JP5606005B2 - Metabolic syndrome improvement or prevention agent - Google Patents

Description

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

  In recent years, metabolic syndrome (a state in which lifestyle-related diseases such as hypertension, hyperlipidemia, and diabetes have developed due to visceral fat accumulation) increases the risk of developing arteriosclerotic diseases (myocardial infarction, cerebral infarction, etc.) Widely recognized as a thing.

  As an agent for improving or preventing metabolic syndrome, for example, an agent containing a dietary fiber aggregate having a maximum content of water-soluble β-glucan as an active ingredient is known (see Patent Document 1).

International Publication No. 2007/077929 Pamphlet

  By the way, as a metabolic syndrome improving or preventing agent, it is desirable to have a high safety for a living body and can be ingested daily and continuously. However, with respect to such metabolic syndrome improving or preventing agents, there are still no sufficient options for satisfying the diverse demands of consumers.

  Then, this invention makes it a subject to provide the novel metabolic syndrome improvement or prevention agent which has high safety | security with respect to a biological body, and can be ingested daily and continuously.

  When the present inventors administer low molecular weight β-1,3-1,4-glucan to mice, the present inventors administer high molecular weight β-1,3-1,4-glucan contained in the barley extract. As compared with the above, the present inventors have found that the symptoms of metabolic syndrome including visceral fat accumulation are strongly suppressed, and the present invention has been completed.

  That is, the present invention provides a metabolic syndrome improving or preventing agent containing β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da as an active ingredient.

  The agent for improving or preventing metabolic syndrome of the present invention makes it possible to suppress visceral fat accumulation and suppress fat cell hypertrophy. And, through such an action, visceral fat type obesity or metabolic syndrome can be improved (treated, reduced) and prevented. Here, “inhibition of accumulation” of visceral fat refers to suppressing increase of visceral fat or reducing visceral fat.

  The metabolic syndrome improving or preventing agent of the present invention can also improve liver function and suppress accumulation of liver triglycerides. Through such actions, it is possible to suppress (treat, reduce, prevent) liver damage (fatty liver, hepatitis, liver fibrosis, cirrhosis, liver cancer, etc.). Here, “inhibition of accumulation” of liver triglyceride refers to inhibiting increase of liver triglyceride or reducing liver triglyceride. Whether liver function has been improved can be determined by, for example, whether plasma AST (aspartate aminotransferase) has decreased.

  Since the metabolic syndrome improving or preventing agent of the present invention has a visceral fat accumulation inhibitory action and an adipocyte hypertrophy inhibitory action, it can also be used as a visceral fat accumulation inhibitor or an adipocyte hypertrophy inhibitor.

  Since the metabolic syndrome improving or preventing agent of the present invention also has a liver function improving effect, a liver triglyceride accumulation inhibitory effect and a liver injury inhibitory effect, it is used as a liver function improving agent, a liver triglyceride accumulation inhibitor or a liver disorder inhibitor. You can also.

  β-1,3-1,4-glucan is abundantly contained in gramineous plants (especially barley and oats), and safety for living bodies has been established. Therefore, the metabolic syndrome improving or preventing agent of the present invention is highly safe to the living body and can be ingested daily and continuously, as a component of pharmaceuticals, foods and drinks, food and drink additives, feeds, feed additives and the like. Suitable for use.

  In addition, β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da is higher in water solubility and lower in viscosity of the solution than those having a high molecular weight. Also in this respect, the metabolic syndrome improving or preventing agent of the present invention is suitable for use as a component of pharmaceuticals, foods and drinks, food and drink additives, feeds, feed additives and the like.

  ADVANTAGE OF THE INVENTION According to this invention, the safety | security with respect to a biological body is high, and the novel metabolic syndrome improvement or prevention agent which can be ingested daily and continuously is provided. Moreover, the pharmaceutical, food / beverage products, food / beverage product additive, feed, feed additive, etc. containing such a metabolic syndrome improvement or prevention agent are provided.

It is a molecular weight distribution curve of β-1,3-1,4-glucan in β-glucan powder. It is a graph which shows the mesentery fat cell size of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the accessory testicle periphery fat cell size of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the abdominal wall fat cell size of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the amount of total lipid in the feces of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the mesentery fat weight of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the weight of fat around the epididymis of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the back abdominal wall fat weight of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the intraperitoneal total fat weight of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the mesentery fat cell size of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the plasma AST of the mouse | mouth which administered (beta) -1,3-1,4-glucan. It is a graph which shows the amount of liver triglycerides of the mouse | mouth which administered (beta) -1,3-1,4-glucan.

  Hereinafter, embodiments of the present invention will be described in detail.

  The metabolic syndrome improving or preventing agent of the present invention contains β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da as an active ingredient.

In the present invention, “metabolic syndrome” refers to a state in which visceral fat is accumulated and any lifestyle-related diseases such as hypertension, hyperlipidemia, and diabetes develop. For example, in the case of Japanese, it satisfies the requirement of “waist circumference: male ≧ 85 cm, female ≧ 90 cm (both men and women correspond to visceral fat area ≧ 100 cm ² )”, and the following (1) to (3) Metabolic syndrome can be diagnosed if at least two of these requirements are met (see Journal of the Japan Society for Internal Medicine, 94 (4), 794-809, 2005).
(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 blood glucose ≧ 110 mg / dL

  In the present invention, the β-1,3-1,4-glucan may be a β-glucan having a β-1,3-glucoside bond and a β-1,4-glucoside bond. Or it may be derived from microorganisms (bacteria, basidiomycetes, etc.). As a natural product for obtaining β-1,3-1,4-glucan, for example, a grass family (barley, oats, rye, And wheat are preferred, and barley and oats are particularly preferred. In addition, for example, when obtained from a grass family plant, seeds are suitable (any of whole grains, endosperm, cocoons, etc.).

  The β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da, for example, is obtained by pulverizing barley seeds and treating the pulverized product with α-amylase to obtain a high molecular weight β-1,3-1. , 4-glucan (weight average molecular weight: 700,000 to 1,200,000 Da) can be obtained and further treated with β-glucanase. Here, the α-amylase treatment of the barley seed pulverized product can be performed, for example, by stirring the pulverized product and α-amylase in water at 80 to 95 ° C. for 40 to 60 minutes. After the α-amylase treatment, solid-liquid separation is performed, and the liquid layer is precipitated with alcohol (methanol, ethanol, etc.) and concentrated to obtain high molecular weight β-1,3-1,4-glucan. The β-glucanase treatment of high molecular weight β-1,3-1,4-glucan is performed, for example, in water at 50 to 60 ° C. for 5 to 30 minutes, for high molecular weight β-1,3-1,4-glucan and β -It can be carried out by stirring with glucanase. After concentration with β-glucanase, β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da can be obtained. In addition, as for the β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da, if it is commercially available, it may be used.

  In the present invention, the weight average molecular weight of β-1,3-1,4-glucan may be 50,000 to 500,000 Da, for example, preferably 50,000 to 300,000 Da, and 50,000 to 200,000 Da. More preferred is 50,000 to 150,000 Da.

  The weight average molecular weight of β-1,3-1,4-glucan can be measured by a known method (for example, gel permeation chromatography (GPC), gel filtration chromatography (GFC)). Moreover, the calibration curve used for determination of a weight average molecular weight can be created, for example using the standard product of a pullulan.

  The metabolic syndrome improving or preventing agent of the present invention may be in any form of solid (for example, powder obtained by freeze-drying), liquid (water-soluble or fat-soluble solution or suspension), paste, etc. Any dosage form such as powders, granules, tablets, capsules, solutions, suspensions, emulsions, ointments, plasters and the like may be used. Moreover, the metabolic syndrome improving or preventing agent of the present invention may be composed of β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da.

  The above-mentioned various preparations are composed of β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da and pharmaceutically acceptable additives (excipients, binders, lubricants, disintegrations). Agents, emulsifiers, surfactants, bases, solubilizers, suspending agents, and the like).

  For example, the excipient includes lactose, sucrose, starch, dextrin and the like. Examples of the binder include polyvinyl alcohol, gum arabic, tragacanth, gelatin, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone and the like. Examples of the lubricant include magnesium stearate, calcium stearate, talc and the like. Examples of the disintegrant include crystalline cellulose, agar, gelatin, calcium carbonate, sodium bicarbonate, dextrin and the like. Examples of the emulsifier or surfactant include Tween 60, Tween 80, Span 80, and glyceryl monostearate. Examples of the base include cetostearyl alcohol, lanolin, polyethylene glycol, rice bran oil, fish oil (DHA, EPA, etc.), olive oil and the like. Examples of the solubilizer include polyethylene glycol, propylene glycol, sodium carbonate, sodium citrate, Tween 80 and the like. Examples of the suspending agent include polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, hydroxymethyl cellulose, sodium alginate and the like in addition to the above-described surfactant.

  The metabolic syndrome improving or preventing agent of the present invention can be used as a component of medicine, food and drink (beverage, food), food and drink additive, feed, feed additive and the like. For example, examples of the beverage include water, soft drinks, fruit juice beverages, milk beverages, alcoholic beverages, sports drinks, and nutritional drinks. Examples of foods include breads, noodles, rice, tofu, dairy products, soy sauce, miso, and confectionery. The metabolic syndrome improving or preventing agent of the present invention can also be used as a component for food for specified health use, food for special use, dietary supplement, health food, functional food, food for sick people, and the like.

  The content ratio of the metabolic syndrome improving or preventing agent (β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da) in beverages, foods, feeds, etc. is, for example, 0.01 to 20% by mass. Is preferable, 0.01-10 mass% is more preferable, 0.01-5 mass% is still more preferable.

  Beverages, foods, feeds and the like may further contain additives usually used in the field. Examples of such additives include bitters, flavorings, apple fiber, soybean fiber, meat extract, black vinegar extract, gelatin, corn starch, honey, animal and vegetable oils and fats; proteins such as gluten; beans such as soybeans and peas; glucose Monosaccharides such as fructose; disaccharides such as sucrose; polysaccharides such as dextrose and starch; sugar alcohols such as erythritol, xylitol, sorbitol and mannitol; vitamins such as vitamin C; minerals such as zinc, copper and magnesium Functional materials such as CoQ10, α-lipoic acid, carnitine, capsaicin; These additives can be used alone or in combination of two or more.

  The metabolic syndrome improving or preventing agent of the present invention may be ingested by humans or non-human mammals. The intake amount and the intake method can be appropriately determined according to the state, age, etc. of the individual. As a suitable intake method, for example, oral intake can be mentioned.

  Hereinafter, based on an Example, this invention is demonstrated more concretely. However, the present invention is not limited to the following examples.

[Preparation and analysis of high molecular weight β-glucan powder]
6 kg of barley (CDC Fibar) seed pulverized product and 300 g of heat-resistant α-amylase (Chrystase YC15, Yamato Kasei) were put into about 60 L of hot water (about 40-50 ° C.) in a stainless steel extraction tank (200 L), The mixture was stirred at 95 ° C. for 60 minutes. Thereafter, 10 L of the mixed solution was dispensed into a plastic tank and centrifuged at 20 ° C. and 8000 rpm for 20 minutes. About 45 L of the obtained translucent, high-viscosity supernatant was put into a drum can lift (250 L), about 140 L of methanol was added, and after sufficient stirring, the mixture was allowed to stand for 1 hour.

  After standing, the mixed solution was poured onto a stainless steel sieve (φ400) to separate a solid. After washing with 20 L of methanol and removing the methanol with an explosion-proof evaporator, the solid was freeze-dried on a stainless steel pad. The freeze-dried solid was intermittently pulverized for 2 minutes with a lab miller (IFM-150, Iwatani Corporation) to obtain a high molecular weight β-glucan powder.

  The viscosity of a 0.2% aqueous solution of high molecular weight β-glucan powder was 3.49 mPa · s.

  The molecular weight distribution of β-1,3-1,4-glucan in the high molecular weight β-glucan powder was as shown in FIG. The weight average molecular weight of β-1,3-1,4-glucan in the high molecular weight β-glucan powder was about 900,000 Da. The molecular weight distribution and the weight average molecular weight were measured by gel permeation chromatography (GPC). The GPC conditions are as follows. Two pumps (pumps A and B) were used, and eluents A and B were allowed to flow through pumps A and B, respectively. As a sample for analysis, a filtrate obtained by filtering a solution of 10 mg of high molecular weight β-glucan powder in 10 mL of water through a 0.45 μm filter was used. The molecular weight distribution and the weight average molecular weight were determined based on a calibration curve prepared using a pullulan standard (Shodex).

GPC conditions:
・ Oven temperature: 40 ℃
Column: Shodex OHPak SB-805HQ (excluding molecular weight 4 million)
+ Shodex OHPak SB-804HQ (excluding 1 million molecular weight)
+ Shodex OHPak SB-803HQ (excluding molecular weight 100,000)
-Mixing coil: Stainless steel tube with an inner diameter of 0.5 mm and an empty volume of 0.5 mL-Eluent A: Ultrapure water Flow rate: 1 mL / min-Eluent B: Calcoflow solution Flow rate: 0.5 mL / min-HPLC device: Shimadzu LC-10 Series
System controller: SCL-10Avp
Pump: LC-10ATvp
Oven: CTO-10ACvp
Autosampler: SIL-10ADvp
Detector: RID-10A, RF-10AxL
・ Analysis software: GPC analysis software for Class-VP, Class-VP ・ Detector: differential refractive index (RI) detector (temperature: 40 ° C.); fluorescence (FL) detector (excitation wavelength: 360 nm; fluorescence wavelength: 420 nm) )
・ Injection volume: 100 μL
・ Analysis time: 40 minutes

  The component composition of the high molecular weight β-glucan powder was as shown in Table 1. In the table, the unit of each component amount is mass%. In addition, the amount of β-1,3-1,4-glucan is measured by FL analysis using calcoflow.

[Preparation and analysis of low molecular weight β-glucan powder]
10 L of water was put into a stainless steel plant micro extractor (20 L), heated to 80 ° C., and then 300 g of high molecular weight β-glucan powder was added. After stirring, 10 L of water and 300 g of high molecular weight β-glucan powder were added, and the mixture was stirred at 80 ° C. for 1 hour. After cooling to 62 ° C. and adding 300 mg of β-glucanase (Shin Nippon Chemical Co., Ltd.), the temperature was raised again to 80 ° C. and held for 10 minutes. Thereafter, the mixed solution was freeze-dried on a stainless steel pad, and the obtained solid was intermittently pulverized with a lab miller (IFM-150, Iwatani Corporation) for 2 minutes to obtain a low molecular weight β-glucan powder.

  The viscosity of a 0.2% aqueous solution of low molecular weight β-glucan powder was 1.20 mPa · s.

  The molecular weight distribution of β-1,3-1,4-glucan in the low molecular weight β-glucan powder was as shown in FIG. Further, the weight average molecular weight of β-1,3-1,4-glucan in the low molecular weight β-glucan powder was about 100,000 Da. The molecular weight distribution and the weight average molecular weight were measured in the same manner as in the case of the high molecular weight β-glucan powder.

[Test Example 1]
(Mouse grouping)
24 mice (4 weeks old, male, C57BL / 6J mouse, Charles River Japan) in good general condition, and 3 groups (cellulose 8) in each group so that the initial body weight (average) does not vary between groups Group, high molecular weight β-glucan group, low molecular weight β-glucan group).

(Preparation of test feed)
The test feed to be administered to each group of mice was prepared based on the powdered feed AIN93G so as to obtain the composition shown in Table 2. In the table, the unit of each component amount is g / kg feed.

(Test feed administration)
After one week of acclimatization, each group of mice was allowed to freely ingest a predetermined test feed and tap water for 8 weeks. Throughout the acclimatization period and the subsequent test feed administration period, the mice had a temperature of 22 ± 3 ° C., a relative humidity of 55 ± 20%, a ventilation rate of 12 times / hour, and a light / dark time of 12 hours (light period: 8 hours to 20 hours). They were reared in groups. During the acclimation breeding period, AIN93G was used as it was as feed.

(Measurement of fat cell size and total lipid content in feces)
After completion of the test feed administration period, each group of mice was subjected to cardiac blood sampling and dissection under ether anesthesia, and the mesenteric fat cell size, the epididymal fat cell size, the abdominal wall fat cell size and the total fecal lipid amount were determined. It was measured.

(result)
The results (mean ± standard deviation) are shown in Tables 3 and 4 and FIGS. 2 to 4 are graphs showing the mesenteric fat cell size, the accessory testicle peripheral fat cell size, and the retroabdominal wall fat cell size of each group of mice, respectively. FIG. 5 is a graph showing the total amount of lipid in feces of each group of mice.

  As is apparent from Table 3 and FIGS. -It showed a low value compared to the glucan group, and particularly a significantly low value compared to the cellulose group. Further, as is apparent from Table 4 and FIG. 5, the total amount of fecal lipids in the low molecular weight β-glucan group is higher than that in the cellulose group and the high molecular weight β-glucan group, and particularly in comparison with the cellulose group. The value was remarkably high.

[Test Example 2]
(Mouse grouping)
30 mice with good general condition (4 weeks old, male, C57BL / 6J mouse, Charles River Japan), 3 groups (10 cellulose each) so that the initial weight (average) does not vary between groups Group, high molecular weight β-glucan group, low molecular weight β-glucan group).

(Preparation of test feed)
The test feed to be administered to each group of mice was prepared in the same manner as in Test Example 1.

(Test feed administration)
After one week of acclimatization, each group of mice was allowed to freely ingest a predetermined test feed and tap water for 8 weeks. Throughout the acclimatization period and the subsequent test feed administration period, the mice had a temperature of 22 ± 3 ° C., a relative humidity of 55 ± 20%, a ventilation rate of 12 times / hour, and a light / dark time of 12 hours (light period: 8 hours to 20 hours). Individually reared under conditions. During the acclimation breeding period, AIN93G was used as it was as feed.

(Measurement of fat weight, adipocyte size, plasma AST, liver triglyceride content)
After completion of the test feed administration period, each group of mice was subjected to cardiac blood sampling and dissection under ether anesthesia, mesenteric fat weight, epididymal fat weight, retroabdominal wall fat weight, mesenteric fat cell size, plasma AST And the amount of liver triglyceride was measured. Plasma AST is generally used as an indicator of liver function.

  The amount of liver triglyceride was measured by the Folch method. Specifically, first, the liver of the mouse was excised and freeze-dried. After 0.2 g of freeze-dried liver was extracted with 20 mL of chloroform / methanol (2: 1 (volume ratio)) overnight, centrifugation (3000 rpm, 10 minutes) was performed, and the supernatant was collected. Next, 10 mL of chloroform / methanol (2: 1 (volume ratio)) was added to the precipitate obtained by centrifugation, and the mixture was stirred and further centrifuged (3000 rpm, 10 minutes) to recover the supernatant. All the obtained supernatants were combined and filtered through a filter paper (No. 2), and the filtrate was made up to 50 mL with chloroform / methanol (2: 1 (volume ratio)) to obtain a liver lipid extract. 5 mL of the extract was collected and washed with a blank for Folch washing [chloroform / methanol / water (3:28:47 (volume ratio))], and the solvent was evaporated to dryness at 60 ° C. in a nitrogen stream. This was dissolved in 0.5 mL of isopropanol to obtain a test solution. The amount of triglyceride in the test solution was measured using Triglyceride E-Test Wako (Wako Pure Chemical Industries).

(result)
The results (mean ± standard deviation) are shown in Tables 5 to 11 and FIGS. FIGS. 6 to 9 are graphs showing mesenteric fat weight, epididymal fat weight, retroabdominal wall fat weight and intraperitoneal total fat weight of each group of mice, respectively. 6-9, the graph of (a) shows the fat weight per individual, and the graph of (b) shows the fat weight per 100 g of body weight. FIG. 10 is a graph showing the mesenteric adipocyte size of each group of mice. FIG. 11 is a graph showing plasma AST of mice in each group. FIG. 12 is a graph showing the amount of liver triglyceride in each group of mice. The total fat weight in the abdominal cavity corresponds to the sum of the mesenteric fat weight, the peripheral testicular fat weight, and the retroabdominal wall fat weight.

  As is apparent from Tables 5-11 and FIGS. 6-12, mesenteric fat weight, peripheral weight of the epididymis, retroabdominal wall fat weight, intraperitoneal total fat weight, mesenteric fat cell size, plasma AST and liver triglycerides All of the amounts were low in the low molecular weight β-glucan group as compared to the cellulose group and the high molecular weight β-glucan group, and particularly significantly lower than the cellulose group.

  According to the above examples, β-1,3-1,4-glucan having a weight average molecular weight of 50,000 to 500,000 Da is a visceral fat accumulation inhibitory action, an adipocyte hypertrophy inhibitory action, a liver function improving action, and a liver triglyceride accumulation. It has been confirmed that it has an inhibitory action and is useful as a component for improving or preventing metabolic syndrome.

Claims (1)

A metabolic syndrome relieving or preventing agent containing beta-1,3-1,4-glucan having a weight-average molecular weight from 50,000 to 150,000 Da as an active ingredient,
An agent for improving or preventing metabolic syndrome , which is used as a liver function improving agent , wherein the liver function is a liver function using plasma AST (aspartate aminotransferase) as an index .

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