A hypoglycemic compound and its application

Abstract

The present invention provides a hypoglycemic compound and its application. The hypoglycemic compound includes quercetin and 3-O-methylquercetin. The present invention achieves a synergistic effect through reasonable compounding, achieving blood sugar lowering, reducing dosage and side effects while simultaneously lowering blood sugar, and avoiding antagonistic reactions between the two components.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a blood sugar lowering compound and application thereof. Background Art

[0002] Currently, diabetes, especially type 2 diabetes and its complications, pose an increasingly serious threat to human health. α-glucosidase hydrolyzes oligosaccharides to release glucose, ultimately leading to postprandial hyperglycemia. Therefore, inhibiting α-glucosidase activity, thereby delaying glucose production at the source and reducing postprandial hyperglycemia, is a major approach to treating diabetes. Since currently used clinical α-glucosidase inhibitors have certain side effects with long-term use, it is particularly important to identify safe, naturally derived inhibitors that can inhibit α-glucosidase with minimal toxicity and side effects.

[0003] Flavonoids are commonly found in various natural plants and Chinese herbal medicines. They have minimal toxicity and possess diverse pharmacological activities. Screening for effective α-glucosidase inhibitors from natural plant active ingredients such as flavonoids has become a research hotspot. However, it is common for two or more drug ingredients in food or health supplements to produce similar effects. Therefore, the goal of drug therapy is to improve efficacy while minimizing side effects when different drugs are used in combination. Summary of the Invention

[0004] The purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide a hypoglycemic compound that achieves a synergistic effect through reasonable compounding, thereby achieving the goal of lowering blood sugar while reducing the dosage and side effects, and avoiding the mutual reaction between the two components to produce an antagonistic reaction.

[0005] The present invention is achieved through the following technical solutions:

[0006] A blood sugar lowering complex including quercetin and 3-O-methylquercetin.

[0007] The present invention provides a hypoglycemic compound, which can reduce blood sugar by reasonable compounding and avoid antagonistic reaction between two components.

[0008] Furthermore, the mass ratio of quercetin to 3-O-methylquercetin is 6:4 to 2:8. The mass ratio of quercetin to 3-O-methylquercetin is limited to achieve a synergistic effect, thereby reducing the dosage, reducing side effects, and lowering blood sugar.

[0009] Furthermore, the hypoglycemic compound further comprises at least one of a carrier, a solvent, a diluent and an excipient for carrying quercetin and 3-O-methylquercetin.

[0010] The present invention also provides a pharmaceutical dosage form, including any one of powder, granule, capsule, injection, oral liquid, tablet, dripping pill or granule.

[0011] The present invention also provides an application of the hypoglycemic compound in the field of inhibiting α-glucosidase.

[0012] The present invention also provides an α-glucosidase inhibitor comprising the above-mentioned blood sugar-lowering compound.

[0013] For better understanding and implementation, the present invention is described in detail below with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Schematic diagram of the interaction between 3-O-methylquercetin and α-glucosidase.

[0015] Figure 2 Schematic diagram of the interaction between quercetin and α-glucosidase.

[0016] Figure 3 This is the inhibition curve of quercetin on α-glucosidase.

[0017] Figure 4 This is the inhibition curve of 3-O-methylquercetin on α-glucosidase.

[0018] Figure 5 This is an isobol analysis of the interaction of quercetin and 3-O-methylquercetin combination at different ratios with α-glucosidase. DETAILED DESCRIPTION

[0019] The following is a further detailed description of the embodiments of the present invention in conjunction with the accompanying drawings and examples. It should be understood that the specific embodiments described herein are merely illustrative of the embodiments of the present invention and are not intended to limit the embodiments of the present invention. It should also be noted that, for ease of description, the accompanying drawings only illustrate portions of the embodiments of the present invention, rather than all structures.

[0020] Furthermore, the terms "first," "second," "third," etc., in the specification and claims are used solely for descriptive purposes to distinguish between identical technical features. They are not to be construed as indicating or implying relative importance, or as implicitly specifying the number of technical features, nor do they necessarily describe a sequential or chronological order. The terms are interchangeable where appropriate. Thus, a feature specified as "first" or "second" may explicitly or implicitly include at least one of those features.

[0021] Similarly, the terms "fixed" and "connected" used in the specification and claims should not be construed as limited to direct connections. Thus, the expression "device A is connected to device B" should not be limited to devices or systems in which device A is directly connected to device B. Rather, it means that a path exists between device A and device B, which may include other devices or tools.

[0022] Example 1

[0023] This embodiment provides a hypoglycemic compound comprising quercetin and 3-O-methylquercetin, wherein the mass ratio of quercetin to 3-O-methylquercetin is 6:4 to 2:8. The mass ratio of quercetin to 3-O-methylquercetin is limited to achieve a synergistic effect, thereby reducing the dosage and side effects while lowering blood sugar.

[0024] The hypoglycemic compound further comprises at least one of a carrier, a solvent, a diluent and an excipient for carrying quercetin and 3-O-methylquercetin, and can be prepared into corresponding powders, granules, capsules, injections, oral solutions or tablets according to different requirements.

[0025] The hypoglycemic complex provided in this embodiment can inhibit the activity of α-glucosidase, thereby achieving the effect of lowering blood sugar. By rationally compounding and combining drugs, a synergistic effect is achieved, the dosage of each component when used alone is reduced, and the inhibitory activity against α-glucosidase is improved. Low-dose combination therapy can effectively reduce the occurrence of drug resistance in the human body.

[0026] Quercetin, molecular formula C 15 H 10 O7; molecular weight: 302.24; CAS registration number: 117-39-5, structural formula:

[0027]

[0028] 3-O-Methylquercetin, molecular formula C 16 H 12 O7; molecular weight: 316.26; CAS registration number: 1486-70-0, structural formula:

[0029]

[0030] Figure 1 This is a schematic diagram of the interaction between 3-O-methylquercetin and α-glucosidase. Figure 2 This is a schematic diagram of the interaction between quercetin and α-glucosidase, please refer to Figure 1-2When 3-O-methylquercetin binds to α-glucosidase, it is mainly surrounded by hydrophobic amino acid residues such as SER156, PHE157, LEU176, LEU218, HIS239, PRO240, ASN241, PHE310, and ARG312. In addition, three hydrogen bonds are formed between 3-O-methylquercetin and α-glucosidase residues LYS155, SER308, and SER309, with bond lengths of When quercetin binds to α-glucosidase, it is mainly surrounded by amino acid residues such as ASN317, GLU429, PHE314, ILE419, GLU422, HIS423, ALA418, GLY160, LYS156, and ASN235, and forms a hydrogen bond with amino acid ASP233, with an average hydrogen bond length of 3-O-methylquercetin and quercetin bind to different amino acid residues of α-glucosidase, and their inhibitory activities on α-glucosidase are also different. Through reasonable compounding, the inhibitory activities on different amino acid residues of α-glucosidase can be controlled to avoid antagonism and achieve synergistic effect.

[0031] The in vitro inhibition test of α-glucosidase activity is as follows:

[0032] The hypoglycemic complex or control sample was dissolved in dimethyl sulfoxide (DMSO) to prepare a 10 mg / mL stock solution. The stock solution was diluted with PBS, and the DMSO content after dilution was less than 5%. A gradient concentration was set, and 100 μL was taken from the diluted solution of each gradient concentration as the sample to be tested. 40 μL of α-glucosidase (0.25 U / mL) was added to the sample to be tested, and the sample was placed in a 37°C environment to react for 10 minutes; then 60 μL of 5 mmol / mL substrate p-nitrophenyl-α-D-pyranoglucoside (pNPG) was added, and the sample was placed in a 37°C environment to react for 15 minutes, and then measured at a wavelength of 405 nm using a microplate reader to obtain the OD value. The inhibition rate of α-glucosidase activity at each gradient concentration was calculated based on the OD value, and the inhibition rate = [1-(OD 样品 -OD 样品空白 ) / (OD 阴性对照 -OD 空白 )] × 100%. The half inhibitory concentration (IC50) was calculated based on the inhibition rate of α-glucosidase activity at each gradient concentration. 50 The calculation and statistical process were performed using SPSS 20.0.

[0033] Quercetin, 3-O-methylquercetin and acarbose were used as test objects. The inhibitory activity of each test object on α-glucosidase was tested by the above method. The half inhibitory concentration IC 50 Value representation.

[0034] Specifically, the experimental group settings are as follows:

[0035] Sample group: 100 μL test sample + 40 μL α-glucosidase + 60 μL pNPG;

[0036] Sample blank group: 100 μL test sample + 40 μL PBS + 60 μL pNPG;

[0037] Positive control group: 100 μL acarbose + 40 μL α-glucosidase + 60 μL pNPG;

[0038] Negative control group: 2 μL DMSO + 98 μL PBS + 40 μL α-glucosidase + 60 μL pNPG;

[0039] Blank control group: 2 μL DMSO + 98 μL PBS + 40 μL PBS + 60 μL pNPG.

[0040] Principles for setting a concentration gradient: Based on the initial concentration, perform two-fold dilutions in sequence. For example, for a 6:4 ratio of quercetin and 3-O-methylquercetin, the initial concentration of the hypoglycemic complex is set at 10 μg / mL, with an initial concentration of 6 μg / mL for quercetin and 4 μg / mL for 3-O-methylquercetin. Based on the initial concentration of 10 μg / mL, perform two-fold dilutions in sequence to 10 μg / mL, 5 μg / mL, 2.5 μg / mL, and 1.25 μg / mL. Similarly, for a 5:5 ratio of quercetin and 3-O-methylquercetin, the initial concentration is set at 10 μg / mL and then diluted two-fold to form a concentration gradient. This principle is also followed for other compositions.

[0041] The drug interactions of hypoglycemic compounds were determined by isobol method and the interaction index was calculated.

[0042] IC 50 The calculation formula of the theoretical value is: IC 50add =IC 50A / (P1+R*P2), where R is the potency ratio of compounds A and B when acting alone, i.e. R=IC 50A / IC 50B , P1 is the proportion of compound A in the hypoglycemic complex, and P2 is the proportion of compound B in the hypoglycemic complex. Synergistic rate of hypoglycemic complex (%) = (IC 50add -IC 50mix ) / IC 50add *100.

[0043] Interaction index (γ) is the interaction index value = IC 50Amix / IC 50A +IC 50Bmix / IC 50B , among which, IC 50Amix IC is the value in the coordinate system corresponding to compound A when the hypoglycemic compound reaches the corresponding inhibition rate. 50Bmix is the value in the coordinate system corresponding to compound B when the hypoglycemic compound reaches the corresponding inhibition rate. When γ>1, it shows antagonism, when γ=1, it shows additive effect, and when γ<1, it shows additive effect.

[0044] The combination of quercetin and 3-O-methylquercetin was used as the test object to test the inhibitory activity of each test object on α-glucosidase, and the half inhibitory concentration IC 50 Value representation.

[0045] The specific test objects are: quercetin and 3-O-methylquercetin (6:4), quercetin and 3-O-methylquercetin (5:5), quercetin and 3-O-methylquercetin (4:6) and quercetin and 3-O-methylquercetin (2:8), and the above ratios are all mass ratios.

[0046] The concentration gradient of each experimental object was set as follows:

[0047] Quercetin and 3-O-methylquercetin (6:4): (6+4) μg / mL, (3+2) μg / mL, (1.5+1) μg / mL, (0.75+0.5) μg / mL;

[0048] Quercetin and 3-O-methylquercetin (5:5): (5+5) μg / mL, (2.5+2.5) μg / mL, (1.25+1.25) μg / mL, (0.625+0.625) μg / mL;

[0049] Quercetin and 3-O-methylquercetin (4:6): (4+6) μg / mL, (2+3) μg / mL, (1+1.5) μg / mL, (0.5+0.75) μg / mL;

[0050] Quercetin and 3-O-methylquercetin (2:8): (2+8) μg / mL, (1+4) μg / mL, (0.5+2) μg / mL, (0.25+1) μg / mL.

[0051] Table 1 is the drug interaction table of hypoglycemic compound combination, IC 50MIX is the experimental value of the composition, IC 50addRefer to Table 1, using single-component quercetin and single-component 3-O-methylquercetin as comparison, the IC value of 10μg / mL quercetin is 50 The IC value of 3-O-methylquercetin was 2.28±0.29μg / mL and 10mg / mL 50 The value was 4.22±0.84μg / mL. In the hypoglycemic compound, quercetin and 3-O-methylquercetin were used together, the total concentration of the hypoglycemic compound was 10μg / mL, and the IC values ​​of quercetin and 3-O-methylquercetin were 6:4, 5:5, 4:6, and 2:8. 50 The values ​​were 2.05±0.16, 2.55±0.47, 2.43±0.12 and 2.82±0.52 μg / mL, respectively. The interaction index (γ) was less than 1, which were 0.73, 0.86, 0.77 and 0.78, respectively. The synergistic rates were 26.59%, 14.01%, 22.90% and 21.86%, respectively, all of which were positive, showing strong synergistic effects.

[0052] Figure 3 This is the inhibition curve of quercetin on α-glucosidase. Figure 3 Quercetin has a good inhibitory effect on α-glucosidase, and its inhibitory effect is better than that of the commonly used acarbose inhibitor. Figure 4 This is the inhibition curve of 3-O-methylquercetin on α-glucosidase, please refer to Figure 4 3-O-methylquercetin has a good inhibitory effect on α-glucosidase, and its inhibitory effect is better than that of the commonly used acarbose inhibitor.

[0053] Figure 5 This is an isobol analysis of the interaction of quercetin and 3-O-methylquercetin combination with α-glucosidase at different ratios. Please also refer to Figure 5 As shown in Table 1, when quercetin and 3-O-methylquercetin were used in combination at ratios of 6:4, 5:5, 4:6, and 2:8, the interaction index (γ) was less than 1, reflecting a synergistic effect rather than a simple additive effect. The same hypoglycemic effect as quercetin or 3-O-methylquercetin could be achieved, and the amounts of quercetin and 3-O-methylquercetin in the hypoglycemic compounds were reduced compared with those used alone.

[0054] Therefore, the hypoglycemic complex provided in this embodiment can achieve the same hypoglycemic effect as single-component quercetin and single-component 3-O-methylquercetin at a lower dosage. The combined use significantly improves the inhibitory activity against α-glucosidase. The use of the hypoglycemic complex can reduce the dosage of quercetin and 3-O-methylquercetin to alleviate the side effects of the drugs on the human body and control blood sugar.

[0055] Table 1 Table of drug interactions of combined use of hypoglycemic compounds

[0056]

[0057] Example 2

[0058] This embodiment provides an application of a hypoglycemic compound in the field of inhibiting α-glucosidase. The hypoglycemic compound includes quercetin and 3-O-methylquercetin, and the mass ratio of quercetin to 3-O-methylquercetin is 6:4 to 2:8. The compound can be prepared into corresponding powders, granules, capsules, injections, oral solutions or tablets according to different needs by adding pharmaceutically or food-acceptable carriers, solvents, diluents, excipients or other media, drugs or health products, to prepare hypoglycemic drugs or health products.

[0059] Example 3

[0060] This embodiment provides a hypoglycemic compound, comprising quercetin and 3-O-methylquercetin, wherein the mass ratio of quercetin to 3-O-methylquercetin is 6:4, wherein the concentration of quercetin is 6 μg / mL, and the concentration of 3-O-methylquercetin is 4 μg / mL.

[0061] Example 4

[0062] This embodiment provides a hypoglycemic compound, comprising quercetin and 3-O-methylquercetin, wherein the mass ratio of quercetin to 3-O-methylquercetin is 5:5, wherein the concentration of quercetin is 5 μg / mL, and the concentration of 3-O-methylquercetin is 5 μg / mL.

[0063] Example 5

[0064] This embodiment provides a hypoglycemic compound, comprising quercetin and 3-O-methylquercetin, wherein the mass ratio of quercetin to 3-O-methylquercetin is 4:6, wherein the concentration of quercetin is 4 μg / mL, and the concentration of 3-O-methylquercetin is 6 μg / mL.

[0065] Example 6

[0066] This embodiment provides a hypoglycemic compound, comprising quercetin and 3-O-methylquercetin, wherein the mass ratio of quercetin to 3-O-methylquercetin is 2:8, wherein the concentration of quercetin is 2 μg / mL, and the concentration of 3-O-methylquercetin is 8 μg / mL.

[0067] The present invention is not limited to the above-mentioned embodiments. If various changes or modifications of the present invention do not depart from the spirit and scope of the present invention, and if these changes and modifications fall within the scope of the claims of the present invention and equivalent technologies, the present invention is also intended to include these changes and modifications.

Claims

1. A hypoglycemic compound, characterized by: including quercetin and 3-O-methylquercetin; The mass ratio of quercetin to 3-O-methylquercetin is 6:

4.

2. The hypoglycemic compound according to claim 1, characterized in that: Also included are carriers for carrying quercetin and 3-O-methylquercetin.

3. A pharmaceutical dosage form comprising the hypoglycemic compound according to any one of claims 1 to 2, characterized in that: Including any one of powder, granules, capsules, injections, oral solutions, tablets or pills.

4. Use of the hypoglycemic compound according to any one of claims 1 to 2 in the preparation of a medicament having a synergistic hypoglycemic effect, characterized in that: The hypoglycemic compound inhibits the activity of α-glucosidase, achieves a synergistic effect, and reduces the dosage of each component when used alone to achieve the improvement of the inhibitory activity on α-glucosidase.

5. An α-glucosidase inhibitor, characterized in that: The invention comprises the blood sugar lowering compound according to any one of claims 1 to 2.

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