JP2011037811A - Fat-decreasing composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide the new uses of Dioscorea esculenta and/or Angelica keiskei. <P>SOLUTION: A body fat-decreasing composition is characterized by including a substance originated from Dioscorea esculenta and/or a substance originated from Angelica keiskei as active ingredients. A treated product of the Dioscorea esculenta is used as the substance originated from the Dioscorea esculenta. A chalcone-containing substance is used as the substance originated from the Angelica keiskei. The body fat-decreasing composition exhibits an action for improving the expression of genes of an enzyme group participating in lipid metabolisms in a living body, and is therefore useful for treating and preventing various kinds of diseases related to obesity prevention and lipid metabolisms. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a composition for reducing internal fat, and when the composition for reducing internal fat of the present invention is ingested, the accumulation of visceral fat and the like is eliminated, and visceral fat-type obesity, which is said to be the cause of adult disease, is prevented. be able to.

  In recent years, eating habits have been enriched in Japan, and adult diseases such as arteriosclerosis and diabetes are rapidly increasing. It has been pointed out that these causes include obesity due to excessive intake of fat and saccharides and factors of lack of exercise even though the basal metabolic rate has decreased with aging. In particular, visceral fat-type obesity, in which fat accumulates in the viscera, has an extremely high risk of developing adult disease compared to subcutaneous fat-type obesity, in which fat accumulates subcutaneously. It is considered important to prevent. From this point of view, there is a demand for the development of substances and agents that eliminate the accumulation of visceral fat and the like, but the reality is that the purpose has not yet been fully achieved.

  Togedokoro has long been known to have nourishing tonic, anti-fatigue and muscle strengthening effects, and glucomannan obtained from Togedokoro has been known to have serum and liver lipid lowering effects (non-patented) Reference 1).

  The mechanism of action of glucomannan is a mechanism that wraps what is eaten in the stomach and suppresses digestion and absorption, and its physical properties are high in water swellability and have limited usage. In addition, application development of Tedokoro focusing on the high content of diosgenin has been carried out (Patent Document 1).

  In addition, tomorrow is known to have various health promoting effects, and chalcone derived from tomorrow has inhibition of 3-hydroxy-3-methylglutaryl-CoA reductase, which is the rate-limiting enzyme of cholesterol biosynthesis. It is known to be effective and effective in preventing hyperlipidemia (Patent Document 2).

JP 2007-27985 A International Publication No. 2005/074906 Pamphlet

Br. J. et al. Nutr. (British Journal of Nutrition), 2006, 96, 1021-1029

  However, the research and application development related to Tedokoro and tomorrow are not sufficient. It is desired to clarify the functions of Togekokoro and Tomorrow, which have not been known so far, and to use them.

Under the circumstances as described above, the present inventors have intensively researched components other than dietary fibers that are insoluble in water and difficult to use as a functional material, and have completed the present invention.
That is, the present invention
[1] A composition for reducing body fat, characterized by containing a substance derived from Togekokoro and / or a substance derived from tomorrow as an active ingredient,
[2] The composition for reducing body fat according to [1], wherein the Tedokoro-derived material is a processed Tedokoro product;
[3] The composition for reducing body fat according to [2], wherein the processed processed product is a powder of processed product.
[4] The composition for reducing body fat according to [2], wherein the processed product is a extract containing a fat-soluble toggedo-derived material
[5] The composition for reducing body fat according to [4], which is an extract obtained by extracting aqueous treatment ethanol from a heat-treated Tedokoro.
[6] The composition for reducing body fat according to [1], wherein the Tedokoro-derived substance is a diosgenin glycoside,
[7] The composition for reducing body fat according to [1], wherein the substance derived from tomorrow is a chalcone-containing substance,
[8] A food comprising the composition for reducing body fat according to any one of [1] to [7],
[9] A feed comprising the composition for reducing body fat according to any one of [1] to [7],
[10] A biological feeding agent comprising the composition for reducing body fat according to any one of [1] to [7],
[11] A method for raising an organism, comprising administering the composition for reducing body fat according to any one of [1] to [7] to the organism, and [12] a composition comprising a material derived from Togekokoro and a material derived from tomorrow ,
About.

  By ingesting the composition for reducing body fat of the present invention, gene expression of enzymes involved in lipid metabolism is improved, and as a result, body fat is reduced in living organisms. The composition for reducing internal fat according to the present invention has a remarkable effect in that it works well in the absence of exercise load.

The present invention is described in detail below.
The Tedokoro (scientific name Dioscorea esculenta) used in the present invention is a kind of yam cultivated mainly in the south of Kyushu, also called a potato.
In the composition of the present invention, any portion of the spider can be used, and there is no particular limitation. However, it is particularly preferable to use a rhizome (so-called potato).

  Tomorrow (scientific name: Angelica keiskei) used in the present invention is a large perennial of the celery family that is native to the Pacific coast, also called Hachijo-an. In the composition of the present invention, substances derived from any part of tomorrow may be used, and substances derived from fruits, seeds, seed coats, flowers, leaves, stems, roots, rhizomes and / or whole plants are used. Substances derived from leaves and stems can be preferably used.

  In a preferred embodiment of the present invention, a processed product of Tedokoro is used as the Tedokoro-derived material. Moreover, the processed product of tomorrow is used as a substance derived from tomorrow. The treated product is not particularly limited, but may be any material that has been subjected to some processing on the raw material, such as Tedokoro or tomorrow, and examples include a ground product, a dry powder, a pulverized product, a juice, a crushed product, and an extract. Is done.

  In the present invention, the ground product refers to a product obtained by grinding a plant body, and is usually prepared by using various automatic grinders or by manually grinding.

  In the present invention, the pulverized product and the crushed material refer to, for example, those obtained by cutting a plant body or those crushed after drying, and generally those having a large particle size are referred to as crushed materials, and those having a small particle size. Is called a pulverized product. In the present invention, the dry powder refers to a powder having a particle size smaller than that of the pulverized product or crushed product, and a method for producing such a dry powder includes, for example, drying rhizomes of Togedokoro and pulverizing. A method of obtaining a dry pulverized product derived from a powdered Tedokoro by pulverizing using a machine. In addition, as described in, for example, International Publication No. 2004/112817 pamphlet, tomorrow leaf extract extracted with water-containing ethanol is dried, and further pulverized using a pulverizer, whereby an ashitaba containing tomorrow leaf chalcone is obtained. A dry powder of the extract can be obtained. Further, a dry powder may be obtained by freeze pulverization or pulverization after freeze-drying.

  The crushed material is a material plant that has been crushed and generally has a larger tissue piece than the pulverized material, and can be produced, for example, by using a crusher. The squeezed liquid can be prepared by a known plant squeezing method. The obtained squeezed liquid can be used as it is, or solidified by means such as addition of excipients, spray drying, freeze drying and the like.

  As one aspect of the present invention, a composition containing a treated product obtained by drying a rhizome and then heat-treating it to insolubilize a viscous substance is exemplified. The heat treatment is carried out at a temperature of 50 ° C. or higher, preferably 60 to 150 ° C. for several seconds to 1 day, preferably 5 to 12 hours. The heat treatment is preferably dry heat treatment such as hot air treatment, roasting treatment, roasting treatment, or far-infrared treatment. The heat treatment insolubilizes viscous substances that hinder handling during pharmaceutical production and food and beverage production.

  An extract refers to a substance itself obtained through a process of performing an extraction operation on a raw material plant using an extraction solvent. The extraction can be performed as follows by a known extraction method. For example, after pulverizing or cutting the raw material, it can be carried out batchwise or continuously using a solvent. The aforementioned heat-treated product may be subjected to extraction. The extraction solvent for obtaining the extract is not particularly limited, but water, chloroform, ethanol, methanol, isopropyl alcohol and other alcohols, acetone, methyl ethyl ketone and other ketones, methyl acetate, ethyl acetate and other hydrophilic or An oleophilic solvent can be used, and it can be used alone or as a mixture as appropriate. An example of using the mixed solution as an extraction solvent is not particularly limited. For example, various aqueous solutions can be used. For example, 10 to 95%, preferably 15 to 90%, more preferably 20 to 85% alcohol. An aqueous solution can be used.

  The amount of the extraction solvent may be appropriately determined. Usually, the raw material plant is used in the form of the raw material plant as it is (for example, a raw plant if the raw plant is a raw plant), preferably 2 A 100-fold amount of extraction solvent may be used. The extraction temperature may be appropriately determined according to the purpose, but in the case of water extraction, it is usually preferably 4 to 130 ° C, more preferably 25 to 100 ° C. Moreover, when ethanol is contained in a solvent, the range of 4-60 degreeC is suitable from a safety viewpoint. The extraction time may be determined in consideration of the extraction efficiency. Usually, the raw material, the extraction solvent, and the extraction temperature are set so as to be preferably in the range of several seconds to several days, more preferably 5 minutes to 24 hours. Is preferred. The extraction operation may be performed, for example, while stirring or standing, and may be repeated several times as necessary.

  The extract can be used in the present invention as a concentrated solution or a solidified product as a solution dissolved in a solvent. A well-known means can be used for concentration and solidification.

  The extract prepared by the above operations was subjected to filtration, centrifugation, concentration, ultrafiltration, molecular sieving, and other treatments as necessary to concentrate the active ingredient having an action to reduce body fat. A treated product can be prepared. Moreover, the composition of this invention can also be made to contain 2 or more types of extracts obtained by the different extraction method from a raw material plant.

  Although the present invention is not particularly limited, the active ingredient of the composition of the present invention is exemplified by fat-soluble substances derived from Tedokoro. Examples of the fat-soluble substance include 60% (v / v) water-containing ethanol extract of heat-treated products of rhododendron rhizomes. A fat-soluble substance having the same effect as this extract can be used as the active ingredient of the present invention, and the use of a fat-soluble substance having the same effect as this extract is included in the present invention. A person skilled in the art can easily produce the fat-soluble substance from the disclosure of the present application.

  Moreover, although this invention is not specifically limited, the substance derived from tomorrow containing the chalcone is mentioned as an active ingredient of this invention. Examples of the substance include 55% (v / v) hydrous ethanol extract of tomorrow and its dry powder.

  The composition of the present invention can be produced by forming the above-mentioned processed product of togekokoro and / or processed product of tomorrow into an ingestible shape. Processed toggedokoro or processed tomorrow leaves may be used alone, combined with processed toggedokoro and processed tomorrow, or used in combination with other types of processed products. Also good. Since the body fat reducing effect of the Toledokoro-derived substance and the body fat reducing effect of the Asuka leaf-derived substance exhibit a synergistic effect, a composition containing the Togedocoro-derived substance and the Asuka leaf-derived substance is one of the preferred embodiments of the present invention. The composition of the present invention is preferably in a form that can be taken orally. In its production, it may be mixed and processed with a suitable solid or liquid carrier, preferably a pharmaceutically acceptable carrier. Furthermore, other components having a body fat reducing action and other physiological actions can be added. The shape is not particularly limited, and may be solid (including powdered and granular compositions) or liquid. The composition can be produced in an arbitrary shape by a known processing method. Although this invention is not specifically limited, For example, the processed product of Togedokoro and / or the processed product of tomorrow can be granulated by a well-known method, and can be used as a granular solid. The granulation method is not particularly limited, but rolling granulation, stirring granulation, fluidized bed granulation, air flow granulation, extrusion granulation, compression molding granulation, pulverization granulation, spray granulation, spray granulation or spray granulation Examples are grains. The treated product in powder form can be dissolved or suspended in a liquid such as water or alcohol to obtain a liquid, which can be used as the composition of the present invention. Furthermore, it is good also as a shape which is easy to take orally, such as tablet shape, granule shape, and capsule shape.

In addition, the composition of the present invention may further contain a substance known to have an effect of reducing body fat or visceral fat and / or an effect of improving lipid metabolism. For example,
(A) Togekokoro-derived material and / or tomorrow-derived material, and (B) Astaxanthin, Ercampule, Skipjack peptide, Garcinia, Kawara ketsumei, citrus fruit extract, kiwi peel extract, mushroom chitosan, conjugated linoleic acid, black soybean seed coat extract , Mulberry leaves, Ketchmeishi, Kotarahim, Rice Kefiran, Coleus Forskohlii, Sissasquad langlaris, Potato extract, Plant sterol, Vegetable oil emulsion (fabulous), Onion quercetin, Chia seed, Tsuruarame extract, DHA binding phospholipid, Neem , A composition containing at least one substance selected from the group consisting of fenugreek (coloroja), grape resveratrol, policosanol, maitake extract (glycrin), maca extract, mango ginger, and phospholipid It is also one aspect of the present invention.

  Moreover, this invention provides the foodstuff containing the processed material of this invention. Here, “containing” means containing, adding and / or diluting. The "containing" is an aspect that the active ingredient used in the present invention is contained in food, and the "addition" is an aspect in which the active ingredient used in the present invention is added to the raw material of the food. The “dilution” refers to an aspect in which food ingredients are added to the active ingredient used in the present invention. Such foods are also included in the composition of the present invention.

  There is no limitation in particular in the manufacturing method of the foodstuff containing the processed material of this invention. For example, blending, cooking, processing, and the like may be in accordance with those of general foods and can be produced by their production method, and the obtained food may contain the above-described composition of the present invention. There is no particular limitation on the type of food, for example, processed grain products, processed oil products, processed soybean products, processed meat products, marine products, dairy products, vegetables and the like, containing the active ingredient according to the present invention. Examples include processed fruit products, confectionery, alcoholic beverages, taste drinks, seasonings, spices, and other agricultural / forestry processed products, livestock processed products, and processed fishery products. The food may be in liquid form. That is, the food includes beverages.

  As another aspect of the present invention, an alcoholic beverage containing an active ingredient having a body fat reducing action derived from Tedokoro and / or an active ingredient having a body fat reducing action derived from tomorrow can be mentioned. The alcoholic beverage can be produced by immersing Tedokoro and / or tomorrow leaves in ethanol suitable for drinking or hydrous ethanol. Toggedokoro and tomorrow may be left as they are, or a ground product, a dry powder, a pulverized product, a crushed product, etc. may be prepared and immersed, or the heat-treated product may be immersed. Alcoholic beverages can also be produced by adding squeezed liquor or tomorrow leaves juice or extract to alcohol or hydrous alcohol. There is no particular limitation on the immersion time, but the immersion is usually 6 hours or more, preferably 12 hours to 1 year, more preferably 1 day to 3 months. When non-hydrated ethanol is used for the immersion, it may be adjusted to a desired concentration with water or an appropriate beverage or drinking water to obtain a beverage. As the water-containing ethanol, a water-diluted solution of food-grade ethanol, a commercially available alcoholic beverage, shochu, vodka, gin or the like may be appropriately selected and used. The ethanol concentration is not particularly limited, but it is preferably used within the range of 10% (v / v) to 35% (v / v). Needless to say, the water-containing ethanol used can be diluted with water or an appropriate beverage or drinking water so that the ethanol concentration is within a range suitable for drinking. Furthermore, in the production of beverages, components (sweeteners, acidulants, flavorings, fruit juices, thickeners, colorants, preservatives, antioxidants) used for the production of ordinary beverages can be blended. Furthermore, it is good also as an alcoholic beverage containing carbonic acid.

  The composition for reducing body fat of the present invention can be directly ingested as it is, or can be used in the form of a liquid, granule, tablet or the like using a known carrier or auxiliary agent. Furthermore, the composition for reducing body fat can be used after being added, mixed, contained and / or diluted in foods and beverages rich in lipids. And when using the composition for reducing body fat of the present invention for foods and beverages, the composition is manufactured through a heat treatment step, so it has thermal stability and is not used for water swelling. The advantage of being easy to do. Moreover, when using the composition for body fat reduction of this invention for a foodstuff, it is preferable to prepare so that it may become 0.1 mg-1g per meal.

  By ingesting the composition for reducing body fat of the present invention, body fat, particularly visceral fat is reduced in an individual organism. Furthermore, by ingesting the composition of the present invention, genes of enzymes involved in lipid metabolism, for example, PGC-1a (PPARγ coactivator-1), which is a transcriptional activator of genes related to energy metabolism, genes related to lipid metabolism PPAR-α (peroxisome proliferator-activated receptor-alpha), PPAR-δ (peroxisome proliferator-activated receptor-delta) that regulates the expression of fatty acid β-oxidation in fatty acids β oxidation in peroxisomes, ACO acyl that participates in fatty acid β oxidation in peroxisomes MCAD (medium chain acyl-CoA dehydrogenase) involved in β-oxidation, mitochondrial The expression level of UCP3 (uncoupling protein 3) involved in energy conversion and heat production of a increases. In addition, the concentration of adiponectin in the blood increases. From this, it is shown that the composition of the present invention exerts a fat reduction effect not by inhibiting fat absorption but by participating in the metabolic system.

  Since the composition of the present invention has a fat-reducing action, it can be used for diseases in which reduction of body fat is effective for treatment and prevention. Furthermore, as described above, the composition of the present invention activates the expression of genes of enzymes involved in lipid metabolism, so that the increase in the products of these genes is effective for improving symptoms and healing. The composition is effective. Examples of diseases for which the composition of the present invention is effective for treatment and prevention include metabolic syndrome, diabetes, hypertension, dyslipidemia, insulin resistance and the like.

  The present invention also provides a biological feed for reducing internal fat, comprising, adding and / or diluting the internal fat reducing composition of the present invention. Here again, “containing” means “containing, adding and / or diluting”. As another aspect, there is also provided a biological breeding agent comprising the composition for reducing internal fat of the present invention. Further, the present invention provides a method for raising an organism, comprising administering the composition for reducing body fat of the present invention or a feed containing the composition to the organism. The breeding method can be carried out by feeding or administering the aforementioned feed of the present invention and the biological breeding agent of the present invention to the organism.

  In the present specification, the organism is not limited, and examples thereof include farm animals, pet animals, and competition animals. Examples of farmed animals include domestic animals such as horses, cows, pigs, sheep, goats, camels and llamas, laboratory animals such as mice, rats, guinea pigs and rabbits, poultry such as chickens, ducks, turkeys and eagle birds, and fish. Examples of pet animals include dogs and cats. Examples of competition animals include race horses, race dogs, race camels, and the like. Examples of animal feed include feed for maintaining and / or improving physical condition. Examples of biological breeding agents include feed additives, beverage additives, and fish immersion agents.

  There is no limitation in particular in the manufacturing method of the feed of this invention, For example, what is necessary is just to make the composition for body fat reduction of this invention contained in the well-known feed suitable for the organism to breed. Also, the composition may be in accordance with general feed and should not affect the growth or health of the organism to be bred. The biological breeding agent may be prepared into a composition having an appropriate shape in accordance with the disclosure of the present specification, and may be in a form that can be directly administered to living organisms such as tablets, powders, injections, feeds and drinking water. It may be a shape that can be mixed. In the case of a feed additive, shapes suitable for mixing with feed such as powder, granules, pellets, and liquid are exemplified. The additive for beverage or the soaking agent for fish may be in a form that is soluble or suspendable in water.

  According to these inventions, the fat of an organism, for example, visceral fat is reduced, and the weight is not reduced. Since the aquaculture organism obtained by the breeding method of the present invention has reduced fat content and fat content in meat, it can provide meat with reduced risk of adult diseases and metabolic syndrome. That is, the present invention provides a method for modifying the flesh quality of organisms. Furthermore, the feed of the present invention is effective for preventing obesity and maintaining health of pet animals.

  Current livestock breeding is very intensive and grows in an environment with a low amount of exercise, but even in such an environment, high-quality meat with low fat can be produced by ingesting the feed of the present invention. Pet food is also provided that does not require low-energy or reduced food.

  The present invention also provides an anti-obesity composition comprising yam, a processed product thereof, tomorrow, and / or a processed product thereof as an active ingredient.

  An example of a yam is Tedokoro. Examples of the treated product include a dried product, for example, a dry heat-treated product, for example, a far-infrared-treated product or a fat-soluble yam-derived material, for example, a fat-soluble toggedokoro-treated product, for example, a water-containing ethanol extract. Compositions are provided.

  Further, according to the present invention, a composition for activating intracellular energy consumption or a composition for promoting intracellular energy consumption comprising a processed product of swordfish as an active ingredient is provided. A composition for promoting the expression of a gene related to intracellular energy consumption is provided. Examples of the processed product of the toggedoro include a treated product containing diosgenin glycoside, and a composition for activating intracellular energy consumption or a composition for promoting intracellular energy consumption, which comprises the processed product as an active ingredient. In addition, as an aspect of the present invention, a composition containing a diosgenin glycoside is exemplified. In the present embodiment, the composition is limited to the genus Tongokoro as long as it is a composition containing a diosgenin glycoside. It is not something.

  Examples of intracellular energy consumption-related genes or intracellular energy consumption-related genes include genes expressed when fatty acids are used as an energy source, such as genes involved in β-oxidation of fatty acids, such as the MCAD gene and ACO gene described above. In addition, expression of transcription factors that regulate the expression of these genes, such as the aforementioned PGC-1a gene, PPAR-α gene, and PPAR-δ gene, is also promoted. That is, by ingesting the composition of the present invention, it has been found that fat is decomposed to become fatty acid and consumed as an energy source in the cells, and the body fat is reduced, resulting in exerting an anti-obesity effect. The composition of the present invention has a significant effect as an anti-obesity composition by having these actions without accompanying exercise load.

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

Example 1 Reduction of fat by administration of treated product of togekokoro (1) Preparation of heat-treated product of togekokoro Dioscorea esculenta sliced to about 5 mm was dried by heating at 70 ° C. for about 8 hours. Thereafter, the dried product was further subjected to heat treatment (135 to 140 ° C., 15 seconds) and pulverized to obtain a powdered twig.

(2) Preparation of Tedokoro 60% (v / v) hydrous ethanol extract 800 g of the above Tedokoro heat-treated powder was added to 8 L of 60% hydrous ethanol while stirring, and the mixture was stirred for 24 hours at 25 ° C. using a stirrer. Suction filtration was performed using a funnel (filter paper No. 2 manufactured by Whatman). The filtrate was dried with an evaporator to obtain 98.9 g of a 60% aqueous ethanol extract.

(3) The body fat reducing effect of the heat-treated powder of swordfish and water-containing ethanol extract.
Male BALB / c mice were purchased from Japan SLC and used for experiments from 7 weeks of age after preliminary breeding. A test meal was prepared by mixing the powdered heat treated powder prepared in Example 1- (1) with standard feed CE-2 (manufactured by CLEA Japan, Inc.) at a rate of 1% (w / w). In addition, a test meal in which the 60% aqueous ethanol extract prepared in Example 1- (2) was mixed with the standard powder sample CE-2 was also used. Prior to administration of the test meal, individuals capable of swimming for a predetermined time or more were selected in advance and equally distributed to each exercise group and non-exercise group.

  After that, a test group that freely ingests the test meal and a control group that freely ingests the standard feed CE-2 are set, and each exercise group has the third day of the start of the test meal administration and performs swimming exercise once a week. It was. The swimming exercise time was defined as the time from when the mouse was loaded with a weight of 1% of the body weight and forced swimming was started, and the head was completely submerged in water for 5 seconds after fatigue. On the next day after the exercise exercise was performed until the head was completely submerged in water for 5 seconds after fatigue at 8 weeks after administration of the test food, the mice were lethal and lethal under anesthesia. Non-motor group mice were treated in the same manner. The fat around the testis of each group of mice was excised and weighed. The average of 9-15 animals in each group is shown in Table 1. Compared with the control group, the amount of fat decreased in the administration group of 1% (w / w) heat-treated powder of Togedokoro and 60% aqueous ethanol extract. There was no significant difference between the groups regarding the weight of the mice.

  As shown in Table 1, regardless of the presence or absence of exercise during the breeding period, a decrease in the amount of fat was observed in the Tedokoro heat-treated powder (and 60% aqueous ethanol extract) administration group. From this result, it was found for the first time that the processed product of Tedokoro exerted a body fat reducing effect. Furthermore, 60% hydrous ethanol extract exhibited a stronger effect than heat-treated powder.

Example 2 Reduction of Free Fatty Acids in Blood by Ingestion of Togekokoro Powder In the experiment described in Example 1, blood of a non-exercise group of mice was collected after completion of ingestion, and the concentration of free fatty acid in serum was measured by NEFA C-Test Wako ( Wako Pure Chemical Industries, Ltd.). The results are shown in Table 2. In the Tedokoro powder intake group, a decrease in the free fatty acid concentration in the blood was observed compared to the control group.

Example 3 Influence on various gene expression by ingestion of powdered tortilla in liver and skeletal muscle In the experiment described in Example 2, liver and skeletal muscle (gastrocnemius) were extracted from a non-exercise group of mice that had been exsanguinated to death. Using RNAiso (manufactured by Takara Bio Inc.), a total RNA aqueous solution was prepared from the liver and skeletal muscle according to the description in its instruction manual.

  Reverse transcription reaction and real-time PCR are performed using SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (manufactured by Takara Bio Inc.), and the mRNA expression level is determined by Thermal Cycler Dice (registered trademark) Real Time System (Takara Bio). The measurement was performed using The β-actin gene in the liver and the GAPDH (glyceraldehyde-3 phosphate dehydrogenase) gene in the skeletal muscle were used as the correction genes, and the mRNA expression level in the group with the staghorn powder intake was calculated as a relative ratio to the control group.

  The base sequences of the primer pairs used in the measurement of the expression level of mRNA transcribed from each gene of PGC-1a, PPAR-α, ACO, MCAD, UCP3, β-Actin, and GAPDH are shown in SEQ ID NOs: 1 and 2, 3 and 4, SEQ ID Nos. 5 and 6, SEQ ID Nos. 7 and 8, SEQ ID Nos. 9 and 10, SEQ ID Nos. 11 and 12, and SEQ ID Nos. 13 and 14, respectively.

  The measurement results are shown in Tables 3 and 4 below. That is, Tables 3 and 4 show the rate of increase in mRNA expression in each organ of mice administered with Tedokoro, and PGC-1a, PPAR-α, ACO, and MCAD mRNA expression in the liver by ingestion of Tedokoro powder. The amount of mRNA expression of PGC-1a and UCP3 increased in skeletal muscle.

Example 4 Fat burning action by 60% ethanol extract of Tedokoro (1) Preparation of 60% ethanol extract of Tedokoro To 100 g of Tedokoro heat-treated powder prepared in Example 1, 100 mL of 60% ethanol was added and stirred. Extraction was performed at 25 ° C. for 24 hours. The extract was subjected to suction filtration and then concentrated to dryness. A solution obtained by dissolving the dried product in 10 mL of a 60% DMSO solution was used as a Tedokoro 60% ethanol extract.

(2) Enhancement of expression level of fatty acid metabolism-related genes in hepatocytes Mouse liver-derived Hepa1c1c7 cells are contained in Dulbecco's modified Eagle medium (Sigma, D5796) containing 10% fetal bovine serum and 1% Penicillin-Streptomycin. ^The suspension was suspended at ⁵ cells / mL, and 2 mL was added to each well of a 12-well plate, followed by culturing overnight at 37 ° C. in the presence of 5% carbon dioxide gas. Thereafter, the medium was replaced with a fresh medium, and the well-prepared 60% ethanol extract of Tedokoro prepared in Example 4- (1) was added to each well so as to have a final concentration of 0.05%, 0.1%, or 0.2%. Was added as a test substance and cultured for 24 hours. As a negative control, 60% dimethyl sulfoxide (DMSO) addition was set.

  After completion of the culture, the medium was removed, 0.5 mL of RNAiso (manufactured by Takara Bio Inc.) was added, and the cells were collected in a 1.5 mL Eppendorf tube. After leaving at room temperature for 5 minutes, 0.1 mL of chloroform was added and shaken well until it became milky white. After standing again at room temperature for 5 minutes, the mixture was centrifuged at 10,000 rpm for 15 minutes at 4 ° C., and the supernatant was transferred to another Eppendorf tube. To this supernatant, 0.25 mL of isopropanol was added, mixed well, and allowed to stand at room temperature for 10 minutes. Centrifugation was performed at 10,000 rpm for 10 minutes at 4 ° C., and the resulting precipitate was washed with 0.5 mL of 75% ethanol. After centrifugation at 10,000 rpm for 5 minutes at 4 ° C., the precipitate was dried. The precipitate was dissolved with 20 μL of water for injection to obtain a total RNA aqueous solution. Reverse transcription reaction and real-time PCR were performed in the same manner as in Example 3. As a control, real-time RT-PCR using a primer pair specific to the β-actin gene was performed. All measurements were performed in triplicate.

  The results are shown in Table 5 below. That is, Table 5 shows the ratios of mRNA expression levels of PGC-1a and ACO to negative control cells in hepatocytes to which Tedokoro 60% ethanol extract was added. In addition, the activity of inducing the expression of ACO mRNA was observed.

(3) Enhancement of expression level of fatty acid metabolism-related gene in muscle cells C2C12 cells were placed in 10% fetal bovine serum, 1% Penicillin-Streptomycin, low glucose-containing Dulbecco's modified Eagle medium (Sigma, D6046) at 8 × 10 ^After suspending at ⁴ / mL and adding 2 mL to each well of a 12-well plate, the cells were cultured at 37 ° C. for 3 days in the presence of 5% carbon dioxide gas.

  Next, the medium was changed to Dulbecco's modified Eagle medium containing 2% horse serum, 1% penicillin-streptomycin, and low glucose, and cultured for 1 week while changing the medium every 2 to 3 days to differentiate C2C12 cells into muscle cells. It was. Thereafter, the medium was replaced with a fresh medium, and the Tedokoro extract prepared in Example 4- (1) was added to the wells to a final concentration of 0.05%, 0.1%, and 0.2%. Incubate for hours. As a negative control, 60% dimethyl sulfoxide (DMSO) addition was set.

  After completion of the culture, the expression level of PGC-1a mRNA was measured by the same method as in Example 4- (2). As a control, real-time RT-PCR using a primer pair specific to the GAPDH gene was performed.

  The results are shown in Table 6 below. That is, Table 6 shows the ratio of the expression level of PGC-1a mRNA relative to the negative control cells in the cells to which Tedokoro 60% ethanol extract was added. Activity to induce expression was observed.

Example 5 Isolation and identification of expression enhancing component of fatty acid metabolism-related gene from Tedokoro (1) Fractionation of Tedokoro 60% ethanol extract 100 g of 60% ethanol for 10 g of Tedokoro heat-treated powder prepared in Example 1 And extracted with stirring at 25 ° C. for 24 hours. The extract was subjected to suction filtration, concentrated to dryness, and the dried product was dissolved in 10 mL of 60% ethanol. Of the resulting lysate, 2.5 mL was subjected to HPLC for fractionation. The HPLC conditions are shown in Table 7.

  Each fraction was fractionated for each retention time shown in Table 8 using UV 205 nm absorption as an index.

  Next, each fractionated fraction was concentrated and dried, and the dried product was dissolved in 1 mL of 60% ethanol.

(2) Measurement of activity of fraction of 60% ethanol extract of Togedokoro About the fraction of 60% ethanol extract of Togedokoro obtained in Example 5- (1), the same method as in Example 4- (2) The activity of inducing mRNA expression of fatty acid metabolism-related genes was confirmed. As a result, fractions 7, 8, 9, and 10 were found to have the activity of inducing the expression of PGC-1a and ACO mRNA.

(3) Structural analysis of fractions 7 to 10 The molecular weight of fractions 7 to 10 was measured using a mass spectrometer (API300, manufactured by Applied Biosystems). As a result, both fractions 7 and 8 had a molecular weight of 1014, fraction 9 had a molecular weight of 868, and fraction 10 had a molecular weight of 884.

  Next, the fractions 7 to 10 were acid hydrolyzed under the following conditions. To each 0.1 mL of fractions 7 to 10, 0.2 mL of 80% ethanol was added and stirred, and then 0.2 mL of 10 M sulfuric acid was added and acid hydrolysis was performed at 100 ° C. for 1 hour. After completion of the reaction, 3 mL of water was added, and liquid-liquid distribution was performed with 2 mL of t-butyl methyl ether (BME) to recover the BME layer. For the aqueous layer, the liquid-liquid partition with 2 mL of BME was repeated twice more. Next, the BME layer obtained by a total of three liquid-liquid distributions was mixed, and 1 mL of 1M aqueous sodium hydroxide solution was added thereto and stirred, and then the BME layer was recovered. To the recovered BME layer, 1 ml of 1M sodium hydroxide aqueous solution was added again and stirred, and then the BME layer was recovered. 1 mL of water was added to the obtained BME layer and stirred, and then the BME layer was collected and concentrated to dryness. The dried product was dissolved in 1 mL of ethanol. The acid hydrolyzate thus obtained was subjected to HPLC analysis under the conditions shown in Table 9 and compared with a commercially available diosgenin standard substance (manufactured by Wako Pure Chemical Industries, Ltd.). As a result, since diosgenin was detected from all the fractions, it became clear that fractions 7 to 10 were diosgenin glycosides.

  Further, the fraction 9 was subjected to HPLC analysis under the conditions shown in Table 10 and compared with a commercially available dioscin standard substance (manufactured by ChromaDex). As a result, it was revealed that fraction 9 was dioscin.

(4) Enhancement of expression level of fatty acid metabolism-related genes in fractions 7 to 10 Fraction fraction of diosgenin glycoside prepared in Example 5- (1) in Hepa1c1c7 cells by the same method as in Example 4- (2) 7 to 10 were added so that the final concentration of the diosgenin glycoside was 2.5 to 10 μM and cultured for 24 hours. In addition, the DMSO addition category was set as a negative control. After completion of the culture, the expression levels of PGC-1a and ACO mRNA were measured by the same method as in Example 4- (2). The results are shown in Table 11. That is, Table 11 shows the mRNA expression ratio of PGC-1a and ACO relative to the negative control cells in the cells to which Togekokoro-derived diosgenin glycosides were added. PGC-1a and ACO that are prominent in diosgenin glycosides are shown. The activity of inducing the expression of mRNA was observed.

(5) Enhancement of expression level of fatty acid metabolism-related gene in diosgenin glycoside fraction 8 Diosgenin glycoside fraction prepared in Example 5- (1) on Hepa1c1c7 cells by the same method as in Example 4- (2) Minute fraction 8 was added so that the final concentration of the diosgenin glycoside was 2 to 4 μM and cultured for 24 hours. Further, the diosgenin glycoside fraction 8 prepared in Example 5- (1) was applied to the C2C12 cells differentiated into muscle by the same method as in Example 4- (3), and the final concentration of diosgenin glycoside was 1. It added so that it might become -2micromol, and cultured for 24 hours. For any cell, a DMSO addition category was set as a negative control. After completion of the culture, real-time RT-PCR was performed in the same manner as in Example 4- (2) except that a primer pair specific to the PPAR-δ gene was used as a primer, and the expression level of PPAR-δ mRNA was measured. . The results of Hepa1c1c7 are shown in Table 12, and the results of C2C12 are shown in Table 13. In addition, the base sequences of primer pairs used for measurement of the expression level of mRNA transcribed from the PPAR-δ gene are shown in SEQ ID NOs: 15 and 16, respectively.

  That is, Tables 12 and 13 show PPAR-δ mRNA expression ratios relative to the negative control cells in the cells to which the tosedokoro-derived diosgenin glycoside was added. Activity to induce expression of mRNA was observed.

(6) Activity comparison between diosgenin glycoside and diosgenin Final concentration of diosgenin glycoside fraction fraction 8 and diosgenin prepared in Example 5- (1) on Hepa1c1c7 cells in the same manner as in Example 4- (2) It added so that it might become 1-4 micromol, and cultured for 24 hours. In addition, the division of DMSO addition was set as a negative control for diosgenin glycosides, and DMF was set as a negative control for diosgenin. After completion of the culture, the expression levels of PGC-1a and ACO mRNA were measured by the same method as in Example 4- (2). The results are shown in Table 14.

  That is, Table 14 shows the mRNA expression level ratios of PGC-1a and ACO relative to the negative control cells in the cells to which diosgenin glycosides and diosgenin were added. Although activity to induce expression of mRNA was observed, diosgenin showed almost no such activity. From this result, it became clear that the component which has the activity which reduces the body fat contained in a togedokoroko processed material is a diosgenin glycoside.

Example 6 Manufacture of soaked liquor using heat-treated powder (1)
Example 1-100 g of Tedokoro heat-treated powder prepared in (1) is placed in a wide-mouthed jar, 900 ml of white Takara 35% (Takara Shuzo Co., Ltd.) is poured and aged for 1 month, and soaked liquor using Tedokoro heat-treated powder is produced. did.

Example 7 Manufacture of immersion liquor using heat-treated powder (2)
100 g of heat-treated powder of Tedokoro prepared in Example 1- (1) was placed in a wide-mouthed bottle, and 900 mL of a 60% aqueous ethanol solution for food was poured and extracted for one week. The extract was diluted two-fold with tap water to produce a soaked liquor using Tedokoro heat-treated powder.

Example 8 Fat reduction due to tomorrow The subject of metabolic syndrome who has a waist circumference of 85 cm or more and a woman of 90 cm or more, and middle-aged (35 to 65 years old) reserve men and women (7 men, 2 women) Asitaba chalcone-containing Ashitaba extract powder (prepared as described in Example 12 of International Publication No. 2004/112817 pamphlet) 100 mg per package in Asahoba Aojiru (Takara Bio Inc., 1 package 3 g) The product (containing 6 mg of tomorrow leaf chalcone per packet) was dissolved in water and ingested twice a day for 8 weeks. As evaluation items, body weight, BMI, body fat percentage, abdominal fat area by CT scan, blood adiponectin, and blood LDL-cholesterol concentration were measured. The results are shown in Table 15. Informed consent was obtained from all subjects.

  That is, Table 15 shows the results of examination of various evaluation items for fat reduction due to tomorrow. The intake of tomorrow leaf green juice showed a decrease in abdominal fat area, and particularly a decrease in abdominal visceral fat area was remarkable. It was. In addition, body weight, BMI, body fat percentage, and blood LDL-cholesterol concentration also decreased. Furthermore, blood adiponectin increased with ingestion.

Example 9 Enhancement of expression level of fatty acid metabolism-related gene by co-addition of tomorrow chalcone and tomorrow chalcone and diosgenin glycoside (1) Preparation of tomorrow leaf chalcone Example 1 of WO 2005/074906 and According to the method described in Example 2, xanthoangelol (XA) and 4-hydroxyderricin (4-HD) were prepared from tomorrow.

(2) Enhancement of expression level of fatty acid metabolism-related gene by tomorrow leaf chalcone and diosgenin glycoside Tomorrow leaf chalcone 4HD and XA prepared in Example 9- (1) instead of Togekokoro 60% ethanol extract, and implementation Example 5 The point of adding the diosgenin glycoside fraction 8 prepared in (1) as a test substance to Hepa1c1c7 cells at the final concentrations shown in Tables 16 and 17 and the DMSO addition category as a negative control are set. Except for this point, the activity of inducing the expression of PGC-1a mRNA indicated by the test substance was confirmed by the same method as in Example 4- (2). The results are shown in Table 16 and Table 17. That is, Tables 16 and 17 show the ratio of the expression level of PGC-1a mRNA relative to the negative control cells in the cells to which the tosedokoro-derived diosgenin glycoside and tomorrow-derived chalcone were added alone or in combination. The activity of inducing the expression of PGC-1a mRNA was also observed with leaf chalcone alone, and the activity of further inducing the expression of PGC-1a mRNA by co-addition of tomorrow leaf chalcone and diosgenin glycoside rather than the addition of alone Was recognized. This result shows the synergistic effect of the body fat reducing effect of the toggedokoro-derived substance and the body fat reducing effect of the tomorrow leaf-derived substance.

  According to the present invention, a composition for reducing body fat containing an active ingredient derived from Togekokoro and / or an ingredient derived from tomorrow is provided, and an anti-obesity drug or a food or drink is provided using the composition.

SEQ ID NO: 1; Primer to amplify the cDNA fragment of mouse PGC-1 gene.
SEQ ID NO: 2; Primer to amplify the cDNA fragment of mouse PGC-1 gene.
SEQ ID NO: 3; Primer to amplify the cDNA fragment of mouse PPAR-alpha gene.
SEQ ID NO: 4; Primer to amplify the cDNA fragment of mouse PPAR-alpha gene.
SEQ ID NO: 5; Primer to amplify the cDNA fragment of mouse ACO gene.
SEQ ID NO: 6; Primer to amplify the cDNA fragment of mouse ACO gene.
SEQ ID NO: 7; Primer to amplify the cDNA fragment of mouse MCAD gene.
SEQ ID NO: 8; Primer to amplify the cDNA fragment of mouse MCAD gene.
SEQ ID NO: 9; Primer to amplify the cDNA fragment of mouse UCP3 gene.
SEQ ID NO: 10; Primer to amplify the cDNA fragment of mouse UCP3 gene.
SEQ ID NO: 11; Primer to amplify the cDNA fragment of mouse beta-actin gene.
SEQ ID NO: 12; Primer to amplify the cDNA fragment of mouse beta-actin gene.
SEQ ID NO: 13; Primer to amplify the cDNA fragment of mouse GAPDH gene.
SEQ ID NO: 14; Primer to amplify the cDNA fragment of mouse GAPDH gene.
SEQ ID NO: 15; Primer to amplify the cDNA fragment of mouse PPAR-delta gene.
SEQ ID NO: 16; Primer to amplify the cDNA fragment of mouse PPAR-delta gene.

Claims (12)

  A composition for reducing body fat, comprising a substance derived from Togekokoro and / or a substance derived from tomorrow as an active ingredient.   The composition for reducing body fat according to claim 1, wherein the toggedokoro-derived substance is a processed toggedokoro product.   The composition for reducing internal fat according to claim 2, wherein the processed product is a heat-treated powder of Togedokoro.   The composition for reducing body fat according to claim 2, wherein the processed product is a extract containing a fat-soluble material.   The composition for reducing body fat according to claim 4, wherein the processed product is a product obtained by extraction with water-containing ethanol from a heat-treated product.   The composition for reducing body fat according to claim 1, wherein the Todokoro-derived material is a diosgenin glycoside.   The composition for reducing body fat according to claim 1, wherein the substance derived from tomorrow is a chalcone-containing substance.   A food comprising the composition for reducing body fat according to any one of claims 1 to 7.   A feed comprising the composition for reducing body fat according to any one of claims 1 to 7.   A biological breeding agent comprising the composition for reducing body fat according to any one of claims 1 to 7.   A method for raising an organism, comprising administering the composition for reducing body fat according to any one of claims 1 to 7 to the organism.   A composition comprising a material derived from Togekokoro and a material derived from tomorrow.