CN113546074A

CN113546074A - Composition containing 3-O-methyl quercetin and application thereof in alpha-glucosidase inhibition

Info

Publication number: CN113546074A
Application number: CN202110961983.5A
Authority: CN
Inventors: 张强; 焦中高; 陈大磊; 刘杰超; 刘慧�; 张春岭; 杨文博; 吕真真; 潘俊坤
Original assignee: Zhengzhou Fruit Research Institute CAAS
Current assignee: Zhengzhou Fruit Research Institute CAAS
Priority date: 2021-08-20
Filing date: 2021-08-20
Publication date: 2021-10-26
Anticipated expiration: 2041-08-20
Also published as: CN113546074B

Abstract

The invention discloses a composition containing 3-O-methyl quercetin and application thereof in alpha-glucosidase inhibition, belonging to the technical field of natural active compounds. The composition of the invention comprises 3-O-methyl quercetin and one of dihydroquercetin and calycosin; wherein the mass ratio of the 3-O-methyl quercetin to the dihydroquercetin is 2:25-4: 25; the mass ratio of the 3-O-methyl quercetin to the calycosin is 2: 6. The composition has obvious synergistic effect of inhibiting alpha-glucosidase, has better effect than that of singly using the flavone compound, can reduce the dosage of the medicament, and reduces the occurrence of medicament resistance.

Description

Composition containing 3-O-methyl quercetin and application thereof in alpha-glucosidase inhibition

Technical Field

The invention belongs to the technical field of natural active compounds, and particularly relates to a composition containing 3-O-methyl quercetin and application thereof in alpha-glucosidase inhibition.

Background

In recent years, much research has been devoted to identify effective α -glucosidase inhibitors from natural sources and to develop antidiabetic drugs or lead compounds, including flavonoids, anthocyanins, phenols, and the like. Alpha-glucosidase inhibitors can delay the release of D-glucose from food carbohydrates and delay glucose absorption, resulting in a decrease in postprandial plasma glucose levels, thereby effectively controlling postprandial hyperglycemia.

The natural flavonoid exists in various fruits and vegetables such as grapes and blueberries, and researches report that the natural flavonoid has various biological activities such as blood sugar reduction, cancer resistance and oxidation resistance. Currently, the clinical drugs widely used for treating type II diabetes mellitus are oral hypoglycemic drugs such as acarbose, voglibose and miglitol. The medicine can effectively improve and prevent hyperglycemia and complications thereof by acting with alpha-glucosidase in small intestine, inhibiting enzyme activity, delaying glucose production, preventing postprandial blood sugar from reaching a certain peak value. However, these inhibitors cause a series of side effects, such as diarrhea, abdominal pain, liver injury, etc., some patients are not fully accepted, and new synthetic drugs have certain toxic and side effects and low efficiency, so that the search for alpha-glucosidase inhibitors derived from natural plants becomes one of the main directions for the current research and treatment of type II diabetes drugs (Guoshaxia, Zengyang, Xueyeng, etc.. the pharmacological research progress of alpha-glucosidase inhibitors [ J ]. university of Qinghai university: Nature science edition, 2011,27(1): 63-66).

The strategy of drug combination is very important, on one hand, the sensitivity of the drug can be increased, and the drug resistance problem can be solved; on the other hand, the dosage can be reduced, and the toxic and side effects of the medicine can be reduced. At present, most researches on alpha-glucosidase inhibitors focus on a single compound, certain side effects and tolerance can be generated by continuous use, and the reports of synergistic effect among active molecules are few. Therefore, the research on the combined application of the flavonoid compounds to inhibit alpha-glucosidase and improve the hypoglycemic activity has important significance for improving the human health.

Disclosure of Invention

The invention aims to provide a composition containing 3-O-methyl quercetin and application thereof in alpha-glucosidase inhibition, so as to solve the problems that in the prior art, a single active ingredient has limited effect of reducing blood sugar and is easy to generate drug resistance.

In order to achieve the purpose, the invention is realized by the following technical scheme:

a composition containing 3-O-methyl quercetin comprises 3-O-methyl quercetin and one of dihydroquercetin and calycosin;

wherein the mass ratio of the 3-O-methyl quercetin to the dihydroquercetin is 2:25-4: 25; the mass ratio of the 3-O-methyl quercetin to the calycosin is 2: 6.

In several embodiments, the mass ratio of 3-O-methyl quercetin to dihydroquercetin is 2:25, 4: 25; wherein when the mass ratio of 3-O-methyl quercetin to dihydroquercetin is 2:25, the combined medication index mean value (CI) is_avg) Is 0.50, is better than the mass ratio of 4:25, and has stronger synergistic effect.

The application of the composition in preparing preparations with alpha-glucosidase inhibiting effect.

An alpha-glucosidase inhibitor contains 3-O-methyl quercetin and one of dihydroquercetin and calycosin as effective components; wherein the mass ratio of the 3-O-methyl quercetin to the dihydroquercetin is 2:25-4: 25; the mass ratio of the 3-O-methyl quercetin to the calycosin is 2: 6.

The application of the composition in preparing a medicament with the hypoglycemic effect is to achieve the purpose of controlling postprandial hyperglycemia by inhibiting the activity of alpha-glucosidase and blocking the digestion and absorption of carbohydrate.

A medicine with blood sugar lowering effect contains 3-O-methyl quercetin and dihydroquercetin or 3-O-methyl quercetin and calycosin as effective components; wherein the mass ratio of the 3-O-methyl quercetin to the dihydroquercetin is 2:25-4: 25; the mass ratio of the 3-O-methyl quercetin to the calycosin is 2: 6.

Within the limited mass ratio range, the 3-O-methyl quercetin and dihydroquercetin, the 3-O-methyl quercetin and the calycosin achieve the synergistic technical effect.

The medicine comprises pharmaceutically acceptable carriers, solvents, diluents, excipients and other media which are mixed, and can be prepared into powder, granules, capsules, injections, oral liquid or tablets according to different requirements.

The technical scheme of the invention has the advantages

The composition of the 3-O-methyl quercetin, the dihydroquercetin, the 3-O-methyl quercetin and the calycosin has obvious synergistic effect of inhibiting alpha-glucosidase, has better effect than that of singly using the flavone compound, can reduce the dosage of the medicine and reduce the occurrence of drug resistance. When dihydroquercetin and calycosin are replaced by quercetin and formononetin with similar chemical structures, the synergistic effect disappears.

Through an in vitro alpha-glucosidase inhibition test and by applying a Chou-Talalay method, the composition of the 3-O-methyl quercetin, the dihydroquercetin, the 3-O-methyl quercetin and the calycosin of the invention is proved to have obvious synergistic effect on the alpha-glucosidase when the mass ratio is 2:25-4:25 and 2:6 respectively, and the content is 50% (GI)₅₀)、75％(GI₇₅) And 90% (GI)₉₀) The CI value at the inhibition rate is less than 1.0, and the medicine at the high inhibition rateThe intensity of the synergistic effect is generally higher than that of the inhibition rate.

Drawings

FIG. 1 the inhibitory activity of the 3-O-methyl quercetin compositions of example 1 and example 4 on α -glucosidase;

FIG. 23 is a graph of the inhibition of alpha-glucosidase by the O-methyl Quercetin + DihydroQuercetin (2:25) composition;

FIG. 33 is a graph of Fa-CI trends for alpha-glucosidase inhibition by the-O-methyl quercetin + dihydroquercetin (2:25) composition;

FIG. 43 is a graph of the inhibition of alpha-glucosidase by the O-methyl quercetin + calycosin (2:6) composition;

FIG. 53 is a graph of the Fa-CI trend for the O-methyl quercetin + calycosin (2:6) composition to inhibit alpha-glucosidase;

FIG. 6 the inhibitory activity of the 3-O-methyl quercetin compositions of example 2 and example 5 on α -glucosidase;

FIG. 73 is a graph of the inhibition of alpha-glucosidase by the-O-methyl quercetin + dihydroquercetin (4:25) composition;

FIG. 83 is a graph of the inhibition of alpha-glucosidase by the O-methyl quercetin + calycosin (4:6) composition;

FIG. 9 the inhibitory activity of the 3-O-methyl quercetin compositions of example 3 and example 6 on α -glucosidase;

FIG. 103 is a graph of the inhibition of alpha-glucosidase by the O-methyl quercetin + dihydroquercetin (2:30) composition;

FIG. 113-graph of the inhibition of alpha-glucosidase by O-methyl Quercetin + Calycosin (2:10) compositions;

FIG. 12 the inhibitory activity of the 3-O-methyl quercetin compositions of comparative example 1 and comparative example 2 on α -glucosidase;

FIG. 133-O-methyl Quercetin + Quercetin (2:40) composition inhibition of α -glucosidase plot;

FIG. 143-O-methyl Quercetin + formononetin (2:20) composition is a graph of the inhibition of alpha-glucosidase.

Detailed Description

Terms used in the present invention have generally meanings as commonly understood by one of ordinary skill in the art, unless otherwise specified.

3-O-Methyl Quercetin (3-O-Methyl Quercetin) is one of Quercetin derivatives, and has molecular formula of C₁₆H₁₂O₇(ii) a Molecular weight: 316.26, respectively; CAS accession number: 1486-70-0, structural formula:

dihydroquercetin (Taxifolin) with molecular formula of C₁₅H₁₂O₇(ii) a Molecular weight: 304.25, respectively; CAS accession number: 480-18-2, the structural formula is as follows:

calycosin (Calycosin) with molecular formula of C₁₆H₁₂O₅(ii) a Molecular weight: 284.26, respectively; CAS accession number: 20575-57-9, structural formula:

alpha-glucosidase (from saccharomyces cerevisiae, Sigma);

4-nitrobenzene- α -D-glucopyranoside (pNPG, TOKYO chemical Industry co., LTD);

acarbose (TOKYO chemical Industry co., LTD);

3-O-methyl quercetin, dihydroquercetin, calycosin (beijing solibao);

millipore silicon water purification system (Millipore, france);

sodium phosphate salt buffer (pH 6.8,0.1mol L)^-1)；

The microplate reader TECAN infinite M200 PRO (Teacan Group ltd., Swizerland).

The present invention will be described in further detail with reference to the following data in conjunction with specific examples. The following examples are intended to illustrate the invention and are not intended to limit the scope of the invention in any way.

Example 1

A composition of 3-O-methyl quercetin and dihydroquercetin, wherein the mass ratio of the 3-O-methyl quercetin to the dihydroquercetin is 2: 25; specifically, the concentrations of 3-O-methyl quercetin and dihydroquercetin in the composition are 2 μ g/mL and 25 μ g/mL, respectively.

Example 2

A composition of 3-O-methyl quercetin and dihydroquercetin, wherein the mass ratio of the 3-O-methyl quercetin to the dihydroquercetin is 4: 25; specifically, the concentrations of 3-O-methyl quercetin and dihydroquercetin in the composition are 4 μ g/mL and 25 μ g/mL, respectively.

Example 3

A composition of 3-O-methyl quercetin and dihydroquercetin, wherein the mass ratio of the 3-O-methyl quercetin to the dihydroquercetin is 2: 30; specifically, the concentrations of 3-O-methyl quercetin and dihydroquercetin in the composition are 2 μ g/mL and 30 μ g/mL, respectively.

Example 4

A composition of 3-O-methyl quercetin and calycosin, wherein the mass ratio of the 3-O-methyl quercetin to the calycosin is 2: 6; specifically, the concentrations of 3-O-methyl quercetin and calycosin in the composition are 2 μ g/mL and 6 μ g/mL respectively.

Example 5

A composition of 3-O-methyl quercetin and calycosin, wherein the mass ratio of the 3-O-methyl quercetin to the calycosin is 4: 6; specifically, the concentrations of 3-O-methyl quercetin and calycosin in the composition are 4 μ g/mL and 6 μ g/mL respectively.

Example 6

A composition of 3-O-methyl quercetin and calycosin, wherein the mass ratio of the 3-O-methyl quercetin to the calycosin is 2: 10; specifically, the concentrations of 3-O-methyl quercetin and calycosin in the composition are 2 μ g/mL and 10 μ g/mL respectively.

Hypoglycemic Effect test of 3-O-methyl Quercetin composition

The experimental method comprises the following steps:

with 0.1mol L^-1PBS buffer solution with pH 6.8 is used for preparing alpha-glucosidase solution with concentration of 0.25U/mL and substrate p-nitrobenzene-alpha-D-glucopyranoside (pNPG) solution with concentration of 5 mmol/mL.

Add 100. mu.L of sample solution per well followed by 40. mu.L of alpha-glucosidase (0.25U/mL), react at 37 ℃ for 15min, then add 60. mu.L of the 5mmol/mL substrate p-nitrophenyl-alpha-D-glucopyranoside (pNPG); after leaving at 37 ℃ for reaction for 15min, the reaction was measured at a wavelength of 405nm with a microplate reader.

The sample solution to be tested is the 3-O-methyl quercetin composition described in the embodiment 1-6, firstly, dimethyl sulfoxide (DMSO) is adopted to prepare 3-O-methyl quercetin, dihydroquercetin and calycosin into mother liquor of 10mg/mL respectively; and then PBS buffer solution is used for preparing 3-O-methyl quercetin, dihydroquercetin, calycosin and the sample solution of the composition with specific concentration.

The positive control was acarbose (350. mu.g/mL), the blank was without sample and enzyme, and the sample blank was without enzyme.

Calculating the formula: inhibition rate [1- (OD)_{Sample (I)}–OD_{Sample blank})/(OD_{Negative control}-OD_{Blank space})]×100％

The CI values were calculated according to the software CompuSyn to evaluate the synergy between drugs.

Combination Index (CI) was used to describe the magnitude of drug synergy: CI <1 represents that the medicines have synergistic effect, the curative effect of each monomer medicine can be enhanced by combined medication, and the synergistic effect is stronger when the CI value is smaller; CI-1 represents that the drugs have additive effect, and the combined drug result is only the linear superposition of the curative effect of each monomer drug; CI >1 represents antagonism among the drugs, and the combination of the drugs can reduce the respective curative effect.

1. The 3-O-methyl quercetin compositions of example 1 and example 4 had alpha-glucosidase inhibitory activity

The α -glucosidase inhibitory activity of the 3-O-methyl quercetin compositions of example 1 and example 4 is shown in FIG. 1: the inhibition rates of 2 mu g/mL of 3-O-methyl quercetin, 25 mu g/mL of dihydroquercetin, 6 mu g/mL of calycosin and 350 mu g/mL of acarbose on alpha-glucosidase under corresponding mass concentrations are 46.23 +/-1.2%, 40.12 +/-2.1%, 56.6 +/-4.3% and 46.25 +/-3.5% respectively; the inhibition rate of the 3-O-methyl quercetin and dihydroquercetin composition (2+25 mug/mL) is 78.82 +/-3.4%, and the inhibition rate of the 3-O-methyl quercetin and the calycosin (2+6 mug/mL) is 73.21 +/-4.5%; the results show that the composition remarkably improves the inhibitory activity to alpha-glucosidase when the composition is used in combination.

Detecting the alpha-glucosidase inhibitory activity of a 3-O-methyl quercetin and dihydroquercetin composition with a mass ratio of 2:25 under different concentration gradients, wherein the concentration gradient of the 3-O-methyl quercetin and dihydroquercetin composition is (mu g/mL): 2+25, 1+12.5, 0.5+6.25, 0.25+ 3.125; the concentration gradient of 3-O-methyl quercetin was (μ g/mL): 2.1, 0.5, 0.25; the concentration gradient of dihydroquercetin is (mu g/mL): 25. 12.5, 6.25, 3.125; the results are shown in FIG. 2; the 3-O-methyl quercetin and dihydroquercetin composition with the mass ratio of 2:25 improves the inhibition activity on alpha-glucosidase under different concentration gradients. The Fa-CI trend of the 3-O-methyl quercetin and dihydroquercetin composition at a mass ratio of 2:25 is shown in FIG. 3, and it can be seen from FIG. 3 that the CI values of both 3-O-methyl quercetin and dihydroquercetin are below 1.0, showing a synergistic effect.

Detecting the alpha-glucosidase inhibitory activity of the 3-O-methyl quercetin and calycosin composition with the mass ratio of 2:6 under different concentration gradients, wherein the concentration gradient of the 3-O-methyl quercetin and calycosin composition is (mu g/mL): 2+6, 1+3, 0.5+1.5, 0.25+ 0.75; the concentration gradient of 3-O-methyl quercetin was (μ g/mL): 2.1, 0.5, 0.25; calycosin concentration gradient (μ g/mL): 6. 3, 1.5, 0.75; the results are shown in FIG. 4: the 3-O-methyl quercetin and calycosin composition with the mass ratio of 2:6 also correspondingly improves the inhibitory activity to alpha-glucosidase under different concentration gradients. The Fa-CI trend of the 3-O-methylquercetin and calycosin composition at a mass ratio of 2:6 is shown in FIG. 5, and it is understood from FIG. 5 that the CI values of both 3-O-methylquercetin and calycosin are 1.0 or less, showing a synergistic effect.

The combination Coefficient (CI) of the 3-O-methyl quercetin compositions of example 1 and example 4 is shown in Table 1:

TABLE 1 Combined administration Coefficients (CI) of the 3-O-methyl quercetin compositions of example 1 and example 4

Data are derived from the results of three independent experiments, expressed as mean ± standard deviation

As is clear from the results in Table 1, the combined administration coefficients CI of 3-O-methyl quercetin and dihydroquercetin (2:25) and 3-O-methyl quercetin and calycosin (2:6) when used in combination are all less than 1, showing a synergistic effect, wherein the combined administration coefficient of the 3-O-methyl quercetin and dihydroquercetin composition in GI₅₀，GI₇₅And GI₉₀Are all less than 0.60, show strong synergistic effect, and have combined medication index mean value (CI)_avg) Is 0.50; the combined administration coefficient of 3-O-methyl quercetin and verbascoisoflavone composition in GI₇₅And GI₉₀Are all less than 0.9, show synergistic effect, and have mean value of combined medication index (CI)_avg) Is 0.80.

2. The 3-O-methyl quercetin compositions of example 2 and example 5 had alpha-glucosidase inhibitory activity

The α -glucosidase inhibitory activity of the 3-O-methyl quercetin compositions of example 2 and example 5 is shown in FIG. 6: the inhibition rates of 4 mu g/mL of 3-O-methyl quercetin, 25 mu g/mL of dihydroquercetin, 6 mu g/mL of calycosin and 350 mu g/mL of acarbose on alpha-glucosidase under corresponding mass concentrations are 53.52 +/-3.2%, 40.12 +/-2.1%, 56.6 +/-4.3% and 46.25 +/-3.5% respectively; the inhibition rate of the 3-O-methyl quercetin and dihydroquercetin composition (4+25 mug/mL) is 61.82 +/-4.4%, and the inhibition rate of the 3-O-methyl quercetin and calycosin (4+6 mug/mL) is 55.54 +/-3.5%; the results show that the composition improves the inhibition activity to alpha-glucosidase when the 3-O-methyl quercetin and the dihydroquercetin are used together; the composition does not significantly improve the alpha-glucosidase inhibitory activity when the 3-O-methyl quercetin and calycosin are used in combination.

Detecting the alpha-glucosidase inhibitory activity of a 3-O-methyl quercetin and dihydroquercetin composition with a mass ratio of 4:25 under different concentration gradients, wherein the concentration gradient of the 3-O-methyl quercetin and dihydroquercetin composition is (mu g/mL): 4+25, 2+12.5, 1+6.25, 0.5+ 3.125; the concentration gradient of 3-O-methyl quercetin was (μ g/mL): 4.2, 1, 0.5; the concentration gradient of dihydroquercetin is (mu g/mL): 25. 12.5, 6.25, 3.125; the results are shown in FIG. 7.

Detecting the alpha-glucosidase inhibitory activity of the 3-O-methyl quercetin and calycosin composition with the mass ratio of 4:6 under different concentration gradients, wherein the concentration gradient of the 3-O-methyl quercetin and calycosin composition is (mu g/mL): 4+6, 2+3, 1+1.5, 0.5+ 0.75; the concentration gradient of 3-O-methyl quercetin was (μ g/mL): 4.2, 1, 0.5; calycosin concentration gradient (μ g/mL): 6. 3, 1.5, 0.75; the results are shown in FIG. 8.

The combination Coefficient (CI) of the 3-O-methyl quercetin compositions of example 2 and example 5 is shown in Table 2:

TABLE 2 Combined administration Coefficients (CI) of the 3-O-methyl quercetin compositions of example 2 and example 5

As can be seen from the results in Table 2, the co-administration coefficients CI of 3-O-methyl quercetin and dihydroquercetin (4:25) were all less than 1, and showed synergistic effects, and the mean co-administration index (CI)_avg) Is 0.75; the 3-O-methyl quercetin and calycosin (4:6) have combined administration coefficients CI of more than 1, and show antagonism.

3. The 3-O-methyl quercetin compositions of example 3 and example 6 have alpha-glucosidase inhibitory activity

The α -glucosidase inhibitory activity of the 3-O-methyl quercetin compositions of example 3 and example 6 is shown in FIG. 9: the inhibition rates of 2 mu g/mL of 3-O-methyl quercetin, 30 mu g/mL of dihydroquercetin, 10 mu g/mL of calycosin and 350 mu g/mL of acarbose on alpha-glucosidase under corresponding mass concentrations are 46.23 +/-1.2%, 54.5 +/-4.1%, 62.12 +/-3.3% and 46.25 +/-3.5% respectively; the inhibition rate of the 3-O-methyl quercetin and dihydroquercetin composition (2+30 mug/mL) is 57.62 +/-5.7%, and the inhibition rate of the 3-O-methyl quercetin and the stamen isoflavone (2+10 mug/mL) is 56.21 +/-4.5%; the results show that the combination does not obviously improve the inhibition activity to alpha-glucosidase when used together.

Detecting the alpha-glucosidase inhibitory activity of a 3-O-methyl quercetin and dihydroquercetin composition with a mass ratio of 2:30 under different concentration gradients, wherein the concentration gradient of the 3-O-methyl quercetin and dihydroquercetin composition is (mu g/mL): 2+30, 1+15, 0.5+7.5, 0.25+ 3.75; the concentration gradient of 3-O-methyl quercetin was (μ g/mL): 2.1, 0.5, 0.25; the concentration gradient of dihydroquercetin is (mu g/mL): 30. 15, 7.5, 3.75; the results are shown in FIG. 10.

Detecting the alpha-glucosidase inhibitory activity of the 3-O-methyl quercetin and calycosin composition with the mass ratio of 2:10 under different concentration gradients, wherein the concentration gradient of the 3-O-methyl quercetin and calycosin composition is (mu g/mL): 2+10, 1+5, 0.5+2.5, 0.25+ 1.25; the concentration gradient of 3-O-methyl quercetin was (μ g/mL): 2.1, 0.5, 0.25; calycosin concentration gradient (μ g/mL): 10. 5, 2.5, 1.25; the results are shown in FIG. 11.

The combination Coefficient (CI) of the 3-O-methylquercetin compositions of example 3 and example 6 is shown in Table 3:

TABLE 3 Combined administration Coefficients (CI) of the 3-O-methyl quercetin compositions of example 3 and example 6

As is clear from the results in Table 3, the combination of 3-O-methylquercetin and dihydroquercetin (2:30) and 3-O-methylquercetin and calycosin (2:10) showed an antagonistic effect with a combination coefficient CI of greater than 1.

Comparative example 1

Quercetin (Quercitrin) has a structure similar to that of dihydroquercetin, and has a molecular formula of C₂₁H₂₀O₁₁(ii) a Molecular weight: 448.38, respectively; CAS accession number: 522-12-3, the structural formula is:

the composition comprises 3-O-methyl quercetin and quercetin, wherein the mass ratio of 3-O-methyl quercetin to quercetin is 2:40, and specifically, the concentrations of 3-O-methyl quercetin and quercetin in the composition are 2 μ g/mL and 40 μ g/mL respectively.

Comparative example 2

Formononetin (Formononetin) with a molecular formula of C and a structure similar to calycosin₁₆H₁₂O₄(ii) a Molecular weight: 268.26, respectively; CAS accession number: 485-72-3, the structural formula is:

the composition of 3-O-methyl quercetin and formononetin, wherein the mass ratio of the 3-O-methyl quercetin to the formononetin is 2:20, and specifically, the concentrations of the 3-O-methyl quercetin and the formononetin in the composition are respectively 2 mu g/mL and 20 mu g/mL.

Hypoglycemic Effect test of the compositions of comparative examples 1 and 2

The detection of the α -glucosidase inhibitory activity of the 3-O-methyl quercetin compositions of comparative example 1 and comparative example 2 is shown in FIG. 12: the inhibition rates of 2 mu g/mL of 3-O-methyl quercetin, 40 mu g/mL of quercetin, 20 mu g/mL of formononetin and 350 mu g/mL of acarbose on alpha-glucosidase under corresponding mass concentrations are 46.23 +/-1.2%, 53.52 +/-3.2%, 60.07 +/-5.3% and 46.25 +/-3.5% respectively; the inhibition rate of the 3-O-methyl quercetin and quercetin composition (2+40 mu g/mL) is 60.8 +/-3.4%; the inhibition rate of the 3-O-methyl quercetin and formononetin composition (2+20 mu g/mL) is 63.3 +/-5.1%; the results show that the two compositions do not obviously improve the inhibition activity to alpha-glucosidase when used together.

Detecting the alpha-glucosidase inhibitory activity of a 3-O-methyl quercetin and quercitrin composition with the mass ratio of 2:40 under different concentration gradients, wherein the concentration gradient of the 3-O-methyl quercetin and quercitrin composition is (mu g/mL): 2+40, 1+20, 0.5+10, 0.25+ 5; the concentration gradient of 3-O-methyl quercetin was (μ g/mL): 2.1, 0.5, 0.25; the concentration gradient of quercetin was (μ g/mL): 40. 20, 10, 5; the results are shown in FIG. 13.

Detecting the alpha-glucosidase inhibitory activity of a 3-O-methyl quercetin and formononetin composition with a mass ratio of 2:20 under different concentration gradients, wherein the concentration gradient of the 3-O-methyl quercetin and formononetin composition is (mu g/mL): 2+20, 1+10, 0.5+5, 0.25+ 2.5; the concentration gradient of 3-O-methyl quercetin was (μ g/mL): 2.1, 0.5, 0.25; the concentration gradient of formononetin is (mu g/mL): 20. 10, 5, 2.5; the results are shown in FIG. 14.

TABLE 4 Combined dosing factor (CI) for the 3-O-methyl quercetin compositions of comparative example 1 and comparative example 2

As is clear from the results in Table 4, the combination coefficient CI of 3-O-methylquercetin and quercetin (2:40), and 3-O-methylquercetin and formononetin (2:20) were greater than 1, and the antagonism was exhibited.

The foregoing is directed to preferred embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.

Claims

1. A composition containing 3-O-methyl quercetin, characterized by comprising 3-O-methyl quercetin and one of dihydroquercetin, calycosin;

2. Use of the composition of claim 1 for the preparation of a preparation having an alpha-glucosidase inhibitory effect.

3. An alpha-glucosidase inhibitor, characterized in that the effective component comprises 3-O-methyl quercetin and one of dihydroquercetin and calycosin.

4. The α -glucosidase inhibitor according to claim 3, wherein the mass ratio of 3-O-methyl quercetin to dihydroquercetin is 2:25-4: 25; the mass ratio of the 3-O-methyl quercetin to the calycosin is 2: 6.

5. Use of a composition according to claim 1 for the preparation of a medicament having a hypoglycemic effect.

6. The use according to claim 5, wherein the hypoglycemic effect is to control postprandial hyperglycemia by inhibiting the activity of α -glucosidase and blocking the digestion and absorption of carbohydrates.

7. A medicine with blood sugar lowering effect is characterized in that the effective component comprises 3-O-methyl quercetin and one of dihydroquercetin and calycosin; wherein the mass ratio of the 3-O-methyl quercetin to the dihydroquercetin is 2:25-4: 25; the mass ratio of the 3-O-methyl quercetin to the calycosin is 2: 6.