JP7429169B2 - Method for producing adzuki bean-derived glycolytic enzyme inhibitor - Google Patents

Description

Application of Article 30, Paragraph 2 of the Patent Act Publication of research on an adzuki bean-derived sugar metabolic enzyme inhibitor and its manufacturing method at the 36th Mie University regular press conference (October 31, 2020)

Application of Article 30, Paragraph 2 of the Patent Act Journal of Food Science Volume 84, Issue 11 Pages 3172-3178 published research on an adzuki bean-derived sugar metabolic enzyme inhibitor and its manufacturing method (October 15, 2021)

Application of Article 30, Paragraph 2 of the Patent Act Research on an adzuki bean-derived sugar metabolic enzyme inhibitor and its manufacturing method was disclosed at the 2019 Chubu Branch Conference of the Japan Society for Food Science and Technology (December 14, 2020)

The present invention relates to a method for producing a glycolytic enzyme inhibitor derived from azuki bean, and particularly to a method for producing an extract derived from azuki bean that effectively inhibits the activity of a glycolytic enzyme involved in sugar absorption.

Adzuki bean (Vigna angularis) is mainly processed to soften by heating by boiling, and is used as an ingredient for dishes such as sekihan, or as an ingredient for confectionery such as ``Oguraan'' with sugar added. It has been eaten since ancient times. It is known that the skin of azuki beans contains polyphenols, saponins, and even glycosides bonded to sugar molecules. These ingredients have been reported to have antiallergic activity, hypoglycemic activity, cell adhesion inhibition, hyperlipidemia prevention, bone metabolism regulation, and other effects through analysis of adzuki bean broth, or hot water extract (patented). (See Document 1, etc., Non-Patent Documents 1, 2, 3, 4, 5, etc.).

Among natural products such as the components contained in azuki beans, research is being carried out particularly on the components and effects related to sugar metabolism. In particular, abnormalities in sugar metabolism make it difficult to control blood sugar levels, which is a major cause of diabetes. When diabetes becomes severe, it causes blindness and peripheral necrosis. Diabetes is a type of lifestyle-related disease, and there is concern that it may develop due to irregular eating habits and lifestyle habits. If this is the case, it may be possible to suppress the onset of the disease by taking measures based on daily eating habits and lifestyle habits.

Based on this point, findings suggesting a relationship between polyphenols contained in natural compounds derived from foods and sugar metabolism have been reported (see Non-Patent Documents 6 and 7, etc.). However, the specific relationship between sugar metabolism and the contained components has not been elucidated. Subsequently, a relationship between polyphenol catechins and sugar metabolic enzymes has been reported (see Non-Patent Document 8, etc.).

Patent No. 6023398 Patent No. 6031328

T. Itoh et al. , Biosci. Biotechnol. Biochem. , 68(12), 2421-2426 (2004) T. Itoh et al. , Biosci. Biotechnol. Biochem. , 69(3), 448-454 (2005) T. Itoh et al. , Nutrition, 25, 134-141 (2009) T. Itoh et al. , Nutrition, 25, 318-321 (2009) T. Itoh et al. , Phytother. Res. , 26(7), 1003-1011 (2012) Michiyuki Kojima et al., Journal of the Japanese Society of Food Science and Technology, Volume 54, No. 1, 50-53 (2007) Yusuke Saito et al., Journal of the Japanese Society of Food Science and Technology, Volume 54, No. 12, 563-567 (2007) Tae Jung Ha et al. , Food Chemistry, 266, 483-489 (2018)

Based on a series of circumstances, the inventor conducted extensive research on the components contained in the seeds of azuki beans, which have been eaten regularly since ancient times, and found that catechins in azuki beans are related to sugar metabolism (sugar decomposition) depending on the type. We obtained the knowledge that it acts on enzymes. In other words, this suggests that the catechins in azuki beans may be effective in improving and preventing sugar metabolism and hyperglycemia.

The present invention has been made in view of the above points, and provides a method for producing a glycolytic enzyme inhibitor that utilizes components contained in adzuki bean seeds.

The following will achieve the above objectives:
(1) A soaking process in which the azuki beans are immersed in water to obtain a soaking liquid, a soaking liquid recovery process in which the soaking liquid and the hydrated adzuki beans are separated and the soaking liquid is recovered, and a heating process in which the collected soaking liquid is heated. Adzuki bean-derived glycolytic enzyme containing (+)-catechin 7-O-β-D-glucopyranoside and (+)-epicatechin 7-O-β-D-glucopyranoside and inhibiting the enzymatic activity of α-glucosidase. Method for producing inhibitors.

(2) The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to (1) above, wherein the soaking time in the soaking step is 8 to 12 hours.

(3) The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to (1) or (2) above, wherein the amount of water used for soaking the adzuki beans in the soaking step is from the same amount to twice the weight of the adzuki beans. .

(4) The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to any one of (1) to (3) above, wherein the temperature in the heating step is 90 to 100°C.

(5) The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to any one of (1) to (4) above, wherein the heating time in the heating step is 60 to 120 minutes.

(6) The soaking time in the soaking step is 8 to 12 hours, the amount of water used for soaking the adzuki beans in the soaking step is the same amount to twice the weight of the adzuki beans, and the temperature in the heating step is is 90 to 100°C, and the heating time in the heating step is 60 to 120 minutes.

The present invention includes a soaking process of soaking adzuki beans in water to obtain a soaking liquid, a soaking liquid recovery process of separating the water-containing adzuki beans from the soaking liquid and recovering the soaking liquid, and a heating process of heating the collected soaking liquid. and contains (+)-catechin 7-O-β-D-glucopyranoside and (+)-epicatechin 7-O-β-D-glucopyranoside, which inhibits α-glucosidase enzyme activity. This is a method for producing an enzyme inhibitor.

Therefore, an adzuki bean-derived glycolytic enzyme inhibitor containing (+)-catechin 7-O-β-D-glucopyranoside and (+)-epicatechin 7-O-β-D-glucopyranoside can be produced.

FIG. 1 is a schematic process diagram illustrating the preparation of an adzuki bean-derived glycolytic enzyme inhibitor. It is a chart of peaks of high performance liquid chromatography of an extract. This is NMR spectrum data of C7G. This is NMR spectrum data of E7G. This is NMR spectrum data of catechin. It is a chart of high performance liquid chromatography peaks of C7G and E7G.

The preparation process of the adzuki bean-derived glycolytic enzyme inhibitor (adzuki bean-derived extract for regulating glycolysis ) of the embodiment will be explained using the schematic process diagram of FIG. 1. First, untreated adzuki beans (Adzuki beans: Vigna angularis) are appropriately washed with water and then soaked in water (ultra-pure water in the embodiment) to obtain a soaking liquid ("soaking step"). The soaking time is 8 to 12 hours (8 to 12 hours), taking into account seasonal temperature fluctuations. The amount of water used for soaking the adzuki beans is the same amount to twice the weight of the adzuki beans (same amount to twice the amount) .

The immersion liquid produced by immersion in water has a light reddish-brown color. The soaking liquid and the water-containing azuki beans are separated (drained) and only the soaking liquid is recovered. Normally, the azuki bean soaking liquid (water soaking liquid) is discarded to remove unpleasant tastes such as astringency. The hydrated adzuki bean is then heated, an appropriate amount of sugar is added, and it is processed into Ogura bean paste, a type of confectionery ingredient. The noteworthy point of the adzuki bean-derived glycolytic enzyme inhibitor of the embodiment is that it uses the adzuki bean soaking liquid, which is generally discarded during the production of adzuki bean paste, as a starting point.

The addition of 70 to 90% by weight methanol aqueous solution to the immersion liquid is due to the difference in detection components in the Examples described below. In the experiment in the example, a concentration gradient of methanol (initially 0 to 100 gradually) was added to the immersion liquid, and the mixture was subjected to HPLC (high performance liquid chromatography). Then, the methanol concentration at the time of peak detection was specified.

Furthermore, prior to the separation step, the immersion liquid is heated (“heating step”). The heating of the immersion liquid is at boiling temperature, which is between 90 and 100°C (90-100°C) . The heating time is 60 to 120 minutes (60-120 minutes) .

The methanol supernatant produced by the separation using an aqueous methanol solution is dried to obtain a dried product (“drying step”). By drying, a powder of the target component eluted in the immersion liquid can be obtained. In this way, two systems are prepared: a dry solid product (a) without a heating process and a dry solid product (b) with a heating process.

The dried product (a) and the dried product (b) prepared from the azuki beans soaked in water have in common the “(+)-catechin 7-O-β-D-glucopyranoside” of formula (1). Contains. Hereinafter, the catechin will be referred to as "C7G". Catechin with the structure C7G has a plant-derived composition, and its presence in a water soaked adzuki bean solution was also confirmed.

In addition, the dry product (b) with heating step contains "(+)-epicatechin 7-O-β-D-glucopyranoside" of formula (2). Hereinafter, the epicatechin will be referred to as "E7G". The presence of epicatechin in E7G was confirmed by HPLC peak position analysis in Examples described below. C7G and E7G are isomers, and it is thought that when C7G is heated, a part of its structure changes, resulting in E7G.

As shown in the examples, the dried products (dried product (a) and dried product (b)) prepared from the water-soaked adzuki beans based on a series of steps should exhibit the activity of inhibiting glycolytic enzymes. was found. Therefore, on the basis of the presence of glycolytic enzyme inhibitory activity, it can be said that the dried adzuki bean product derived from soaking in water functions as an adzuki bean-derived glycolytic enzyme inhibitor.

The mechanism of action of sugar metabolism is roughly as follows. Carbohydrates (so-called starches and carbohydrates) that humans ingest orally through meals are first broken down into disaccharides by α-amylase in saliva and pancreatic juice. Thereafter, the disaccharides are broken down into monosaccharides by α-glucosidase secreted from the brush border of small intestinal mucosal epithelial cells, and then absorbed from the small intestine. After absorption of sugars, there is a sudden rise in blood sugar levels, which can lead to insulin secretion and a sudden drop in blood sugar levels, which can lead to chronic diseases such as excessive sugar accumulation within cells.

Therefore, by inhibiting the enzymatic activity of α-amylase or α-glucosidase involved in the decomposition of carbohydrates, the production of monosaccharides, which are end products, may be reduced, and a sudden rise in blood sugar levels may be suppressed. Currently, Acarbose, Voglibose, Miglitol, and the like are prescribed as therapeutic drugs for type 2 diabetes. Therefore, based on the findings in the Examples that the adzuki bean-derived glycolytic enzyme inhibitor inhibits enzyme activity to the same extent as these therapeutic and preventive drugs, it is effective in preventing disorders of sugar metabolism such as diabetes.

It has been found that the adzuki bean-derived glycolytic enzyme inhibitor has a higher inhibitory effect on the enzymatic activity of α-glucosidase than existing therapeutic drugs for diabetes. Furthermore, catechins such as C7G catechin and E7G epicatechin contained in the adzuki bean-derived glycolytic enzyme inhibitor inhibit the enzymatic activities of α-amylase and α-glucosidase. In particular, E7G epicatechin acts on α-amylase and α-glucosidase, and is highly effective in inhibiting the enzymatic activity of α-glucosidase.

This effect is new knowledge. Previously, it has been reported that there is a certain degree of correlation between the various pharmacological activities of polyphenols, glycosides, etc. of catechins. However, it has become clear that the effects of each type of enzyme vary due to differences in the structure of catechins. Therefore, adzuki bean-derived glycolytic enzyme inhibitors are extremely promising for the purpose of replacing or supplementing existing therapeutic drugs for type 2 diabetes. In addition, it is also expected to be applied to improving carbohydrate-derived obesity.

When the sugar metabolism regulator derived from azuki bean (extract derived from azuki fruit for regulating sugar metabolism) is used as a prophylactic or therapeutic agent for improving glucose metabolism disorders and diabetes, the dosage form for oral administration is tablets, capsules, powders, and granules. , liquid preparation, etc. as appropriate. Further, it can also be used as an injection, suppository, drip, spray, nasal spray, etc. In the listed drugs, the adzuki bean-derived sugar metabolism regulator may be used alone or in combination with other drugs.

The adzuki bean-derived sugar metabolism regulator of the present invention is a natural product and is an extract derived from azuki bean, which has been eaten for a long time. For this reason, adzuki bean-derived sugar metabolism regulators are compatible with commonly consumed foods. Therefore, it is possible to prepare an adzuki bean-derived sugar metabolism regulator-containing food to which the adzuki bean-derived sugar metabolism regulator is newly added.

Foods to which the azuki-derived sugar metabolism regulator (sugar metabolism-regulating azuki bean-derived extract) can be added include mizu-yokan, yokan, Ogura bean paste, dorayaki, azuki mousse, ice cream, candy, and caramel. , custard pudding, coffee jelly, gummies, Bavarois, marshmallows, castella, pancakes, cookies, fruit juice, carbonated drinks, etc. Further examples include tablets that serve as supplements.

When manufacturing the listed foods containing the adzuki bean-derived glycolysis regulator, the content of the adzuki bean-derived glycolysis regulator is determined based on the taste of the food itself, the change in taste after addition, other added ingredients, and the amount consumed per serving. , consumption frequency, seasonality, sales format, consumer demographics including age and gender, etc. are comprehensively considered. The amount of the adzuki bean-derived sugar decomposition regulator added to the food is arbitrary, and may be, for example, 0.01% to 50% by weight (0.01% to 50% by weight) .

[Extraction of Azuki-derived components]
Adzuki beans (variety: Erimoshozu) from Hokkaido were used as the raw material Azuki (Vigna angularis). A predetermined amount of azuki beans was lightly washed with water, soaked in ultrapure water at room temperature (approximately 15° C.) of the same weight as the azuki beans, and left to stand for 10 hours. After soaking, the soaking liquid (supernatant) and the water-containing azuki beans were separated. The immersion liquid was divided into two types, and one of the immersion liquids was left as is. The other soaking liquid was boiled for 90 minutes and cooled to room temperature. In this way, two types of soaking liquid were obtained: a soaking liquid made by only soaking the azuki beans in water (hereinafter referred to as "supernatant (1)") and a heated soaking liquid (hereinafter referred to as "supernatant (2)"). Ta. Both supernatant (1) and supernatant (2) were light purple-brown in color.

[HPLC analysis]
An ODS column (manufactured by Nacalai Tesque Co., Ltd., COSMOSIL 5C18-AR, 4.6×150 mm) was used. The ODS column was averaged through methanol prior to analysis. The detection wavelength was 210 nm.

When supernatant (1) or supernatant (2) was inserted into HPLC (injection), a methanol aqueous solution with a concentration gradient of water (ultrapure water) and methanol was used as the mobile phase (flow rate 1 mL/min). CDS Lite (manufactured by L.A.Soft) was used to control the concentration gradient of the mobile phase, and recording was performed for 80 minutes.

The conditions for the mobile phase gradient program were as follows.
0 to 15 minutes: Maintain 100% ultrapure water 15 to 45 minutes: Gradually increase concentration from 100% ultrapure water to 100% methanol 45 to 60 minutes: Maintain 100% methanol 60 to 75 minutes: Gradual concentration drop from 100% methanol to 100% ultrapure water The amount of sample inserted was adjusted to 2 μg (concentration 0.2 mg/mL, volume 10 μL) to 10 μg (concentration 1 mg/mL, volume 10 μL).

FIG. 2 is an HPLC peak chart. The vertical axis is the detection potential (mV), and the horizontal axis is the retention time (min). The "upper" chart is the HPLC detection result of supernatant (1), and the "lower" chart is the HPLC detection result of supernatant (2). Peak I and peak II were detected at the retention times in the upper row. . Then, peak I, peak II, and peak III were detected at the lower retention times.

For each of the components detected in peaks I and II of the upper supernatant (1) and the components detected in peaks I, II, and III of the lower supernatant (2), depending on the position of the corresponding peak. Isolated and dried.

[Measurement of glycolytic enzyme inhibitory effect]
First, for both the upper supernatant (1) (water immersion) and the lower supernatant (2) (water immersion and heating) samples, two types related to saccharide degradation were analyzed as a whole (including each component). The inhibitory effect on enzyme activity was determined. α-amylase and α-glucosidase were used as enzymes.

- α-Glucosidase activity inhibition test For the α-glucosidase activity inhibition test, a 96-well plate was used, and yeast-derived α-glucosidase was used. α-Glucosidase was prepared at 0.2 U/mL using 0.1 M phosphate buffer (pH 6.7). The enzyme solution was injected at 50 μL/well into the test group, and the phosphate buffer solution was injected at 50 μL/well into the control group. 50 μL of a sample solution prepared at 0.001 to 10 mg/mL was added to each well as a sample. After pre-incubation at 37°C for 10 minutes, 20 μL of substrate solution PNPG (p-nitrophenyl-β-D-glucoside) solution was added to each well, followed by incubation at 37°C for 30 minutes. Immediately after the incubation, 120 μL of 0.6 M sodium bicarbonate solution was added per well to stop the enzyme reaction. Immediately after the reaction was stopped, the absorbance at 405 nm was measured using a plate reader (CHROMATE MODEL 4300, manufactured by AWARENESS TECHNOLOGY Inc.), and the activity was calculated. Note that the following formula was used for _IC50 . The results are shown in Table 1.

IC ₅₀ = 10 [{log (A/B)・(50-C)・1/(D-C)}
+log(B)]
In the formula, A: the higher concentration of the two points between 50%
B: Lower concentration of the two points between 50%
C: inhibition rate in B
D: inhibition rate in A

- α-Amylase activity inhibition test For the α-amylase activity inhibition test, a 96-well plate was used, and α-amylase derived from porcine pancreas was used. α-amylase was prepared with 0.1M phosphate buffer (pH 6.7) containing 60mM sodium chloride at 0.6U/mL. 50 μL/well of the enzyme solution was injected into the test group, and 50 μL/well of the aforementioned phosphate buffer containing sodium chloride was injected into the control group. 50 μL of a sample solution prepared at 0.001 to 10 mg/mL was added to each well as a sample. After pre-incubation at 37°C for 10 minutes, 20 μL of substrate solution CNP-G (2-chloro-4-nitrophenyl-α-D-maltotrioside) solution was added per well, and incubated at 37°C for 30 minutes. Incubated for 1 minute. Immediately after the incubation, 120 μL of 0.6 M sodium bicarbonate solution was added per well to stop the enzyme reaction. Immediately after the reaction was stopped, the absorbance at 405 nm was measured using a plate reader (CHROMATE MODEL 4300, manufactured by AWARENESS TECHNOLOGY Inc.), and the activity was calculated. For IC ₅₀ , the same formula as in the α-glucosidase activity inhibition test described above was used. The results are shown in Table 2.

[Considerations on measurement of glycolytic enzyme inhibitory effect]
From the results of the α-glucosidase activity inhibition test in Table 1 and the α-amylase activity inhibition test in Table 2, supernatant (1) (water immersion) and supernatant (2) (water immersion and heating ) exhibited inhibitory activity in a concentration-dependent manner. Therefore, it was confirmed that the components eluted in the water soaked adzuki beans inhibited the reaction of two types of enzymes involved in saccharide decomposition .

[Structure determination of eluted components]
In FIG. 2, the components detected in peaks I and II of the upper supernatant (1) and the components detected in peaks I, II, and III of the lower supernatant (2) were analyzed by ¹ H-NMR. The structure was determined using In addition, the molecular weight was determined using LC-MS. In determining the structure by ¹ H-NMR, reference was made to the spectral data of Hori et al., Journal of the Japanese Society of Nutrition and Food Science, Vol. 62, No. 1, 3-11 (2009). The spectral data in Table 3 was used for structural assignment.

As a result of the structure determination, the structure of the component of peak II was determined as Compound I "(+)-catechin 7-O-β-D-glucopyranoside (C7G), MW: 452". The NMR spectrum of C7G is shown in FIG.
The structure of the component of peak III was determined as Compound II "(+)-epicatechin 7-O-β-D-glucopyranoside (E7G), MW: 452". The NMR spectrum of E7G is shown in FIG.
The structure of the component of peak I was determined as Compound III "(+)-catechin, MW: 290". The NMR spectrum of catechin is shown in FIG.

[Measurement of glycolytic enzyme inhibitory effect of C7G and E7G]
It was confirmed that the water extract of azuki beans contained C7G in addition to catechin, and further contained E7G after heating. Therefore, the inhibitory activity of glycolytic enzymes by C7G and E7G alone was measured. The measurement method was the same as the α-glucosidase activity inhibition test and α-amylase activity inhibition test described above. The results of the α-glucosidase activity inhibition test are shown in Table 4, and the results of the α-amylase activity inhibition test are shown in Table 5.

[Considerations on measuring the glycolytic enzyme inhibitory effect of C7G and E7G]
It has been revealed that C7G and E7G each exhibit inhibitory activity against both α-amylase and α-glucosidase, which are involved in glycolysis , even when used alone. Further, although C7G and E7G are similarly concentration dependent, E7G tends to have a higher ability to inhibit enzyme activity than C7G in biochemical tests.

[Generation factors of E7G]
From the HPLC retention time chart in FIG. 2 above, the difference between the upper supernatant (1) and the lower supernatant (2) is that they are only immersed in water or are heated while immersed in water. Here, C7G and E7G have the same molecular weight and are in the relationship of isomers. Therefore, it was confirmed whether E7G is generated from C7G by heating C7G.

The fractionated C7G was dissolved in ultrapure water (concentration 0.05 mg/mL) and heated at boiling temperature for 90 minutes. C7G alone and two types of C7G after heating were detected by HPLC. An ODS column (manufactured by Nacalai Tesque Co., Ltd., COSMOSIL 5C18-AR, 4.6×150 mm) was used. The ODS column was averaged through methanol prior to analysis. The detection wavelength was 210 nm.

When inserting C7G alone and C7G into two types of HPLC after heating (injection volume: 10 μL), a methanol aqueous solution of 20% water (ultra pure water) and 20% methanol was used as the mobile phase (flow rate 1 mL/ min). The peak of HPLC retention time was shown in the chart of FIG.

The upper row is the peak of a solution containing only C7G. A C7G peak exists around the retention time of 3 minutes. The lower row shows the peak of the heated solution of C7G. Along with the C7G peak around the 3 minute retention time, there is another peak around the 6 minute retention time. As a result of comparing this peak with isolated E7G, it was found to be E7G.

From the HPLC results, it was confirmed that C7G changed to E7G upon heating. Furthermore, a calibration curve was created between the heating of C7G and the generation of E7G, and the amount of change from C7G to E7G was investigated. Judging from the calibration curve, it was found that about 20% of C7G was converted to E7G by heating.

[Summary of glycolytic enzyme inhibitory effects]
Extracts obtained by soaking azuki beans in water include supernatant (1) (dry product (a)), supernatant (2) (dry product (b)), C7G, E7G, and existing type 2 The 50% inhibitory concentration (IC ₅₀ ) for both α-amylase and α-glucosidase enzymes was determined for each of Acarbose as a therapeutic drug for diabetes. The quality of the azuki extract components was compared using the existing drug acarbose as a control group. The results are shown in Table 6.

Acarbose exhibits very strong inhibitory activity against α-amylase. On the other hand, the activity of the azuki extract component is milder than that of alkabose. Among these, C7G and E7G, whose components were isolated, showed higher activity compared to simple water extracts and heated products thereof. In particular, EG7 has higher inhibitory activity than C7G in biochemical tests.

Next, the inhibitory activity of acarbose against α-glucosidase is considerably inferior to that of all azuki extract components in biochemical tests. Looking more closely, C7G and E7G have strong inhibitory activity against α-glucosidase. Furthermore, E7G tends to have more potent inhibitory activity against α-glucosidase compared to C7G.

This is the first finding of the inhibitory effect of E7G on α-glucosidase. Therefore, the inhibitory effect on glycolytic enzymes derived from adzuki bean components, especially α-glucosidase, is useful, and when used alone or in parallel with existing drugs as an adzuki bean-derived glycolytic enzyme inhibitor, it can be used to improve blood sugar levels and metabolism. , it is expected that it will greatly contribute to the prevention and improvement of diabetes, etc.

[Formulation example]
A drug containing a glycolytic enzyme inhibitor derived from azuki bean, which is a water extract of azuki bean or its heated product, was experimentally produced. Specifically, the adzuki bean-derived glycolytic enzyme inhibitor may be a dried product (a) or a dried product (b), or a single or a mixture of C7G or E7G. The composition is disclosed in the left column, and the blending ratio (in weight percent) is disclosed in the right column. Of course, the dosage form, blending amount, etc. may be determined as appropriate.

<Formulation example 1: tablet>
Cellulose 30.00
Calcium stearate 2.50
Silicon dioxide 1.50
Starch powder 49.40
Lactose 10.00
Azuki-derived glycolytic enzyme inhibitor 6.60
(Total) 100.00

<Formulation Example 2: Capsule>
Lactose 50.00
Azuki-derived glycolytic enzyme inhibitor 50.00
(Total) 100.00
The above ingredients are encapsulated in a known capsule.

Since the azuki bean aqueous extract or the heated product of the azuki bean glycolytic enzyme inhibitor of the present invention is derived from food, it can be easily ingested through normal meals. In addition, by increasing the amount of the active ingredient and making it into a drug, it can be made into a drug that is effective in improving glucose metabolism abnormalities and is effective for prevention and treatment.
(that's all)

Claims (6)

a dipping step of soaking the azuki beans in water to obtain a dipping liquid; a dipping liquid recovery step of separating the dipping liquid and the hydrated azuki beans after the dipping step and recovering the dipping liquid; and a heating step of heating the collected dipping liquid. and contains (+)-catechin 7-O-β-D-glucopyranoside and (+)-epicatechin 7-O-β-D-glucopyranoside, which inhibits α-glucosidase enzyme activity. Method for producing enzyme inhibitors. The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to claim 1, wherein the soaking time in the soaking step is 8 to 12 hours. The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to claim 1 or 2, wherein the amount of water used for soaking the adzuki beans in the soaking step is from the same amount to twice the weight of the adzuki beans. The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to any one of claims 1 to 3, wherein the temperature in the heating step is 90 to 100°C. The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to any one of claims 1 to 4, wherein the heating time in the heating step is 60 to 120 minutes. The soaking time in the soaking step is 8 to 12 hours, the amount of water used for soaking the adzuki beans in the soaking step is the same amount to twice the weight of the beans, and the temperature in the heating step is 90°C. The method for producing an adzuki bean-derived glycolytic enzyme inhibitor according to claim 1, wherein the temperature is 100° C. and the heating time in the heating step is 60 to 120 minutes.

Images