Hydrazinophenoxy acetic acid ethyl ester type alpha-glucosidase inhibitors, and preparation method and application thereof

Abstract

The application belongs to the field of biological medicine, and particularly relates to a hydrazine phenoxy ethyl acetate alpha-glucosidase inhibitor and a preparation method and application thereof. The inhibitor has the structure of formula (I). Compared with the prior art, the application further expands the inhibitor type with inhibitory effect on alpha-glucosidase activity, has excellent inhibitory activity, and has a simple preparation method, can be prepared on a large scale, and provides a new selection for different application requirements.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, and in particular relates to a hydrazine phenoxyethyl acetate α-glucosidase inhibitor, its preparation method and application. Background Technology

[0002] α-Glucosidase is an intestinal hydrolase that plays a key role in the degradation of carbohydrates into glucose and is one of the main targets for limiting blood sugar levels.

[0003] Currently, alpha-glucosidase inhibitors, such as acarbose, voglibose, and miglitol, are available clinically for the prevention and treatment of diabetes. Acarbose is currently the most widely used hypoglycemic drug for type 2 diabetes; however, long-term use of these drugs can cause significant side effects, including stomach pain, diarrhea, bloating, and allergic reactions. Furthermore, the raw materials for acarbose are extremely expensive, and the synthesis process is complex. Therefore, it is essential to find highly effective alpha-glucosidase inhibitors with fewer side effects for the treatment of type 2 diabetes.

[0004] Patent application CN202210670923.2 discloses an acridinone α-glucosidase inhibitor, which is extracted and prepared from the leaves of *Gnaphalium affine*. The inhibitor has two hydroxyl groups attached to its skeleton, and its structure is shown in Formula 1. The acridinone α-glucosidase inhibitor described in this invention exhibits good α-glucosidase inhibitory activity, IC50... 50 The value was 28.39 μM, significantly better than the positive control drug acarbose (IC50). 50 The value is 928.78 μM.

[0005]

[0006] Patent application CN202310472396.9 discloses a chalcone α-glucosidase inhibitor, its preparation method, and its application. The chalcone α-glucosidase inhibitor is prepared by introducing a phenyl group with hydroxyl and methoxy groups onto the chalcone skeleton, as shown in Formula 2. The optimal compound has an IC50 value of [missing information]. 50 The value was 239.7 ± 31.5 μM, which was superior to the positive control drug acarbose (IC50). 50 The value was 594.1 ± 3.2 μM.

[0007]

[0008] Patent application CN202010028778.9 discloses an α-glucosidase inhibitor and its application in hypoglycemic drugs. This invention discloses that dragon's blood stigmata extract A can inhibit α-glucosidase activity, and its inhibitory activity is far stronger than that of the clinical drug acarbose. The structural formula is shown in Formula 3 below. The IC50 of the compound... 50 The value is 16.03 ± 0.076 μM.

[0009]

[0010] As can be seen from the above, the existing technologies have a good inhibitory effect on α-glucosidase activity, but there is still room for further improvement. Moreover, the preparation process of the existing technologies is relatively complex, the separation method is complicated, and the yield is small. Therefore, it is necessary to propose different types of inhibitors that are easy to scale up and can meet different application needs. Summary of the Invention

[0011] The purpose of this invention is to solve at least one of the above-mentioned problems by providing a hydrazine phenoxyethyl α-glucosidase inhibitor, its preparation method, and its application. This invention further expands the types of inhibitors that inhibit α-glucosidase activity, exhibits excellent inhibitory activity, and has a simple and scalable preparation method, providing new options for different application needs.

[0012] The objective of this invention is achieved through the following technical solution:

[0013] The first aspect of this invention discloses a hydrazine phenoxyethyl acetate α-glucosidase inhibitor, wherein the inhibitor has the structure of formula (I):

[0014]

[0015] Preferably, the Ar group is selected from:

[0016]

[0017] Preferably, the inhibitor is:

[0018]

[0019] A second aspect of this invention discloses a method for preparing hydrazylphenoxyethyl acetate α-glucosidase inhibitors as described in any of the preceding embodiments, comprising the following steps:

[0020] S1: 1,5-Dichloro-2,4-dinitrobenzene is reacted with ethyl hydroxyacetate in a first organic solvent, and the first intermediate is obtained by separation;

[0021] S2: The first intermediate is reacted with hydrazine hydrate in a second organic solvent, and the second intermediate is obtained by separation;

[0022] S3: The second intermediate is reacted with the substituted acetophenone in a third organic solvent, filtered and dried to obtain the inhibitor.

[0023] Preferably, in step S1, the reaction uses triethylenediamine as a catalyst, K2CO3 as an acid-binding agent, and ethyl acetate as the first organic solvent; the reaction temperature is 50-80℃, preferably 75℃, and the reaction time is 14-48h, preferably 24h; the molar ratio of 1,5-dichloro-2,4-dinitrobenzene to ethyl hydroxyacetate is 90-120:90-120. More specifically, the ratio of the amount of 1,5-dichloro-2,4-dinitrobenzene, ethyl hydroxyacetate, triethylenediamine, anhydrous K2CO3, and the first organic solvent added is (90-120) mmol:(90-120) mmol:(1-4) mmol:(50-80) mmol:(190-220) mL, preferably 100 mmol:120 mmol:2 mmol:60 mmol:200 mL.

[0024] Preferably, in step S1, the separation is achieved by: filtration after the reaction is cooled (the filter cake can also be washed multiple times with small amounts of ethyl acetate and the filtrates combined to improve the yield), rotary evaporation of the filtrate under reduced pressure, ultrasonic dissolution in the first organic solvent, addition of petroleum ether to precipitate the first intermediate, filtration and drying to obtain the first intermediate.

[0025] Preferably, in step S2, the reaction uses ethyl acetate as the second organic solvent; the reaction temperature is 50-80℃, preferably 75℃, the reaction time is 1-4h, preferably 2h; and the molar ratio of the first intermediate to hydrazine hydrate is 20-30:21-31.5, preferably 20mmol:21mmol.

[0026] Preferably, in step S2, the separation is achieved by: filtration after the reaction is cooled (the filter cake can also be washed multiple times with small amounts of ethyl acetate and the filtrates combined to improve the yield), rotary evaporation of the filtrate under reduced pressure, and column chromatography separation to obtain the second intermediate.

[0027] Preferably, in step S3, the reaction uses hydrochloric acid as a catalyst and methanol as a third organic solvent; the reaction temperature is 50-65℃, preferably 65℃, and preferably heated in an oil bath; the reaction time is 1-4 hours, preferably 2 hours; the molar ratio of the second intermediate to the substituted acetophenone is 90-120:90-120. More specifically, the ratio of the second intermediate, the substituted acetophenone, and the hydrochloric acid added is (90-120) mmol:

[0028] (90-120)mmol:(5-15)mL, preferably 100mmol:100mmol:10mL.

[0029] The overall preparation process equation is shown below:

[0030]

[0031] In the equations, Equation I is the general structural formula of the inhibitor in this scheme, Equation II is the structural formula of the first intermediate, and Equation III is the structural formula of the second intermediate.

[0032] The third aspect of this invention discloses the use of a hydrazine phenoxyethyl α-glucosidase inhibitor as described in any of the preceding descriptions in the preparation of a drug that inhibits α-glucosidase activity.

[0033] This invention utilizes a receptor-based molecular docking virtual screening method to screen 400,000 compounds from the ZINC database, obtaining a compound that theoretically possesses α-glucosidase inhibitory activity. Subsequently, its structure was modified to design a more rational compound.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] This invention provides an α-glucosidase inhibitor with a novel skeletal structure, its preparation method, and its application. The synthesis method is simple, and the obtained inhibitor exhibits good α-glucosidase inhibitory activity and excellent α-glucosidase inhibitory effect. It can be applied to the preparation of drugs that inhibit α-glucosidase activity. Detailed Implementation

[0036] The present invention will now be described in detail with reference to specific embodiments, but these are by no means limitations on the present invention.

[0037] Unless otherwise specified, the reagents used in the following description may be commercially available products, and the methods used may be those known in the art.

[0038] A hydrazine phenoxyethyl acetate α-glucosidase inhibitor, said inhibitor having the structure shown in formula (I):

[0039]

[0040] Where Ar is selected from

[0041] Any one of them.

[0042] The above-mentioned method for preparing hydrazylphenoxyethyl acetate α-glucosidase inhibitors is illustrated by the following equation:

[0043]

[0044] The preparation method specifically includes the following steps:

[0045] (1) A reaction system was formed by 1,5-dichloro-2,4-dinitrobenzene and ethyl hydroxyacetate, and the intermediate of formula (II) was obtained after the reaction and treatment.

[0046] (2) Take the intermediate of formula (II) obtained in step (1) and dissolve it in an organic solvent to form a reaction system with hydrazine hydrate. After the reaction, the intermediate of formula (III) is obtained by treatment.

[0047] (3) Take the intermediate of formula (III) obtained in step (2) and the substituted acetophenone and dissolve them in an organic solvent to form a reaction system. After the reaction, the inhibitor shown in formula (I) is obtained by treatment.

[0048] In step (1), triethylenediamine is used as a catalyst, anhydrous K2CO3 is used as an acid-binding agent, and ethyl acetate is used as the organic solvent. The reaction system is placed at a temperature of 50-80℃, preferably 75℃, for a reaction time of 14-48h, preferably 24h. The post-treatment process is as follows: after the reaction is cooled to room temperature, the reaction system is filtered, the filter cake is washed with a small amount of ethyl acetate, the filtrates are combined, the filtrate is evaporated under reduced pressure to a solid state, a small amount of ethyl acetate is added, and the system is sonicated until it is homogeneous and free of large lumps. Add a large amount of petroleum ether to precipitate the product, filter and dry to obtain intermediate of formula (II). The ratio of the amount of 1,5-dichloro-2,4-dinitrobenzene, ethyl hydroxyacetate, triethylenediamine, anhydrous K2CO3 and organic solvent added is (90-120) mmol:(90-120) mmol:(1-4) mmol:(50-80) mmol:(190-220) mL, preferably 100 mmol:120 mmol:2 mmol:60 mmol:200 mL.

[0049] In step (2), the organic solvent is ethyl acetate, the reaction system is placed at a temperature of 50-80℃, preferably 75℃, and the reaction time is 1-4h, preferably 2h. The post-treatment process is as follows: after the material is completely converted, the temperature of the reaction system is reduced to room temperature for filtration. The filter cake is washed with a small amount of ethyl acetate, and the filtrate is evaporated under reduced pressure to obtain the crude product. The pure compound formula (III) intermediate is obtained by column chromatography. The additive ratio of formula (II) intermediate to hydrazine hydrate is (20-30) mmol:(21-31.5) mmol, preferably 20 mmol:21 mmol.

[0050] In step (3), hydrochloric acid is used as a catalyst, and anhydrous methanol is used as the organic solvent. The reaction system is placed in an oil bath and heated. The reaction temperature is 50-65℃, preferably 65℃, and the reaction time is 1-4h, preferably 2h. The post-treatment process is as follows: the reaction system is taken out and cooled. After the reaction solution is cooled to room temperature, it is filtered. The filter cake is washed with a small amount of anhydrous methanol and the filter cake is dried to obtain the inhibitor shown in formula (I). The ratio of the intermediate of formula (III), the substituted acetophenone and the hydrochloric acid is (90-120) mmol: (90-120) mmol: (5-15) mL, preferably 10 mmol: 10 mmol: 1 mL.

[0051] The prepared inhibitors were tested for their α-glucosidase inhibitory activity. The specific test method is as follows:

[0052] 1. Experimental instruments and materials

[0053] Multifunctional fluorescent microplate reader, SP-Max 3500FL, Shanghai Flash Spectrum Biotechnology Co., Ltd.;

[0054] Clean bench;

[0055] Bond A3 Pipette manual single-channel adjustable pipette, 0.5-10μL, 10-100μL, 100-1000μL, Titan Technology;

[0056] 96-well plate (white), sterilized, Corning;

[0057] α-Glucosidase was purchased from Beijing Innocare Technology Co., Ltd.; the fluorescent substrate p-nitrobenzene-α-D-glucopyranoside (PNPG) used in the enzyme inhibition experiment was purchased from Shanghai Myriel Biochemical Technology Co., Ltd.; disodium hydrogen phosphate dodecahydrate, disodium hydrogen phosphate dihydrate, and sodium carbonate were purchased from Titan Technology.

[0058] Positive control drug: acarbose, Shanghai Myrui Biochemical Technology Co., Ltd.

[0059] 2. Reagent preparation

[0060] a. 0.1 mol / L, pH 6.8 phosphate buffer (PBS): Mix 0.1 mM sodium dihydrogen phosphate solution with 0.1 mM disodium hydrogen phosphate solution and adjust the pH of the solution to 6.80.

[0061] b. Substrate solution: Accurately weigh 1.5 mg PNPG and dissolve it in 1 mL of PBS buffer to prepare a 5 mM PNPG solution. Prepare fresh before use.

[0062] c. Enzyme solution: Add 100U of lyophilized enzyme powder to PBS buffer to prepare a stock solution with a concentration of 10U / mL. Store at -20℃. Dilute to 0.4U / mL before use. Prepare fresh before use.

[0063] d. Termination solution: Weigh anhydrous sodium carbonate and dissolve it in distilled water to prepare a 0.2 mol / L sodium carbonate solution.

[0064] e. Positive control and sample solutions: Acarbose and ten target compounds were dissolved in DMSO to prepare an initial concentration of 3000 μmol / L. This was then serially diluted to eight concentration gradients: 2000 μmol / L, 1000 μmol / L, 500 μmol / L, 250 μmol / L, 125 μmol / L, 62.5 μmol / L, 31.25 μmol / L, and 15.625 μmol / L. Three sets of each concentration gradient were prepared sequentially.

[0065] 3. Experimental Methods

[0066] This experiment was conducted in a 96-well plate with a total reaction volume of 200 μL. 100 μL of PBS buffer was added to each well, followed by 20 μL of different concentrations of the α-glucosidase inhibitor sample or positive control sample solution. Then, 20 μL of α-glucosidase solution was added, and the plate was incubated at 37°C for 30 min. Next, 20 μL of PNPG solution was added, and incubation continued for 15 min. Finally, 40 μL of sodium carbonate solution was added to each well to terminate the reaction. The absorbance of each well was measured at 405 nm. The experiment was performed in triplicate.

[0067] The experiment consisted of four groups:

[0068] Sample group A (enzyme solution + substrate solution + buffer solution + test sample solution / acarbose solution + stop solution);

[0069] Sample background group A0 (sample solution + substrate solution + buffer solution + stop solution);

[0070] Negative control group B (enzyme solution + substrate solution + buffer + DMSO + stop solution);

[0071] Negative background control group B0 (substrate solution + buffer solution + DMSO + stop solution);

[0072] The amount of reagents added to each group is shown in Table 1.

[0073] Table 1: Amount of reagents added to the four experimental groups

[0074]

[0075] After measuring the absorbance, the corresponding inhibition rate was calculated using a formula. The data was then processed using GraphPad Prism software to fit a curve and the corresponding IC. 50 Value. The formula for the inhibition rate of the sample against α-glucosidase is as follows:

[0076]

[0077] A represents the absorbance of the substrate and α-glucosidase after incubation at 37°C for 30 min in the presence of the sample solvent.

[0078] A0 represents the background absorbance of the system after 30 minutes of reaction in the presence of the sample and solvent without the addition of α-glucosidase.

[0079] B represents the absorbance of the substrate and α-glucosidase after 30 minutes of incubation.

[0080] B0 represents the absorbance after incubation for 30 minutes with only the substrate and solvent system added.

[0081] Example 1

[0082] (Z)-2-(2,4-dinitro-5-(2-(1-(2,4,6-trimethoxyphenyl)ethylene)hydrazine)phenoxy)ethyl acetate, the structural formula of which is shown below:

[0083]

[0084] The specific synthesis steps are as follows:

[0085] Accurately weigh 23.70 g (100 mmol) of 1,5-dichloro-2,4-dinitrobenzene, 12.49 g (120 mmol) of ethyl hydroxyacetate, 0.2242 g (2 mmol) of triethylenediamine and 8.29 g (60 mmol) of anhydrous K2CO3, and add them sequentially to a round-bottom flask. Pour in 200 mL of ethyl acetate and place in an oil bath at 75 °C. Stir and react for 24 hours. After the reaction is complete, cool the reaction system to room temperature and filter the system. Wash the filter cake with a small amount of ethyl acetate, combine the filtrates, and evaporate the filtrate under reduced pressure to obtain a solid. Add a small amount of ethyl acetate and sonicate until the system is homogeneous and free of large lumps. Then add a large amount of petroleum ether to precipitate the product. Filter and dry to obtain intermediate (II).

[0086] Accurately weigh 6.09 g (20 mmol) of intermediate (II) and 1.05 g (21 mmol) of hydrazine hydrate into a 50 mL round-bottom flask. Add 20 mL of ethyl acetate and heat to dissolve. Then, use a 5 mL disposable syringe to add 1.05 g (21 mmol) of hydrazine hydrate into the system and react at 75 °C for 2 hours. After the reaction is complete, cool the reaction system to room temperature and filter. Wash the filter cake with a small amount of ethyl acetate. Distill the filtrate under reduced pressure to obtain the crude product, which is then separated by column chromatography to obtain the pure intermediate (III).

[0087] Accurately weigh 3 g (10 mmol) of intermediate (III) and 2.1 g (10 mmol) of 2,4,6-trimethoxyacetophenone into a 50 mL round-bottom flask, add 10 mL of anhydrous methanol, and then add 1 mL of hydrochloric acid dropwise to the system using a 2 mL disposable syringe. React at 65 °C for 2 hours. After the reaction is complete, remove the reaction system and cool it. After the reaction solution cools to room temperature, filter it. Wash the filter cake with a small amount of anhydrous methanol and dry the filter cake to obtain the inhibitor shown in formula (I).

[0088] Experimental results

[0089] (Z)-2-(2,4-dinitro-5-(2-(1-(2,4,6-trimethoxyphenyl)ethylene)hydrazyl)phenoxy)ethyl acetate, pale yellow powder / solid, 95% yield, IC50 50 The value was 11.62±2.6 μM, and the IC50 of the positive control drug was... 50 The value is 11.14 ± 1.8 μM.

[0090] 1 H NMR(401MHz,DMSO-d6)δ10.97(s,1H),8.85(s,1H),7.07(s,1H),6.31(s,2H),5.06(s,2 H), 4.11 (q, J = 7.1Hz, 2H), 3.83 (s, 3H), 3.76 (s, 6H), 2.22 (s, 3H), 1.08 (t, J = 7.1Hz, 3H). 13 C NMR(101MHz,DMSO-d6)δ161.98,158.84,156.87,145.35,130.09,127.95,124.38,110.5 8,92.46,90.87,66.56,61.59,58.57,57.13,56.65,55.70,55.22,19.51,13.61,12.33.

[0091] Example 2

[0092] (Z)-2-(2,4-dinitro-5-(2-(1-(4-chlorophenyl)ethylene)hydrazyl)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared in a manner similar to Example 1.

[0093]

[0094] Orange powdery solid, 90% yield, IC 50 The value was 27.04 ± 3.9 μM.

[0095] 1 H NMR (401MHz, DMSO-d6) δ11.00(s,1H),8.82(s,1H),7.86(d,J=8.3Hz,2H),7.29(s,1H),6.98(d,J=8.4Hz,2H),5.2 1(s,2H),4.22(q,J=7.0Hz,2H),4.09(q,J=7.0Hz,2H),2.38(s,3H),1.36(t,J=6.9Hz,3H),1.23(p,J=6.0Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.83,156.94,153.20,145.22,130.01,129.64,129.32,127.75, 126.23,124.35,115.61,113.98,97.45,68.21,66.69,15.65,14.44,14.32,13.87,13.07.

[0096] Example 3

[0097] (Z)-2-(2,4-dinitro-5-(2-(1-(4-methoxy-3-nitrophenyl)ethylene)hydrazyl)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared in a manner similar to Example 1.

[0098]

[0099] Yellow powdery solid, yield 88%, IC 50 The value was 34.41 ± 7.4 μM.

[0100] 1H NMR (401MHz, DMSO-d6) δ11.03(s,1H),8.82(s,1H),8.34(d,J=2.3Hz,1H),8.20(dd,J=8.9,2.4Hz,1H),7.43(d, J=9.0Hz,1H),7.34(s,1H),5.24(s,2H),4.20(q,J=7.1Hz,2H),4.00(s,3H),2.43(s,3H),1.18(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.84,156.96,153.23,151.18,145.03,140.00,133.15,130.44,129.91,127.86,1 26.10,124.64,115.60,99.49,97.76,66.75,63.22,61.75,58.23,56.79,55.35,15.09,14.35,13.79,13.10.

[0101] Example 4

[0102] (Z)-2-(2,4-dinitro-5-(2-(1-(4-chlorophenyl)ethylene)hydrazyl)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared in a manner similar to Example 1.

[0103]

[0104] Orange powdery solid, 94% yield, IC 50 The value is 56.08±2.4μM.

[0105] 1 H NMR(401MHz, DMSO-d6)δ11.01(s,1H),8.81(d,J=1.4Hz,1H),7.99–7.89(m,2H),7.54–7.47(m, 2H),7.31(s,1H),5.22(s,2H),4.21(q,J=7.1Hz,2H),2.40(s,3H),1.20(td,J=7.1,1.3Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.81,156.92,145.10,136.31,135.26,130.42,129.92,129.51,129.45, 128.55,128.16,127.89,126.08,124.63,97.74,66.76,61.77,60.30,15.15,14.40,13.85,13.15.

[0106] Example 5

[0107] (Z)-2-(2,4-dinitro-5-(2-(1-(4-aminosulfonylphenyl)ethylene)hydrazyl)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared in a manner similar to Example 1.

[0108]

[0109] Yellow powdery solid, yield 93%, IC 50 The value was 78.83±23.8μM.

[0110] 1 H NMR (401MHz, DMSO-d6) δ11.08(d,J=2.5Hz,1H),8.85(d,J=2.7Hz,1H),8.11(dd,J=8.4,2.7Hz,2H),7.91(dd,J=8.7,2.7Hz,2H),7.51(d,J =2.6Hz,2H),7.39(d,J=2.7Hz,1H),5.27(d,J=2.6Hz,2H),4.22(td,J=7.2,2.7Hz,2H),2.48(d,J=2.7Hz,3H),1.21(td,J=7.1,2.7Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.89,156.96,151.83,145.32,145.16,140.66,130.64,128.26, 127.85,127.16,126.71,125.50,124.90,99.74,68.35,66.83,15.80,15.21,14.57,13.91.

[0111] Example 6

[0112] (Z)-2-(2,4-dinitro-5-(2-(1-(4-cyanophenyl)ethylene)hydrazyl)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared in a manner similar to Example 1.

[0113]

[0114] Yellow powdery solid, yield 93%, IC 50 The value was 83.31 ± 16.3 μM.

[0115] 1H NMR (401MHz, DMSO-d6) δ11.07(s,1H),8.83(s,1H),8.09(d,J=8.2Hz,2H),7.93(d,J=8.2Hz ,2H),7.35(s,1H),5.26(s,2H),4.22(q,J=7.1Hz,2H),2.44(s,3H),1.19(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.83,156.92,151.34,144.98,141.77,133.78,132.12,130.79,128.47, 127.76,126.01,124.96,119.10,112.44,99.79,68.33,66.82,61.76,15.17,14.38,13.89,13.12.

[0116] Example 7

[0117] (Z)-2-(2,4-dinitro-5-(2-(1-(p-tolyl)ethylene)hydrazine)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared using a method similar to Example 1.

[0118]

[0119] Orange powdery solid, yield 91%, IC 50 The value was 93.83±14.7μM.

[0120] 1 H NMR (401MHz, DMSO-d6) δ10.99(s,1H),8.81(s,1H),7.80(d,J=8.0Hz,2H),7.32–7.24(m ,3H),5.21(s,2H),4.23(q,J=7.1Hz,2H),2.37(d,J=6.4Hz,6H),1.23(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.81,156.92,153.25,145.23,140.36,134.67,130.15,128.81,127.66 ,127.61,126.16,124.44,97.55,68.21,66.69,61.81,21.99,20.75,15.16,14.35,13.87,13.09.

[0121] Example 8

[0122] (Z)-2-(5-(2-(1-(2-hydroxyphenyl)ethylene)hydrazyl)-2,4-dinitrophenoxy)ethyl acetate, with the following structural formula, was prepared using a method similar to Example 1.

[0123]

[0124] Orange powdery solid, yield 89%, IC 50 The value is 119.8 ± 22.9 μM.

[0125] 1 H NMR (401MHz, DMSO-d6) δ11.06(s,1H),10.75(s,1H),8.86(s,1H),7.59–7.53(m,1H),7.33(t,J=7.5Hz,1H),7.06(s,1H ), 6.94(dd,J=17.0,8.1Hz,2H),5.18(s,2H),4.22(q,J=7.1Hz,2H),2.52(dd,J=3.7,1.9Hz,2H),1.20(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.61,157.79,157.22,144.85,132.69,131.04,130.26,127.95, 124.60,123.47,118.89,118.81,116.33,68.05,66.52,61.79,18.01,16.77,15.04,13.76.

[0126] Example 9

[0127] (Z)-2-(5-(2-(1-(2,5-dihydroxyphenyl)ethylene)hydrazine)-2,4-dinitrophenoxy)ethyl acetate, with the following structural formula, was prepared using a method similar to Example 1.

[0128]

[0129] Orange powdery solid, yield 91%, IC 50 The value is 124.3±36.8μM.

[0130] 1H NMR (401MHz, DMSO-d6) δ11.05(s,1H),10.11(s,1H),9.00(s,1H),8.87(s,1H),7.02(s,1H),6.92(t,J=1 .7Hz,1H),6.77(d,J=1.7Hz,2H),5.16(s,2H),4.23(q,J=7.1Hz,2H),2.45(s,3H),1.21(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.56,157.88,157.02,150.22,149.89,144.79,130.25,127.98,126.25,124.62,123 .29,119.78,118.57,118.18,116.96,116.02,114.44,98.89,97.22,68.04,66.51,61.84,15.40,15.00,13.72.

[0131] Example 10

[0132] (Z)-2-(2,4-dinitro-5-(2-(1-(4-nitrophenyl)ethylene)hydrazyl)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared in a manner similar to Example 1.

[0133]

[0134] Yellow powdery solid, 94% yield, IC 50 The value is 155.5 ± 42.9 μM.

[0135] 1 H NMR (401MHz, DMSO-d6) δ11.11(s,1H),8.85(s,1H),8.29(d,J=8.7Hz,2H),8.19(d,J=8.6Hz,2 H),7.40(s,1H),5.28(s,2H),4.22(q,J=7.1Hz,2H),2.53(s,2H),1.21(q,J=8.7,7.1Hz,3H). 13 C NMR (101MHz, DMSO-d6) δ167.86,156.93,144.98,130.92,128.86,127.77,125.11,124.97,123.24,99.93,14.55.

[0136] Example 11

[0137] (Z)-2-(5-(2-(1-(3-hydroxyphenyl)ethylene)hydrazyl)-2,4-dinitrophenoxy)ethyl acetate, with the following structural formula, was prepared using a method similar to Example 1.

[0138]

[0139] Orange powdery solid, yield 93%, IC 50 The value is 195.6 ± 58.4 μM.

[0140] 1 H NMR (401MHz, DMSO-d6) δ11.01(s,1H),9.65(s,1H),8.83(s,1H),7.34(d,J=7.9Hz,1H),7.31–7.22(m,3 H), 6.89 (dd, J = 8.1, 2.2Hz, 1H), 5.20 (s, 2H), 4.23 (q, J = 7.1Hz, 2H), 2.38 (s, 3H), 1.22 (t, J = 7.1Hz, 3H). 13 C NMR(101MHz,DMSO-d6)δ167.74,158.03,156.92,153.42,145.25,138.91,130.78,130.23,129.12,127.92,126.18,124 .55,124.49,118.56,118.18,116.90,116.60,113.01,97.57,68.17,66.65,63.31,60.33,15.09,14.66,13.80,13.40.

[0141] Example 12

[0142] (Z)-2-(5-(2-(1-(3,4-difluorophenyl)ethylene)hydrazyl)-2,4-dinitrophenoxy)ethyl acetate, with the following structural formula, was prepared using a method similar to Example 1.

[0143]

[0144] Pale yellow powdery solid, yield 93%, IC 50 The value was 201.4 ± 57 μM.

[0145] 1H NMR (401MHz, DMSO-d6) δ11.01(s,1H),8.83(s,1H),8.04–7.95(m,1H),7.76(s,1H),7.54(q,J=9 .0Hz,1H),7.35(s,1H),5.28(s,2H),4.22(q,J=7.1Hz,2H),2.42(s,3H),1.20(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.89,156.89,149.71,145.12,145.05,135.17,130.52,127.79,126 .05,125.04,124.72,118.86,99.62,97.91,68.30,66.78,61.72,15.05,14.41,13.77,13.15.

[0146] Example 13

[0147] (Z)-2-(2,4-dinitro-5-(2-(1-(o-tolyl)ethylene)hydrazine)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared in a manner similar to Example 1.

[0148]

[0149] Orange-yellow powdery solid, yield 92%, IC 50 The value is 215.5 ± 42.5 μM.

[0150] 1 H NMR(401MHz, DMSO-d6)δ11.00(s,1H),8.86(s,1H),7.44(d,J=7.3Hz,1H),7.39–7.26(m,3H), 7.17(s,1H),5.14(s,2H),4.14(q,J=7.1Hz,2H),2.45(d,J=2.1Hz,6H),1.12(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.63,156.93,156.13,145.38,138.75,135.93,130.72,130.31,130.27,130.16,129.70,128.48,1 27.94,125.57,124.48,124.43,97.46,68.01,67.85,66.46,61.71,60.24,21.98,20.73,18.41,17.15,14.98,13.70,12.42.

[0151] Example 14

[0152] (Z)-2-(5-(2-(1-(4-aminophenyl)ethylene)hydrazyl)-2,4-dinitrophenoxy)ethyl acetate, with the following structural formula, was prepared using a method similar to Example 1.

[0153]

[0154] Dark purple powdery solid, yield 93%, IC 50 The value is 225.2 ± 50.5 μM.

[0155] 1 H NMR (401MHz, DMSO-d6) δ11.01(s,1H),8.82(s,1H),7.65(d,J=8.5Hz,2H),7.26(s,1H),6.62(d,J =8.5Hz,2H),5.69(s,2H),5.20(s,2H),4.23(q,J=7.1Hz,2H),2.32(s,3H),1.25(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.88,156.91,154.11,151.60,151.52,145.07,129.62,129.19,129.12,1 27.70,126.40,124.19,124.06,114.52,114.47,112.98,97.19,68.15,66.64,61.79,15.23,14.00.

[0156] Example 15

[0157] (Z)-2-(5-(2-(1-(4-iodophenyl)ethylene)hydrazyl)-2,4-dinitrophenoxy)ethyl acetate, with the following structural formula, was prepared in a manner similar to Example 1.

[0158]

[0159] Orange powdery solid, 90% yield, IC 50 The value is 351.4 ± 75.4 μM.

[0160] 1H NMR (401MHz, DMSO-d6) δ11.03(s,1H),8.84(s,1H),7.83(d,J=8.4Hz,2H),7.71(d,J=8.4Hz ,2H),7.32(s,1H),5.24(s,2H),4.21(q,J=7.1Hz,2H),2.41(s,3H),1.20(t,J=7.1Hz,3H). 13 C NMR (101MHz, DMSO-d6) δ167.85,145.17,138.70,137.06,130.46,129.66,128.10,127.87,124.70,97.82,97.47,66.78,14.35,13.09.

[0161] Example 16

[0162] (Z)-2-(2,4-dinitro-5-(2-(1-(tert-butyl)ethylene)hydrazine)phenoxy)ethyl acetate, the structural formula of which is shown below, was prepared in a manner similar to Example 1.

[0163]

[0164] Orange-yellow powdery solid, yield 90%, IC 50 The value is 514.1 ± 86.8 μM.

[0165] 1 H NMR (401MHz, DMSO-d6) δ11.01(s,1H),8.81(s,1H),7.86–7.79(m,2H),7.47(d,J=8.3Hz,2H),7. 31(s,1H),5.20(s,2H),4.22(q,J=7.1Hz,2H),2.39(s,3H),1.33(s,9H),1.22(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ167.79,156.91,153.26,145.26,134.77,130.15,127.57,127.51,126.62,126.56,126.15,1 26.00,125.94,124.94,124.45,97.59,68.19,66.67,61.77,34.98,32.00,30.80,29.61,15.10,14.42,13.81,13.16.

[0166] Example 17

[0167] (Z)-2-(5-(2-(1-(4-bromophenyl)ethylene)hydrazyl)-2,4-dinitrophenoxy)ethyl acetate, with the following structural formula, was prepared using a method similar to Example 1.

[0168]

[0169] Orange powdery solid, 94% yield, IC 50 The value is 596.1±91μM.

[0170] 1 H NMR (401MHz, DMSO-d6) δ11.04(s,1H),8.85(s,1H),7.88(d,J=8.3Hz,2H),7.67(d,J=8.3Hz,2H), 7.34(s,1H),5.25(s,2H),4.22(q,J=7.1Hz,2H),2.52(s,1H),2.43(s,3H),1.20(t,J=7.1Hz,3H). 13 C NMR (101MHz, DMSO-d6) δ167.85,156.93,145.16,132.75,131.19,130.48,129.78,128.17,127.87,124.73,99.57,97.84,61.75,13.19.

[0171] Example 18

[0172] (Z)-2-(5-(2-(1-(2-hydroxy-4-methoxyphenyl)ethylene)hydrazyl)-2,4-dinitrophenoxy)ethyl acetate, with the following structural formula, was prepared in a manner similar to Example 1.

[0173]

[0174] Red powdery solid, 90% yield, IC 50 The value is 600.5 ± 75.6 μM.

[0175] 1 H NMR (401MHz, DMSO-d6) δ11.40(s,1H),11.05(s,1H),8.86(s,1H),7.58(d,J=8.8Hz,1H),6.91(s,1H),6 .57–6.47(m,2H),5.18(s,2H),4.25(q,J=7.1Hz,2H),3.79(s,3H),2.48(s,3H),1.24(t,J=7.1Hz,3H). 13C NMR(101MHz,DMSO-d6)δ167.57,159.61,159.21,157.06,144.55,130.10,128.05,126.32,124.52,114 .95,107.34,105.74,102.75,101.18,96.95,67.99,66.47,56.53,55.09,16.09,15.06,14.83,13.76.

[0176] In summary, α-glucosidase assays were performed on 18 compounds, with acarbose as a positive control. The IC50 of acarbose was... 50 The value is 11.14 ± 1.8 μM.

[0177] The IC50 of the following five compounds synthesized in this invention 50 The values ​​are all close to 11.14 ± 1.8 μM:

[0178]

[0179] Among them, the compound with the best inhibitory effect Its IC 50 The value was 11.62±2.6 μM, indicating excellent α-glucosidase inhibitory activity.

[0180] In summary, this method proposes a novel α-glucosidase inhibitor with a simple preparation method, readily available raw materials, and low production cost. Furthermore, its inhibitory effect on α-glucosidase activity is comparable to that of acarbose, demonstrating broad application prospects.

[0181] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. A hydrazine phenoxyethyl acetate α-glucosidase inhibitor, characterized in that, The inhibitor mentioned is: , , , , , or .

2. A method for preparing the hydrazylphenoxyethyl α-glucosidase inhibitor as described in claim 1, characterized in that, Includes the following steps: S1: 1,5-Dichloro-2,4-dinitrobenzene is reacted with ethyl hydroxyacetate in a first organic solvent, and the first intermediate is obtained by separation; S2: The first intermediate is reacted with hydrazine hydrate in a second organic solvent, and the second intermediate is obtained by separation; S3: The second intermediate is reacted with the substituted acetophenone in a third organic solvent, filtered and dried to obtain the inhibitor.

3. The method for preparing a hydrazine phenoxyethyl acetate α-glucosidase inhibitor according to claim 2, characterized in that, In step S1, the reaction uses triethylenediamine as a catalyst, K2CO3 as an acid-binding agent, and ethyl acetate as the first organic solvent; the reaction temperature is 50-80℃, and the reaction time is 14-48h; the molar ratio of 1,5-dichloro-2,4-dinitrobenzene to ethyl hydroxyacetate is 90-120:90-120.

4. The method for preparing a hydrazine phenoxyethyl acetate α-glucosidase inhibitor according to claim 2, characterized in that, In step S1, the separation is achieved by: filtering after the reaction is cooled, taking the filtrate and rotary evaporating under reduced pressure, dissolving it in the first organic solvent by sonication, adding petroleum ether to precipitate the first intermediate, filtering and drying to obtain the first intermediate.

5. The method for preparing a hydrazine phenoxyethyl acetate α-glucosidase inhibitor according to claim 2, characterized in that, In step S2, the reaction uses ethyl acetate as the second organic solvent; the reaction temperature is 50-80℃, the reaction time is 1-4h; and the molar ratio of the first intermediate to hydrazine hydrate is 20-30:21-31.

5.

6. The method for preparing a hydrazine phenoxyethyl acetate α-glucosidase inhibitor according to claim 2, characterized in that, In step S2, the separation is achieved by: filtering after the reaction is cooled, taking the filtrate and evaporating under reduced pressure, and then separating by column chromatography to obtain the second intermediate.

7. The method for preparing a hydrazine phenoxyethyl acetate α-glucosidase inhibitor according to claim 2, characterized in that, In step S3, the reaction uses hydrochloric acid as a catalyst and methanol as the third organic solvent; the reaction temperature is 50-65℃ and the reaction time is 1-4h; the molar ratio of the second intermediate to the substituted acetophenone is 90-120:90-120.

8. The use of the hydrazine phenoxyethyl α-glucosidase inhibitor as described in claim 1 in the preparation of a medicament for inhibiting α-glucosidase activity.

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