JP2022087066A - Postprandial hyperglycemia inhibitor - Google Patents

Abstract

To provide a postprandial hyperglycemia inhibitor of solid form containing persimmon extract with high convenience and versatility for facilitating daily postprandial management of blood glucose levels.SOLUTION: A postprandial hyperglycemia inhibitor contains persimmon extract as an active ingredient.SELECTED DRAWING: None

Description

The present invention relates to an agent for suppressing an increase in postprandial blood glucose level, which comprises a persimmon extract.

Currently, the number of patients with lifestyle-related diseases is increasing due to the westernization of eating habits and chronic lack of exercise. Among them, the increase in diabetic patients is one of the most serious problems in Japan, which has entered a super-aging society. As a treatment method for diabetes, diet therapy, exercise therapy, and drug therapy are generally used, and the diet therapy is carried out with the aim of stabilizing symptoms and preventing complications by controlling blood glucose level. In order not to suffer from diabetes, it is very important to control the blood glucose level by diet therapy or the like, and in particular, the postprandial blood glucose level is attracting attention as an index related to diabetes.

When the postprandial blood glucose level during the day is not controlled, the blood glucose level during the nighttime fasting gradually deteriorates and the diabetes worsens. Therefore, it is meaningful to correct the postprandial blood glucose level. A simple blood glucose control method using daily diet instead of exercise or drugs is desired.

Polyphenols contained in natural products such as tea, grapes, apples, and cacao have various physiological activities such as antioxidant effect, blood pressure lowering effect, blood sugar level increase suppressing effect, carcinogenesis suppressing effect, and coronary heart disease improving effect. Has been reported.

The astringency of persimmons is caused by the tannins of polyphenols, and is derived from phenolic hydroxyl groups, and has antioxidant, antimutagenic (property to prevent cells from mutating), antibacterial action, and antiviral action. It is known to have many functions such as antitumor effect, antiallergic effect, blood pressure lowering effect, deodorant effect, flavor improving effect, and sickness prevention effect.

Regarding the effect of persimmon polyphenols on suppressing the increase in blood glucose level, a beverage composition containing high molecular weight persimmon tannin collected from persimmon leaf hot water extract and persimmon fruit has a molecular weight of 10, which is contained in the persimmon leaf-derived hot water extract. A beverage having an effect of suppressing an increase in postprandial blood glucose level, which contains a polyphenol extract consisting of 000 or less and persimmon tannin having a molecular weight of 10,000 or more collected from persimmon fruit, has been proposed (Patent Document 1). It was necessary to drink a certain amount of the beverage before meals, and the astringency of persimmons affected the taste of the meal, so there was room for improvement in convenience and versatility.

Further, the persimmon polyphenol oligomer described in Patent Document 2 is obtained by fragmenting persimmon tannin under acidic conditions, adsorbing it on an adsorption resin, washing it with water, eluting it with an aqueous ethanol solution, and then freeze-drying or freeze-drying the concentrated solution concentrated under reduced pressure. It is obtained as a powder by spray drying, and its main component is oligomers from dimers to pentamers. Its preparation requires complicated work and equipment, and the cost is high, so it is a versatile material. It was difficult to say.

Japanese Unexamined Patent Publication No. 2015-174829 JP-A-2007-176845

The present invention provides a solid postprandial blood glucose elevation inhibitor containing a persimmon extract, which is highly convenient and versatile for facilitating daily postprandial blood glucose control.

As a result of diligent research on the above-mentioned problems, the present inventors have found an excellent effect of suppressing an increase in blood glucose level after meals by including a persimmon extract, and completed the present invention.

Further, as a preferred embodiment of the present invention, a persimmon extract having a polyphenol content of 20% by mass or more or epigallocatechin gallate of 10% by mass or more can be used.

In the present invention, the IC50 value of the α-amylase activity inhibitory action is preferably 75 μg / mL or less. In the present invention, the single oral intake of the persimmon extract is preferably 50 mg to 5 g.

That is, the present invention provides the following gist.
[1] A solid postprandial blood glucose level inhibitor containing a persimmon extract as an active ingredient.
[2] The agent for suppressing an increase in postprandial blood glucose level according to [1], wherein the polyphenol content contained in the persimmon extract is 20% by mass or more.
[3] The agent for suppressing an increase in postprandial blood glucose level according to [1] or [2], wherein the epigallocatechin gallate contained in the persimmon extract is 10% by mass or more.
[4] The agent for suppressing an increase in postprandial blood glucose level according to any one of [1] to [3], wherein the IC50 value of the α-amylase activity inhibitory action is 75 μg / mL or less.
[5] The postprandial blood glucose level increase inhibitor according to any one of [1] to [4], which is orally ingested within 5 to 30 minutes before eating.
[6] The agent for suppressing an increase in postprandial blood glucose level according to any one of [1] to [5], wherein the single oral intake of the persimmon extract is 100 mg to 5 g.

According to the present invention, it is possible to provide an agent for suppressing an increase in postprandial blood glucose level, which comprises a persimmon extract.
The blood glucose increase inhibitor of the present invention can be ingested by humans or animals to obtain the effect of suppressing the increase in blood glucose level. That is, the persimmon extract is excellent in the activity of inhibiting enzymes such as glucositase, maltase, sucrase, and amylase. These enzymes are involved in sugar absorption from the diet, and more specifically, have an action of decomposing polysaccharides and oligosaccharides to facilitate absorption of sugar into the body. Therefore, by inhibiting the activity of these enzymes with a persimmon extract, it is possible to suppress an increase in blood glucose level, especially after a meal.

FIG. 1 is a diagram showing an HPLC chromatogram of a persimmon polyphenol thiol decomposition product. FIG. 2 is a diagram showing changes in blood glucose level after ingestion of sugar-loaded foods.

Hereinafter, the present invention will be described in detail.

(Persimmon extract and its manufacturing method)
There are no particular restrictions on the varieties of persimmon used as the raw material for persimmon extract. Examples include "Saijo", "Atago", "Ichida persimmon", and "Tenno". The site to be used is not particularly limited, and examples thereof include fruits, pericarp, flesh, seeds, leaves, rhizomes, branches, and bark. Fruits are preferable because they contain a large amount of tannin cells in which polyphenols are accumulated and can be naturally regenerated even when collected. The fruit may be used as a raw material in a state containing any other than the flesh such as shavings, pericarp and seeds as it is, or in a state where any one other than the flesh such as shavings, pericarp and seeds, or all of them are removed. It may be used as a raw material.

In order to obtain the persimmon extract, it is preferable to perform a natural and / or artificial deastringent treatment. As a result, it is insoluble in water by acetaldehyde condensation / polymerization reaction and the like. Since the formed water-insoluble matter can be easily washed with water, other isotonic liquid, a buffer solution, or the like, impurities can be effectively and efficiently removed by the washing. After washing, the polyphenol can be solubilized again by subjecting it to treatment such as heating or ultrasonic waves.

Fermentation and the like can be mentioned as a natural deastringent treatment. However, it takes one to three years or more to manufacture, and fermentation produces a strong fermented odor that causes a peculiar discomfort, so that the scene of use is likely to be limited.

As the artificial deastringent treatment, a normal method for removing astringency to sweeten the astringent persimmon can be appropriately adopted. For example, a method may be adopted in which the persimmon as a raw material is placed in a container such as a polyethylene bag together with ethanol or dry ice, sealed, and stored for a predetermined period. Alternatively, when processing a large amount of persimmon, a constant temperature short-term astringency method using carbon dioxide gas or a low-temperature astringency removal method in which astringency removal and storage are performed at the same time may be appropriately adopted. Alternatively, a simpler treatment for promoting water insolubilization of polyphenols is to crush or homogenize the raw persimmon and then store it in a container, bag, etc. that can be sealed, and 1 to 5 mL per 1 kg of the weight of the raw persimmon. It is advisable to add ethanol, seal it, and let it stand for 24 hours to 3 days. The degree of astringency can also be confirmed by a tannin printing method well known to those skilled in the art.

After the deastringent treatment, as described above, the purity of the polyphenol can be increased by washing the formed water-insoluble matter with water, an isotonic solution, a buffer solution or the like. The mode of cleaning is not particularly limited. Polyphenols, more specifically tannin cells present as tannin-accumulating cells in persimmon fruits, for example, are more separable and sedimentable in water when sufficiently deastringent. Therefore, considering that the cleaning process is efficiently performed at the lowest possible cost, the mode is that the raw material persimmon is crushed or homogenized, and the astringency is sufficiently treated with ethanol, dry ice, or the like. Then, 1 to 20 times the amount of water is added, stirred or mixed, and then allowed to stand to remove the supernatant, which is repeated 5 to 20 times, and in some cases, for a period of 5 hours to 3 days. The final precipitate can be filtered out or the like. This method makes it possible to efficiently process a relatively large amount of raw materials.

(Structure of polyphenol)
The structure of persimmon polyphenols varies depending on the variety, and epicatechin (EC), epigallocatechin gallate (ECg), epigallocatechin (EGC), and epigallocatechin gallate (EGCg) are the varieties. It is a high molecular compound having a molecular weight of about 10,000 condensed with about 30 molecules at a unique ratio, and is known to be a proanthocyanidin polymer.

(Blood sugar level rise inhibitor)
Hereinafter, the usage form of the persimmon extract obtained as described above will be described in more detail. However, the usage pattern of persimmon extract according to this explanation is not limited by the following explanation.

The postprandial blood glucose level increase inhibitor containing the persimmon extract according to the present invention is, for example, the above-mentioned persimmon extract, preferably 0.1 to 100% by mass, more preferably 1 to 80% by mass, still more preferably 30 to 70. It can be contained in% by mass.

The above-mentioned blood glucose increase inhibitor may be provided, for example, as containing persimmon extract alone, or provided as a combination of other materials or other components effective in suppressing blood glucose increase. May be.

The above-mentioned blood glucose increase inhibitor is, if necessary, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a surfactant, a lysis aid, a reducing agent, a buffer, an adsorbent, which are usually used. , Fluidizers, antistatic agents, antioxidants, sweeteners, flavoring agents, cooling agents, light-shielding agents, flavoring agents, fragrances, fragrances, coating agents, plasticizing agents and the like can be added and used.

The above-mentioned blood glucose increase inhibitor includes foods and drinks, pharmaceuticals, non-pharmaceutical products, functional foods, dietary supplements, supplements, health foods, veterinary drugs, veterinary pharmaceutical products, veterinary functional foods, and veterinary products. It can be used in the form of dietary supplements, animal supplements, animal health foods, etc., or in combination with these forms.

The method for preparing the blood glucose level increase inhibitor is not particularly limited, and the persimmon extract may be contained at least in the composition together with other components, if necessary.

The product form of the above-mentioned blood glucose increase inhibitor is not particularly limited as long as it can be easily ingested before meals. For example, powder (granule, fine granule), tablet (tablet, tablet), capsule (capsule), soft capsule. The form (soft capsule), the mode of wrapping in an edible film, and the like can be mentioned.

The persimmon extract has no problem even if it is orally ingested by humans or animals in terms of the eating experience of persimmon, and therefore, the intake of the above-mentioned blood glucose increase inhibitor is not particularly limited. The amount of intake effective for suppressing the increase in blood glucose level as described above can be appropriately determined depending on the gender, age, physique, etc. of the ingestor. For example, the amount of persimmon extract per day for an adult is 100 mg to 5 g. It is preferably present, and more preferably 50 mg to 3 g. It can be taken once to several times a day, or at any time and interval, but it may be added to foods and beverages for the purpose of suppressing sugar absorption from meals, and it is a more desirable form of use. Is preferably used so as to be taken orally 5 to 30 minutes before meals.

Hereinafter, the present invention will be described in more detail with reference to experimental examples, but these experimental examples do not limit the scope of the present invention at all.

[Preparation Example 1] (Preparation of persimmon extract 1)
The persimmon extract was prepared according to the method of Hamasaki et al. (“Rapid preparation of persimmon tannin using ethanol-deastringated fruits”, Horticultural Research (2010) 9 (3) 367-372). Specifically, it was prepared as follows. The fruits of "Tone Hayao", one of the persimmon varieties cultivated in Japan, are harvested, sealed with a 0.02 mm thick polyethylene film immediately after harvesting, and 2 mL of ethanol is added per 1 kg of fruits to 25. It was allowed to stand at ℃ for 5 days to remove astringency. The de-astringent fruits were stripped of scab and homogenized with a mixer. When the homogenized homogenate is placed in a centrifuge tube having a capacity of 50 mL and centrifuged at 1,630 × g for 15 minutes, tannin cells are layered at least below or near the bottom of the contents in the centrifuge tube. It was concentrated. The concentrated fraction was collected from a centrifuge tube and lyophilized. Next, to the freeze-dried product, 10 mL of water was added to 100 mg of the dry powder, and the mixture was heated in an autoclave (heating and pressurizing conditions: 120 ° C., 0.2 MPa) for 15 minutes. Centrifuge was performed, and the supernatant was subjected to No. 2 Filtration was obtained with a qualitative filter paper (manufactured by Advantech). The filtrate was lyophilized again to prepare a powder. When the polyphenol content of the obtained powdered persimmon extract was separately quantified by the Folin-Ciocalt method using D- (+) catechin as an index, the polyphenol content was about 70% by mass. In addition, the sugar content measured by the phenol-sulfuric acid method was about 20% by mass, and 10% by mass of other unidentified components were contained.

[Preparation Example 2] (Preparation of persimmon extract 2)
The persimmon extract was prepared as follows.

(De-astringency) Persimmons with skin and calyx were crushed and de-astringent-treated in the same manner as in Preparation Example 1. As a result, the contained polyphenols are insoluble in water by acetaldehyde condensation / polymerization reaction and the like. In addition, the contained tannin cells have improved separability and sedimentation in water.

(Washing with water) About 10 times the amount of water was added to the water-insoluble matter after the deastringent treatment for washing. Washing was repeated 10 times in total over 2 days. As a result, components other than polyphenols can be often removed.

(Water / heating) Approximately 10 times the amount of water was added to the water-insoluble product after washing with water, and the mixture was treated in an autoclave (heating and pressurizing conditions: 120 ° C., 0.2 MPa) for 1 hour. As a result, the contained polyphenol breaks the bonds such as aldehyde condensation and polymerization, and the polyphenol is solubilized in water.

(Squeezing / filtering) The heated processed product was rubbed with a cloth, passed through a 50 μm filter, and finally passed through a 1 μm filter. The Brix value of the obtained filter-passing liquid was about Brix1.

(Concentration / pulverization) The liquid passing through the filter was concentrated about 15 times with a vacuum concentrator. This was drum-dried, pulverized and sieved to obtain a 250 μm mesh pass powder. The polyphenol concentration of this powder was quantified by the Folin-Ciocalt method using D- (+) catechin as an index, and was 70% by weight in terms of catechin.

[Measurement of catechins constituting polyphenols]
<Test sample>
Experimental Example 1: Persimmon extract ("Nara-style persimmon astringent (powder)" manufactured by Ishii Bussan Co., Ltd.)
Experimental Example 2: Apple polyphenol ("Apple Phenon SH" manufactured by Asahi Group Foods Co., Ltd.)
Experimental Example 3: Tea polyphenol ("Sanphenon 90S" manufactured by Taiyo Kagaku Co., Ltd.)
Experimental Example 4: Lychee Polyphenol ("OLG-F" manufactured by Amino Up Co., Ltd.)

<Test method>
The polyphenols contained in each test sample were dissolved in ethanol (containing 1% 2-mercaptoethanol and 0.5% HCl) to 25 mg / mL and reacted at 40 ° C. for 4 days. Then, HPLC analysis was performed under the following conditions. The analysis results are shown in FIG.

<Analysis conditions>
Column: ODS 4.6 x 150 mm, 5 μm, column temperature 35 ° C (manufactured by GL Sciences Co., Ltd.)
Flow velocity: 1.0 mL / min Wavelength: 280 nm
Mobile phase: Liquid A (2% aqueous acetic acid solution), liquid B (methanol)
min 0-5 5-55 55-60
A: B 10: 0 2: 8 10: 0

The proportion of constituent catechins was calculated by the following method. The thiol decomposition products of catechins (EGC, EC, EGCg, ECg) were identified by the LC-MS method, and the area value calculated by the HPLC method and the molar extinction coefficient (Kinoshita et al., Thrombosis hemostatic magazine 4 (6) 417-422 , 1993), the molar ratio was calculated, and the ratio to the total value of catechins was calculated. The results are shown in Table 2. The molar extinction coefficient of catechins at 280 nm was EGC; 1,100, EC; 3,300, EGCg; 8,700, ECg; 13,000.

構成カテキンの割合（モル比）

Percentage of constituent catechins (molar ratio)

As shown in Table 2, it can be seen that the persimmon extract has the highest catechin composition ratio of epigallocatechin gallate (EGCg) as compared with other polyphenol materials.

[Measurement of α-amylase activity inhibitory effect]
<Test sample>
Experimental Example 5: Persimmon extract ("Nara-style persimmon astringent (powder)" manufactured by Ishii Bussan Co., Ltd.) (Test solution concentration: 0.13, 0.25, 0.42 mg / mL 50% ethanol aqueous solution)
Experimental Example 6: Saracia ("Saracia oblonga extract powder B" manufactured by Takama Co., Ltd.) (Test solution concentration: 0.5, 1.5, 5.0 mg / mL 25% DMSO aqueous solution)
Experimental Example 7: Mulberry leaf extract ("DNJ Extract Powder RF" manufactured by Toyoda Kaori Co., Ltd.) (Test solution concentration: 1.0, 3.0, 10 mg / mL water)
Experimental Example 8: Acarbose (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd., for pharmacological research) (test solution concentration: 0.06, 0.12, 0.18 mg / mL water), positive control

<Test method>
An α-amylase measurement kit (manufactured by Kikkoman Biochemifa Co., Ltd.) and α-Amylase from Porcine pancreas (manufactured by Sigma-Aldrich) were used to measure the α-amylase inhibitory effect. Equal amounts of the substrate solution and the enzyme solution (included in the kit) were mixed (substrate-enzyme mixed solution), and the mixture was heated at 37 ° C. for 5 minutes. An equal amount mixture of α-amylase and the test solution of each example was added in an amount of 1/10 of the substrate-enzyme mixed solution and reacted for 10 minutes. Then, the reaction was stopped by adding twice the amount of the substrate-enzyme mixed solution (included in the kit) to stop the reaction, and the absorbance at 400 nm was measured. For the blank, a mixed solution of α-amylase solution and test solution was added after the reaction was stopped, and the absorbance at 400 nm was measured. The α-amylase inhibition rate was calculated by the following formula, and the IC50 value of the α-amylase activity inhibitory action was determined.
α-amylase inhibition rate (%) = {1- (A-Blank A) / (B-Blank B)} × 100
A: Absorbance of the sample addition reaction system
Blank A: Absorbance of the blank in the sample addition reaction system
B: Absorbance of the sample solvent addition reaction system (control: total color development)
Blank B: Absorbance of the blank of the sample solvent addition reaction system

<Test results>
The results are shown in Table 1.

α-アミラーゼ活性阻害作用のＩＣ５０値

IC50 value of α-amylase activity inhibitory action

The IC50 value (μg / mL) of the α-amylase activity inhibitory action is evaluated to be higher when the value is lower. It can be seen that the persimmon extract is inferior to acarbose, which is a prescription drug for treating type 2 diabetes, but has an excellent effect compared to salasia and mulberry leaf extract, which are said to have an effect of suppressing an increase in blood glucose level. ..

[Measurement of postprandial blood glucose elevation inhibitory effect]
<Test method>
A crossover study was conducted on 11 healthy adult male and female subjects to measure the effect of persimmon extract on postprandial blood glucose elevation. For the persimmon extract ingestion group, 100 mg of persimmon extract powder wrapped in an edible film (manufactured by Pip Co., Ltd., bag oblate) was ingested as a test meal, and for the persimmon extract non-intake group, only the above edible film was ingested for 20 minutes. Later, a sugar-loaded food (about 100 g of a commercially available salted rice ball) was ingested. The blood glucose level was measured using a glucose meter (Terumo Corporation, Medisafe Fit MS-FR201B) before ingestion of the test meal or edible film alone, and 30, 60, 90, 120 minutes after ingestion of the sugar-loaded food. .. The test was conducted in the morning, and eating, drinking and smoking other than water was prohibited after 21:00 the day before, and it was continued until the end of the test. FIG. 2 shows the average value of changes in blood glucose level from before ingestion to 120 minutes after ingestion, based on the blood glucose level before ingestion of the test meal or the edible film alone.

<Test results>
As shown in the test results of FIG. 2, the change in blood glucose level 60 minutes and 90 minutes after ingesting the sugar-loaded food was significantly lower in the persimmon extract ingested group than in the persimmon extract non-ingested group. The effect of suppressing the increase in postprandial blood glucose level was observed.

Claims (6)

A solid postprandial blood glucose increase inhibitor characterized by containing persimmon extract as an active ingredient. The agent for suppressing an increase in postprandial blood glucose level according to claim 1, wherein the polyphenol content contained in the persimmon extract is 20% by mass or more. The agent for suppressing an increase in postprandial blood glucose level according to claim 1 or 2, wherein the epigallocatechin gallate contained in the persimmon extract is 10% by mass or more. The agent for suppressing an increase in postprandial blood glucose level according to any one of claims 1 to 3, wherein the IC50 value of the α-amylase activity inhibitory action is 75 μg / mL or less. The postprandial blood glucose level increase inhibitor according to any one of claims 1 to 4, which is orally ingested within 5 to 30 minutes before eating. The agent for suppressing an increase in postprandial blood glucose level according to any one of claims 1 to 5, wherein the single oral intake of the persimmon extract is 100 mg to 5 g.

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