JP2006335741A - Peroxisome proliferating agent-responsive receptor ligand composition derived from plant belonging to genus amorphophallus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a composition for preventing or treating insulin resistance or a life style-related disease such as metabolic syndrome, obesity, adiposis, diabetes, hyperlipidemia, hypertension or gout that is highly safe and easy to manufacture and can be used for foods and drinks such as health foods or health functional foods (foods for specific health care or nutrient functional foods), pharmaceuticals, quasi-drugs, cosmetics or the like. <P>SOLUTION: The composition for preventing or treating insulin resistance or the life style-related disease such as metabolic syndrome, obesity, adiposis, diabetes, hyperlipidemia, hypertension or gout comprises konnyaku extracts exhibiting peroxisome proliferating agent-responsive receptor ligand activities. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a peroxisome proliferator-responsive receptor ligand that can be used in foods and beverages such as health foods and health functional foods (specific health foods, functional nutrition foods), pharmaceuticals, quasi drugs, cosmetics, and the like. And a composition for preventing or treating lifestyle-related diseases such as insulin resistance or metabolic syndrome, obesity, obesity, diabetes, hyperlipidemia, hypertension, gout, etc., comprising the agent About.

Konjac ( Amorphophallus konjac K. Koch ) is a tropical Asian native plant of the Araceae family, and the corm part of the corm part has long been edible in Japan. The corms contain konjac mannan consisting of glucose and mannose. If the koji is crushed as it is, or if the koji is sliced and dried to a fine powder, and a large amount of water is added, it will absorb water and become a paste. Features. The edible konjac is the one that has been boiled and coagulated with lime water. The method of making directly from koji is still in progress and is highly valued in terms of flavor and color. However, in terms of storage and mass productivity, most konjacs in the market are refined refined products that contain a lot of glucomannan. It is made from flour. The dried and roughly crushed sliced konjac koji is called coarse powder, and the finer particles with a high density and heavy specific gravity are called fine powder. Light fine particle powder generated in the process of making fine powder from coarse powder is called flying powder, and the production ratio of fine powder and flying powder is almost equal. In the fine powder production process used in the efficient konjac production method, flying powder is removed, which is very fine compared to fine powder containing a lot of konjac mannan. However, it is speculated that the reason why the konjak produced from the whole rice cake has a good flavor is due to this flying powder component, and if this flying powder is used effectively, the whole konjac can be used. Konjac is a traditional Japanese food that is actively consumed as a low-calorie food-rich food. The efficacy of konjac currently said is due to konjac mannan and dietary fiber as the main components contained in konjac fine powder, lipid metabolism improving effect by suppressing liver lipid amount (Non-patent Document 1), and intragastric retention time The effect of improving glucose tolerance due to the prolongation action and glucose diffusion inhibition (Non-patent Document 2) has been reported. Flying powder is produced as a by-product of konjac by its mass production method, but its application in the form of human consumption has not been made at all due to its delicious taste, and there has been almost no report on ingredients and efficacy. In other words, konjac fine powder is increasingly used, while flying powder is highly nutritious with a high sugar content and protein content, but it is currently present due to the bitter taste and raw odor of oxalic acid, homogentisic acid, and phenolic substances. Is used only for fertilizer and feed. In addition, reports on flying powder and its extract are related to the production and utilization of glycosphingolipids from flying powder (Patent Documents 1 to 3), and reports on antihypertensive substances (Non-Patent Document 3). It is not known that the extract has peroxisome proliferator-responsive receptor ligand activity.

  Peroxisome proliferator-activated receptor (PPAR) is a ligand-dependent transcriptional regulator belonging to the nuclear receptor family responsible for controlling the expression of genes that maintain lipid and sugar metabolism. In mammals, PPARα, PPARδ (PPARβ, NUC-1, FAAR), and PPARγ are known to exist in three subtypes. PPARα is expressed in liver, kidney, skeletal muscle, etc., which have a high degree of fatty acid catabolism. In particular, high expression is observed in the liver. PPARδ is expressed universally in tissues. PPARγ has two isoforms, PPARγ1 and PPARγ2, PPARγ1 is expressed in immune system organs, adrenal gland and small intestine in addition to adipose tissue, and PPARγ2 is specifically expressed in adipose tissue ( Non-patent document 4).

PPARγ ligands include unsaturated fatty acids such as α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, arachidonic acid metabolites such as 15-deoxy Δ ^12,14 -prostaglandin J2 and Δ ¹² -prostaglandin J2, Eicosanoids such as 9-hydroxyoctadecadienoic acid and 13-hydroxyoctadecadienoic acid are known, and conjugated unsaturated fatty acids having 10 to 26 carbon atoms having a conjugated triene structure or a conjugated tetraene structure are PPARγ ligands, Among synthetic compounds, thiazolidine derivatives such as troglitazone, pioglitazone, and rosiglitazone are known to be PPARγ ligands (Non-patent Document 4).

  In addition to the above compounds, monoacylglycerol (Patent Document 4), gallic acid esters, galloyl tannins, polyphenols such as quercetin, flavone, isoflavone, catechin, epicatechin (Patent Document 5) and the like are PPAR ligands. It is known that

PPARα ligand mainly activates liver PPARα and enhances lipid metabolism and sugar metabolism, thereby improving insulin resistance and lowering blood lipid and blood glucose levels (Non-patent Document) 5).
PPARγ ligand activates PPARγ in adipocytes, increases small adipocytes with normal function differentiated from preadipocytes, and increases production and secretion of TNFα and free fatty acids that induce insulin resistance Insulin resistance is improved by reducing hypertrophic fat cells by apoptosis, and has a blood glucose level lowering action, etc. (Non-patent Document 6).

Thus, PPAR ligands are expected to improve insulin resistance and to prevent and / or improve lifestyle diseases such as hyperlipidemia, diabetes and obesity. However, the drugs shown above are expensive and have disadvantages such as side effects due to long-term administration. In addition, polyacylphenols such as monoacylglycerol, gallic acid esters, galloyl tannins, quercetin, flavones, isoflavones, catechins and epicatechins have disadvantages such as insufficient PPAR ligand action. Therefore, a material having high safety and high PPAR ligand action is desired.
JP2002-38183 JP2003-2835 JP2004-168738 JP2001-354558 JP2002-80362 Nutrition Journal, Vol. 31, 152, (1973) Nutr. Rep. Int., 23, 1145, (1981) Medical Research, 46, 22, (1976) Journal of Medicinal Chemistry, 43, 527-550, (2000) Diabetes, 50, 411, (2001) Ayumi of Medicine, 198, 747, (2001)

  INDUSTRIAL APPLICABILITY The present invention is highly safe and can be easily manufactured, and is used for foods and drinks such as health foods and health functional foods (specific health foods, nutritional functional foods), pharmaceuticals, quasi drugs, cosmetics, and the like. Peroxisome proliferator-responsive receptor ligand agent, or a composition containing the same, and insulin resistance, or metabolic syndrome, obesity, obesity, diabetes, hyperlipidemia, hypertension, gout, etc. It is an object to provide a composition for preventing or treating lifestyle-related diseases.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have found peroxisome proliferator-responsive receptor ligand activity in konjac extract, leading to the present invention.

  That is, the present invention provides the following.

(1) A peroxisome proliferator-responsive receptor ligand composition comprising a konjac plant ( Amorphophallus ) material as an active ingredient. Preferably, a peroxisome proliferator-responsive receptor ligand composition comprising an organic solvent extract of konjac plant ( Amorphophallus ) material as an active ingredient.

  (2) A composition for preventing or treating insulin resistance, or a composition for preventing or treating lifestyle-related diseases, comprising the peroxisome proliferator-responsive receptor ligand composition. Preferably, the composition wherein the lifestyle-related disease is metabolic syndrome or at least one disease selected from the group consisting of obesity, obesity, diabetes, hyperlipidemia, hypertension, and gout.

  (3) A food / beverage product, cosmetic product, or pharmaceutical product comprising the peroxisome proliferator-responsive receptor ligand composition.

  (4) A method for preventing or treating insulin resistance or lifestyle-related diseases using the peroxisome proliferator-responsive receptor ligand composition.

(5) A method for producing a peroxisome proliferator-responsive receptor ligand composition, comprising extracting a konjak plant ( Amorphophallus ) material using an organic solvent.

  According to the present invention, a composition for preventing or treating a lifestyle-related disease derived from a natural product can be provided. In addition, a peroxisome proliferator-responsive receptor ligand agent and a composition for preventing or treating lifestyle-related diseases containing the same as an active ingredient can be provided. The present invention is used for foods and drinks or pharmaceuticals, quasi drugs, cosmetics, etc., so that lifestyle resistance such as insulin resistance, metabolic syndrome, obesity, obesity, diabetes, hyperlipidemia, hypertension, gout, etc. It is useful as a composition for preventing or treating diseases.

The composition for preventing or treating lifestyle-related diseases of the present specification comprises an konjac plant material such as a konjac plant or a processed product thereof as an active ingredient, and contains an effective amount within a range in which the effect of preventing or treating lifestyle-related diseases can be exhibited. It only has to be.
In addition, the peroxisome proliferator-responsive receptor ligand composition of the present specification only needs to contain a konjak plant material as an active ingredient and an effective amount within a range in which peroxisome proliferator-responsive receptor ligand activity can be exerted.

Examples of the konjak genus plant include Amorphophallus konjac , Amorphophallus bulbifer , Amorphophallus campanulatus, etc., preferably Amorphophallus konjac species grown in large quantities in Japan, and more preferably Amorphophallus konjac K. Koch (konjac ).

  In addition, the collection time, the growing year, the cultivation method, and the cultivation period of the konjac plant used in the present invention are not particularly limited, and the konjac plant material is preferably a processed product derived from corms, and konjac flour, particularly konjac. Flying powder is preferred.

  The composition derived from konjac plant material used in the present invention can be obtained, for example, by solvent extraction from konjac powder. The extract can be used in the present invention as an extract, or as a crude extract or semi-purified extract as long as it does not contain impurities inappropriate for beverages and pharmaceuticals.

As the extraction solvent, polar solvents such as alcohols (eg, ethanol, lower alcohols of methanol or polyhydric alcohols such as glycerin), acetic acid, etc., alone or in any combination of two or more liquids, or a mixture with water Can be used as Among them, a water-soluble organic solvent is preferable, a lower alcohol having 1 to 3 carbon atoms is more preferable, and ethanol is particularly preferable from the viewpoint of safety of the residual solvent. The concentration of the water-soluble organic solvent is preferably an aqueous solution of 10% by volume or more, more preferably 50% by volume or more, and particularly preferably 70% by volume. Moreover, it can be used even if it combines each solvent extract. The amount of the extraction solvent to be used is preferably 0.5 to 50% by weight of the konjac plant material to be used, and more preferably 2 to 10% by weight.
In the case of performing solvent extraction, for example, fine powdered powder is usually immersed in 1 to 20 times the amount of extraction solvent, extracted by stirring and / or standing, and an extract can be obtained by filtration or centrifugation. Next, the extract can be obtained by removing the solvent and water content from the obtained extract by concentration and / or concentration to dryness, and further freeze drying. Extraction temperature is not specifically limited, Generally it can implement suitably at -20-121 degreeC, 1-80 degreeC of normal room temperature, Preferably it is 20-60 degreeC. The extraction time is not particularly limited, and it can be suitably carried out usually from 0.1 hour to 1 month, preferably from 0.5 hour to 7 days. After extraction, it may be in the form of a solution, paste, gel, or powder by concentration to dryness or oil or fat. Depending on the case, extraction under conditions of 30 to 120 ° C. or extraction under supercritical conditions such as autoclave or carbon dioxide is possible, and the extraction time may be set appropriately. If necessary, it can be further purified by an arbitrary operation using an activated carbon column, an ion exchange resin, or the like.

  As one embodiment of the present invention, an extract obtained by extracting konjac plant material with a 70% aqueous ethanol solution can be used. The amount of 70% ethanol aqueous solution used for extraction is preferably an amount capable of sufficiently dissolving the active ingredient contained in the konjac plant material.

  The composition comprising the konjak plant material in the present invention as an active ingredient is a work process for drying and roughly crushing sliced konjac koji, and making a fine powder mainly composed of glucomannan having a high density and a high specific gravity. Can be used. It is also possible to use a konjac plant that has been collected and washed to remove dirt and organisms adhering to the surface and then crushed by cutting, ball milling, ultrasonic treatment, homogenizer, or the like. Prior to crushing, air drying or freeze drying may be performed. In the case of solvent extraction, it is preferable that the powdered or crushed konjac plant material is repeatedly extracted several times with an extraction solvent while stirring.

  The composition containing the konjac plant material of the present invention as an active ingredient can be adjusted to an appropriate concentration by diluting or concentrating as necessary. For example, the composition of the present invention can be mixed with fats and oils to make a cream. Furthermore, a powdery lifestyle-related disease prevention-improving composition can be obtained by spray-drying a solution or an extract containing the composition. Thus, the concentration and shape of the composition of the present invention are not particularly limited, and can be appropriately determined according to the application.

  The composition of the present invention may contain components other than the konjac plant material. Since the composition of the present invention contains konjac plant material used for food as an active ingredient, it is highly safe for living bodies. For this reason, the extract of the present invention can be contained in a wide range of products such as pharmaceuticals, functional foods, foods and drinks, quasi drugs, cosmetics, and bath preparations.

  Since the component of the genus plant material of the present invention or the extract of the konjac plant material has a peroxisome proliferator-responsive receptor ligand, insulin resistance, or metabolic syndrome, obesity, obesity, diabetes, hyperlipidemia It is useful as a composition for preventing or treating lifestyle-related diseases such as hypertension and gout.

  The lifestyle-related diseases referred to here are based on the opinion statement “Basic direction of disease countermeasures focusing on lifestyle” published by the Public Health Council (December 18, 1996). It refers to a group of diseases in which lifestyle habits such as eating habits, exercise habits, rest, smoking, and drinking are related to the onset and progression. Lifestyle-related diseases include metabolic syndrome, obesity, obesity, diabetes, hyperlipidemia, hypertension, gout, high body fat percentage, hyperglycemia, hyperinsulinemia, hypercholesterolemia, hypertriglyceridemia, hypertension , And hyperuricemia. In lifestyle-related diseases, these are not a single disease but are related to each other.

  In the present invention, insulin resistance refers to a state where the absorption promotion action of sugar, which is the main action of insulin, is weak in liver, adipocyte, and skeletal muscle. Prevention of insulin resistance refers to preventing or delaying deterioration of a value that is an index of insulin resistance by the SSPG (steady-state plasma glucose) method or the like. Improvement of insulin resistance refers to improvement of the above-mentioned value that serves as an index of insulin resistance.

  In the present invention, prevention of metabolic syndrome is defined by the Japan Society of Internal Medicine and the like in the Journal of Japan Society of Internal Medicine in April 2005, the condition based on the diagnostic criteria or the borderline condition, the US Hyperlipidemia Treatment Guidelines (NCEP-ATPIII ) To prevent or delay the state or boundary state according to the definition and diagnostic criteria published in 2001, or the state or boundary state according to the definition and diagnostic criteria published by the WHO in 2001. Further, improvement of metabolic syndrome refers to bringing the above-mentioned hyperlipidemia state or boundary region state closer to the state defined as the normal region in the above guidelines. The metabolic syndrome referred to here is synonymous with “visceral fat syndrome” and multiple risk factor syndrome as described by Matsuzawa et al. (Matsuzawa et al., Diabetes / Metabolism Reviews, 13, 3-13, 1997).

  In the present invention, prevention of hyperlipidemia refers to the state of hyperlipidemia or borderline condition defined by the Japanese Society for Arteriosclerosis in the Guidelines for the Treatment of Arteriosclerotic Diseases (issued in September 2002). Refers to preventing or delaying. In addition, improvement of hyperlipidemia refers to bringing the state of hyperlipidemia or boundary region described above closer to the state defined as the normal region in the above guidelines.

  In the present invention, diabetes prevention refers to preventing or delaying a state of diabetes or borderline defined by the Diabetes Society of Japan 2002-2003 (issued in May 2002). Point to. Moreover, improvement of diabetes refers to approaching the state defined as the normal region by the guide from the above-mentioned diabetes state or border region state.

  In the present invention, obesity prevention means that the Japanese Society of Obesity is in a state defined as obesity or obesity in the obesity / obesity instruction manual 2nd edition (issued in July 2001). It refers to preventing or delaying. The improvement of obesity refers to the approach that the above academic society defines as obesity or obesity to the state that the above academic society defines as a normal range.

  The PPAR ligand composition of the present invention and a composition for preventing or treating lifestyle-related diseases such as insulin resistance, metabolic syndrome, or obesity, obesity, diabetes, hyperlipidemia, hypertension, gout, etc. It can be used for eating and drinking and for medicine, and its form is not limited. For example, health foods (specific health foods, nutritional functional foods), health foods, dietary supplements such as nutritional supplements, OTC, etc. It can be used as a readily available medicine or quasi drug.

  The PPAR ligand composition referred to in the present invention is a substance containing a compound having the ability to bind to the PPAR ligand binding region, that is, PPAR ligand activity. The PPAR ligand composition of the present invention regulates insulin and metabolic syndrome, obesity, obesity, diabetes, by regulating lipid and sugar metabolism in tissues such as liver, adipocytes, and skeletal muscle via PPAR. Lifestyle-related diseases such as hyperlipidemia, hypertension and gout can be prevented or improved. That is, the PPAR ligand agent of the present invention positively or negatively expresses various genes related to lifestyle-related diseases such as insulin resistance, metabolic syndrome, obesity, obesity, diabetes, hyperlipidemia, hypertension and gout. Can be controlled. Examples of the various genes include acyl CoA oxidase, medium chain acyl CoA dehydrogenase, carnitine palmitoyltransferase, long chain acyl CoA synthase, fatty acid binding protein, lipoprotein lipase, apolipoprotein, and uncoupling protein. It is not limited to.

  The PPAR ligand activity can be determined by, for example, a reporter assay (Cell, 1995, 83, 803-812) that evaluates the binding of a PPAR ligand binding region to a fusion protein of GAL4 by the expression of luciferase, and the PPAR ligand binding region. It can be measured by a competition binding assay (Cell, 1995, 83, 813-819) using the contained protein. In these assays, the activity of the sample is generally compared to the solvent control, and a sample that exhibits significantly higher activity than the solvent control and is dose-dependent is evaluated as “with PPAR ligand activity”.

  The PPAR ligand agent of the present invention only needs to have a ligand activity for at least one PPAR among PPARα, γ, and δ.

  When used as a food or drink, it can be taken directly as it is, or it can be ingested in a form that is easy to take, such as capsules, tablets, granules, using additives such as known carriers and auxiliaries. be able to. The content of the peroxisome proliferator-responsive receptor ligand composition of the present invention in these preparations is not particularly limited, but is preferably 0.1 to 100% by weight, more preferably 2 to 95% by weight, and still more preferably 10 to 10%. 90% by weight. Furthermore, mixed with food and drink ingredients, confectionery such as chewing gum, chocolate, candy, jelly, biscuits and crackers; frozen confectionery such as ice cream and ice confectionery; beverages such as tea, soft drinks, energy drinks and beauty drinks; udon Noodles such as Chinese noodles, spaghetti and instant noodles; kneaded products such as salmon, bamboo rings and half pieces; seasonings such as dressings, mayonnaise and sauces; fats and oils such as margarine, butter and salad oil; bread, ham, soup, retort food, It can be used for all foods and drinks such as frozen foods. When the composition for preventive treatment of the present invention is ingested for eating and drinking, the amount of intake is preferably 0.01 to 1000 mg / kg, more preferably 0.1 to 100 mg / kg per adult day as the extract. It is weight.

When used as a pharmaceutical, the dosage form is not particularly limited, and examples thereof include capsules, tablets, granules, injections, suppositories, and patches. In formulation, other pharmaceutically acceptable formulation materials such as excipients, disintegrants, lubricants, binders, antioxidants, colorants, anti-aggregation agents, absorption enhancers, solubilizers An agent, a stabilizer and the like can be added as appropriate. Preferable examples of the binder include starch, trehalose, dextrin, gum arabic powder and the like. Preferable examples of the lubricant include stearic acid, talc, sucrose fatty acid ester, polyethylene glycol and the like. Preferable examples of the disintegrant include starch, carboxymethyl cellulose, corn starch and the like. Preferable examples of the stabilizer include fats and oils and propylene glycol. Preferable examples of the emulsifier include anionic surfactants, nonionic surfactants, polyvinyl alcohol and the like. Preferable examples of the buffer include buffers such as phosphate, carbonate and citrate.
The dosage of these preparations is preferably 0.01 to 1000 mg / kg body weight, more preferably 0.1 to 100 mg / kg body weight per adult day per day in terms of the extract, divided into 1 to several times. Administer.

  When used as a quasi-drug, other additives are added as necessary, for example, ointments, liniments, aerosols, creams, soaps, facial cleansers, whole body cleansers, lotions, lotions, baths It can be used for agents and the like, and can be used locally.

  EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.

Example 1 Preparation of Konjac Extract 10 g of fine powdered konjac flying powder (Okawa Co., Ltd.) was extracted with 100 ml of 70 vol% ethanol aqueous solution (room temperature (23-24 ° C.), left in the dark for 3 days. Filtered, concentrated to dryness, and lyophilized to obtain about 710 mg of extract.

(Example 2) Measurement of PPARα ligand activity Day 1 of experiment: CV-1 cells (cultured cells derived from male African green monkey kidney) were implanted in a 96-well culture plate so as to be 6 × 10 ³ cells / well, The cells were cultured for about 24 hours under 5% CO ₂ conditions. The medium contains 10% FBS (fetal bovine serum), 10 ml / L penicillin / streptomycin solution (10000 IU / ml, 10000 μg / ml, GIBCO), 37 mg / L ascorbic acid (Wako Pure Chemical Industries, Ltd.) Dulbeccos Modifid Eagle Meduim (GIBCO) was used.

  Experiment Day 2: After washing the cells with OPTI-MEM (GIBCO), pM-PPARα and 4 × UASg-luc were transfected using Lipofectamine Plus (GIBCO). PM-PPARα is a chimeric protein in which a yeast-derived transcription factor GAL4 gene (amino acid sequence 1-147) and a human PPARα ligand binding site gene (amino acid sequence 167-468) are linked to a mammalian expression plasmid pM (Clontech). The 4 × UASg-luc is a reporter plasmid in which a GAL4 response element (UASg) is incorporated four times upstream of the luciferase gene.

  Experiment Day 3: About 24 hours after transfection, the medium was replaced with the medium containing the sample and cultured for 24 hours (n = 4). Samples dissolved in dimethyl sulfoxide (DMSO) were added to the medium at 1/1000 volume to obtain the concentrations shown in Table 1. DMSO was used as an untreated control, and bezafibrate, which is PPARα ligand activity, was used as a positive control.

Experiment Day 4: After washing the cells with Ca, Mg-containing phosphate buffered saline (PBS +), Lucright ^™ (Perkin Elmer), a reagent for measuring luminescence by luciferase, was added, and TopCount ^™ microscintillation / The luminescence intensity of luciferase was measured with a luminescence counter (Perkin Elmer). The ratio to the luciferase luminescence intensity of the untreated control was taken as the luciferase specific activity of the sample.
It measured using pM (plasmid which removed the PPAR (alpha) ligand binding site gene) instead of pM-PPAR (alpha) as a control group similarly to the measurement group. For each sample, the ratio (measurement group / control group) of the average luminescence intensity (n = 4) of the measurement group and the control group was calculated, and the specific activity relative to the untreated control was taken as the PPARα ligand activity of the sample.

実施例１で得られたコンニャク抽出物に、有意なPPARαリガンド活性が認められた。
（実施例３）ＰＰＡＲγリガンド活性測定
実験１日目： ＣＶ−１細胞（雄性アフリカミドリザル腎臓由来の培養細胞）を９６穴培養プレートに６×１０³ｃｅｌｌｓ／ｗｅｌｌとなるように植え込み、３７℃、５％ＣＯ₂条件下で約２４時間培養した。培地には、１０％ＦＢＳ（ウシ胎児血清）、１０ｍｌ／Ｌ ペニシリン・ストレプトマイシン溶液（10000IU/ml、10000μg/ml、GIBCO社）、３７ｍｇ／Ｌ アスコルビン酸（和光純薬工業株式会社）を含むＤＭＥＭ（Dulbeccos Modifid Eagle Meduim：GIBCO社）を用いた。

Significant PPARα ligand activity was observed in the konjac extract obtained in Example 1.
(Example 3) Measurement of PPARγ ligand activity Day 1 of experiment: CV-1 cells (cultured cells derived from male African green monkey kidney) were implanted in a 96-well culture plate so as to be 6 × 10 ³ cells / well, The cells were cultured for about 24 hours under 5% CO ₂ conditions. The medium contains 10% FBS (fetal bovine serum), 10 ml / L penicillin / streptomycin solution (10000 IU / ml, 10000 μg / ml, GIBCO), 37 mg / L ascorbic acid (Wako Pure Chemical Industries, Ltd.) Dulbeccos Modifid Eagle Meduim (GIBCO) was used.

  Experiment Day 2: After washing the cells with OPTI-MEM (GIBCO), pM-mPPARγ and 4 × UASg-luc were transfected using Lipofectamine Plus (GIBCO). PM-mPPARγ is a chimeric protein expression plasmid in which a yeast-derived transcription factor GAL4 gene (amino acid sequence 1-147) and a mouse PPARγ ligand binding site gene (amino acid sequence 174-475) are combined, and 4 × UASg-luc is This is a reporter plasmid in which a GAL4 response element (UASg) is incorporated four times upstream of the luciferase gene.

  Experiment Day 3: About 24 hours after transfection, the medium was replaced with the medium containing the sample and cultured for 24 hours (n = 4). Samples dissolved in dimethyl sulfoxide (DMSO) were added to the medium at 1/1000 volume to obtain the concentrations shown in Table 2. DMSO was used as an untreated control, and troglitazone, which is PPARγ ligand activity, was used as a positive control.

Experiment Day 4: After washing the cells with Ca, Mg-containing phosphate buffered saline (PBS +), Lucright ^™ (Perkin Elmer), a reagent for measuring luminescence by luciferase, was added, and TopCount ^™ microscintillation / The luminescence intensity of luciferase was measured with a luminescence counter (Perkin Elmer). The ratio to the luciferase luminescence intensity of the untreated control was taken as the luciferase specific activity of the sample.
In the same manner as in the measurement group, measurement was performed using pM (a plasmid from which the PPARγ ligand binding site gene was removed) instead of pM-mPPARγ as a control group. For each sample, the ratio (measurement group / control group) of the average value (n = 4) of the luminescence intensity of the measurement group and the control group was calculated, and the specific activity relative to the untreated control was defined as the PPARγ ligand activity of the sample.

実施例１で得られたコンニャク抽出物に、有意なPPARγリガンド活性が認められた。

Significant PPARγ ligand activity was observed in the konjac extract obtained in Example 1.

(Example 4) Manufacture of konjac extract-containing tablets Konjac extract (Example 1) 45 parts by weight lactose 35 parts by weight Crystalline cellulose 15 parts by weight Sucrose fatty acid ester 5 parts by weight Eating and drinking tablets containing the extract were produced.

Example 5 Production of Konjac Extract-Containing Soft Capsules Konjac Extract (Example 1) 40 parts by weight olive oil 55 parts by weight glycerin fatty acid ester 5 parts by weight The above composition is mixed, pulverized, filled into gelatin capsules, and konjac A capsule for food and drink containing the extract was produced.

(Example 6) Production of soft drink containing konjac extract Konjac extract (Example 1) 0.5 parts by weight liquid sugar 30 parts by weight Citric acid 0.1 parts by weight Fragrance 0.5 parts by weight Water 150 parts by weight Above Mixing in composition produced a soft drink containing konjac extract.

(Example 7) Preparation of konjac extract-containing cracker Konjac extract (Example 1) 1 part by weight flour 120 parts by weight salt 1 part by weight baking powder 2 parts by weight butter 30 parts by weight water 40 parts by weight A konjac extract-containing cracker was prepared.

(Example 8) Skonkonyaku extract containing konjac extract (Example 1) 10 parts by weight soft flour 100 parts by weight baking powder 5 parts by weight salt 0.5 parts by weight sugar 15 parts by weight butter 40 parts by weight beaten egg 5 parts by weight sour cream 10 parts by weight milk 10 parts by volume The above composition was mixed to prepare a scone containing a konjac extract.

(Example 9) Preparation of konjac extract-containing dressing Konjac extract (Example 1) 10 parts by weight olive oil 80 parts by weight Vinegar 60 parts by weight Salt 3 parts by weight Pepper 1 part by weight Lemon water 5 parts by weight An extract-containing dressing was prepared.

(Example 10) Preparation of chewing gum containing konjac extract Konjac extract (Example 1) 0.5 parts by weight gum base 20 parts by weight Sugar 70 parts by weight Fragrance 0.5 parts by weight Water A containing chewing gum was prepared.

(Example 11) Preparation of konjac extract-containing tea 0.1 part by weight of konjac extract (Example 1) was suspended in tea.

Claims (16)

Peroxisome proliferator-responsive receptor ligand composition comprising konjac plant ( Amorphophallus ) material as an active ingredient. The peroxisome proliferator-responsive receptor ligand composition according to claim 1, comprising an organic solvent extract of konjac plant ( Amorphophallus ) material as an active ingredient. The peroxisome proliferator-responsive receptor ligand composition according to claim 1 or 2, wherein the organic solvent extract of konjac plant ( Amorphophallus ) material is an extract extracted with ethanol at a concentration of 10% by volume or more. The peroxisome proliferator-responsive receptor ligand composition according to any one of claims 1 to 3, wherein the konjac plant ( Amorphophallus ) is konjac ( Amorphophallus konjac K. Koch ). The peroxisome proliferator-responsive receptor ligand composition according to any one of claims 1 to 4, wherein the konjac plant ( Amorphophallus ) material is either konjac powder or processed konjac.   The peroxisome proliferator response according to any one of claims 1 to 5, wherein the peroxisome proliferator-responsive receptor ligand composition is at least one of the peroxisome proliferator-responsive receptors α, γ, and δ. Sex receptor ligand composition.   A composition for preventing or treating insulin resistance comprising the peroxisome proliferator-responsive receptor ligand composition according to any one of claims 1 to 6.   A composition for preventing or treating lifestyle-related diseases, comprising the peroxisome proliferator-responsive receptor ligand composition according to any one of claims 1 to 6.   The composition for preventing or treating a lifestyle-related disease according to claim 8, wherein the lifestyle-related disease is metabolic syndrome.   The composition for preventing or treating a lifestyle-related disease according to claim 8, wherein the lifestyle-related disease is at least one disease selected from the group consisting of obesity, obesity, diabetes, hyperlipidemia, hypertension, and gout.   A food or beverage comprising the peroxisome proliferator-responsive receptor ligand composition according to any one of claims 1 to 6.   A cosmetic comprising the peroxisome proliferator-responsive receptor ligand composition according to any one of claims 1 to 6.   A pharmaceutical comprising the peroxisome proliferator-responsive receptor ligand composition according to any one of claims 1 to 6.   A method for preventing or treating insulin resistance or lifestyle-related diseases using the peroxisome proliferator-responsive receptor ligand composition according to any one of claims 1 to 6. A method for producing a peroxisome proliferator-responsive receptor ligand composition, comprising extracting a konjac plant ( Amorphophallus ) material using an organic solvent.   The method for producing a peroxisome proliferator-responsive receptor ligand composition according to claim 15, wherein the organic solvent is ethanol having a concentration of 10% by volume or more.