WO2006085523A1

WO2006085523A1 - Composition inhibiting rise in blood glucose level

Info

Publication number: WO2006085523A1
Application number: PCT/JP2006/302047
Authority: WO
Inventors: Naofumi Kitabatake; Tetsuya Masuda; Hiroko Naito
Original assignee: Kyoto University
Priority date: 2005-02-09
Filing date: 2006-02-07
Publication date: 2006-08-17
Also published as: JPWO2006085523A1

Abstract

A composition for inhibiting a rise in the blood glucose level characterized by containing, as the active ingredient, a substance selected from among basic proteins such as thaumatin, cytochrome C, lysozyme, ribonuclease and monelin and decomposition products thereof, in particular, thaumatin.

Description

Specification

Composition for inhibiting increase in blood glucose level

Technical field

[0001] The present invention relates to a composition for suppressing an increase in blood glucose level, and more specifically, a pharmaceutical comprising a basic protein such as thaumatin as an active ingredient, particularly suppressing an increase in blood glucose level after ingestion of carbohydrates. , Food-like compositions.

Background art

[0002] Conventionally, various peptides, proteins, polyphenols, dietary fibers, and the like are known as materials that suppress the increase in blood glucose level. For example, indigestible dextrin, guava leaf polyphenol, wheat albumin, L- Specified health foods that contain arabinose, bean drum extract, etc. as active ingredients for the purpose of suppressing an increase in blood glucose levels have already been put on the market.

On the other hand, thaumatin is known as a sweet protein that is contained in the fruits of fruit trees planted in Africa, and has recently been used as a food additive for low-calorie, high-sweetness sweeteners. Have been developed (see, for example, Patent Documents 1, 2 and 3).

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However, there is no report suggesting that thaumatin has an effect of suppressing an increase in blood glucose level after intake of carbohydrates. In addition, there are no reports suggesting an inhibitory action on the increase in blood glucose level of basic proteins including thaumatin.

Patent Document 1: Japanese Unexamined Patent Publication No. 2000-197462

Patent Document 2: Japanese Patent Laid-Open No. 2000-270809

Patent Document 3: Japanese Unexamined Patent Application Publication No. 2004-129595

Disclosure of the invention

Problems to be solved by the invention

[0003] The main object of the present invention is to provide a composition for suppressing an increase in blood glucose level as a novel use of thaumatin and other basic proteins.

Means for solving the problem

[0004] The present inventors are not aware of the mechanism of action and receptors of thaumatin as a sweet protein. Through extensive research, we have found that oral administration of thaumatin strongly suppresses the increase in blood glucose level after ingestion of dalcose in mice and humans. In addition, the inventors have

Other basic proteins like cytochrome c lysozyme have similar effects

And found that its action is persistent.

[0005] The present invention has been completed based on these findings,

(1) A composition for suppressing an increase in blood glucose, comprising as an active ingredient a basic protein having an isoelectric point of about 9 or more or a degradation product thereof having an inhibitory action on an increase in blood glucose,

(2) Basic protein strength The composition according to the above (1), wherein thaumatin, cytochrome C, lysozyme, ribonuclease and monelinka are also selected,

(3) The composition according to the above (1), wherein the basic protein is thaumatin,

(4) The composition according to (1) above, which is a pharmaceutical product,

(5) The composition according to the above (1), which is a food,

(6) Use of a basic protein selected from thaumatin, cytochrome C, lysozyme, ribonuclease and monelin for producing a composition for suppressing an increase in blood glucose level,

(7) The use according to (6) above, wherein the basic protein is thaumatin, and

(8) It provides thaumatin, cytochrome C, lysozyme, ribonuclease or monelin for suppressing an increase in blood glucose level.

The invention's effect

[0006] Saumatine used as an active ingredient of the yarn composition for suppressing an increase in blood glucose level of the present invention is an extremely low-toxic food additive, cytochrome C is a biological component, and lysozyme has little side effects and is orally administered. Known as an enzyme that can be ingested, V is a highly safe substance, and the composition for suppressing an increase in blood glucose level of the present invention contains a relatively small amount before, simultaneously with, or after the intake of saccharides. Oral administration can safely and significantly suppress an increase in blood glucose level.

For example, the amount of a known blood sugar level increase inhibiting ingredient per ingestion is: indigestible dextrin number g, guava leaf polyphenol 0.2 g, wheat albumin 125 mg L-arabinose 180 mg, soybean drum extract 0.3 g In contrast, thaumatin sharply suppresses an increase in blood glucose levels at several mg. Brief Description of Drawings

FIG. 1 is a graph showing the effect of thaumatin on the change in blood glucose level over time in the mouse of Test Example 1.

FIG. 2 is a graph showing the effect of thaumatin on the change in blood glucose level over time in humans of Test Example 2.

FIG. 3 is a graph showing the effect of thaumatin administration time on the time-dependent change in blood glucose level in the mouse of Test Example 3.

FIG. 4 is a graph showing the effect of cytochrome C on the time course of blood glucose level in the mouse of Test Example 4.

FIG. 5 is a graph showing the effect of lysozyme on the time course of blood glucose level in the mouse of Test Example 5.

FIG. 6 is a graph showing the effect of thaumatin on the time course of blood glucose level in the human of Test Example 6.

FIG. 7 is a graph showing the effects of lysozyme, monelin, ribonuclease, thaumatin, and cytochrome C on the time course of blood glucose level in the mouse of Test Example 7.

FIG. 8 is a graph showing the dose-dependent effect of thaumatin on the change in blood glucose level over time in the mouse of Test Example 8.

FIG. 9 is a graph showing the glucose absorption inhibitory effect of thaumatin in Test Example 9 in human small intestine model cells.

FIG. 10 is a graph showing the persistence of cytochrome C blood glucose level increase inhibitory effect in the mouse of Test Example 10.

BEST MODE FOR CARRYING OUT THE INVENTION

[0008] Examples of the basic protein having an isoelectric point of about 9 or more used as an active ingredient of the composition for suppressing an increase in blood glucose level of the present invention include thaumatin (isoelectric point 11.7), cytochrome C (isoelectric point). Point 10), lysozyme (isoelectric point 11), ribonuclease (isoelectric point 9.5), monelin (isoelectric point 11), etc., and thaumatin is preferred. These can be used as commercially available products.

In addition, as a degradation product of these basic proteins having an action to suppress an increase in blood sugar level, For example, hydrolyzed products under mild conditions (for example, pH 7.0, 0.02% trypsin added and reaction at 37 ° C for 30 minutes) can be mentioned.

The compounding amount of the active ingredient in the composition for suppressing an increase in blood glucose level of the present invention is not particularly limited and can be appropriately selected depending on the dosage form of the composition, etc., but is generally 0.005 to 95 with respect to the total amount of the composition. It is about 0.1% by weight, preferably about 0.01 to 80% by weight, and is not particularly limited. Usually, for adults (with a body weight of 60kg), 0.1 to 1 time: L00mg, preferably 1 to: An amount capable of administering or ingesting an active ingredient of about LOmg.

[0009] The composition for suppressing an increase in blood glucose level of the present invention is a liquid, semi-solid or solid pharmaceutical product and food and drink suitable for oral administration or ingestion directly or by dilution or division, etc., according to a conventional method. It can be made the form. Examples of the liquid dosage form include a solution, a syrup, an emulsion, a suspension, and the like. Examples of the semi-solid dosage form include a paste. Examples of the solid dosage form include, for example, Examples include pharmaceutical forms such as powders, granules, tablets, and capsules, and various food forms.

If desired, the composition for suppressing an increase in blood glucose level of the present invention is within a range not violating its purpose. For example, a carrier, an excipient, a diluent, a binder, a disintegrant, a surfactant, a lubricant, a colorant. Further, it may contain known additives such as antioxidants, preservatives, seasonings, and fragrances.

[0010] The composition for suppressing an increase in blood glucose level of the present invention obtained by force can be used as a pharmaceutical or a food such as a food for specified health use, and for example, as a pharmaceutical together with other pharmaceutically active ingredients. In addition, various foods and drinks that are used as a food additive for suppressing blood sugar level elevation together with other edible materials can be used. Such pharmaceuticals and foods and drinks are also within the scope of the present invention.

The composition for suppressing an increase in blood glucose level according to the present invention is, for example, as described above in the same manner as usual pharmaceuticals and foods and drinks, including the case of about 30 minutes before the meal and about 30 minutes after the meal. Oral administration or ingestion to humans who need to suppress the increase in blood glucose level in the dosage range can safely and sharply suppress the increase in blood glucose level due to meals. The value increase effect is persistent. If necessary, the dose can be adjusted in terms of body weight and used to suppress the increase in blood glucose level of other mammals (eg, monkeys, dogs, cats, etc.).

The ability to further explain the present invention with reference to test examples and examples is as follows. It is not limited to that.

Test example 1

[0011] Two groups of 10-week-old male ICR mice (weighing 30-35 g, two per group) were each purified water 200 μ1 [dissolved gnole = 3-ose 100 mg, and purified water 200 μ 1 [dissolved gnoleose 1 OO mg and thaumatin 240 g were orally administered, and blood glucose level was measured over time. The results are shown in Figure 1. In FIG. 1, the vertical axis represents blood glucose level (mgZdl), and the horizontal axis represents elapsed time (minutes) after administration. When thaumatin is administered together with glucose as shown in Fig. 1, the increase in blood glucose level is sharply suppressed.

Test example 2

[0012] Three humans (adults, body weight 50-65 kg) were each orally ingested with 30 g of glucose dissolved in 250 ml of purified water, 5.74 mg of thaumatin only, and 30 g of glucose and 5.74 mg of glucose. Was measured over time. The measurement of blood glucose level was started 15 minutes before ingestion, and the amount of change in blood glucose level at each elapsed time from the blood glucose level 15 minutes before ingestion is shown in FIG. 2 (average of 2 intakes each). In Fig. 2, the vertical axis represents the amount of change (mgZdl) and the horizontal axis represents the elapsed time (minutes).

As shown in Fig. 2, even in humans, the ingestion of thaumatin together with glucose sharply suppresses the increase in blood glucose level.

Test example 3

[0013] Three groups of 8-week-old male C57BL6 mice (body weight 20-25g, 2 per group), glucose dissolved in purified water 1201, 3.3mgZg body weight (glucose administration group), purified Saumatine 10 gZg body weight dissolved in water 1201 and glucose dissolved in purified water 1201 3.3 mgZg body weight and thaumatin 10 μg Zg body weight (somatin simultaneous administration group) were orally administered, and blood glucose level was measured over time. In the thaumatin-only administration group, glucose 3.3 mg / g body weight dissolved in purified water 120 1 was further administered 15 minutes later (the thaumatin pre-administration group). The results are shown in Figure 3. In FIG. 3, the vertical axis represents blood glucose level (mgZdl), and the horizontal axis represents elapsed time (minutes) after administration.

As is apparent from FIG. 3, when thaumatin is administered before glucose administration, the increase in blood glucose level is remarkably suppressed, and thaumatin can also prevent an increase in blood glucose level. Test example 4

[0014] Two groups of 8-week-old male C57BL6 mice (body weight 20-25 g, 2 per group), glucose dissolved in purified water 120 1, 3.3 mg / g body weight, and purified water 120, respectively Glucose dissolved in 1, 3.3 mgZg body weight and cytochrome C5 gZg body weight were orally administered, and blood glucose level was measured over time. The results are shown in Fig. 4. In Fig. 4, the vertical axis represents blood glucose level (mgZdl), and the horizontal axis represents elapsed time (min) after administration.

When cytochrome C is administered together with glucose as shown in Fig. 4, the increase in blood glucose level is sharply suppressed.

Test Example 5

[0015] Two groups of 8-week-old male C57BL6 mice (body weight 20-25g, 8 per group), glucose dissolved in purified water 1201, 3.3mg / g body weight, and purified water 120, respectively Glucose dissolved in 1, 3.3 mgZg body weight and lysozyme 5 gZg body weight were orally administered, and blood glucose level was measured over time. The results are shown in FIG. In FIG. 5, the vertical axis represents blood glucose level (mgZdl), and the horizontal axis represents elapsed time (minutes) after administration.

When lysozyme is administered together with glucose as shown in Fig. 5, an increase in blood glucose level is suppressed.

Test Example 6

[0016] Five humans (adult, body weight 50-65 kg) were each orally ingested with 30 g of glucose dissolved in 250 ml of purified water, and 30 g of glucose and 15 mg of thaumatin, and blood glucose levels were measured over time. Blood glucose measurement starts 15 minutes before ingestion, and blood glucose levels at each elapsed time are shown in Fig. 6. In FIG. 6, the vertical axis represents blood glucose level (mgZdl) and the horizontal axis represents elapsed time (minutes). As shown in Fig. 6, even in humans, the ingestion of thaumatin together with glucose sharply suppresses the increase in blood glucose level.

Test Example 7

[0017] 6 groups of 7-8 week old male C57BL6 mice (body weight 20-25g, 1 group 7 tails), glucose dissolved in purified water 120μ1, 3.3mgZg body weight (glucose administration group), Purified water 1 Glucose dissolved in 20 μ 1 3.3 mg / g body weight and lysozyme 9 μ g / g body weight (lysozyme administration group), glucose dissolved in purified water 120 1 3.3 mg Zg body weight and monelin 9 gZg Body weight (monelin administration group), glucose dissolved in 120 μ1 of purified water 3.3 mgZg body weight and ribonuclease 9 μg Zg body weight (ribonuclease administration group), glucose dissolved in 120 μ1 of purified water 3.3 mgZg body weight and thaumatin 9 GZg body weight (thaumatin administration group), glucose dissolved in 120 μl of purified water 3.3 mgZg body weight and cytochrome C 9 μg Zg body weight (cytochrome C administration group) were orally administered, and blood glucose levels were measured over time. Based on the blood glucose level at the time of administration, the area under the blood concentration curve up to 90 minutes after administration was also calculated. The results are shown in Fig. 7. In FIG. 7, the vertical axis represents the ratio (%) to the area under the blood concentration curve in the glucose administration group.

When lysozyme, monelin, ribonuclease, thaumatin, and cytochrome C are administered together with glucose as shown in Fig. 7, each increase in blood glucose level is sharply suppressed.

Test Example 8

[0018] Five groups of male C57BL6 mice aged 7 to 8 weeks (20 to 25 g body weight, 9 to 10 fish per group) were each dissolved in 120 μ 1 of purified water 3.3 mg Zg body weight (glucose administration group) ), Glucose dissolved in purified water 120 μ1 3.3 mg Zg body weight and thaumatin 0.9 μg Zg body weight (sodium maize 0.9 g administration group), glucose dissolved in purified water 120 1 3.3 mg Zg body weight and thaumatin 1. 8 gZg body weight (somatin 1.8 g administration group), glucose dissolved in purified water 120 1 3.3 mgZg body weight and thaumatin 9. Ο / z gZg body weight (thomatin 9. administration group), purified water 120 1 Glucose 3.3 mgZg body weight and thaumatin 18 gZg body weight (Thomatin 18 g administration group) were orally administered, and blood glucose level was measured over time. The results are shown in Fig. 8. In FIG. 8, the vertical axis represents blood glucose level (mgZdl) and the horizontal axis represents elapsed time (minutes).

As is apparent from FIG. 8, when thaumatin is administered together with glucose, the effect of suppressing the increase in blood glucose level is increased in a dose-dependent manner.

Test Example 9

[0019] In order to investigate the effect of thaumatin on the glucose transport mechanism at the cellular level, the uptake of dalcose when thaumatin was added was examined using a Caco-2 cell monolayer culture system, a human small intestine model. . Caco-2 cells, a human colon adenocarcinoma-derived intestinal epithelial cell line, were cultured on a permeable membrane to form a cell monolayer and used for experiments. For glucose uptake experiments, Caco-2 cell monolayers were prepared using glucose-free HEPES buffered salt soluti. Incubate for 15 minutes on (HBSS), then replace with HBSS (5 mM glucose added) to which thaumatin is added (0 .: LM and 100 μΜ) and ³ H-labeled Darcos solution. 3 ml (: CiZml) was taken up for 30 minutes. Thereafter, glucose permeated through the Caco-2 cell monolayer was measured with a liquid scintillation counter. The result is shown in FIG. In FIG. 9, the vertical axis represents the percentage (%) with respect to the amount of glucose uptake when thaumatin is not added.

As shown in FIG. 9, thaumatin suppresses glucose uptake in the small intestine. Test Example 10

[0020] Two groups of 7-8 week old male C57BL6 mice (body weight 20-25g, 2 per group), glucose dissolved in 120 μl of purified water, 3.3mgZg body weight (glucose administration group), Glucose dissolved in 20 μ 1 of purified water 3.3 mgZg body weight and cytochrome C 9 μg Zg body weight (cytokine C administration group) were orally administered, and blood glucose levels were measured over time. In addition, 2 groups of 7-8 week old male C57BL6 mice (body weight 20-25 g, 1 group 1-2 fish), glucose dissolved in 120 μl of purified water 3.3 mgZg body weight and cytochrome CS / z gZg Body weight was orally administered, and 1 day after (cytochrome C administration, 1 day after glucose administration group) and 6 days after (cytochrome C administration, 1 day after glucose administration group) g Body weight was orally administered, and blood glucose level was measured over time. The result is shown in FIG. In Fig. 10, the vertical axis represents blood glucose level (mg / dl) and the horizontal axis represents elapsed time (minutes).

As is clear from FIG. 10, the effect of cytochrome C to suppress the increase in blood glucose level persists for several days after administration.

Example 1

[0021] Thomatin 5. 74 mg and an appropriate amount of flavor are dissolved in 250 ml of purified water, and sterilized and filtered to produce a liquid composition for suppressing blood glucose elevation directly for drinking.

Example 2

[0022] Cytochrome ClOmg and an appropriate amount of flavor are dissolved in 150 ml of purified water, and sterilized to produce a liquid composition for suppressing blood sugar level elevation that is directly used for drinking.

Example 3

[0023] Lysozyme 10mg and appropriate amount of perfume are dissolved in 150ml of purified water, and sterile filtered directly. A liquid composition for suppressing blood sugar level elevation for use in drinking is produced.

Example 4

[0024] Ribonuclease lOmg and an appropriate amount of fragrance are dissolved in 150 ml of purified water, and sterilized by filtration to produce a liquid composition for suppressing blood sugar level elevation that is directly used for drinking.

Example 5

[0025] Monellin lOmg and an appropriate amount of perfume are dissolved in 150 ml of purified water, and sterilized to produce a liquid composition for suppressing an increase in blood glucose level that is directly used for drinking.

Industrial applicability

[0026] According to the present invention, it is possible to provide a pharmaceutical product and a food product for suppressing blood sugar level elevation that are safe and exhibit excellent blood sugar level elevation inhibiting effect after carbohydrate intake.

Claims

The scope of the claims

[1] A composition for suppressing an increase in blood glucose level, comprising as an active ingredient a basic protein having an isoelectric point of about 9 or higher or a degradation product thereof having an inhibitory action on an increase in blood glucose level.

[2] The composition according to claim 1, wherein the basic protein is selected from thaumatin, cytochrome C, lysozyme, ribonuclease and monelinka.

[3] The composition according to claim 1, wherein the basic protein is thaumatin.

[4] The composition according to claim 1, which is a pharmaceutical product.

[5] The composition according to claim 1, which is a food.

[6] Use of a basic protein, which is also selected from thaumatin, cytochrome C, lysozyme, ribonuclease and monelin, for producing a composition for suppressing an increase in blood glucose level.

7. The use according to claim 6, wherein the basic protein is thaumatin.

[8] Saumatine, cytochrome C, lysozyme, ribonuclease or monelin for suppressing blood sugar level elevation.