1 cells transfected with the TGR5 plasmid for 1 hour. TGR5 activity was assessed by Luciferase assay as mentioned in the previous method and all values are expressed as means ± SEM. n = 9. * p <0.05, * p <0.01, *** p <0.001 vs control (control group)

sample density (fold of control) Control group 1.000 ± 0.144 Castor (50% ethanol extract) 100 μg / mL 3.714 ± 0.827 ^** Lithocholic acid (LCA) 0.1 μM 1.536 + 0.130 1.0 [mu] M 2.777 ± 0.303 ^*** 5.0 μM 4.506 ± 0.413 ^***

As shown in Table 11, TGR5 activity was increased by 50% ethanol extract of castor in hTGR5-transfected cells.

3. Measurement of GLP-1 secretion enhancing activity

3-1. NCI - H716  50% Ethanol Extract of Castor from Cells GLP -1 secretion effect

sample density GLP-1 (fold of control) Control group 1.000 + 0.038 Castor (50% ethanol extract) 250 μg / mL 2.820 ± 0.133 ^*** Lithocholic acid (LCA) 100 μg / mL 1.282 + 0.087 ^* 250 μg / mL 2.172 + 0.187 ^** 500 μg / mL 4.025 ± 0.413 ^***

As shown in Table 12 above, castor (50% ethanol extract) showed GLP-1 secretion effect at 250 μg / mL concentration in NCI-H716 cells.

3-2. NCI - H716  50% Ethanol Extract of Castor from Cells Intracellular Ca ^{2 +}  Concentration change

sample density In Ca ^{2 +} concentration in the cell (ΔRFU) Control group 9.181 ± 0.632 Castor (50% ethanol extract) 250 μg / mL 41.731 ± 2.162 ^** Lithocholic acid (LCA) 5 [mu] M 10.647 ± 0.366 ^** 10 μM 19.895 ± 0.509 ^*** 30 μM 56.615 ± 2.626 ^***

According to the results shown in Table 13, 250 μg / mL of 50% ethanol extract of castor leaves significantly increased intracellular calcium concentration compared with the control group. The 50% ethanol extract of castor seemed to increase GLP-1 secretion by increasing intracellular calcium concentration.

&Lt; Example 5 >

1. Preparation of Extracts

The leaf and the fruit of the lobster were purchased from the prawn house (370-114, Hwasun-myeon, Hwasun-gun, Jeonnam), lyophilized and pulverized by a pulverizer. 40 mL of 50 wt% ethanol was added per 1 g of the sample and extracted at room temperature for 3 days. The extract was filtered twice using a filter paper (Advantec No. 2, 4, Japna), concentrated using a rotary evaporator (B-480, Buchi, Switzerland), lyophilized again, Respectively.

2. Measurement of TGR5 activity

2-1. Enhancement of TGR5 activity of 50% ethanol extract of black currants in cells transfected with hTGR5 in CHO-K1 cells

To the culture medium of CHO-K1 cells transfected with the TGR5 plasmid, 50% ethanol extracts of lambs were added for 1 hour. TGR5 activity was assessed by Luciferase assay as mentioned in the previous method and all values are expressed as means ± SEM. n = 9. * p <0.05, * p <0.01, *** p <0.001 vs control (control group)

sample density (fold of control) Control group 1.000 ± 0.144 Extract (50% ethanol extract) 100 μg / mL 4.274 + 1.045 ^* Lithocholic acid (LCA) 0.1 μM 1.536 + 0.130 1.0 [mu] M 2.777 ± 0.303 ^*** 5.0 μM 4.506 ± 0.413 ^***

As shown in Table 14 above, the TGR5 activity was increased by the 50% ethanol extract of Lambs on the cells transfected with hTGR5.

3. Measurement of GLP-1 secretion enhancing activity

3-1. NCI - H716  50% Ethanol Extract in Cells from Cells GLP -1 secretion effect

sample density GLP-1 (fold of control) Control group 1.000 + 0.038 Extract (50% ethanol extract) 100 μg / mL 2.469 ± 0.173 250 μg / mL 4.172 + 0.085 500 μg / mL 5.860 + 0.132 Lithocholic acid (LCA) 100 μg / mL 1.282 + 0.087 ^* 250 μg / mL 2.172 + 0.187 ^** 500 μg / mL 4.025 ± 0.413 ^***

As shown in Table 15 above, it was found that GLP-1 secretion effect was obtained at a concentration of 250 μg / mL in NCI-H716 cells from the white fruit (50% ethanol extract).

3-2. NCI - H716  50% Ethanol Extract in Cells from Cells Intracellular Ca ^{2 +}  Concentration change

sample density In Ca ^{2 +} concentration in the cell (ΔRFU) Control group 9.181 ± 0.632 Extract (50% ethanol extract) 100 μg / mL 19.520 ± 2.249 ^*** 250 μg / mL 60.531 + - 6.728 ^*** 500 μg / mL 120.347 +/- 3.425 ^*** Lithocholic acid (LCA) 5 [mu] M 10.647 ± 0.366 ^** 10 μM 19.895 ± 0.509 ^*** 30 μM 56.615 ± 2.626 ^***

According to the results shown in Table 16, 250 μg / mL of 50% ethanol extract of L. japonica significantly increased intracellular calcium concentration compared to the control group. The 50% ethanol extract of L. rabbit was thought to increase GLP-1 secretion by increasing intracellular calcium concentration.

3-3. NCI - H716  50% Ethanol Extract in Cells from Cells Intracellular cAMP  Increase effect

sample density Changes in intracellular cAMP concentration Control group 1.000 ± 0.101 Extract (50% ethanol extract) 100 μg / mL 1.920 + 0.084 ^** 250 μg / mL 2.620 + - 0.229 ^*** 500 μg / mL 6.470 ± 0.622 ^*** Lithocholic acid (LCA) 5 [mu] M 3.962 + 0.084 ^*** 10 μM 7.665 ± 1.375 ^** 30 μM 13.823 ± 1.260 ^***

As shown in Table 17 above, the 50% ethanol extract of Panax ginseng was more effective in increasing intracellular cAMP than the control group. In other words, cAMP concentration, which is one of the signal transduction molecules of GPCRs, was measured, suggesting that L. chamai extract has an effect on GLP-1 secretion.

Example 6: Flea extract

1. Preparation of flea extract

The flea herbs were purchased from Samcheonwon (370-114, Hwasun-myeon, Hwasun-gun, Jeonnam, Korea), lyophilized, and pulverized with a grinder. 40 ml of 50% by weight ethanol was added per 1 g of the sample and extracted at room temperature for 3 days. The extract was filtered twice using a filter paper (Advantec No. 2, 4, Japna), concentrated using a rotary evaporator (B-480, Buchi, Switzerland), lyophilized again, Respectively.

2. Measurement of TGR5 activity

2-1. Effect of 50% Ethanol Extract of Flea Grass on the TGR5 Activity in Cells Transfected with hTGR5 in CHO-K1 Cells

To the culture of CHO-K1 cells transfected with the TGR5 plasmid, 50% ethanol extract of flea herb was added for 1 hour. TGR5 activity was assessed by Luciferase assay as mentioned in the previous method and all values are expressed as means ± SEM. n = 9. * p <0.05, * p <0.01, *** p <0.001 vs control (control group)

sample density (fold of control) Control group 1.000 ± 0.144 Flea herb (50% ethanol extract) 100 μg / mL 2.618 ± 0.311 ^*** Lithocholic acid (LCA) 0.1 μM 1.536 + 0.130 1.0 [mu] M 2.777 ± 0.303 ^*** 5.0 μM 4.506 ± 0.413 ^***

As shown in Table 18, TGR5 activity was increased by 50% ethanol extract of flea seeds in cells transfected with hTGR5.

3. Measurement of GLP-1 secretion enhancing activity

3-1. GLP-1 secretion effect of 50% ethanol extract of flea spp. In NCI-H716 cells

sample density GLP-1 (fold of control) Control group 1.000 + 0.038 Flea herb (50% ethanol extract) 100 μg / mL 1.548 + 0.081 ^*** 250 μg / mL 2.014 ± 0.219 ^*** 500 μg / mL 5.934 + 0.094 ^*** Lithocholic acid (LCA) 100 μg / mL 1.282 + 0.087 ^* 250 μg / mL 2.172 + 0.187 ^** 500 μg / mL 4.025 ± 0.413 ^***

As shown in Table 19, the flea extract (50% ethanol extract) showed GLP-1 secretion at 100, 250, and 500 μg / mL concentration in NCI-H716 cells.

3-2. Intracellular Ca ^{2 +} concentration change of 50% ethanol extract from Herb flea NCI-H716 cells

sample density In Ca ^{2 +} concentration in the cell (ΔRFU) Control group 9.181 ± 0.632 Flea herb (50% ethanol extract) 100 μg / mL 18.885 ± 2.298 ^** 250 μg / mL 24.795 ± 1.033 ^*** 500 μg / mL 38.703 占 .445 ^*** Lithocholic acid (LCA) 5 [mu] M 10.647 ± 0.366 ^** 10 μM 19.895 ± 0.509 ^*** 30 μM 56.615 ± 2.626 ^***

According to the results shown in Table 20, 100%, 250, and 500 μg / mL of 50% ethanol extract of flea extracts significantly increased intracellular calcium concentration compared with the control group. The 50% ethanol extract of the flea herb was thought to increase GLP-1 secretion by increasing intracellular calcium concentration.

<Example 7> Bleaching (pale yellow) extract

1. Preparation of white paprika extract

The outpasted leaves were purchased from Samcheonwon (370-114 Songdang-road, Hwasun-gun, Jeonnam), lyophilized, ground with a pulverizer, and 40 ml of 50% by weight ethanol was added per 1 g of the sample. The extract was filtered twice using a filter paper (Advantec No. 2, 4, Japna), concentrated with a rotary evaporator (B-480, Buchi, Switzerland), lyophilized again and pulverized. Respectively.

2. Measurement of TGR5 activity

2-1. Enhancement of TGR5 activity of 50% ethanol extract of white papillary (Gurley) in cells transfected with hTGR5 in CHO-K1 cells

To the culture medium of CHO-K1 cells transfected with the TGR5 plasmid, 50% ethanol extract of white paper was added for 1 hour. TGR5 activity was assessed by Luciferase assay as mentioned in the previous method and all values are expressed as means ± SEM. n = 9. * p <0.05, * p <0.01, *** p <0.001 vs control (control group)

sample density (fold of control) Control group 1.000 ± 0.144 Blank paper (50% ethanol extract) 100 μg / mL 4.055 0.4851 ^*** Lithocholic acid (LCA) 0.1 μM 1.536 + 0.130 1.0 [mu] M 2.777 ± 0.303 ^*** 5.0 μM 4.506 ± 0.413 ^***

As shown in Table 21, TGR5 activity was increased by the 50% ethanol extract of white ginseng in cells transfected with hTGR5.

3. Measurement of GLP-1 secretion enhancing activity

3-1. GLP-1 secretion effect of 50% ethanol extract of white papilla (N. gonorrhoeae) on NCI-H716 cells

sample density GLP-1 (fold of control) Control group 1.000 + 0.038 Blank paper (50% ethanol extract) 100 μg / mL 1.258 + 0.072 ^*** 250 μg / mL 1.811 + 0.036 ^*** 500 μg / mL 2.023 + 0.032 ^*** Lithocholic acid (LCA) 100 μg / mL 1.282 + 0.087 ^* 250 μg / mL 2.172 + 0.187 ^** 500 μg / mL 4.025 ± 0.413 ^***

As shown in Table 22 above, it was found that GLP-1 secretion effect was obtained at 100, 250, and 500 μg / mL concentration of NCI-H716 cells on a white background (50% ethanol extract).

3-2. NCI - H716  In cells, Vet ) Of 50% ethanol extract Intracellular Ca ^{2 +}  Concentration change

sample density In Ca ^{2 +} concentration in the cell (ΔRFU) Control group 9.181 ± 0.632 Blank paper (50% ethanol extract) 100 μg / mL 15.319 + - 0.984 ^* 250 μg / mL 10.814 + - 0.852 500 μg / mL 26.727 ± 1.625 ^*** Lithocholic acid (LCA) 5 [mu] M 10.647 ± 0.366 ^** 10 μM 19.895 ± 0.509 ^*** 30 μM 56.615 ± 2.626 ^***

According to the results shown in Table 23, 500 μg / mL of 50% ethanol extract of white ginseng cells significantly increased intracellular calcium concentration compared with the control group. The 50% ethanol extract of the flea herb was thought to increase GLP-1 secretion by increasing intracellular calcium concentration.

&Lt; Example 8 >

1. Preparation of chopped P. extract

The chopped pine seeds were purchased from Samcheonwon (370-114, Noppyeong-myeon, Hwasun-gun, Jeonnam), lyophilized and ground with a pulverizer. 40 ml of 50% by weight ethanol was added per 1 g of the sample and extracted at room temperature for 3 days. The extract was filtered twice using a filter paper (Advantec No. 2, 4, Japan), concentrated using a rotary evaporator (B-480, Buchi, Switzerland), lyophilized again, Respectively.

2. Measurement of TGR5 activity

2-1. Enhancement of TGR5 activity of 50% ethanol extract of Chopo PC on CHO-K1 cells transfected with hTGR5

To the culture medium of CHO-K1 cells transfected with the TGR5 plasmid, 50% ethanol extract of Choc Pc was added for 1 hour. TGR5 activity was assessed by Luciferase assay as mentioned in the previous method and all values are expressed as means ± SEM. n = 9. * p <0.05, * p <0.01, *** p <0.001 vs control (control group)

sample density (fold of control) Control group 1.000 ± 0.144 Chopped (50% ethanol extract) 100 μg / mL 1.945 ± 0.230 ^** Lithocholic acid (LCA) 0.1 μM 1.536 + 0.130 1.0 [mu] M 2.777 ± 0.303 ^*** 5.0 μM 4.506 ± 0.413 ^***

As shown in Table 24, the activity of TGR5 was increased by the 50% ethanol extract of E.coli in hTGR5 transfected cells.

3. Measurement of GLP-1 secretion enhancing activity

3-1. GLP-1 secretion effect of 50% ethanol extract of Choc Pci in NCI-H716 cells

sample density GLP-1 (fold of control) Control group 1.000 + 0.038 Chopped (50% ethanol extract) 100 μg / mL 1.168 ± 0.177 ^** 250 μg / mL 1.981 ± 0.102 ^*** 500 μg / mL 2.697 + 0.069 ^*** Lithocholic acid (LCA) 100 μg / mL 1.282 + 0.087 ^* 250 μg / mL 2.172 + 0.187 ^** 500 μg / mL 4.025 ± 0.413 ^***

As shown in Table 25, it was found that GLP-1 was secreted at 100, 250, and 500 μg / mL concentration in NCI-H716 cells (50% ethanol extract).

3-2. NCI - H716  In a cell Chopec  Of 50% ethanol extract Intracellular Ca ^{2 +}  Concentration change

sample density In Ca ^{2 +} concentration in the cell (ΔRFU) Control group 9.181 ± 0.632 Chopped (50% ethanol extract) 100 μg / mL 13.746 + 0.681 250 μg / mL 16.362 ± 0.539 ^** 500 μg / mL 28.990 占 2.188 ^*** Lithocholic acid (LCA) 5 [mu] M 10.647 ± 0.366 ^** 10 μM 19.895 ± 0.509 ^*** 30 μM 56.615 ± 2.626 ^***

According to the results shown in Table 26, it was found that the concentration of 250 μg / mL of 50% ethanol extract of chopped PC significantly increased intracellular calcium concentration compared with the control. The 50% ethanol extract of Chuppia was thought to increase GLP-1 secretion by increasing intracellular calcium concentration.

Claims (12)

For anti-obesity containing the extract of Angelica gigas; as an active ingredient; Or for controlling digestion and absorption; Or liver disease, or a pharmaceutical or food composition for the treatment or prevention of heart disease. For protecting nerve cells containing Sasa extract as an active ingredient; Or for controlling digestion and absorption; Or liver disease, or a pharmaceutical or food composition for the treatment or prevention of heart disease. 3. The pharmaceutical or food composition according to claim 2, wherein the sage is a sage leaf. For anti-diabetic drugs containing an extract of Nasopharynx as an active ingredient; Or for controlling digestion and absorption; Or liver disease, or a pharmaceutical or food composition for the treatment or prevention of heart disease. For anti-obesity containing castor oil extract as an active ingredient; Or for controlling digestion and absorption; Or liver disease, or a pharmaceutical or food composition for the treatment or prevention of heart disease. The pharmaceutical or food composition according to claim 5, wherein the castor is a castor's blade For anti-obesity, which contains extracts of camphor; as an active ingredient; Or for controlling digestion and absorption; Or liver disease, or a pharmaceutical or food composition for the treatment or prevention of heart disease. For antidiabetic use containing flea extracts as an active ingredient; For anti-obesity; For nerve cell protection; Or for controlling digestion and absorption; Or liver disease, or a pharmaceutical or food composition for the treatment or prevention of heart disease. For anti-diabetic drugs containing grit extract as an active ingredient; For anti-obesity; For nerve cell protection; Or for controlling digestion and absorption; Or liver disease, or a pharmaceutical or food composition for the treatment or prevention of heart disease. For antidiabetic use containing extracts of mulberry leaves as an active ingredient; For anti-obesity; Or for controlling digestion and absorption; Or liver disease, or a pharmaceutical or food composition for the treatment or prevention of heart disease. 11. The pharmaceutical or food composition according to claim 10, wherein the plant is a plant. 12. A pharmaceutical or food product according to any one of claims 1 to 11, characterized in that the pomegranate, sorghum, sorghum, castor, hammerhead, flea herb, gherkin or mulberry extract have TGR 5 agonist activity Composition.