JP5340520B2 - Weight gain inhibitor - Google Patents

Description

The present invention relates to a weight gain inhibitor having a weight gain inhibitory action.

Conventionally, it has been reported that citrus-derived polyphenol has an antioxidant effect, a flavor deterioration preventing effect, an ultraviolet ray absorbing effect, and the like (for example, see Patent Documents 1 to 3).
JP 9-48969 A JP 2001-61461 A Japanese Patent Laid-Open No. 2001-200288

The present invention was made by finding out that, as a result of intensive studies by the present inventors, a hydrophilic component containing a polyphenol obtained from citrus has a useful effect other than the conventionally reported action. Is. The purpose is to provide a weight gain inhibitor that can exert a high weight gain inhibitory effect.

In order to achieve the above object, the weight gain inhibitor according to claim 1 adsorbs a hydrophilic component in the extract to an adsorption resin with respect to an extract obtained using water from a lemon juice residue. Ingredients eluted with an aqueous ethanol solution are contained as active ingredients, and the ingredients eluted with the aqueous ethanol solution contain eriocitrin, hesperidin, nariltin, and diosmine as the main components to suppress weight gain. The gist is intended. Since this weight gain inhibitor has a hydrophilic component containing polyphenol as an active ingredient, a high weight gain inhibitory effect is exhibited due to the hydrophilic component.

The weight gain inhibitor according to claim 2 is eluted with an aqueous ethanol solution by adsorbing a hydrophilic component in the extract to an adsorbent resin with respect to an extract obtained using water from lemon juice residue. It is an object of the present invention to contain an ingredient as an active ingredient and to have an action of increasing the expression of at least one gene selected from a fatty acid CoA ligase gene and an acyl CoA oxidase gene in a weight gain inhibitor intended to suppress weight gain. To do.

The body weight gain inhibitor, since the relative dosing individuals, has the effect of increasing the expression of lipid metabolism genes of the fatty acid combustion system, enhance the decomposition of fatty acids, resulting in higher body weight gain Demonstrate the effect.

According to the weight gain inhibitor of the present invention, a high weight gain inhibitory effect can be exhibited.

Hereinafter, an embodiment in which the present invention is embodied in an anti-obesity agent will be described.
The anti-obesity agent (weight gain inhibitor) of this embodiment contains a hydrophilic component derived from citrus fruits as an active ingredient. This anti-obesity agent acts on the individual administered with the anti-obesity agent due to the action of the active ingredient, and suppresses the increase in body weight (the effect of reducing body weight) or the increase in the body fat percentage (increase in body fat percentage). It exerts an anti-obesity effect through a lowering action).

  Furthermore, this anti-obesity agent suppresses the biosynthesis of fatty acids and cholesterol through the action of lowering the expression of lipid metabolism enzyme genes in the fatty acid synthesis system and cholesterol synthesis system for the administered individual. In addition, this anti-obesity agent enhances the degradation of fatty acids in the administered individual through the action of increasing the expression of the fatty acid burning system lipid metabolism enzyme gene. Examples of fatty acid synthesizing lipid metabolism enzyme genes include ATP citrate lyase gene, acetyl CoA carboxylase gene, and fatty acid synthase gene. Examples of the cholesterol metabolism enzyme gene of cholesterol synthesis system include HMG-CoA reductase (Hydroxymethylglutaryl-CoA reductase) gene. Examples of the fatty acid combustion lipid metabolism enzyme gene include a fatty acid CoA ligase gene and an acyl CoA oxidase (ACO) gene.

  In addition, this anti-obesity agent can cause a decrease in blood neutral fat (triglyceride; TG) concentration and a decrease in blood cholesterol concentration in individuals administered with the anti-obesity agent. In the reduction of the blood cholesterol level, the high density lipoprotein (HDL) cholesterol level is increased while the low density lipoprotein (LDL) cholesterol level is significantly reduced. Resulting in a reduction in total cholesterol. For this reason, the antiobesity agent of this embodiment is also useful for prevention and treatment of hyperlipidemia, arteriosclerosis, heart disease and the like.

  The hydrophilic component contains polyphenol, and preferably contains polyphenol as a main component. The polyphenol as a main component means that the solid content in the hydrophilic component is higher than that of any other component, and the polyphenol is preferably 30 wt% or more, more preferably 50 wt%. % Or more. The hydrophilic components derived from lemon, lime and sudachi also contain a high amount of eriocitrin.

  The said hydrophilic component is highly contained in the polar solvent extract obtained by carrying out the polar solvent extraction of the raw material which consists of a citrus fruit or its component. As the polar solvent, lower alcohols such as methanol, ethanol, isopropanol, water, glycerin, glacial acetic acid and the like can be used, and water or ethanol is preferably used when a hydrophilic component is contained in a food or drink. Water is particularly preferably used because of its low cost. These listed polar solvents may be used alone or in combination of two or more. In the polar solvent extract, impurities such as pectin are simultaneously contained in addition to the hydrophilic component. For this reason, as a polar solvent extract, what removed impurities, such as pectin, is used as needed.

  The polar solvent extract is obtained by carrying out an extraction process in which citrus fruits or constituents thereof are used as raw materials, and the raw materials are stirred or left standing while immersed in a polar solvent. Examples of citrus fruits include lemon, lime, grapefruit, sudachi, yuzu, navel orange, Valencia orange, sour orange, hassaku, Wenzhou orange, Iyokan, Daidai, Kabosu and Ponkan. These citrus fruits may be used alone as raw materials, or two or more kinds may be used as raw materials in combination. Moreover, you may use the hybrid | sebration kind of these listed multiple types of citrus as a raw material. As the raw material used in the extraction step, since the polyphenol content in the obtained polar solvent extract is easily increased, citrus citrus fruits such as lemon, lime, sudachi, yuzu, daidai, kabosu are preferably used. Lemon, lime and sudachi are used, particularly preferably lemon.

  In these citrus fruits, examples of the fruit component include fruit skin, fruit juice, gills, saury and seeds. Among these components, since the polyphenol content in the obtained polar solvent extract can be easily increased, fruit juice and fruit skin are preferably used, and more preferably fruit skin is used. Moreover, as a raw material used in the extraction step, the fruit may be used as it is, but in order to save the labor required for production and to make effective use of the fruit, the juice residue after squeezing the fruit juice from the fruit is used. Most preferred. This squeezed residue contains fruit skin, scabbard, part of sasa, seeds, and a very small amount of fruit juice that could not be squeezed.

  In the extraction step, after the hydrophilic component contained in the raw material is transferred into the polar solvent, solid-liquid separation is performed to separate the raw material and the polar solvent. By this solid-liquid separation, a liquid polar solvent extract in which a hydrophilic component is dissolved in a polar solvent is obtained. For the solid-liquid separation, a known separation method such as centrifugation or membrane separation can be employed. After solid-liquid separation, the liquid polar solvent extract is concentrated and dried to obtain a powdery polar solvent extract. For concentration and drying of the polar solvent extract, known vacuum concentration and vacuum drying are employed. In addition, it is preferable that the extraction time in an extraction process is 2 hours or more.

  The polar solvent extract after solid-liquid separation is preferably subjected to a purification step for removing impurities and increasing the content of hydrophilic components. In this purification step, after applying an adsorption treatment in which a liquid polar solvent extract is applied to the adsorption resin to adsorb the hydrophilic component to the resin, a washing treatment for washing the adsorption resin is performed, and then the hydrophilic resin is removed from the adsorption resin. An elution process is performed to elute the components. As the adsorption resin, a resin capable of adsorbing a hydrophilic component is used. Examples of the material of the adsorption resin include styrene synthetic resins and acrylic synthetic resins. Specifically, Duolite S-861 (Rohm and Haas) can be used.

The adsorption process is a process of applying a polar solvent extract to the adsorption resin packed in the column, and is performed to adsorb the hydrophilic component in the polar solvent extract onto the adsorption resin.
The washing treatment is a treatment for washing the adsorption resin by flowing a washing solvent through the column after the adsorption treatment, and is performed to remove many impurities other than the hydrophilic component from the adsorption resin. As the washing solvent, a solvent capable of maintaining the adsorption state of the hydrophilic component with respect to the adsorption resin is used. Preferably, the polar solvent is used, and more preferably water is used. This washing process utilizes the fact that the elution rate of the hydrophilic component with respect to the washing solvent is slower than the elution rate of impurities. This washing solvent may be flowed through the column in an unheated state, but in order to efficiently remove impurities other than hydrophilic components, it may be flowed through the column while being heated to 40 to 100 ° C. preferable.

  The elution process is a process for eluting the hydrophilic component adsorbed on the adsorption resin by flowing an elution solvent through the column after the washing process, and is performed to recover the hydrophilic component from the column. By this elution treatment, an eluate in which the hydrophilic component is dissolved in the elution solvent is obtained. As an elution solvent, alcohols such as ethanol, organic solvents such as acetone, hexane, chloroform, glycerin, glacial acetic acid, and water can be used, and an aqueous ethanol solution is preferable when a hydrophilic component is contained in a food or drink. used. These enumerated solvents may be used alone or in combination of two or more. The eluate obtained by the purification step (elution treatment) can be powdered by concentrating and drying by a known method.

  The anti-obesity agent of this embodiment can be used by being added to pharmaceuticals, quasi drugs and the like. In this case, an anti-obesity effect, a blood TG reduction effect, and a blood cholesterol reduction effect can be exhibited, and a therapeutic effect and a preventive effect such as hyperlipidemia, arteriosclerosis, and heart disease can be exhibited. In these pharmaceuticals and quasi-drugs, the content of the hydrophilic component is preferably 0.15 to 30% by weight in order to exhibit sufficient efficacy and effects. In addition, in medicines and quasi-drugs, the intake of hydrophilic components varies depending on symptoms, age, weight, etc., but is preferably 0.5 to 100 g per day for an adult. In the case of a dwarf, half the amount for an adult is a guide.

  Moreover, the antiobesity agent of this embodiment can also be added and used for food-drinks. Food and drink products include food preparations such as health foods in addition to beverages such as drinks and foods such as cookies. In this case, the anti-obesity effect, the blood TG reduction effect, and the blood cholesterol reduction effect can be exhibited, and the prevention effects such as hyperlipidemia, arteriosclerosis, and heart disease can be exhibited.

  The content of the hydrophilic component in the food or drink is preferably 0.05 to 50% by weight. If the content of the hydrophilic component contained in the food / beverage product is less than 0.05% by weight, it is difficult to exert the above effect, and conversely if it exceeds 50% by weight, it is uneconomical. This food or drink is preferably taken orally in several divided doses per day. In food and drink, the intake amount of the hydrophilic component varies depending on age, weight, etc., but is preferably 0.5 to 2000 g per day for an adult. If the daily intake of food and drink is less than 0.5 g, it will be difficult to exert the above effect, and conversely if it exceeds 2000 g, it will be uneconomical. In the case of a dwarf, half the amount for an adult is a guide.

The effects exhibited by the embodiment will be described below.
-The antiobesity agent of this embodiment contains the hydrophilic component derived from the fruit of citrus fruits as an active ingredient. Since the hydrophilic component contains a high content of polyphenols that exhibit a high anti-obesity effect, the anti-obesity agent has an effect of suppressing body weight gain and body fat percentage for individuals administered with the anti-obesity agent. High anti-obesity effect can be exerted through an increase suppressing action of

  -The anti-obesity agent of this embodiment can suppress the biosynthesis of fatty acids and cholesterol through the action of reducing the expression of lipid metabolism enzyme genes in the fatty acid synthesis system and cholesterol synthesis system to the administered individual. it can. Furthermore, this anti-obesity agent can enhance the degradation of fatty acids in the administered individual through the action of increasing the expression of the fatty acid combustion system lipid metabolism enzyme gene. Therefore, this anti-obesity agent can exhibit a high anti-obesity effect at the individual level as a result of such expression control at the gene level.

  -The anti-obesity agent of this embodiment causes a decrease in blood TG concentration or blood cholesterol concentration in an individual administered with the anti-obesity agent, resulting in hyperlipidemia, arteriosclerosis, heart disease It is also useful for prevention and treatment. In addition, this anti-obesity agent leads to a reduction in the total cholesterol level in the blood by bringing about a significant decrease in the blood LDL cholesterol level while causing an increase in the blood HDL cholesterol level. Is extremely useful for.

  -The anti-obesity agent of this embodiment can be provided very easily and cheaply by containing the polar solvent extract which contains a hydrophilic component highly as an active ingredient. That is, the polar solvent extract can be produced by a very simple method of extracting a citrus fruit or its constituent components with a polar solvent.

<Extraction and purification of hydrophilic components from citrus fruits>
After extracting a polar solvent extract from the squeezed residue of lemon fruit, a hydrophilic component was obtained by performing a purification process. First, lemon fruit was squeezed with an FMC squeezing machine, and after the squeezed residue was collected, the squeezed residue was finely pulverized. Water as a polar solvent was added to 20 kg of the pulverized residue of the obtained juice residue, and the mixture was extracted with water for 30 minutes while stirring at room temperature (25 ° C.). The extract after stirring for 30 minutes was filtered to collect the filtrate, and then the filtrate was centrifuged at 9000 rpm for 20 minutes to obtain a supernatant (polar solvent extract).

  Subsequently, the supernatant was applied to a column packed with 2 L of an adsorption resin (Amberlite XAD-16 manufactured by Rohm and Haas), and the hydrophilic component in the supernatant was adsorbed on the adsorption resin. Next, 10 L of 18 ° C. water was passed through the column to wash the adsorbent resin, and then 10 L of a 30% by volume ethanol aqueous solution was passed through the column to obtain an eluate eluted with the ethanol aqueous solution. Finally, the obtained eluate was concentrated and dried to obtain a powdery hydrophilic component. The main substances contained in this hydrophilic component (lemon polyphenol) and their contents are shown in Table 1 below.

<Test on anti-obesity effect using obese mice>
Three groups of 8-week-old obese mice (female, C57BL6 / J manufactured by SLC, Japan) were prepared. Each group consisted of 6 mice, and was divided so that the average value of the weight of each group at the start of the test was the same between the groups. Each group of mice was fed on a normal diet for 5 days (CE-2 manufactured by CLEA Japan, Inc.) under the rearing conditions of a temperature of 23 ± 3 ° C., a humidity of 55 ± 10%, and a light cycle of 12 hours (lighted from 8:00:00 to 20:00). ) And deionized water and pre-bred. Next, while continuing the breeding conditions, the first control group was fed with the high carbohydrate diet shown in Table 2 below, the second control group was fed with the high fat diet of Table 2, and the test group (Example 2). Was fed with the high fat diet and hydrophilic components shown in Table 2 and started breeding. The hydrophilic component given to the test group was obtained in Example 1 above.

飼育開始から１週間毎に各群のマウスの体重を測定した。また、飼育開始から１４週後のマウスの体脂肪率をＤＥＸＡ法により測定した。また、飼育開始から１４週後のマウスを１２時間絶食させた後、エーテル麻酔下で腹部大動脈から血液を採取した。得られた各血液を３５００ｒｐｍで２０分間遠心分離することにより血清画分を得た。各血清画分について、血中ＴＧ濃度、血中総コレステロール濃度及び血中ＨＤＬコレステロール濃度をそれぞれ測定した。なお、血中ＴＧ濃度、血中総コレステロール濃度及び血中ＨＤＬコレステロール濃度は、それぞれトリグリセライドＧ−テスト、コレステロールＣII−テスト、ＨＤＬコレステロール−テスト（いずれも和光純薬工業社製）を用いて測定した。

The weight of each group of mice was measured every week from the start of breeding. In addition, the body fat percentage of mice 14 weeks after the start of breeding was measured by the DEXA method. In addition, the mice 14 weeks after the start of breeding were fasted for 12 hours, and then blood was collected from the abdominal aorta under ether anesthesia. Each blood obtained was centrifuged at 3500 rpm for 20 minutes to obtain a serum fraction. For each serum fraction, blood TG concentration, blood total cholesterol concentration, and blood HDL cholesterol concentration were measured. The blood TG concentration, blood total cholesterol concentration and blood HDL cholesterol concentration were measured using a triglyceride G-test, cholesterol CII-test, and HDL cholesterol-test (all manufactured by Wako Pure Chemical Industries, Ltd.), respectively. .

  The results are shown in FIGS. 1, 2 and 3 (a) to (c). In addition, the asterisk in FIG. 1 is statistically less than 5% of the second control group, the ** is less than 1%, and the *** is less than 0.1%. It has a significant difference. The *** mark in FIG. 2 indicates that there is a statistically significant difference with a risk rate of less than 0.1% relative to the second control group. In FIGS. 3A to 3C, the asterisk (*) indicates a risk rate of less than 5%, ** indicates a risk factor of less than 1%, and *** indicates a risk factor of less than 0.1%. It has a statistically significant difference.

  As shown in FIG. 1, the body weight of the second control group fed the high fat diet was significantly increased compared to the first control group fed the high carbohydrate diet. On the other hand, the increase in body weight was significantly suppressed in the mice of the test group to which the high fat diet + hydrophilic component was given compared to the mice of the second control group.

  As shown in FIG. 2, the body fat percentage was significantly increased in the second control group fed the high fat diet compared to the first control group fed the high carbohydrate diet. On the other hand, the increase in body fat percentage was significantly suppressed in the mice of the test group to which the high fat diet + hydrophilic component was given compared to the mice of the second control group.

  As shown in FIG. 3 (a), the TG concentration significantly increased in the second control group mice as compared to the first control group. In contrast, in the mice of the test group, the increase in TG concentration was significantly suppressed as compared with the second control group, and the concentration was lower than that of the first control group.

  As shown in FIG. 3 (b), the total cholesterol concentration was significantly increased in the mice of the second control group as compared with the first control group. In contrast, in the test group of mice, the total cholesterol concentration was not significantly increased compared to the first control group, and was lower than that of the second control group.

  As shown in FIG. 3 (c), the HDL cholesterol concentration was significantly increased in the second control group mice as compared to the first control group. On the other hand, in the mice of the test group, the HDL cholesterol concentration was significantly increased as compared with the first control group, and was higher than that of the second control group. Incidentally, HDL cholesterol, also called good cholesterol, is useful for preventing arteriosclerosis and the like because it has a function of recovering excess cholesterol from systemic tissues to the liver.

  When the results of FIGS. 3B and 3C are summarized, in the mice of the test group, the total cholesterol concentration is decreased and the HDL cholesterol concentration is increased compared to the second control group. That is, a selective increase in HDL cholesterol was observed in the test group of mice. Therefore, the citrus-derived hydrophilic component has the effect of suppressing body weight, suppressing the blood TG concentration, and increasing the blood HDL cholesterol concentration.

<Effect test on lipid metabolism enzyme gene expression>
Three groups of 8-week-old obese mice (C57BL6 / J manufactured by Japan SLC) were prepared. In the same manner as in Example 2, three groups of mice, a first control group fed with a high carbohydrate diet, a second control group fed with a high fat diet, and a test group fed with a high fat diet plus a hydrophilic component, Breeding started. After 10 weeks from the start of the breeding, the mice were fasted for 12 hours, and then the liver and calf muscle tissues were removed by surgery. MRNA was extracted from each of the extracted liver and muscle tissues, cDNA was prepared from each mRNA, and gene expression analysis was performed using a DNA microarray (manufactured by Affymetrix) using these cDNAs.

  The analyzed genes are ATP citrate lyase gene, acetyl CoA carboxylase gene, fatty acid synthase gene, and HMG-CoA reductase (Hydroxymethylglutaryl-CoA reductase). It is a gene. Regarding muscle tissue, they are a fatty acid CoA ligase (long chain 2) gene and an acyl CoA oxidase (ACO) gene. Table 3 shows the results of determining the magnification of the expression level of each gene in the test group relative to the expression level of each lipid metabolizing enzyme gene in the second control group.

その結果、高脂肪食＋親水性成分を与えた試験群では、肝臓において、脂肪酸合成系及びコレステロール合成系の遺伝子の発現がそれぞれ低下し、筋肉組織においては、脂肪酸燃焼系の遺伝子の発現が高まった。このことから、レモン由来の親水性成分は、遺伝子の発現レベルでも、脂肪酸及びコレステロールの合成を低下させるとともに、筋肉組織においては、脂肪酸のβ酸化を高める効果を有していることが確認された。

As a result, in the test group given a high fat diet + hydrophilic component, the expression of fatty acid synthesis system and cholesterol synthesis system genes decreased in the liver, and the expression of fatty acid combustion system genes increased in muscle tissue. It was. From this, it was confirmed that the hydrophilic component derived from lemon has the effect of reducing fatty acid and cholesterol synthesis at the gene expression level and increasing β-oxidation of fatty acid in muscle tissue. .

This embodiment can also be applied to the following technology.
In addition to the anti-obesity action, the active ingredient of the present embodiment is a blood TG concentration reducing action, a blood total cholesterol density reducing action, a blood LDL cholesterol density reducing action, and a blood HDL cholesterol density increasing action. have. Therefore, it is possible to provide a blood TG reducing agent, a blood cholesterol reducing agent, a blood LDL cholesterol reducing agent, and a blood HDL cholesterol increasing agent containing the active ingredient of this embodiment.

Further, the technical idea that can be grasped from the embodiment will be described below.
-The said body weight gain inhibitor characterized by having the effect | action which reduces a blood triglyceride density | concentration. The body weight gain inhibitor, which has an action of lowering blood low density lipoprotein cholesterol concentration. The weight gain inhibitor characterized by having an action of increasing blood high density lipoprotein concentration.

  -A blood low density lipoprotein cholesterol reducing agent comprising a hydrophilic component derived from citrus fruits as an active ingredient, wherein the active ingredient contains polyphenol. In this case, the blood low density lipoprotein cholesterol concentration can be reduced.

  A blood high density lipoprotein cholesterol increasing agent comprising a hydrophilic component derived from citrus fruits as an active ingredient, wherein the active ingredient contains polyphenol. In this case, the blood high density lipoprotein cholesterol concentration can be increased.

The graph which shows the measurement result of the body weight of the mouse | mouth of Example 2. FIG. The graph which shows the measurement result of the body fat percentage of the mouse | mouth of Example 2. FIG. (A)-(c) is a graph which shows the measurement result of the blood neutral fat density | concentration of the mouse | mouth of Example 2, the blood total cholesterol density | concentration, and the blood high density lipoprotein cholesterol density | concentration, respectively.

Claims (2)

The extract obtained from lemon juice residue with water, the hydrophilic component in the extract is adsorbed on the adsorbent resin, containing as active ingredient a component which is eluted with aqueous ethanol, by the aqueous ethanol The eluted component comprises eriocitrin, hesperidin, naryltin, and diosmin as main components, and is a weight gain inhibitor for the purpose of suppressing weight gain. About the extract obtained using water from lemon juice residue, the hydrophilic component in the extract is adsorbed to the adsorption resin, and the component eluted by the aqueous ethanol solution is contained as an active ingredient, thereby suppressing weight gain. In weight gain inhibitor for the purpose of
Body weight gain inhibitor you characterized as having the effect of increasing the expression of at least one gene selected from a fatty acid CoA ligase gene and acyl CoA oxidase gene.

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