JP2007195510A - Calorie-restricting food - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a calorie-restricting food capable of preventing, treating and/or improving diabetes mellitus and obesity. <P>SOLUTION: The calorie-restricting food comprises an inhibitor having inhibiting activity against α-glucosidase as an active ingredient. The inhibitor includes an antagonism inhibitor and a non-antagonism inhibitor. The antagonism inhibitor is preferably Salacia extract obtained by extracting root and/or stem of Salacia reticulata of the family Hippocrateaceae or the family Celastraceae with a solvent. The non-antagonism inhibitor is preferably Coleus extract obtained by extracting a root of Coleus froskohlii which is a plant of the family Labiatae with a solvent. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a calorie-restricted food containing as an active ingredient an inhibitor having an inhibitory activity against α-glucosidase.

  Obesity is a risk factor for many diseases such as ischemic heart disease, hypertension, and diabetes, and obesity is responsible for 30 to 60% of patients with hypertension, hyperlipidemia, and diabetes Is estimated. Furthermore, obesity in Japan has increased 2 to 4 times over the past 30 years, which is a major factor threatening the health of the people. In the future, lifestyle-related diseases are expected to increase as the population ages, and obesity prevention is becoming an important issue to be overcome not only from the health aspect but also from the social and economic aspects.

  Over 90% of obesity is caused by an excess of energy intake relative to energy consumed, which is called simple obesity. To prevent this, there are (1) suppression of intake energy and (2) promotion of energy consumption.

For (1), methods such as dietary restriction, appetite suppressant administration, suppression of absorption by adsorption of lipids, etc. in the digestive tract, formation of satiety due to indigestible dietary fiber, and administration of digestive enzyme inhibitors, etc. is there.
(2) includes exercise therapy, administration of energy metabolism promoters, and the like.

  In particular, inhibiting digestive absorption of carbohydrates in the digestive tract by inhibiting a disaccharide-degrading enzyme such as α-glucosidase is very effective in preventing excessive intake of calories. Glucobuy (registered trademark, general name: Acarbose, Bayer Yakuhin Kogyo), Basin (registered trademark, Takeda Yakuhin Kogyo), etc. are also used as anti-diabetic drugs focusing on calorie restriction. These inhibitors of α-glucosidase are considered to be α-glucosidase substrate analogs (analogs) because of their structures.

  Therefore, the mode of action is considered to be antagonistic inhibition that inhibits binding of the original saccharide, which is a disaccharide, to the enzyme by binding the analog to the binding site of the enzyme with the substrate. On the other hand, by binding to a site other than the binding site with the enzyme substrate, for example, the three-dimensional structure of the enzyme is changed, and the binding force with the enzyme substrate is lost, or the active expression site of the enzyme is inactivated. Substances having an action are called non-competitive inhibitors.

  As a disaccharide-degrading enzyme inhibitor obtained from a natural plant, for example, there is a solvent extract of Saratinareticata of Dechinmuraceae or Nishigiidae (see Patent Document 1). A substance obtained from this salatiareticurata is a competitive inhibitor and has α-glucosidase inhibitory activity. In addition, there is an extract of Coleus Forskori obtained by extracting Coleus Forskori, which is a Labiatae plant native to India, with water or alcohol (see Patent Document 2). The substance obtained from this Coleus forskori is a non-competitive inhibitor and has α-glucosidase inhibitory activity.

However, inhibition of the enzyme by competitive inhibitors depends on the concentration of the substrate. For this reason, when the concentration of the original substrate is increased, the substrate pushes out the inhibitor and binds to the enzyme, and as a result, the sugar decomposition reaction may proceed.
Inhibition of an enzyme by a non-competitive inhibitor does not depend on the concentration of the substrate, but in general, since the reaction with the enzyme occurs at 1: 1, the quantity of each of the enzyme and the inhibitor is the inhibition efficiency. Greatly affects. Therefore, the efficacy of non-competitive inhibitors is greatly influenced by the amount of enzyme secreted. On the other hand, the amount of secreted enzyme varies from individual to individual, and even if it is the same individual, the range of fluctuation is large. For example, the amount of secreted pancreatic juice is known to vary from 700 to 1500 ml daily (non-patent). Reference 1). Therefore, the secreted enzyme may vary greatly depending on, for example, eating habits, etc., and the efficacy of non-competitive inhibitors may also vary greatly.
Therefore, it is difficult for conventional antagonistic inhibitors or noncompetitive inhibitors to sufficiently suppress the intake energy according to different internal environments, and it has not always been effective in preventing diabetes and obesity.

JP 2004-323420 A JP 2001-206893 A Mitsuyasu Shiraso, “Examination of pancreatic exocrine function by pancreatic drainage method”, Journal of the Japanese Society of Gastroenterology, 1973, Vol. 70, No. 7, p. 658-672

  This invention is made | formed in view of this conventional problem, Comprising: It aims at providing the calorie restriction food which can prevent, treat, and / or improve diabetes and obesity.

The present invention is a calorie-restricted food containing an inhibitory substance having an inhibitory activity against α-glucosidase as an active ingredient,
The inhibitory substance is a calorie-restricted food containing a competitive inhibitory substance and a non-antagonistic inhibitory substance (Claim 1).

The calorie-restricted food of the present invention contains the competitive inhibitor and the non-competitive inhibitor as the inhibitor.
Therefore, the calorie-restricted food can sufficiently inhibit α-glucosidase even in environments with various substrate concentrations and enzyme concentrations due to the interaction between the antagonistic inhibitor and the non-antagonistic inhibitor. Therefore, the calorie-restricted food hardly inhibits the activity of α-glucosidase in the human body and is absorbed by being decomposed into monosaccharides and hardly depending on the type of individual and the body environment of the individual. Can be suppressed. Moreover, the said calorie restriction food can produce a feeling of fullness with respect to the person who ingested this calorie restriction food.

  As described above, the calorie-restricted food can inhibit the activity of α-glucosidase and can generate a feeling of fullness. Therefore, the intake energy can be suppressed, and diabetes and obesity can be prevented, treated and / or improved.

Next, a preferred embodiment of the present invention will be described.
The calorie-restricted food contains the competitive inhibitor and the non-competitive inhibitor as the inhibitor for α-glucosidase.
The antagonistic inhibitor is also called a competitive inhibitor, and is a substance that inhibits the binding between the substrate and the enzyme by binding to or near the binding site with the substrate in the enzyme (α-glucosidase) while competing with the substrate. Say. For example, there is a substrate analog (analog) that binds to the α-glucosidase substrate binding site but is not metabolized.

  In addition, the non-competitive inhibitor is also called an anti-competitive inhibitor, and by binding to a site other than the binding site with the substrate in the enzyme (α-glucosidase), for example, changes in the three-dimensional structure of the enzyme, A substance having an action of eliminating the binding force of an enzyme with a substrate or inactivating an enzyme active expression site.

The antagonistic inhibitor preferably contains salacinol and / or kotalanol (claim 2).
In this case, the calorie restricted food can exhibit more excellent α-glucosidase inhibitory activity.
The structural formula of salacinol is shown in the following “Chemical Formula 1”, and the structural formula of kotalanol is shown in the following “Chemical Formula 2”.

Further, the antagonistic inhibitor is preferably a Salacia extract obtained by extracting a root and / or stem of Salacia reticulata of the Dechinmuraceae or Nigiriidae with a solvent (Claim 3).
The non-antagonistic inhibitor is preferably a Coleus extract obtained by extracting a root of Coleus forskori, a Labiatae plant, with a solvent (claim 4).
In these cases, the calorie-restricted food contains a natural ingredient as an active ingredient and can exhibit higher safety for the human body and can exhibit excellent α-glucosidase inhibitory activity.

  As said solvent for extracting the said Salacia extract and the said Coleus extract, water, alcohol, these mixtures etc. can be used, for example. Specifically, methanol, ethanol, isopropanol or the like can be used as the alcohol.

Example 1
Next, examples of the calorie restricted food of the present invention will be described.
In this example, a calorie-restricted food is prepared and its function is evaluated.
The calorie-restricted food of this example contains an inhibitory substance having an inhibitory activity against α-glucosidase as an active ingredient. In addition, the calorie-restricted food contains a competitive inhibitor and a non-competitive inhibitor as inhibitors. In this example, in particular, a calorie-restricted food containing Salacia extract as a competitive inhibitor and a Coleus extract as a non-competitive inhibitor is prepared.

First, a Salacia extract is prepared as follows.
Specifically, first, 0.5 L of ethanol was added to 100 g of a ground product of Salacia reticulata root, and extraction was performed at room temperature for 3 hours (reflux). Subsequently, it isolate | separated into the extract (henceforth the extract A) and the extraction residue by filtration.
Next, 0.5 L of water was added to this extraction residue, extraction was performed at room temperature for 3 hours, and then an extraction liquid (hereinafter referred to as extraction liquid B) was obtained by filtration.

  Next, the extract A and the extract B were concentrated and dried under reduced pressure to obtain the extract A and the extract B, respectively. These extract A and extract B were mixed, pulverized and sieved to obtain a Salacia extract (sample C1).

Next, a Coleus extract is prepared as follows.
That is, first, 500 mL of a mixed solvent of ethanol and water (ethanol concentration: 50% by volume) was added to 100 g of powdered Coleus forskori roots, and extraction was performed at a temperature of 50 to 60 ° C. for 3 hours. Subsequently, it isolate | separated into the extract and the extraction residue by filtration.

  Next, this Coleus extract was concentrated and dried under reduced pressure, pulverized, and sieved to obtain a Coleus extract (sample C2).

  Next, 50 g of the Salacia extract (sample C1) obtained as described above and 50 g of the Coleus extract (sample C2) were mixed to prepare a calorie-restricted food of this example. This is designated as Sample E.

Next, the inhibitory activity on the α-glucosidase of the calorie restricted food (Sample E) was examined by the method of Vogel & Vogel.
Specifically, first, an enzyme-substrate reaction is performed. That is, a mixed solution obtained by mixing 250 μL of pH 7.0 phosphate buffer, sample E, and 50 μL of enzyme solution was incubated at a temperature of 37 ° C. for 30 minutes. As the enzyme solution, an invertase (concentration 4 units / mL) manufactured by Wako Pure Chemical Industries, Ltd., which is a kind of α-glucosidase and has a decomposition activity for sucrose (sucrose), was used 10 times.
Next, 500 μL of the substrate solution was added to this mixed solution, and incubated at a temperature of 37 ° C. for 20 minutes. As a substrate solution, 37 mM sucrose was used. The reaction was then stopped by immersing in boiling water for 2 minutes and then cooled. Next, 50 μL of this reaction solution was added to 250 μL of a glucose reagent, mixed, and incubated at room temperature for 10 minutes. This is designated as test sample X1.

  Next, the above-mentioned enzyme-substrate reaction was performed without using sample E. That is, a mixed solution obtained by mixing 250 μL of pH 7.0 phosphate buffer and 50 μL of enzyme solution (0.4 unit / mL invertase) was incubated at a temperature of 37 ° C. for 30 minutes. Next, in the same manner as in the case of the test sample X1, the substrate solution (37 mM sucrose) was added to the mixture and incubated, and the reaction was stopped in boiling water and cooled, and then the reaction solution was added to the glucose reagent. Mix and incubate. This is designated as test sample Y1.

Next, the absorbance at a wavelength of 510 nm was measured for the test sample X1 and the test sample Y1. When the absorbance of the test sample X1 is X1, and the absorbance of the test sample Y1 is Y1,% inhibitory activity (I) was calculated based on the following formula (1). In this example, the above-described enzyme-substrate reaction was performed while changing the concentration of sample E, and the% inhibitory activity of sample E on sucrose-degrading enzyme was calculated. The result is shown in FIG.
I = (Y1-X1) / Y1 × 100 (1)

  In addition, as a positive control, instead of sample E, an acarbose that is already known to have an inhibitory activity against sucrose-degrading enzyme was used to carry out an enzyme-substrate reaction, and the inhibitory activity of acarbose on sucrose-degrading enzyme (% inhibitory activity) Was measured in the same manner as in Sample E described above. In this example, the above-described enzyme-substrate reaction was carried out while changing the concentration of acarbose, and the% inhibitory activity of acarbose on sucrose-degrading enzyme was calculated. The result is shown in FIG.

Further, in this example, enzyme-substrate reaction was performed using maltose (maltose) instead of sucrose as a substrate solution, and the inhibitory activity of sample E was examined.
That is, a mixed solution obtained by mixing 250 μL of pH 7.0 phosphate buffer, sample E, and 50 μL of enzyme solution was incubated at a temperature of 37 ° C. for 30 minutes. As the enzyme solution, maltase (concentration 1 unit / mL, α-glucosidase manufactured by Wako Pure Chemical Industries, Ltd.), which is a kind of α-glucosidase and has a decomposition activity on maltose, was used. Next, 500 μL of the substrate solution was added to this mixed solution, and incubated at a temperature of 37 ° C. for 20 minutes. As a substrate solution, 37 mM sucrose was used. The reaction was then stopped by immersing in boiling water for 2 minutes and then cooled. Next, 10 μL of this reaction solution was added to 250 μL of glucose reagent, mixed, and incubated at room temperature for 10 minutes. This is designated as test sample X2.
Moreover, the above-mentioned enzyme-substrate reaction was performed without using sample E. Specifically, 250 μL of pH 7.0 phosphate buffer and 50 μL of enzyme solution (1 unit / mL maltase) were mixed and incubated at a temperature of 37 ° C. for 30 minutes. Next, in the same manner as in the case of the test sample X1, the substrate solution (37 mM maltose) was added to this mixed solution and incubated, the reaction was stopped in boiling water, and after cooling, the reaction solution was used as a glucose reagent. In addition, they were mixed and incubated. This is designated as test sample Y2.

Next, also for the test sample X2 and the test sample Y2, the absorbance at a wavelength of 510 nm was measured in the same manner as the test samples X1 and Y1, and the% inhibitory activity (I) was calculated based on the following formula (2). . In this example, the above-described enzyme-substrate reaction was performed while changing the concentration of sample E, and the% inhibitory activity of sample E on maltose-degrading enzyme was calculated. The result is shown in FIG.
I = (Y2−X2) / Y2 × 100 (2)

As a positive control, an enzyme-substrate reaction was performed using acarbose instead of sample E, and the inhibitory activity (% inhibitory activity) of acarbose on maltose-degrading enzyme was measured in the same manner as sample E. In this example, the above-mentioned enzyme-substrate reaction was carried out while changing the concentration of acarbose, and the% inhibitory activity of acarbose on maltose-degrading enzyme was calculated. The result is shown in FIG.
1 and 2 show the results of measuring% inhibitory activity twice for each concentration, and FIGS. 3 and 4 show the results of measuring three times each.

  As is known from FIGS. 1 to 4, it can be seen that the sample E has an inhibitory activity on α-glucosidase (sucrose-degrading enzyme and maltose-degrading enzyme) as in the case of acarbose. Moreover, it turns out that the inhibitory activity with respect to the alpha-glucosidase degrading enzyme of the sample E increases depending on the density | concentration of the sample E.

(Example 2)
This example is an example in which a volunteer subject actually ingests the calorie-restricted food prepared in Example 1 and examines its effect.
Specifically, a different dietary group (sample E), a Salacia extract (sample C1), a Coleus extract (sample C2), and a dextrin (sample C3) for comparison are ingested by seven different groups of subjects. And examined the change in weight and fullness. The sample C1 of Example 1 was used as the Salacia extract, and the sample C2 of Example 1 was used as the Coleus extract. Dextrin was employed as a placebo.

The change in body weight was evaluated by measuring the change in body weight before and after the start of the test with each subject group taking one of the samples E and C1 to C3 for one week. Each sample was infused with 2 capsules each containing 250 mg before breakfast and dinner. The result is shown in FIG.
In addition, the subject may observe the feeling of fullness in the abdomen during the ingestion period, and the result is evaluated in four levels (“very strong”, “strong”, “no change”, “no feeling of fullness”) I was asked to. The results are shown in Table 1.

  As can be seen from FIG. 5, Sample E was confirmed to have an effect of slightly reducing body weight compared to Sample C3 as a placebo. However, since the intake period was a short period of one week, the difference in the effect on the weight change of each sample (sample E, sample C1 to sample C3) was very small.

Further, as is known from Table 1, six people except one out of seven people who took the sample E felt an enhancing effect on the feeling of fullness. On the other hand, about the sample C1 and the sample C2, there were 2 persons who felt the enhancement effect of fullness, and there were 0 persons about the sample C3.
From this, it can be seen that Sample E has an excellent effect of improving fullness as compared with Samples C1 to C3.

An explanatory view showing a bar graph which shows% inhibition activity to sucrose degradation enzyme of sample E concerning Example 1. FIG. BRIEF DESCRIPTION OF THE DRAWINGS Explanatory drawing showing the bar graph which shows the% inhibitory activity with respect to the sucrose degradation enzyme of acarbose concerning Example 1. FIG. Explanatory drawing showing the bar graph which shows the% inhibitory activity with respect to the maltose decomposing enzyme of the sample E concerning Example 1. FIG. BRIEF DESCRIPTION OF THE DRAWINGS Explanatory drawing showing the bar graph which shows the% inhibitory activity with respect to the sucrose degradation enzyme of acarbose concerning Example 1. FIG. Explanatory drawing which shows distribution of the weight change about the test subject concerning Example 2 which ingested each sample (sample E, sample C1-sample C3).

Claims (4)

A calorie-restricted food comprising an inhibitory substance having an inhibitory activity against α-glucosidase as an active ingredient,
A calorie-restricted food comprising a competitive inhibitor and a non-competitive inhibitor as the inhibitor.   2. The calorie-restricted food according to claim 1, wherein the antagonistic substance contains salacinol and / or kotalanol.   3. The calorie-restricted food according to claim 1, wherein the antagonistic inhibitor is a Salacia extract obtained by extracting a root and / or trunk of Salacia reticulata of the Dechinmuraceae or Nishimidae family with a solvent.   4. The calorie according to claim 1, wherein the non-antagonistic inhibitor is a Coleus extract obtained by extracting a root of Coleus forskori, which is a Labiatae plant, with a solvent. Restricted food.

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