JP2015078213A - α-GLYCOSIDASE INHIBITOR - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an α-glycosidase enzyme inhibitor which has high safety, sugar digestion and absorption inhibition, postprandial blood glucose elevation inhibition, and a hyperglycemia improving effect, and to provide a food composition containing the inhibitor, and to provide a pharmaceutical composition.SOLUTION: The invention provides an α-glycosidase enzyme inhibitor comprising an enzyme-treated material of xyloglucan. The xyloglucan is Tamarindus indica seed extract (tamarind seed gum). The enzyme is an α-glycosidase enzyme inhibitor which is a sugar degrading enzyme, such as cellulase, xylase, glucanase, and pectinase.

Description

  The present invention relates to an xyloglucan enzyme-treated product, a method for producing the same, and its use, and more particularly to a glycosidase α-glycosidase inhibitor containing the xyloglucan enzyme-treated product.

  Diabetes mellitus is a disease that is said to affect 180 million people worldwide (according to fiscal 2006), and is on the rise mainly in regions where economic development and population growth are remarkable. And since diabetes may progress to many serious complications, an immediate measure for prevention or treatment of complications is required. Diabetes is caused by insulin-dependent diabetes mellitus (IDDM), which is deficient in blood glucose-regulating hormone due to destruction of β-cells in the pancreas, and by quantitative reduction of insulin or insulin resistance due to symptoms such as obesity. It is classified as type 2 which is non-insulin dependent diabetes mellitus (NIDDM).

  Treatment with insulin administration is performed for type 1 and type 2 end-stage patients. Particularly in the case of type 2, a reduction in the risk of progression of diabetes due to improvement of lifestyle is considered effective. It is effective to appropriately maintain the blood glucose level as an index, but it has been reported that the postprandial blood glucose level is more important as an index than the fasting blood glucose level which has been regarded as important so far.

  Voglibose, Acarbose and the like have been confirmed to have an effect of suppressing blood sugar level increase after meals, and these substances are used as antidiabetic drugs. However, although α-glycosidase inhibitory activity is very strong, there are problems with the strictness and safety of usage. That is, when the above-mentioned therapeutic agent is administered at a high dose, disaccharides (such as surcourse and maltose) whose degradation is inhibited in the small intestine are rapidly degraded by Escherichia coli in the large intestine. Cause various symptoms.

  In order to solve such problems, food-based materials that are easy to adjust for side effects and doses even though their inhibitory action is slow are attracting attention. From such a viewpoint, many studies have been made, such as a composition for suppressing an increase in blood glucose level containing a degradation product of alginic acid (Patent Document 1), an oligosaccharide in which roasted dextrin is acted on with an enzyme (Patent Document 2), panose (patent) Literature 3), maltotriitol (Patent Literature 4) and the like have found α-glycosidase inhibitory activity.

  However, these materials have room for further improvement because they have a low inhibitory activity or cause bitterness or pungent taste when added to food.

Japanese Unexamined Patent Publication No. 2006-193448 JP 2008-297211 JP 2007-084467 A JP 2008-220231 A

  In view of the above situation, the present invention is to provide a material exhibiting α-glycosidase inhibitory activity which is highly safe and can be used for pharmaceuticals, foods and drinks, feeds and the like.

  As a result of diligent research, the present inventors have found that an enzyme-treated product obtained by treating a thickening polysaccharide xyloglucan, preferably tamarind seed gum, used as a food additive with an enzyme with an α-glycosidase inhibitory activity. It was found to have The present invention is based on these findings.

That is, the present invention is as follows.
(1) An α-glycosidase enzyme inhibitor containing an enzyme-treated product of xyloglucan.
(2) The inhibitor according to (1), wherein the xyloglucan is tamarind.
(3) The inhibitor according to (1), wherein the enzyme is a saccharide-degrading enzyme.
(4) A food composition for suppressing digestion and absorption of carbohydrates, suppressing postprandial blood glucose elevation, improving hyperglycemia or anti-obesity, comprising an enzyme-treated product of xyloglucan.
(5) A pharmaceutical composition comprising an enzyme-treated product of xyloglucan.

  According to the present invention, an α-glycosidase enzyme inhibitor comprising an enzyme-treated product of xyloglucan is provided. In addition, according to the present invention, there is provided a food for saccharide digestion and absorption suppression, postprandial blood glucose increase suppression, hyperglycemia improvement or anti-obesity food containing an enzyme-treated product of xyloglucan. The enzyme-treated product of the present invention is useful as a pharmaceutical composition intended for diseases such as diabetes in addition to the above-mentioned food.

Hereinafter, the present invention will be described in detail.
The present invention relates to an enzyme-treated product of xyloglucan.
Xyloglucan is a combination of α-D-xylopyranose and β-D-galactopyranosyl- (1,2) -α-D-xylopyranose linked to glycan residues via α (1,6) linkages. It is characterized by a (1,4) -β-D-glucan main chain having a side chain substituent. The distribution of residues in the side chain is different in different types of xyloglucans. Three oligomeric units have been identified in the xyloglucan structure, namely monosaccharides, octasaccharides and heptasaccharides (with different numbers of galactose side chains from each other) (U. Hiroshi, Trends in Glycoscience and Glycotechnology, 14, 355-). 376, 2002).

Examples of the xyloglucan used in the present invention include tamarind seed extract (tamarind seed gum), Detarium senegalense Gmelin extract, and Afzelia africana extract, and tamarind seed gum is more preferable.
The extract of tamarind seed gum can be obtained by subjecting the seed or the endosperm portion of the seed to extraction with warm to hot water or an alkaline aqueous solution.
Moreover, the extract of Detarium senegalense Gmelin can be obtained by extracting the seed or the endosperm portion of the seed with warm to hot water or an alkaline aqueous solution.
The extract of Afzelia africana can be obtained by subjecting the seed or the endosperm portion of the seed to extraction with warm to hot water or an alkaline aqueous solution.

In the present invention, the xyloglucan enzyme-treated product is obtained by enzymatic treatment of xyloglucan. Therefore, the xyloglucan enzyme-treated product of the present invention includes a xyloglucan enzyme-treated product, a xyloglucan crushed product, or a powder thereof.
“Enzyme treatment” is to decompose, change the composition, or change the structure of the components by allowing xyloglucans and plants, algae, pulverized microorganisms or the like containing xyloglucans to act on the enzyme.

The enzyme used in the present invention is selected from those that act on biopolymers. Biopolymers include polysaccharides, proteins, nucleic acids and the like. As enzymes that act on biopolymers, saccharolytic enzymes, proteolytic enzymes, and nucleolytic enzymes can be used. Glycosidases such as polysaccharide degrading enzymes are preferred in that they act directly on the sugar structure. Examples of polysaccharide-degrading enzymes include cellulase, xylase, glucanase, pectinase and the like, but are not limited thereto. In addition, even enzymes that are considered not to act directly on the sugar structure are useful because they can produce sugar components that express their functions by degrading other biopolymers. Examples of such enzymes include proteases and nucleases.
In the treatment for obtaining the enzyme-treated product of xyloglucan of the present invention, in addition to the enzyme, salts that adjust ionic strength, buffers or water that adjust pH are used as necessary. The enzyme treatment can be obtained by reacting xyloglucan with a polysaccharide-degrading enzyme at 10 to 70 ° C. for 1 to 100 hours, but is not limited to these methods, and those skilled in the art can use known methods. It can be selected appropriately. The xyloglucan enzyme-treated product of the present invention may be in any form such as an enzyme reaction solution, a concentrated solution thereof, a solid obtained by drying the concentrated solution, or a powder thereof.

For example, 2.5 kg of tamarind is crushed, 100 ml of polysaccharide degrading enzyme is added to the crushed material, and the mixture is reacted at 40 ° C. for 72 hours. This operation is repeated as necessary, and the resulting reaction solution is combined and concentrated and dried to prepare an enzyme-treated product.
Furthermore, purified xyloglucan can also be used. For example, xyloglucan can be purified by crushing tamarind, extracting with hot water, and then precipitating polysaccharide components with alcohols such as ethanol and isopropanol. Purified xyloglucan can be similarly reacted with an enzyme.

  In the present invention, the xyloglucan-treated product can be further isolated and purified by repeating the fractionation operation by various types of chromatography such as ion exchange chromatography, reverse phase chromatography, and gel filtration chromatography. However, isolation and purification are not limited to these methods. In addition, α-glycosidase activity can be used as an index during the fractionation, isolation and purification steps. Furthermore, if desired, the action of inhibiting digestion and absorption of saccharides, the action of inhibiting the increase in blood sugar level, and the like can also be employed as indicators for confirming the properties of the xyloglucan enzyme treated product of the present invention.

The present invention relates to an α-glycosidase enzyme inhibitor containing an enzyme-treated product of xyloglucan as an active ingredient.
According to the present invention, a new mechanism of action of an xyloglucan enzyme-treated product is found, and the present invention provides a novel use of the xyloglucan enzyme-treated product.

The processed xyloglucan enzyme means a substance (antagonist) that inhibits the enzyme activity of α-glycosidase, that is, the action of cleaving a glycoside bond of a specific sugar. It can be said that the antagonist of α-glycosidase enzyme has an action of suppressing the action of α-glycosidase to cleave the glycosidic bond of sugar by acting on α-glycosidase.
Here, α-glycosidase refers to a sucrase (also referred to as invertase) that decomposes sucrose into glucose and fructose and a maltase that decomposes maltose into glucose.
The inhibitory activity of α-glycosidase (schuclase inhibitory activity and maltase inhibitory activity) can be measured, for example, as follows.

  A commercially available acetone powder of rat small intestinal mucosal enzyme is homogenized in a buffer solution, and the centrifuged supernatant is used as the enzyme solution. A sample that was reacted with a substrate (sucrose or maltose) and a sample, respectively, at 37 ° C, measured the production glucose to determine its production rate, and performed the same reaction as above without adding an enzyme treatment solution The inhibition rate is calculated assuming that the amount of glucose produced is 100.

  In the present invention, an enzyme inhibitor of α-glycosidase can also be used as a research reagent related to sugar metabolism such as an effect of increasing blood glucose level after meals. Examples of the method for evaluating the α-glycosidase enzyme inhibitory activity include, but are not limited to, evaluation of sugar metabolism in intestinal epithelial cells.

The amount of xyloglucan enzyme-treated product contained in the α-glycosidase enzyme inhibitor of the present invention can be appropriately set by those skilled in the art. For example, when used for measurement of maltase inhibitory activity, a unit usage standard 1 μg to 10 g, preferably 5 μg to 1 g, more preferably 10 μg to 100 mg.
Moreover, the enzyme-treated product of xyloglucan of the present invention can be used as various functional foods.
For example, the enzyme-treated product of the present invention is used as a composition for inhibiting digestion and absorption of carbohydrates, suppressing postprandial blood sugar elevation, improving hyperglycemia, or anti-obesity food.

  Inhibition of digestion and absorption of carbohydrates means that carbohydrates ingested from the diet or that have been affected by digestive enzymes pass through the digestive tract such as the small intestine without being absorbed, and in particular, penetrate the small intestine. It is said that digestion and absorption were suppressed when the sugar mass was reduced.

Postprandial blood sugar elevation is the ingestion of sugars such as starch, then it is broken down into disaccharides in the oral cavity and intestinal tract, and is broken down by disaccharide-degrading enzymes present in the intestinal epithelium to permeate the intestinal tract It means that the blood glucose level rises by shifting to, and when the blood glucose level suddenly increases around 2 hours after the meal, the postprandial blood glucose rises.
Inhibition of postprandial blood glucose increase means suppression of a rapid increase in postprandial blood glucose level, and means that the blood glucose level gradually increases around 2 hours after the meal.

  The composition containing the enzyme-treated product of xyloglucan of the present invention can be a health food, a functional food, a nutritional supplement, a beverage, or an additive thereof, in addition to a general processed food. As an aspect of the food of the present invention, the dried powder, extract or purified product of the enzyme-treated product of xyloglucan of the present invention is mixed with food as it is, or as a liquid, gel, powder or solid food Can be used. Examples of these food products include beverages, dairy products, seasonings, noodles, processed meat and fish processed foods, margarine, bread, and confectionery. The food of the present invention covers a wide variety of forms and is not limited to the above examples, but from the viewpoints of suppressing blood sugar elevation, improving hyperglycemia, and anti-obesity, the form of dietary supplement and health food is preferred.

  In the α-glycosidase enzyme inhibitor and the pharmaceutical composition of the present invention, the xyloglucan enzyme-treated product may be used as it is, or it may be formulated or made into food by a pharmaceutically acceptable carrier. Moreover, the form which added the xyloglucan enzyme processed material to the alcoholic beverage, the soft drink, etc. may be sufficient as the xyloglucan enzyme processed material of this invention. For formulating, specifically, various organic or inorganic carrier substances commonly used as a formulation material are used. The α-glycosidase enzyme inhibitor of the present invention and the xyloglucan enzyme-treated product contained in the pharmaceutical composition can be used alone, but may form a salt, and the salt of the candidate substance is physiologically acceptable. Salts with acids (for example, inorganic acids and organic acids) or bases (for example, alkali metals) are used, and physiologically acceptable acid addition salts are preferred. Examples of such salts include salts with inorganic acids (eg, hydrochloric acid, phosphoric acid, hydrobromic acid, sulfuric acid, etc.), or organic acids (eg, acetic acid, formic acid, propionic acid, fumaric acid, maleic acid, And salts with succinic acid, tartaric acid, citric acid, malic acid, succinic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, and the like.

  The α-glycosidase enzyme inhibitor and pharmaceutical composition of the present invention can be formulated according to a conventional method, and the formulation may be solid or liquid. For example, capsules, granules, emulsions, tablets, pills, Dragees, solutions, gels, syrups, slurries, suspensions, and the like, may include pharmaceutically acceptable carriers.

  In solid preparations, excipients, lubricants, binders, disintegrants, thickeners and the like are used. In liquid preparations, solvents, dispersants, solubilizers, suspending agents, isotonic agents, Buffering agents, soothing agents, etc. are used. In addition, additives such as preservatives, antioxidants, colorants, sweeteners and the like can be used as necessary.

  The pharmaceutical composition of the present invention is, for example, an oral administration form such as tablets, capsules, granules, powders, drinks, etc., and intravenous drip or injection, or intramuscular injection, enteral administration, etc. It can be administered in the form. Administration may be either systemic or local.

  EXAMPLES Hereinafter, although the Example demonstrates the implementation method of this invention further in detail, these Examples aim at the illustration of this invention, and do not limit this invention.

(1) Preparation of xyloglucan enzyme-treated product Suspended in an autoclave at 121 ° C. for 20 minutes after suspending 2.5 g of tamarind seed gum (manufactured by MRC polysaccharides) in 100 mM sodium phosphate buffer (pH 7.0). did. Dispense 9 ml of tamarind seed gum solution into a test tube and cool to room temperature. Prepare 1 ml of the enzyme solution prepared in advance in the concentration shown in Table 1 and sterilized with a 0.2 μm filter. The mixture was added and reacted at 40 ° C. for 72 hours.

  After the reaction, autoclaving was performed at 121 ° C. for 20 minutes in order to inactivate the enzyme, and then α-glycosidase inhibitory activity was measured.

(2) α-Glycosidase (maltase) inhibitory activity measurement Preparation of small intestine powder enzyme 0.1 M sodium phosphate buffer (pH 7) was added to rat small intestine acetone powder (SIGMA), and 100 mg / ml. The supernatant obtained by centrifugation (5,000 × g, 15 minutes) was used as a crude enzyme solution.

α-Glycosidase inhibitory activity measurement (maltose substrate)
Add 100 μl of 0.1 M sodium phosphate buffer (pH 7.0), 50 μl of 2% maltose solution, and 800 μl of tamarind enzyme treatment solution prepared in (1), and add 50 μl of the crude enzyme solution prepared in (1). And reacted at 37 ° C. for 120 minutes.
(Sucrose substrate)
Add 100 μl of 0.1 M sodium phosphate buffer (pH 7.0), 50 μl of 4% sucrose solution, and 800 μl of the tamarind enzyme treatment solution prepared in (1), and add 50 μl of the crude enzyme solution prepared in (1). And reacted at 37 ° C. for 120 minutes.
(Isomaltose substrate)
Add 100 μl of 0.1 M sodium phosphate buffer (pH 7.0), 50 μl of 5% isomaltose solution, and 800 μl of the tamarind enzyme treatment solution prepared in (1), and add 50 μl of the crude enzyme solution prepared in (1). And it was made to react at 37 degreeC for 120 minutes.
(Soluble starch substrate)
Add 100 μl of 0.1 M sodium phosphate buffer (pH 7.0), 50 μl of 1% soluble starch solution, and add 800 μl of tamarind enzyme treatment solution prepared in (1), add 50 μl of crude enzyme solution prepared in (1). And it was made to react at 37 degreeC for 120 minutes.

The glucose concentration of the reaction solution was measured using a glucose measurement kit (D-Glucose (GOPOD Format) (manufactured by Megazyme)), and the same amount of operation was performed without adding the tamarind enzyme treatment solution. The inhibition rate was calculated as (Table 2).
As a comparative example, L-arabinose or D-xylose added at a final concentration of 0.1 (w / v)% instead of the above-described tamarind enzyme treatment solution was also performed at the same time.

(1) Preparation of xyloglucan enzyme-treated product was performed in the same manner as in Example 1 using tamarind seed gum.
(2) The maltase inhibitory activity was measured according to the case where maltose described in Example 1 was used as a substrate (Table 3).

  From Table 3, maltase inhibitory activity was found in the enzyme-treated product of tamarind-derived xyloglucan, and it was shown to exhibit higher inhibitory activity than L-arabinose and D-xylose used as controls.

According to the present invention, an enzyme-treated product of xyloglucan is provided. The enzyme-treated product of the present invention can be used as an α-glycosidase inhibitor or as various functional foods or pharmaceutical compositions.

Claims (5)

An α-glycosidase enzyme inhibitor containing an enzyme-treated product of xyloglucan. The inhibitor according to claim 1, wherein the xyloglucan is tamarind. The inhibitor according to claim 1, wherein the enzyme is a saccharide-degrading enzyme. A food composition for inhibiting digestion and absorption of carbohydrates, suppressing postprandial blood glucose elevation, improving hyperglycemia, or anti-obesity, comprising an enzyme-treated product of xyloglucan. A pharmaceutical composition comprising an enzyme-treated product of xyloglucan.