1, 2, 4-triazole compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicine synthesis, and particularly relates to a 1, 2, 4-triazole compound as well as a preparation method and application thereof. The 1, 2, 4-triazole compound provided by the invention has a structure as shown in a formula I: # imgabs0 # test results show that the 1, 2, 4-triazole compound provided by the invention not only has a better selective inhibition effect on HDAC6 protein, but also can inhibit proliferation of gastric adenocarcinoma cells, has a protection effect on myocardial cells, and can be used for preparing a medicine for treating gastric adenocarcinoma cells. The compound can be developed and applied as an HDAC6 inhibitor, an anti-gastric cancer drug or a myocardial protection drug. Therefore, the skeleton type of the compound in an HDAC6 inhibitor is enriched, and meanwhile, a new direction and data support are provided for developing an anti-tumor drug which is low in myocardial injury risk and has HDAC6 selectivity.

Description

Technical Field

[0001] The present invention belongs to the technical field of drug synthesis, and in particular relates to a 1,2,4-triazole compound and a preparation method and application thereof. Background Art

[0002] Cancer remains a leading cause of death in my country. While improvements in diagnosis and treatment have led to improved cancer survival rates, the risk of adverse reactions has also increased. Currently, cancer treatment primarily involves surgery, radiotherapy, and chemotherapy. While these approaches can achieve certain therapeutic effects, they can also lead to cardiotoxicity, including heart failure, left ventricular dysfunction, coronary artery disease, and myocarditis. Because both traditional and novel cancer therapies can have cardiovascular toxicity, minimizing these side effects is becoming increasingly important as long-term cancer survival rates improve.

[0003] Molecular targeted therapy refers to the precise attack on cancer cells by specifically blocking key molecules related to tumor growth and metastasis (such as tyrosine kinase). Compared with traditional therapies, molecular targeted therapy can focus on tumor-specific abnormal molecules and cause less damage to normal cells. Therefore, the rational use of molecular targeted therapy can reduce the toxic side effects of tumor treatment and improve the efficacy. Histone deacetylase 6 (HDAC6) is a member of the class IIb histone deacetylase family, which is different from other Zn 2+ Dependent HDACs have obvious uniqueness. HDAC6 is located in the cytoplasm and contains two catalytic domains (CD1 and CD2) and a ubiquitin-binding zinc finger domain in its structure. Due to the unique structure, substrate and biological function of HDAC6, targeted inhibition of its deacetylase activity has become an important strategy for innovative drug development, including cancer, neurodegenerative diseases, cardiovascular diseases and immunological diseases. At present, researchers have developed a variety of HDAC6 selective inhibitors with different surface recognition regions (Cap), linkers and zinc ion binding groups (ZBG) through more than 20 years of comprehensive efforts. However, due to the limited clinical efficacy, acquired drug resistance and the inability to inhibit the non-enzymatic functions of HDAC6, traditional HDAC6 inhibitors are hindered, which limits their further development and application in clinical practice.

[0004] Therefore, exploring and developing a new generation of small molecule compounds with low cytotoxicity, high HDAC6 inhibitory activity and subtype selectivity is of great significance for the treatment of cancer and other related diseases. Summary of the Invention

[0005] To address the above issues, the present invention aims to provide a 1,2,4-triazole compound, its preparation method, and its application. The 1,2,4-triazole compound provided by the present invention not only has excellent selective inhibition of HDAC6, but also has good anti-proliferative ability against gastric adenocarcinoma cells and a certain protective effect on cardiomyocytes.

[0006] To achieve the above objectives, the first aspect of the present invention provides a 1,2,4-triazole compound having a structure shown in Formula I:

[0007]

[0008] In formula I, R1 is H, -Ph-4-CH3, -Ph-3-CH3, -Ph-2-CH3, -Ph-4-C2H5, -Ph-4-C3H7, -Ph-4-OCH3, -Ph-3,4,5-OCH3, -Ph-4-Cl, -Ph-3-Cl, -Ph-4-F, -Ph-4-Br, -Ph-4-NO2, One of the following;

[0009] R2 is one of H, 4-CH3, 3-CH3, 2-CH3, 4-C3H7, 4-C4H9, 4-OCH3, 3,4-CH3, 2,4-CH3, 4-F, 3-F, 3-F, 3-Cl, 4-Cl, and 4-Br;

[0010] R3 is One of OH.

[0011] Preferably, the 1,2,4-triazole compound is selected from the following compounds, which are sequentially denoted as compounds Y-1 to Y-47:

[0012]

[0013]

[0014]

[0015] The second aspect of the present invention provides a method for preparing the above-mentioned 1,2,4-triazole compounds, using the following technical route:

[0016]

[0017] When R3 is OH, the preparation method of the 1,2,4-triazole compound comprises the following steps:

[0018] Raw materials A and B are dissolved in solvent a, and heated under reflux to react to obtain intermediate compound C; intermediate compound C is dissolved in solvent b, and then reacted with raw material D, tert-butyl hydroperoxide, I2, and an alkaline substance, followed by column chromatography to obtain intermediate compound E; intermediate compound E is dissolved in solvent c, and then reacted with an alkaline substance to obtain intermediate compound F; intermediate compound F is dissolved in solvent d, and then reacted with raw material G and a condensation reagent to obtain intermediate compound H; intermediate compound H is dissolved in solvent e, and then reacted with an alkaline substance to obtain compound I, which is the 1,2,4-triazole compound;

[0019] When R3 is When the 1,2,4-triazole compound is prepared, the method comprises the following steps:

[0020] Raw materials A and B are dissolved in solvent a and subjected to heating reflux reaction to obtain intermediate compound C; intermediate compound C is dissolved in solvent b, and then reacted with raw material D, tert-butyl hydroperoxide, I2, and an alkaline substance, followed by column chromatography to obtain intermediate compound E; intermediate compound E is dissolved in solvent c, and then reacted with an alkaline substance to obtain intermediate compound F; intermediate compound F is dissolved in solvent d, and then reacted with raw material G and a condensation reagent to obtain intermediate compound H; intermediate compound H is dissolved in solvent e, and then reacted with an alkaline substance to obtain compound I; compound I is added to solvent f, and then reacted with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine and a condensation reagent, followed by column chromatography to obtain intermediate compound J; intermediate compound J is dissolved in solvent g, and then reacted with an acidic substance, and then filtered to obtain compound K, which is the 1,2,4-triazole compound;

[0021] When R3 is When the 1,2,4-triazole compound is prepared, the method comprises the following steps:

[0022] The raw materials A and B are dissolved in solvent a and heated under reflux to react to obtain an intermediate compound C; the intermediate compound C is dissolved in solvent b, and then reacted with raw material D, tert-butyl hydroperoxide, I2, and an alkaline substance, followed by column chromatography to obtain an intermediate compound E; the intermediate compound E is dissolved in solvent c, and then reacted with an alkaline substance to obtain an intermediate compound F; the intermediate compound F is dissolved in solvent d, and then reacted with raw material G and a condensation reagent to obtain an intermediate compound H; the intermediate compound H is dissolved in solvent e, and then reacted with an alkaline substance to obtain compound I; compound I is dissolved in solvent h, and then reacted with Boc-hydrazine and a condensation agent to obtain an intermediate compound L; the intermediate compound L is dissolved in solvent i, and then reacted with an acidic substance, and then filtered to obtain compound M, which is the 1,2,4-triazole compound.

[0023] Preferably, the alkaline substance is triethylamine or lithium hydroxide.

[0024] Preferably, the condensation reagent is one or more of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate, 1-methylimidazole, N,N-diisopropylethylamine, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate.

[0025] Preferably, the acidic substance is trifluoroacetic acid or hydrochloric acid.

[0026] Preferably, the solvent is one or more of ethanol, acetonitrile, methanol, dichloromethane and water.

[0027] The third aspect of the present invention provides the use of the above-mentioned 1,2,4-triazole compounds, which is the use of the 1,2,4-triazole compounds in the preparation of HDAC6 inhibitors and / or anti-gastric cancer drugs and / or myocardial protective drugs.

[0028] Preferably, the anti-gastric cancer drug is a drug that inhibits the proliferation activity of gastric cancer cells; and the myocardial protective drug is a drug that enhances the activity of myocardial cells.

[0029] Preferably, the gastric cancer cells are AGS cells; and the cardiomyocytes are H9c2 cells.

[0030] The technical solution of the present invention has the following advantages and beneficial effects:

[0031] The 1,2,4-triazole compounds provided by the present invention have novel structures. Experimental results show that the 1,2,4-triazole compounds of the present invention not only have a relatively good selective inhibitory effect on HDAC6 protein, but also can inhibit the proliferation of gastric adenocarcinoma cells, while having a protective effect on myocardial cells, and can be developed and applied as HDAC6 inhibitors, anti-gastric cancer drugs or myocardial protective drugs. The preparation method of the above-mentioned compounds provided by the present invention has mild synthesis conditions and is easy to implement, and the compounds obtained by the method have the characteristics of high yield and high purity. Therefore, the present invention has good application prospects in the preparation of inhibitors based on the HDAC6 target and the development of anti-tumor drugs with low myocardial damage risk and selectivity. At the same time, it is of great significance to the development of drugs based on the HDAC6 target and the treatment of related diseases. DETAILED DESCRIPTION

[0032] The technical solution of the present invention is described in detail below in conjunction with specific implementation methods.

[0033] Example 1

[0034] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-1, having the structural formula: The preparation route and preparation process of the compound are as follows:

[0035]

[0036] (1) Benzaldehyde (1.2 mmol) and phenylhydrazine (1.0 mmol) were placed in a 250 mL round-bottom flask, and 100 mL of 20% ethanol aqueous solution (ethanol: water = 2:8, v / v) was added to dissolve the mixture. The mixture was reacted in an oil bath at 80°C for 6-8 h and monitored by TLC (petroleum ether: ethyl acetate = 20:1). After the reaction was completed, the mixture was directly filtered while hot. The filter cake was rinsed with ethyl acetate in small amounts several times and then dried to obtain intermediate a.

[0037] (2) After dissolving intermediate a (1 mmol) in 100 mL of acetonitrile, glycine methyl ester hydrochloride (1.5 mmol), tert-butyl hydroperoxide (TBHP, 1.5 mmol), I2 (1.4 mmol), and triethylamine (0.2 mmol) were added and reacted in an oil bath at 90°C for 3 h. The mixture was monitored by TLC (petroleum ether: ethyl acetate = 10:1), extracted three times with ethyl acetate and water, and the upper organic phases were combined and washed with saturated brine. The organic phase was removed by rotary evaporation and subjected to column chromatography (eluents: ethyl acetate and petroleum ether) to obtain intermediate b.

[0038] (3) Intermediate b (1 mmol) was dissolved in 25 mL of methanol, and lithium hydroxide (10 mmol) was added and hydrolyzed at 70°C for 2 h. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 3:1). After the reaction, the reaction solution was dried to obtain intermediate c.

[0039] (4) The intermediate c (1 mmol) obtained in the previous step was dissolved in 25 mL of acetonitrile with methyl para-aminobenzoate (1.2 mmol) and 1-methylimidazole (2 mmol). After stirring for 5 min, N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol) was added and reacted at room temperature for 3 h. After monitoring by TLC (petroleum ether:ethyl acetate = 3:1), the intermediate d was obtained by filtration.

[0040] (5) The dried intermediate d (1 mmol) was dissolved in 15 mL of methanol, and lithium hydroxide (10 mmol) was added to react at 70°C for 2 h. After TLC monitoring (petroleum ether: ethyl acetate = 2:1), the reaction solution was spin-dried, water was added to ultrasonically disperse the solid, and formic acid was added dropwise under stirring at room temperature. The pH was adjusted to 5-7, and the solid precipitated and filtered to obtain the hydrolyzed product e;

[0041] (6) The hydrolysis product e (1 mmol) obtained in the previous step was dissolved in 15 mL of dichloromethane, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.2 mmol) were added. The mixture was stirred at room temperature for 0.5 h, and O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (1.5 mmol) was added. The reaction was continued for 3 h. After TLC monitoring, the mixture was extracted with dichloromethane and water three times. The lower organic phases were combined, washed with saturated brine, and subjected to column chromatography (eluents: ethyl acetate and petroleum ether) to obtain intermediate f.

[0042] (7) Intermediate f was dissolved in dichloromethane, and trifluoroacetic acid was slowly added dropwise under stirring to remove the protecting group. The precipitated solid was filtered, and the filter cake after filtration was ultrasonically dispersed in water. The mixture was adjusted to weak acidity with saturated sodium bicarbonate solution, and filtered to obtain the 1,2,4-triazole compound (Compound Y-1) of this Example shown in Formula I. The yield of Compound Y-1 of this Example was 85%, and the compound was a white solid with a melting point of 232.6-233.2°C. The NMR characterization results were: 1 H NMR (400MHz, DMSO-d6) δ10.62(s,1H),8.09–8.03(m,2H),7.76–7.68(m,4H),7.60(t,J=7.6Hz,4H),7.55–7.45(m,4H),4.11(s,2H). 13C NMR(101MHz,DMSO-d6)δ166.50,164.27,160.95,152.01,141.71,137.61,131.03,13 0.07,129.64,129.37,128.29,126.41,125.42,119.14,35.50.HR-MS(ESI),calcd.C 23 H 19 N5O3,[MH]-m / z:412.1410,found:412.1414.

[0043] Example 2

[0044] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-2, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with o-methylbenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-2 in this example is 87%, and it is an off-white solid with a melting point of 230.6-232.1°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.57(s,1H),8.95(d,J=3.7Hz,1H),7.98(d,J=7.2Hz,1 H),7.76–7.69(m,4H),7.63–7.51(m,5H),7.32(q,J=7.1,5.8Hz,3H),4.12(s,2H),2.63(s,3H). 13 C NMR(101MHz,DMSO-d6)δ168.63,166.67,163.88,153.12,143.83,139.84,139.06,133.93,1 32.18,131.59,130.44,130.36,128.53,127.36,121.26,37.64,24.40.HR-MS(ESI),calcd.C 24 H 21 N5O3,[MH]-m / z:426.1566,found:426.1570.

[0045] Example 3

[0046] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-3, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with m-methylbenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-3 in this example is 82%, and it is a white solid with a melting point of 233.6-235.5°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),10.71(s,1H),10.28(s,1H),7.89(s,2H),7.72(dd,J=8. 5,4.3Hz,4H),7.60(q,J=8.4,7.9Hz,5H),7.37(s,1H),7.27(s,1H),4.11(s,2H),2.38(s,3H). 13 C NMR(101MHz,DMSO-d6)δ166.51,161.02,151.90,141.73,138.55,137.61,130.95,130.61,130.06,1 29.61,129.27,128.30,128.18,126.95,125.39,123.59,119.12,35.48,22.26.HR-MS(ESI),calcd.C 24 H 21 N5O3,[MH]-m / z:426.1566,found:426.1570.

[0047] Example 4

[0048] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-4, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methylbenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-4 in this example is 85%, and it is a white solid with a melting point of 233.9-235.2°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.12(d,J=6.6Hz,1H),10.66(s,1H),8.95(s,1H),7.96–7.92(m,2H),7.71(m,J=6. 6,2.2Hz,4H),7.60(q,J=7.6Hz,4H),7.53(d,J=7.1Hz,1H),7.30(d,J=7.9Hz,2H),4.10(s,2H),2.36(s,3H). 13C NMR(101MHz,DMSO-d6)δ166.51,164.52,161.14,151.82,141.74,139.47,137.63,130.0 5,129.94,129.58,128.32,126.37,125.39,119.11,35.26,21.49.HR-MS(ESI),calcd.C 24 H 21 N5O3,[MH]-m / z:426.1566,found:426.1570.

[0049] Example 5

[0050] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-5, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-ethylbenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. Compound Y-5 of this example has a yield of 79%, is a white solid, and has a melting point of 230.9-232.4°C. The NMR characterization results are: 1 H NMR(400MHz,DMSO-d6)δ11.13(s,1H),8.97(s,1H),7.98–7.95(m,2H),7.73–7.69(m,4H),7.63–7.5 7(m,4H),7.54–7.51(m,1H),7.33(d,J=8.0Hz,2H),4.10(s,2H),2.66(d,J=7.6Hz,2H),1.22(s,3H). 13 C NMR (101MHz, DMSO-d6) δ166.55,164.27,161.03,151.83,145.68,141.76,137.63,130.06,129. 57,128.74,128.56,128.31,126.46,125.37,119.09,35.50,28.55,15.86.HR-MS(ESI),calcd.C 25 H 23 N5O3,[MH]-m / z:440.1723,found:440.1727.

[0051] Example 6

[0052] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-6, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-isopropylbenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-6 in this example is 84%, and it is a white solid with a melting point of 232.9-235.1°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.57(s,1H),7.97(d,J=8.0Hz,2H),7.71(dd,J=8.2,3.6Hz,4H),7 .59(dd,J=8.3,6.4Hz,5H),7.36(d,J=8.0Hz,2H),4.09(s,2H),2.98–2.90(m,1H),1.24(d,J=6.8Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ166.49,161.04,151.81,150.28,141.69,137.66,130.05,129.55,1 28.72,128.31,127.26,126.49,125.38,119.13,35.53,33.82,24.25.HR-MS(ESI),calcd.C 26 H 25 N5O3,[MH]-m / z:454.1879,found:454.1884.

[0053] Example 7

[0054] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-7, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-7 in this example is 81%, and it is a white solid with a melting point of 231.7-234.0°C. The NMR characterization results are: 1 H NMR(400MHz,DMSO-d6)δ11.10(s,1H),10.72–10.52(m,1H),8.01–7.95(m,2H),7.7 9–7.45(m,9H),7.04(d,J=8.7Hz,2H),4.07(d,J=2.0Hz,2H),3.82(d,J=3.8Hz,3H). 13CNMR(101MHz,DMSO-d6)δ166.64,160.98,160.71,151.67,141.64,137.66,130.04,129. 56,128.31,127.91,125.37,123.62,119.11,114.74,56.98,36.22.HR-MS(ESI),calcd.C 24 H 21 N5O4,[MH]-m / z:442.1516,found:442.1520.

[0055] Example 8

[0056] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-8, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with 3,4,5-methoxybenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-8 in this example is 71%, and it is a white solid with a melting point of 237.9-240.1°C. The NMR characterization results are: 1 H NMR(400MHz,DMSO-d6)δ11.09(s,1H),10.55(s,1H),8.93(s,1H),7.74–7.68(m,4H ),7.61–7.51(m,5H),7.32(d,J=1.4Hz,2H),4.09(s,2H),3.85(s,6H),3.72(s,3H). 13 C NMR(101MHz,DMSO-d6)δ166.52,164.36,160.82,153.73,151.91,141.66,139.01,13 7.55,130.07,128.33,126.55,125.60,125.16,119.10,103.47,61.41,56.41,35.32.

[0057] Example 9

[0058] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-9, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-chlorobenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-9 in this example is 85%, and it is a white solid with a melting point of 232.9-234.5°C. The nuclear magnetic resonance characterization results are: 1H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 10.57 (s, 1H), 8.96 (s, 1H), 8.06 (d, J = 8.2 Hz, 2H), 7.71 (d, J = 8.0 Hz, 4H), 7.58 (dq, J = 19.5, 6.9, 5.5 Hz, 7H), 4.10 (s, 2H). 13 C NMR(101MHz,DMSO-d6)δ167.12,164.63,160.32,153.25,138.04,135.37,130.09,12 9.87,129.74,129.52,128.30,128.16,125.45,119.13,35.48.HR-MS(ESI),calcd.C 23 H 18 ClN5O3,[MH]-m / z:446.1020,found:446.1023.

[0059] Example 10

[0060] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-10, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with o-chlorobenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-10 in this example is 82%, and it is a white solid with a melting point of 233.9-235.7°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ10.72(s,1H),8.03(s,2H),7.74–7.68(m,4H),7.60(dd,J=7.7,5.2,2.6Hz,4H),7.56–7.51(m,3H),4.12(d,J=2.9Hz,2H). 13C NMR(101MHz,DMSO-d6)δ166.52,159.63,152.45,141.63,137.53,134.16,133.03,131.5 1,130.10,129.82,128.27,125.91,125.45,124.74,119.14,35.26.HR-MS(ESI),calcd.C 23 H 18 ClN5O3,[MH]-m / z:446.1020,found:446.1023.

[0061] Example 11

[0062] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-11, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-fluorobenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-11 in this example is 85%, and it is a white solid with a melting point of 234.1-236.8°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),10.58(d,J=2.7Hz,1H),8.97(s,1H),8.12–8. 06(m,2H),7.75–7.69(m,4H),7.62–7.51(m,5H),7.33(t,J=8.8Hz,2H),4.10(s,2H). 13 CNMR(101MHz,DMSO-d6)δ166.41,162.77,160.19,152.11,141.66,137.55,130.07,129.6 8,128.68,128.59,128.31,125.44,119.14,116.48,116.26,35.51.HR-MS(ESI),calcd.C 23 H 18 FN5O3,[MH]-m / z:430.1316,found:430.1320.

[0063] Example 12

[0064] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-12, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-bromobenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-12 in this example is 75%, and it is a white solid with a melting point of 233.4-235.8°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ10.69(d,J=13.6Hz,1H),8.01–7.97(m,2H),7.73–7.68(m,6H),7.62–7.57(m,4H),7.54(d,J=7.2Hz,1H),4.14–4.08(m,2H). 13 C NMR(101MHz,DMSO-d6)δ168.60,162.30,154.50,143.88,139.71,134.62, 132.45,132.28,131.92,130.62,130.45,127.63,125.49,121.33,38.07.

[0065] Example 13

[0066] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-13, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-nitrobenzaldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-13 in this example is 68%, and it is a yellow solid with a melting point of 232.4-236.5°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.15–11.07(m,1H),10.57(d,J=2.9Hz,1H),8.94(s,1H),8.38–8.29(m,2H ),8.20(s,1H),8.05(d,J=8.4Hz,1H),7.76–7.69(m,4H),7.64–7.54(m,5H),4.13(d,J=7.8Hz,2H). 13 C NMR(101MHz,DMSO-d6)δ169.96,166.48,164.14,159.73,152.76,148.56,141.59,13 7.56,130.14,128.31,127.48,125.52,124.84,119.14,35.54.HR-MS(ESI),calcd.C 23 H 18N6O5,[MH]-m / z:457.1262,found:457.1265.

[0067] Example 14

[0068] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-14, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with 3-thiophenealdehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-14 in this example is 83%, and it is a white solid with a melting point of 229.2-231.7°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ10.68 (s, 1H), 8.07 (dd, J = 3.0, 1.3Hz, 1H), 7.74–7.48 (m, 12H), 4.08 (s, 2H). 13 C NMR(101MHz,DMSO-d6)δ166.48,158.25,151.68,141.70,137.55,133.06,130.04,12 9.58,128.27,127.92,126.42,125.46,124.83,119.14,35.39.HR-MS(ESI),calcd.C 21 H 17 N5O3S,[MH]-m / z:418.0974,found:418.0978.

[0069] Example 15

[0070] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-15, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with cyclopropyl aldehyde, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly, and the 1,2,4-triazole compound (Compound Y-15) of this Example is finally prepared. The yield of Compound Y-15 of this Example is 82%, and it is a white solid with a melting point of 228.7-232.6°C. The NMR characterization results are: 1H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.50(s,1H),9.03–8.88(m,1H),7.71(d,J=8.4Hz,2H),7.62 –7.52(m,6H),7.46(t,J=7.3Hz,1H),3.94(s,2H),2.02(s,1H),0.90(ddt,J=37.4,5.2,2.7Hz,4H). 13 C NMR (101MHz, DMSO-d6) δ167.49,164.90,150.73,141.68,137.68,129.93,129.19,128.28,125.14,119.08,35.36,9.13,7.27.HR-MS(ESI),calcd.C 20 H 19 N5O3,[MH]-m / z:376.1410,found:376.1414.

[0071] Example 16

[0072] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-16, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with m-methylphenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-16 in this example is 84%, and it is a white solid with a melting point of 230.7-233.6°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.13(s,1H),10.61(s,1H),8.96(s,1H),8.07–8.03(m,2H),7. 72(d,J=8.5Hz,2H),7.61(d,J=8.3Hz,2H),7.54–7.40(m,7H),4.10(s,2H),2.38(s,3H). 13 C NMR(101MHz,DMSO-d6)δ166.49,160.87,151.96,141.76,139.79,137.53,131.07,130.21,1 29.82,129.35,128.28,126.40,125.85,122.38,119.10,35.26,21.36.HR-MS(ESI),calcd.C 24 H 21 N5O3,[MH]-m / z:426.1566,found:426.1570.

[0073] Example 17

[0074] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-17, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with p-methylphenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-17 in this example is 81%, and it is a white solid with a melting point of 231.1-233.4°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.58(s,1H),8.95(s,1H),8.06–8.02(m,2H),7.72(d,J=8 .7Hz,2H),7.62–7.56(m,4H),7.51–7.44(m,3H),7.39(d,J=8.1Hz,2H),4.06(s,2H),2.38(s,3H). 13 C NMR(101MHz,DMSO-d6)δ166.47,164.50,160.85,151.92,141.69,139.34,135.19,131.11,1 30.45,129.90,129.34,128.24,126.39,125.34,119.15,36.00,20.12.HR-MS(ESI),calcd.C 24 H 21 N5O3,[MH]-m / z:426.1566,found:426.1570.

[0075] Example 18

[0076] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-18, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with p-isopropylphenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-18 in this example is 84%, and it is a white solid with a melting point of 231.5-234.1°C. The NMR characterization results are: 1H NMR (400MHz, DMSO-d6) δ10.81(s,1H),8.08–8.01(m,2H),7.71(d,J=7.8Hz,2H),7.61(d, J=8.3Hz,4H),7.51–7.42(m,5H),4.10(s,2H),3.01–2.93(m,1H),1.22(d,J=6.9Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ166.09,159.36,157.96,152.13,149.10,145.11,141.86,136.02,1 29.00,127.38,126.37,125.39,120.36,119.05,35.78,33.42,24.22.HR-MS(ESI),calcd.C 26 H 25 N5O3,[MH]-m / z:454.1879,found:454.1883.

[0077] Example 19

[0078] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-19, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with p-iso-tert-butylphenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-19 in this example is 79%, and it is a white solid with a melting point of 233.5-236.2°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),10.55(s,1H),8.95(s,1H),8.08–8.02(m,2H),7.72(d,J=8. 4Hz, 2H), 7.60 (t, J = 6.5Hz, 6H), 7.47 (dd, J = 12.0, 7.1Hz, 3H), 4.08 (s, 2H), 1.31 (d, J = 1.3Hz, 9H). 13 C NMR(101MHz,DMSO-d6)δ166.52,164.36,160.83,152.22,151.92,141.69,135.11,131.0 7,128.30,128.17,126.84,126.29,125.10,119.13,35.06,31.35.HR-MS(ESI),calcd.C 27 H 27N5O3,[MH]-m / z:468.2036,found:468.2040.

[0079] Example 20

[0080] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-20, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with p-methoxyphenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-20 in this example is 83%, and it is a white solid with a melting point of 231.4-233.3°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.55(s,1H),8.95(s,1H),8.04(d,J=7.4Hz,2H),7.72(d,J= 8.5Hz,2H),7.63–7.58(m,4H),7.51–7.43(m,3H),7.12(d,J=8.9Hz,2H),4.03(s,2H),3.81(s,3H). 13 C NMR(101MHz,DMSO-d6)δ166.54,164.37,160.68,160.10,152.00,141.99,131.14,130.2 6,129.19,128.17,127.23,126.21,119.50,116.22,56.02,36.22.HR-MS(ESI),calcd.C 24 H 21 N5O4,[MH]-m / z:442.1516,found:442.1520.

[0081] Example 21

[0082] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-21, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with 3,4-methylphenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-21 in this example is 83%, and it is a white solid with a melting point of 230.5-233.5°C. The NMR characterization results are: 1H NMR (400MHz, DMSO-d6) δ11.13(s,1H),10.63(s,1H),8.95(s,1H),8.06–8.02(m,2H),7. 72(d,J=8.5Hz,2H),7.61(d,J=8.4Hz,2H),7.52–7.30(m,7H),4.07(s,2H),2.27(s,6H). 13 C NMR (101MHz, DMSO-d6) δ166.51,164.38,160.76,151.88,141.75,138.28,138.07,135.33,131.15,130. 74,129.86,129.33,128.28,126.38,126.30,122.63,119.11,35.48,19.85,19.56.HR-MS(ESI),calcd.C 25 H 23 N5O3,[MH]-m / z:440.1723,found:440.1727.

[0083] Example 22

[0084] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-22, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with 2,4-methylphenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-22 in this example is 80%, and it is a white solid with a melting point of 233.1-237.5°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),10.56(s,1H),8.95(s,1H),8.07–8.03(m,2H),7.72(d,J=8.4H z,2H),7.60(d,J=8.4Hz,2H),7.51–7.44(m,3H),7.29(s,2H),7.14(s,1H),4.08(s,2H),2.33(s,6H). 13 C NMR(101MHz,DMSO-d6)δ166.53,164.38,160.79,151.89,141.73,139.89,137.43,131.0 7,128.32,128.18,126.38,123.35,122.53,119.10,35.60,21.29.HR-MS(ESI),calcd.C 25 H23 N5O3,[MH]-m / z:440.1723,found:440.1727.

[0085] Example 23

[0086] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-23, having the structural formula: The difference between the preparation process and Example 1 is that the phenylhydrazine in step (1) is replaced with p-fluorophenylhydrazine, and the other steps are the same as Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-23 in this example is 84%, white solid, melting point: 230.6-233.4℃. The NMR characterization results are: 1 HNMR(400MHz,DMSO-d6)δ11.12(s,1H),10.63(s,1H),8.08–8.03(m,2H),7.80–7.75(m,2 H),7.74–7.70(m,2H),7.60(dd,J=8.8,1.9Hz,2H),7.51–7.42(m,5H),4.12–4.07(m,2H). 13 CNMR(101MHz,DMSO-d6)δ166.45,163.82,160.96,160.42,152.23,141.66,133.97,130.98,1 30.00,129.37,128.30,128.02,126.42,119.14,117.06,116.83,35.39.HR-MS(ESI),calcd.C 23 H 18 FN5O3,[MH]-m / z:430.1316,found:430.1320.

[0087] Example 24

[0088] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-24, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with m-fluorophenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-24 in this example is 81%, and it is a white solid with a melting point of 219.6-232.5°C. The NMR characterization results are: 1H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.55(s,1H),8.94(s,1H),8.05(d,J=7.2Hz,2H),7 .78–7.68(m,3H),7.63(d,J=6.6Hz,1H),7.49(ddt,J=27.4,18.3,7.7Hz,7H),4.02(s,2H). 13 C NMR(101MHz,DMSO-d6)δ165.83,161.51,158.04,153.49,141.64,132.77,130.79,13 0.13,129.58,129.42,128.28,126.43,125.87,119.04,34.96.HR-MS(ESI),calcd.C 23 H 18 FN5O3,[MH]-m / z:430.1316,found:430.1320.

[0089] Example 25

[0090] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-25, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with o-fluorophenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-25 in this example is 83%, and it is a white solid with a melting point of 231.6-233.9°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.60(s,1H),8.94(s,1H),8.08–8.03(m,2H),7.73–7.58(m,7H),7.52–7.39(m,4H),4.16(s,2H). 13 C NMR(101MHz,DMSO-d6)δ166.50,164.33,161.10,152.26,131.95,130.79,130.12,12 9.41,128.33,126.47,121.54,119.11,116.68,112.99,35.78.HR-MS(ESI),calcd.C 23 H 18 FN5O3,[MH]-m / z:430.1316,found:430.1320.

[0091] Example 26

[0092] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-26, having the structural formula: The difference between the preparation process and Example 1 is that the phenylhydrazine in step (1) is replaced with m-chlorophenylhydrazine, and the other steps are the same as Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-26 in this example is 84%, and it is a white solid with a melting point of 231.4-234.6°C. The NMR characterization results are: 1 H NMR(400MHz,DMSO-d6)δ11.12(s,1H),10.67(d,J=4.0Hz,1H),8.95(s,1H),8.08–8.04(m,2H),7.8 5(d,J=1.9Hz,1H),7.74–7.70(m,3H),7.63–7.58(m,4H),7.49(dd,J=7.8,2.3Hz,3H),4.16(s,2H). 13 C NMR(101MHz,DMSO-d6)δ166.46,164.27,161.15,152.30,141.64,138.79,134.21,131.76,13 0.77,130.14,129.41,128.31,126.48,125.26,124.09,119.14,35.26.HR-MS(ESI),calcd.C 23 H 18 ClN5O3,[MH]-m / z:446.1020,found:446.1024.

[0093] Example 27

[0094] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-27, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with p-chlorophenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-27 in this example is 81%, and it is a white solid with a melting point of 235.7-237.9°C. The NMR characterization results are: 1 H NMR(400MHz,DMSO-d6)δ11.13(s,1H),10.61(d,J=2.3Hz,1H),8.07–8.03(m,2H),7.78– 7.71(m,4H),7.67(d,J=8.8Hz,2H),7.62–7.58(m,2H),7.52–7.45(m,3H),4.12(s,2H). 13C NMR(101MHz,DMSO-d6)δ166.42,165.04,161.13,152.23,141.62,136.49,134.13,13 0.88,130.10,129.38,128.31,127.21,126.45,119.17,35.48.HR-MS(ESI),calcd.C 23 H 18 ClN5O3,[MH]-m / z:446.1020,found:446.1024.

[0095] Example 28

[0096] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-28, having the structural formula: The preparation process differs from that of Example 1 in that the phenylhydrazine in step (1) is replaced with p-bromophenylhydrazine, and the other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-28 in this example is 78%, and it is a light yellow solid with a melting point of 226.7-229.8°C. The NMR characterization results are: 1 H NMR(400MHz,DMSO-d6)δ11.14(s,1H),10.75(d,J=9.1Hz,1H),8.97(s,1H),8.07–8.04(m,2H),7 .80(d,J=8.4Hz,2H),7.74–7.69(m,4H),7.62(d,J=8.5Hz,2H),7.52–7.46(m,3H),4.15(s,2H). 13 C NMR(101MHz,DMSO-d6)δ166.45,164.44,161.15,152.23,141.70,136.90,133.03,130.8 7,130.07,129.38,128.30,127.42,126.46,122.59,119.16,35.49.HR-MS(ESI),calcd.C 23 H 18 BrN5O3,[MH]-m / z:490.0515,found:490.0519.

[0097] Example 29

[0098] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-29, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with p-methylphenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-29 in this example is 80%, and it is a white solid with a melting point of 225.1-228.0°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ10.53(s,1H),7.97(dd,J=9.0,2.0Hz,2H),7.70(d,J=7.6Hz,2H),7.56(d,J=7 .9Hz,4H),7.37(d,J=7.8Hz,2H),7.03(d,J=8.5Hz,2H),4.09–3.98(m,2H),3.81(s,3H),2.37(s,3H). 13 C NMR (101MHz, DMSO-D6) δ166.48,160.74,160.68,151.68,141.19,139.18,135.22 ,130.42,128.01,127.87,125.27,123.66,119.04,114.71,55.71,35.48,21.20.

[0099] Example 30

[0100] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-30, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-anisaldehyde and the phenylhydrazine is replaced with o-methylphenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-30 in this example is 78%, and it is a white solid with a melting point of 216.5-218.2°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.09(s,1H),10.43(d,J=7.2Hz,1H),8.93(s,1H),8.00–7.94(m,2H),7.69(dd,J=8.8,2.3Hz,2H),7.54( dd,J=8.6,3.1Hz,2H),7.50–7.43(m,3H),7.39–7.34(m,1H),7.06–7.01(m,2H),3.87(d,J=2.4Hz,2H),3.81(s,3H),2.14(s,3H). 13C NMR(101MHz,DMSO-D6)δ166.17,160.90,160.67,152.44,141.65,136.40,136 .19,128.29,128.13,127.85,123.80,119.08,115.33,114.61,35.04,18.05.

[0101] Example 31

[0102] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-31, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with m-methylphenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-31 in this example is 76%, and it is a white solid with a melting point of 264.9-265.6°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.57(s,1H),8.94(s,1H),7.98(d,J=8.5Hz,2H),7.72(dd,J=8.8,1.9Hz,2 H),7.63–7.58(m,2H),7.50–7.43(m,3H),7.33(s,1H),7.06–7.02(m,2H),4.07(s,2H),3.81(s,3H),2.37(s,3H). 13 C NMR (101MHz, DMSO-D6) δ166.56,160.79,160.72,151.65,130.10,129.80,128.31 ,128.17,127.90,125.81,123.60,122.32,119.08,114.72,56.24,35.78,21.75.

[0103] Example 32

[0104] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-32, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with p-isopropylphenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-32 in this example is 77%, and it is a white solid with a melting point of 275.9-277.4°C. The NMR characterization results are: 1H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.55(s,1H),8.95(s,1H),7.99–7.94(m,2H),7.74–7.69(m,2H),7.59(dd,J=8.4, 2.8Hz,4H),7.44(d,J=8.1Hz,2H),7.06–7.02(m,2H),4.04(s,2H),3.81(s,3H),3.00–2.93(m,1H),1.22(d,J=6.9Hz,6H). 13 C NMR (101MHz, DMSO-D6) δ166.56,160.75,151.63,149.88,141.70,135.45,128.28,12 8.16,127.86,127.83,125.36,123.66,119.11,114.72,55.71,35.46,33.67,24.24.

[0105] Example 33

[0106] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-33, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with 2,4-methylphenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-33 in this example is 76%, and it is a white solid with a melting point of 268.4-269.3°C. The NMR characterization results are as follows: 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),10.24(s,1H),10.04(s,1H),7.99–7.95(m,2H),7.72(d,J=8.3Hz,2H),7 .61(d,J=8.4Hz,2H),7.27(s,2H),7.13(s,1H),7.04(d,J=8.5Hz,2H),4.07(s,2H),3.81(s,3H),2.32(s,6H). 13 C NMR (101MHz, DMSO-D6) δ165.86,161.44,160.17,157.72,155.19,153.65,141.62 ,131.89,129.71,128.28,127.96,124.72,123.35,118.87,54.40,35.04,14.73.

[0107] Example 34

[0108] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-34, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with 3,4-methylphenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-34 in this example is 68%, and it is a white solid with a melting point of 266.8-267.7°C. The NMR characterization results are as follows: 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),10.56(s,1H),8.95(s,1H),8.07–8.03(m,2H),7.72(d,J=8.4H z,2H),7.60(d,J=8.4Hz,2H),7.51–7.44(m,3H),7.29(s,2H),7.14(s,1H),4.08(s,2H),2.33(s,6H). 13 C NMR (101MHz, DMSO-D6) δ166.60,164.15,160.67,151.57,142.14,141.74,141.56 ,138.25,135.37,128.32,127.53,123.65,119.09,115.75,51.07,35.53,19.46.

[0109] Example 35

[0110] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-35, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with p-methoxyphenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-35 in this example is 76%, and it is a white solid with a melting point of 271.5-272.0°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ10.54(s,1H),7.97(dd,J=8.8,1.6Hz,2H),7.71(dd,J=8.6,2.8Hz,2H), 7.58(td,J=6.9,3.4Hz,4H),7.10(d,J=8.9Hz,2H),7.05–7.01(m,2H),4.00(s,2H),3.81(s,6H). 13C NMR(101MHz,DMSO-D6)δ166.53,164.06,160.65,160.01,151.75,141.31,130.56 ,128.94,128.06,127.84,124.77,118.91,115.34,114.27,57.20,55.13,35.26.

[0111] Example 36

[0112] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-36, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with p-chlorophenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-36 in this example is 72%, and it is a white solid with a melting point of 264.1-264.3°C. The NMR characterization results are: 1 H NMR(400MHz,DMSO-d6)δ11.12(s,1H),10.61(s,1H),8.95(s,1H),8.01–7.96(m,2H),7.84(s,1H) ,7.75–7.68(m,3H),7.60(dt,J=6.0,2.7Hz,4H),7.05(d,J=8.4Hz,2H),4.13(s,2H),3.82(s,3H). 13 C NMR (101MHz, DMSO-D6) δ166.50,161.09,160.85,152.02,141.62,138.87,134.19,13 1.73,129.45,128.33,128.00,125.18,124.01,119.13,114.76,55.74,35.54,35.04.

[0113] Example 37

[0114] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-37, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with o-chlorophenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-37 in this example is 77%, and it is a white solid with a melting point of 267.8-268.6°C. The NMR characterization results are: 1H NMR (400MHz, DMSO-d6) δ11.12(s,1H),10.61(s,1H),8.95(s,1H),7.99(d,J=8.2Hz,2H),7.84(d,J=2.1H z,1H),7.72(dt,J=7.0,2.3Hz,3H),7.63–7.57(m,4H),7.05(d,J=8.8Hz,2H),4.13(s,2H),3.81(s,3H). 13 C NMR (101MHz, DMSO-D6) δ166.50,161.09,160.85,152.01,141.63,138.86,134.19,131 .72,129.45,128.33,128.00,125.19,124.01,123.32,119.14,114.76,55.73,35.53.

[0115] Example 38

[0116] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-38, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with p-bromophenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-38 in this example is 68%, and it is a white solid with a melting point of 266.6-267.7°C. The NMR characterization results are as follows: 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.58(s,1H),8.95(s,1H),7.97(d,J=8.4Hz,2H),7.81–7. 77(m,2H),7.73–7.66(m,4H),7.59(d,J=8.3Hz,2H),7.06–7.02(m,2H),4.09(s,2H),3.81(s,3H). 13 C NMR (101MHz, DMSO-D6) δ166.51,161.05,160.82,151.92,136.94,133.01,128.30,127.96,127.35,123.40,122.45,119.13,114.77,55.36,36.22.

[0117] Example 39

[0118] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-39, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with o-fluorophenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-39 in this example is 70%, and it is a white solid with a melting point of 261.6-262.5°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.10(s,1H),10.48(s,1H),8.94(s,1H),7.97(d,J=8.5Hz,2H),7.76–7.67(m,3H),7. 62(d,J=5.9Hz,1H),7.56–7.50(m,3H),7.41(d,J=7.6Hz,1H),7.05(d,J=8.8Hz,2H),3.98(s,2H),3.82(s,3H). 13 C NMR (101MHz, DMSO-D6) δ166.58,160.70,151.60,141.76,139.42,137.48,131.06,128.14,124.34,122.99,122.36,119.01,115.19,56.02,36.22.

[0119] Example 40

[0120] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-40, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with o-fluorophenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-40 in this example is 74%, and it is a white solid with a melting point of 265.4-266.3°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.10(s,1H),10.59(s,1H),8.93(s,1H),7.99(d,J=8.6Hz,2H),7.74–7. 70(m,2H),7.66–7.58(m,5H),7.38(d,J=2.6Hz,1H),7.07–7.03(m,2H),4.14(s,2H),3.82(s,3H). 13C NMR(101MHz,DMSO-D6)δ167.05,163.74,160.84,151.98,141.64,139.04,131 .91,128.33,127.99,123.33,121.46,119.11,114.76,112.85,55.73,35.56.

[0121] Example 41

[0122] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-41, having the structural formula: The preparation process differs from that of Example 1 in that the benzaldehyde in step (1) is replaced with p-methoxybenzaldehyde and the phenylhydrazine is replaced with p-fluorophenylhydrazine. The other steps are the same as those of Example 1. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-41 in this example is 72%, and it is a white solid with a melting point of 270.8-271.2°C. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.09(s,1H),10.57(s,1H),8.96(s,1H),7.98(d,J=8.5Hz,2H),7.78–7. 70(m,4H),7.63–7.58(m,2H),7.43(t,J=8.7Hz,2H),7.07–7.02(m,2H),4.06(s,2H),3.81(s,3H). 13 C NMR (101MHz, DMSO-D6) δ166.53,160.87,160.76,151.94,141.65,134.09,129 .71,128.22,127.91,123.51,118.48,116.98,114.80,114.36,56.02,36.22.

[0123] Example 42

[0124] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-42, having the structural formula: The difference between the preparation process and Example 7 is that steps (6) and (7) are omitted, and the other steps are the same as Example 7. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-42 in this example is 86%, white solid, melting point: 237.7-240.4℃. The NMR characterization results are: 1H NMR (400MHz, DMSO-d6) δ10.66(s,1H),7.98(d,J=8.4Hz,2H),7.90(d,J=8.5Hz,1H),7.75 –7.48(m,7H),7.05(dd,J=8.9,2.1Hz,2H),4.04(d,J=43.1Hz,2H),3.82(d,J=2.0Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ170.10,167.41,166.72,160.74,151.65,143.20,137 .64,130.99,130.08,127.91,125.38,123.56,119.10,114.76,55.73,35.78.

[0125] Example 44

[0126] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-44, having the structural formula: The difference between the preparation process and Example 11 is that steps (6) and (7) are omitted, and the other steps are the same as Example 11. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-44 in this example is 83%, white solid, melting point: 245.6-248.8℃. The NMR characterization results are: 1 H NMR (400MHz, DMSO-d6) δ11.62(s,1H),10.95(s,1H),8.09(d,J=6.7Hz,2H),7.91(d,J=7 .7Hz,2H),7.72(d,J=9.7Hz,4H),7.59(d,J=7.7Hz,3H),7.36–7.29(m,2H),4.15(s,2H). 13 C NMR(101MHz,DMSO-d6)δ166.80,165.92,160.76,151.63,143.27,137.60, 130.07,129.42,127.93,125.50,124.80,124.00,119.18,114.99,35.26.

[0127] Example 46

[0128] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-46, having the structural formula: The difference between the preparation process and Example 23 is that steps (6) and (7) are omitted, and the other steps are the same as Example 23. Due to the change in raw materials, the structure of the product obtained in each step of the preparation also changes accordingly. The yield of compound Y-46 in this example is 87%, white solid, melting point: 239.3-242.0℃. The NMR characterization results are: 1 H NMR(400MHz,DMSO-d6)δ10.81(s,1H),8.07–8.03(m,2H),7.90(d,J=8.5Hz,2H),7 .77(dd,J=8.7,4.9Hz,2H),7.68(d,J=8.6Hz,2H),7.50–7.43(m,5H),4.12(s,2H). 13 C NMR(101MHz,DMSO-d6)δ166.73,165.86,160.11,152.04,143.26,137.46,130.11,128.69,127.52,125.48,119.16,116.27,35.51.HR-MS(ESI),calcd.C 23 H 19 FN6O2,[MH]-m / z:429.1476,found:429.1479.

[0129] Example 43

[0130] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-43, having the structural formula: The preparation route and preparation method of the compound are as follows:

[0131]

[0132] (1) p-Anisaldehyde (1.2 mmol) and phenylhydrazine (1.0 mmol) were placed in a 250 mL round-bottom flask, and 100 mL of 20% ethanol aqueous solution (ethanol: water = 2: 8, v / v) was added to dissolve the mixture. The mixture was reacted in an oil bath at 80°C for 6-8 h and monitored by TLC (petroleum ether: ethyl acetate = 20:1). After the reaction, the mixture was directly filtered while hot. The filter cake was rinsed with ethyl acetate in small amounts several times and then dried to obtain intermediate a'.

[0133] (2) The obtained intermediate a' (1 mmol) was dissolved in 100 mL of acetonitrile, and glycine methyl ester hydrochloride (1.5 mmol), TBHP (1.5 mmol), I2 (1.4 mmol), and triethylamine (0.2 mmol) were added to react in an oil bath at 90°C for 3 h. The mixture was monitored by TLC (petroleum ether: ethyl acetate = 10:1), extracted three times with ethyl acetate and water, and the upper organic phases were combined and washed with saturated brine. The organic phase was removed by rotation and subjected to column chromatography (eluents: ethyl acetate and petroleum ether) to obtain intermediate b';

[0134] (3) The intermediate b' (1 mmol) was dissolved in 25 mL of methanol, and lithium hydroxide (10 mmol) was added and hydrolyzed at 70 °C for 2 h. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 3:1). After the reaction, the reaction solution was dried to obtain the intermediate c'.

[0135] (4) Intermediate c' (1 mmol), methyl p-aminobenzoate (1.2 mmol), and 1-methylimidazole (2 mmol) were dissolved in 25 mL of acetonitrile and stirred for 5 min. N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (1.2 mmol) was added and reacted at room temperature for 3 h. After monitoring by TLC (petroleum ether:ethyl acetate = 3:1), the mixture was filtered to obtain intermediate d'.

[0136] (5) The dried intermediate d' (1 mmol) was dissolved in 15 mL of methanol, and lithium hydroxide (10 mmol) was added to react at 70°C for 2 h. After TLC monitoring (petroleum ether: ethyl acetate = 2:1), the reaction solution was spin-dried, and the solid was dispersed by ultrasonication at room temperature. Formic acid was added dropwise under stirring, and the pH was adjusted to 5-7. The solid precipitated and filtered to obtain the hydrolyzed product e';

[0137] (6) The hydrolysis product e' (1 mmol) was dissolved in 15 mL of dichloromethane, and Boc-hydrazine (1.2 mmol), N,N-diisopropylethylamine (2.5 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.5 mmol) were added and reacted for 4 h. The reaction was monitored by TLC (petroleum ether:ethyl acetate = 3:1). After the reaction was completed, most of the organic phase was removed by vortexing, and column chromatography (eluent: ethyl acetate and petroleum ether) was performed to obtain the intermediate f';

[0138] (7) After ultrasonically dispersing the intermediate f' in water, hydrochloric acid was added dropwise to remove the Boc group, and then filtered to obtain the 1,2,4-triazole compound of this embodiment (Compound Y-43) represented by Formula I'. The yield of Compound Y-43 was 82%, and the compound was a white solid with a melting point of 246.9-249.7°C. The NMR characterization results were: 1H NMR (400MHz, DMSO-d6) δ11.58(d,J=10.3Hz,1H),10.92(d,J=10.6Hz,1H),7.98(d,J=8.8Hz,2H),7.91(dd,J=8 .6,1.6Hz,2H),7.71(t,J=8.2Hz,4H),7.58(d,J=7.7Hz,3H),7.05(d,J=8.9Hz,2H),4.13(s,2H),3.81(s,3H). 13 C NMR(101MHz,DMSO-d6)δ167.49,160.25,151.90,143.48,136.46,130.94,130.16 ,130.04,128.59,125.41,125.19,125.06,119.08,116.50,116.22,68.14,33.86.

[0139] Example 45

[0140] This embodiment provides a 1,2,4-triazole compound, designated as compound Y-45, having the structural formula: The preparation process differs from Example 43 in that p-methoxybenzaldehyde is replaced with p-fluorobenzaldehyde. The other steps are the same as in Example 43. Due to the change in starting materials, the structure of the product obtained in each step of the preparation also changes accordingly. Compound Y-45 in this example has an 83% yield and is a white solid with a melting point of 245.6-248.8°C. The NMR characterization results are: 1 HNMR (400MHz, DMSO-d6) δ11.62(s,1H),10.95(s,1H),8.09(d,J=6.7Hz,2H),7.91(d,J=7 .7Hz,2H),7.72(d,J=9.7Hz,4H),7.59(d,J=7.7Hz,3H),7.36–7.29(m,2H),4.15(s,2H). 13 C NMR(101MHz,DMSO-d6)δ166.80,165.92,160.76,151.63,143.27,137.60, 130.07,129.42,127.93,125.50,124.80,124.00,119.18,114.99,35.26.

[0141] Example 47

[0142] The present invention provides a 1,2,4-triazole compound, designated as compound Y-47, having the structural formula: The preparation process differed from that of Example 43 in that phenylhydrazine was replaced with p-fluorophenylhydrazine. All other steps were identical to those of Example 43. Due to the change in starting materials, the structures of the products obtained in each step of the preparation also changed accordingly. Compound Y-47 in this example was obtained in an 81% yield as a white solid with a melting point of 247.4-249.9°C. The results of nuclear magnetic resonance characterization are as follows: 1H NMR (400 MHz, DMSO-d6) δ11.66 (s, 1H), 11.03 (s, 1H), 8.05 (dd, J = 8.1, 1.7 Hz, 2H), 7.92 (d, J = 8.5 Hz, 2H), 7.80–7.72 (m, 4H), 7.51–7.42 (m, 5H), 4.16 (s, 2H). 13C NMR (101 MHz, DMSO-d6) δ166.76, 165.89, 160.92, 152.17, 143.22, 133.92, 130.88, 130.07, 129.00, 127.92, 126.40, 125.52, 119.17, 116.92, 35.78.

[0143] Experimental Example 1: Determination of HDAC6 Selective Inhibitory Activity

[0144] The 1,2,4-triazole compounds (compounds Y-1 to Y-47) provided in Examples 1 to 47 of the present invention were assayed for in vitro enzyme activity, as follows: the test compound was prepared into a 10mM mother liquor with DMSO to ensure full dissolution, and the mother liquor was gradiently diluted with buffer to obtain a series of concentrations; fluorescent substrate Ac-Gly-Ala-Lys-AMC, recombinant human histone deacetylase-6 or recombinant human histone deacetylase-1 (HDAC6 / HDAC1) were sequentially added to a 96-well plate, and the volume was made up to 100 μL per well with reaction buffer after adding different concentrations of the test compound solution to ensure that the reaction system was consistent. The 96-well plate was placed in a shaking table and incubated at low speed for 15 minutes to ensure that the fluorescent substrate was completely hydrolyzed. The excitation wavelength was set to 360 nm and the emission wavelength was 460 nm using a microplate reader, and the fluorescence intensity in each well was measured and the data was recorded, while vorinostat (SAHA) was used as a control. The inhibition rate was calculated according to the following formula: Inhibition rate (%) = (positive control fluorescence value - compound fluorescence value) / (positive control fluorescence value - blank group fluorescence value) × 100%. The experimental data were fitted with a dose-response curve using nonlinear regression analysis, and the IC of the test compound against HDAC1 / HDAC6 was calculated based on the fitted curve. 50 The HDAC6 inhibitory activities of compounds Y-1 to Y-47 are shown in Table 1.

[0145] Table 1. Inhibitory activity of compounds Y-1 to Y-47 of the present invention against HDAC6

[0146]

[0147] As shown in Table 1, the 1,2,4-triazole compounds provided by the present invention have different degrees of inhibitory activity against HDAC6. Among them, compounds Y-1 to Y-17, Y-21 to Y-27, Y-29 to Y-31, Y-33 to Y-34, and Y-36 to Y-41 have better inhibitory activity against HDAC6 than the positive control SAHA. Furthermore, using SAHA as a positive control, some compounds were selected to determine their IC values for HDAC6 and HDAC1. 50 The selective activity of the compound on HDAC6 was investigated, and the test results are shown in Table 2.

[0148] Table 2. Inhibitory activity and selectivity of some compounds of the present invention against HDAC1 / HDAC6

[0149]

[0150] As shown in Table 2, the 1,2,4-triazole compounds provided by the present invention exhibited good selective inhibition of HDAC6 relative to the positive control SAHA. Compound Y-15 exhibited the best selective inhibition, followed by compounds Y-23, Y-17, Y-27, Y-16, Y-26, and Y-22.

[0151] Experimental Example 2: Tumor Cell Inhibitory Activity Test

[0152] The following HDAC6 high-expressing tumor cell lines were selected as test cells, including: AGS cells (human gastric adenocarcinoma cells), MKN-45 cells (human gastric cancer cells), and KYSE-410 cells (human esophageal squamous cell carcinoma cells). The cells in the logarithmic growth phase were adjusted to a density of 2500 cells per well with DMEM culture medium containing 10% fetal bovine serum, seeded in 96-well plates (100 μL per well), and pre-cultured in a 37°C, 5% CO2 incubator for 24 hours to allow the cells to adhere to the wall and enter a stable growth phase. The 1,2,4-triazole compound provided by the present invention was prepared and a gradient concentration was set (representative compounds are shown in Table 3), diluted with culture medium, 100 μL of drug-containing culture medium was added to each well, and 3 replicates were set for each concentration; a blank group of culture medium only, a solvent control group containing 0.1% DMSO, and a positive control group (SAHA) were set, and cultured in a CO2 incubator under the same conditions for 48 hours. Add 10 μL of MTT reagent and continue incubation for 4 hours. Discard the supernatant and add 200 μL of DMSO to each well. Shake on a shaker for 15 minutes. Finally, measure the absorbance at 490 nm using a microplate reader. According to the formula, inhibition rate % = (control group - drug group) / (control group - blank group) × 100%, GraphPadPrism 8.0 was used to fit and calculate IC50 The test results are shown in Table 3. Note: The error is less than 10%

[0153] Table 3. Evaluation of the in vitro antiproliferative activity of representative compounds on AGS, MKN-45, and KYSE-410 cells

[0154]

[0155] The results in Table 3 show that the 1,2,4-triazole compounds provided by the present invention have a good proliferation inhibitory effect on AGS cells. Among them, compounds Y-4, Y-7, Y-12, Y-15, Y-16, Y-26, and Y-27 have significantly better proliferation inhibitory effects on AGS cells than the positive control SAHA. In addition, compounds Y-4 and Y-26 have significantly better proliferation inhibitory effects on MKN-45 cells than the positive control SAHA.

[0156] Experimental Example 3: Test of the protective effect on myocardial cells

[0157] The frozen H9c2 cells (rat cardiomyocytes) were revived and then subcultured to the logarithmic growth phase according to the required ratio. After culture, they were digested with trypsin and resuspended in complete culture medium. The cell density was adjusted to 5×10 4 cells / mL, add 100 μL of cell suspension to each well of a 96-well plate (i.e., seed 5000 cells per well). Gently shake the 96-well plate to evenly distribute the cells and place in a 37°C, 5% CO2 incubator for 24 hours. After 24 hours, use an inverted microscope to observe the cell adhesion and growth status to ensure that the cell confluence is ≥80%. Prepare a 300 μM H2O2 solution with culture medium, discard the old culture medium in the 96-well plate, add 100 μL of 300 μM H2O2 solution to each well, place the 96-well plate in a 37°C, 5% CO2 incubator for 1.5 hours, and use the MTT method to detect cell survival rate to establish a cell damage model. When the H9c2 cell confluence was ≥80%, H9c2 cells were incubated with different concentrations of 1,2,4-triazole compounds of the present invention (5, 10, 20, 40, 80 μM) for 24 h to evaluate the toxicity of 1,2,4-triazole HDAC6 inhibitors on H9c2 cells. The optimal concentration of the screened compound was 10 μM.

[0158] In the specific experiment, when the H9c2 cell confluence was ≥80%, the H9c2 cells were incubated with the 1,2,4-triazole compound (10 μM) of the embodiment of the present invention for 24 hours, and the culture medium was discarded after incubation; new culture medium was added, and 300 μM H2O2 was treated for 1.5 hours to induce myocardial cell injury, and the protective effect of 1,2,4-triazole compounds on the myocardial cell injury model was evaluated. At the same time, LUT (luteolin, 10 μM) was used as a positive control. The above groupings all had three replicates. After incubation, 20 μL of MTT solution was added to each well, mixed gently, and the 96-well plate was placed in a 37°C, 5% CO2 incubator for 4 hours in the dark. After that, the supernatant of the 96-well plate was discarded (be careful to avoid touching the formazan crystals at the bottom of the wells), 150 μL of DMSO was added to each well, and the mixture was shaken in the dark at room temperature for 10 minutes to fully dissolve the formazan crystals. After that, the absorbance of each well was measured at 490 nm and the data was recorded to calculate the cell survival rate. The experimental data were expressed as the mean ± standard deviation (Mean ± SD) of three independent experiments. P < 0.05 was used as the criterion for determining significant differences. The results are shown in Table 4. "PF" represents the ratio of the survival rate of myocardial damaged cells in the presence / absence of compound pretreatment.

[0159] Table 4. Evaluation of the cardiomyocyte protective ability of some compounds

[0160]

[0161] The results in Table 4 show that the 1,2,4-triazole compounds of the present invention have different degrees of protective effects on cardiomyocytes, and the protective effects of most compounds are significantly better than those of the positive control LUT.

[0162] In summary, the present invention, through structural optimization and structure-activity relationship studies of 1,2,4-triazole compounds, explored highly selective HDAC6 inhibitory compounds and simultaneously conducted in vitro anti-tumor activity and cardiomyocyte protective capacity evaluation tests. These results confirm that 1,2,4-triazole compounds possess HDAC6-selective activity, as well as in vitro anti-tumor and cardiomyocyte protective activities. Therefore, the present invention not only enriches the skeleton types of such compounds in HDAC6 inhibitors but also provides new directions and data support for the development of anti-tumor drugs with low myocardial damage risk and selectivity.

Claims

1. A 1,2,4-triazole compound, characterized in that It has the structure shown in formula I: In formula I, R1 is H, -Ph-4-CH3, -Ph-3-CH3, -Ph-2-CH3, -Ph-4-C2H5, -Ph-4-C3H7, -Ph-4-OCH3, -Ph-3,4,5-OCH3, -Ph-4-Cl, -Ph-3-Cl, -Ph-4-F, -Ph-4-Br, -Ph-4-NO2, One of the following; R2 is one of H, 4-CH3, 3-CH3, 2-CH3, 4-C3H7, 4-C4H9, 4-OCH3, 3,4-CH3, 2,4-CH3, 4-F, 3-F, 3-F, 3-Cl, 4-Cl, and 4-Br; R3 is One of OH.

2. The 1,2,4-triazole compound according to claim 1, characterized in that The 1,2,4-triazole compound is selected from the following compounds, which are sequentially denoted as compounds Y-1 to Y-47:

3. A method for preparing a 1,2,4-triazole compound according to claim 1 or 2, characterized in that: The following technical routes are adopted: When R3 is OH, the preparation method of the 1,2,4-triazole compound comprises the following steps: Raw materials A and B are dissolved in solvent a, and heated under reflux to react to obtain intermediate compound C; intermediate compound C is dissolved in solvent b, and then reacted with raw material D, tert-butyl hydroperoxide, I2, and an alkaline substance, followed by column chromatography to obtain intermediate compound E; intermediate compound E is dissolved in solvent c, and then reacted with an alkaline substance to obtain intermediate compound F; intermediate compound F is dissolved in solvent d, and then reacted with raw material G and a condensation reagent to obtain intermediate compound H; intermediate compound H is dissolved in solvent e, and then reacted with an alkaline substance to obtain compound I, which is the 1,2,4-triazole compound; When R3 is When the 1,2,4-triazole compound is prepared, the method comprises the following steps: Raw materials A and B are dissolved in solvent a and subjected to heating reflux reaction to obtain intermediate compound C; intermediate compound C is dissolved in solvent b, and then reacted with raw material D, tert-butyl hydroperoxide, I2, and an alkaline substance, followed by column chromatography to obtain intermediate compound E; intermediate compound E is dissolved in solvent c, and then reacted with an alkaline substance to obtain intermediate compound F; intermediate compound F is dissolved in solvent d, and then reacted with raw material G and a condensation reagent to obtain intermediate compound H; intermediate compound H is dissolved in solvent e, and then reacted with an alkaline substance to obtain compound I; compound I is added to solvent f, and then reacted with O-(tetrahydro-2H-pyran-2-yl)hydroxylamine and a condensation reagent, followed by column chromatography to obtain intermediate compound J; intermediate compound J is dissolved in solvent g, and then reacted with an acidic substance, and then filtered to obtain compound K, which is the 1,2,4-triazole compound; When R3 is When the 1,2,4-triazole compound is prepared, the method comprises the following steps: The raw materials A and B are dissolved in solvent a and heated under reflux to react to obtain an intermediate compound C; the intermediate compound C is dissolved in solvent b, and then reacted with raw material D, tert-butyl hydroperoxide, I2, and an alkaline substance, followed by column chromatography to obtain an intermediate compound E; the intermediate compound E is dissolved in solvent c, and then reacted with an alkaline substance to obtain an intermediate compound F; the intermediate compound F is dissolved in solvent d, and then reacted with raw material G and a condensation reagent to obtain an intermediate compound H; the intermediate compound H is dissolved in solvent e, and then reacted with an alkaline substance to obtain compound I; compound I is dissolved in solvent h, and then reacted with Boc-hydrazine and a condensation agent to obtain an intermediate compound L; the intermediate compound L is dissolved in solvent i, and then reacted with an acidic substance, and then filtered to obtain compound M, which is the 1,2,4-triazole compound.

4. The method for preparing 1,2,4-triazole compounds according to claim 3, wherein The alkaline substance is triethylamine or lithium hydroxide.

5. The method for preparing 1,2,4-triazole compounds according to claim 3, wherein The condensation reagent is one or more of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate, 1-methylimidazole, N,N-diisopropylethylamine, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate.

6. The method for preparing 1,2,4-triazole compounds according to claim 3, characterized in that: The acidic substance is trifluoroacetic acid or hydrochloric acid.

7. The method for preparing 1,2,4-triazole compounds according to claim 3, characterized in that: The solvent is one or more of ethanol, acetonitrile, methanol, dichloromethane and water.

8. A use of the 1,2,4-triazole compound according to claim 1 or 2, characterized in that: The application is the use of 1,2,4-triazole compounds in the preparation of HDAC6 inhibitors and / or anti-gastric cancer drugs and / or myocardial protection drugs.

9. The use of the 1,2,4-triazole compound according to claim 8, characterized in that: The anti-gastric cancer drug is a drug that inhibits the proliferation activity of gastric cancer cells; the myocardial protection drug is a drug that enhances the activity of myocardial cells.

10. The use of the 1,2,4-triazole compound according to claim 9, characterized in that: The gastric cancer cells are AGS cells; the cardiomyocytes are H9c2 cells.

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