JP2003146900A - Sugar breakdown enzyme inhibitor and food containing the inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a sugar breakdown enzyme inhibitor having excellent saccharide breakdown enzyme inhibiting activity and, even if ingested as food, safe for human body and causing no adverse effect, and to provide a food containing the sugar breakdown enzyme inhibitor, effectively suppressing hyperglycoplasmia after eating and, thereby, preventing and improving diseases such as diabetes, corpulence, hyperlipemia and the like. SOLUTION: This sugar breakdown enzyme inhibitor is characterized by containing mate leaf extract obtained by extracting mate leaves as an active component and the food containing the sugar breakdown enzyme inhibitor is provided.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a glycolytic enzyme inhibitor and foods containing the same, and more specifically, a glycolytic enzyme containing a mate leaf extract obtained by extracting mate leaves as an active ingredient. The present invention relates to an inhibitor and a food containing the inhibitor and useful for preventing, treating and improving hyperglycemia such as diabetes, obesity and hyperlipidemia.

[0002]

2. Description of the Related Art In Japan, the eating habits have become rich, and it is now called the age of satiety. Due to excessive caloric intake and lack of exercise, obesity or diabetes is increasing. Currently, one in eight adult males and one in six adult females are obese. In addition, diabetic patients have 13
It has reached 700,000 and is increasing.

On the other hand, carbohydrates and sugars ingested from the diet are first roughly decomposed by the action of α-amylase contained in saliva and α-amylase secreted from the pancreas. Next, by the action of α-glucosidase, a disaccharide-degrading enzyme such as maltase or sucrase, which cleaves the glucosidic bond present at the non-reducing end, it is finally decomposed into glucose as a monosaccharide and absorbed from the cilia on the mesentery. Is being done. Thereafter, the absorbed glucose causes a temporary rise in blood glucose level, and hyperglycemic symptoms occur. Normally,
With rapid insulin secretion and response to normal insulin, the blood glucose level is restored.

However, in the state where sensitivity to insulin is lowered due to overeating, diabetes, and abnormal lipid metabolism, it takes time to return to the original state, and secretion of a large amount of insulin is promoted. This is a so-called glucose intolerance state. Such symptoms further promote obesity and diabetes.

Further, if hyperglycemia is continued, it leads to arteriosclerosis, renal damage, retinopathy and other serious diabetic complications due to glycation reaction with intravascular proteins. In order to prevent or ameliorate such complications, strict glycemic control and suppression of postprandial hyperglycemia, and
It is considered important to suppress excessive insulin secretion and eliminate the burden on the pancreas. In fact, a disaccharide-degrading enzyme inhibitor has recently been developed as a drug, and its usefulness for diabetes and the like has been recognized. In other words, the onset of diabetes and obesity are also considered to be caused by such a decrease in insulin sensitivity.
It is very important to suppress such postprandial hyperglycemia and excessive insulin secretion.

However, at present, there are few foods that can be ingested daily and prevent or improve them safely. In other words, the fact is that it has a high saccharide-degrading enzyme inhibitory activity and is safe to the human body even when it is ingested daily as a food and has no side effects, and it has not reached the stage of practical use.

[0007]

DISCLOSURE OF THE INVENTION The present invention solves the above-mentioned problems of the prior art, has a high glycolytic enzyme inhibitory activity, and is safe for the human body even when ingested as food and has no side effects. It is intended to provide an enzyme inhibitor. Further, the present invention contains the above-mentioned glycolytic enzyme inhibitor, by effectively suppressing postprandial hyperglycemia, diabetes,
It is also an object to provide foods capable of preventing, treating and improving diseases such as obesity and hyperlipidemia.

[0008]

[Means for Solving the Problems] In order to achieve the above-mentioned object, the present inventors have targeted low-polarity, medium-polarity, high-polarity organic solvent-soluble fractions and water-soluble fractions for about 300 kinds of edible plants. Separated into sex fractions, and in each
We searched for components with α-glucosidase inhibitory activity in the tro system. As a result, surprisingly, Yerba mate (scientific name: Ilex paraguari) has been used as tea for many years.
The present inventors have found that the highly polar organic solvent-soluble fraction of leaves has an excellent glycolytic enzyme inhibitory action, and have completed the present invention based on this finding.

That is, the present invention according to claim 1 provides a glycolytic enzyme inhibitor characterized by containing a mate leaf extract obtained by extracting mate leaves as an active ingredient. .

In the present invention according to claim 2, the extract of yerba mate is an extract easily soluble in at least one of water, a highly polar organic solvent and a medium polar organic solvent. An enzyme inhibitor is provided.

The present invention according to claim 3 provides the glycolytic enzyme inhibitor according to claim 2, wherein the highly polar organic solvent is at least one of ethanol, methanol and butanol.

The present invention according to claim 4 provides the glycolytic enzyme inhibitor according to claim 2, wherein the medium polar organic solvent is ethyl acetate.

The present invention according to claim 5 provides a food containing the glycolytic enzyme inhibitor according to any one of claims 1 to 4.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION The saccharide-degrading enzyme inhibitor of the present invention according to claim 1 is characterized by containing a mate leaf extract obtained by extracting mate leaves as an active ingredient. Mate (Yerba mate, scientific name: Ilex paraguariensis)
It is a kind of plant belonging to the Ilexaceae family, and its leaves are used for drinking as tea, but it has never been known that it has a glycolytic enzyme inhibitory activity.
In addition to raw mate leaves, there are semi-dried products, dried products, and the like, and any of these can be used in the present invention. Mate leaves are usually used after being appropriately crushed or shredded.

Here, the mate leaf extract is preferably an extract which is easily soluble in at least one of water, a highly polar organic solvent and a medium polar organic solvent, as in the present invention according to claim 2. Specific examples of the highly polar organic solvent include alcohols such as ethanol, methanol, butanol, and isopropyl alcohol. Among these, as the highly polar organic solvent, it is preferable to use any one or more of ethanol, methanol and butanol as in the present invention according to claim 3.

On the other hand, examples of the medium polar organic solvent include organic solvents such as ethyl acetate, dichloromethane, acetone and chloroform. Of these, ethyl acetate is preferably used as the medium-polar organic solvent, as in the present invention according to claim 4.

Such a mate leaf extract which is easily soluble in at least one of water, a highly polar organic solvent and a medium polar organic solvent can be obtained, for example, by the following method.

First, the mate leaves are used as they are, or if necessary, crushed or shredded. In this way, as it is, or appropriately crushed or shredded mate leaves,
For example, extraction is performed using at least one or more of the above-mentioned water, highly polar organic solvent, and medium polar organic solvent. Of these, the water extract itself is not always sufficient, so it is desirable to use a highly polar organic solvent-soluble component, especially an ethanol-eluted component, in a column such as HP20, excluding the water-eluted component.

Specifically, for example, 5 to 30 times by weight of water is added to the mate leaves, and the mixture is put in a boiling bath for 5 to 120 minutes.
Preferably it is heated for 20-40 minutes, or at room temperature for 1
Let stand for at least one day to perform extraction. By this operation, it is possible to obtain a mate leaf extract (water extract) that is a water-soluble mate leaf extract.

The water extract is further subjected to partition chromatography using water and a highly polar organic solvent (such as butanol) to obtain a water-soluble mate leaf extract (water-soluble substance) and a highly polar organic solvent. It is possible to obtain a mate leaf extract (soluble in a highly polar organic solvent) that is soluble in water.

On the other hand, by continuously eluting with water and a highly polar organic solvent in a chromatography using a column such as HP20 column, the water-soluble mate leaf extract (water extract by a hydrophobic column) is also obtained. It is possible to obtain a mate leaf extract that is soluble in a high-polar organic solvent (high-polar organic solvent extract using a hydrophobic column). As described above, since the water extract itself is not always sufficient, it is desirable to use a highly polar organic solvent-soluble component, especially an ethanol-eluted component, in a column such as HP20 excluding the water-eluted component.

In addition to the above examples, the highly polar organic solvent is used in an amount of 5 to 30 times, preferably 7 times, the amount of mate leaves.
The mixture is added in an amount of ˜12 times, refluxed for 1 to 4 hours, preferably 2 to 3 hours, and filtered if necessary. This operation can be repeated twice or more. By this operation, it is possible to obtain a mate leaf extract that is soluble in a highly polar organic solvent. Furthermore, the obtained filtrates are combined, dried, and subjected to partition chromatography using water and a medium-polar organic solvent to obtain a water-soluble mate leaf extract (water-soluble fraction) and a medium-polar organic solvent. A soluble mate leaf extract (medium-polar organic solvent-soluble fraction, medium-polar organic solvent extract) can also be obtained. Subsequently, the water-soluble fraction was subjected to partition chromatography with water and a highly polar organic solvent to obtain a mate leaf extract (a highly polar organic solvent extract) soluble in the highly polar organic solvent and a water-soluble mate. Leaf extract (water extract) can also be obtained.

The mate leaf extract thus obtained can be used as it is as a glycolytic enzyme inhibitor, but the residue is further removed by, for example, centrifugation, filtration, pressing or other solid-liquid separation means. If necessary, the product can be formulated as it is, or after being concentrated under reduced pressure, dried under reduced pressure, freeze-dried and the like.

Thus, the target glycolytic enzyme inhibitor of the present invention according to claim 1 can be obtained. The thus-obtained glycolytic enzyme inhibitor of the present invention according to claim 1 has an excellent inhibitory activity against a glycolytic enzyme. Glycolytic enzymes are enzymes that degrade sugars, and include, for example, maltase that decomposes maltose, α-glucosidase such as sucrase that decomposes sucrose, and α-amylase that decomposes starch.

The presence or absence and the degree of the glycolytic enzyme inhibitory activity can be confirmed by a glycolytic enzyme inhibitory test or a sugar tolerance test. Glycolytic enzyme inhibition test, maltose or sucrose which is a substrate of α-glucosidase, starch which is a substrate of α-amylase, is added to the sample, and after reacting by adding a mate leaf extract, glucose released by glucose oxidase method is used. This is a quantitative test. The glucose tolerance test is a test in which a rat is administered with only a substrate such as maltose, sucrose, or starch, or a substrate and a mate leaf extract and the blood glucose level after administration is measured.

The glycolytic enzyme inhibitor of the present invention according to claim 1 does not have an absorption inhibitory action on glucose itself or a glucose lowering action other than the glucose absorption inhibitory action, but has a glycolytic enzyme inhibitory action. Therefore, it is recognized that it inhibits the decomposition of sugar and suppresses the rise in blood sugar.

The glycolytic enzyme inhibitor of the present invention according to claim 1 can be made into various forms by combining it with other optional components. In particular, as in the present invention according to claim 4, it is preferably used as a form of food. That is, the present invention according to claim 5 is the above-mentioned claim 1
The present invention provides a food containing the glycolytic enzyme inhibitor according to any one of 1 to 4.

In the present invention according to claim 5, for example, the water extract as described above can be directly used as a beverage containing a glycolytic enzyme inhibitor, or the extract powder can be directly used as a product. Good, but in order to exert the effect efficiently, preferably an extract containing a large amount of high polar organic solvent soluble components of yerba mate, for example, water-extracted extract is passed through an HP20 column to remove water-eluted components, and then adsorbed components. It is preferable to use the extract obtained by eluting the alcohol with alcohol as a product. Further, this may be made into various forms such as tablets, capsules and soft capsules, and suitable excipients (eg, dextran, oligosaccharides, lactose, etc.) are added to these forms. It may be one.

Further, the food of the present invention according to claim 5 is
As long as it contains the above-mentioned glycolytic enzyme inhibitor, if necessary, a herbal medicine component, a vitamin agent, a stabilizer, a preservative, an antioxidant, a sweetener, a coloring agent, a flavoring agent, a fruit juice, etc. May be There is no particular limitation on the form of the food here, including drinks, bread and biscuits,
Further, it can be used in the form of confectionery. It can also be used as a so-called health food. Further, it can be used as pet food.

The amount of the above-mentioned glycolytic enzyme inhibitor to be mixed in the food of the present invention according to claim 5 is not particularly limited, but is usually 0.1 to 5% by weight, preferably 0.1% by weight in the above-mentioned form. It is about 5 to 1% by weight.

[0031]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to the following examples. In the following, first, the procedure of an in vitro disaccharide-degrading enzyme inhibition test (α-glucosidase inhibition test) and a sugar absorption inhibition test (sugar tolerance test) using animals is shown, and then the preparation of yerba mate extract The procedure of (Production Example 1
4), and the results (Examples 1 to 7) of the various tests and enzyme properties of the prepared yerba mate extract are shown.

[Α-Glucosidase Inhibition Test and Glucose Tolerance Test] I. α-Glucosidase Inhibition Test As α-glucosidase (maltase), rat small intestine acetone powder (manufactured by Sigma) was homogenized with 0.1 M phosphate buffer. , 1% of the precipitate obtained by centrifugation
Dissolve in Triton-X100, centrifuge, then dialyze the supernatant,
Used as crude enzyme. In the control area, maltose (14 mM)
Was used as a substrate, a crude enzyme solution was added, and after 15 minutes of reaction, released glucose was quantified by the glucose oxidase method. In the test substance group (the present invention group), glucose was quantified by reacting in the same manner as in the control group except that the fractions from the mate obtained in Production Examples 1 to 4 were added to the reaction solution. Regarding the inhibition rate, the activity when the test substance was not added was defined as 100, and the amount obtained by subtracting the activity when the test product was added from 100 was shown as the inhibition rate (%).

II. Glucose tolerance test Male Donryu rats (7 weeks old) were used for the experiment after preliminarily breeding for 1 week. After an overnight fast, maltose was added to the control group
2.5 g / kg was orally administered using a sonde, and the mate fraction was administered simultaneously with maltose to the subject group. After administration, blood was collected from the tail vein over time, and the blood glucose level was measured.

Production Example 1 8000 ml of water was added to 1000 g of mate dry leaves, which was extracted in a boiling bath for 30 minutes, filtered, concentrated, freeze-dried, and hot water extract 230 g.
Got

Production Example 2 50 g of the hot water extract obtained in Production Example 1 was subjected to partition chromatography with water and butanol to obtain a water-soluble substance.
37.9 g and butanol soluble matter 12.1 g were obtained.

Production Example 3 The hot water extract obtained in Production Example 1 was subjected to HP20 column chromatography. An HP20 column (trade name: Diaion) in which 50 g of hot water extract was dissolved in water and equilibrated with water
HP20, manufacturer: Mitsubishi Chemical Corporation sampled,
The column was eluted with 5 volumes of water. Then, the solvent was changed to ethanol and the elution was performed in the same amount by 5 times. By this operation, HP2
03.3 column water eluate 33.3g and ethanol eluate 16.7g
Got

Production Example 4 To 200 g of dried mate leaves, 2000 ml of methanol was added and the mixture was refluxed for 3 hours. After filtration, 2000 ml of methanol was added to the residue again and the mixture was refluxed for 3 hours. The filtrates were combined and concentrated to dryness to obtain 41.4 g of methanol extract powder. The methanol extract powder was further subjected to partition chromatography with ethyl acetate and water to separate it into an ethyl acetate-soluble fraction (ethyl acetate-soluble matter) and a water-soluble fraction. The water-soluble fraction was subjected to butanol / water partition chromatography to separate it into a butanol-soluble fraction (butanol-soluble matter) and a water-soluble fraction (water-soluble matter). As a result, 5.5 g, 5.5 g and 15.2 g of ethyl acetate soluble matter, butanol soluble matter and water soluble matter were obtained, respectively.
got g.

Example 1 Using the hot water extract of yerba mate obtained in Production Example 1 as a test substance, an α-glucosidase inhibition test and a sugar load test were carried out by the above-mentioned method, and an α-glucosidase inhibitory action and a sugar load were carried out. The subsequent inhibitory effect on blood glucose elevation was examined. In the α-glucosidase inhibition test, the concentration of the test substance (hot water extract of mate leaf) in the reaction solution was 1000 μg / ml, 100 μ
The concentration was g / ml. In addition, in the glucose tolerance test, the dose of each test substance was 500 mg / kg (body weight) per rat. The results of the glucose tolerance test are shown in FIG.

Regarding the results of the α-glucosidase inhibition test, the inhibition rates were 78.6% and 52.2% at concentrations of 1000 μg / ml and 100 μg / ml in the reaction solution, respectively. Regarding the results of the glucose tolerance test, as is clear from FIG. 1, hot water extract administration
After 30 minutes and 60 minutes, there was a significant decrease (p <0.05) with respect to the control group (Control). From these facts, it was revealed that the hot water extract of yerba mate obtained in Production Example 1 inhibits α-glucosidase (maltase) and suppresses an increase in blood glucose after glucose loading.

Example 2 The butanol-soluble substance (BuOH fraction) and the water-soluble substance (water fraction) obtained in Production Example 2 were used as test substances, and the α-glucosidase inhibition test and the sugar were conducted by the above-mentioned method. A stress test was conducted to examine the α-glucosidase inhibitory action and the blood glucose elevation inhibitory action after glucose loading. In the α-glucosidase inhibition test, the concentrations of the test substance in the reaction solution were 1000 μg / ml and 100 μg / ml as in Example 1. On the other hand, the dose of each test substance in the glucose tolerance test was determined as in Example 1.
In the same manner as described above, the dose was 500 mg / kg (body weight) per rat. The results of the glucose tolerance test are shown in FIG.

Regarding the results of the α-glucosidase inhibition test, the inhibition rates of the butanol soluble matter (BuOH fraction) were 92.3% and 74.7% at the concentrations of 1000 μg / ml and 100 μg / ml in the reaction solution, respectively.
The inhibition rate of water-soluble matter (water fraction) was 81.6% and 51.
It was 4%. The results of the glucose tolerance test are shown in FIG.
As is clear from the results, the butanol-soluble substance (BuOH fraction) -administered group was treated with the control group (Control
) Was significantly decreased (p <0.01). On the other hand, the water-soluble matter (water fraction) -administered group also showed a decrease compared to the control group (Control), and a slight decrease (p <0.05) especially 30 minutes after administration
It has been shown. From these facts, the butanol-soluble product (BuOH fraction) and the water-soluble product (water fraction) obtained in Production Example 2 inhibited α-glucosidase (maltase),
It became clear that the rise in blood glucose after glucose load was suppressed.

Example 3 The hot water extract of yerba mate obtained in Production Example 3 was applied to the HP20 column.
Water eluate (HP20 / H ₂O) and ethanol eluate (HP20 / E
tOH) and α are measured by the above method.
-Glucosidase inhibition test and glucose tolerance test
Inhibitory effect of lucosidase and suppression of blood glucose increase after glucose loading
I examined it. Odor of α-glucosidase inhibition test
The concentration of the test substance in the reaction solution was the same as in Example 1.
The concentrations were 1000 μg / ml and 100 μg / ml. For glucose tolerance test
The dose of each test substance was the same as in Example 1 for rats 1.
It was set to 500 mg / kg (body weight) per individual. Conclusion of glucose tolerance test
The results are shown in FIG.

Regarding the results of the α-glucosidase inhibition test, the inhibition rates of the water eluate (HP20 / H ₂ O) were 35.2% and 4.4% at 1000 μg / ml and 100 μg / ml concentrations in the reaction solution, respectively.
Inhibition rate of ethanol eluate (HP20 / EtOH) is 91.3%, 69.1
%Met. The results of the glucose tolerance test are shown in FIG.
As is clear from the results, the ethanol eluate (HP20 / EtOH) -administered group was treated with the control group (Contrr
ol) significantly decreased (p <0.01),
The water-soluble matter (HP20 / H ₂ O) -administered group showed a slight decrease (p <0.05) only after 30 minutes. From these things, Production Example 3
Water-eluted product (HP20 / H ₂ O) and ethanol-eluted product (HP20 / EtOH) obtained by the HP20 column inhibit α-glucosidase (maltase) and suppress blood glucose increase after glucose loading. Became clear.

Example 4 The ethyl acetate-soluble matter (ethyl acetate fraction), butanol-soluble matter (BuOH fraction) and water-soluble matter (water fraction) obtained in Production Example 4 were respectively compared as a comparative sample. As the above, a glucose tolerance test was conducted by the above-mentioned method, and the inhibitory effect on blood glucose increase after glucose tolerance was examined. The dose of each test substance is 500 per rat.
It was set to mg / kg (body weight). The results are shown in Fig. 4.

As is clear from FIG. 4, the ethyl acetate soluble matter eluate (ethyl acetate fraction) administration group, butanol soluble matter (BuOH
Fraction) administration group, water-soluble matter (water fraction) administration group,
Control group (Control) 30 and 60 minutes after administration
Was significantly decreased. In particular, butanol soluble matter (Bu
The OH fraction) -administered group already showed a slight decrease after 15 minutes and 30 minutes (p <0.05), and showed a significant suppression after 60 minutes (p <0.05).
From <0.01), it is clear that the blood glucose level is most strongly lowered. From these facts, the blood glucose after sugar loading was determined by the ethyl acetate-soluble matter (ethyl acetate fraction), butanol-soluble matter (BuOH fraction) and water-soluble matter (water fraction) obtained in Production Example 4. It was revealed that the increase was suppressed, and in particular, the butanol-soluble product (BuOH fraction) had a strong hypoglycemic effect.

Example 5 With respect to the butanol-soluble product obtained in Production Example 4, the mode of inhibition of α-glucosidase (maltase) was examined. That is, using maltose as a substrate, Michaelis constant
With respect to the substrate concentration S near the Km value, the reaction rate v for each substrate concentration S was determined under the condition that the concentration of butanol-soluble matter was constant. 1 / s (% ^-1 ) on the horizontal axis, 1 / v (mg-glucose)
se / ml · min) is plotted on the vertical axis (Lineweaver-Bu
rk plot). Results are shown in FIG.

From FIG. 5, it can be seen that the slope of the plot increases as the concentration of the butanol-soluble substance increases, and that the straight lines of each plot do not intersect on the vertical axis but intersect on the horizontal axis. From this, it was shown that the inhibition form of the butanol soluble substance with respect to α-glucosidase is a non-antagonistic type.

Example 6 With respect to the butanol-soluble product obtained in Production Example 4, α-amylase inhibitory action and α-glucosidase (maltase, sucrase) inhibitory action on starch, sucrose and maltose, and inhibition of blood glucose increase after glucose loading. The effect was examined. That is, using the butanol-soluble product obtained in Production Example 4 as a test substance, an α-glucosidase inhibition test was conducted by the above-mentioned method, and the α-glucosidase (maltase) inhibitory action was examined. Further, in this α-glucosidase inhibition test, an α-amylase inhibition test using starch (manufactured by Wako Pure Chemical Industries, Ltd.) in place of maltose was conducted to examine the α-amylase inhibitory action. further,
In this α-glucosidase inhibition test, sucrose (manufactured by Wako Pure Chemical Industries, Ltd.) was used instead of maltose.
-A glucosidase (sucrase) inhibition test was performed, and α-
A glucosidase (sucrase) inhibitory effect was examined. In these inhibition tests, the concentration of the test substance (hot water extract of yerba mate) in the reaction solution was 1000 μg / ml, 100
The concentration was μg / ml.

Further, the butanol-soluble substance obtained in Production Example 4 was used as a test substance, and a glucose tolerance test was conducted using starch and sucrose in addition to maltose as a substrate, and the inhibitory effect on blood glucose increase after the glucose tolerance was examined. investigated. The dose of the butanol soluble substance, which is the test substance, is 250 mg per rat.
/ Kg (body weight) and 500 mg / kg (body weight). The results of the glucose tolerance test with maltose, starch and sucrose as the substrates
Shown in Figures 6, 7 and 8, respectively.

As a result of the α-glucosidase (maltase) inhibition test, the reaction solution was tested at concentrations of 1000 μg / ml and 100 μg / ml.
The inhibitory activities were 81.1% and 54.8%. In addition, as a result of α-amylase inhibition test, it was found that 1000 μg / ml, 100 μg / m in the reaction solution.
At 1 concentration, the inhibitory activities were 72.3% and 50.4%, respectively.
Furthermore, as a result of the α-glucosidase (sucrase) inhibition test, the inhibitory activities were 57.4% and 28.1% at the concentrations of 1000 μg / ml and 100 μg / ml in the reaction solution. Therefore, it was revealed that the butanol-soluble product obtained in Production Example 4 exhibited a strong inhibitory action on any of maltase, α-amylase and sucrase. In particular, since the inhibitory activity was higher when maltose was used as the substrate than when starch or sucrose was used as the substrate, the butanol-soluble product showed a slightly higher maltase activity than the α-amylase and sucrase activity inhibitory activity. It was shown that the inhibitory effect was strong.

Further, as is clear from FIGS. 6 to 8, the results of the glucose tolerance test also showed that the glucose-suppressing action after glucose loading was exhibited in both cases of maltose loading, starch and sucrose loading. It was

That is, as is clear from FIG. 6, when maltose was loaded, the butanol-soluble substance obtained in Production Example 4 was 250 mg / kg (rat) at 30 minutes and 60 minutes after administration. Both body weight) and 500 mg / kg (body weight) were significantly lower than the control group (Control) (p <0.01).

As is clear from FIG. 7, when starch was loaded, the butanol soluble product obtained in Production Example 4 was administered at a dose of 250 mg / kg (body weight) per rat for 60 minutes after administration. After that, a slight decrease was shown with respect to the control group (Control) (p <0.05). Similarly, when loaded with starch, the butanol soluble matter obtained in Production Example 4 was
At a dose of 500 mg / kg (body weight) per rat, 15 minutes, 30 minutes, and 60 minutes after administration, it was significantly decreased (p <0.01) with respect to the control group (Control).

Further, as is clear from FIG. 8, when sucrose was loaded, the butanol-soluble product obtained in Production Example 4 was administered at a dose of 250 mg / kg (body weight) per rat for 30 minutes after administration. And 120 minutes later, the control group (Control
) Showed a slight decrease (p <0.05). Similarly, when loaded with starch, the butanol-soluble matter obtained in Production Example 4 was administered at a dose of 500 mg / kg (body weight) per rat at 15 minutes, 30 minutes, 60 minutes after administration, respectively. , Significantly lower than the control (Control) (p <
0.01).

Example 7 The butanol-soluble substance obtained in Production Example 4 was subjected to a glucose tolerance test using glucose as a substrate instead of maltose, and the inhibitory effect on the blood glucose increase after the glucose tolerance was examined. The dose of the butanol-soluble substance to be tested is 250 mg.
/ Kg. As a result, there was no effect on the rise in blood glucose after glucose load. Further, when only the butanol-soluble substance obtained in Production Example 4 was orally administered, no influence was observed on the blood glucose level. From these facts, the effect of suppressing the increase in blood sugar after sugar loading by the mate leaf extract is not due to the effect of lowering blood sugar other than the effect of suppressing absorption of glucose itself or the effect of suppressing sugar absorption, but the effect of inhibiting carbohydrate degradation by the action of glycolytic enzyme. It was considered that the decomposition was inhibited.

[0056]

EFFECTS OF THE INVENTION The glycolytic enzyme inhibitor of the present invention according to claim 1 exhibits a strong inhibitory action on various glycolytic enzymes including α-amylase and α-glucosidases such as maltase and sucrase. . The glycolytic enzyme inhibitor of the present invention according to claim 1 has a strong inhibitory effect on maltase that decomposes maltose, and suppresses the decomposition of starch itself and the disaccharide degrading enzyme inhibitory effect in the final absorption process. It is considered that by suppressing postprandial hyperglycemia, it has a preventive and therapeutic effect on not only diabetes but also lifestyle-related diseases such as obesity and hyperlipidemia.

Since the food of the present invention according to claim 2 contains the glycolytic enzyme inhibitor of the present invention according to claim 1, it has an excellent glycolytic enzyme inhibitory action. And since it is a material that has been eaten for many years, it is safe and is extremely excellent for daily intake and use. Further, since it has a strong activity by hot water treatment, it can be used by adding it to various foods, processed foods, pharmaceuticals and the like. Therefore, it can be effectively used as a food for preventing or treating not only diabetes but also lifestyle-related diseases such as obesity and hyperlipidemia.

[Brief description of drawings]

FIG. 1 is a graph showing the results of a sugar tolerance test of a hot water extract of yerba mate in Example 1.

FIG. 2 is a graph showing the results of a glucose tolerance test of a butanol-soluble matter (BuOH fraction) and a water-soluble matter (water fraction) of yerba mate in Example 2.

FIG. 3 A water eluate (HP20 / H ₂ O) and an ethanol eluate (HP20 / EtO) of the mate leaf in Example 3 by HP20 column.
It is a graph which shows the result of the glucose tolerance test of H).

[Fig. 4] Fig. 4 shows the results of a sugar load test of ethyl acetate-soluble matter (ethyl acetate fraction), butanol-soluble matter (BuOH fraction), and water-soluble matter (water fraction) of mate leaves in Example 4. It is a graph.

FIG. 5 is a graph showing the results of a Lineweaver-Burk plot of a butanol-soluble material of yerba mate in Example 5.

FIG. 6 is a graph showing the results of a sugar tolerance test in the case where the substrate was maltose with respect to the butanol-soluble matter of mate leaves in Example 6.

FIG. 7 is a graph showing the results of a sugar tolerance test in the case where the substrate is starch for the butanol soluble matter of mate leaves in Example 6.

FIG. 8 is a graph showing the results of a sugar tolerance test in a case where the substrate was sucrose with respect to the butanol soluble matter of mate leaves in Example 6.

─────────────────────────────────────────────────── ─── Continued Front Page (51) Int.Cl. ⁷ Identification Code FI Theme Coat (Reference) A61P 43/00 111 A61P 43/00 111 C12N 9/99 C12N 9/99 (72) Inventor Tetsuko Teramoto Okayama Prefecture Kurashiki City Chayamachi 353-56 F Term (reference) 4B018 MD61 ME03 ME04 ME14 MF01 4C088 AB12 AC05 BA08 BA09 BA10 CA05 CA06 CA07 MA52 MA70 NA14 ZA70 ZC20 ZC33 ZC35

Claims (5)

[Claims] 1. A glycolytic enzyme inhibitor, which comprises a mate leaf extract obtained by extracting mate leaves as an active ingredient. 2. The glycolytic enzyme inhibitor according to claim 1, wherein the yerba mate extract is an extract which is easily soluble in at least one of water, a highly polar organic solvent and a medium polar organic solvent. 3. The glycolytic enzyme inhibitor according to claim 2, wherein the highly polar organic solvent is at least one of ethanol, methanol and butanol. 4. The glycolytic enzyme inhibitor according to claim 2, wherein the medium polar organic solvent is ethyl acetate. 5. A food containing the glycolytic enzyme inhibitor according to claim 1.