JP2008156296A - Metabolic syndrome ameliorator - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a metabolic syndrome ameliorator helping to prevent and ameliorate metabolic syndrome using trihydroxyoctadecenoic acid (THODE) as a functional fatty acid alternative to trihydroxyoctadecadienoic acid (THODI), and to provide drinks and foods each containing the above metabolic syndrome ameliorator. <P>SOLUTION: The metabolic syndrome ameliorator comprises trihydroxyoctadecenoic acid (THODE) as an active ingredient. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an agent for improving metabolic syndrome.

  In recent years, lifestyle-related diseases such as hypertension, hyperlipidemia and diabetes have increased, and people's interest in health has increased. These lifestyle-related diseases are not independent of each other, but are related to each other due to obesity in which fat accumulates in the internal organs (visceral fat-type obesity). Recently, these lifestyle-related diseases have come to be called metabolic syndrome, and functional foods that are effective in preventing and improving metabolic syndrome have been actively developed.

  Trihydroxyoctadecadienoic acid (THODI) has been reported to have a preventive and ameliorating effect on metabolic syndrome, particularly a therapeutic effect on diabetes and a preventive effect on arteriosclerosis. (Non-patent Documents 1 and 2).

  On the other hand, trihydroxyoctadecenoic acid (THODE) is a fatty acid that is a causative substance of soybean bitterness and astringent taste of beer, but has an immunostimulatory action (Non-patent Document 3) and an antiallergic action during influenza virus infection. It is known to exert (Non-Patent Document 4).

Karageuzyan et al., Planta Medica, 1998, 64, p. 417-422 Orekhov et al., Phytomedicine, 1994, Volume 1, p. 123-126 Nagai et al., International Immunopharmacology, 2002, Volume 2, p. 1183-1193 Nagai et al., International Immunopharmacology, 2004, vol. 4, p. 1353-1365

  However, it is not known that trihydroxyoctadecenoic acid (THODE) has a preventive and ameliorating effect on metabolic syndrome.

  Therefore, an object of the present invention is to use trihydroxyoctadecenoic acid (THODE) as a functional fatty acid to replace trihydroxyoctadecadienoic acid (THODI), and to help prevent and improve metabolic syndrome. The objective of this invention is also providing the drink and foodstuff containing a metabolic syndrome improving agent.

  In order to achieve the above object, the present invention provides a metabolic syndrome improving agent comprising trihydroxyoctadecenoic acid (THODE) as an active ingredient.

  As a result of intensive studies on the application of fatty acids contained in plants to functional foods, the present inventors have found that trihydroxyoctadecenoic acid (THODE) has a preventive / improving effect on metabolic syndrome. Since trihydroxyoctadecenoic acid (THODE) is a fatty acid contained in soybeans, barley, and the like, safety to humans is ensured as compared with pharmaceuticals produced by artificial chemical synthesis. In addition, since trihydroxyoctadecenoic acid (THODE) has both an immunostimulatory action and an antiallergic action, it not only exerts a preventive and ameliorating effect on metabolic syndrome, but also enhances innate immunity and protects against viral infections such as colds. It can also contribute to prevention.

  The metabolic syndrome improving agent preferably has a blood pressure lowering effect.

  Hypertension is a major symptom of metabolic syndrome. If this symptom is left untreated, the heart and blood vessels are burdened, leading to more serious diseases such as arteriosclerosis, myocardial infarction, and cerebral infarction. The metabolic syndrome-improving agent can also be used in the treatment of hypertension as a blood pressure lowering agent, and can improve the pathology of metabolic syndrome.

  It is preferable that the metabolic syndrome improving agent has an action of lowering liver cholesterol concentration.

  An increase in liver cholesterol concentration is a major symptom of metabolic syndrome. If this condition is left untreated, it leads to an increase in cholesterol concentration in the blood, which is directly related to the development of hyperlipidemia and arteriosclerosis. The above-mentioned metabolic syndrome improving agent can be used as a hepatic cholesterol concentration reducing agent for preventing hyperlipidemia and arteriosclerosis, and can improve the pathology of metabolic syndrome.

  That is, the metabolic syndrome-improving agent of the present invention can also be used as a blood pressure lowering agent that suppresses an increase in blood pressure and / or a liver cholesterol concentration lowering agent that suppresses the accumulation of liver cholesterol.

  The above-described metabolic syndrome improving agent can be added to beverages and foods, and foods and drinks containing the metabolic syndrome improving agent have a blood pressure lowering effect and a liver cholesterol level reducing effect, and therefore can be used for dietary therapy of metabolic syndrome.

  Hereinafter, preferred embodiments for carrying out the present invention will be described.

  The metabolic syndrome improving agent of the present invention is characterized by containing trihydroxyoctadecenoic acid (THODE) as an active ingredient.

  In the present invention, “metabolic syndrome” is defined in “Definitions and diagnostic criteria of metabolic syndrome” by the Committee for Diagnostic Standards of Metabolic Syndrome (Journal of Japan Society for Internal Medicine, Vol. 94, No. 4, p. 794-809). Means what was done.

That is, in the case of Japanese, the waist circumference is ≥85 cm for men and ≥90 cm for women (in the case of visceral fat area, ≥100 cm ² is equivalent for both men and women). And
(1) Lipoprotein abnormality: hypertriglyceridemia (triglyceride level ≧ 150 mg / dL) and / or low HDL cholesterolemia (HDL cholesterol level <40 mg / dL),
(2) High blood pressure: systolic blood pressure (≧ 130 mmHg) and / or diastolic blood pressure (≧ 85 mmHg),
(3) Hyperglycemia (fasting hyperglycemia ≧ 110 mg / dL)
Are included in the diagnostic criteria.

  According to the above diagnostic criteria, when metabolic syndrome is diagnosed, it means that lifestyle-related diseases such as hypertension, hyperlipidemia and diabetes have occurred, and serious diseases such as arteriosclerosis, myocardial infarction and cerebral infarction have occurred. It can be said that the disease is likely to be caused.

  Trihydroxyoctadecenoic acid (THODE) is a commercially available compound, but can be produced by chemical synthesis by itself.

  Trihydroxyoctadecenoic acid (THODE) is a suspension process in which a raw material plant is pulverized to obtain a slurry liquid, and an enzyme treatment in which β-glucanase is added to the slurry liquid obtained in the suspension process to perform an enzyme treatment. Residue, removing the residue from the enzyme-treated slurry liquid in the enzyme treatment step, obtaining a supernatant, adding n-hexane to the supernatant obtained in the removal step, stirring the aqueous layer The hexane extraction process to be recovered, ether added to the aqueous layer recovered in the hexane extraction process and stirred, the ether extraction process to recover the ether layer, and the drying process to dry the ether layer recovered in the ether extraction process And can be purified from the raw material plant by a purification method comprising:

  According to the above purification method, trihydroxyoctadecenoic acid (THODE) contained in a plant can be efficiently extracted without using a purification means such as column chromatography. That is, the above purification method can be used to purify trihydroxyoctadecenoic acid (THODE) from a raw plant at the factory level, and the obtained trihydroxyoctadecenoic acid (THODE) is used for the production of the metabolic syndrome improving agent. it can.

  The raw material plant corresponds to a plant containing trihydroxyoctadecenoic acid (THODE), and in particular, cereals, specifically rice, wheat, wheat, corn, straw, and cane can be exemplified.

  In the suspension step, first, the raw material plant is pulverized by adding an aqueous solvent so that trihydroxyoctadecenoic acid (THODE) present in the raw material plant is easily extracted into the aqueous solvent. This crushing may be performed by homogenizing the raw plant using, for example, a homogenizer, a mortar or the like. The amount of water added is preferably equal to or greater than the mass of the raw material plant, and more preferably 1.5 times the amount.

  In the enzyme treatment step, the enzyme treatment is performed by adding β-glucanase to the slurry and stirring. The temperature of the enzyme treatment is preferably 50 ° C. or higher, more preferably 60 ° C. The pH during the enzyme reaction is preferably pH 3 or more and pH 5 or less, more preferably pH 4. Any β-glucanase may be used as long as it can degrade β-glucan, which is a polysaccharide component of the cell wall of the raw material plant, and is easily available from the market. For example, when using Genencor's β-glucanase (Optimash BG; 10300 CMCU / g, 1.05-1.15 g / mL), 5 mL of β-glucanase may be added to 1 kg of raw material plant. . The reaction time is preferably 15 hours or more and 24 hours or less.

  In the removal step, plant residues that are solids are removed from the slurry after the enzyme treatment. For example, large residues can be removed by passing the slurry liquid through a 10-20 mesh sieve, and small residues can be removed as precipitates by subsequently centrifuging the slurry liquid.

  In the hexane extraction step, 1/10 equivalent of n-hexane may be added to the supernatant collected in the removal step and stirred so that the aqueous layer and the organic layer are well mixed. As a result, components soluble in hexane can be removed.

  In the ether extraction step, 1/10 to 1/5 equivalent of diethyl ether is added to the aqueous layer recovered in the hexane extraction step and stirred so that the aqueous layer and the organic layer are well mixed. As a result, trihydroxyoctadecenoic acid (THODE) soluble in diethyl ether can be extracted into the organic layer.

  In the drying step, moisture in the organic layer is removed. For example, anhydrous sodium sulfate or anhydrous magnesium sulfate can be used as a desiccant. Moreover, it can concentrate and remove a water | moisture content by carrying out pressure reduction processing.

  Moreover, the metabolic syndrome improving agent of the present invention is characterized by having a blood pressure lowering action.

  The blood pressure lowering agent refers to a drug having an action of lowering the increased blood pressure to a normal level. Hypertension is a major symptom of metabolic syndrome, so if the above-mentioned metabolic syndrome improving agent is administered to a patient diagnosed with metabolic syndrome, secondary symptoms of metabolic syndrome such as arteriosclerosis, myocardial infarction, cerebral infarction, etc. Can prevent progress to. For example, in a diet therapy for a hypertensive patient or a patient diagnosed with metabolic syndrome, an increase in blood pressure can be suppressed by ingesting the above-described metabolic syndrome improving agent together with a normal meal.

  Moreover, the metabolic syndrome-improving agent of the present invention is characterized by having an action of lowering liver cholesterol concentration.

  The liver cholesterol concentration reducing agent refers to a drug having an action of suppressing excessive accumulation of cholesterol in the liver. Increased liver cholesterol level is a major symptom of metabolic syndrome, so if the above-mentioned metabolic syndrome improving agent is administered to a patient diagnosed with metabolic syndrome, secondary symptoms of metabolic syndrome, arteriosclerosis, myocardial infarction It is possible to prevent progression to cerebral infarction and the like. For example, in the diet therapy for a patient diagnosed with metabolic syndrome, excessive intake of cholesterol in the liver can be suppressed by ingesting the metabolic syndrome-improving agent together with a normal meal.

  The metabolic syndrome-improving agent of the present invention is effective in improving the pathology of metabolic syndrome in the metabolic syndrome diet, and can be used by adding to beverages or foods.

  Beverages and foods containing the above-described metabolic syndrome improving agent may contain additives commonly used in the art. Examples of this additive include apple fiber, soybean fiber, meat extract, black vinegar extract, gelatin, corn starch, honey, animal and vegetable oils and fats, monosaccharides such as glucose, disaccharides such as sucrose, fructose and mannitol, dextrose and starch, etc. Polysaccharides, sugar alcohols such as erythritol, xylitol, and sorbitol, and vitamins such as vitamin C. These additives may be used alone or in combination.

  In addition, the above-mentioned metabolic syndrome improving agent may be added to foods for specified health foods, special nutritional foods, nutritional supplements, health foods, functional foods, foods for the sick, alcoholic beverages and soft drinks. it can.

  EXAMPLES Hereinafter, although this invention is demonstrated more concretely based on an Example and a comparative example, this invention is not limited to a following example at all.

Example 1 Effect of Trihydroxyoctadecenoic Acid (THODE) on Blood Pressure and Liver Cholesterol Concentration in Spontaneously Obese Model Mice Experimental Material 1) Test Substance Trihydroxyoctadecenoic acid (THODE) was purchased from Larodan Fine Chemicals (Sweden) and stored in a cool and dark place until used for experiments.

2) Animals used Ob / ob mice (Slc: C57BL / 6J Ham ob / ob, 5 weeks old, male) were purchased from Japan SLC Co., Ltd., and were abnormal after pre-breeding including a one-week quarantine period. Animals that were not recognized were selected for use in the following experiments. Each individual was identified by marking the animal number with an ear punch and a felt pen on the tail, and the containment cage was attached with a card with a group number and an animal number.

  The ob / ob mouse is a spontaneous obesity model mouse characterized by almost no ability to synthesize leptin, which is one of anti-obesity factors. For this reason, there is no suppression of the feeding center, obesity associated with overeating, hyperglycemia and hyperinsulinemia, which are accompanied by abnormal lipid metabolism and abnormal glucose metabolism. This is very similar to the pathogenesis of human obesity, and is considered to be the most suitable model animal for food screening tests that are expected to have an effect on metabolic syndrome and lifestyle-related diseases.

  The breeding room was set to have a temperature of 22 ± 3 ° C., a humidity of 50 ± 20%, a lighting time of 12 hours (8:00:00 to 20:00), and a ventilation rate of 13-17 times / hour throughout the preliminary breeding and the experiment period. . Ob / ob mice were housed in a polysulfone housing cage (outer diameter 265W × 427D × 204H mm) 4-5 each, and the housing cages were mounted on a stainless steel movable rack (1790W × 470D × 1650H mm). The feed is solid feed (AIN-93G; Oriental Yeast Co., Ltd.) fed from a stainless steel solid feed feeder, so that the water can be freely supplied from a polysulfone water feeder (made of a stainless steel pipe) containing tap water. did.

3) Administration test to ob / ob mice Ob / ob mice that had completed the preliminary breeding period were divided into 8 groups per group using systolic blood pressure as an index. Table 1 shows the dose of trihydroxyoctadecenoic acid (THODE) administered orally to each group of mice.

  In administration group 1, 18 μg of trihydroxyoctadecenoic acid (THODE) was dissolved in 0.5 mL of purified water, and in administration group 2, 180 μg of trihydroxyoctadecenoic acid (THODE) was purified in 0.5 mL. In the control group 1, 0.5 mL of purified water was orally administered once a day for 28 days.

  From the day after the start of administration to the 29th day, body weight, food intake and water intake were measured over time. On days 7, 14, 21, and 28 after the start of administration, the systolic blood pressure 1 hour after administration was tailed. Measurement was performed by the cuff method. On the 29th day after the start of administration, the animals were fasted for 3 hours and then anesthetized with pentobarbital (50 mg / kg, ip), followed by laparotomy, and the liver was removed. Liver cholesterol concentration and liver weight were measured from the extracted liver.

4) Statistical processing The average value and standard error in each group were calculated from the obtained numerical values. The significant difference between the control group 1 and the administration groups 1 and 2 is tested for equivariance by the Bartlett method, further one-way analysis of variance is performed in the case of equal variance, and if significant, the mean value is compared by the Dunnett method. Went. In the case of unequal variance, the Kruskal-Wallis H test was performed, and when significant, the average rank was compared by the Dunnett method. Significance levels were 10%, 5% and 1%.

2. Experimental results 1) Effect of administration of trihydroxyoctadecenoic acid (THODE) on body weight, food intake and water intake FIG. 1 shows the change in the body weight of mice from the day after the start of oral administration to the 29th day. It is the result of examining with time.

  As a result, an increase in body weight with an increase in age was confirmed in all animals of each group. The body weight fluctuation of the mice in the administration groups 1 and 2 is almost the same as the body weight fluctuation of the mice in the control group 1, and the amount of trihydroxyoctadecenoic acid (THODE) administered orally in the administration groups 1 and 2 is Did not affect the body weight fluctuation.

  FIG. 2 shows changes in the amount of food intake per day from the day after the day when oral administration was started to day 29, and FIG. 3 shows the daily change from the day after the day when oral administration was started to day 29. It is the result of having investigated the fluctuation | variation of the water intake of the mouse | mouth of this time.

  As a result, the daily food intake and water intake during the administration period tended to decrease over time in any group, but the food intake and water intake in the mice in the administration groups 1 and 2 The fluctuations were almost the same as the fluctuations in food intake and water consumption in the mice of the control group 1, and no significant difference was observed between the administration groups 1 and 2 and the control group 1.

  Based on the above results, the mice in administration groups 1 and 2 grew almost the same as the mice in control group 1, and the administration of trihydroxyoctadecenoic acid (THODE) showed severe toxicity to the mice. It was suggested that there was no problem in the safety of trihydroxyoctadecenoic acid (THODE).

2) Effect of trihydroxyoctadecenoic acid (THODE) on blood pressure and heart rate FIG. 4 shows the results of examining changes in systolic blood pressure of mice during the administration period of trihydroxyoctadecenoic acid (THODE) over time. is there.

  As a result, the systolic blood pressure of the control group 1 mice increased with increasing age. On the other hand, in administration group 1, a decrease in systolic blood pressure was observed from day 21 after the start of administration, and a significant difference (p <0.01) compared to control group 1 on days 21 and 28 after the start of administration. Was recognized. In administration group 2, a decrease in systolic blood pressure was observed from day 14 after the start of administration, and a significant difference (p <0. 01) was observed. The heart rate of each group of mice was also measured simultaneously with the measurement of systolic blood pressure, but no fluctuation in the heart rate was confirmed in any group, and trihydroxyoctadecenoic acid (THODE) was added to the mouse heart rate. There was no effect.

3) Effect of trihydroxyoctadecenoic acid (THODE) on liver cholesterol concentration FIG. 5 shows the results of examining the liver cholesterol concentration of mice at the end of administration of trihydroxyoctadecenoic acid (THODE).

  As a result, in the administration group 1, although the liver cholesterol concentration tended to slightly decrease, there was no significant difference compared to the control group 1, but in the administration group 2, the liver cholesterol concentration tended to decrease significantly. Was observed, and a significant difference was observed compared with the control group 1 (p <0.05). The liver weights of the mice in each group were also measured at the same time. In each group, the liver weight was almost the same, and trihydroxyoctadecenoic acid (THODE) did not affect the liver weight of the mice. .

(Example 2) Effect of trihydroxyoctadecenoic acid (THODE) extracted from beer lees on blood pressure and liver cholesterol concentration in spontaneously obese model mice Experimental material 1) Test substance Trihydroxyoctadecenoic acid (THODE) is obtained by extracting beer cake (hereinafter referred to as malt feed) produced after squeezing wort from malt used for brewing beer as follows. The composition was used in the experiment as a test substance.

  First, 2.0 kg (water content 80%) of malt feed is placed in a 5 L Erlenmeyer flask, suspended by adding 3.0 L of distilled water, and the malt flask is immersed in a water bath set at 60 ° C. The feed suspension was kept warm. Then, while stirring the suspension with a stirrer, the pH was adjusted to 4.0 by adding 6N hydrochloric acid, and 10 mL of β-glucanase (Optimash BG; Genencor) was added and reacted with stirring. At that time, in order to prevent evaporation of hot water in the water bath, a plastic ball was floated over the surface of the water, and further covered with aluminum foil. In this state, stirring is continued overnight to allow reaction, and then the suspension is passed through a 10-20 mesh sieve to remove larger plant residues (such as husks) and further, 2,000 rotations for 12 minutes. Centrifugation removed small plant residues as precipitates. The supernatant thus obtained was transferred to a separating funnel, to which 1/10 equivalent of n-hexane was added and mixed to perform hexane extraction, and the aqueous layer was recovered. The aqueous layer from which the hexane-soluble component was removed was transferred to a new separatory funnel, to which 1/10 to 1/5 equivalent of diethyl ether was added and mixed to perform ether extraction, and the ether layer was recovered. The ether layer thus obtained was dried using anhydrous sodium sulfate and then concentrated to dryness under reduced pressure. As a result, a composition containing 12.5% (mass%) of trihydroxyoctadecenoic acid (THODE) (hereinafter, trihydroxyoctadecenoic acid (THODE) composition) was obtained.

2) Animals used As in Example 1, ob / ob mice (Slc: C57BL / 6J Ham ob / ob, 5 weeks old, male) were purchased from Japan SLC Co., Ltd. and abnormalities were observed after preliminary breeding. Animals that could not be selected were used for the following experiments. The breeding room, breeding conditions, food, etc. were the same as in Example 1.

3) Administration test to ob / ob mice Ob / ob mice that had completed the preliminary breeding period were divided into 8 groups per group using systolic blood pressure as an index. Table 2 shows the dose of trihydroxyoctadecenoic acid (THODE) composition orally administered to each group of mice.

  In administration group 3, 1000 μg of trihydroxyoctadecenoic acid (THODE) composition was dissolved in 0.5 mL of 1% Tween 80 aqueous solution, and in control group 2, 0.5 mL of 1% Tween 80 aqueous solution was added. Repeated oral administration once a day for 28 days.

  On days 7, 14, 21, and 28 after the start of administration, systolic blood pressure 1 hour after administration was measured by the tail cuff method. On day 29 after administration, pentobarbital was fasted for 3 hours. Anesthesia was performed at (50 mg / kg, ip) and the abdomen was opened, and the liver was removed. Liver cholesterol concentration and liver weight were measured from the extracted liver.

4) Statistical processing An average value and a standard error in each group were calculated from the obtained numerical values, and a significant difference test was performed according to the method described in Example 1.

2. Experimental Results 1) Effect of Trihydroxyoctadecenoic Acid (THODE) Composition on Blood Pressure and Heart Rate FIG. 6 shows changes in the systolic blood pressure of mice during the administration period of the trihydroxyoctadecenoic acid (THODE) composition over time. It is the result of examining it.

  As a result, the systolic blood pressure of the control group 2 mice increased with increasing age. On the other hand, in the administration group 3, after 14 days after the start of administration, a significant decrease in blood pressure was observed compared to the control group 2 (p <0.01). The heart rate of each group of mice was also measured at the same time as the measurement of systolic blood pressure, but no change in the heart rate was confirmed in any group, and the trihydroxyoctadecenoic acid (THODE) composition was used in the mouse heart rate. The number was not affected.

2) Effect of trihydroxyoctadecenoic acid (THODE) composition on liver cholesterol concentration FIG. 7 shows the results of examining the liver cholesterol concentration in mice at the end of administration of the trihydroxyoctadecenoic acid (THODE) composition.

  As a result, in the administration group 3, the liver cholesterol concentration tended to decrease significantly, and a significant difference was observed compared to the control group 2 (p <0.01). The liver weights of the mice in each group were also measured at the same time. In each group, the liver weights were almost the same, and the trihydroxyoctadecenoic acid (THODE) composition had an effect on the liver weights of the mice. There wasn't.

It is the graph which investigated the weight change of the ob / ob mouse | mouth from the day after the day of starting oral administration to the 29th day over time. It is the graph which investigated the fluctuation | variation of the food intake of the ob / ob mouse | mouth per day from the day after the day which started oral administration to the 29th day. It is the graph which investigated the fluctuation | variation of the water intake of the ob / ob mouse | mouth per day from the day following the day of the start of oral administration to the 29th day. It is the graph which investigated the fluctuation | variation of the systolic blood pressure of the ob / ob mouse | mouth during the administration period of a trihydroxy octadecenoic acid (THODE) with time. It is the graph which investigated the liver cholesterol level of the ob / ob mouse | mouth at the time of completion | finish of administration of a trihydroxyoctadecenoic acid (THODE). It is the graph which investigated the fluctuation | variation of the systolic blood pressure of the ob / ob mouse | mouth during the administration period of a trihydroxyoctadecenoic acid (THODE) composition with time. It is the graph which investigated the liver cholesterol level of the ob / ob mouse | mouth at the time of completion | finish of administration of a trihydroxy octadecenoic acid (THODE) composition.

Claims (5)

  A metabolic syndrome improving agent comprising trihydroxyoctadecenoic acid (THODE) as an active ingredient.   The metabolic syndrome improving agent according to claim 1, which has a blood pressure lowering effect.   The metabolic syndrome-improving agent according to claim 1, which has an action of lowering liver cholesterol concentration.   The drink containing the metabolic syndrome improving agent according to any one of claims 1 to 3.   The foodstuff containing the metabolic syndrome improving agent as described in any one of Claims 1-3.

Images