Coumarin derivative containing thiazolidinedione structure as well as preparation method and application of coumarin derivative

Abstract

The invention discloses a coumarin derivative containing a thiazolidinedione structure as well as a preparation method and application of the coumarin derivative, and belongs to the field of medicinal chemistry. The coumarin derivative containing the thiazolidinedione structure has a structure as shown in the following formula: # imgabs0 #, in the formula, R represents substituted or unsubstituted phenyl, and the substitution is optional substitution. A series of coumarin derivatives containing thiazolidinedione structures are designed and synthesized, the coumarin derivatives containing the thiazolidinedione structures show high alpha-glucosidase inhibitory activity, the minimum IC50 value of the coumarin derivatives containing the thiazolidinedione structures is 5.12 + / -0.98 mu M and is 34-128 times that of acarbose, and the coumarin derivatives containing the thiazolidinedione structures can be used for treating and / or preventing diabetes mellitus. The invention also provides a preparation method and application of the coumarin derivative containing the thiazolidinedione structure.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical chemistry, and in particular to a coumarin derivative containing a thiazolidinedione structure, and a preparation method and application thereof. Background Art

[0002] At present, diabetes has become a global problem, seriously endangering human health. Although people's health awareness has improved, the incidence of diabetes is still rising. If blood sugar levels are not effectively controlled in time, it will induce a series of functional disorders and damages in the human body, seriously threatening the health and safety of patients. Diabetes is divided into type Ⅰ diabetes and type Ⅱ diabetes. Among them, type Ⅰ diabetes, also known as insulin-dependent diabetes mellitus (IDDM), accounts for about 10% of diabetic patients. It is mainly caused by severe damage to the pancreatic β cells that secrete insulin in the body, resulting in insufficient endogenous insulin secretion. It has a certain hereditary nature. Type Ⅱ 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, eating habits, etc., accounting for more than 90% of the number of patients.

[0003] There are many oral hypoglycemic drugs on the market, which act on different targets through different pathways. Sulfonylurea drugs (glibenclamide, glimepiride and mesitylamide) are insulin secretagogues that can activate voltage-gated Ca 2+Channels to stimulate pancreatic beta cells to release insulin. Biguanides, such as metformin, are considered first-line treatments for diabetes. Their mechanism of action is to lower blood sugar by inhibiting hepatic gluconeogenesis and hepatic bile sugar output. Sodium-glucose transporter inhibitors (SGLT2) drugs, represented by dapagliflozin, can inhibit glucose reabsorption, thereby controlling blood sugar and reducing calories. In addition, another way to maintain blood sugar levels is to inhibit the hydrolysis of α-glucosidase, thereby reducing postprandial blood sugar. α-Glucosidase helps 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 the digestion and absorption of carbohydrates, thereby reducing insulin demand. In addition, α-glucosidase inhibitors can lead to a long-term and sustained increase in endogenous GLP-1, thereby stimulating insulin secretion and reducing glucagon secretion, thereby achieving the effect of controlling postprandial hyperglycemia. Currently available α-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, it is very meaningful to develop drugs with good efficacy and low side effects. In recent years, many new studies on α-glucosidase inhibitors have been carried out, which may help to discover new α-glucosidase inhibitors in the future.

[0004] Coumarin is a naturally occurring organic compound that has become a hot topic of research due to its wide range of biological activities, such as antioxidant, anti-inflammatory, antibacterial, and anticancer. However, there are few studies on coumarin compounds as α-glucosidase inhibitors. Therefore, combining the coumarin skeleton with other biologically active structures may produce new and effective α-glucosidase inhibitors. Thiazolidinediones are compounds with a wide range of biological activities and have been shown to have significant effects in the fields of antibacterial, anti-tuberculosis, and anti-cancer. According to existing literature, 5-benzylidenethiazolidine-2,4-dione IC 50 =98.88±1.11μM, coumarin analog 3-acetylcoumarin IC 50 =1.5×10 5 μM.

[0005] At present, no coumarin derivatives containing thiazolidinedione structure have been disclosed, so it is still of great significance to develop some new derivatives with α-glucosidase inhibitory effect. Summary of the invention

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

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

[0008] The present invention also provides the application of the above coumarin derivatives containing thiazolidinedione structure.

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

[0010]

[0011] In the formula, R is selected from substituted or unsubstituted phenyl, 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] The present invention uses the coumarin structure as the parent core to design and synthesize a series of coumarin derivatives containing thiazolidinedione structure, among which the coumarin derivative C15 substituted with Br at the meta-position exhibits a strong α-glucosidase inhibitory effect, and its IC 50 The maximum value is 5.12±0.98μM, which is 128 times that of acarbose. Compared with a single thiazolidinedione structural derivative and a single coumarin derivative, the compound of the present invention has a significant α-glucosidase inhibitory effect. Kinetic studies have shown that C15 is a reversible, non-competitive inhibitor and can be used as an α-glucosidase inhibitor to treat or prevent diabetes.

[0014] According to some embodiments of the present invention, the substitution is mono-substitution or poly-substitution.

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

[0016] According to some embodiments of the present invention, R is selected from the group shown below:

[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] Compound 2 and compound 5 containing an R group are used as raw materials to react to obtain the coumarin derivative containing a thiazolidinedione structure;

[0020] Among them, the structures of compound 2 and compound 5 are as follows:

[0021]

[0022] According to some embodiments of the present invention, the molar ratio of compound 2 to compound 5 is (1.2-1.5): 1. For example, it may 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 of 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, about 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, about 1 day, 2 days, or 3 days.

[0026] According to some embodiments of the present invention, the reaction stirring speed is about 400-600 rpm, for example, about 400 rpm, 500 rpm, or 600 rpm.

[0027] According to some embodiments of the present invention, the post-reaction treatment step includes: adding an appropriate amount of water to the reaction system, and washing and filtering the solid produced with water.

[0028] According to some embodiments of the present invention, the method for preparing compound 2 comprises the following steps:

[0029] 7-Amino-4-methylcoumarin (Compound 1) is used as a starting material and reacts with bromoacetyl bromide in the presence of a base at room temperature to obtain Compound 2.

[0030] Wherein, 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 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:(1.2-1.4), for example, 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 environment.

[0037] According to some embodiments of the invention, the inert atmosphere comprises 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 comprises the following steps:

[0040] At room temperature, 2,4-thiazolidinedione (compound 4) is used as a starting material and reacts with benzaldehyde substituted with an R group in the presence of a base to obtain compound 5.

[0041] Among them, 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 the benzaldehyde substituted with an R group is (1-1.2): 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 of the reaction is anhydrous ethanol (EtOH).

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

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

[0047] According to the third aspect of the present invention, an α-glucosidase inhibitor is provided, which includes the above-mentioned coumarin derivative containing a thiazolidinedione structure.

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

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

[0050] According to a fifth aspect of the present invention, a pharmaceutical composition is provided, comprising the above-mentioned coumarin derivative containing a thiazolidinedione structure and / or a pharmaceutically acceptable salt thereof.

[0051] According to some embodiments of the present invention, the dosage form of the drug is tablet, capsule, oral solution or injection.

[0052] The structural formulas in the present invention appear Indicates the attachment site of a group.

[0053] The term "pharmaceutically acceptable" as used herein refers to substances that are acceptable for pharmaceutical use from a toxicological point of view and do not adversely interact with the active ingredients.

[0054] Other features and advantages of the present invention will be set forth in the description which follows, and in part will be apparent from the description, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0056] Figure 1 A graph of the half-maximal inhibition concentration of α-glucosidase in vitro by providing a coumarin derivative C15 containing a thiazolidinedione structure as an α-glucosidase inhibitor for Example 2 of the present invention;

[0057] Figure 2 This is a diagram showing the enzyme kinetics of the thiazolidinedione structure-containing coumarin derivative C15 in Example 3 of the present invention as an α-glucosidase inhibitor on α-glucosidase in vitro;

[0058] Figure 3 This is a kinetic diagram of the substrate of α-glucosidase in vitro by using the coumarin derivative C15 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 results of a cytotoxicity test on LO2 cells of the coumarin derivative C15 containing a thiazolidinedione structure as an α-glucosidase inhibitor in Example 5 of the present invention;

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

[0061] The following will be combined with the embodiments to clearly and completely describe the concept of the present invention and the technical effects produced, so as to fully understand the purpose, characteristics and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present invention.

[0062] In the description of the present invention, if there is a description of first, second, etc., it is only for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0063] When a numerical range is disclosed herein, the above range is considered to be continuous and includes the minimum and maximum values ​​of the range, as well as every value between such minimum and maximum values. Further, when a range refers to an integer, every integer between the minimum and maximum values ​​of the range is included. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be combined. In other words, unless otherwise indicated, all ranges disclosed herein should be understood to include any and all subranges included therein.

[0064] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of the present invention.

[0065] Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

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

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

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

[0069] Triethylamine CAS number: 121-44-8;

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

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

[0072] Example 1

[0073] This example prepares a series of coumarin derivatives containing thiazolidinedione structure, and the preparation route is as follows:

[0074]

[0075] The preparation method comprises 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 completed, the solid was collected and then washed with 100 ml of water to obtain compound (2).

[0077] 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 and filtered, washed with n-hexane (50 mL), and the solid was collected to obtain compound (5).

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

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

[0080] Table 1 Structural formula of R group

[0081]

[0082]

[0083]

[0084] The structures of C1 to C20 were characterized by NMR, MS and melting point. The properties, yields, NMR and mass spectrometry results of each compound are shown below:

[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 example provides a test for the α-glucosidase inhibitory activity of coumarin derivatives containing a thiazolidinedione structure.

[0107] 1. Preparation of reagents and standard solutions

[0108] (1) 100 mM 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 100 mM PBS to an enzyme with an activity of 100 U to a working concentration of 0.05 U / mL, and freeze in aliquots.

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

[0111] Preparation of test drugs: Accurately weigh the appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10mM stock solution, and store it at -20°C in the dark. Dilute it with DMSO to different required concentrations (0-200μM) before the experiment, and the DMSO content is equal to 5%.

[0112] 2. Experimental steps

[0113] (1) 10 μL of α-glucosidase with a working concentration of 0.05 U / mL, 130 μL of phosphate buffer (pH = 6.8) with a concentration of 100 mM, and 10 μL of compounds with different concentrations (coumarin derivatives C1 to C20 containing thiazolidinedione structure prepared in Example 1) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of DMSO with a content equal to 5%, and acarbose was used as a positive control. Four replicate wells were set in parallel for each group, and the enzyme reaction system was placed on a microplate reader and incubated at 37° C. for 10 min.

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

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

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

[0117] OD represents the absorbance value of the blank control group. Data processing: MS Excel was used to analyze and process the data, and Origin 9.1 was used to calculate the half inhibitory concentration (IC 50 ), IC 50 It represents the concentration of the test compound required to inhibit the activity of α-glucosidase by 50% under the experimental conditions.

[0118] 3. Results Analysis

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

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

[0121]

[0122] The IC value of the positive control drug acarbose 50 The inhibitory rates of C15, C3 and C16 were 5.12±0.98μM, 6.58±0.75μM and 7.33±1.26μM, respectively, which were 34 to 128 times that of acarbose. Among them, C15 had the best inhibitory rate, and its half inhibition concentration was shown in the figure below. Figure 1 The results showed that these compounds showed strong binding affinity when interacting with α-glucosidase. Therefore, compound C15 can be used as a lead compound for the development of α-glucosidase inhibitors with anti-diabetic activity.

[0123] Example 3

[0124] Enzyme kinetics assay

[0125] The in vitro enzyme kinetics experiment was used to evaluate the α-glucosidase inhibitory activity of the synthesized active compounds:

[0126] 1. Preparation of reagents and standard solutions

[0127] 1) 100 mM 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 reagents.

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

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

[0130] Preparation of test drugs: Accurately weigh the appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10mM stock solution, and store it at -20°C in the dark. Dilute it with DMSO to different required concentrations (0-200μM) before the experiment, and the DMSO content is equal to 5%.

[0131] 2. Experimental steps

[0132] (1) 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, 130 μL of 100 mM phosphate buffer (pH 6.8), and 10 μL of compounds with different concentrations (the coumarin derivative C15 containing a thiazolidinedione structure prepared in Example 1) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of DMSO with a content equal to 5%, and acarbose was used as a positive control. Four replicate wells were set in parallel for each group, and the enzyme reaction system was placed on a microplate reader and incubated at 37° C. for 10 min.

[0133] (2) Subsequently, 50 μL of 0.25 mM PNPG was added to the enzyme reaction system to initiate the enzyme reaction. The microplate was placed on a microplate reader and incubated at 37°C for 15 min. During the incubation process, the incubation time was evenly distributed three times. The readings were taken once at a wavelength of 405 nm during 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 △OD / min.

[0135] 3. Results Analysis

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

[0137] Example 4

[0138] Substrate kinetics experiments

[0139] The in vitro substrate kinetics experiment was used to evaluate the α-glucosidase inhibitory activity of the synthesized active compounds:

[0140] 1. Preparation of reagents and standard solutions

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

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

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

[0144] Preparation of test drugs: Accurately weigh the appropriate amount of the drug to be tested, dissolve it in DMSO to prepare a 10mM stock solution, and store it at -20°C in the dark. Dilute it with DMSO to different required concentrations (0-200μM) before the experiment, and the DMSO content is equal to 5%.

[0145] 2. Experimental steps

[0146] (1) 10 μL of 0.5 U / mL α-glucosidase, 130 μL (pH 6.8) of 100 mM phosphate buffer, and 10 μL of compounds of different concentrations (the coumarin derivative C15 containing a thiazolidinedione structure prepared in Example 1) were added to a 96-well plate in sequence. The blank control group was replaced with 10 μL of 5% DMSO to replace 10 μL of the compound. Acarbose was used as a positive control. Four replicate wells were set in parallel for each group. The enzyme reaction system was placed on a microplate reader and incubated at 37° C. for 10 min.

[0147] (2) Subsequently, 50 μL of substrate PNPG of different concentrations was added to the enzyme reaction system to initiate the enzyme reaction. The microplate was placed on a microplate reader and incubated at 37°C for 15 min. During the incubation process, the readings were evenly distributed three times and read once at a wavelength of 405 nm during 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 △OD / min.

[0149] 3. Results Analysis

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

[0151] Example 5

[0152] Cytotoxicity assay

[0153] The DMEM medium was placed in an incubator (37°C, containing 5% CO2) for cell culture.

[0154] Human renal epithelial cell line (293 cells) and human normal liver cells (LO2 cells) were inoculated in 96-well plates, cultured for 24 hours, C15 (1-64 μM) was added, and cultured for 48 hours. Then the culture medium was removed, 100 μL of fresh culture medium and 10 μL of MTT solution (5 mg / mL) were added, and incubated for 4 hours. 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 survival rate was calculated.

[0155] 2. Results Analysis

[0156] The toxicity of compound C15 to 293 cells and LO2 cells at the cell administration concentration (1-64 μM) was tested. Figure 4 and Figure 5 As shown, when the concentration is 1-64 μM, compound C15 has little effect on cell viability, indicating that C15 has good safety at low concentrations.

[0157] In summary, the present invention provides a series of coumarin derivatives containing thiazolidinedione structure, and determines their inhibitory activity against α-glucosidase. The results show that most of the synthesized derivatives exhibit significant inhibitory activity against α-glucosidase, especially the inhibition rates of compounds C15, C3 and C16 are 5.12±0.98μM, 6.58±0.75μM and 7.33±1.26μM, respectively, which are 34 to 128 times that of acarbose, among which C15 has the best inhibition rate, and its half inhibition concentration is shown in the figure below. Figure 1 The results showed that these compounds showed strong binding affinity when interacting with α-glucosidase. Therefore, compound C15 can be used as a lead compound for the development of α-glucosidase inhibitors with anti-diabetic activity.

[0158] The above is a detailed description of the embodiments of the present invention, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. A coumarin derivative containing a thiazolidinedione structure, characterized in that: The coumarin derivatives have the structural formula shown below: In the formula, R is selected from substituted or unsubstituted phenyl, and the substitution is optional.

2. The coumarin derivative containing a thiazolidinedione structure according to claim 1, characterized in that: The substitution is mono-substitution or poly-substitution; preferably, the substituent is selected from at least one of halogen, methyl, methoxy and hydroxyl.

3. 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:

4. A method for preparing a coumarin derivative containing a thiazolidinedione structure as claimed in any one of claims 1 to 3, the method comprising the following steps: Compound 2 and compound 5 containing an R group are used as raw materials to react to obtain the coumarin derivative containing a thiazolidinedione structure; Among them, the structures of compound 2 and compound 5 are as follows:

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

1.

6. The preparation method according to claim 4, characterized in that: The reaction temperature is 20-30° C.; preferably, the reaction time is 1-3 days; preferably, the reaction stirring speed is 400-600 rpm.

7. The preparation method according to claim 4, characterized in that: After the reaction is completed, a washing and filtering step is also included.

8. An α-glucosidase inhibitor, characterized in that The invention comprises the coumarin derivatives containing thiazolidinedione structure as claimed in any one of claims 1 to 3.

9. Use of the coumarin derivative containing a thiazolidinedione structure as claimed in any one of claims 1 to 3 in the preparation of a product for preventing and / or treating diabetes.

10. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the coumarin derivative containing a thiazolidinedione structure according to any one of claims 1 to 3 and / or a pharmaceutically acceptable salt thereof.

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