JP5503125B2 - Visceral fat accumulation inhibitor - Google Patents

Description

  The present invention relates to a flavonoid composition containing rutin, isoquercitrin, and α-glucosylated products thereof, visceral fat accumulation-suppressing action of this composition, and foods and beverages, pharmaceuticals and the like containing such a composition. .

  The convenience of being able to eat whatever you like as you go along with the progress of food culture has led to overeating and unbalanced eating, and the lack of exercise and stressed society have spurred this and created the term “metabolic syndrome”. In general, metabolic syndrome refers to accumulation of visceral fat (waist is 85 cm or more for men, 90 cm or more for women), blood lipids (neutral fat 150 mg / dL or less or HDL cholesterol less than 40 mg / dL), hungry It is defined as a case where two or more items are selected from blood glucose (100 mg / dL or more) and blood pressure (blood pressure of contraction 130 mmHg or more). On the other hand, there are various supplements, health appliances, and the like with anti-metabolic syndrome. In particular, products that reduce weight, which are most easily understood in appearance, and products that focus on reducing visceral fat that induces various diseases are noticeable.

  In the case of visceral fat type obesity, it is said that lipid metabolism and sugar metabolism are decreased by adipocyte enlargement and secretion of adiponectin, which is a useful cytokine, decreases, and secretion of TNF-α, an inflammatory substance, increases. Yes. As a result, blood lipid increases, cholesterol and the like are deposited on the blood vessel wall, and a rod-like plaque is formed. Plaque inhibits the function of vascular endothelial cells and gradually hardens to induce arteriosclerosis.

Among the foods and beverages and pharmaceuticals containing flavonoids, the following inventions have been proposed so far as having relevance to blood glucose and blood lipids.
JP 2004-059522 A (Patent Document 1) discloses an enzyme-treated rutin and a persistent rutin preparation containing rutin and an oral composition (food, medicine) containing the same, cancer, Applications for prevention and treatment of diseases such as arteriosclerosis and ischemic heart disease have been suggested.

  However, although the example of Patent Document 1 shows that when the oral composition is ingested, the concentration of quercetin conjugate is maintained in blood for a long time, an effect on a specific disease or symptom, for example, No data showing a specific action such as suppression of visceral fat adhesion is disclosed.

  JP 2008-007452 A (Patent Document 2) discloses a pancreatic β-cell protective agent containing a flavonoid compound (such as α-glucosyl rutin) or tannic acid as an active ingredient, a food or drink containing the same, and a pharmaceutical product. Such pancreatic β-cell protective agents are suggested to be used to improve pancreatic fatigue, abnormal carbohydrate metabolism, abnormal lipid metabolism, abnormal protein metabolism, hyperlipidemia, hypertension, metabolic syndrome, etc. Has been. And, when the pancreatic β-cell protective agent is brought into contact with the pancreatic β-cell line, an effect of recovering the decreased insulin secretion activity appears (Example 1), and 2% fermented soybean extract (high content of daidzein and genistein) Ingestion of glucose and lipid metabolism, such as suppression of blood glucose level increase, increase in blood triglyceride concentration, increase in blood total cholesterol concentration, increase in subcutaneous fat weight (Example) 3) etc. are shown.

JP 2002-524480 A (Patent Document 3) discloses the use of a plant extract containing a bioflavonoid (such as rutin) for lowering blood sugar in mammals, foods and beverages containing these, drugs, and the like. Has been. In the Examples, it is shown that the administration of rutin or the like has the effect of reducing the blood sugar level.

  However, Patent Documents 2 and 3 disclose excellent effects of suppressing visceral fat accumulation in compositions containing rutin and / or isoquercitrin and α-glucosylrutin and / or α-glucosylisoquercitrin. It is not specifically disclosed to have

Regarding actions such as lipid metabolism improvement, the amount of cholesterol and neutral fat in normal blood and organs is usually measured, and as a means of screening, increase or decrease of adiponectin and TNF-α is confirmed from the expression of adipocyte genes It is common to evaluate the presence or absence of an effect by a technique such as. However, it cannot always be said that a compound or the like evaluated to have an action of improving lipid metabolism by these techniques actually has an action of suppressing the accumulation of visceral fat, and an action of suppressing the visceral fat accumulation is often not observed.
JP 2004-059522 A JP 2008-007452 A JP-T-2002-524480

  An object of the present invention is to provide a visceral fat accumulation inhibitor that is useful for the prevention and treatment of metabolic syndrome, obesity, and the like and can be blended in foods and drinks, pharmaceuticals, and the like.

  As a result of diligent research, the present inventors have found that rutin and / or isoquercitrin (hereinafter abbreviated as “flavonoid”) and α-glucosylrutin and / or α-glucosylisoquercitrin (hereinafter “flavonoid derivatives”). And a flavonoid composition containing the abundant visceral fat (in particular, abdominal fat), the present invention has been completed.

  That is, the visceral fat accumulation inhibitor of the present invention is characterized by comprising the above flavonoid composition having a visceral fat accumulation inhibiting action. The weight ratio of the flavonoid / flavonoid derivative (corresponding to rutin) in the flavonoid composition is preferably 40/60 to 10/90.

  Such a visceral fat inhibitor can be contained in, for example, food and drink, pharmaceuticals, feed or pet food. It is preferable to prepare such foods and drinks, pharmaceuticals and the like so that the flavonoid composition is orally ingested in an amount based on 0.01 to 3 g / kg mouse body weight / day. In particular, such pharmaceuticals can be used for the prevention or treatment of metabolic syndrome or obesity.

  The visceral fat accumulation inhibitor of the present invention exhibits an excellent visceral fat accumulation inhibitory effect particularly when used in combination with a high fat diet, and shows a tendency to reduce visceral fat even when used in combination with a normal diet. (Obesity) and effective prevention and treatment of metabolic syndrome.

− Components of visceral fat accumulation inhibitor −
-Rutin Rutin is represented by the formula (I), with β-lutinose (6-O
-Α-L-rhamnosyl-β-D-glucose) is a quercetin glycoside having a structure bound thereto.

The method for obtaining and preparing rutin used in the present invention is not particularly limited, and it is possible to use what is manufactured and sold as a reagent, or from raw materials such as Enju flower buds or buckwheat young leaves. You may use what was extracted and refine | purified.

Isoquercitrin Isoquercitrin is a compound represented by formula (II) in which β-D-glucose is bonded to the hydroxyl group at the 7-position of quercetin, that is, a compound in which the rhamnose residue in the rutinose unit of rutin is cleaved. It is.

  The method for obtaining and preparing isoquercitrin to be used in the present invention is not particularly limited, and it is possible to use an enzyme that is manufactured and sold as a reagent or the like, or an enzyme having rhamnosidase activity in rutin (for example, hesperidinase, naringinase). Those prepared by acting may be used.

Α-Glucosylrutin α-glucosylrutin has one or more (about 2 to 20) glucose bound to the glucose residue in the rutinose unit of rutin represented by formula (III) by α1 → 4 bond. A compound. Of these, those bound with only one glucose are also called “monoglucosyl rutin”.

α-Glucosylrutin is a glycosyltransferase (cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19), etc.) in the presence of α-glucosyl sugar compounds (cyclodextrin, partially degraded starch, etc.) and glucose in rutin. It can be produced by the action of an enzyme having a function of adding. The α-glucosylrutin obtained by such an enzyme treatment is usually a mixture comprising different numbers of bound glucose, that is, monoglucosylrutin and other α-glucosylrutins.

In addition, monoglucosylrutin causes α-glucosylrutin to act on α-glucosylrutin by causing an enzyme having glucoamylase activity, such as glucoamylase (EC 3.2.1.3), that cleaves α-1,4-glucoside bonds in glucose units, as described above. It can be produced by cleaving leaving only one glucose bound to rutin.

-Α-glucosyl isoquercitrin α-glucosyl isoquercitrin is represented by formula (IV), and one or more (about 2 to 20) glucose is added to the glucose residue of isoquercitrin by α1 → 4 bond. It is a bound compound. Among these, one having only one glucose bonded is also called “monoglucosyl isoquercitrin”.

As in the case of α-glucosyl rutin described above, α-glucosyl isoquercitrin is converted to glycosyltransferase (cyclodextrin) in the presence of an α-glucosyl sugar compound (cyclodextrin, partially decomposed starch, etc.). It can be produced by allowing glucanotransferase (CGTase, EC 2.4.1.19) or the like to act on an enzyme having the function of adding glucose to isoquercitrin.

-Mixing ratio The visceral fat accumulation inhibitor (flavonoid composition) of the present invention is rutin and / or isoquercitrin (hereinafter abbreviated as "flavonoid") in consideration of the solubility and bioabsorbability of the composition. , Α-glucosylrutin and / or α-glucosylisoquercitrin (hereinafter abbreviated as “flavonoid derivative”), the weight ratio of flavonoid / flavonoid derivative (corresponding to rutin) is 40/60 to 10/90. It is preferable to contain by quantity.

  Here, the “equivalent amount of rutin” is a value obtained according to a stoichiometric calculation. For example, in HPLC, the peak area of a rutin standard solution having a known concentration and each component (α-glucosyl of an aqueous solution of a flavonoid composition). It is calculated | required by comparing with the peak area of rutin, isoquercitrin, etc.).

− Method for producing visceral fat accumulation inhibitor −
The visceral fat accumulation inhibitor of the present invention, that is, a composition comprising predetermined flavonoids and flavonoid derivatives such as rutin, isoquercitrin, α-glucosylrutin, α-glucosylisoquercitrin basically comprises each component (preferably By mixing at a predetermined ratio as described above.

  The method for that is not particularly limited, and purified products of the above components may be prepared separately and then mixed, but rutin is used as a starting material, and the preparation methods of the respective components as described above are combined. It is preferable to prepare a composition containing the above components by carrying out a series of enzyme treatments, and to carry out further treatment as necessary.

  For example, under appropriate enzyme treatment conditions, cyclodextrin glucanotransferase is allowed to act on rutin to prepare a mixture of rutin (unreacted) and α-glucosylrutin, and then glucoamylase and rhamnosidase are allowed to act on this mixture. Then, by producing monoglucosylrutin from α-glucosylrutin and isoquercitrin from rutin, a composition containing monoglucosylrutin and isoquercitrin as main components can be obtained.

  Moreover, you may perform another process as needed after such an enzyme treatment process or in the middle. For example, filtration treatment to remove precipitates, concentration treatment that does not produce precipitates, desalting treatment using ion exchange resin, purification treatment to remove other impurities, and further from these liquids Examples thereof include a drying or lyophilization treatment for preparing a solid material.

  The preparation method for each component used in the present invention and the production method of a composition containing a plurality of components by a series of enzyme treatments are known. For example, the reaction conditions (temperature, pH, enzyme, etc.) of the enzyme treatment and the product For details of the purification method and the like, Japanese Patent No. 2926411 (for α-glucosyl rutin and the like), Japanese Patent No. 3155466 (for isoquercitrin, monoglucosyl rutin and the like), Japanese Patent Application Laid-Open No. 2003-261593 (α-glucosyl). References such as isoquercitrin) can be referred to.

  The composition obtained by the production method as described above is generally produced and sold as “enzyme-treated rutin”, and it is convenient and suitable to use such a product in the present invention. For example, the product “αG rutin PS” manufactured by Toyo Seika Co., Ltd. is a composition containing 75% by weight of α-glucosyl rutin and 25% by weight of isoquercitrin. Similarly, a product “αG rutin P” manufactured by Toyo Seika Co., Ltd. is a composition containing 75% by weight of α-glucosyl rutin and 25% by weight of rutin.

− Food and drink, pharmaceuticals, feed, pet food −
The visceral fat accumulation inhibitor of the present invention is formed into an appropriate form such as granules, granules, tablets, capsules, pastes, etc. after adding appropriate auxiliary agents and various nutritional components as necessary. Can be ingested.

  The visceral fat accumulation inhibitor of the present invention is blended in food and drink (including health functional foods and other so-called health foods and supplements), pharmaceuticals (including quasi drugs), feed, pet food, and the like. Can also be used. In particular, the above pharmaceutical products can be used for the prevention or treatment of metabolic syndrome or obesity.

The types of foods and drinks that can contain the visceral fat accumulation inhibitor of the present invention are not particularly limited. For example, fruit drinks, oolong tea, green tea, black tea, cocoa, vegetable juice, green juice, soy milk, milk Examples include beverages such as beverages, lactic acid beverages, near water, sports drinks, and nutritional drinks, Western confectionery such as jelly, pudding, and yogurt, Japanese confectionery, seasonings, fish meat, processed fish products, and processed livestock products. Such foods and drinks should be mixed, kneaded, dissolved, soaked, sprayed, sprayed, applied, etc. with a liquid or solid visceral fat accumulation inhibitor (flavonoid composition) using general means for food production. Can be manufactured.

  The pharmaceutical preparation containing the visceral fat accumulation inhibitor of the present invention is preferably an oral preparation such as a liquid, syrup, drink, tablet, capsule, powder, fine granule, etc. A dosage form suitable for an administration method such as internal administration or injection can also be used. If necessary, it can be produced by using conventional means such as non-toxic pharmaceutical carriers, stabilizers, binders, and other conventional additives.

  The effective dose of such pharmaceuticals can be appropriately determined according to the patient's age, weight, symptoms, lifestyle, administration route of the drug, administration schedule, formulation, strength of the inhibitory activity of the material, etc. . For example, oral administration of the visceral fat accumulation inhibitor of the present invention in an amount based on 0.01 to 3 g / kg mouse body weight / day is effective in suppressing visceral fat accumulation and is suitable for human administration. The amount can also be adjusted based on the above range. In addition, also when ingesting the visceral fat accumulation inhibitor of this invention as food and drink etc., said amount can be used as a standard. Moreover, it is preferable to take the medicine containing the visceral fat accumulation inhibitor after a meal, especially after each meal.

(1) Animal feed As a high fat diet, feed “MF” (manufactured by Oriental Yeast Co., Ltd.) containing 30% by weight lard, and feed containing 5.3% by weight normal lard as a control diet (control diet) “MF” (same as above) was prepared. Furthermore, a feed containing 0.5% by weight of “αG rutin PS” (manufactured by Toyo Seika Co., Ltd.) was prepared for both these diets. In order to prevent lipid oxidation, the high fat diet was individually packaged with an oxygen scavenger in an amount of 100 g, and all feeds were stored at 4 ° C. until use.

(2) Feed intake After purchase of 4-week-old male C57BL / 6 mice (Japan SLC Co., Ltd.), the control MF feed and ion-exchanged water were bred freely in the breeding gauge for 5 days for 10 days. (Acclimation period). The breeding room was maintained at a humidity of 50-60% and a room temperature of 23 ± 1 ° C., and was lit from 8:00 to 20:00. After the acclimatization period, they were grouped as follows.

a) Control group: intake of control food (MF feed containing lard 5.3% by weight) b) Control + 0.5% rutin group ... control food intake containing 0.5% αG rutin PS c) High fat group: lard 30% D) High-fat diet + 0.5% rutin group ... 0.5% αG rutin PS-containing high-fat diet The experiment was conducted with 10 animals per group and 5 animals / cage. Food consumption was measured twice a week using “Roden Cafe” (manufactured by Oriental Yeast Co., Ltd.), and body weight was measured once a week for 12 weeks. This experiment was conducted according to the guidelines for animal experiments at Shizuoka Prefectural University.

(3) Dissection and sampling After 6 hours of fasting, dissection was performed. Laparotomy was performed after ether anesthesia, and blood was collected. The blood was subjected to heparin treatment and then centrifuged (2,000 g, 5 minutes) to obtain plasma. The liver, adrenal gland, and kidney were weighed, and the liver and kidney were rapidly frozen using liquid nitrogen and stored at ultra-low temperature. In addition, the fat around the testis and other visceral fat were fractionated, and their weights were measured.

(4) Results and Discussion Mice in the high fat diet group showed significant weight gain compared to the control group (Table 1, FIG. 1). In particular, the increase was significant after 7 weeks from the start of feeding. Since the calorie intake was almost the same under the breeding conditions (FIG. 2), this effect is not due to the difference in the amount of intake energy. The high fat diet + 0.5% rutin group showed significant weight gain compared to the control group, but the body weight was significantly suppressed (P <0.05) compared to the high fat diet group (Table 1).

  In addition, the ingestion of the high-fat diet for 12 weeks significantly increased the weight of fat around the testis and visceral fat (P <0.05) than that of the control diet (Table 1). This result suggests that one of the causes of weight gain associated with high fat diet intake is due to changes in these fat weights.

  However, in the high fat diet + 0.5% rutin group, the visceral fat weight of the mice was significantly suppressed (P <0.05) compared to the high fat diet group. In addition, the control + 0.5% rutin group also showed a tendency that the visceral fat weight of the mice was lower than that of the control group.

  In addition, mice in the high fat diet group showed a marked increase in blood leptin concentration compared to the control group (FIG. 3). This result is presumed that the leptin concentration increased in proportion to the increase in visceral fat. However, in the high fat diet + 0.5% rutin group, the blood leptin concentration was significantly suppressed (P <0.05) compared to the high fat diet group.

  The blood insulin concentration was significantly increased in the high fat diet group compared to the control group, and was significantly decreased in the high fat diet + 0.5% rutin group as compared to the high fat diet group (FIG. 4). On the other hand, the blood sugar level of the high fat diet group was the same level as that of the control group (FIG. 5). This is presumed that the blood sugar level is suppressed by the increased insulin. In the future, if the intake of a high fat diet continues, the mechanism will be disrupted, and there is a high possibility that blood sugar level will rise and diabetes will be induced. However, even if a high fat diet is ingested, the blood insulin level and blood glucose level can be maintained at the control level by ingesting 0.5% αG rutin PS at the same time.

  From the above, it is speculated that daily intake of rutin suppresses body weight gain and increase in visceral fat (intraperitoneal fat) weight, and is one factor in the anti-obesity effect particularly when a large amount of fat is consumed.

The graph showing the amount of weight gain at the time of ingestion of the alpha G rutin addition food of the mouse | mouth in an Example. The graph showing transition of the cumulative intake amount at the time of ingestion of the αG rutin-added food of mice in the examples. The graph showing the blood leptin density | concentration at the time of a (alpha) G rutin addition food intake of the mouse | mouth in an Example. The graph showing the insulin density | concentration in the plasma at the time of a (alpha) G rutin addition food intake of the mouse | mouth in an Example. The graph showing the blood glucose level at the time of ingesting the alpha G rutin addition food of the mouse | mouth in an Example.

Claims (4)

A visceral fat accumulation inhibitor comprising a flavonoid composition containing isoquercitrin and α-glucosyl rutin and having a visceral fat accumulation inhibitory action. The weight ratio of isoquercitrin / alpha-glucosyl rutin in the flavonoid composition (rutin equivalent) is 40 / 60-10 / 90, visceral fat accumulation inhibitor according to claim 1. A medicinal product comprising the visceral fat accumulation inhibitor according to claim 1 or 2 for preventing or treating metabolic syndrome or obesity . Characterized in that the flavonoid composition is orally ingested in an amount based on 0.01 to 3 g / kg mouse body weight / day, medicament according to claim 3.