JP2013159579A - Ppar activator - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a peroxisomal proliferator activated receptor activator.SOLUTION: A PPAR activator includes malt as an active ingredient. Alternatively, the PPAR activator includes malt treated with water as an active ingredient.

Description

  The present invention includes fatty acid metabolism activation, body fat burning promotion, adipocyte differentiation promotion, obesity prevention / improvement, insulin resistance prevention / improvement, diabetes prevention / improvement, dyslipidemia prevention / improvement, fatty liver prevention / improvement, arteries Peroxisome proliferator activated receptor (referred to as PPAR) activator effective for prevention and improvement of sclerosis, improvement of endurance, prevention and improvement of cardiac hypertrophy, prevention and improvement of ischemic heart disease, etc. .

  Low-molecular-weight lipophilic ligands such as steroids, thyroid hormones, and retinoids have various physiological functions such as morphogenesis in ontogenesis, cell proliferation, differentiation, and maintenance of homeostasis through ligand-specific nuclear receptors. It is involved in regulation. PPAR is one of the nuclear receptors, and was identified in 1990 as a protein that mediates the action of increasing peroxisomes, which are intracellular organelles involved in lipolysis, meaning that it is activated by peroxisome proliferators Was named Peroxisome Proliferator Activated Receptor α (peroxisome proliferator activated receptor: PPARα). Thereafter, δ type and γ type were identified as isoform genes structurally similar to α type, and it is known to have a total of three subtypes.

Each subtype of PPAR is activated in a ligand-dependent manner, and forms a heterodimer with RXR (Retinoid X Receptor) having 9-cis retinoic acid as a ligand, thereby causing a PPAR responsive element (PPAR responsive element) in the promoter region. ; The expression of various genes having PPRE) is controlled (Non-patent Documents 1 and 2).
For example, a reporter assay using PCO of ACO (Acyl-CoA oxidase), which is known as a key enzyme for fatty acid β-oxidation, has been performed. According to this report, linoleic acid known as a PPAR ligand is each of PPARα, δ and γ. It has been reported that the ACO transcriptional activity is enhanced through the process (Non-patent Document 3).
As will be described below, in recent years it has become clear that PPAR is involved in numerous physiological and pathological phenomena.

  Specifically, the function of PPARα is considered to be widely involved in the maintenance of energy metabolism and homeostasis in the living body such as fatty acid synthesis / transport / secretion and ATP production in fat consuming organs. In particular, gene expression of β-oxidation-related enzymes (ACO, HMG-CoA synthase, Acyl-CoA synthase, Medium chain acyl-CoA dehydrogenase, Fatty acid binding protein, Lipoprotein lipase, etc.) important for fatty acid metabolism is strongly dependent on PPARα activation It has become clear that PPARα is highly expressed in the liver, heart, kidney, etc., and PPARα activators are widely recognized as being effective in activating lipid metabolism in these organs.

  Activation of fatty acid metabolism accompanying the activation of PPARα is thought to lead to degradation of liver fat, improvement of fatty liver, promotion of decomposition and burning of body fat such as visceral fat and subcutaneous fat, and suppression of obesity. Fibrate drugs known as PPARα activators have been shown to have an action of promoting fatty acid combustion, an action of increasing HDL cholesterol, and an action of increasing the expression of adiponectin receptor in recent years. Are widely used as therapeutic agents for hyperglycemia, dyslipidemia, hyperglycemia, atherosclerosis and the like (Non-patent Document 4, Patent Documents 1 and 2). In addition, it has been reported that PPARα activators are effective for cardiac hypertrophy and ischemic heart disease (Non-patent Documents 5 and 6).

  Therefore, PPARα activator activates fatty acid metabolism, promotes fat burning, prevents and improves obesity, prevents and improves dyslipidemia, prevents and improves fatty liver, prevents and improves insulin resistance and diabetes It is considered to be widely effective in the prevention and improvement of arteriosclerosis, the prevention and improvement of cardiac hypertrophy and ischemic heart disease, etc. In recent years, search and development of PPARα activators have been actively conducted (patents) Literature 3-5).

  PPARδ (also known as PPARβ, NUC1, FAAR) has not been clarified for a long time since it was cloned in 1992, but in recent years, it has been studied through the use of genetically modified animals and the development of selective agonists for PPARδ. It has become clear that it has various physiological functions. In studies using mice overexpressing PPARδ, suppression of body weight increase due to high-fat diet load, reduction of fat weight, reduction of blood neutral fat, and suppression of fatty liver were observed (Non-patent Document 7). ). In an experiment in which GW501516, a PPARδ selective agonist, was administered to obese rhesus monkeys, HDL cholesterol was increased and neutral fat and LDL cholesterol were decreased (Non-patent Document 8).

  In addition, GW501516 was allowed to act on skeletal muscle-derived cells, and the effects on cellular gene expression were examined. As a result, fatty acid uptake and transport, expression of fatty acid metabolism-related genes such as mitochondrial β-oxidation enzymes, uncoupling proteins, etc. It has been shown to induce. Furthermore, in mice administered with GW501516, as with adipose tissue-specific PPARδ overexpressing mice, suppression of body weight increase and decrease in fat weight were observed due to high fat diet loading, and it was also revealed that insulin resistance was improved. ing. In this mouse skeletal muscle, fatty acid metabolism-related genes and induction of fatty acid β oxidation have been confirmed. Therefore, activation of PPARδ suppresses fat accumulation in peripheral tissues by increasing energy consumption of skeletal muscle. It is considered that this may improve insulin resistance (Non-patent Document 9). In addition, administration of GW501516 to genetically obese mice suppressed pancreatic islet hypertrophy, and is considered to have a protective effect on pancreatic islets (Non-patent Document 9).

  In addition, it has been reported that obesity and insulin resistance are suppressed by overexpressing PPARδ specifically in skeletal muscle. Surprisingly, the skeletal muscle of this mouse has a very high percentage of endurance muscle fibers, which are generally called red muscle and contain a lot of mitochondria. As a result, this mouse is about 2% of the control mouse. It has been shown that it is a mouse with extremely excellent endurance that can run twice as long (Non-patent Document 10). Therefore, it has been considered that the activation of PPARδ is effective in improving exercise endurance.

  Recently, it has also been reported that PPARδ regulates insulin sensitivity in the liver. As a mechanism for this, activation of PPARδ is caused by glucose from the liver via activation of the glycolytic system of the liver and the pentose phosphate cycle. It has been suggested to decrease the supply of insulin and increase insulin sensitivity (Non-patent Document 11).

Thus, the activation of PPARδ is an increase in HDL cholesterol, a decrease in LDL cholesterol, suppression of obesity, improvement of insulin resistance / insulin sensitivity, activation of fatty acid metabolism, promotion of burning of body fat, PPARδ activator is a prophylactic / ameliorating agent for arteriosclerosis, a prophylactic / improving agent for obesity, a prophylactic / improving agent for insulin resistance. It is considered to be effective as a fatty acid oxidation activator, body fat combustion promoter, dyslipidemia preventive / ameliorating agent, fatty liver preventive / ameliorating agent, and endurance improving agent.
Therefore, search and development of PPARδ activators have been actively conducted for such purposes, and flavones, phenoxyacetic acid derivatives and the like have been reported so far (Patent Documents 6 and 7).

  PPARγ is a molecule that acts on energy storage in the presence of sufficient nutrients and acts as a so-called thrifty gene. In particular, PPARγ2 is expressed with relatively strong specificity in adipocytes, and has been shown to play a central role in adipocyte differentiation. Thiazolidine derivatives, known as PPARγ ligands, are known to downsize adipocytes and increase insulin sensitivity by strongly inducing adipocyte differentiation, and are widely used as insulin resistance improvers and diabetes therapeutics. (Non-Patent Document 12).

  Thus, PPAR activators activate fatty acid metabolism, promote fat burning, prevent / improve obesity, prevent / improve dyslipidemia, prevent / improve fatty liver, improve endurance, insulin resistance It is widely effective for prevention and improvement of diabetes and diabetes, prevention and improvement of arteriosclerosis, prevention and improvement of cardiac hypertrophy and ischemic heart disease, etc. Also for prevention and improvement of so-called lifestyle-related diseases and visceral fat syndrome (metabolic syndrome) It is considered effective.

  Malt is a germination of seeds such as barley and wheat, and barley malt is particularly widely used as a raw material for alcoholic beverages and sugars. Beverage obtained by extracting dried black wheat malt with high-temperature water or water vapor, having a blood pressure lowering action, a renal function activating action, a liver function improving action, an obesity preventing action, an alcohol metabolism promoting action or a deodorizing action (Patent Document 8) Or an extract obtained by removing a low molecular fraction from water or a hydrophilic solvent extract of a malt saccharified filtration residue has an action of inhibiting the synthesis of heat shock proteins belonging to the HSP60 family, and is associated with type I diabetes and chronic joints. It is known that it is effective in the treatment of autoimmune diseases such as rheumatism (Patent Document 9).

  However, it has not been known that malt or a preparation obtained by subjecting it to a specific treatment is involved in PPAR activation and is effective in treating the various diseases described above involving PPAR.

Special Table 2002-502869 Special Table 2002-533410 Publication JP 2001-354558 A JP 2002-80362 A Japanese Patent Laid-Open No. 2003-34636 JP 2007-119429 A Special Table 2007-536343 JP 2002-95443 A Japanese Patent Laid-Open No. 9-216829

Cell: 31 (6) 218-234, 1999 J Lipid Res. 37, 907-925, 1996 J Biol Chem. 274, 23368-23377, 1999 Curr Opin Lipidol. 10, 151-159, 1999 Endcrinology 144, 4187-4194, 2003 Circulation 108, 2393-2399, 2003 Cell. 113, 159-170, 2003 Proc. Natl. Acad. Sci. USA. 98, 5306-5311, 2001 Proc. Natl. Acad. Sci. USA. 100, 15924-15929, 2003 PloS Biol. 2, e294, 2004 Proc. Natl. Acad. Sci. USA. 103, 3444-3449, 2006 Diabetes. 49, 759-767, 2000

  The present invention relates to providing a food, drug, or quasi-drug having an excellent PPAR activation action and high safety.

  The present inventors have selected PPARα, a malt, a water-treated product of malt, or a filtered residue of a water-treated product of malt, or an organic solvent extract obtained therefrom from among natural product materials rich in food experience. There is an activation effect on PPARδ and PPARγ, activation of fatty acid metabolism, promotion of fat burning, promotion of adipocyte differentiation, prevention and improvement of obesity, prevention and improvement of insulin resistance, prevention and improvement of diabetes, prevention and improvement of dyslipidemia, Formulated as an active ingredient in foods, pharmaceuticals or quasi drugs that demonstrate effects such as fatty liver prevention / improvement, arteriosclerosis prevention / improvement, endurance improvement, prevention / improvement of cardiac hypertrophy, prevention / improvement of ischemic heart disease, etc. It was found to be useful as a material to be used.

That is, this invention provides the PPAR activator which uses malt as an active ingredient.
In one embodiment, the PPAR activator comprises a malt water treatment product as an active ingredient.
In another embodiment, the PPAR activator comprises a filtration residue of a malt water treatment product as an active ingredient.
In another embodiment, the PPAR activator comprises, as an active ingredient, an organic solvent extract obtained from the filtration residue of the malt water treatment product or the malt.
Moreover, this invention provides the fatty-acid-metabolizing activator which uses the organic-solvent extract obtained from the filtration residue of the water treatment material of the said malt, or the said malt as an active ingredient.
Moreover, this invention provides the body fat combustion promoter which uses the organic solvent extract obtained from the filtration residue of the water treatment thing of the said malt, or the said malt as an active ingredient.
Moreover, this invention provides the adipocyte differentiation promoter which uses the organic solvent extract obtained from the filtration residue of the water treatment thing of the said malt, or the said malt as an active ingredient.
Moreover, this invention provides the obesity prevention and improvement agent which uses the organic-solvent extract obtained from the filtration residue of the water treatment thing of the said malt, or the said malt as an active ingredient.
Moreover, this invention provides the insulin resistance preventive / improving agent which uses the organic-solvent extract obtained from the filtration residue of the water treatment thing of the said malt, or the said malt as an active ingredient.
Moreover, this invention provides the diabetes preventive / improving agent which uses the organic-solvent extract obtained from the filtration residue of the water treatment thing of the said malt, or the said malt as an active ingredient.
The present invention also provides an agent for preventing or improving dyslipidemia, comprising as an active ingredient an organic solvent extract obtained from the filtration residue of the water-treated product of malt or the malt.
The present invention also provides a fatty liver preventive / improving agent comprising, as an active ingredient, an organic solvent extract obtained from the filtration residue of the malt water-treated product or from the malt.
The present invention also provides an arteriosclerosis preventive / ameliorating agent comprising as an active ingredient an organic solvent extract obtained from the filtration residue of the water-treated product of malt or from the malt.
Moreover, this invention provides the endurance improver which uses the organic-solvent extract obtained from the filtration residue of the water treatment thing of the said malt, or the said malt as an active ingredient.
The present invention also provides a preventive / improving agent for cardiac hypertrophy comprising as an active ingredient an organic solvent extract obtained from the filtration residue of the malt water-treated product or from the malt.
The present invention also provides a preventive / ameliorating agent for ischemic heart disease comprising as an active ingredient an organic solvent extract obtained from the filtration residue of the malt water-treated product or from the malt.
The present invention further provides a method for producing a PPAR activator by preparing an organic solvent extract from malt.
In one embodiment, in the production method, an organic solvent extract is prepared from a water-treated product of malt.
In another embodiment, in the production method, malt is filtered after water treatment, and an organic solvent extract is prepared from the obtained filtration residue.

  Since the PPAR activator of the present invention has an excellent PPAR activation action and is highly safe even if ingested for a long period of time, activation of fatty acid metabolism, promotion of fat burning, promotion of adipocyte differentiation, prevention of obesity・ Improvement, prevention and improvement of insulin resistance, prevention and improvement of diabetes, prevention and improvement of dyslipidemia, prevention and improvement of fatty liver, prevention and improvement of arteriosclerosis, improvement of endurance, prevention and improvement of cardiac hypertrophy, prevention of ischemic heart disease It is useful as a material to be blended as an active ingredient in foods and beverages, pharmaceuticals or quasi drugs that exhibit effects such as prevention and improvement.

The graph which showed PPAR (alpha) activation ability. A: Control (ethanol), B: 0.01% lyophilized filtrate of malt water treatment product, C: 0.01% malt organic solvent extract, D1 to D3: Filtration residue of malt water treatment product Organic solvent extract, 0.002%, 0.005% and 0.01%, respectively, Wy: Wy14643 10 μM. Error bars = ± SE (n = 3), * indicates a significant difference P <0.05 with respect to Control. The graph which showed PPAR (delta) activation ability. A: Control (ethanol), B: 0.01% lyophilized filtrate of malt water treatment product, C: 0.01% malt organic solvent extract, D1 to D3: Filtration residue of malt water treatment product Organic solvent extract, 0.002%, 0.005% and 0.01% respectively, GW: GW501516 1 nM. Error bars = ± SE (n = 3), * indicates a significant difference P <0.05 with respect to Control. The graph which showed PPAR (gamma) activation ability. A: Control (ethanol), B: 0.01% lyophilized filtrate of malt water treatment product, C: 0.01% malt organic solvent extract, D1 to D3: Filtration residue of malt water treatment product Organic solvent extract, 0.002%, 0.005% and 0.01%, respectively, Pio: Pioglitazone 10 μM. Error bars = ± SE (n = 3), * indicates a significant difference P <0.05 with respect to Control.

  As an active ingredient such as a PPAR activator of the present invention, malt, a water-treated product of malt, a filtration residue of a water-treated product of malt, or an organic solvent extract thereof can be used.

  In the present invention, malt refers to germinating seeds of gramineous cereals such as barley, wheat, rye, hen, hadaka and corn. Of these, barley malt can be suitably used. The malt is preferably germinated in a state where the husks are attached. The germination conditions are not particularly limited, but those in which the germinated leaf buds have not yet extended from the shell are preferable. Moreover, it is preferable to remove roots.

  The malt may be used in the present invention as it is, but may be used after being dried, roasted, pulverized, dried or dried and pulverized in advance. As drying conditions, drying at 32 to 65 ° C. until the water content in the malt is 10% by mass or less, preferably 5% by mass or less is preferable for improving the storage stability of the malt. It is preferable to carry out a drying treatment after drying in order to further improve the storage stability. The roasting temperature is preferably a low temperature from the viewpoint of inhibiting the decomposition of the active ingredient and inhibiting the deactivation of the enzyme in the malt, preferably 80 to 110 ° C, more preferably 80 to 90 ° C. . Preferably, the malt is prepared as a water treatment product of malt, more preferably as a filtration residue of the water treatment product of malt.

  In order to prepare a water-treated product of malt, for example, malt or a dried, roasted or pulverized product thereof may be mixed with water. In the case of the water treatment, starch or saccharide may be further added to the mixed solution of malt and water as necessary. During water treatment, the malt is preferably completely immersed in water. The mass ratio of malt and water is dry malt 1: water 2-10, preferably dry malt 1: water 3-9. Moreover, 40-75 degreeC is preferable during the water treatment, and, as for the temperature of the said malt liquid mixture, More preferably, it is 40-50 degreeC. The treatment time is 1 to 6 hours, preferably 2 to 4 hours. By the water treatment, the malt swells, and it becomes easy to separate the husk, seed coat, germ and the like from those obtained by degrading the endosperm with the malt-derived enzyme.

  The water treatment product of malt is obtained by the above procedure. Or you may utilize the malt saccharified liquid prepared by a normal procedure in manufacturing processes, such as beer, as a water treatment thing of malt. It is preferable in that the active ingredient can be concentrated to filter the obtained water-treated product of malt to remove the liquid component and obtain a filtration residue of the water-treated product including husk, seed coat and germ. The filtration should just be a filtration which can isolate | separate the sediment and floating matter which can be visually recognized with the naked eye. The residue may contain a liquid component derived from a water-treated product of malt.

  More preferably, the malt, the malt water-treated product, or the filtered residue of the malt water-treated product thus obtained is extracted with an organic solvent at room temperature or under heating. The active ingredient can be further concentrated by extracting with an organic solvent. As the organic solvent for extraction, either a polar organic solvent or a nonpolar organic solvent can be used. Examples of the organic solvent include monovalent, divalent or polyhydric alcohols; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; chain or cyclic ethers such as tetrahydrofuran and diethyl ether; Polyethers such as polyethylene glycol; saturated or unsaturated hydrocarbons; aromatic hydrocarbons such as benzene and toluene; halogenated hydrocarbons such as dichloromethane, chloroform, dichloroethane, and carbon tetrachloride; pyridines; dimethyl sulfoxide Acetonitrile; carbon dioxide, supercritical carbon dioxide; fats and oils, waxes, other oils, and the like. Among these, alcohols and saturated hydrocarbons are preferable.

  Although it does not specifically limit as said alcohol, For example, monohydric alcohols, such as methanol, ethanol, propanol, butanol, amyl alcohol, hexanol, heptanol, octanol; 1, 3- butylene glycol, ethylene glycol, propylene glycol, 1 Dihydric alcohols such as 1,4-butanediol, 1,5-pentanediol, 1,6-hexanediol; trihydric or higher alcohols such as glycerin, etc. Among these, monohydric alcohols and divalent alcohols Alcohols are preferable, and monohydric alcohols are more preferable.

  As said alcohol, C1-C10 and C1-C4 are more preferable. Specific examples include methanol, ethanol, 1,3-butylene glycol, n-propanol, isopropanol, n-butanol, isobutanol, sec-butanol, t-butanol and the like, with ethanol and 1,3-butylene glycol being preferred. .

  Examples of the saturated hydrocarbons include linear, branched or cyclic saturated hydrocarbons such as methane, ethane, propane, n-butane, n-pentane, n-hexane, n-heptane, n -Linear saturated hydrocarbons such as octane, n-nonane, n-decane; 2-methylbutane, 2,2-dimethylpropane, 2-methylpentane, 3-methylpentane, 2,2-dimethylbutane, 2,3- Branched chain saturated hydrocarbons such as dimethylbutane, 2-methylhexane, 3-methylhexane, 2,2,4-trimethylpentane; cyclic saturated hydrocarbons such as cyclopentane, cyclohexane, cycloheptane, and the like. Among these, Straight chain saturated hydrocarbons are preferred.

  As said saturated hydrocarbon, C1-C10, C5-C10, and C5-C8 are more preferable. Specifically, n-pentane, n-hexane, n-heptane, n-octane, 2-methylbutane, 2,2-dimethylpropane, 2-methylpentane, 3-methylpentane, 2,2-dimethylbutane, 2 , 3-dimethylbutane, 2-methylhexane, 3-methylhexane, cyclohexane and the like, among which n-hexane is preferable.

  The above organic solvents can be used alone or in admixture of two or more.

  The organic solvent used in the present invention may contain water. As the organic solvent used for the water-containing organic solvent, a hydrophilic organic solvent is preferable. Here, the hydrophilic organic solvent is not particularly limited, but the above-mentioned protic hydrophilic organic solvents such as alcohols, acetic acid and pyridines, and aprotic hydrophilic organic solvents such as ketones, acetonitrile and dimethyl sulfoxide. Of these, a protic hydrophilic organic solvent is preferred. Among these, the above alcohols, more monovalent and divalent alcohols, more alcohols having 1 to 4 carbon atoms, more preferably ethanol and 1,3-butylene glycol are preferable. These solvents can be used alone or in combination as a mixture.

  The water content in the water-containing organic solvent is not particularly limited. For example, when the organic solvent is a monohydric or dihydric alcohol, the water content is 30% by volume or less, preferably 10% by volume or less, more preferably It can be 5% by volume or less. For example, the concentration of the hydrophilic organic solvent in the water-containing organic solvent is at least 70% by volume or more of the hydrophilic organic solvent, more preferably 70 to 100% by volume, still more preferably 90 to 100% by volume, and still more preferably 95%. It can be ˜99.9% by volume.

  The extraction means in the present invention is not particularly limited. For example, liquid-liquid extraction, solid-liquid extraction, immersion, leaching, decoction, reflux extraction, ultrasonic extraction, microwave extraction, centrifugal extraction, supercritical extraction, etc. These may be used alone or in combination of two or more. At this time, a batch type extractor or a Soxhlet extractor may be used. The extraction may be performed in a non-oxidative atmosphere while removing dissolved oxygen by bubbling degassing or inert gas such as nitrogen gas.

  The extraction conditions are not particularly limited, but the extraction temperature may be 0 to 100 ° C., preferably 40 to 75 ° C., more preferably 40 to 50 ° C., and the extraction period may be 10 minutes to 12 hours, preferably 30 minutes to It may be 8 hours, more preferably 1 to 5 hours. Moreover, the usage-amount of the said organic solvent should just be 1-100 mass parts with respect to the filtration residue of the said malt water processed material, Preferably it is 1-50 mass parts, More preferably, it may be 3-40 mass parts.

  As an example of the extraction procedure, an aqueous solution of monovalent or divalent alcohol having a concentration of 90 to 95% by volume (preferably an aqueous ethanol solution) is used as an extraction solvent with respect to 1 part by mass of the filtration residue (in terms of dry matter). Extraction for 30 minutes to 8 hours at 40 to 50 ° C. using part by mass is exemplified.

  After the extraction process, liquid-liquid distribution with water may be performed in order to further remove impurities and the like. Specifically, the solvent is distilled off from the organic solvent extract under reduced pressure, water is added to the remaining oil, and after performing physical means such as mixing, stirring, shaking, and centrifugation, the oil layer is recovered. . You may repeat this operation 1-3 times suitably. The recovered oil layer can be concentrated and used as an active ingredient of the present invention.

  Although the usage-amount of the water in the said liquid-liquid distribution is not specifically limited, It is preferable that it is 2-100 mL with respect to 1g of dried solids. Moreover, it is preferable that extraction temperature is 4-80 degreeC, and it is more preferable that it is 10-40 degreeC, Furthermore, it is 10-30 degreeC.

  The obtained organic solvent extract of the filtration residue of the processed malt water product can be used as it is, but further diluted, concentrated or freeze-dried, and / or prepared in liquid, powder or paste form and used. You can also.

  As shown in Examples below, organic solvent extracts prepared by extracting malt, malt water-treated products or malt water-treated filtration residues with organic solvents are dependent on PPARα, PPARδ, and PPARγ. Has the effect of enhancing the transcriptional activity of various genes. As described above, PPARα, PPARδ, and PPARγ are widely involved in the maintenance of energy metabolism and homeostasis in the living body, and particularly β-oxidation-related enzymes (ACO, HMG-CoA synthase, Acyl-CoA synthase, important for lipid metabolism) The gene expression of medium chain acyl-CoA dehydrogenase, Fatty acid binding protein, Lipoprotein lipase, etc.) strongly depends on the activation of PPARα and / or PPARδ (Non-patent Documents 1-11, Patent Documents 1-7), Since it is known that insulin resistance and diabetes can be improved by induction of adipocyte differentiation by PPARγ ligand (Non-patent Document 12), filtration residue of malt, malt water treatment product or malt water treatment product, or these Organic solvent extract of fatty acid metabolism activation, body fat burning promotion, adipocyte differentiation promotion, obesity prevention / improvement, insulin resistance prevention / improvement, diabetes (I Type and / or type II) prevention / improvement, dyslipidemia prevention / improvement, fatty liver prevention / improvement, arteriosclerosis prevention / improvement, endurance, heart hypertrophy prevention / improvement, ischemic heart disease prevention / improvement, etc. Widely effective.

Accordingly, malt, a water-treated product of malt or a water-treated product of malt, or an organic solvent extract thereof (hereinafter, the preparation of the present invention) has an activating action on PPARα, PPARδ, and PPARγ. , PPARα, PPARδ, and PPARγ activator, fatty acid metabolism activator, body fat burning promoter, adipocyte differentiation promoter, obesity prevention / amelioration agent, insulin resistance prevention / amelioration agent, diabetes (Type I and / or Type II) preventive / ameliorating agent, dyslipidemia preventing / ameliorating agent, fatty liver prevention / improving agent, arteriosclerosis preventing / improving agent, endurance improving agent, cardiac hypertrophy preventing / ameliorating agent, imaginary It can be used as a prophylactic / ameliorating agent for blood heart disease, etc. (hereinafter referred to as “PPAR activator etc.”), and can be used for the production of PPAR activator etc.
In addition, the preparation of the present invention comprises PPARα, PPARδ or PPARγ activation, fatty acid metabolism activation, body fat burning promotion, fat cell differentiation promotion, obesity prevention / improvement, insulin resistance prevention / improvement, diabetes (type I and / or Or type II) Prevention / improvement, dyslipidemia prevention / improvement, fatty liver prevention / improvement, arteriosclerosis prevention / improvement, endurance, heart hypertrophy prevention / improvement, ischemic heart disease prevention / improvement, etc. It can be used as an active ingredient of food or drink for humans or animals, pharmaceuticals, quasi-drugs or cosmetics.
In addition, the preparation of the present invention comprises PPARα, PPARδ or PPARγ activation, fatty acid metabolism activation, body fat burning promotion, fat cell differentiation promotion, obesity prevention / improvement, insulin resistance prevention / improvement, diabetes (type I and / or Or type II) prevention / improvement, dyslipidemia prevention / improvement, fatty liver prevention / improvement, arteriosclerosis prevention / improvement, endurance, heart hypertrophy prevention / improvement, ischemic heart disease prevention / improvement, etc. It can be applied to foods and drinks that are intended and displayed as necessary, such as functional foods, foods for the sick, and foods for specified health use.

Furthermore, according to the present invention, PPAR activity is obtained by preparing an organic solvent extract from malt or a water-treated product of malt, or filtering malt after water treatment and preparing an organic solvent extract from the obtained filtration residue. A method of producing the agent is provided. Preferably, the PPAR activator is an activator for any one or more of PPARα, PPARδ, and PPARγ.
Furthermore, according to the present invention, the organic solvent extract is prepared from malt or a water-treated product of malt, or the malt is filtered after water treatment, and the organic solvent extract is prepared from the obtained filtration residue. Fatty acid metabolism activator, body fat burning promoter, adipocyte differentiation promoter, obesity prevention / amelioration agent, insulin resistance prevention / amelioration agent, diabetes (type I and / or type II) prevention / amelioration agent, dyslipidemia Methods for producing a prophylactic / ameliorating agent, a fatty liver prophylactic / improving agent, an arteriosclerosis prophylactic / improving agent, an endurance enhancing agent, a cardiac hypertrophy prophylactic / improving agent, an ischemic heart disease prophylactic / ameliorating agent, and the like are provided.
The conditions such as malt water treatment, filtration, and solvent extraction are as described above.

Examples of administration forms of pharmaceuticals and quasi drugs using the preparation of the present invention as an active ingredient include oral administration by tablets, capsules, granules, powders, syrups, etc. or injections, suppositories, inhalants And parenteral administration by transdermal absorption agents, external preparations and the like.
In order to prepare such various dosage forms, the preparation of the present invention alone or other pharmaceutically acceptable excipient, binder, extender, disintegrant, interface, Activators, lubricants, dispersants, buffers, preservatives, flavoring agents, fragrances, coating agents, carriers, diluents, and the like can be used in appropriate combinations.
Of these dosage forms, the preferred form is oral administration. For example, when preparing an oral liquid preparation, it can be produced by a conventional method with the addition of a flavoring agent, a buffering agent, a stabilizer and the like. .

  Cosmetics using the preparation of the present invention as an active ingredient can be used as a skin external preparation, a cleaning agent, a bath agent, a makeup cosmetic, or the like. Various agents such as water, massage agents, lotions, emulsions, gels, creams, ointments, powders, packs, poultices, granules, foundations, lipsticks, shampoos, conditioners, hair tonics, tablets, capsules, sheet products Can be provided in molds. In order to prepare cosmetics of such various dosage forms, the above-mentioned preparation of the present invention alone or incorporated in cosmetics, an external base material, oil or oily substance, humectant, powder, pigment, Emulsifiers, solubilizers, cleaning agents, UV absorbers, thickeners, medicinal ingredients, fragrances, resins, antibacterial and antifungal agents, alcohols, chelates, inorganic acids, organic acids, vitamins, water-soluble polymers, A surfactant or the like can be used in appropriate combination.

Examples of food and drink products using the preparation of the present invention as an active ingredient include various types of breads, cakes, noodles, confectionery, jelly, frozen foods, ice creams, dairy products, beverages, soups, etc. In addition to food, examples include the same forms (tablets, capsules, syrups, etc.), beverages, edible oils, and the like as the aforementioned oral preparations.
In addition, in order to prepare such various forms of food and drink, the preparation of the present invention alone or other food and drink materials, solvent, softener, oil, emulsifier, preservative, fragrance, What is necessary is just to prepare by the manufacturing method of well-known food-drinks combining a stabilizer, a coloring agent, antioxidant, a moisturizer, a thickener, etc. suitably. The obtained foods and drinks are foods and drinks for activating PPARα, PPARδ or PPARγ, foods and drinks for activating fatty acid metabolism, foods and drinks for promoting fat burning, foods and drinks for promoting fat cell differentiation, food and drink for preventing and improving obesity Insulin resistance prevention / improvement food / drink, diabetes prevention / improvement food / drink, dyslipidemia prevention / improvement food / drink, fatty liver prevention / improvement food / drink, arteriosclerosis prevention / improvement food / drink, endurance improvement Food / beverage products, food / beverage products for prevention / improvement of cardiac hypertrophy, food / beverage products for prevention / improvement of ischemic heart disease, pet foods, and the like.

Although the compounding quantity of the said preparation of the said invention with respect to the said food-drinks changes with the usage forms, it is 0.0001-10 mass% normally in conversion of the dry matter of the said extract in the whole composition, Furthermore, 0.001-5 mass %, And further preferably 0.002 to 2% by mass.
In the case of pharmaceuticals other than the above, for example, oral solid preparations such as tablets, granules, capsules, and oral liquid preparations such as oral liquids, syrups, etc., the dry matter conversion of the preparation of the present invention in the whole composition In general, the content is preferably 0.01 to 95% by mass, more preferably 5 to 90% by mass, and still more preferably 10 to 50% by mass.

  The dose (effective intake amount) of the foods, beverages, pharmaceuticals, quasi drugs, cosmetics and the like is preferably 1 to 5000 mg / 60 kg body weight per day in terms of dry matter of the preparation of the present invention. More preferably, it is 5-3000 mg / 60 kg body weight, More preferably, it is 10-2000 mg / 60 kg body weight, and it is still more preferable to set it as 100-1000 mg / 60 kg body weight.

  By administering or ingesting the preparation of the present invention, PPARα, PPARδ, and / or PPARγ are activated, fatty acid metabolism activation, body fat burning promotion, adipocyte differentiation promotion, obesity prevention / improvement, insulin resistance prevention / Promote improvement, prevention and improvement of diabetes, prevention and improvement of dyslipidemia, prevention and improvement of fatty liver, prevention and improvement of arteriosclerosis, improvement of endurance, prevention and improvement of cardiac hypertrophy, prevention and improvement of ischemic heart disease, etc. it can. Therefore, the preparation of the present invention can be used in the process for that purpose. The subject of administration or ingestion is particularly limited as long as it is a human or animal in need of PPAR activation or improvement of the related state or function (for example, prevention or improvement of the above-mentioned disease or condition, or improvement of the above-mentioned function). For example, patients with obesity, dyslipidemia, fatty liver, insulin resistance, diabetes (type I and / or type II), arteriosclerosis, cardiac hypertrophy or ischemic heart disease and their reserves, endurance There are those who have weakened and who want to improve.

(Analysis method)
(I) Protein content of the extract The protein content of the extract was calculated from the nitrogen content of the sample measured by the Kjeldahl method using a nitrogen / protein conversion factor of 6.25.
(Ii) Lipid content of the extract The lipid content of the extract was determined by measuring the weight of the lipid extracted from the sample by the Soxhlet extraction method using diethyl ether as the extraction solvent.
(Iii) Ash content of the extract The ash content of the extract was measured by the direct ashing method. Specifically, the sample was incinerated at 550 ° C., and the difference in sample weight before and after incineration was measured.
(Iv) Water content of the extract The water content of the extract was measured by a heat drying method under reduced pressure. Specifically, the sample was dried by heating under reduced pressure at 70 ° C. for 5 hours, and the difference in sample weight before and after drying was measured.

(Production Example 1) Preparation of Organic Solvent Extract of Filtration Residue of Water Treatment Product of Malt After drying germinated barley, malt for beer (Pilsner malt, 800 g) prepared by low temperature roasting was prepared at 40-50 ° C. Was added to 5.2 kg of warm water, and a water-treated product was prepared by reacting for 2-3 hours while maintaining the temperature. 4.4 kg of 95% ethanol was added to the residue obtained by removing the liquid component (wort) by filtration from this water-treated product, and extracted at 40 to 50 ° C. for 2 to 3 hours. The obtained extract was filtered to remove the residue, and ethanol was distilled off from the filtrate under reduced pressure to obtain an oil layer (yield: about 19 g). The obtained oil layer part was used as an organic solvent extract of the filtration residue of the malt water treatment product in ethanol at 0.2% (D1), 0.5% (D2) or 1% (D3) (w / v). And dissolved as a test substance in the following Examples.
In addition, the organic solvent extract of the filtration residue of the water treatment product of malt is protein 2.4% (w / w), lipid 40.7% (w / w), ash content 1.1% (w / w), The water content was 41.9% (w / w).

(Production Example 2) Preparation of organic solvent extract of malt After drying germinated barley, 150 g of 95% ethanol and 150 g of water were added to malt for beer (Pilsner malt, 50 g) prepared by low-temperature drying. Extracted for 3 hours at ° C. The obtained extract was filtered to remove the residue, and ethanol was distilled off from the filtrate under reduced pressure to obtain an organic solvent extract of malt (yield about 3 g). The obtained oil layer part was dissolved in ethanol so as to be 1% (w / v) as an organic solvent extract of malt, and used as test substance C in the following Examples.

(Production Example 3) Preparation of filtrate lyophilized product of malt water treatment product The filtrate produced during filtration of the water treatment product of malt in Production Example 1 was lyophilized and 1% (w / v) in 50% ethanol. It dissolved so that it might become, and it used as the test substance B of the following Example.

Example 1: PPARα activation test Using human colon total RNA (Clontech), PCR was performed to amplify a PPARα ligand binding site (NCBI RefSeq NM_001001928, nt183-1586, SEQ ID NO: 1). The PCR amplification product was cloned into pCR Blunt (Invitrogen), and a DNA fragment was prepared by restriction enzyme (MluI, KpnI; Takara) treatment. The prepared DNA fragment was inserted into the multicloning site (MluI / KpnI) of pBIND vector (Promega) to obtain pBIND-PPARα LBD. When this vector is introduced into a cell, it expresses a fusion protein of a PPARα ligand binding site and a GAL4 DNA binding site, and the fusion protein binds to a PPARα ligand to bind to a GAL4 binding sequence and activates transcription of a downstream gene. It is to become. In addition, a Renilla luciferase gene is separately incorporated into this vector, and the efficiency of vector introduction can be determined.
African green monkey kidney cell line CV-1 was plated on a 24-well plate and cultured in DMEM (5% charcoal-treated fetal bovine serum) for 1 day. Using a transfection reagent (Superfect transfection reagent; QIAGEN) so that the reporter plasmid (pG5-Luc; Promega) containing the GAL4 binding sequence upstream of the firefly luciferase gene and pBIND-PPARα LBD simultaneously each become 0.2 μg / well. Introduced. Thereafter, the culture solution was replaced with a DMEM (-FBS) medium containing 1/100 amount of the test substance prepared in Production Examples 1 to 3, and further cultured for 1 day. The same amount of ethanol was used as a control. As a positive control, 10 μM Wy14643 (obtained from BIOMOL (Plymouth Meeting DA)) was used.
After washing with PBS, cells were lysed using a dual luciferase assay system (Promega), a substrate solution containing luciferin was added to the lysate, and firefly and Renilla luciferase activities were measured using a luminometer.
In this experimental system, a PPARα activator was searched by measuring the transcriptional activity of a PPARα-dependent gene (luciferase activity: Luc activity). In addition, the transcriptional activity (Luc activity) of a PPARα-dependent gene was defined as follows.

PPARα-dependent gene transcriptional activity (Luc activity)
= (Firefly Luc activity by pG5-Luc) / (Renilla Luc activity by pBIND-PPARα LBD)

  The PPARα activation ability of each test substance is shown in FIG. From FIG. 1, the organic solvent extract of malt or the organic solvent extract of the filtration residue of the water-treated product of malt (Production Examples 1 and 2, C and D1 to D3 in FIG. 1) has PPARα activation ability. I understand.

Example 2: PPARδ activation test A PPARδ activation test was conducted in the same manner as in Example 1.
PCR was performed using Rat IEC-6 total RNA to amplify the PPARδ ligand binding site (NCBI RefSeq NM — 011145, nt690-1595, SEQ ID NO: 2). The PCR amplification product was cloned into pCR Blunt (Invitrogen), and a DNA fragment was prepared by restriction enzyme (MluI, KpnI; Takara) treatment. The prepared DNA fragment was inserted into the multicloning site (MluI / KpnI) of pBIND vector (Promega) to obtain pBIND-PPARδ LBD.
Cells introduced with the vector in the same procedure as in Example 1 were cultured with a test substance, luciferase activity was measured, and a PPARδ activator was searched. As a positive control, GW501516 (ALEXIS BIOCHEMICALS) 1 nM was used. PPARδ-dependent gene transcriptional activity (Luc activity) was defined as follows.

PPARδ-dependent gene transcriptional activity (Luc activity)
= (Firefly Luc activity by pG5-Luc) / (Renilla Luc activity by pBIND-PPARδ LBD)

  FIG. 2 shows the PPARδ activation ability of each test substance. From FIG. 2, the organic solvent extract of the malt or the organic solvent extract of the filtration residue of the water-treated product of malt (Production Examples 1 and 2, C and D1 to D3 in FIG. 2) has PPARδ activation ability. I understand.

Example 3 PPARγ Activation Test A PPARγ activation test was conducted in the same manner as in Example 1.
PCR was performed using Human colon total RNA (Clontech) to amplify the PPARγ2 ligand binding site (NCBI RefSeq NM — 015869, nt703-1606, SEQ ID NO: 3). The PCR amplification product was cloned into pCR Blunt (Invitrogen), and a DNA fragment was prepared by restriction enzyme (MluI, KpnI; Takara) treatment. The prepared DNA fragment was inserted into the multicloning site (MluI / KpnI) of pBIND vector (Promega) to obtain pBIND-PPARγ LBD.
Cells introduced with the vector in the same procedure as in Example 1 were cultured with a test substance, luciferase activity was measured, and a PPARγ activator was searched. Pioglitazone (CAYMAN) 10 μM was used as a positive control. PPARγ-dependent gene transcriptional activity (Luc activity) was defined as follows.

PPARγ-dependent gene transcriptional activity (Luc activity)
= (Firefly Luc activity by pG5-Luc) / (Renilla Luc activity by pBIND-PPARγ LBD)

  FIG. 3 shows the PPARγ activation ability of each test substance. 3. From FIG. 3, the organic solvent extract of malt or the organic solvent extract of the filtration residue of the water-treated product of malt (Production Examples 1 and 2, C and D1 to D3 in FIG. 3) has PPARγ activation ability. I understand.

As mentioned above, it turns out that the organic-solvent extract obtained from the filtration residue of the malt or the water-treated product of malt has the activation effect | action with respect to PPAR (alpha), PPAR (delta), and PPAR (gamma). That is, it can be used as an activator for any one or more of PPARα, PPARδ, and PPARγ.
Furthermore, since the organic solvent extract obtained from the filtration residue of malt or malt water treatment product has PPARα, PPARδ, or PPARγ activation action, as described above, activation of fatty acid metabolism, burning of body fat Promotion, promotion of adipocyte differentiation, prevention and improvement of dyslipidemia, prevention and improvement of obesity, prevention and improvement of fatty liver, improvement of endurance, prevention and improvement of insulin resistance and diabetes (type I and / or type II) It is considered effective for prevention / improvement of arteriosclerosis, prevention / improvement of cardiac hypertrophy, prevention / improvement of ischemic heart disease, and the like.

Claims (23)

  A PPAR activator comprising malt as an active ingredient.   The PPAR activator according to claim 1, comprising a water-treated product of malt as an active ingredient.   The PPAR activator according to claim 1 or 2, wherein the active ingredient is a filtration residue of water-treated malt.   The PPAR activator according to any one of claims 1 to 3, wherein the active ingredient is an organic solvent extract obtained from a filtration residue of the water-treated product of malt or from the malt.   The PPAR activator according to any one of claims 1 to 4, wherein the PPAR is any one or more of PPARα, PPARδ, and PPARγ.   The PPAR activator according to any one of claims 2 to 5, wherein the water-treated product of malt is obtained by immersing malt in water at 40 to 75 ° C.   The PPAR activator according to any one of claims 2 to 6, wherein the water-treated product of malt is obtained by immersing malt in water for 1 to 6 hours.   A fatty acid metabolism activator comprising, as an active ingredient, an organic solvent extract obtained from a filtration residue of the water-treated product of malt or from the malt.   A body fat combustion accelerator comprising, as an active ingredient, an organic solvent extract obtained from a filtration residue of the water-treated product of malt or from the malt.   An adipocyte differentiation promoter comprising as an active ingredient an organic solvent extract obtained from a filtration residue of the water-treated product of malt or from the malt.   An obesity preventing / ameliorating agent comprising, as an active ingredient, an organic solvent extract obtained from a filtered residue of the malt water-treated product or from the malt.   An insulin resistance preventive / improving agent comprising, as an active ingredient, an organic solvent extract obtained from a filtration residue of the malt water-treated product or from the malt.   A diabetes preventive / ameliorating agent comprising as an active ingredient an organic solvent extract obtained from a filtration residue of the water-treated product of malt or from the malt.   An agent for preventing or improving dyslipidemia comprising an organic solvent extract obtained from a filtration residue of a water-treated product of malt or from the malt as an active ingredient.   A fatty liver preventive / improving agent comprising as an active ingredient an organic solvent extract obtained from a filtration residue of the water-treated product of malt or from the malt.   An arteriosclerosis preventive / improving agent comprising, as an active ingredient, an organic solvent extract obtained from a filtered residue of the malt water-treated product or from the malt.   The endurance improver which uses the organic solvent extract obtained from the filtration residue of the water treatment thing of the said malt, or the said malt as an active ingredient.   A preventive / improving agent for cardiac hypertrophy comprising, as an active ingredient, an organic solvent extract obtained from the filtration residue of the water-treated product of malt or from the malt.   A prophylactic / ameliorating agent for ischemic heart disease comprising, as an active ingredient, an organic solvent extract obtained from a filtered residue of the malt water-treated product or from the malt.   A method for producing a PPAR activator by preparing an organic solvent extract from malt.   The method according to claim 20, wherein the malt is filtered after water treatment, and an organic solvent extract is prepared from the obtained filtration residue.   The method according to claim 21, wherein the water treatment is a treatment of immersing the malt in water at 40 to 75 ° C.   The method according to claim 21 or 22, wherein the water treatment is a treatment of immersing the malt in water for 1 to 6 hours.

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