JPH09104624A - Alpha-glycosidase inhibitor and composition, sweetener, food and feed composed mainly of sugar and containing the inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject inhibitor having a mild inhibitory action on α-glucosidase, effective for suppressing abrupt increase in blood sugar level to keep the secretion of insulin to a low level and usable for the food for the patient of diabetes. SOLUTION: This inhibiting agent contains a hydroxyproline, i.e., 4-hydroxy-L- proline as an active component. A composition having mild digesting action on sugars and suppressing the insulin secretion to a low level can be produced by combining the α-glucosidase inhibitor with starch or one or more kinds of digestible sugars selected from oligosaccharides originated from starch. The composition is utilizable as a sweetener for diabetic patient, food, health food and feed. The compounding amount of the α-glucosidase inhibitor is 0.5-30wt.% based on the amount of the carbohydrates (sugars) in the food or feed.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention slowly inhibits the action of α-glucosidase, delays the digestion of starch, starch-derived oligosaccharides and sucrose, and as a result suppresses a sharp rise in blood glucose level, The present invention relates to an α-glucosidase inhibitor having an action of suppressing insulin secretion to a low level, a composition containing sugar as a main component, a sweetener, a food and a feed.

[0002]

2. Description of the Related Art Conventionally, when ingesting a diet rich in dietary fiber-rich carbohydrates, absorption of nutrients from the intestine is moderated, post-prandial blood glucose elevation can be suppressed, and insulin secretion can be suppressed to a low level. It has been reported to prevent diabetes [Don't forget fiber by Dr. Denis Burkitt
in your diet (Japanese translation; "Dietary fiber can prevent modern diseases" published by Chuo Koronsha on May 25, 1983. 125
Pp.-137) ", published by Daiichi Shuppan Co., Ltd. on May 15, 1982," Dietary Fiber ", pages 271-286]. However, unlike the sugar that can be freely added to and mixed with a wide range of foods, the above-mentioned dietary fiber is limited in its use for various foods. For example, dietary fiber itself has no sweetness and cannot be used as a sweetener or a sweet material for beverages such as coffee and juice, cakes and sweets.

Further, in recent years, when an α-glucosidase inhibitor is administered, the α-glucosidase inhibitor inhibits the action of α-glucosidase localized in the microvilli of the small intestine, resulting in a rapid rise in postprandial blood glucose level and subsequent insulin level. It is known to suppress a sharp rise (for example, JP-A-52-12).
2342, DIABETIC MEDICINE, 1993; 10: 688-69.
3,134-138, Am, J.Clin.Nutr.1992; 55: 318S-9S, JP-A-57-200335, Am, J.Clin.Nutr.1992; 55:
314S-7S, see JP-A-57-59813). Among such α-glucosidase inhibitors, acarbose is used as an oral diabetes remedy for non-insulin dependent diabetes (abbreviation: NIDDM). However, these substances are essentially foreign substances to living organisms, and there are concerns about their safety, and strict regulations are set for the amount of use.

Further, as indigestible or indigestible oligosaccharides, fructooligosaccharides, galactooligosaccharides, etc., starch hydrolysates and sugar alcohols of other oligosaccharides (for example, sugar alcohols of maltitol and maltooligosaccharides). , Isomaltose and its oligosaccharides, sugar alcohols, reduced palatinose and lactitol) have been conventionally used as sweeteners which hardly cause an increase in blood glucose level due to their indigestibility and low digestibility. However, the indigestible or low-digestible oligosaccharides used as sweeteners with little increase in blood sugar level have the drawback that diarrhea is easily induced if the intake is wrong.

Further, foods and drinks made from Gymnema sylvestre from India, which has an inhibitory effect on the absorption of sugars, for the purpose of suppressing the rise in blood sugar level are disclosed in JP-A-61-5023 and JP-A-63-208532. Has been proposed to.

[0006] The action of these extracts of Gymnema sylvestre and Gymnema minor drum is due to the suppression of the absorption action of sugars, and if the intake amount is incorrect, the blood sugar level will be lowered too much as a side effect, and unabsorbed sugars will become large in the large intestine. Reached
There was a risk of causing diarrhea and other disorders.

JP-A-6-65080 discloses that naturally-occurring saccharides centered on L-arabinose are safe α-
It is said that it can be used as a glucosidase inhibitor.
However, since arabinose and the like are pentoses, browning due to heating is unavoidable, and there is a great limitation in using them by adding them to carbohydrates.

[0008]

As described above, conventional α-glucosidase inhibitors have various problems.
A substance that is contained in the foods that are normally consumed, has excellent processing properties, is absorbed from the digestive tract, and can be effectively used in the body.
-The emergence of glucosidase inhibitors has been desired.

Therefore, the present invention searches for a substance that inhibits α-glucosidase localized in the microvilli of the small intestine, can be used as a food material, a sweetener, and a feed, can prevent diabetes, and An object of the present invention is to provide an α-glucosidase inhibitor suitable for patients, a composition mainly comprising sugar containing the α-glucosidase inhibitor, a food material, a sweetener, and a feed.

[0010]

[Means for Solving the Problems] The present inventors focused on a protein known as one of the three major elements of foods,
It was examined whether the amino acids of the constituents have an α-glucosidase inhibitory action. As a result, it was discovered that only a specific amino acid has a slow inhibitory effect on α-glucosidase, and further, when an amino acid having such an inhibitory effect and an edible carbohydrate or saccharide are used in combination, immediately after ingestion. It was found that it has an effect of suppressing a rapid rise in blood glucose level and suppressing insulin secretion to a low level. Based on such knowledge, the following invention was completed.

That is, the α-glucosidase inhibitor of the present invention is characterized by being an α-glucosidase inhibitor having a slow inhibitory action on α-glucosidase containing hydroxyproline as an active ingredient.

In the present invention, "slow inhibitory effect" means
When the α-glucosidase inhibitor is ingested together with a carbohydrate, the amount of the α-glucosidase inhibitor is 0.5 to 30% by weight relative to the total ingested carbohydrate amount (total sugar mass). , Refers to the effect of appropriately inhibiting the action of the small intestinal digestive enzyme α-glucosidase. A suitable amount of the α-glucosidase inhibitor of the present invention is the conventional α
Since the dose of the -glucosidase inhibitor is large compared to the dose, it can be said that the α-glucosidase inhibitor of the present invention has a "slow inhibitory action".

The term "hydroxyproline" as used herein means
4-hydroxy-L-proline is meant and its stereoisomer, cis-4-hydroxy-L-proline (alias:
Allohydroxyproline) is not meant. Hydroxyproline is a nonessential amino acid enzymatically produced from L-proline in vivo.

Hydroxyproline is one of the important amino acids constituting proteins such as gelatin, casein, globin, etc., and was publicly announced as a food additive on August 10, 1995.

The composition of the present invention mainly composed of a sugar having a slow digestive effect on sugar and an effect of suppressing insulin secretion to a low level is derived from the above-mentioned α-glucosidase inhibitor, sucrose, starch and starch. It is composed of one or more digestive saccharides selected from oligosaccharides. In the composition containing sugar as a main component, the α-glucosidase inhibitor of the present invention is blended so as to be contained in an amount of 0.5 to 30% by weight in the composition containing sugar as a main component. In a composition mainly composed of sugar, the α of the present invention is
-When the glucosidase inhibitor is 0.5% by weight or less,
The inhibitory effect on α-glucosidase is not sufficient, and there is no effect of suppressing a rapid rise in blood glucose level or effect of suppressing insulin secretion to a low level. In addition, in the composition mainly composed of sugar of the present invention, when the content of the α-glucosidase inhibitor of the present invention is 30% by weight or more, the taste peculiar to amino acids may become excessively dominant.

[0016]

BEST MODE FOR CARRYING OUT THE INVENTION The α-glucosidase inhibitor of the present invention is combined with a sugar composed of one or more digestive sugars selected from sucrose, starch and starch-derived oligosaccharides. The sugar-based composition can be used as a sweetener, and such a sweetener has a slow digestive action of sugar when ingested by a human body or an animal, and also has insulin secretion. It has the effect of keeping it low.

When the α-glucosidase inhibitor of the present invention is used by directly adding it to a food composed mainly of carbohydrate, when the food is ingested by a human body or an animal,
The sugar contained in the food is slowly digested, and insulin secretion is suppressed to a low level. Examples of such carbohydrate-based foods include pasta, noodles, breads, cookies, cakes, chocolate, candy, chewing gum, various confectionery, and various soft drinks such as juice and cola. Is mentioned.

When the α-glucosidase inhibitor of the present invention is used for foods having a low or almost no carbohydrate content, for example, coffee, black tea, various teas, etc. Or sweetness mainly composed of saccharides composed of one or more digestive sugars selected from sucrose, starch, and oligosaccharides derived from starch and an α-glucosidase inhibitor in combination It can be used in the form of a charge.

The food to which the α-glucosidase inhibitor of the present invention is added can be health food or food for diabetics. Further, the α-glucosidase inhibitor of the present invention can be added to feed and can be used for diabetes prevention of livestock, pets and the like.

[0020]

【Example】

[Example 1] Example 1 is an example in which hydroxyproline inhibits α-glucosidase in pig small intestine.

20 mM sucrose (S
uc), a 2% soluble starch (S-Starch) solution was prepared. 0.5 ml of each of these was placed in a test tube, and 0.1 M potassium phosphate buffer solution (pH 7.
0-0.4 ml of 100 mM hydroxyproline solution diluted with 0) was added, and 0.4-0 ml of 0.1 M potassium phosphate buffer (pH 7.0) was further added. That is, the total amount of the hydroxyproline solution and the buffer solution was 0.
Buffer solution was added to make 4 ml. To this, add 0.1 ml of the crude enzyme solution of pig small intestine (2-fold dilution) and incubate at 37 ° C for 30
After reacting for 2 minutes, 2M-Tris hydrochloric acid buffer solution (pH
The reaction was stopped by adding 2 ml of 7.0). The amount of glucose generated by the enzymatic reaction was quantified by an enzymatic method using glucose oxidase.

The enzyme solution used in the above enzyme inhibition reaction was the enzyme solution prepared as follows. That is, the mucosa of the inner wall of the small intestine of a pig immediately after being slaughtered was collected, and 5 mM
0.1M potassium phosphate buffer containing EDTA (pH
The mixture was homogenized in 7.0) and the precipitate was collected by centrifugation. This precipitate was suspended in a small amount of the same buffer, and 5 times the same amount of the same buffer containing 1% Triton X-100 was added to the suspension. The mixture was stirred at 0 ° C for 60 minutes to extract the enzyme, and the precipitate was centrifuged to separate the precipitate. After removal of the above, dialysis was carried out in a 0.01 M potassium phosphate buffer (pH 7.0) to obtain a crude enzyme solution.

The results are shown in Table 1 below.

[0024]

[Table 1]

The enzyme activity in Table 1 is represented by the reaction with the substrate alone and the enzyme being 100%. According to Table 1, sucrose (Suc) and soluble starch (S-St)
arch)) as a substrate, as the concentration of hydroxyproline is increased (maximum concentration 40mMo1 / 1),
The result was a decrease in enzyme activity. That is, it can be seen that hydroxyproline inhibits the reaction of the digestive enzyme α-glucosidase (maltase, sucrase, etc.).

Comparative Example 1 In Comparative Example 1, the action of various amino acids on the digestive enzyme α-glucosidase of pig small intestine was examined.

The same substrate (sucrose) and α-glucosidase as those used in Example 1 were used, and it was determined in the same manner as in Example 1 whether various amino acids had an inhibitory effect on the digestive enzymes of pig small intestine. Experimented and investigated. The results are shown in Table 2 (non-essential amino acids) and Table 3 (essential amino acids) below.
Shown in The enzyme activity without addition of various amino acids was defined as 100%, and the specific activity (%) value against this was shown for each addition concentration of various amino acids.

[0028]

[Table 2]

[0029]

[Table 3]

According to Tables 2 and 3, the specific activity is 10 regardless of whether the amino acid is added or not, even if the added amount of the amino acid is increased.
It has been changed at around 0%, and it can be seen that these amino acids have no α-glucosidase inhibitory effect.

Example 2 This Example 2 is an example in which the inhibitory effect on blood glucose level increase and the insulin secretory effect after glucose loading were investigated.

Male Sugague-Dawley rats weighing 150 g were preliminarily bred for 7 to 8 days with a basic diet having the composition shown in Table 4 below. The animals were raised one by one in an apartment-type gauge, and were allowed free access to feed and drinking water. The temperature in the breeding room is 22 ± 1 ° C, and the light period during the breeding period is 7:00 to 19:00.
0 and the dark period were 2 cycles of light and dark every 12 hours from 19:00 to 7:00.

[0033]

[Table 4]

Prior to the implementation, the body weight was measured to exclude individuals having extremely different body weights, and each group was divided into 6 to 7 animals. Before implementation, they were fasted overnight.

The rats of each group were orally administered with the following sugar solution compositions (A, B, and C) (sucrose and / or inhibitor, or physiological saline as a control) using a gastric sonde, and 0 min.
After 5 minutes, 30 minutes, 60 minutes, 120 minutes, and 180 minutes, arterial blood was collected, and the blood glucose concentration (serum glucose amount) and the serum insulin concentration were measured by the enzyme method.

Sugar solution composition: A: 20% sucrose (control) B: 20% sucrose + 1% hydroxyproline C: physiological saline The dosage is 250 m sugar per 100 g of rat body weight.
It was administered at a rate of g, and 2 ml of physiological saline was administered. The time course of blood glucose level after administration is shown in Table 5 below, and the time course of serum insulin amount is shown in Table 6 below.

[0037]

[Table 5]

[0038]

[Table 6]

As is clear from Tables 5 and 6, the blood glucose level and the serum insulin level in the control group reached their maximum values 15 minutes after the administration, and gradually decreased thereafter. This is consistent with the various literature reports reported to date. After 15 minutes in the control group (20% sucrose) in Table 5, the carbohydrate (sucrose) was finally decomposed into glucose in the small intestine by the digestive enzyme α-glucosidase, and glucose was forced into the blood vessel. It is shown that it was taken in and caused a rapid rise in blood glucose level. Further, according to Table 6, the control group at this time showed that insulin secretion was strongly promoted as the blood glucose level rapidly increased. It can be seen that the action of insulin gradually lowers the blood glucose level 15 minutes after administration, and eventually reaches the normal level (Table 5).

On the other hand, in the sucrose solution containing hydroxyproline, the maximum blood glucose level after 15 minutes was clearly lower than that in the control group containing only sucrose, and the blood glucose level after 15 minutes was statistically significant at 5%. There was a significant difference.

Further, as shown in Table 6, the serum insulin concentration in the sucrose solution mixed with hydroxyproline did not increase sharply, and the insulin level after 15 minutes was compared with the control group (20% sucrose). A low value was taken, and a statistically significant difference was observed at a significance level of 1%.

Example 3 In Example 3, starch was used as a sugar and hydroxyproline was mixed as an α-glucosidase inhibitor to suppress the increase in blood glucose level after sugar loading and the effect of suppressing insulin secretion. It is the example which investigated.

The procedure of Example 2 was repeated except that starch was used in place of sucrose. 15 after sugar loading in Example 3
The blood glucose level and insulin concentration after the minutes are shown in Table 7 below.

[0044]

[Table 7]

As is clear from Table 7, when hydroxyproline was mixed with starch and administered orally, the control group (1
It can be seen that both the increase in blood glucose level and the amount of insulin secretion after 15 minutes are inhibited as compared with 5% starch).

[Example 4] When the α-glucosidase inhibitor of the present invention was used in a beverage
Combined Examples Sweeteners were prepared by mixing 1% and 5% of hydroxyproline with sucrose within the quantitative range in which the α-glucosidase inhibitory effect was observed, as shown in the above Examples.

A sample in which only 10% of sucrose was dissolved was used as a control, and a sample in which the same amount of the sweetener was dissolved was used as a test group.

Each coffee solution was trained 10 times.
The name was given to a panelist and a sensory test of its taste was conducted. As a result, 2 out of 10 people felt light bitterness and astringency in the 5% added coffee, and 2 people felt poor sweetness. 1 out of 10 felt a slight bitterness with 1% addition, 1
Three of the 0 people felt that 5% hydroxyproline had a pleasant astringency, and the others answered that there was no difference between the samples. From these results, it can be seen that the α-glucosidase inhibitor of the present invention can be used in beverages.

Example 5 Fruit produced by using the sweetener of the present invention as a sweetening material
Example Using the same sweetener as in Example 5, butter rolls were produced and a sensory test was conducted. Butter rolls were similarly made using sucrose as a control sweetener. The material composition of butter roll using sucrose as a control is 500 g of strong flour, 5 g of salt, 300 ml of milk, 20 g of egg yolk,
Sucrose 40g, butter 40g, raw yeast 25
g, prepared by conventional methods. At this time, the state of primary fermentation was about 5% higher than the control by volume.

The test sweetener was used by adjusting the amount of hydroxyproline to be 6 g, that is, about 10% of the carbohydrate, relative to the control sucrose. The butter roll had the same material composition as above except that it was replaced with a control sucrose.

As a result, 2 out of 10 people answered that they were hard and had a good texture, and no difference was found in taste.

Example 6 Fruit produced by using the sweetener of the present invention as a sweetening material
Example Using the same sweetener as in Example 5, sweet chocolate was produced and a sensory test was conducted.

Sweet chocolate was similarly prepared using sucrose as a control sweetener. 100 g of the bitter chocolate block was pulverized, and a sweetener or sucrose was added to dissolve and solidify. The produced sweet chocolate was subjected to a sensory test as described in Examples 5 and 6 above.
The same method was used, but there was no difference in bitterness in taste.

Cooking example: How to make hard candy To 220 g of sucrose, 87 g of distilled water was added and 177 in 30 minutes
Heated to ° C. Thereafter, the temperature was maintained at 100 ° C., 5 g of hydroxyproline was added in 3 portions, and the mixture was stirred well. Then, it was poured into a mold prepared in advance and allowed to cool. As a result, 249 g of hard candy containing about 2% amino acid could be obtained. The taste and aroma were the same as those of ordinary hard candy.

[0055]

EFFECT OF THE INVENTION Hydroxyproline, which is the active ingredient of the α-glucosidase inhibitor of the present invention, is sucrose,
When used in combination with one or more digestive saccharides selected from starch and starch-derived oligosaccharides, it slowly inhibits the action of α-glucosidase, which is a digestive enzyme of the small intestine, resulting in a rapid blood glucose level. It has the effect of suppressing the elevation of insulin and suppressing the secretion of insulin to a low level.

The α-glucosidase inhibitor of the present invention has a slow inhibitory effect, and a large amount of 0.5 to 30 parts by weight of α-glucosidase is contained in 100 parts by weight of the total amount of carbohydrates (total sugar mass) ingested. Since the inhibitor can be mixed and used, it can be used as a food material or a sweetener that can be added to and mixed with food.

The foods and sweeteners containing the α-glucosidase inhibitor of the present invention can be useful for the prevention of adult diseases including diabetes for healthy people, and for diabetes patients.
It is possible to ease restrictions on sugar intake such as conventional sucrose, starch, and oligosaccharide derived from starch,
It will be possible to provide a wide range of foods and sweeteners suitable for diet.

The feed containing the α-glucosidase inhibitor of the present invention is useful as a feed for preventing diabetes in animals such as pets and livestock.

Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location A61K 47/26 A61K 47/26 L 47/36 47/36 L C12N 9/99 C12N 9/99 (72) Invention Fukumori Yasunori Kita 4-jo Nishi 1-chome, Chuo-ku, Sapporo-shi, Hokkaido Hokuren Agricultural Cooperative Federation (72) Inventor Shiomi ▲ Toku ▼ Huo-ren, Uenoboro 2-chome, Atsubetsu-ku, Sapporo, Hokkaido 1-29 (72) Invention Person Onodera Shuichi Asahi Peace Building 303 7-23, 1-chome, Heiwadori, Shiroishi-ku, Sapporo, Hokkaido

Claims (10)

[Claims] 1. An α containing hydroxyproline as an active ingredient
-Α- having a slow inhibitory effect on glucosidase
Glucosidase inhibitor. 2. From (1) the α-glucosidase inhibitor according to claim 1, and (2) one or more digestive saccharides selected from sucrose, starch and oligosaccharides derived from starch. A composition comprising a sugar as a main constituent, which has a slow digestive effect on the constituent sugar and an effect of suppressing insulin secretion to a low level. 3. The sugar-based composition according to claim 2, wherein the α-glucosidase inhibitor is blended so as to be contained in an amount of 0.5 to 30% by weight in the sugar-based composition. 4. A sweetener comprising a composition mainly comprising sugar according to claim 2 or 3 as an active ingredient, which has a slow digestive effect on sugar and an effect of suppressing insulin secretion to a low level. 5. The sweetener for diabetics according to claim 4. 6. A food having a slow digestive action on sugar and having an action of suppressing insulin secretion to a low level, to which the composition mainly comprising sugar according to claim 2 or 3 is added. 7. A food for diabetic patients, which has a slow digestive action of sugar and has an action of suppressing insulin secretion to a low level, to which the composition mainly comprising sugar of claim 2 or 3 is added. 8. The α-glucosidase inhibitor according to claim 1 is blended in a food containing carbohydrate in an amount of 0.5 to 30% by weight based on the amount of carbohydrate (mass of sugar) in the food. A health food having a slow digestion action of sugar and a low insulin secretion action, which is characterized by the following. 9. The α-glucosidase inhibitor according to claim 1 is blended in a food containing carbohydrate in an amount of 0.5 to 30% by weight based on the amount of carbohydrate (mass of sugar) in the food. A food for diabetics, which has a slow digestion action of sugar and a low insulin secretion action. 10. The α-glucosidase inhibitor according to claim 1 is blended in a feed containing carbohydrates in an amount of 0.5 to 30% by weight based on the amount of carbohydrate (mass of sugar) in the feed. A feed having a slow sugar digestion action and a low insulin secretion action.