JP6273440B2 - GLP-1 production promoter, DPPIV inhibitor and glucose absorption inhibitor - Google Patents

Description

  The present invention relates to a GLP-1 production promoter, a DPPIV inhibitor, a glucose absorption inhibitor, and uses thereof, which comprise a specific plant or a processed product thereof as an active ingredient.

  Typical lifestyle-related diseases are heart disease, diabetes, dyslipidemia (hyperlipidemia), hypertension, stroke, etc. Many of these causes are related to obesity, especially by visceral fat accumulation I know it is being caused. The accumulation of visceral fat is due to the fact that sugar (glucose) in the blood is not consumed as active energy, and excess glucose is stored in the tissue as fat.

  In the mechanism of glucose metabolism in the living body, it has a function of regulating blood sugar concentration. When food is ingested, it is converted into fatty acids, amino acids, and glucose (glucose) through digestion processes such as carbohydrate, protein, and fat degrading enzymes. It is absorbed in the tube (intestinal tract). Glucose absorbed in the intestinal tract is released into the blood and temporarily increases the glucose concentration in the blood, so that the cells of the pancreas detect an increase in blood glucose level and release insulin, a secreted hormone, into the blood However, insulin works to take up glucose in the blood into liver cells, fat cells, and muscle tissue, and when glucose is stored in cells instead of glycogen (a high molecular weight product of glucose), the glucose concentration in the blood (blood glucose level) ) Decreases and is kept at a constant concentration. In addition, the blood glucose level lowered by exercise or the like is converted from glycogen to glucose, which is released into the blood to maintain a constant blood glucose level.

  The blood glucose level regulation mechanism involves incretin hormone (hereinafter simply referred to as incretin) secreted into the blood from the intestinal tract when the postprandial blood glucose level is high. Incretin consists of two types of peptides, one is glucagon-like peptide-1 (GLP-1) secreted from L cells present in the ileum and colon, and the other is glucose secreted from K cells in the duodenum. Dependent insulin secretion stimulating polypeptide (GIP). These peptides act on the pancreatic β-cells when the blood concentration rises as a result of ingestion of the meal, promote insulin secretion, and are not secreted more than necessary when the blood glucose level is within the normal range. It also works to lower blood sugar levels by suppressing the secretion of glucagon, which has the effect of releasing stored energy.

  Incretin also works outside the pancreas, and GLP-1 works on the central nervous system and digestive tract (mainly the stomach) to suppress appetite and suppress gastrointestinal motility. Not work. GIP also works on fat cells and brown fat cells to promote glucose uptake and eliminate glucose from the blood.

  On the other hand, incretin is known to be rapidly degraded after its secretion by DPP (dipeptidyl peptidase) IV, its degrading enzyme, and lose its in vivo activity. Also requires inhibition of DPPIV. In recent years, antidiabetic drugs having a mechanism of action related to incretin have been developed. There is a GLP-1 receptor agonist as a type that promotes GLP-1 production. As a feature of the GLP-1 receptor agonist, the structure of GLP-1 itself is changed to make it difficult to be decomposed by DPPIV. Examples of such drugs include liraglutide and exenatide (for example, Patent Document 1). Moreover, there exists a DPPIV inhibitor as a type which prevents decomposition | disassembly of produced GLP-1 from the effect | action of DPPIV, and there exist a vildagliptin, sitagliptin, alogliptin, etc. as a chemical | medical agent (for example, patent document 2).

  In addition to lowering blood glucose, DPPIV inhibitors have the effect of not lowering fasting blood glucose too much, and conventional therapeutic drugs for diabetic patients do not take food or anti-diabetic drugs are too effective Due to these factors, the difficulty of controlling blood glucose level has been overcome, and it is regarded as a new therapeutic agent for patients with type II diabetes.

  In the above-mentioned mechanism of glucose metabolism, there is an intestinal absorption process of glucose. In this process, the glucose concentration in blood is controlled by a transporter (SGLT-1) on small intestinal epithelial cells. SGLT-1 is mainly distributed in the intestinal epithelium and is also present in the proximal tubule, trachea, heart, etc. of the kidney. As its function, it absorbs glucose in the intestine and reabsorbs glucose in the kidney. Note that SGLT-2 present in the kidney only reabsorbs glucose in the kidney. SGLT-1 and SGLT-2 have the effect | action which absorbs glucose from a digestive tract and suppresses glucose excretion in urine. When the supply balance of glucose is disrupted, the function of SGLT-1 in the heart is expanded and the patient is likely to suffer from cardiac dysfunction. In particular, patients with type II diabetes have a concern of causing diseases such as cardiomyopathy and myocardial ischemia ( Non-patent document 1).

  Acarbose, voglyse, miglitol and the like are known as pharmaceuticals based on the glucose absorption inhibitory action. These are drugs involved in intestinal absorption, which inhibits an enzyme (α-glucosidase) that decomposes disaccharides present in the intestinal tract into monosaccharides and has a function of relaxing digestive absorption of glucose. Examples of pharmaceuticals based on SGLT-1 inhibition and SGLT-2 inhibition include dapagliflozin, empagliflozin, canagliflozin and the like, and these are drugs having glucose reabsorption inhibitory action and excretion action (for example, Patent Document 3).

  If glucose metabolism, especially glucose metabolism, does not operate normally, it induces not only obesity and diabetes but also glycation in vivo. Glication is a reaction in which excess sugar in the body is linked to protein, and is a non-enzymatic reaction between an amino group of a protein constituting a living body and a carbonyl group of a reducing sugar such as glucose. The glycation proceeds, and glycation end products (AGEs) are formed through various chemical reactions such as oxidation, dehydration, and condensation. AGEs are powerful aging-promoting substances, and when accumulated in the body, tissues (organs and bones) and blood vessels constituting the body become brittle, and the tissues in the body become stiffened and fossilized. In addition, sagging and wrinkles can occur in the skin, the elasticity of blood vessels is lost, arteriosclerosis is promoted, and there is a possibility of leading to diseases such as osteoporosis, cataracts, and Alzheimer's dementia.

  In addition, as a characteristic of highly obese people, visceral fat binds to cellular receptors and activates inflammatory signals, causing inflammation in the cells. Enlarged visceral fat increases TNF-α and IL-6, which are proinflammatory cytokines, or M1 macrophages, which are pro-inflammatory macrophages, and simultaneously secretes proinflammatory adipocytokines such as the chemokine MCP-1. Reduce the secretion of adiponectin, which shows anti-inflammatory. In addition, the immune cells involved in adipocytes are not limited to macrophages, and expression of inflammatory reactive immune cells such as cytotoxic T cells is induced. In addition, stimulation with these proinflammatory cytokines suppresses insulin stimulation signals derived from insulin receptors. That is, insulin resistance in which the effectiveness of insulin is deteriorated, insulin does not work in the living body, blood sugar level cannot be controlled, and serious diseases such as diabetes may be caused (Non-patent Document 2). .

The button bow (scientific name: Peucedanum japonicum ) is a large perennial of the genus Amaranthaceae. It is also known as a butterfly (long-life grass) or sacna in other names, and its distribution is vegetated from the subtropics of East Asia to the temperate zone. In Japan, it grows naturally in Okinawa from the southern part of Kyushu, and grows under severe natural conditions such as the sea breeze and the shining sun. The plant height is about 80cm, the root leaves are divided into two wings, resembling the leaves of a button, and in summer, white florets grow densely on the branches. Leaves and stems are edible as vegetables, and typical cooking methods include tempura, green juice, and tea. Nutritional value is rich in vitamins A, B2, B6, E, β-carotene, calcium, fiber and the like.

  In addition, it is said that buttonbow is good for high blood pressure, arteriosclerosis, rheumatism, neuralgia, asthma, whooping cough, cold, fever, etc., especially the root has been a substitute for medicinal ginseng as a nourishing tonic. . In recent years, attention has been focused on antioxidants that scavenge active oxygen, substances that inhibit disaccharide-degrading enzymes, antioxidant effects and melanin production-suppressing effects, and proposals for use as skin external preparations that are useful for preventing photoaging ( Patent Documents 4, 5, and 6).

Table 2011/052523 Special table 2009-508955 Special table 2012-523374 gazette JP 2004-026697 A Japanese Patent No. 4269036 JP 2004-359732 A

  Banerjee SK. Cardiovascular Research, 84 (1), 111-118, 2009   Ayaka Ito et al., Adiposcience, Volume 6 (1), pp. 16-21, 2009

  In daily meals, it is a fact that glucose-containing foods and drinks containing glucose, glucose fructose liquid sugar, etc. are customarily ingested in various amounts and in large quantities. Therefore, it cannot be said that the active ingredient that expresses the substance is effective enough for an actual meal, sugar metabolism, and sugar absorption. In addition, it has not been known in the past that button bow fu promotes GLP-1 production, inhibits DPPIV activity, or inhibits glucose absorption.

As described above, in vivo GLP-1 lacks stability and is degraded and inactivated by DPPIV. Therefore, even if ingested or administered from outside the body, it is difficult to maintain its activity in the body. Conventional drugs with 1-receptor action are likely to cause side effects such as constipation, nausea / vomiting, and diarrhea because they act on the gastrointestinal tract, and when used in combination with other antidiabetic drugs, tremors, dizziness, facial pallor, etc. Attention has been drawn to the fact that symptoms of hypoglycemia are likely to appear, and the development of components and substances derived from natural materials having an action of promoting GLP-1 production in vivo has been demanded.
In addition, conventional drugs having DPPIV inhibitory action have long been noted for renal and hepatic disorders in their metabolic and excretory pathways, but DPPIV inhibitors have many extrapancreatic effects (lowering blood glucose levels). Actions other than insulin secretion), for example, side effects such as increased cardiac output in the cardiovascular system and glucose metabolism in skeletal muscles are pointed out. There are also concerns.
In addition, drugs based on the glucose absorption inhibitory action reduce the postprandial blood glucose peak and are effective when taken with meals. However, hyperglycemic symptoms caused by factors other than meals (for example, dyslipidemia and preparatory type II diabetes) The prevention and improvement of the army were not fully satisfactory. Furthermore, these drugs are subject to restrictions such as needing to be administered under the guidance and management of a doctor, and are difficult to use for obese people who do not have type II diabetes and those with diabetes reserves. Was the current situation.
Therefore, in the present invention, a GLP-1 production promoter, a DPPIV inhibitor, and a glucose absorption inhibitor that can be easily incorporated into daily eating habits, are safe, and can be applied without selecting a user are developed. Furthermore, another object of the present invention is to provide an industrial aspect in which the agent can be used effectively.

  In order to solve the above-mentioned problems, the present inventors have conducted extensive studies on a wide variety of materials for promoting GLP-1 production in the living body and suppressing or inhibiting DPPIV activity and glucose absorption ability. When a plant of this kind is used, an extremely high GLP-1 production promoting action, DPPIV inhibiting action and glucose absorption inhibiting action are expressed, and this is applied to industrial fields such as foods and drinks, pharmaceuticals, quasi drugs, cosmetics, animal feeds, etc. The present invention has been completed.

That is, the features of the present invention are as follows.
(1) A GLP-1 production promoter comprising button pea ( Pucedanum Japanicum ) as an active ingredient.
(2) A DPPIV inhibitor comprising button pea ( Pucedanum Japanicum ) as an active ingredient.
(3) A glucose absorption inhibitor containing button pea ( Pucedanum japonicum ) as an active ingredient.
(4) The agent according to any one of the above (1) to (3), wherein the active ingredient is an extract obtained by treating button pea ( Pucedanum japonicum ) with an extraction solvent comprising water and / or a lower monohydric alcohol. .
(5) The agent according to (4), wherein the extraction solvent is water-containing ethanol (water content: about 1% by mass to about 50% by mass).
(6) An oral composition comprising at least one agent according to any one of (1) to (5).
(7) The composition according to the above (6), which is for suppressing an increase in blood glucose level.
(8) The composition according to the above (6), which is for reducing the amount of meal and / or weight.
(9) The composition according to any one of (6) to (8), wherein the aspect is a pharmaceutical product, quasi drug, food or drink, food additive, pet food, or feed.

  The agent of the present invention comprises button fountain as an active ingredient, and has an effect of enhancing in vivo production of intestinal secretory hormone (incretin), particularly GLP-1, involved in sugar metabolism. It can be used as an anti-obesity measure in terms of contributing to a decrease in value, appetite suppression and digestive tract (stomach) movement suppression. In addition, in order to exert the effect of inhibiting the activity of DPPIV that degrades the incretin, the blood incretin function is maintained, the insulin secretion is promoted, the glucagon secretion is suppressed, and the blood glucose level is maintained in the normal range. It becomes possible. Furthermore, since the effect of inhibiting glucose absorption from the small intestinal tract is exerted, it is possible to reduce the blood glucose concentration and suppress an increase in blood glucose level. Such an effect becomes more remarkable when the active ingredient is a hydrous ethanol extract. The agent of the present invention can be taken together with daily meals, normalizing the glucose metabolism function of the living body, obesity, diabetes, tissue sclerosis induced by in vivo glycation, vascular weakening, It is useful in preventing, ameliorating or treating aging-related diseases such as skin disorders, arteriosclerosis, osteoporosis, and cataracts. For this purpose, pharmaceuticals, quasi drugs, food and drink, food additives, pet food, feed, etc. It can be used effectively in the form.

  Next, modes for carrying out the present invention will be described in detail.

The agent of the present invention is characterized in that button bow fu ( Peucedanum japonicum ) is used as an active ingredient. As the raw material for the button bow, what is native or cultivated in the area from the west of central Honshu to Okinawa, the southern part of the Korean Peninsula, China, the Philippines, etc. may be used as appropriate. Any part of the plant body (leaves, stems, roots, etc.) can be used, and this dry powder or an extract treated with water and / or a hydrophilic organic solvent is used as a substantial active ingredient of the agent. To do.

  The button-bow-fue according to the present invention can be produced by processing it into a dry powder by a known method or extracting it by any method. In order to produce an extract, it is preferable to perform an extraction treatment using water and / or a lower alcohol. The extraction treatment method may be performed under normal temperature to pressurization (atmospheric pressure + 1 to 3 atm) and normal temperature to the boiling point of the solvent according to the solvent used. As the extraction solvent, a known solvent used for producing a general plant extract may be appropriately selected and used. For example, water (hot water), lower alcohols such as methanol, ethanol, absolute ethanol, butanol, acetone, etc. , Diethyl ether, ethyl acetate, a mixed solvent thereof and the like. In the present invention, it is more preferable to use water and / or lower monohydric alcohol, water and / or ethanol are more preferable, and water-containing ethanol (water content: about 1% by mass to about 50% by mass) is most preferable. When the extraction solvent is water-containing ethanol, more specifically, when the agent of the present invention is a GLP-1 production promoter, the water content is about 1% to about 50% by weight, and when the agent is a DPPIV inhibitor, the water content is: About 30% by mass to about 50% by mass, and in the case of a glucose absorption inhibitor, the water content is preferably about 1% by mass to about 50% by mass. When the moisture content is out of the above range, the desired effect of the present invention tends to be lowered.

When the agent of the present invention is used for eating and drinking, the raw material is a button fountain leaf, stem, root part or a mixture thereof, which is dried by a conventional method or a pulverized product thereof, or the solvent From the viewpoint of production cost and convenience, it is preferable to use an extract obtained by extraction with the above, a concentrate thereof, a dried product or a powder thereof.
In addition, when used for pharmaceutical purposes, the extract, concentrate, extract extracted using the above-mentioned solvent or other hydrophilic organic solvent, or these are further collected for processing such as solvent fractionation or adsorbent fractionation. A purified product with high purity is desirable.

  The dried product, extract, refined product, etc. of button fountain obtained as described above are in the form of a solid, paste or liquid, and these may be used as they are as the agent of the present invention. It is also possible to use a known additive in a use for which is used in combination as necessary, and to contain it by a conventional method to prepare a composition. Here, known additives are preferably those usually used for oral ingestion and those usually used as external preparations. For example, excipients, binders, disintegrants, lubricants, wetting agents, fluids It is possible to use additive substances such as agents, preservatives, surfactants, stabilizers, diluents, solubilizers, tonicity agents, bactericides, preservatives, corrigents, flavoring agents, coloring agents, and fragrances. It is.

  In the present invention, the above-mentioned agents can be used as a part of blended raw materials of various products in food and drink, food additives, pharmaceuticals, quasi drugs, feeds, and other industrial fields. In particular, an oral composition is preferably used in order to sufficiently exhibit GLP-1 production promoting action, DPPIV inhibitory action and / or glucose absorption inhibitory action, and preferred embodiments of this oral composition are foods, drinks, pharmaceuticals or pharmaceuticals. It is an outside product. Examples of these are described below, but the present invention is not limited thereby. In the case of a composition in which a plurality of the above-mentioned three kinds of agents are blended, the ratio of each agent can be appropriately set according to the expected action / effect.

  The form of the oral composition containing the agent of the present invention can be an oral preparation such as a granule, a tablet, a capsule or a liquid. The content of button fountain in such a pharmaceutical composition is difficult to define uniformly depending on the type and content of the raw materials used together, but in the case of the extract, it is generally about 0.01% to 90% by weight, more preferably About 0.1% by mass to about 70% by mass. When the content is less than about 0.01% by mass, the desired effect of the present invention is not recognized, and when it exceeds about 90% by mass, it is difficult to prepare a practical pharmaceutical composition. In the case of other than the above extract (non-extracted product or purified product), it may be calculated in consideration of the extraction yield and the purification concentration rate. The composition of the present invention is preferably used orally taken or administered. A measure of the amount to take or administer the oral composition of the present invention is about 0.5 g to about 10 g, preferably about 0.05 g to about 0, per day for a human adult, based on the extract contained in the composition. .1 g.

  Specific examples of food and drink include beverages such as vegetable juice, fruit juice drinks, soft drinks, tea, instant noodles, soup, jelly, pudding, yogurt, cake premix products, confectionery, sprinkles, miso, soy sauce, sauce, dressing , Mayonnaise, vegetable cream, seasonings such as grilled meat sauce and noodle soup, noodles, udon, soba noodles, spaghetti, ham and sausage and other processed meat and fish products, hamburger, croquette, boiled, jam, milk, cream, butter, In addition to various processed foods such as powders such as spreads and cheese, solid or liquid dairy products, margarine, bread, cakes, cookies, chocolate, candy, gummi, gum, etc., powders, granules, pills, Tablet, soft capsule, hard capsule, paste or liquid dietary supplement, food for specified health use, functional food Mention may be made of health food products, the concentrated liquid diet and dysphagia for food diet and the like.

  When used as a pharmaceutical or quasi-drug, together with the above-mentioned agents, pharmaceutically acceptable known excipients and additive substances are used in combination without departing from the spirit of the present invention, and processed by a conventional method to produce tablets, Prepare capsules, granules, powders, liquids, etc. In order to suppress the increase of blood glucose level in vivo for the prevention, improvement and treatment of obesity based on the in vivo GLP-1 production promoting effect, DPPIV inhibitory effect and / or glucose absorption inhibiting effect. It can be applied for the prevention, improvement and treatment of the various diseases and symptoms related to abnormal sugar metabolism. In addition, the content and dosage of the agent blended in these drugs and quasi-drugs are the same as in the case of the oral composition described above.

  Further, in order to use the agent as pet food or livestock feed, the agent is blended with various known feeds or drinking water as in the case of the above-mentioned food and drink, and is tableted together with known raw materials and additive substances. It can be processed into a granular form, a capsule form, and the like. The content and intake of the agent in these feeds are almost the same as in the case of the oral composition described above.

  EXAMPLES Next, although an Example is given and this invention is demonstrated further in detail, this invention is not limited at all by these. In each example,%, part, and ratio are all based on mass.

Production Example 1
The dried powder of the button bow (from Kikaijima, Kagoshima Prefecture) is dried with a steam dryer, the dried product is crushed to a size of about 3 mm square or less, and this is further processed with a pulverizer and sieved. 1) was produced.

Production Example 2
Water (800 mL) was added to 100 g of button-powder fine powder obtained by the method described in Production Example 1, heated at 80 to 90 ° C. for 60 minutes, cooled to room temperature, and filtered to separate the filtrate. 600 mL of water was again added to the filtration residue and heated in the same manner. After cooling, the filtrate was collected by filtration. Both filtrates were combined, concentrated under reduced pressure, lyophilized and ground to produce 40.2 g of extract (sample 2).

Production Example 3
80 g of water-containing ethanol (water content: 45%) was added to 10 g of the fine powder of button bow obtained by the method described in Production Example 1, heated to reflux at 75-80 ° C. for 1 hour, cooled to room temperature, filtered and filtrated. Separated. 60 mL of water-containing ethanol (water content: 45%) was added again to the filtration residue and heated in the same manner. After cooling, the filtrate was collected by filtration. Both filtrates were combined, concentrated under reduced pressure, lyophilized and ground to produce 3.8 g of extract (sample 3).

Production Example 4
80 g of water-containing ethanol (water content 30%) was added to 10 g of the fine powder of button-bow-fu obtained by the method described in Production Example 1, heated to reflux at 75-80 ° C. for 1 hour, cooled to room temperature, filtered. The filtrate was separated. To this filtration residue, 60 mL of hydrous ethanol (water content 30%) was added again and heated in the same manner. After cooling, the filtrate was collected by filtration. Both filtrates were combined, concentrated under reduced pressure, lyophilized and pulverized to produce 3.5 g of extract (sample 4).

Production Example 5
After adding 80 mL of 99% ethanol to 10 g of the fine powder of buttoned bowfish obtained by the method described in Production Example 1, the mixture was heated to reflux at 75-80 ° C. for 1 hour, cooled to room temperature, and filtered to separate the filtrate. To this filtration residue, 60 mL of 99% ethanol was added again and heated in the same manner. After cooling, the filtrate was collected by filtration. Both filtrates were combined, concentrated under reduced pressure, lyophilized and crushed to produce 2.16 g of extract (sample 5).

Comparative production example 1
The dried leaves of green tea produced in Shizuoka Prefecture were roughly chopped and further processed with a pulverizer to produce a fine powder (sample 6) that passed through a sieve.

Comparative production example 2
1 kg of dried leaves of green tea produced in Shizuoka Prefecture was roughly chopped, 3 L of water was added, and the mixture was appropriately stirred at 85 ° C. for 20 minutes under normal pressure, then cooled to room temperature and filtered to separate the filtrate. 2 L of water was again added to the filtration residue and heated in the same manner. After stirring and cooling, the filtrate was collected by filtration. Both filtrates were combined and concentrated under reduced pressure. The concentrated solution was subjected to column chromatography filled with a synthetic adsorbent Diaion (registered trademark) HP-20 (manufactured by Mitsubishi Chemical Corporation). After washing with 5 L of water, 5 L of water-containing ethanol (water content 30%) was passed through to collect a fraction soluble in the water-containing ethanol. This solution was concentrated under reduced pressure, spray-dried and pulverized to obtain 25 g of a powdery green tea extract (sample 7). As a result of HPLC analysis of this extract, the content of catechins (including epigallocatechin gallate as a main component, gallocatechin gallate, epicatechin gallate, epigallocatechin, catechin, epicatechin, etc.) is 90.2%, The in content was 0.1% and others.

Test Example 1: GLP-1 Production Promoting Action GLP-1 production action was tested using dry powder and extract (sample 1, sample 2, sample 3, sample 4 and sample 5) of button pea as samples. GLP-1 producing cells (manufactured by Sumisho Pharma International Co., Ltd., cell name “NCI-H716”) were pre-cultured in 10% FBS-RPMI1640 medium. A 96-well microplate coated with Poly-D-Lysine was seeded at 1 × 10 ⁵ cells / well and cultured in 10% FBS-DMEM high glucose medium for 2 days.
Dissolve each sample in HBSS to a final concentration of 1.25, 2.5, 5.0 mg / mL, incubate for 4 hours after addition, then collect the supernatant and centrifuge to use as a sample It was. All subsequent test procedures were performed according to the manual of the Total GLP-1-HS ELISA measurement kit (manufactured by Yauchihara Laboratory). Add 25 μL of the sample or standard solution and 100 μL of buffer to the antibody-coated measurement plate, shake at room temperature for 18 hours, remove the reaction solution, wash the well with the washing solution, and then remove the labeled antibody solution. Add 100 μL, shake for 1 hour, remove the reaction solution, wash the inside of the well with a washing solution, add 100 μL of SA-HRP solution, shake for 0.5 hour at room temperature in a light-shielded state, and then add 100 μL of enzyme substrate solution. The mixture was allowed to stand at room temperature for 0.5 hours in a light-shielded state. 100 μL of enzyme reaction stop solution was added, and the absorbance was measured at 450 nm. A standard curve was drawn with a standard product to determine the amount of GLP-1 production, and the value corrected with protein by the BCA method was calculated.

  The results are shown in Table 1. In the same table, the numerical value is shown as a relative value when the value of the case where no sample was added simultaneously was set to 1. Table 1 shows that GLP-1 production-enhancing action was observed in both the dried powder and extract of button peas according to the present invention, and particularly strong hydrous ethanol extracts (Sample 3, Sample 4, Sample 5). It was confirmed that the production promoting effect was expressed.

Test Example 2: DPPIV Inhibitory Action DPPIV inhibitory action was tested using dry powder and extract (sample 1 to sample 5) of button fountain as a specimen and green tea extract (sample 7) as a comparative specimen. All test procedures were performed according to the manual of the DPPIV activity measurement kit (AnaSpec, USA). Dilute each sample with DMSO aqueous solution (final concentration 0.2%) to 0.25, 0.5, 1, 2 mg / mL (final concentration), and add 10 μL of each sample solution and 40 μL of DPPIV solution to the assay plate. Added. Next, 50 μL of the substrate solution was added and mixed, and incubated at 37 ° C. for 30 minutes, and then the fluorescence intensity at excitation wavelength: 354 nm and fluorescence wavelength: 442 nm was measured. The DPPIV inhibition rate was calculated from the fluorescence intensity of the specimen with respect to the fluorescence intensity of the control to which no sample was added. The DPPIV inhibition rate was calculated from the following formula.
DPPIV inhibition rate (%) = {1− (control−sample) / (control−blank)} × 100

  The results are shown in Table 2. In the table, the numerical values are shown as relative values when the value of the control test carried out at the same time is taken as 100. From Table 2, it can be seen that DPPIV inhibitory action is observed in both the dry powder and extract of button fountain according to the present invention, and in particular, the water-containing ethanol extract (samples 3, 4, and 5) exhibits a strong DPPIV inhibitory action. confirmed.

Test Example 3: Glucose absorption inhibitory action by CACO-2 cells Dry powder and extract of button potato as sample (sample 1 to sample 5), Lingonberry extract powder (manufactured by BEIJNG GINKO GROUP, comparative sample 1) as a comparative sample, Using red wine extract R (manufactured by Medience, comparative sample 2), the glucose absorption inhibitory action by CACO-2 cells was tested.
1 × 10 ⁴ CACO-2 cells (manufactured by DS Pharma Biomedical Co., Ltd., trade name “CACO-2”) are seeded on an insert well for a 24-well microplate, and the electric resistance value becomes 250 Ω / cm ² or more. To 10% FBS-DMEM high glucose medium. Before adding the specimen, the Aical and Basal sides were replaced with HBSS and incubated for 30 minutes. Glucose (10 mg / mL) and a sample were dissolved in HBSS to a final concentration of 1.25, 2.5, and 5.0 mg / mL and added, and then the basal cell passage solution was collected every 30 minutes. The glucose concentration of the cell passage solution incubated for 0 to 1.5 hours was measured with glucose CII-Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). The glucose absorption inhibition rate was calculated from the glucose concentration when the sample was added to the glucose concentration (control) when the sample was not added. The glucose absorption inhibition rate was obtained from the following calculation formula.
Glucose absorption inhibition rate (%) = {1− (control−sample) / (control−blank)} × 100

  The results are shown in Table 3. In the table, the numerical values are shown as relative values when the value of the control test carried out at the same time is taken as 100. From Table 3, the glucose absorption inhibitory action is recognized in any of the dry powder and extract of the button fountain according to the present invention, particularly in the case of a water-containing ethanol extract (samples 3, 4, and 5). It was observed that the effect was exerted.

Test Example 4: Glucose Level Inhibition Test by Ingestion of Cooked Rice A single ingestion test was conducted on seven adult adults (30s: 3 men, 4 women) who had consented to the following test.
The subject refrained from eating and drinking other than water after 22:00 on the day before the measurement, and measured the fasting blood glucose level on the morning of the measurement day (8:30). In the first test, only water was ingested (control group), in the second test, a solution of 1 g of button-bow powder (sample 1) suspended in 25 mL of water (sample 1 group), and in the third test, green tea leaves A solution of 1 g of powder (sample 6) suspended in 25 mL of water was ingested (sample 6 group). 5 minutes after ingesting water and sample, 200 g of commercially available packaged rice “Sato no Gohan” (Sato Food Industry Co., Ltd.) is ingested with a small amount of water, and after ingestion of rice, 30 minutes, 60 minutes, 90 minutes, 120 minutes, 150 The blood glucose level was measured in minutes, and the effect of button-bow fue on postprandial blood glucose level was evaluated. In addition, the blood glucose level was measured using a blood sugar self-monitoring device (manufactured by Roche Diagnostics, Inc., “Accu Check Aviva” (registered trademark)). In the evaluation, a change diagram of blood glucose level versus time at the time of ingestion or non-ingestion of buttonfish was drawn, and the area under the blood concentration curve (AUC) in this change diagram was calculated.

  As a result, compared with the area under the blood concentration curve, the sample 1 group was significantly lower than the control group. Moreover, it became a low value also compared with the sample 6 group. Among them, the sample 1 group showed a lower blood glucose level (maximum blood glucose level) 30 minutes after the meal than the control group and the sample 6 group. From these results, it was confirmed that the button bow powder according to the present invention has an effect of suppressing an increase in blood glucose level after a meal.

Test Example 5: Human monitor test 20 adult female volunteers (30-55 years old, average age: 43.5 years old) who gave consent to the following test were given placebo (dextrin: 250 mg) for the first 4 weeks. A test was conducted in which sample was taken twice a day (control group), and sample 1 (250 mg / dose) was similarly taken after completion of the control intake (sample 1 group). At the end of the study, evaluation was conducted by questionnaire survey on changes in body weight and meal contents.

  As a result, in the sample 1 group, the body weight decreased: 13 people, and did not change: 7 people. Thirteen persons whose body weight changed had changes such as a decrease in the amount and number of snacks and a decrease in the amount of meals. From this result, it is suggested that the button bow fu according to the present invention is useful for improving weight loss and lifestyle (meal pattern).

Prototype Example 1: Hard Capsule The above Sample 1 was subjected to a capsule filling machine, and a gelatin-coated hard capsule preparation with an internal volume of 250 mg per grain was made by a conventional method. When a tablet (containing the button bow fu of Production Example 1) was orally ingested in a monitor test, findings were found that a decrease in body weight and a decrease in the amount of food were observed. Therefore, this tablet can be used as an orally ingestible dietary supplement, pharmaceutical or animal feed for the prevention of obesity.

Trial Production Example 2: Tablets Tablets were produced by tableting the raw materials shown below by conventional methods. Here, as the button bow extract, any one of the button bow extract (samples 2 to 4) obtained in Production Examples 2 to 4 was used. These tablets are all stable and easy to take, and can be used as dietary supplements and pharmaceuticals.
(Combination raw material) (Mass per tablet (mg))
1. Button bow 50
2. Dextrin 100
3. Potato starch 39
4). Microcrystalline cellulose 30
5. Synthetic aluminum silicate 30
6). Calcium stearate 1

Prototype Example 3: Nutritional Supplement 1 g of the sample 1, 400 mg of leucine, 250 mg of isoleucine, 200 mg of valine, 300 mg of arginine, mixed and sieved powder of these were packaged and prototyped of a dietary supplement. This dietary supplement can be used not only for humans but also for pet foods.

Prototype Example 4: Food (Noodles)
The sample 1 was 15 g, the buckwheat flour was 350 g, and the strong flour was 150 g. While these were mixed, they were sifted and mixed with 150 ml of water to make a buckwheat. This did not give a sense of incongruity to the flavor and texture compared to the commercial products.

Prototype Example 5: Soft Drinks and Drinks 500 mg or 100 mg of Sample 1 or Sample 3 was added to 100 mL of commercially available soft drinks and drinks and mixed thoroughly to make a drink. This beverage was not inferior in flavor and storage stability compared to the original soft drinks and drinks. This can be used as a soft drink or a drink.

  The agent of the present invention is used to prevent, ameliorate, or treat obesity and glucose metabolism-related diseases by promoting GLP-1 production, DPPIV inhibition, and / or glucose absorption inhibition in the fields of pharmaceuticals, foods and drinks, feeds, and the like. Effective use for products.

Claims (2)

GLP-1 production promoter (excluding treatment for diabetes) comprising as an active ingredient an extract obtained by treating leaves of buttoned pea (Peucedanum japonicum) with water-containing ethanol (water content: 30% to 45% by weight). method of manufacturing a). Manufactured a DPPIV inhibitor (except for therapeutic use for diabetes) containing as an active ingredient an extract obtained by treating leaves of Peucedanum japonicum with water-containing ethanol (water content: 30% to 45% by weight). How to do .