CN116283831B - P-nitrobenzene derivative and preparation method and application thereof - Google Patents

Abstract

The invention provides a p-nitrobenzene derivative, a preparation method and application thereof, and belongs to the technical field of medicines. The p-nitrobenzene derivative provided by the invention has novel structure and good anti-tumor activity, and can be used for preparing anti-tumor drugs.

Description

A p-nitrobenzene derivative and its preparation method and application

Technical Field

The present invention relates to the field of medical technology, and in particular to a p-nitrobenzene derivative and a preparation method and application thereof.

Background technique

Microtubules are mainly composed of heterodimers formed by α-tubulin and β-tubulin connected end to end. Microtubules continuously polymerize and depolymerize in cells, and are in a dynamic balance. The dynamic balance of microtubules is related to cell morphology, the formation of spindles in mitosis, and cell movement. Disruption of the dynamic balance of microtubules will block the cell cycle and lead to cell apoptosis. In recent decades, microtubules have been a key target for tumor drug therapy. Tubulin-targeted drugs such as paclitaxel, vincristine, and vinorelbine have been approved for clinical treatment of tumors. The colchicine site is one of the main targets of microtubules, which is located between the α and β subunits. Compared with other targets of microtubules, colchicine site inhibitors usually have anti-angiogenic activity and are not prone to multidrug resistance caused by overexpression of P-glycoprotein.

In recent years, some colchicine site inhibitors have been reported, among which colchicine is a tricyclic alkaloid isolated from plants, which can effectively inhibit tubulin polymerization and has broad-spectrum anti-tumor activity. Developing more types of colchicine site inhibitors to expand the types of anti-tumor drugs is a technical problem that needs to be solved at present.

Summary of the invention

The purpose of the present invention is to provide a p-nitrobenzene derivative and a preparation method and application thereof. The p-nitrobenzene derivative provided by the present invention has anti-tumor activity.

In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

The present invention provides a p-nitrobenzene derivative having a structure shown in Formula A or Formula B:

Wherein, the R ₁ is selected from -H, -OCH ₃ or -OH;

The R ₂ is selected from -H, -SCH ₃ , -OCH ₃ , -SO ₂ CH ₃ or -OH;

The R ₃ is selected from -H or -OCH ₃ ;

The _R4 is selected from

Said X is selected from -O- or -NH-;

Said n=0 or 1.

Preferably, the p-nitrobenzene derivative is any one of the following compounds:

The present invention provides a method for preparing a p-nitrobenzene derivative according to the above technical solution, comprising the following steps:

2-Fluoro-5-nitro-benzoic acid is subjected to a first substitution reaction with the raw material compound 1 to obtain the intermediate 1;

The intermediate 1 is subjected to a second substitution reaction with the raw material compound 2 to obtain a p-nitrobenzene derivative having a structure shown in formula A;

The structural formulas of the raw material compound 1, intermediate 1 and raw material compound 2 are shown below:

R ₄ -H;

Sulfonation reaction of p-fluoronitrobenzene with chlorosulfonic acid was performed to obtain intermediate 2;

The intermediate 2 is subjected to a third substitution reaction with the raw material compound 3 to obtain the intermediate 3;

The intermediate 3 is subjected to a fourth substitution reaction with thiomorpholine to obtain a p-nitrobenzene derivative having a structure shown in formula B;

The structural formulas of the intermediate 2, the raw material compound 3 and the intermediate 3 are shown below:

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , X and n are as defined in Formula A or Formula B.

Preferably, the first substitution reaction is carried out at room temperature, and the time of the first substitution reaction is 2 to 6 hours.

Preferably, the temperature of the second substitution reaction is 75-85° C. and the time is 2-6 hours.

Preferably, the sulfonation reaction is carried out at a temperature of 95 to 105° C. and for a time of 45 to 50 hours.

Preferably, the third substitution reaction is carried out at room temperature, and the time of the third substitution reaction is 2 to 6 hours.

Preferably, the temperature of the fourth substitution reaction is 75-85° C., and the time is 1.5-2.5 h.

The present invention provides the use of the p-nitrobenzene derivatives described in the above technical solution in the preparation of anti-tumor drugs.

Preferably, the tumor is lung cancer.

The present invention provides a p-nitrobenzene derivative. The p-nitrobenzene derivative provided by the present invention has a novel structure, has good anti-tumor activity, and can be used for the preparation of anti-tumor drugs.

Detailed ways

The present invention provides a p-nitrobenzene derivative having a structure shown in Formula A or Formula B:

Wherein, the R ₁ is selected from -H, -OCH ₃ or -OH;

The R ₂ is selected from -H, -SCH ₃ , -OCH ₃ , -SO ₂ CH ₃ or -OH;

The R ₃ is selected from -H or -OCH ₃ ;

The _R4 is selected from

Said X is selected from -O- or -NH-;

Said n=0 or 1.

In the present invention, the p-nitrobenzene derivative is preferably any one of the compounds in Table 1.

Table 1 Available types of p-nitrobenzene derivatives

The present invention provides a method for preparing the p-nitrobenzene derivatives described in the above technical solution, which is described in detail below. In the present invention, unless otherwise specified, the raw materials used are commercially available products well known to those skilled in the art.

In the present invention, the method for preparing a p-nitrobenzene derivative having a structure shown in formula A comprises the following steps:

2-Fluoro-5-nitro-benzoic acid is subjected to a first substitution reaction with the raw material compound 1 to obtain the intermediate 1;

The intermediate 1 is subjected to a second substitution reaction with the raw material compound 2 to obtain a p-nitrobenzene derivative having a structure shown in formula A;

The structural formulas of the raw material compound 1, intermediate 1 and raw material compound 2 are shown below:

Wherein, R ₁ , R ₂ , R ₃ , R ₄ and X are as defined in Formula A

The reaction formula for preparing a p-nitrobenzene derivative having a structure shown in formula A of the present invention is as follows:

The present invention conducts a first substitution reaction between 2-fluoro-5-nitro-benzoic acid and raw material compound 1 to obtain intermediate 1. In the present invention, the raw material compound 1 is preferably 4-methylthiobenzyl alcohol, 4-methoxybenzyl alcohol, 3,4-dimethoxybenzyl alcohol, 4-methylsulfonylbenzyl alcohol, vanillyl alcohol, 3,4,5-trimethoxybenzyl alcohol, 3-hydroxy-4-methoxybenzyl alcohol, 4-hydroxymethyl-2,6-dimethoxyphenol, 3,5-dimethoxybenzyl alcohol, 4-methylthiobenzylamine or 4-methoxybenzylamine. In the present invention, the molar ratio of 2-fluoro-5-nitro-benzoic acid to raw material compound 1 is preferably 1: (0.8-1.2), more preferably 1: 1. In the present invention, the first substitution reaction is preferably carried out in the presence of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, triethylamine and dichloromethane; wherein the molar ratio of the 2-fluoro-5-nitro-benzoic acid, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and triethylamine is preferably 1:(1.3-1.7):(4.5-5.5), more preferably 1:1.5:5; the dichloromethane is used as a reaction solvent in an amount sufficient to ensure the smooth progress of the first substitution reaction, and the present invention has no special limitation on this.

The present invention preferably adds 2-fluoro-5-nitrobenzoic acid, triethylamine and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate to dichloromethane, stirs at room temperature until the raw materials are completely dissolved and the solution is light yellow, and then adds 4-methylthiobenzyl alcohol to carry out the first substitution reaction. In the present invention, the first substitution reaction is preferably carried out at room temperature. In an embodiment of the present invention, the room temperature is specifically 25°C; the time of the first substitution reaction is preferably 2 to 6 hours, more preferably 3 hours. The present invention preferably monitors the reaction progress by TLC, and the developing solvent used is preferably petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is preferably (2 to 5): 1. After the first substitution reaction, the present invention preferably evaporates the solvent in the obtained product system under reduced pressure, adds ethyl acetate to the residue, and then washes with water, dries with anhydrous sodium sulfate and filters in sequence, concentrates the filtrate under reduced pressure, and separates and purifies the residue by silica gel column chromatography to obtain intermediate 1. In the present invention, the reagents used for separation and purification by silica gel column chromatography are preferably petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is preferably (3-6):1.

After obtaining the intermediate 1, the present invention conducts a second substitution reaction between the intermediate 1 and the raw material compound 2 to obtain a p-nitrobenzene derivative having a structure shown in formula A. In the present invention, the raw material compound 2 is preferably thiomorpholine, morpholine, tetrahydropyrrole, 1-acetylpiperazine, 1-piperazinecarboxylic acid ethyl ester, 1-(2-hydroxyethyl)piperazine, 4-piperazinylbenzonitrile, N-Boc-piperazine, 1-benzoylpiperazine, 1-amino-4-methylpiperazine, 1-(2-pyrimidinyl)piperazine, m-hydroxyphenylpiperazine, 1-(4-pyridinyl)piperazine or N-methylpiperazine. In the present invention, the molar ratio of the intermediate 1 to the raw material compound 2 is preferably 1: (0.8-1.2), more preferably 1: 1. In the present invention, the second substitution reaction is preferably carried out in the presence of potassium carbonate and N,N-dimethylformamide; wherein the molar ratio of the raw material compound 2 to potassium carbonate is preferably 1:(1.8-2.2), more preferably 1:2; the N,N-dimethylformamide is used as a reaction solvent in an amount sufficient to ensure the smooth progress of the second substitution reaction, and the present invention has no special limitation on this.

The present invention preferably adds intermediate 1, raw material compound 2 and potassium carbonate to N,N-dimethylformamide to carry out a second substitution reaction. In the present invention, the temperature of the second substitution reaction is preferably 75-85°C, more preferably 80°C; the time of the second substitution reaction is preferably 2-6h, more preferably 3h. The present invention preferably monitors the reaction progress by TLC, and the developing agents used are preferably petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to ethyl acetate is preferably (1-10): (1-2). After the second substitution reaction, the present invention preferably evaporates the solvent in the obtained product system under reduced pressure, adds ethyl acetate to the residue, and then washes with water, dries with anhydrous sodium sulfate and filters in sequence, concentrates the filtrate under reduced pressure, and separates and purifies the residue by silica gel column chromatography to obtain a p-nitrobenzene derivative having a structure shown in formula A. In the present invention, the reagents used for separation and purification by silica gel column chromatography are preferably petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to ethyl acetate is preferably (1-10): 1.

In the present invention, the preparation method of the p-nitrobenzene derivative having the structure shown in formula B comprises the following steps:

Sulfonation reaction of p-fluoronitrobenzene with chlorosulfonic acid was performed to obtain intermediate 2;

The intermediate 2 is subjected to a third substitution reaction with the raw material compound 3 to obtain the intermediate 3;

The intermediate 3 is subjected to a fourth substitution reaction with thiomorpholine to obtain a p-nitrobenzene derivative having a structure shown in formula B;

The structural formulas of the intermediate 2, the raw material compound 3 and the intermediate 3 are shown below:

Wherein, X and n are as defined in Formula B.

The reaction formula for preparing a p-nitrobenzene derivative having a structure shown in Formula B according to the present invention is as follows:

The present invention conducts a sulfonation reaction between p-fluoronitrobenzene and chlorosulfonic acid to obtain intermediate 2. In the present invention, the dosage ratio of p-fluoronitrobenzene to chlorosulfonic acid is preferably 28mmol: (7-9)mL, more preferably 28mmol: 8mL. The present invention preferably adds p-fluoronitrobenzene to chlorosulfonic acid under ice bath conditions, stirs for 5-10min, and then heats up to conduct a sulfonation reaction. In the present invention, the temperature of the sulfonation reaction is preferably 95-105°C, more preferably 100°C; the time of the sulfonation reaction is preferably 45-50h, more preferably 48h. The present invention preferably monitors the reaction progress by TLC, and the developing agent used is preferably petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is preferably 5:1. After the sulfonation reaction, the present invention preferably cools the obtained product system and mixes it with ice water, then extracts it with ethyl acetate, washes the organic phase with saturated sodium bicarbonate aqueous solution and saturated brine in turn, evaporates the organic phase under reduced pressure, and separates and purifies the residue by silica gel column chromatography to obtain intermediate 2. In the present invention, the reagents used for separation and purification by silica gel column chromatography are preferably petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is preferably 4:1.

After obtaining intermediate 2, the present invention conducts a third substitution reaction between the intermediate 2 and the raw material compound 3 to obtain intermediate 3. In the present invention, the raw material compound 3 is preferably 4-methylthiophenol, 4-aminoanisole or 4-methylthiobenzylamine; the molar ratio of the raw material compound 3 to the intermediate 2 is preferably 1: (0.8-0.9), and more preferably 1: 0.84. In the present invention, the third substitution reaction is preferably carried out in the presence of triethylamine and dichloromethane; wherein the molar ratio of the raw material compound 3 to triethylamine is preferably 1: (8-12), and more preferably 1: 10; the dichloromethane is used as a reaction solvent, and its amount can ensure that the third substitution reaction proceeds smoothly, and the present invention has no special limitation on this.

The present invention preferably adds intermediate 2, raw material compound 3 and triethylamine to dichloromethane to carry out a third substitution reaction. In the present invention, the third substitution reaction is preferably carried out at room temperature, and the time of the third substitution reaction is preferably 2 to 6 hours, more preferably 3 hours. The present invention preferably monitors the reaction progress by TLC, and the developing agent used is preferably petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is preferably 8:1. After the third substitution reaction, the solvent in the obtained product system is evaporated under reduced pressure, and the residue is separated and purified by silica gel column chromatography to obtain intermediate 3. In the present invention, the reagents used for separation and purification by silica gel column chromatography are preferably petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is preferably 15:1.

After obtaining intermediate 3, the present invention conducts a fourth substitution reaction between intermediate 3 and thiomorpholine to obtain a p-nitrobenzene derivative having a structure shown in formula B. In the present invention, the molar ratio of intermediate 3 to thiomorpholine is preferably 1: (2.8-3.2), more preferably 1: 3. In the present invention, the fourth substitution reaction is preferably carried out in the presence of potassium carbonate and N, N-dimethylformamide; wherein the molar ratio of intermediate 3 to potassium carbonate is preferably 1: (1.8-2.2), more preferably 1: 2; the N, N-dimethylformamide is used as a reaction solvent, and its amount can ensure that the fourth substitution reaction proceeds smoothly, and the present invention has no special limitation on this.

The present invention preferably adds intermediate 3, thiomorpholine and potassium carbonate to N,N-dimethylformamide to carry out the fourth substitution reaction. In the present invention, the temperature of the fourth substitution reaction is 75-85°C, more preferably 80°C; the time of the fourth substitution reaction is preferably 1.5-2.5h, more preferably 2h. The present invention preferably monitors the reaction progress by TLC, and the developing agent used is preferably petroleum ether and ethyl acetate, and the volume ratio of the petroleum ether to ethyl acetate is preferably 6:1. After the fourth substitution reaction, the present invention preferably evaporates the solvent in the obtained product system under reduced pressure, adds ethyl acetate to the residue, and then washes with water, dries with anhydrous sodium sulfate and filters in sequence, concentrates the filtrate under reduced pressure, and separates and purifies the residue by silica gel column chromatography to obtain a p-nitrobenzene derivative having a structure shown in formula B. In the present invention, the reagents used for separation and purification by silica gel column chromatography are preferably petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is preferably 8:1.

The present invention provides the use of the p-nitrobenzene derivatives described in the above technical solution in the preparation of anti-tumor drugs. In the present invention, the tumor is preferably lung cancer. In the present invention, the anti-tumor drug preferably includes an active ingredient and a pharmaceutically acceptable excipient, and the active ingredient is the p-nitrobenzene derivatives described in the above technical solution; the present invention has no special restrictions on the pharmaceutically acceptable excipients, and the pharmaceutically acceptable excipients of the types familiar to those skilled in the art can be used.

The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

Example 1

The synthesis steps of compound I-1 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 4-methylthiobenzyl alcohol (3.08g, 20mmol) and stir at room temperature for 10min. The mixture was reacted for 3 h under the conditions of HPLC (the developing solvent was petroleum ether:ethyl acetate = 4:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether:ethyl acetate = 3:1 by volume ratio) to obtain 5.73 g of a yellow solid, which was intermediate 1a, with a yield of 89.2%;

A 50 mL round-bottom flask was added with 10 mL of N, N-dimethylformamide (DMF), and then intermediate 1a (161 mg, 0.5 mmol), thiomorpholine (52 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 2:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 4:1) to obtain 184 mg of a yellow solid, which was compound I-1, with a yield of 91.2%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.49 (d, J=2.8Hz, 1H), 8.11 (dd, J=9.2, 2.8Hz, 1H), 7.33–7.28 (m, 2H), 7.20–7.16 (m ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.73, 156.79, 139.67, 139.49, 132.02, 129.46, 129.38, 128.23, 127.78, 126.54, 126.51, 121.34, 118.45, 66.99, 54.17, 27.15, 15.65. HR-MS ( ESI):calcd.for C ₁₉ H ₂₁ N ₂ O ₄ S ₂ , [M+H] ⁺ 405.09373; found: 405.09335.

Example 2

The synthesis steps of compound I-2 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), 1-acetylpiperazine (64 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 1:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 1:1 by volume ratio) to obtain 132 mg of a yellow solid, which was compound I-2, with a yield of 61.5%;

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.61 (d, J=2.8Hz, 1H), 8.21 (dd, J=9.1, 2.8Hz, 1H), 7.39 (dd, J=8.5, 2.1Hz, 2H), 7.28–7.25 (m, 2H), 6.96 (d, J=9.1Hz, 1H), 5.32 (s, 2H), 3.74–3.70 (m, 2H), 3.52–3.49 (m, 2H), 3.20–3.14 ( m, 4H), 2.50 (s, 3H), 2.11 (s, 3H). ¹³ CNMR (151MHz, CDCl ₃ )δ: 169.21, 165.64, 155.74, 139.63, 131.88, 129.46, 128.27, 127.90, 126.43, 126.42, 120.86, 117.66, 67.09, 51.58, 50.68, 45.59, 40.89, 21.29 ,15.54.HR-MS(ESI):calcd. for C ₂₁ H ₂₄ O ₅ N ₃ S, [M+H] ⁺ 430.14312; found: 430.14276.

Example 3

The synthesis steps of compound I-3 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), 1-piperazinecarboxylic acid ethyl ester (79 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol) were added, and the mixture was stirred in an 80 ° C oil bath for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 3: 1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 2: 1 by volume ratio) to obtain 161 mg of a yellow solid, which was compound I-3, with a yield of 70.0%.

¹ H NMR (600MHz, CDCl ₃ ) δ: 8.61 (d, J=2.8Hz, 1H), 8.22 (dd, J=9.2, 2.8Hz, 1H), 7.41–7.36 (m, 2H), 7.30–7.27 ( m, 2H), 6.96 (d, J=9.2Hz, 1H), 5.32 (s, 2H), 4.17 (q, J=7.1Hz, 2H), 3.57 (s, 4H), 3.16 (s, 4H), 2.50 (s, 3H), 1.29 (t, J=7.1Hz, 3H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.71, 156.01, 155.44, 139.85, 139.83, 139.58, 131.92, 129.38, 128.34, 127.90, 126.55, 120.88, 117.67, 67.04, 61.68, 51.17, 15.63, 14.6 6.HR-MS(ESI):calcd.for C ₂₂ H ₂₆ O ₆ N ₃ S, [M+H] ⁺ 460.15318; found: 460.15305.

Example 4

The synthesis steps of compound I-4 are as follows:

A 50 mL round-bottom flask was added with 10 mL of DMF, and then intermediate 1a (161 mg, 0.5 mmol), 1-(2-hydroxyethyl)piperazine (65 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was stirred in an oil bath at 80 °C for 3 h. The reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 1:1 by volume ratio). The solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 2:1 by volume ratio) to obtain 182 mg of a yellow solid, which was compound I-4, with a yield of 84.4%.

¹ H NMR (600MHz, CDCl ₃ ) δ: 8.57 (d, J=2.8Hz, 1H), 8.19 (dd, J=9.2, 2.8Hz, 1H), 7.42–7.36 (m, 2H), 7.28–7.25 ( m, 2H), 6.95 (d, J = 9.2Hz, 1H), 5.31 (s, 2H), 3.64 (t, J = 5.4Hz, 2H), 3.22–3.17 (m, 4H), 2.58 (d, J =5.3Hz, 2H), 2.56–2.54(m, 4H), 2.49(s, 3H). ¹³ CNMR (151MHz, CDCl ₃ )δ: 166.03, 155.89, 155.86, 139.46, 139.37, 131.98, 129.46, 128.35, 127.83, 126.48, 126.45, 120.65, 117.51, 66.97, 59.31, 57.85, 52.30, 51.4 1,15.59.HR-MS(ESI):calcd. for C ₂₁ H ₂₆ O ₅ N ₃ S, [M+H] ₊ 432.15877; found: 432.15771.

Example 5

The synthesis steps of compound I-5 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), 4-piperazinebenzonitrile (94 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 5:1 by volume ratio). The solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 5:1 by volume ratio) to obtain 194 mg of a yellow solid, which was compound I-5, with a yield of 79.4%.

¹ H NMR (600MHz, CDCl ₃ ) δ: 8.62 (d, J=2.8Hz, 1H), 8.24 (dd, J=9.2, 2.8Hz, 1H), 7.54–7.52 (m, 2H), 7.41–7.39 ( m, 2H), 7.27 (d, J=1.9Hz, 2H), 6.99 (d, J=9.2Hz, 1H), 6.83–6.81 (m, 2H), 5.33 (s, 2H), 3.37–3.34 (m , 4H), 3.33–3.30 (m, 4H), 2.48 (s, 3H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.86, 155.36, 152.78, 139.69, 133.59, 131.88, 129.67, 128.20, 127.85, 126.33, 120.73, 119.85, 117.10, 114.02, 100.78, 65.32, 50.54, 46 .41, 15.48.HR-MS(ESI):calcd. for C ₂₆ H ₂₅ O ₄ N4 _S , [M+H]+489.15910; found:489.15887.

Example 6

The synthesis steps of compound I-6 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), 1-benzoylpiperazine (95 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol) were added, and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 8:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 6:1 by volume ratio) to obtain 142 mg of a yellow solid, which was compound I-6, with a yield of 57.8%.

¹ H NMR (600MHz, CDCl ₃ ) δ: 8.64 (d, J=2.8Hz, 1H), 8.23 (ddd, J=9.2, 2.9, 0.7Hz, 1H), 7.47–7.44 (m, 2H), 7.43 ( dd, J＝2.9, 0.9Hz, 1H), 7.42–7.40 (m, 2H), 7.38–7.36 (m, 2H), 7.26–7.23 (m, 2H), 6.97 (d, J＝9.2Hz, 1H) , 5.30 (s, 2H), 3.89 (s, 2H), 3.54 (s, 2H), 3.19 (d, J=91.3Hz, 4H), 2.47 (s, 3H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 170.58, 165.56, 155.94, 140.14, 139.67, 135.27, 131.84, 130.08, 129.39, 128.64, 128.40, 127.99, 127.14, 126.48, 121.06, 117.94, 67.07, 1 5.60.HR-MS(ESI):calcd.for C ₂₆ H ₂₆ O ₅ N ₃ S, [M+H] ⁺ 492.15877; found:492.15823.

Example 7

The synthesis steps of compound I-7 are as follows:

Take a 50mL round-bottom flask, add 10mL DMF, then add intermediate 1a (161mg, 0.5mmol), 1-amino-4-methylpiperazine (58mg, 0.5mmol) and potassium carbonate (138mg, 1mmol), place in an 80℃ oil bath and stir for 3h, TLC detects the completion of the reaction (by volume ratio, the developing solvent is petroleum ether: ethyl acetate = 2: 1); evaporate the solvent in the obtained product system under reduced pressure, add 20mL ethyl acetate to the residue, wash twice with 20mL ultrapure water, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (by volume ratio, the reagent used is petroleum ether: ethyl acetate = 1: 1) to obtain 136mg of a yellow solid, which is compound I-7, with a yield of 65.2%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.99 (s, 1H), 8.83 (d, J=2.7Hz, 1H), 8.18 (dd, J=9.5, 2.7Hz, 1H), 7.43 (d, J=9.5 Hz, 1H), 7.40–7.34 (m, 2H), 7.30–7.27 (m, 2H), 5.29 (s, 2H), 2.98 (s, 4H), 2.70 (d, J=112.8Hz, 4H), 2.50 (s, 3H), 2.34 (s, 3H). ¹³ C NMR (151MHz, CDCl ₃ )δ:166.87, 154.09, 139.31, 136.76, 132.08, 129.82, 129.01, 128.88, 126.67, 112.64, 107.56, 66.60, 55.30, 54.67, 45.79, 15.70. HR-MS (ESI): calcd.for C ₂₀ H ₂₅ O ₄ N ₄ S, [M+H] ⁺ 417.15910; found: 417.15823.

Example 8

The synthesis steps of compound I-8 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), 1-(2-pyrimidinyl)piperazine (82 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 1:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 1:1) to obtain 157 mg of a yellow solid, which was compound I-8, with a yield of 67.5%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.60 (d, J=2.8Hz, 1H), 8.33 (d, J=4.7Hz, 2H), 8.20 (dd, J=9.3, 2.8Hz, 1H), 7.39 ( d, J=8.1Hz, 2H), 7.27 (d, J=8.2Hz, 2H), 6.98 (d, J=9.2Hz, 1H), 6.54 (t, J=4.7Hz, 1H), 5.33 (s, 2H), 3.97–3.89 (m, 4H), 3.29 (dd, J=6.2, 4.0Hz, 4H), 2.49 (s, 3H). ¹³ CNMR (151MHz, CDCl ₃ )δ: 165.96, 161.51, 157.77, 155.87, 139.36, 131.98, 129.37, 128.40, 127.82, 126.62, 120.36, 117.23, 110.42, 67.03, 50.98, 43.18, 15.64. HR-MS( ESI):calcd.for C ₂₃ H ₂₄ O ₄ N ₅ S, [M+H] ⁺ 466.15435; found: 466.15393.

Example 9

The synthesis steps of compound I-9 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), m-hydroxyphenylpiperazine (89 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol) were added, and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 1:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 1:1) to obtain 182 mg of a yellow solid, which was compound I-8, with a yield of 75.9%.

¹ HNMR (600 MHz, CDCl ₃ )δ: 8.58 (d, J = 2.8Hz, 1H), 8.22 (dd, J = 9.2, 2.8Hz, 1H), 7.46–7.41 (m, 2H), 7.29 (dq, J = 8.7, 2.2Hz, 2H), 7.13 (t, J = 8.1Hz, 1H), 6.98 (d, J = 9.2Hz, 1H), 6.50 –6.45 (m, 1H), 6.36 (ddd, J=8.0, 2.3, 0.7Hz, 1H), 6.27 (t, J=2.3Hz, 1H), 5.34 (s, 2H), 5.03 (s, 1H), 3.31–3.22 (m, 4H), 3.17–3.07 (m, 4H), 2.50 (s, 3H). ¹³ CNMR (151MHz, CDCl ₃ ) δ: 166.30, 156.73, 155.61, 152.18, 139.67, 139.28, 132.10, 130.19, 129.92, 128.08, 127.79, 126.48, 121.23, 117.25, 108. 63, 107.03, 102.91, 67.15, 51.14, 48.29, 15.49. HR-MS (ESI): calcd. for C ₂₅ H ₂₆ O ₅ N ₃ S, [M+H] ⁺ 480.15877; found: 480.15802.

Example 10

The synthesis steps of compound I-10 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), m-hydroxyphenylpiperazine (82 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol) were added, and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 5:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 4:1) to obtain 177 mg of a yellow solid, which was compound I-10, with a yield of 76.1%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.62 (d, J=2.8Hz, 1H), 8.39–8.29 (m, 2H), 8.24 (dd, J=9.2, 2.8Hz, 1H), 7.45–7.37 (m , 2H), 7.27 (d, J=2.0Hz, 2H), 6.98 (d, J=9.2Hz, 1H), 6.68–6.56 (m, 2H), 5.33 (s, 2H), 3.41–3.34 (m, 4H), 3.34–3.26 (m, 4H), 2.48 (s, 3H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.85, 155.40, 154.43, 150.42, 139.79, 139.72, 131.83, 129.69, 128.22, 126.29, 120.73, 117.09, 108.22, 67.18, 50.48, 45.25, 15.47.HR-MS( ESI):calcd.for C ₂₄ H ₂₅ O ₄ N ₄ S, [M+H] ⁺ 465.15910; found: 465.15814.

Embodiment 11

The synthesis steps of compound I-11 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), N-methylpiperazine (50 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol) were added, and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 1:2 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 1:1) to obtain 153 mg of a yellow solid, which was compound I-11, with a yield of 76.3%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.62 (t, J=2.9Hz, 1H), 8.22 (d, J=9.3Hz, 1H), 7.42–7.36 (m, 2H), 7.28 (s, 2H), 6.96 (d, J=9.3Hz, 1H), 5.32 (d, J=3.0Hz, 2H), 3.54 (s, 4H), 3.17 (s, 4H), 2.50 (d, J=3.1Hz, 3H), 1.27 (dt, J=10.1, 6.4Hz, 3H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.74, 156.06, 154.63, 139.73, 139.56, 131.93, 129.35, 128.45, 127.93, 126.55, 120.68, 117.58, 80.27, 67.01, 51.21, 28.41, 15.64. HR-MS (ES I):calcd.for C ₂₀ H ₂₄ O ₄ N ₃ S, [M+H] ⁺ 402.14820; found: 402.11099.

Example 12

The synthesis steps of compound I-12 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1a (161 mg, 0.5 mmol), tetrahydropyrrole (36 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 1:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 2:1) to obtain 144 mg of a yellow solid, which was compound I-12, with a yield of 77.4%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.50 (d, J=2.8Hz, 1H), 8.13 (dd, J=9.5, 2.8Hz, 1H), 7.41–7.37 (m, 2H), 7.29–7.26 (m ¹³ C NMR (151MHz, CDCl ₃ )δ: 166.68, 151.06, 139.27, 136.07, 132.14, 129.30, 128.44, 127.29, 126.54, 115.34, 113.38, 66.99, 51.34, 25.76, 15.70. HR-MS (ESI): calcd.for C ₁₉ H _{2 1} O ₄ N ₂ S, [M+H] ⁺ 373.12165; found: 373.12079.

Example 13

The synthesis steps of compound I-13 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 4-methoxybenzyl alcohol (2.76g, 20mmol) and stir at room temperature for 10min. The mixture was reacted for 3 h under the conditions of HPLC (the developing solvent was petroleum ether:ethyl acetate = 5:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether:ethyl acetate = 4:1 by volume ratio) to obtain 5.24 g of a yellow solid, which was intermediate 1b, with a yield of 85.9%;

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1b (153 mg, 0.5 mmol), thiomorpholine (52 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol) were added, and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 4:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 4:1) to obtain 174 mg of a yellow solid, which was compound I-13, with a yield of 89.7%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.55 (d, J=2.8Hz, 1H), 8.18 (dd, J=9.2, 2.8Hz, 1H), 7.42–7.39 (m, 2H), 6.96 (d, J =9.2Hz, 1H), 6.93–6.90 (m, 2H), 5.30 (s, 2H), 3.82 (s, 3H), 3.44–3.42 (m, 4H), 2.70–2.67 (m, 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.89, 159.95, 156.68, 139.68, 130.58, 128.17, 127.69, 127.53, 121.65, 118.35, 114.09, 67.18, 55.35, 54.13, 27.15. HR-MS (ESI): calcd.for C ₁₉ H _{2 1} O ₅ N ₂ S, [M+H] ⁺ 389.11657; found: 389.11636.

Embodiment 14

The synthesis steps of compound I-14 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1b (153 mg, 0.5 mmol), morpholine (44 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 4:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 4:1) to obtain 123 mg of a yellow solid, which was compound I-14, with a yield of 66.1%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.59 (d, J=2.8Hz, 1H), 8.21 (dd, J=9.2, 2.8Hz, 1H), 7.43–7.38 (m, 2H), 6.95 (d, J =9.2Hz, 1H), 6.93–6.90 (m, 2H), 5.30 (s, 2H), 3.82 (s, 3H), 3.76–3.72 (m, 4H), 3.17–3.13 (m, 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 166.05, 159.97, 155.93, 139.76, 130.68, 128.36, 127.87, 127.40, 121.11, 117.27, 114.07, 67.23, 66.33, 55.34, 51.68. HR-MS (ESI): calcd.for C ₁₉ H _{2 1} O ₆ N ₂ , [M+H] ⁺ 373.13941; found:373.13937.

Embodiment 15

The synthesis steps of compound I-15 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 3,4-dimethoxybenzyl alcohol (2.76g, 20mmol) and stir at room temperature. The reaction was carried out under the conditions of 4% paraformaldehyde and 1% paraformaldehyde. The reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 5:1 by volume); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 4:1 by volume) to obtain 6.17 g of a yellow solid, which was intermediate 1c, with a yield of 92.0%;

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1c (168 mg, 0.5 mmol), morpholine (44 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 6:1 by volume ratio). The solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 7:1) to obtain 156 mg of a yellow solid, which was compound I-15, with a yield of 77.4%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.65 (d, J=2.8Hz, 1H), 8.23 (dd, J=9.2, 2.8Hz, 1H), 6.97 (d, J=9.2Hz, 1H), 6.60 ( d, J＝2.3Hz, 2H), 6.44 (t, J＝2.3Hz, 1H), 5.30 (s, 2H), 3.81 (s, 6H), 3.79–3.75 (m, 4H), 3.22–3.15 (m , 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.77, 161.08, 156.10, 139.77, 137.49, 128.49, 128.00, 120.75, 117.39, 106.39, 100.41, 67.19, 66.34, 60.39, 55.42, 51.76, 21.05, 14.21. HR-MS(ESI):calcd.forC ₂₀ H ₂₃ O ₇ N ₂ , [M+H] ⁺ 403.14998; found: 403.14948.

Example 16

The synthesis steps of compound I-16 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1c (168 mg, 0.5 mmol), thiomorpholine (52 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 7:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 6:1) to obtain 135 mg of a yellow solid, which was compound I-16, with a yield of 69.7%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.50 (d, J=2.8Hz, 1H), 8.12 (dd, J=9.5, 2.8Hz, 1H), 7.03 (dd, J=8.1, 2.0Hz, 1H), 7.01 (d, J=2.1Hz, 1H), 6.88 (d, J=8.1Hz, 1H), 6.70 (d, J=9.4Hz, 1H), 5.30 (s, 2H), 3.92 (s, 3H), 3.89 (s, 3H), 3.32–3.26 (m, 4H), 1.99–1.93 (m, 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 166.75, 151.01, 149.10, 136.04, 128.44, 128.07, 127.23, 121.62, 115.48, 113.38, 111.97, 111.10, 67.37, 55.97, 51.33, 29.70, 25.76, 14.20 .HR-MS(ESI):calcd.for C ₂₀ H ₂₃ O ₆ N ₂ , [M+H] ⁺ 387.15506; found: 387.15445.

Embodiment 17

The synthesis steps of compound I-17 are as follows:

Take a 50mL round-bottom flask, add 10mL DMF, then add intermediate 1c (168mg, 0.5mmol), tetrahydropyrrole (36mg, 0.5mmol) and potassium carbonate (138mg, 1mmol), place in an 80℃ oil bath and stir for 3h, TLC detects the completion of the reaction (by volume ratio, the developing solvent is petroleum ether: ethyl acetate = 4:1); evaporate the solvent in the obtained product system under reduced pressure, add 20mL ethyl acetate to the residue, wash twice with 20mL ultrapure water, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (by volume ratio, the reagent used is petroleum ether: ethyl acetate = 6:1) to obtain 149mg of a yellow solid, which is compound I-17, with a yield of 71.1%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.59 (d, J=2.8Hz, 1H), 8.20 (dd, J=9.2, 2.8Hz, 1H), 7.04 (dd, J=8.1, 2.0Hz, 1H), 7.00 (d, J=2.0Hz, 1H), 6.97 (d, J=9.2Hz, 1H), 6.88 (d, J=8.2Hz, 1H), 5.31 (s, 2H), 3.91 (s, 3H), 3.90 (s, 3H), 3.47–3.42 (m, 4H), 2.73–2.68 (m, 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.82, 156.74, 149.12, 139.70, 128.27, 127.75, 121.78, 121.51, 118.41, 112.10, 111.13, 67.43, 60.39, 56.00, 54.15, 27.16, 21.05, 14.20. HR-MS(ESI):calcd.for C ₂₀ H ₂₃ O ₆ N ₂ S, [M+H] ⁺ 419.12713; found: 419.12692.

Embodiment 18

The synthesis steps of compound I-18 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 4-methylsulfonylbenzyl alcohol (3.72g, 20mmol) and stir at room temperature for 10min. The mixture was reacted for 3 h under the conditions of HPLC (the developing solvent was petroleum ether:ethyl acetate = 5:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether:ethyl acetate = 4:1 by volume ratio) to obtain 6.57 g of a yellow solid, which was intermediate 1d, with a yield of 92.9%;

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1d (177 mg, 0.5 mmol), thiomorpholine (52 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 2: 1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 2: 1) to obtain 177 mg of a yellow solid, which was compound I-18, with a yield of 80.9%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.81 (d, J=2.8Hz, 1H), 8.33 (dd, J=9.0, 2.8Hz, 1H), 8.03–7.98 (m, 2H), 7.66 (d, J =8.2Hz, 2H), 7.17 (d, J = 9.1Hz, 1H), 5.45 (s, 2H), 3.68 (dd, J = 7.1, 3.7Hz, 4H), 3.24 (t, J = 5.2Hz, 4H ), 3.09 (s, 3H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 171.14, 128.98, 128.61, 128.05, 120.79, 66.24, 60.39, 51.67, 51.17, 44.49, 21.05, 14.20. HR-MS (ESI): calcd.for C ₁₉ H ₂₁ O ₆ N ₂ S ₂ , [M +H] ⁺ 437.08355; found: 437.19324.

Embodiment 19

The synthesis steps of compound I-19 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1d (177 mg, 0.5 mmol), morpholine (44 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 2: 1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 1: 1) to obtain 151 mg of a yellow solid, which was compound I-19, with a yield of 71.7%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.69 (d, J=2.8Hz, 1H), 8.25 (dd, J=9.2, 2.8Hz, 1H), 8.01–7.97 (m, 2H), 7.66 (d, J =8.1Hz, 2H), 7.02 (d, J = 9.2Hz, 1H), 5.44 (s, 2H), 3.83–3.79 (m, 4H), 3.24–3.20 (m, 4H), 3.07 (s, 3H) . ¹³ C NMR (151MHz, CDCl ₃ )δ: 165.24, 156.49, 141.52, 140.72, 139.76, 128.95, 128.73, 128.38, 127.92, 119.68, 117.83, 66.35, 65.97, 51.90, 44.50. HR-MS (ESI): calcd.for C ₁₉ H _{2 1} O ₇ N ₂ S, [M+H] ⁺ 421.10640; found: 421.10605.

Embodiment 20

The synthesis steps of compound I-20 are as follows:

Take a 50mL round-bottom flask, add 10mL DMF, then add intermediate 1d (177mg, 0.5mmol), tetrahydropyrrole (36mg, 0.5mmol) and potassium carbonate (138mg, 1mmol), place in an 80℃ oil bath and stir for 3h, TLC detects the completion of the reaction (by volume ratio, the developing solvent is petroleum ether: ethyl acetate = 2: 1); evaporate the solvent in the obtained product system under reduced pressure, add 20mL ethyl acetate to the residue, wash twice with 20mL ultrapure water, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (by volume ratio, the reagent used is petroleum ether: ethyl acetate = 1: 1) to obtain 168mg of a yellow solid, which is compound I-20, with a yield of 82.9%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.56 (d, J=2.8Hz, 1H), 8.16 (dd, J=9.5, 2.8Hz, 1H), 8.01–7.97 (m, 2H), 7.67 (d, J =8.0Hz, 2H), 6.75 (d, J = 9.5Hz, 1H), 5.43 (s, 2H), 3.30 (q, J = 6.0, 4.7Hz, 4H), 3.08 (s, 3H), 2.01–1.97 (m, 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 166.09, 151.30, 141.75, 140.63, 136.13, 128.93, 128.62, 127.89, 127.57, 114.34, 113.66, 65.97, 51.53, 44.53, 25.77. HR-MS (ESI): calcd.for C ₁₉ H _{2 1} O ₆ N ₂ S, [M+H] ⁺ 405.11148; found: 405.11057.

Embodiment 21

The synthesis steps of compound I-21 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 4-hydroxy-3-methoxybenzyl alcohol (3.08g, 20mmol) and stir at room temperature. The mixture was reacted at room temperature for 3 h, and the reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 3:1 by volume); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 4:1 by volume) to obtain 6.21 g of a yellow solid, which was intermediate 1e, with a yield of 96.7%;

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1e (160 mg, 0.5 mmol), morpholine (44 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 4:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 6:1) to obtain 152 mg of a yellow solid, which was compound I-21, with a yield of 78.1%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.94 (d, J=2.8Hz, 1H), 8.29 (dd, J=9.2, 2.8Hz, 1H), 7.12 (d, J=8.0Hz, 1H), 7.09 ( d, J=1.9Hz, 1H), 7.03 (d, J=9.2Hz, 1H), 6.99 (dd, J=8.0, 1.9Hz, 1H), 4.72 (d, J=4.9Hz, 2H), 3.85 ( d, J=4.7Hz, 4H), 3.84 (s, 3H), 3.36 (dd, J=5.5, 3.7Hz, 4H), 1.90 (t, J=5.6Hz, 1H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 163.90, 156.64, 151.19, 140.54, 139.65, 138.64, 129.52, 128.56, 122.60, 119.14, 117.51, 111.01, 66.49, 64.94, 55.78, 51.83, 14.20. HR-MS (ES I):calcd.forC ₁₉ H ₂₁ O ₇ N ₂ , [M+H] ⁺ 389.13433; found: 389.13412.

Embodiment 22

The synthesis steps of compound I-22 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 3,4,5-trimethoxybenzyl alcohol (3.96g, 20mmol) and stir at room temperature. The mixture was reacted at room temperature for 3 h, and the reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 5:1 by volume); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 6:1 by volume) to obtain 6.88 g of a yellow solid, which was the intermediate 1f, with a yield of 94.2%;

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1f (183 mg, 0.5 mmol), morpholine (44 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 8:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 8:1) to obtain 177 mg of a yellow solid, which was compound I-22, with a yield of 77.8%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.66 (d, J=2.8Hz, 1H), 8.23 (dd, J=9.2, 2.8Hz, 1H), 6.97 (d, J=9.2Hz, 1H), 6.70 ( s, 2H), 5.29 (s, 2H), 4.64 (s, 3H), 3.85 (d, J=9.5Hz, 6H), 3.80–3.77 (m, 4H), 3.21–3.18 (m, 4H). ¹³ CNMR (151MHz, CDCl ₃ )δ: 165.74, 156.11, 153.46, 153.42, 139.74, 138.35, 137.40, 136.62, 130.85, 128.55, 128.05, 120.59, 117.46, 105.90, 103.86, 67.52, 66.35, 65 .56, 60.86, 56.11, 51.75.HR-MS (ESI ):calcd.for C ₂₁ H ₂₄ O ₈ N ₂ Na, [M+H] ⁺ 455.14249; found: 455.14166.

Embodiment 23

The synthesis steps of compound I-23 are as follows:

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1f (183 mg, 0.5 mmol), thiomorpholine (52 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 6:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 5:1) to obtain 175 mg of a yellow solid, which was compound I-23, with a yield of 77.9%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.63 (d, J=2.8Hz, 1H), 8.21 (dd, J=9.2, 2.8Hz, 1H), 6.99 (d, J=9.2Hz, 1H), 6.70 ( s, 2H), 5.29 (s, 2H), 3.89 (s, 6H), 3.86 (s, 3H), 3.49–3.45 (m, 4H), 2.73–2.70 (m, 4H). ¹³ CNMR (151MHz, CDCl ₃ )δ:165.64, 156.84, 153.46, 139.68, 138.32, 130.95, 128.36, 127.85, 121.21, 118.52, 105.85, 67.51, 60.89, 56.24, 54.18, 27.16. HR-MS (ESI): calcd.for C ₂₁ H ₂₅ O ₇ N ₂ S, [M+H] ⁺ 449.13770; found: 449.13663.

Embodiment 24

The synthesis steps of compound I-24 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 3-hydroxy-4-methoxybenzyl alcohol (3.08g, 20mmol) and stir at room temperature. The mixture was reacted for 3 h at room temperature, and the reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 4:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 4:1 by volume ratio) to obtain 6.04 g of a yellow solid, which was 1 g of an intermediate, with a yield of 94.0%;

Take a 50mL round-bottom flask, add 10mL DMF, then add intermediate 1g (160mg, 0.5mmol), thiomorpholine (52mg, 0.5mmol) and potassium carbonate (138mg, 1mmol), place in an 80℃ oil bath and stir for 3h, TLC detects the completion of the reaction (by volume ratio, the developing solvent is petroleum ether: ethyl acetate = 5:1); evaporate the solvent in the obtained product system under reduced pressure, add 20mL ethyl acetate to the residue, wash twice with 20mL ultrapure water, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (by volume ratio, the reagent used is petroleum ether: ethyl acetate = 4:1) to obtain 153mg of a yellow solid, which is compound I-24, with a yield of 75.5%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.85 (d, J=2.8Hz, 1H), 8.21 (dd, J=9.2, 2.8Hz, 1H), 7.21–7.19 (m, 1H), 7.13 (d, J =2.1Hz, 1H), 6.98 (d, J = 9.3Hz, 1H), 6.94 (d, J = 8.4Hz, 1H), 5.23 (s, 1H), 4.60 (s, 2H), 3.77 (s, 3H ), 3.57–3.55(m, 4H), 2.74–2.71(m, 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 163.70, 157.38, 150.60, 139.68, 139.40, 133.99, 129.38, 128.38, 125.91, 121.65, 119.74, 118.59, 112.40, 64.53, 55.98, 54.34, 27.30. HR-MS( ESI):calcd.for C ₁₉ H ₂₁ O ₆ N ₂ S, [M+H] ⁺ 405.11148; found: 405.11127.

Embodiment 25

The synthesis steps of compound I-25 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol), stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow, then add 4-hydroxymethyl-2,6-dimethoxyphenol (3.68g, 20mmol) , react for 3 hours at room temperature, and the reaction is completed by TLC detection (the developing solvent is petroleum ether: ethyl acetate = 4:1 by volume ratio); the solvent in the obtained product system is evaporated under reduced pressure, 50 mL of ethyl acetate is added to the residue, washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated under reduced pressure. The residue is separated and purified by silica gel column chromatography (the reagent used is petroleum ether: ethyl acetate = 5:1 by volume ratio) to obtain 6.21 g of yellow solid, which is intermediate 1h, with a yield of 88.4%;

A 50 mL round-bottom flask was added with 10 mL DMF, and then the intermediate 1h (176 mg, 0.5 mmol), 1-(tert-butyloxycarbonyl)piperazine (93 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol) were added, and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 8:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 8:1) to obtain 192 mg of a yellow solid, which was compound I-25, with a yield of 74.1%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.92 (d, J=2.8Hz, 1H), 8.26 (dd, J=9.2, 2.8Hz, 1H), 6.98 (d, J=9.2Hz, 1H), 4.71 ( s, 2H), 3.89 (s, 1H), 3.85 (s, 6H), 3.61–3.57 (m, 4H), 3.36 (s, 4H), 1.48 (s, 9H). ¹³ CNMR (151MHz, CDCl ₃ )δ: 164.07, 156.06, 154.72, 152.37, 140.11, 139.50, 129.18, 128.22, 127.22, 119.66, 117.20, 103.88, 103.16, 80.25, 65.34, 56.30, 56.13, 28.4 0.HR-MS(ESI):calcd.for C ₂₅ H ₃₂ O ₉ N ₃ , [M+H] ⁺ 518.21331; found: 518.21234.

Embodiment 26

The synthesis steps of compound I-26 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 3,5-dimethoxybenzyl alcohol (3.36g, 20mmol) and stir at room temperature. The reaction was carried out under the conditions of 40 ° C for 3 h, and the reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 6:1 by volume); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and it was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 6:1 by volume) to obtain 6.55 g of yellow solid, which was intermediate 1i, with a yield of 97.7%;

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1i (168 mg, 0.5 mmol), thiomorpholine (52 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was placed in an 80 ° C oil bath with stirring for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 10: 1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagents used were petroleum ether: ethyl acetate = 10: 1) to obtain 157 mg of a yellow solid, which was compound I-26, with a yield of 74.9%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.62 (d, J=2.8Hz, 1H), 8.21 (dd, J=9.2, 2.8Hz, 1H), 6.98 (d, J=9.2Hz, 1H), 6.59 ( d, J＝2.3Hz, 2H), 6.44 (t, J＝2.3Hz, 1H), 5.30 (s, 2H), 3.81 (s, 6H), 3.48–3.45 (m, 4H), 2.73–2.70 (m , 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ:165.64, 161.08, 156.85, 139.71, 137.61, 128.32, 127.82, 121.31, 118.45, 106.28, 100.41, 67.16, 55.44, 55.36, 54.21, 27.15. HR-MS (ESI): calcd.for C ₂₀ H ₂₃ O ₆ N ₂ S, [M+H] ⁺ 419.12713; found: 419.12653.

Embodiment 27

The synthesis steps of compound II-1 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 4-methylthiobenzylamine (3.06g, 20mmol) and stir at room temperature for 10min. The mixture was reacted for 3 h under the following conditions. The reaction was completed by TLC (the developing solvent was petroleum ether:ethyl acetate = 2:1 by volume ratio). The solvent in the obtained product system was evaporated under reduced pressure. 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether:ethyl acetate = 3:1 by volume ratio) to obtain 5.87 g of a yellow solid, which was intermediate 1j, with a yield of 91.6%.

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1j (160 mg, 0.5 mmol), tetrahydropyrrole (36 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was stirred in an 80 ° C oil bath for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 3:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagents used were petroleum ether: ethyl acetate = 4:1) to obtain 133 mg of a yellow solid, which was compound II-1, with a yield of 71.5%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.19 (d, J=2.7Hz, 1H), 8.07 (dd, J=9.4, 2.8Hz, 1H), 7.32–7.27 (m, 2H), 7.25 (d, J =8.3Hz, 2H), 6.61 (d, J = 9.4Hz, 1H), 6.29 (s, 1H), 4.56 (d, J = 5.8Hz, 2H), 3.41–3.32 (m, 4H), 2.49 (s , 3H), 2.01–1.92 (m, 4H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 168.51, 150.07, 134.39, 128.74, 126.97, 126.65, 126.25, 120.51, 113.33, 60.39, 50.41, 43.93, 25.65, 15.88, 14.20.HR-MS (ESI): calcd.for C ₁₉ H ₂₂ O _{3N 3} S, [M+H] ⁺ 372.13764; found: 372.13754.

Embodiment 28

The synthesis steps of compound II-2 are as follows:

A 50mL round-bottom flask was added with 10mL DMF, and then intermediate 1j (160mg, 0.5mmol), thiomorpholine (52mg, 0.5mmol) and potassium carbonate (138mg, 1mmol), and the mixture was placed in an 80°C oil bath with stirring for 3h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 4:1 by volume ratio). The solvent in the obtained product system was evaporated under reduced pressure, 20mL of ethyl acetate was added to the residue, washed twice with 20mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagents used were petroleum ether: ethyl acetate = 3:1 by volume ratio) to obtain 146mg of a yellow solid, which was compound II-2, with a yield of 72.3%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.91 (d, J=2.8Hz, 1H), 8.80 (s, 1H), 8.26 (dd, J=8.9, 2.8Hz, 1H), 7.33–7.29 (m, 2H ), 7.27(q, J＝4.2, 3.1Hz, 2H), 7.22(d, J＝8.8Hz, 1H), 4.62(d, J＝5.6Hz, 2H), 3.33–3.20(m, 4H), 2.61 –2.52 (m, 4H), 2.49 (s, 3H). ¹³ CNMR (151MHz, CDCl ₃ )δ: 164.43, 156.55, 143.89, 138.53, 134.61, 128.77, 127.40, 127.16, 126.87, 120.93, 55.14, 43.60, 27.63, 15.95. HR-MS (ESI): calcd.for C ₁₉ H ₂₂ O ₃ N ₃ S ₂ , [M+H] ⁺ 404.10971; found: 404.10925.

Embodiment 29

The synthesis steps of compound II-3 are as follows:

Take a 250mL round-bottom flask and add 50mL dichloromethane, 2-fluoro-5-nitrobenzoic acid (3.70g, 20mmol), triethylamine (10.10g, 100mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (11.40g, 30mmol). Stir at room temperature for 10min until the raw materials are completely dissolved and the solution is light yellow. Then add 4-methoxybenzylamine (2.74g, 20mmol) and stir at room temperature. The mixture was reacted for 3 h, and the reaction was completed by TLC (the developing solvent was petroleum ether:ethyl acetate = 2:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 50 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 50 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether:ethyl acetate = 3:1 by volume ratio) to obtain 5.14 g of a yellow solid, which was intermediate 1k, with a yield of 84.5%;

A 50 mL round-bottom flask was added with 10 mL DMF, and then intermediate 1k (152 mg, 0.5 mmol), thiomorpholine (52 mg, 0.5 mmol) and potassium carbonate (138 mg, 1 mmol), and the mixture was stirred in an oil bath at 80 ° C for 3 h. The reaction was completed by TLC detection (the developing solvent was petroleum ether: ethyl acetate = 3:1 by volume ratio); the solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, washed twice with 20 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (by volume ratio, the reagent used was petroleum ether: ethyl acetate = 2:1) to obtain 129 mg of a yellow solid, which was compound II-3, with a yield of 66.5%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.86 (d, J=2.8Hz, 1H), 8.75 (t, J=5.5Hz, 1H), 8.22 (dd, J=8.9, 2.9Hz, 1H), 7.35– 7.29 (m, 2H), 7.20 (d, J=8.9Hz, 1H), 6.95–6.88 (m, 2H), 4.59 (d, J=5.5Hz, 2H), 3.81 (s, 3H), 3.30–3.23 (m, 4H), 2.58–2.52 (m, 4H). ¹³ CNMR (151MHz, CDCl ₃ )δ: 164.35, 159.42, 156.55, 143.70, 129.88, 129.59, 128.82, 127.29, 126.74, 120.82, 114.41, 55.40, 55.07, 43.53, 27.54. HR-MS (ESI): calcd.for C ₁₉ H _{2 2} O ₄ N ₃ S, [M+H] ⁺ 388.13255; found: 388.13177.

Embodiment 30

The synthesis steps of compound III-1 are as follows:

Take a 25mL round-bottom flask, add 8mL chlorosulfonic acid, place it in an ice bath, add p-fluoronitrobenzene (4.00g, 28mmol), stir for 10min, then place the reaction solution in a 100℃ oil bath, stir and react for 48h, and detect the completion of the reaction by TLC (the developing solvent is petroleum ether: ethyl acetate = 5:1 by volume ratio); after cooling the obtained product system, add it dropwise to 300mL ice water, and then extract it twice with 300mL ethyl acetate, wash the organic phase with saturated sodium bicarbonate aqueous solution and saturated brine in turn, evaporate the organic phase under reduced pressure, and separate and purify the residue by silica gel column chromatography (the reagent used is petroleum ether: ethyl acetate = 4:1 by volume ratio) to obtain 2.74g of brown oil, which is intermediate 2, with a yield of 40.8%;

A 50 mL round-bottom flask was added with 10 mL of dichloromethane, intermediate 2 (200 mg, 0.84 mmol), 4-methylthioaniline (139 mg, 1 mmol) and triethylamine (1.01 g, 10 mmol), and the mixture was stirred at room temperature for 1 h. The reaction was completed by TLC (the developing solvent was petroleum ether:ethyl acetate = 8:1 by volume). The solvent in the obtained product system was evaporated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether:ethyl acetate = 15:1 by volume) to obtain 260 mg of a yellow oily product, which was intermediate 3a, with a yield of 90.5%.

A 50 mL round-bottom flask was added with 5 mL DMF, intermediate 3a (260 mg, 0.76 mmol), thiomorpholine (234 mg, 2.27 mmol) and potassium carbonate (209 mg, 1.51 mmol), and the mixture was stirred in an oil bath at 80 °C for 3 h. The reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 6:1 by volume). The solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 15 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagents used were petroleum ether: ethyl acetate = 8:1 by volume) to obtain 290 mg of a yellow solid, which was compound III-1, with a yield of 89.6%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.76 (d, J=2.7Hz, 1H), 8.39 (dd, J=8.8, 2.7Hz, 1H), 7.56 (s, 1H), 7.47 (d, J=8.7 Hz, 1H), 7.11–7.08 (m, 2H), 6.92–6.90 (m, 2H), 3.43 (dd, J＝6.4, 3.6Hz, 4H), 2.98 (dd, J＝5.7, 4.0Hz, 4H) , 2.42 (s, 3H). ¹³ C NMR (151MHz, CDCl ₃ )δ: 157.11, 144.32, 136.57, 135.69, 132.97, 129.04, 127.59, 126.78, 124.33, 122.12, 56.50, 28.49, 15.86. HR-MS (ESI): calcd.for C ₁₇ H ₂₀ O ₄ N ₃ S ₃ , [M+H] ⁺ 426.06104; found: 426.06042.

Embodiment 31

The synthesis steps of compound III-2 are as follows:

A 50 mL round-bottom flask was added with 10 mL of dichloromethane, intermediate 2 (200 mg, 0.84 mmol), 4-methylthiophenol (140 mg, 1 mmol) and triethylamine (1.01 g, 10 mmol), and the mixture was stirred at room temperature for 1 h. The reaction was completed by TLC (the developing solvent was petroleum ether:ethyl acetate = 8:1 by volume). The solvent in the obtained product system was evaporated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether:ethyl acetate = 15:1 by volume) to obtain 244 mg of a yellow oily product, which was intermediate 3b, with a yield of 84.7%.

A 50 mL round-bottom flask was added with 5 mL DMF, intermediate 3b (261 mg, 0.76 mmol), thiomorpholine (234 mg, 2.27 mmol) and potassium carbonate (209 mg, 1.51 mmol), and the mixture was stirred in an oil bath at 80 °C for 3 h. The reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 6:1 by volume). The solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 15 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether: ethyl acetate = 8:1 by volume) to obtain 255 mg of a yellow solid, which was compound III-2, with a yield of 78.7%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.70 (d, J=2.7Hz, 1H), 8.34 (dd, J=9.0, 2.7Hz, 1H), 7.27 (d, J=9.0Hz, 1H), 7.11– 7.09(m, 2H), 6.89–6.87(m, 2H), 3.42–3.40(m, 4H), 2.79–2.76(m, 4H), 2.38(s, 3H). ¹³ CNMR (151MHz, CDCl ₃ )δ:158.47，146.74，142.34，138.45，130.56，129.87，128.67，127.64，123.72，122.19，55.81，27.65，15.92.HR-MS(ESI):calcd.forC ₁₇ H ₁₉ O ₅ N ₂ S ₃ ，[ M+H] ⁺ 427.04506; found: 427.04468.

Embodiment 32

The synthesis steps of compound III-3 are as follows:

A 50 mL round-bottom flask was added with 10 mL of dichloromethane, intermediate 2 (200 mg, 0.84 mmol), 4-methylthiobenzylamine (153 mg, 1 mmol) and triethylamine (1.01 g, 10 mmol), and the mixture was stirred at room temperature for 1 h. The reaction was completed by TLC (the developing solvent was petroleum ether:ethyl acetate = 8:1 by volume). The solvent in the obtained product system was evaporated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (the reagent used was petroleum ether:ethyl acetate = 15:1 by volume) to obtain 234 mg of a yellow oily product, which was intermediate 3c, with a yield of 78.3%.

A 50 mL round-bottom flask was added with 5 mL DMF, intermediate 3c (270 mg, 0.76 mmol), thiomorpholine (234 mg, 2.27 mmol) and potassium carbonate (209 mg, 1.51 mmol), and the mixture was placed in an 80 °C oil bath with stirring for 3 h. The reaction was completed by TLC (the developing solvent was petroleum ether: ethyl acetate = 6:1 by volume). The solvent in the obtained product system was evaporated under reduced pressure, 20 mL of ethyl acetate was added to the residue, and the mixture was washed twice with 15 mL of ultrapure water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (the reagents used were petroleum ether: ethyl acetate = 8:1 by volume) to obtain 271 mg of a yellow solid, which was compound III-3, with a yield of 81.1%.

¹ HNMR (600MHz, CDCl ₃ ) δ: 8.76 (d, J=2.7Hz, 1H), 8.30 (dd, J=8.7, 2.7Hz, 1H), 7.21 (d, J=8.8Hz, 1H), 6.96– 6.93(m, 2H), 6.83–6.79(m, 2H), 5.85(t, J＝5.9Hz, 1H), 3.98(d, J＝5.7Hz, 2H), 3.05(dd, J＝6.2, 3.7Hz , 4H), 2.64–2.61 (m, 4H), 2.35 (s, 3H). ¹³ CNMR (151MHz, CDCl ₃ )δ: 156.86, 144.26, 139.29, 137.34, 132.06, 128.66, 128.36, 126.41, 126.09, 124.34, 55.99, 47.91, 28.30, 15.54. HR-MS (ESI): calcd.for C ₁₈ H ₂₂ O ₄ N ₃ S ₃ , [M+H] ⁺ 440.07669; found: 440.07642.

Test Example 1

The p-nitrobenzene derivatives prepared in Examples 1 to 32 were subjected to an in vitro antitumor activity test (CCK8 test), as follows:

A549 tumor cells in the logarithmic growth phase were collected, and a cell suspension with a concentration of 40,000 cells/mL was adjusted with an automatic cell counter, added to a 96-well plate at 100 μL/well, and incubated for 24 hours to adhere to the wall. The compounds to be tested were diluted with culture medium into drug solutions with concentrations of 5 μM and 1 μM, respectively, and the culture medium containing the drug solution was added after the culture medium in the 96-well plate was discarded. A blank control group, a negative control group, a positive control group (IMB5046), and a drug-adding group were set up, with three replicates in each group, and incubated in an incubator for 48 hours after drug addition. After discarding the original culture medium, 100 μL of new culture medium and 10 μL of CCK8 reagent were added to each well, and incubated in the dark for 1 hour. The absorbance value was measured at a wavelength of 450 nm, and the inhibition rate of the proliferation of A549 tumor cells by the compounds to be tested at different concentrations was calculated according to the following formula, and the specific results are shown in Table 2.

Table 2 Activity test results of the compounds prepared in Examples 1 to 32

The above pharmacological experimental data show that the p-nitrobenzene derivatives provided by the present invention have high anti-tumor cell activity, especially compounds I-1, I-2, I-12, I-13 and I-14 have good anti-tumor activity. The p-nitrobenzene derivatives provided by the present invention can be further studied and developed as drug lead molecules to improve their drugability.

The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims (5)

1. A p-nitrobenzene derivative characterized by the following formula:

。

2. the method for producing p-nitrobenzene derivatives as defined in claim 1, comprising the steps of:

carrying out a first substitution reaction on 2-fluoro-5-nitro-benzoic acid and 4-methylthiobenzyl alcohol to obtain an intermediate 1, wherein the intermediate 1 has the following formula structure:

；

And carrying out a second substitution reaction on the intermediate 1 and 1-acetyl piperazine to obtain the p-nitrobenzene derivative.

3. The preparation method according to claim 2, wherein the first substitution reaction is performed at room temperature for 2 to 6 hours.

4. The method according to claim 2, wherein the temperature of the second substitution reaction is 75-85 ℃, and the time of the second substitution reaction is 2-6 hours.

5. The use of the p-nitrobenzene derivative as defined in claim 1 for preparing anti-tumor drugs, wherein the tumor is lung cancer.