A coumarin derivative containing a thiazolidinedione structure, its preparation method and application

Abstract

This invention discloses a coumarin derivative containing a thiazolidinedione structure, its preparation method, and its application, belonging to the field of medicinal chemistry. The coumarin derivative containing the thiazolidinedione structure of this invention has the structure shown in the following formula: where R represents a substituted or unsubstituted phenyl group, and the substitution is optional. This invention designs and synthesizes a series of coumarin derivatives containing a thiazolidinedione structure. These coumarin derivatives exhibit strong α-glucosidase inhibitory activity, with an IC50 value of [missing information]. 50 The lowest value is 5.12±0.98 μM, which is 34 to 128 times that of acarbose, and it can be used to treat and / or prevent diabetes. This invention also provides a method for preparing and using coumarin derivatives containing a thiazolidinedione structure.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and in particular to a coumarin derivative containing a thiazolidinedione structure, its preparation method, and its application. Background Technology

[0002] Currently, diabetes has become a global problem, seriously endangering human health. Despite increased health awareness, the incidence of diabetes continues to rise. If blood sugar levels are not controlled effectively and promptly, it can induce a series of functional disorders and damages in the body, seriously threatening the health and safety of patients. Diabetes is divided into type 1 diabetes and type 2 diabetes. Type 1 diabetes, also known as insulin-dependent diabetes mellitus (IDDM), accounts for about 10% of diabetes patients. It is mainly caused by severe damage to the pancreatic β cells that secrete insulin, resulting in insufficient endogenous insulin secretion, and has a certain degree of heritability. Type 2 diabetes, also known as non-insulin-dependent diabetes mellitus (NIDDM), is a chronic metabolic disease caused by insulin dysfunction or insulin resistance. It is mainly caused by the interaction of genetic and environmental factors, such as gene mutations and dietary habits, and accounts for more than 90% of cases.

[0003] Currently, there are various oral hypoglycemic drugs on the market, which act on different targets through different pathways. Sulfonylureas (glibenclamide, glimepiride, and tolbutamide) are insulin secretagogues that can activate voltage-gated calcium channel blockers. 2+The insulin-producing pathway stimulates pancreatic β-cells to release insulin. Biguanides, such as metformin, are considered first-line treatments for diabetes. Their mechanism of action involves inhibiting hepatic gluconeogenesis and hepatic glucose output, thereby lowering blood glucose levels. Sodium-glucose transporter 2 (SGLT2) inhibitors, such as dapagliflozin, inhibit glucose reabsorption, thus controlling blood glucose and reducing calorie intake. Another method for maintaining blood glucose levels is inhibiting the hydrolysis of α-glucosidase, thereby lowering postprandial blood glucose. α-glucosidase aids digestion by hydrolyzing starch and carbohydrates into glucose and monosaccharides in the small intestine. α-glucosidase inhibitors competitively and reversibly inhibit this intestinal enzyme, preventing the hydrolysis of the non-reducing ends of oligosaccharides and reducing the release of α-glucose. This hinders carbohydrate digestion and absorption, thereby reducing insulin requirements. Furthermore, α-glucosidase inhibitors can lead to a long-term, sustained increase in endogenous GLP-1, thereby stimulating insulin secretion and reducing glucagon secretion, thus controlling postprandial hyperglycemia. Currently available alpha-glucosidase inhibitors include acarbose, voglibose, and miglitol. However, long-term use of these drugs may cause side effects such as diarrhea, abdominal and gastrointestinal discomfort. Therefore, developing drugs with good efficacy and fewer side effects is of great significance. In recent years, many new studies on alpha-glucosidase inhibitors have been conducted, which may contribute to the discovery of new alpha-glucosidase inhibitors in the future.

[0004] Coumarins are naturally occurring organic compounds that have become a research hotspot due to their wide range of biological activities, such as antioxidant, anti-inflammatory, antibacterial, and anticancer effects. However, research on coumarin compounds as α-glucosidase inhibitors is limited. Therefore, combining the coumarin skeleton with other biologically active structures may yield new and effective α-glucosidase inhibitors. Thiazolidinediones are compounds with broad biological activities and have been shown to have significant effects in antibacterial, anti-tuberculosis, and anticancer fields. According to existing literature, 5-benzylthiazolidin-2,4-dione IC 50 =98.88±1.11μM, IC50 of coumarin analog 3-acetylcoumarin 50 =1.5×10 5 μM.

[0005] Currently, no coumarin derivatives containing thiazolidinedione structures have been publicly disclosed. Therefore, developing new derivatives with α-glucosidase inhibitory activity remains of great significance. Summary of the Invention

[0006] The present invention aims to at least solve one of the technical problems existing in the prior art. To this end, the present invention proposes a coumarin derivative containing a thiazolidinedione structure, which can effectively inhibit α-glucosidase, thereby being used for the treatment and / or prevention of diabetes.

[0007] The present invention also provides a method for preparing the above-mentioned coumarin derivatives containing a thiazolidinedione structure.

[0008] The present invention also provides applications of the above-mentioned coumarin derivatives containing a thiazolidinedione structure.

[0009] According to a first aspect of the present invention, a coumarin derivative containing a thiazolidinedione structure is provided, said coumarin derivative having the structural formula shown below:

[0010]

[0011] In the formula, R is selected from substituted or unsubstituted phenyl groups, and the substitution is optional.

[0012] The coumarin derivatives containing a thiazolidinedione structure according to the first aspect of the present invention have at least the following beneficial effects:

[0013] This invention uses the coumarin structure as a core to design and synthesize a series of coumarin derivatives containing thiazolidinedione structures. Among them, the coumarin derivatives with Br substitution at the meta position exhibit strong α-glucosidase inhibitory activity at C15, with an IC5 value of [missing information]. 50 The highest value was 5.12 ± 0.98 μM, which is 128 times that of acarbose. Compared with single thiazolidinedione derivatives and single coumarin derivatives, the compounds of this invention exhibit significant α-glucosidase inhibitory activity. Kinetic studies show that C15 is a reversible, non-competitive inhibitor and can be used as an α-glucosidase inhibitor for the treatment or prevention of diabetes.

[0014] According to some embodiments of the present invention, the substitution is a single substitution or multiple substitutions.

[0015] According to some embodiments of the present invention, the substituent is selected from at least one of halogen, methyl, methoxy, and hydroxyl.

[0016] According to some embodiments of the present invention, the R is selected from the following groups:

[0017]

[0018] According to a second aspect of the present invention, a method for preparing a coumarin derivative containing a thiazolidinedione structure is provided, comprising the following steps:

[0019] The coumarin derivative containing the thiazolidinedione structure was obtained by reacting compound 2 and compound 5 containing the R group as raw materials;

[0020] The structures of compounds 2 and 5 are as follows:

[0021]

[0022] According to some embodiments of the present invention, the molar ratio of compound 2 to compound 5 is (1.2 to 1.5):1. For example, it can be about 1.2:1, 1.3:1, 1.4:1, or 1.5:1.

[0023] According to some embodiments of the present invention, the solvent for the reaction is tetrahydrofuran (THF) or dimethylformamide (DMF).

[0024] According to some embodiments of the present invention, the reaction temperature is 20–30°C. For example, it can be approximately 20°C, 25°C, or 30°C.

[0025] According to some embodiments of the present invention, the reaction time is 1 to 3 days. For example, it can be approximately 1 day, 2 days, or 3 days.

[0026] According to some embodiments of the present invention, the reaction stirring speed is approximately 400–600 rpm. For example, it can be approximately 400 rpm, 500 rpm, or 600 rpm.

[0027] According to some embodiments of the present invention, the post-reaction processing step includes: adding an appropriate amount of water to the reaction system, producing a solid, and washing and filtering with water to obtain the final product.

[0028] According to some embodiments of the present invention, the preparation method of compound 2 includes the following steps:

[0029] Compound 2 was obtained by reacting 7-amino-4-methylcoumarin (compound 1) with bromoacetyl bromide in the presence of a base at room temperature.

[0030] The structural formula of compound 1 is as follows:

[0031]

[0032] According to some embodiments of the present invention, the solvent for the reaction is dichloromethane (DCM).

[0033] According to some embodiments of the present invention, the base is triethylamine (TEA).

[0034] According to some embodiments of the present invention, the molar ratio of compound 1 to bromoacetyl bromide is 1:(1.2 to 1.4). For example, it can be about 1:1.2, 1:1.3, or 1:1.4.

[0035] According to some embodiments of the present invention, the molar ratio of compound 1 to TEA is 1:1.5.

[0036] According to some embodiments of the present invention, the reaction is carried out in an inert atmosphere.

[0037] According to some embodiments of the present invention, the inert atmosphere includes at least one of nitrogen, argon and helium.

[0038] Preferably, the inert atmosphere is nitrogen (N2).

[0039] According to some embodiments of the present invention, the preparation process of compound 5 includes the following steps:

[0040] Compound 5 was obtained by reacting 2,4-thiazolidinedione (compound 4) with benzaldehyde containing an R-group in the presence of a base at room temperature.

[0041] The structural formula of compound 4 is as follows:

[0042]

[0043] According to some embodiments of the present invention, the molar ratio of compound 4 to benzaldehyde substituted with an R group is (1 to 1.2):1. For example, it can be about 1:1, 1.1:1, or 1.2:1.

[0044] According to some embodiments of the present invention, the solvent for the reaction is anhydrous ethanol (EtOH).

[0045] According to some embodiments of the present invention, the alkali is sodium hydroxide (NaOH).

[0046] According to some embodiments of the present invention, the post-reaction processing step includes: quenching with water, washing with n-hexane, and filtering to obtain the product.

[0047] According to a third aspect of the present invention, an α-glucosidase inhibitor is proposed comprising the above-described coumarin derivatives containing a thiazolidinedione structure.

[0048] According to a fourth aspect of the present invention, the use of the above-mentioned coumarin derivatives containing a thiazolidinedione structure in the preparation of products for the prevention and / or treatment of diabetes is proposed.

[0049] According to some embodiments of the present invention, the product includes at least one of a drug and a health product.

[0050] According to a fifth aspect of the invention, a pharmaceutical composition is provided comprising the above-described coumarin derivatives containing a thiazolidinedione structure and / or their pharmaceutically acceptable salts.

[0051] According to some embodiments of the present invention, the dosage form of the drug is tablets, capsules, oral liquids or injections.

[0052] The structural formula appears in this invention Indicates the linking site of a functional group.

[0053] The term "pharmaceutically acceptable" as used in this invention refers to a substance that is acceptable from a toxicological point of view for pharmaceutical use and will not adversely interact with the active ingredient.

[0054] Other features and advantages of the invention will be set forth in the description which follows, and will be apparent in part from the description, or may be learned by practicing the invention. Attached Figure Description

[0055] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0056] Figure 1 Example 2 of the present invention provides a 50% inhibitory concentration (50%) diagram of a coumarin derivative C15 containing a thiazolidinedione structure as an α-glucosidase inhibitor against α-glucosidase in vitro.

[0057] Figure 2 The image shows the in vitro enzyme kinetics of α-glucosidase by C15, a coumarin derivative containing a thiazolidinedione structure, as an α-glucosidase inhibitor in Example 3 of this invention.

[0058] Figure 3 This is a diagram showing the substrate kinetics of α-glucosidase in vitro using C15, a coumarin derivative containing a thiazolidinedione structure, as an α-glucosidase inhibitor in Example 4 of the present invention.

[0059] Figure 4 This is a graph showing the cytotoxicity test results of LO2 cells using C15, a coumarin derivative containing a thiazolidinedione structure, as an α-glucosidase inhibitor in Example 5 of the present invention.

[0060] Figure 5 This is a diagram showing the toxicity test results of C15, a coumarin derivative containing a thiazolidinedione structure, as an α-glucosidase inhibitor on 293 cells in Example 5 of the present invention. Detailed Implementation

[0061] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0062] In the description of this invention, the use of terms such as "first," "second," etc., is for the purpose of distinguishing technical features only and should not be construed as indicating or implying relative importance, or implicitly indicating the number of technical features indicated, or implicitly indicating the order of the technical features indicated.

[0063] When a numerical range is disclosed herein, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0064] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of the present invention.

[0065] Unless otherwise specified, the reagents, methods and equipment used in this invention are all conventional reagents, methods and equipment in this technical field.

[0066] Reagents used in the embodiments of this invention:

[0067] CAS No.: 26093-31-2;

[0068] CAS number for bromoacetyl bromide: 598-21-0;

[0069] Triethylamine CAS No.: 121-44-8;

[0070] Sodium hydroxide CAS number: 1310-73-2;

[0071] 2,4-Thiazolidinedione CAS No.: 2295-31-0;

[0072] Example 1

[0073] This embodiment prepared a series of coumarin derivatives containing thiazolidinedione structures, and the preparation route is as follows:

[0074]

[0075] The preparation method includes the following specific steps:

[0076] 7-Amino-4-methylcoumarin (1 mmol), bromoacetyl bromide (1.3 mmol), and triethylamine (1.5 mmol) were reacted in anhydrous DCM solution for 12 hours. After the reaction was complete, the solid was collected and then washed with 100 mL of water to give compound (2).

[0077] The substituted benzaldehyde (1 mmol), thiazolidinedione (1.2 mmol), and sodium bicarbonate (3 mmol) were reacted in anhydrous ethanol solution (10 mL). After 4 hours, the solid was extracted, filtered, washed with n-hexane (50 mL), and the solid was collected to give compound (5).

[0078] Compounds (2, 1.2 mmol), compound (5, 1 mmol), and triethylamine (3 mmol) were stirred in DMF solution at 500 rpm for 3 days. After the reaction, the mixture was cooled with ice. The resulting solids were collected and washed with 200 mL of water to give a series of products C (named C1–C20).

[0079] In the formula, R in C1 to C20 is shown in Table 1 below:

[0080] Table 1. Structural formulas of the R group

[0081]

[0082]

[0083]

[0084] The structures of C1–C20 compounds were characterized by NMR, MS, and melting point analysis. The following are the morphology, yield, NMR, and mass spectrometry results for each compound:

[0085] (Z)-2-(5-benzylidene-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamid e(C1).Yellow solid; yield 51%; mp 302.5-303.1℃; 1H NMR(500MHz,DMSO)δ10.87(s,1H,NH),8.01(s,1H,CH=),7.76(d,J=10.0H z,1H,Ar-H),7.69(s,1H,Ar-H),7.67(s,2H,Ar-2H),7.58(d,J=5.0Hz,1H ,Ar-H),7.56(s,1H,Ar-H),7.54(d,J=5.0Hz,1H,Ar-H),7.49(d,J=10.0H z,1H,Ar-H),6.30-6.28(m,1H,Ar-H),4.59(s,2H,CH2),2.41(s,3H,CH3); 13 C NMR(126MHz,DMSO)δ167.43(C=O),165.55(C=O),164.94(C=O),160.24(C=O),153.91( CH),153.36(Ar-C),141.85(Ar-C),134.09(Ar-C),133.11(Ar-C),131.18(Ar-C),130 .53(Ar-2C),129.74(Ar-2C),126.46(Ar-C),121.15(Ar-C),115.74(Ar-C),115.46(Ar-C -C),112.86(Ar-C),106.04(Ar-C),44.48(CH2),18.28(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 16 N2O5S + :421.0852;found:421.0853.

[0086] (Z)-N-(4-methyl-2-oxo-2H-chromen-7-yl)-2-(5-(2-methylbenzylidene)-2,4-dioxothiazolidin-3-yl)acetamide(C2). 1H NMR(500MHz,DMSO)δ10.87(s,1H,NH),8.08(s,1H,CH=),7.74(d,J=10.0Hz,1H,Ar-H),7.66(s,1H,Ar-H),7.48(s,1H,Ar-H),7.47(s,1H,Ar-H),7.40(d,J=5.0Hz,1H,Ar-H),7.38(s,1H,Ar-H),7.36(s,1H,Ar-H),6.28(s,1H,Ar-H),4.58(s,2H,CH2),2.41(s,3H,CH3),2.39(s,3H,CH3); 13 C NMR(126MHz,DMSO)δ167.56(C=O),165.18(C=O),164.85(C=O),160.14(C=O),153.81(CH=),153.26(Ar-C),141.75(Ar-C),139.09(Ar-C),132.15(Ar-C),131.90(Ar-C),131.26(Ar-C),130.89(Ar-C),127.45(Ar-C),126.94(Ar-C),126.37(Ar-C),122.78(Ar-C),115.64(Ar-C),115.35(Ar-C),112.76(Ar-C),105.93(Ar-C),44.32(CH2),19.58(CH3),18.18(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 23 H 18 N2O5S + :435.1009;found:435.1013.

[0087] (Z)-N-(4-methyl-2-oxo-2H-chromen-7-yl)-2-(5-(3-methylbenzylidene)-2,4-dioxothiazolidin-3-yl)acetamide(C3).Yellow solid;yield 47%;mp.299.0-299.4℃; 1H NMR(500MHz,DMSO)δ10.87(s,1H,NH),7.95(s,1H,CH=),7.75(d,J=10.0Hz,1H,Ar-H),7.66(s,1H,Ar-H),7.49(s,1H,Ar-H),7.46(s,1H,Ar-H),7.45(s,1H,Ar-H),7.35(d,J=5.0Hz,1H,Ar-H),6.28(s,1H,Ar-H),4.58(s,2H,CH2),2.40(s,3H,CH3),2.38(s,3H,CH3); 13 CNMR(126MHz,DMSO)δ167.24(C=O),165.36(C=O),164.77(C=O),160.03(C=O),153.73(CH=),153.17(Ar-C),141.67(Ar-C),138.93(Ar-C),133.98(Ar-C),132.91(Ar-C),131.73(Ar-C),130.92(Ar-C),129.45(Ar-C),127.37(Ar-C),126.30(Ar-C),120.77(Ar-C),115.55(Ar-C),115.27(Ar-C),112.67(Ar-C),105.84(Ar-C),44.28(CH2),21.02(CH3),18.09(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 23 H 18 N2O5S + :435.1009;found:435.1013.

[0088] (Z)-N-(4-methyl-2-oxo-2H-chromen-7-yl)-2-(5-(4-methylbenzylidene)-2,4-dioxothiazolidin-3-yl)acetamide(C4).White solid;yield 45%;mp 302.5-303.4℃; 1H NMR(500MHz,DMSO)δ10.86(s,1H,NH),7.96(s,1H,=CH),7.73(d,J=5.0Hz,1H,Ar-H),7.66(s,1H,Ar-H),7.56(s,1H,Ar-H),7.55(s,1H,Ar-H),7.48(d,J=5.0Hz,1H,Ar-H),7.38(s,1H,Ar-H),7.37(s,1H,Ar-H),6.27(s,1H,Ar-H),4.58(s,2H,CH2),2.39(s,3H,CH3),2.37(s,3H,CH3); 13 C NMR(126MHz,DMSO)δ167.16(C=O),165.32(C=O),164.68(C=O),159.94(C=O),153.63(=CH),153.05(Ar-C),141.58(Ar-C),141.28(Ar-C),133.86(Ar-C),130.33(Ar-2C),130.07(Ar-3C),126.17(Ar-C),119.60(Ar-C),115.44(Ar-C),115.16(Ar-C),112.57(Ar-C),105.74(Ar-C),44.16(CH2),21.14(CH3),17.99(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 23 H 18 N2O5S + :435.1009;found:435.1013.

[0089] (Z)-2-(5-(2-methoxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C5).White solid;yield 53%;mp 284.7-285.1℃; 1H NMR(500MHz,DMSO)δ10.87(s,1H,NH),7.99(s,1H,=CH),7.75(d,J=9.1Hz,1H,Ar-H),7.67(s,1H,Ar-H),7.50(d,J=6.7Hz,1H,Ar-H),7.47(s,1H,Ar-H),7.24(s,1H,Ar-H),7.23(s,1H,Ar-H),7.11(d,J=8.0Hz,1H,Ar-H),6.29(s,1H,Ar-H),4.58(s,2H,CH2),3.82(s,3H,OCH3),2.40(s,3H,CH3); 13 C NMR(126MHz,DMSO)δ168.02(C=O),166.00(C=O),165.32(C=O),160.58(C=O),158.70(=CH),154.25(Ar-C),153.70(Ar-C),142.20(Ar-C),133.59(Ar-C),129.72(Ar-C),129.37(Ar-C),126.80(Ar-C),121.82(Ar-C),121.72(Ar-C),121.57(Ar-C),116.06(Ar-C),115.79(Ar-C),113.18(Ar-C),112.63(Ar-C),106.36(Ar-C),56.44(CH2),44.74(OCH3),18.61(CH3);HRMS(ESI-MS)m / z:[M+H] + calcdfor C 23 H 18 N2O6S + :451.0958;found:451.0960.

[0090] (Z)-2-(5-(3-methoxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C6).Yellow solid;yield 57%;mp 284.5-284.7℃; 1H NMR(500MHz,DMSO)δ10.86(s,1H,NH),7.96(s,1H,=CH),7.75(d,J=8.7Hz,1H,Ar-H),7.67(s,1H,Ar-H),7.65(s,1H,Ar-H),7.63(s,1H,Ar-H),7.48(d,J=8.6Hz,1H,Ar-H),7.13(d,J=8.4Hz,2H,Ar-2H),6.28(s,1H,Ar-H),4.57(s,2H,CH2),3.84(s,3H,OCH3),2.40(s,3H,CH3); 13 C NMR(126MHz,DMSO)δ167.56(C=O),165.69(C=O),165.13(C=O),160.44(C=O),160.16(=CH),154.10(Ar-C),153.57(Ar-C),142.03(Ar-C),134.65(Ar-C),134.25(Ar-C),131.05(Ar-C),126.66(Ar-C),122.49(Ar-C),121.71(Ar-C),117.24(Ar-C),116.04(Ar-C),115.93(Ar-C),115.66(Ar-C),113.04(Ar-C),106.23(Ar-C),55.81(CH2),44.65(OCH3),18.47(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 23 H 18 N2O6S + :451.0958;found:451.0960.

[0091] (Z)-2-(5-(4-methoxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C7).Yellow solid;yield 41%;mp 304.3-304.9℃; 1H NMR(500MHz,DMSO)δ10.86(s,1H,NH),7.96(s,1H,=CH),7.75(d,J=10.0Hz,1H,Ar-H),7.67(s,1H,Ar-H),7.65(s,1H,Ar-H),7.63(s,1H,Ar-H),7.48(d,J=10.0Hz,1H,Ar-H),7.13(d,J=10.0Hz,2H,Ar-2H),6.28(s,1H,Ar-H),4.57(s,2H,CH2),3.84(s,3H,OCH3),2.40(s,3H,CH3); 13 C NMR(126MHz,DMSO)δ167.65(C=O),165.82(C=O),165.18(C=O),161.80(C=O),160.39(=CH),154.07(Ar-C),153.52(Ar-C),142.03(Ar-C),134.23(Ar-C),132.87(Ar-2C),126.64(Ar-C),125.74(Ar-C),117.97(Ar-C),115.88(Ar-C),115.61(Ar-C),115.50(Ar-2C),113.00(Ar-C),106.18(Ar-C),56.00(CH2),44.56(OCH3),18.43(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 23 H 18 N2O6S + :451.0958;found:451.0963.

[0092] (Z)-2-(5-(2-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C8).White solid;yield 47%;mp 278.6-278.9℃; 1H NMR(500MHz,DMSO)δ10.89(s,1H,NH),8.10(s,1H,=CH),7.75(d,J=10.0Hz,1H,Ar-H),7.67(s,2H,Ar-2H),7.64(s,1H,Ar-H),7.56(s,2H,Ar-H),7.48(d,J=10.0Hz,1H,Ar-H),6.29(s,1H,Ar-H),4.60(s,2H,CH2),2.40(s,3H,CH3); 13 C NMR(126MHz,DMSO)δ167.32(C=O),165.32(C=O),164.99(C=O),160.38(C=O),154.06(=CH),153.48(Ar-C),141.98(Ar-C),135.00(Ar-C),132.72(Ar-C),131.26(Ar-C),130.87(Ar-C),129.51(Ar-C),129.33(Ar-C),128.67(Ar-C),126.59(Ar-C),125.05(Ar-C),115.90(Ar-C),115.61(Ar-C),113.02(Ar-C),106.20(Ar-C),44.72(CH2),18.43(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 15 N2O5S + :455.0462;found:455.0466.

[0093] (Z)-2-(5-(3-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C9).Yellow solid;yield 43%;mp 267.8-268.2℃; 1 H NMR(500MHz,DMSO)δ10.88(s,1H,NH),8.00(s,1H,=CH),7.76(s,1H,Ar-H),7.73(s,1H,Ar-H),7.66(s,1H,Ar-H),7.60(s,3H,Ar-3H),7.48(d,J=10.0Hz,1H,Ar-H),6.28(s,1H,Ar-H),4.59(s,2H,CH2),2.40(s,3H,CH3); 13C NMR(126MHz,DMSO)δ167.05(C=O),165.32(C=O),164.86(C=O),160.21(C=O),153.90(CH),1 53.34(Ar-C),141.83(Ar-C),135.26(Ar-C),134.30(Ar-C),132.49(Ar-C),131.53(Ar-C),13 0.72(Ar-C),130.53(Ar-C),128.13(Ar-C),126.47(Ar-C),122.93(Ar-C),115.73(Ar-C),115 .44(Ar-C),112.85(Ar-C),106.02(Ar-C),44.53(CH2),18.26(CH3);HRMS(ESI-MS)m / z:[M+H] + calcdfor C 22 H 15 N2O5S + :455.0462;found:455.0467.

[0094] (Z)-2-(5-(4-chlorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C10).Yellow solid;yield 58%;mp 283.3-8℃. 1 H NMR(500MHz,DMSO)δ10.87(s,1H,NH),8.00(s,1H,,CH),7.73(d,J=10.0Hz,1H,Ar-H),7.69(s,1H,Ar-H),7.68(s,1H,Ar-H),7.6(s,1H,Ar-H), 1H,Ar-H),7.63(s,1H,Ar-H),7.62(s,1H,Ar-H),7.47(d,J=10.0Hz,1H,Ar-H),6.27(s,1H,Ar-H),4.59(s,2H,CH2),2.39(s,3H,3H) 13C NMR(126MHz,DMSO)δ167.14(C=O),165.42(C=O),164.87(C=O),160.21(C=O),153.89( =CH),153.31(Ar-C),141.83(Ar-C),135.76(Ar-C),132.76(Ar-C),132.14(Ar-2C),13 1.99(Ar-C),129.77(Ar-2C),126.43(Ar-C),121.86(Ar-C),115.71(Ar-C),115.43(Ar-C -C),112.84(Ar-C),106.02(Ar-C),44.50(CH2),18.26(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 15 N2O5S + :455.0462;found:455.0467.

[0095] (Z)-2-(5-(2-fluorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C11).Yellow solid;Yield 41;mp 291.3-291.6℃. 1 H NMR(500MHz,DMSO)δ10.89(s,1H,NH),7.97(s,1H,CH),7.74(d,J=10.0Hz,1H,Ar-H),7.67(s,1H,Ar-H),7.63(s,1H,Ar-H),7.63(s,1H,Ar-H),7.60(J). 0Hz,1H,Ar-H),7.48(d,J=10.0Hz,1H,Ar-H),7.43(s,1H,Ar-H),7.42(s,1H,Ar-H),6.28(s,1H,Ar-H),4.59(s,2H,CH2),2.40(s,3H,3H). 13 C NMR(126MHz,DMSO)δ167.30(C=O),165.45(C=O),165.04(C=O),161.0(d,J C-F =252.0Hz,=CH),160.02(Ar-C),153.81(d,J C-F =75.6Hz,Ar-C),142.02(Ar-C),133.71(d,J C-F=8.82Hz,Ar-C),129.60(Ar-C),126.65(Ar-C),126.01(d,J C-F =3.78Hz,Ar-C),125.43(d,J C-F =5.04Hz,Ar-C),124.15(Ar-C),121.18(d,J C-F =12.6Hz,Ar-C),116.91(Ar-C),116.74(Ar-C),115.93(Ar-C),115.64(Ar-C),1 13.05(Ar-C),106.22(Ar-C),44.75(CH2),18.46(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 15 FN2O5S + :439.0758;found:439.0759.

[0096] (Z)-2-(5-(3-fluorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C12).Yellow solid;yield 38%;mp 271.6-271.8℃. 1 H NMR(500MHz,DMSO)δ10.87(s,1H,NH),8.00(s,1H,,CH),7.72(d,J=10.0Hz,1H,Ar-H),7.65(s,1H,Ar-H),7.61(d,J5.0H,1H,7H),7.7H,Arz. J=5.0Hz,1H,Ar-H),7.48(d,J=10.0Hz,1H,Ar-H),7.45(s,1H,Ar-H),7.37(s,1H,Ar-H),6.26(s,1H,Ar-H),4.58(s,2H,CH2),2.38(CH)(CH) 13C NMR(126MHz,DMSO)δ167.28(C=O),165.53(C=O),165.02(C=O),163.69(C=O),161.74(=CH),160.36(Ar-C),154.05(Ar-C),153.46(Ar-C),141.99(Ar-C),135.59(d,J=7.7Hz,Ar-C),132.83(Ar-C),131.97(Ar-C),131.91(Ar-C),126.59(Ar-C),126.14(d,J=2.5Hz,Ar-C),122.97(Ar-C),118.07(d,J C-F =21.4Hz,Ar-C),117.46(d,J C-F =22.7Hz,Ar-C),115.73(d,J C-F =35.3Hz,Ar-C),109.59(d,J C-F =859.3Hz,Ar-C),44.68(CH2),18.42(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 15 FN2O5S + :439.0758;found:439.0761.

[0097] (Z)-2-(5-(4-fluorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C13).Yellow solid;yield 53%;mp 286.9-287.3℃; 1 H NMR(500MHz,DMSO)δ10.87(s,1H,NH),8.03(s,1H,=CH),7.76(d,J=5.0Hz,1H,Ar-H),7.75(s,1H,Ar-H),7.74(s,1H,Ar-H),7.67(s,1H,Ar-H),7.48(d,J=10.0Hz,1H,Ar-H),7.43(d,J=10.0Hz,1H,Ar-H),7.40(s,1H,Ar-H),6.29(s,1H,Ar-H),4.59(s,2H,CH2),2.40(s,3H,CH3); 13C NMR(126MHz,DMSO)δ167.26(C=O),165.47(C=O),164.89(C=O),162.38(C=O),160.19(CH) ,153.88(Ar-C),153.29(Ar-C),141.83(Ar-C),133.06(Ar-C),132.99(Ar-C),129.77(d,J C-F =3.8Hz,Ar-C),126.41(Ar-C),120.80(d,J C-F =2.5Hz,Ar-C),116.89(d,J C-F =21.4Hz,Ar-C),115.69(Ar-C),115.41(Ar-C),112.82(Ar-C),106.00(Ar-C),44.47(CH2),18.24(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd forC 22 H 15 FN2O5S + :439.0758;found:439.0763.

[0098] (Z)-2-(5-(2-bromobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C14). 1 H NMR(500MHz,DMSO)δ10.88(s,1H,NH),8.05(s,1H,=CH),7.84(d,J=10.0Hz ,1H,Ar-H),7.75(d,J=10.0Hz,1H,Ar-H),7.67(s,1H,Ar-H),7.63(d,J=5. 0Hz,1H,Ar-H),7.60(d,J=10.0Hz,1H,Ar-H),7.49(s,1H,Ar-H),7.46(d,J =10.0Hz,1H,Ar-H),6.28(s,1H,Ar-H),4.59(s,2H,CH2),2.40(s,3H,CH3); 13C NMR(126MHz,DMSO)δ167.00(C=O),164.92(C=O),164.66(C=O),160.03(C=O),153.72(CH),1 53.15(Ar-C),141.64(Ar-C),133.79(Ar-C),132.69(Ar-C),132.49(Ar-C),131.69(Ar-C),12 9.30(Ar-C),128.85(Ar-C),126.28(Ar-C),125.42(Ar-C),124.75(Ar-C),115.56(Ar-C),115 .27(Ar-C),112.69(Ar-C),105.86(Ar-C),44.38(CH2),18.09(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 15 BrN2O5S + :498.9957;found:498.9965.

[0099] (Z)-2-(5-(3-bromobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C15). 1 H NMR(500MHz,DMSO)δ10.86(s,1H,NH),7.99(s,1H,,CH),7.88(s,1H,Ar-H ),7.73(d,J=10.0Hz,1H,Ar-H),7.71(d,J=5.0Hz,1H,Ar-H),7.65(s,1H,A r-H),7.64(d,J=5.0Hz,1H,Ar-H),7.52(t,J=5.0Hz,1H,Ar-H),7.47(d,J= 15.0Hz,1H,Ar-H),6.27(s,1H,Ar-H),4.58(s,2H,CH2),2.39(s,3H,CH3); 13C NMR(126MHz,DMSO)δ167.03(C=O),165.29(C=O),164.84(C=O),160.19(C=O),153.88(CH),1 53.30(Ar-C),141.81(Ar-C),135.49(Ar-C),133.58(Ar-C),133.40(Ar-C),132.43(Ar-C),13 1.70(Ar-C),128.44(Ar-C),126.44(Ar-C),122.86(Ar-C),122.80(Ar-C),115.71(Ar-C),115 .42(Ar-C),112.84(Ar-C),106.00(Ar-C),44.52(CH2),18.26(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 15 BrN2O5S + :498.9957;found:498.9965.

[0100] (Z)-2-(5-(4-bromobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C16).Yellow sold;yield 62;mp 294.6-9℃. 1 H NMR(500MHz,DMSO)δ10.85(s,1H,NH),7.96(s,1H,CH),7.75(s,1H,Ar-H),7.74(s,1H,Ar-H),7.71(d,J=5.0Hz,1H,Ar-H),7.64(s,1H,Ar-H). H,Ar-H),7.60(s,1H,Ar-H),7.58(s,1H,Ar-H),7.45(d,J=15.0Hz,1H,Ar-H),6.26(s,1H,Ar-H),4.57(s,2H,CH2),2.38(s,3H,CH3) 13C NMR(126MHz,DMSO)δ167.11(C=O),165.41(C=O),164.85(Ar-C),160.18(Ar-C),153.87 (Ar-C),153.28(Ar-C),141.81(Ar-C),132.85(Ar-C),132.69(Ar-2C),132.30(Ar-C),1 32.26(Ar-2C),126.41(Ar-C),124.71(Ar-C),121.93(Ar-C),115.69(Ar-C),115.41(A r-C),112.83(Ar-C),106.00(Ar-C),44.50(CH2),18.25(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 15 BrN2O5S + :498.9957;found:498.9965.

[0101] (Z)-2-(5-(2-hydroxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C17). 1 H NMR(500MHz,DMSO)δ10.86(s,1H,NH),10.65(s,1H,OH),8.18(s,1H,CH), 7.74(d,J=5.0Hz,1H,Ar-H),7.66(s,1H,Ar-H),7.48(d,J=10.0Hz,1H,Ar-H H),7.40(d,J=10.0Hz,1H,Ar-H),7.35(t,J=10.0Hz,1H,Ar-H),6.98(t,J= 10.0Hz,2H,Ar-2H),6.28(s,1H,Ar-H),4.57(s,2H,CH2),2.39(s,3H,CH3); 13C NMR(126MHz,DMSO)δ167.71(C=O),165.75(C=O),165.00(C=O),160.20(C=O),157.70(CH),1 53.89(Ar-C),153.30(Ar-C),141.86(Ar-C),133.04(Ar-C),129.37(Ar-C),128.97(Ar-C),12 6.42(Ar-C),120.09(Ar-C),120.00(Ar-C),119.60(Ar-C),116.52(Ar-C),115.69(Ar-C),115 .41(Ar-C),112.82(Ar-C),105.99(Ar-C),44.37(CH2),18.25(CH3);HRMS(ESI-MS)m / z:[M+H] + calcdfor C 22 H 16 N2O6S + :437.0801;found:437.0807.

[0102] (Z)-2-(5-(3-hydroxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C18). 1 H NMR(500MHz,DMSO)δ10.88(s,1H,NH),9.94(s,1H,OH),7.90(s,1H,,CH),7.75(d, J=10.0Hz,1H,Ar-H),7.66(s,1H,Ar-H),7.48(d,J=10.0Hz,1H,Ar-H),7.36(t,J) 10.0Hz,1H,Ar-H),7.10(d,J=10.0Hz,1H,Ar-H),7.04(s,1H,Ar-H),6.92(d,J=10 .0Hz,1H,Ar-H),6.28(s,1H,Ar-H),4.57(s,2H,CH2),2.40(d,J=10.0Hz,3H,CH3); 13C NMR(126MHz,DMSO)δ167.34(C=O),165.42(C=O),164.84(C=O),160.14(C=O),158.09(=CH),153.78(Ar-C),153.27(Ar-C),141.71(Ar-C),134.17(Ar-C),134.15(Ar-C),130.72(Ar-C),126.37(Ar-C),121.68(Ar-C),120.79(Ar-C),118.36(Ar-C),116.25(Ar-C),115.63(Ar-C),115.35(Ar-C),112.73(Ar-C),105.91(Ar-C),44.31(CH2),18.16(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 16 N2O6S + :437.0801;found:437.0809.

[0103] (Z)-2-(5-(4-hydroxybenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C19).Yellow sold;yield 32%;mp 328.8-329.2℃; 1 H NMR(500MHz,DMSO)δ10.85(s,1H,NH),10.43(s,1H,OH),7.89(s,1H,=CH),7.73(d,J=10.0Hz,1H,Ar-H),7.66(s,1H,Ar-H),7.54(s,1H,Ar-H),7.52(s,1H,Ar-H),7.47(d,J=10.0Hz,1H,Ar-H),6.95(s,1H,Ar-H),6.93(s,1H,Ar-H),6.27(s,1H,Ar-H),4.56(s,2H,CH2),2.39(s,3H,CH3); 13C NMR(126MHz,DMSO)δ167.49(C=O),165.63(C=O),164.95(C=O),160.52(C=O),160.12( =CH),153.80(Ar-C),153.23(Ar-C),141.78(Ar-C),134.40(Ar-C),132.95(Ar-C),12 6.34(Ar-C),123.93(Ar-C),116.64(Ar-C),116.38(Ar-C),115.59(Ar-C),115.33(Ar-C -C),112.73(Ar-C),105.90(Ar-C),44.26(CH2),18.17(CH3);HRMS(ESI-MS)m / z:[M+H] + calcd for C 22 H 16 N2O6S + :437.0801;found:437.0811.

[0104] (Z)-2-(5-(3,4-difluorobenzylidene)-2,4-dioxothiazolidin-3-yl)-N-(4-methyl-2-oxo-2H-chromen-7-yl)acetamide(C20).Yellow solid;yield 32%;mp 269.8-37ℛ. 1 H NMR(500MHz,DMSO)δ10.87(s,1H,NH),7.99(s,1H,CH),7.80(d,J=10.0Hz,1H,Ar-H),7.74(d,J=5.0Hz,1H,Ar-H),7.66(s,1H,A). r-H),7.63(s,1H,Ar-H),7.53(s,1H,Ar-H),7.47(d,J=10.0Hz,1H,Ar-H),6.28(s,1H,Ar-H),4.59(s,2H,CH2),2.39(s,3H,CH3)! 13 C NMR(126MHz,DMSO)δ166.60(CO),164.88(CO),164.39(CO),159.75(CO),153.45(CH),152.86(Ar-C),150.95(d,JJ). C-F =110.9Hz,Ar-C),148.91(d,J C-F=90.7Hz,Ar-C),141.37(Ar-C),131.44(Ar-C),130.42(Ar-C),126.91(Ar-C),126.86(Ar-C),125.99(Ar-C),122.02(Ar-C),118.98(d,J C-F =119.0Hz, Ar-C), 115.27 (Ar-C), 114.98 (Ar-C), 112.40 (Ar-C), 105.56 (Ar-C), 44.08 (CH2), 17.81 (CH3).; HRMS (ESI-MS) m / z: [M+H] + calcdfor C 22 H 14 F2N2O5S + :457.0668; found:457.0670.

[0105] Example 2

[0106] This embodiment provides an α-glucosidase inhibitory activity test for coumarin derivatives containing a thiazolidinedione structure.

[0107] 1. Preparation of reagents and standard solutions

[0108] (1) 100mM phosphate buffer (PBS, pH=6.8): Weigh a certain amount of potassium dihydrogen phosphate and disodium hydrogen phosphate, dissolve them in ultrapure water, and use them to dissolve and dilute the reagent.

[0109] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100mM PBS to the enzyme with an activity of 100U to prepare a working concentration of 0.05U / mL, and then dispense and freeze.

[0110] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoside (PNPG), add 100mM PBS solution to dissolve, prepare a substrate working solution with a concentration of 0.25mM, vortex mix well, and prepare fresh before each experiment.

[0111] Preparation of test drug: Accurately weigh an appropriate amount of the test drug, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20℃ protected from light. Before the experiment, dilute with DMSO to different required concentrations (0–200 μM), with the DMSO content equal to 5%.

[0112] 2. Experimental Procedure

[0113] (1) Add 10 μL of α-glucosidase with a working concentration of 0.05 U / mL, 130 μL of phosphate buffer with a concentration of 100 mM (pH = 6.8), and 10 μL of compounds of different concentrations (coumarin derivatives C1 to C20 with a thiazolidinedione structure prepared in Example 1) to a 96-well plate in sequence. Replace 10 μL of the compound with 10 μL of DMSO with a content of 5% in the blank control group. Use acarbose as a positive control. Set up 4 replicates for each group. Incubate the enzyme reaction system on a microplate reader at 37°C for 10 min.

[0114] (2) Subsequently, 50 μL of substrate PNPG was added to the enzyme reaction system to start the enzyme reaction. The microplate was placed on the microplate reader and incubated at 37°C for 15 min. During the incubation, the time was evenly distributed 3 times, and the reading was taken once at a wavelength of 405 nm for each time period. The readings were recorded as OD1, OD2, and OD3.

[0115] (3) The α-glucosidase inhibitory activity of the test compound is calculated according to the following formula:

[0116] Inhibition rate (%) = [(OD3-OD)-(OD1-OD)] / OD3-OD × 100%

[0117] Where OD represents the absorbance value of the blank control group. Data processing: Data were analyzed and processed using MS Excel, and the half-maximal inhibitory concentration (IC50) was calculated using Origin 9.1. 50 IC 50 This represents the concentration of the test compound required to inhibit α-glucosidase activity by 50% under the experimental conditions stated.

[0118] 3. Results Analysis

[0119] The α-glucosidase inhibitory activity of the synthesized compounds was evaluated using in vitro enzymatic experiments, and the results are shown in Table 2.

[0120] Table 2 Evaluation of the in vitro inhibitory activity of compounds e1–e36 against α-glucosidase

[0121]

[0122] The positive control drug, acarbose, had an IC50 value of [missing information]. 50 The concentration was 655.01 μM. Three derivatives (C15, C3, and C16) with good α-glucosidase inhibitory activity were screened. The inhibition rates of C15, C3, and C16 were 5.12 ± 0.98 μM, 6.58 ± 0.75 μM, and 7.33 ± 1.26 μM, respectively, which were 34–128 times that of acarbose. Among them, C15 showed the best inhibition rate, and its half-maximal inhibitory concentration (IC50) is shown in the figure below. Figure 1 As shown in the figure, these compounds exhibit strong binding affinity when interacting with α-glucosidase. Therefore, compound C15 can serve as a lead compound for developing α-glucosidase inhibitors with anti-diabetic activity.

[0123] Example 3

[0124] Enzyme kinetics test

[0125] The α-glucosidase inhibitory activity of the synthesized active compound was evaluated using in vitro enzyme kinetic experiments.

[0126] 1. Preparation of reagents and standard solutions

[0127] 1) 100mM phosphate buffer (PBS, pH 6.8): Weigh a certain amount of potassium dihydrogen phosphate and disodium hydrogen phosphate, dissolve them in ultrapure water, and use them to dissolve and dilute the reagent.

[0128] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100mM PBS to the enzyme with an activity of 100U to prepare working concentrations of 0.0375U / mL, 0.05U / mL, 0.0625U / mL and 0.075U / mL, and then dispense and freeze.

[0129] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoside (PNPG), add 100mM PBS solution to dissolve, prepare a substrate working solution with a concentration of 0.25mM, vortex mix well, and prepare fresh before each experiment.

[0130] Preparation of test drug: Accurately weigh an appropriate amount of the test drug, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20℃ protected from light. Before the experiment, dilute with DMSO to different required concentrations (0–200 μM), with the DMSO content equal to 5%.

[0131] 2. Experimental Procedure

[0132] (1) Add 10 μL of α-glucosidase with concentrations of 0.0375 U / mL, 0.05 U / mL, 0.0625 U / mL and 0.075 U / mL, respectively, 130 μL of 100 mM phosphate buffer (pH 6.8), and 10 μL of compound of different concentrations (coumarin derivative C15 with thiazolidinedione structure prepared in Example 1) to a 96-well plate. Replace 10 μL of compound with 10 μL of DMSO with 5% DMSO for the blank control group. Use acarbose as a positive control. Set up 4 replicates for each group. Incubate the enzyme reaction system on a microplate reader at 37°C for 10 min.

[0133] (2) Subsequently, 50 μL of substrate PNPG with a concentration of 0.25 mM was added to the enzyme reaction system to start the enzyme reaction. The microplate was placed on the microplate reader and incubated at 37°C for 15 min. During the incubation process, the time was evenly divided into 3 times, and the reading was taken once at a wavelength of 405 nm for each time period. The readings were recorded as OD1, OD2, and OD3.

[0134] (3) Data processing: MS Excel was used to analyze and process the data. The reaction rate of the enzyme reaction system was ΔOD / min.

[0135] 3. Results Analysis

[0136] The results of the enzyme kinetic inhibition type evaluation experiment are as follows: Figure 2 As shown. By Figure 2 It can be seen that the inhibitor is a reversible inhibitory effect.

[0137] Example 4

[0138] Substrate dynamics experiment

[0139] The α-glucosidase inhibitory activity of the synthesized active compounds was evaluated using in vitro substrate kinetic experiments.

[0140] 1. Preparation of reagents and standard solutions

[0141] (1) 100mM phosphate buffer (PBS, pH 6.8): Weigh a certain amount of potassium dihydrogen phosphate and disodium hydrogen phosphate, dissolve them in ultrapure water, and use them to dissolve and dilute the reagent.

[0142] (2) Preparation of α-glucosidase solution: Add an appropriate amount of 100mM PBS to the enzyme with an activity of 100U to prepare a working concentration of 0.05U / mL, and then dispense and freeze.

[0143] (3) Substrate preparation: Accurately weigh an appropriate amount of 4-nitrophenyl-D-pyranoside (PNPG), add 100mM PBS solution to dissolve, and prepare substrate working solutions with concentrations of (0.25mM, 0.5mM, 0.75mM, 1mM). Vortex mix well and prepare fresh before each experiment.

[0144] Preparation of test drug: Accurately weigh an appropriate amount of the test drug, dissolve it in DMSO to prepare a 10 mM stock solution, and store it at -20℃ protected from light. Before the experiment, dilute with DMSO to different required concentrations (0–200 μM), with the DMSO content equal to 5%.

[0145] 2. Experimental Procedure

[0146] (1) Add 10 μL of 0.5 U / mL α-glucosidase, 130 μL of 100 mM phosphate buffer (pH 6.8), and 10 μL of different concentrations of compound (coumarin derivative C15 with thiazolidinedione structure prepared in Example 1) to a 96-well plate in sequence. Replace 10 μL of compound with 10 μL of 5% DMSO for the blank control group. Use acarbose as a positive control. Set up 4 replicates for each group. Incubate the enzyme reaction system at 37°C for 10 min on a microplate reader.

[0147] (2) Subsequently, 50 μL of substrate PNPG of different concentrations was added to the enzyme reaction system to start the enzyme reaction. The microplate was placed on the microplate reader and incubated at 37°C for 15 min. During the incubation, the time was evenly distributed 3 times, and the reading was taken once at a wavelength of 405 nm for each time period. The readings were recorded as OD1, OD2, and OD3.

[0148] (3) Data processing: MS Excel was used to analyze and process the data. The reaction rate of the enzyme reaction system was ΔOD / min.

[0149] 3. Results Analysis

[0150] The experimental results for evaluating the type of substrate kinetic inhibition are as follows: Figure 3 As shown, Figure 3 The substrate kinetics of the coumarin derivative C15 containing the thiazolidinedione structure on α-glucosidase in vitro. Figure 3 As can be seen, all the straight lines almost intersect at the x-axis, and the Michaelis-Menten constant (Km) remains unchanged, proving that compound C15 has a non-competitive inhibitory effect.

[0151] Example 5

[0152] Cytotoxicity test

[0153] Cell culture was carried out in an incubator (37°C, containing 5% CO2) using DMEM culture medium.

[0154] Human kidney epithelial cell line (293 cells) and normal human hepatocytes (LO2 cells) were seeded into 96-well plates and cultured for 24 h. C15 (1–64 μM) was added, and the cells were cultured continuously for 48 h. The culture medium was then removed, and 100 μL of fresh culture medium and 10 μL of MTT solution (5 mg / mL) were added. The cells were incubated for 4 h, the culture medium was discarded, and 100 μL of formazan dissolved in DMSO was added. After incubation for 15 minutes, the absorbance at 490 nm was measured, and the viability was calculated.

[0155] 2. Results Analysis

[0156] The toxicity of compound C15 to 293 and LO2 cells was examined at cell-administered concentrations (1–64 μM). Results are as follows: Figure 4 and Figure 5 As shown, at concentrations of 1–64 μM, compound C15 has virtually no effect on cell viability, indicating that C15 has good safety at low concentrations.

[0157] In summary, this invention provides a series of coumarin derivatives containing thiazolidinedione structures and determines their inhibitory activity against α-glucosidase. The results show that most of the synthesized derivatives exhibit significant inhibitory activity against α-glucosidase, particularly compounds C15, C3, and C16, with inhibition rates of 5.12±0.98 μM, 6.58±0.75 μM, and 7.33±1.26 μM, respectively, which are 34–128 times higher than that of acarbose. C15 showed the best inhibition rate, as shown in the half-maximal inhibitory concentration (IC50) plot. Figure 1 As shown in the figure, these compounds exhibit strong binding affinity when interacting with α-glucosidase. Therefore, compound C15 can serve as a lead compound for developing α-glucosidase inhibitors with anti-diabetic activity.

[0158] The embodiments of the present invention have been described in detail above. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. A coumarin derivative containing a thiazolidinedione structure, characterized in that, The coumarin derivatives have the following structural formula: In the formula, R is selected from substituted or unsubstituted phenyl; the substitution is monosubstituted or polysubstituted; the substituent is selected from at least one of halogen, methyl, methoxy, and hydroxyl.

2. The coumarin derivative containing a thiazolidinedione structure according to claim 1, characterized in that, In the coumarin derivatives, R is selected from one of the following groups:

3. A method for preparing a coumarin derivative containing a thiazolidinedione structure as described in any one of claims 1 to 2, the method comprising the following steps: The coumarin derivative containing the thiazolidinedione structure was obtained by reacting compound 2 and compound 5 containing the R group as raw materials; The structures of compounds 2 and 5 are as follows:

4. The preparation method according to claim 3, characterized in that, The molar ratio of compound 2 to compound 5 containing the R group is (1.2 to 1.5):

1.

5. The preparation method according to claim 3, characterized in that, The reaction temperature is 20–30°C.

6. The preparation method according to claim 3, characterized in that, The reaction time is 1 to 3 days.

7. The preparation method according to claim 3, characterized in that, The stirring speed for the reaction is 400–600 rpm.

8. The preparation method according to claim 3, characterized in that, The reaction process also includes a washing and filtration step.

9. An α-glucosidase inhibitor, characterized in that, Including coumarin derivatives containing a thiazolidinedione structure as described in any one of claims 1 to 2.

10. The use of a coumarin derivative containing a thiazolidinedione structure as described in any one of claims 1 to 2 in the preparation of a medicament for the prevention and / or treatment of diabetes.

11. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises a coumarin derivative containing a thiazolidinedione structure as described in any one of claims 1 to 2 and / or a pharmaceutically acceptable salt thereof.

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