Benzofurazan derivatives, processes for their preparation and use

Abstract

This invention discloses benzofuran derivatives, their preparation methods, and applications, belonging to the field of pharmaceutical technology. The structure of the benzofuran derivative is as follows: The advantages of this invention are: the IC50 of some compounds (compounds A3, A4, A11, A12, A13, A14, A15, A25, A31, A32, A39) on the inhibitory effects on the proliferation of four tumor cell lines: HeLa, A549, HCT116, and MKN45. 50 All of these compounds showed varying degrees of superiority over the original compound XI-006. Among them, compound A3 exhibited the strongest inhibitory effect on HeLa cells and HCT116 cells, being 10 times and 3.5 times better than XI-006, respectively. Compound A39 showed the strongest inhibitory effect on A549 cells and MKN45 cells, being 2.8 times and 10.8 times better than XI-006, respectively.

Description

Technical Field

[0001] This invention relates to benzofuran derivatives, their preparation methods, and applications, belonging to the field of pharmaceutical technology. Background Technology

[0002] Chemotherapy is a crucial clinical approach to cancer treatment. Chemotherapy drugs inhibit tumor cell proliferation and induce apoptosis by intervening in the biosynthesis and function of tumor nucleic acids and proteins, thereby rapidly controlling tumor development and shrinking the tumor size, especially effective against actively proliferating tumors. Research on the antitumor activity of benzofuran compounds has only recently attracted attention, with 7-(4-methylpiperazin-1-yl)-4-nitro-1-oxobenzofuran (XI-006) being a representative compound. XI-006 can inhibit the expression of the oncogenic protein MDMX, thereby activating the tumor suppressor gene p53, demonstrating good antitumor potential. However, the development of benzofuran derivatives faces numerous challenges, such as poor chemical stability, difficulty in synthesis and structural optimization, potential toxic side effects, and insufficient therapeutic efficacy, with therapeutic efficacy being the primary challenge. Therefore, obtaining drugs with better tumor-suppressive activity based on the benzofuran structure has become an important direction for further research into this class of compounds. Summary of the Invention

[0003] To address the shortcomings of existing technologies, the present invention aims to provide a benzofuran derivative with better tumor-suppressive activity, as well as a method for preparing the derivative and its applications.

[0004] To achieve the above objectives, the present invention adopts the following technical solution:

[0005] The structure of benzofuran derivatives is shown below:

[0006] ;

[0007] Wherein, R1 is selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0008] The preparation method of the aforementioned benzofuran derivative includes the following steps:

[0009] (1) Compound I was prepared using 4-chloro-2-nitroaniline as a raw material;

[0010] (2) Compound II was prepared using compound I as a raw material;

[0011] (3) Compound III was prepared using compound II as a raw material;

[0012] (4) Compound III is dissolved in dichloromethane, and triethylamine and a substituted amine are added, wherein the substituted amine is N-ethylpiperazine, N-isopropylpiperazine, 1-methyl-1,4-diazacycloheptane, 1-ethyl-1,4-diazacycloheptane, thiomorpholine, 2-(tert-butoxycarbonyl)-2,8-diazaspiro[4.5]decane, 1-(2-methoxyethyl)piperazine, 1,4-dioxane- 8-azaspiro[4.5]decane, 8-azaspiro[4.5]decane, 4-(dimethylamino)piperidine, 4-(diethylamino)piperidine, 4-(azacyclobutan-1-yl)piperidine, 4-(pyrrolidine-1-yl)piperidine, 1,4'-bipiperidine, 4-methyl-1,4'-bipiperidine, 3-methylpiperidine, 4,4-difluoropiperidine, 4,4-dimethylpiperidine, 1-pentylpiperazine, 1 -hexylpiperazine, 1-octylpiperazine, 1-(cyclobutylmethyl)piperazine, 1-(cyclopentylmethyl)piperazine, 1-(cyclohexylmethyl)piperazine, 1-(1-methylpiperidin-4-yl)piperazine, 1-(o-tolyl)piperazine, 1-(4-fluorophenyl)piperazine, 1-propionylpiperazine, 1-isobutyrylpiperazine, 1-neopentylpiperazine, 1-cyclopentylpiperazine, 1-cyclohexylpiperazine, 1-( 4-Methoxyphenyl)piperidine, 4-phenylpiperidine, 1-methyl-4-(4-piperidinyl)piperazine, 4-(4-piperidinyl)morpholine, 1-(azacyclobutane-3-yl)piperidine, 1-(azacyclobutane-3-yl)-4-fluoropiperidine or 1,4-diazabicyclo[3.2.2]nonane were reacted with stirring at room temperature. After the reaction was completed, the product was purified and dried to obtain benzofuran derivatives.

[0013] The structures of compounds I, II, and III are shown below:

[0014] , , .

[0015] Preferably, in step (1), the method for preparing compound I is as follows: under ice-water bath conditions, sodium hydroxide is dissolved in anhydrous methanol, and 4-chloro-2-nitroaniline and a 5% sodium hypochlorite aqueous solution are added. The ratio of sodium hydroxide, anhydrous methanol, 4-chloro-2-nitroaniline and sodium hypochlorite aqueous solution is 30.5 mmol: 73 mL: 29.05 mmol: 87.15 mL. The mixture is stirred for 2 h under ice-water bath conditions. After the reaction is completed, the product is purified and dried to obtain compound I.

[0016] Preferably, in step (2), the method for preparing compound II is as follows: compound I is dissolved in concentrated sulfuric acid, and a mixture of fuming nitric acid and concentrated sulfuric acid is slowly added dropwise under ice-water bath conditions. The ratio of the amount of compound I, concentrated sulfuric acid used to dissolve compound I, fuming nitric acid and concentrated sulfuric acid mixed with fuming nitric acid is 22.6 mmol: 20 mL: 29.4 mmol: 3 mL. The mixture is stirred for 30 min under ice-water bath conditions. After the reaction is completed, the product is purified and dried to obtain compound II.

[0017] Preferably, in step (3), the method for preparing compound III is as follows: compound II is dissolved in glacial acetic acid, the ratio of compound II to glacial acetic acid is 9 mmol: 14 mL, the mixture is heated under reflux for 3 h, and after the reaction is completed, the product is purified and dried to obtain compound III.

[0018] Preferably, in step (4), the molar ratio of compound III, triethylamine and substituted amine is 2.3:7:2.8.

[0019] The aforementioned application of benzofuran derivatives in the preparation of anti-cervical cancer drugs, wherein the benzofuran derivatives include:

[0020] , , , , , , , , .

[0021] The aforementioned application of benzofuran derivatives in the preparation of anti-lung cancer drugs, wherein the benzofuran derivatives include:

[0022] , , , , , , , , , , , , , , .

[0023] The aforementioned application of benzofuran derivatives in the preparation of anti-colorectal cancer drugs, wherein the benzofuran derivative is:

[0024] .

[0025] The aforementioned application of benzofuran derivatives in the preparation of anti-gastric cancer drugs, wherein the benzofuran derivatives include:

[0026] , , , , , , , , .

[0027] The advantages of this invention are:

[0028] (1) The benzofuran derivatives A3, A4, A11, A12, A13, A14, A15, A25 and A39 provided by the present invention can significantly inhibit the proliferation of tumor cells HeLa, and the inhibitory effect is significantly better than that of XI-006, showing good anti-tumor activity and having the potential to be developed into anti-cervical cancer drugs.

[0029] (2) The benzofuran derivatives A3, A4, A10, A11, A12, A13, A14, A15, A19, A20, A22, A23, A31, A32 and A39 provided by the present invention can significantly inhibit the proliferation of tumor cells A549, and the inhibitory effect is significantly better than that of XI-006, showing good anti-tumor activity and having the potential to be developed into anti-lung cancer drugs;

[0030] (3) The benzofuran derivative A3 provided by the present invention can significantly inhibit the proliferation of tumor cells HCT116, and the inhibitory effect is significantly better than that of XI-006, showing good anti-tumor activity and has the potential to be developed into an anti-colorectal cancer drug.

[0031] (4) The benzofuran derivatives A3, A4, A11, A12, A13, A14, A15, A25 and A39 provided by the present invention can significantly inhibit the proliferation of tumor cells MKN45, and the inhibitory effect is significantly better than that of XI-006, showing good anti-tumor activity and having the potential to be developed into anti-gastric cancer drugs.

[0032] (5) The preparation method of the above-mentioned benzofuran derivative provided by the present invention is simple, practical and easy to promote. Detailed Implementation

[0033] The present invention will be described in detail below with reference to specific embodiments.

[0034] I. Structure of Benzofuran Derivatives

[0035] The structure of the benzofuran derivative provided by this invention is shown below:

[0036] ;

[0037] Wherein, R1 is selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0038] II. Preparation of benzofuran derivatives

[0039] Example 1

[0040]

[0041] Preparation method of 7-(4-ethylpiperazin-1-yl)-4-nitro-1-oxobenzofuran (compound A1):

[0042] (1) Under ice-water bath conditions, 30.5 mmol of sodium hydroxide was dissolved in 73 mL of anhydrous methanol, 29.05 mmol of 4-chloro-2-nitroaniline was added, followed by 87.15 mL of 5% (w / v) sodium hypochlorite aqueous solution. The mixture was stirred for 2 h under ice-water bath conditions. After the reaction was completed, the product was purified. Specifically, the reaction solution was poured into 75 mL of water, and a solid was precipitated. The solid was filtered, the filter cake was washed with pure water and dried under vacuum at room temperature, and the obtained solid was further purified by normal phase chromatography. The column was filled with 200-300 mesh silica gel, and the mobile phase was a mixture of ethyl acetate and petroleum ether at a volume ratio of 1:50. The eluent was collected, evaporated to dryness, and dried under vacuum at room temperature to obtain compound I.

[0043] (2) 22.6 mmol of compound I was dissolved in 20 mL of concentrated sulfuric acid. Under ice-water bath conditions, a mixture of 29.4 mmol of fuming nitric acid and 3 mL of concentrated sulfuric acid was slowly added dropwise. The mixture was stirred for 30 min under ice-water bath conditions. After the reaction was completed, the product was purified. Specifically, the reaction solution was poured into 20 mL of ice water. A red oily substance precipitated out. The product was extracted three times with ethyl acetate (20 mL × 3). The organic phases were combined, dried, and the solid obtained after removing the solvent was further purified by normal phase chromatography. The column was filled with 200-300 mesh silica gel. The mobile phase was a mixture of ethyl acetate and petroleum ether at a volume ratio of 1:10. The eluent was collected, evaporated to dryness, and then dried under vacuum at room temperature to obtain compound II.

[0044] (3) Dissolve 9 mmol of compound II in 14 mL of glacial acetic acid and heat under reflux for 3 h. After the reaction is complete, purify the product. Specifically, cool the reaction solution to room temperature. A small amount of solid precipitates out. Place it in an ice-water bath and add a small amount of ice water. A large amount of solid precipitates out. Filter the solution. Rinse the filter cake with cold water. Dry the solid under vacuum at room temperature to obtain compound III.

[0045] (4) 2.3 mmol of compound III was dissolved in 10 mL of dichloromethane, 7 mmol of triethylamine and 2.8 mmol of substituted amine N-ethylpiperazine were added, and the mixture was stirred at room temperature for 1 h. After the reaction was completed, the product was purified. Specifically, the solvent was removed, and the obtained solid was purified by normal phase column chromatography with 200-300 mesh silica gel. The mobile phase was a mixture of dichloromethane and methanol at a volume ratio of 30:1. The eluent was collected, evaporated to dryness, and then dried under vacuum at room temperature to obtain a red powder (compound A1) with a yield of 80.2%.

[0046] Compound A1 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0047] 1 H NMR (600MHz, CDCl3) δ 8.34 (d, J=8.6Hz, 1H), 6.26 (d, J =8.6Hz, 1H), 3.48 (t, J =4.9Hz, 4H), 2.72 (q, J =6.2, 5.6Hz, 4H), 2.57-2.52 (m, 2H), 1.16 (t, J =7.2Hz, 3H);

[0048] 13 C NMR (151MHz, CDCl3) δ 147.17, 146.61, 134.88, 131.73, 111.63, 105.50, 52.13, 51.88, 51.88, 51.42, 51.42, 11.88;

[0049] MS (+ESI) m / z: 294.3 (M+H) + ).

[0050] , , .

[0051] Example 2

[0052]

[0053] The preparation method of 7-(4-isopropylpiperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A2) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by N-isopropylpiperazine in equimolar amounts. A red powder (denoted as compound A2) was obtained with a yield of 74.5%.

[0054] Compound A2 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0055] 1 H NMR (600MHz, CDCl3) δ 8.33 (d, J =8.6Hz, 1H), 6.25 (d, J =8.6Hz, 1H), 3.52-3.42 (m, 4H), 2.82 (p, J =6.1, 5.7Hz, 1H), 2.77 (q, J =4.7Hz, 4H), 1.11 (dd, J =6.6, 3.2 Hz, 6H).

[0056] 13C NMR (151MHz, CDCl3) δ 147.20, 146.66, 134.93, 131.82, 111.61, 105.38, 54.64, 51.80, 49.80, 48.33, 47.91, 18.41, 18.39;

[0057] MS (+ESI) m / z: 308.2 (M+H) + ).

[0058] Example 3

[0059]

[0060] The preparation method of 7-(4-methyl-1,4-diazacycloheptane-1-yl)-4-nitro-1-oxobenzofuran (compound A3) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1-methyl-1,4-diazacycloheptane. A red powder (denoted as compound A3) was obtained with a yield of 45.9%.

[0061] Compound A3 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0062] 1 H NMR (600MHz, CDCl3) δ 8.04 (d, J=9.0Hz, 1H), 6.04 (d, J=8.9Hz, 1H), 4.27-4.17 (m, 1H), 3.97 (m, 1H), 3.86-3.81 (m, 1H), 3.80-3.72 (m, 1H), 2.88 (q, J=5.7, 5.2Hz, 2H), 2.70 (dt, J=16.8, 5.5Hz, 2H), 2.45 (s, 3H), 2.14 (dp, J=26.3, 5.6Hz, 2H);

[0063] 13 C NMR (151MHz, CDCl3) δ 147.67, 145.16, 135.23, 132.51, 102.71, 101.65, 58.25, 56.93, 51.98, 46.80, 31.51, 30.14;

[0064] MS (+ESI) m / z: 294.1 (M+H) + ).

[0065] Example 4

[0066]

[0067] The preparation method of 7-(4-ethyl-1,4-diazacycloheptane-1-yl)-4-nitro-1-oxobenzofuran (compound A4) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1-ethyl-1,4-diazacycloheptane. A red powder (denoted as compound A4) was obtained with a yield of 40.2%.

[0068] Compound A4 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0069] 1 H NMR (600MHz, CDCl3) δ 8.03 (d, J=9.0Hz, 1H), 6.05 (d, J=9.0Hz, 1H), 4.19 (s, 2H), 3.99 (s, 2H), 2.93 (dt, J=10.5, 4.8Hz, 2H), 2 .76 (dt, J=20.8, 5.6Hz, 2H), 2.64 (p, J=6.9Hz, 2H), 2.13 (dd, J=11.4, 5.7Hz, 2H), 1.12 (q, J=7.3Hz, 3H);

[0070] 13 C NMR (151MHz, CDCl3) δ 147.66, 145.17, 135.25, 132.55, 102.75, 101.64, 55.38, 54.10, 53.37, 52.21, 52.15, 51.83, 12.29;

[0071] MS (+ESI) m / z: 308.4 (M+H) + ).

[0072] Example 5

[0073]

[0074] The preparation method of 7-(thiomorpholino)-4-nitro-1-oxobenzofuran (compound A5) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by an equimolar amount of thiomorpholino. A brown powder (denoted as compound A5) is obtained with a yield of 76.6%.

[0075] Compound A5 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0076] 1H NMR (600MHz, CDCl3) δ 8.34 (d, J=8.5Hz, 1H), 6.30 (d, J=8.5Hz, 1H), 3.66 (dd, J=6.4, 3.6Hz, 4H), 2.95-2.89 (m, 4H);

[0077] 13 C NMR (151MHz, CDCl3) δ 147.10, 147.04, 134.62, 111.68, 106.32, 53.93, 53.93, 27.25, 27.25;

[0078] MS (+ESI) m / z: 283.4 (M+H) + ).

[0079] Example 6

[0080]

[0081] The preparation method of 7-(2-(tert-butoxycarbonyl)-2,8-diazaspiro[4.5]decane-8-yl)-4-nitro-1-oxobenzofuran (compound A6) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 2-(tert-butoxycarbonyl)-2,8-diazaspiro[4.5]decane. A red powder (denoted as compound A6) was obtained with a yield of 62.7%.

[0082] Compound A6 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0083] 1 H NMR (600MHz, CDCl3) δ 8.32 (d, J=8.5Hz, 1H), 6.26 (d, J=9.6Hz, 1H), 3.54-3.43 (m, 4H), 3.38-3.26 (m, 4H), 1.84 (q, J=5.1, 3.3Hz, 6H), 1.49 (s, 9H);

[0084] 13 C NMR (151MHz, CDCl3) δ 147.19, 146.86, 134.91, 134.91, 127.07, 111.61, 105.47, 79.55, 54.46, 49 .49, 49.49, 44.09, 43.88, 39.36, 36.78, 34.10, 34.10, 28.54, 28.54, 28.54;

[0085] MS (+ESI) m / z: 420.3 (M+H) + ).

[0086] Example 7

[0087]

[0088] The preparation method of 7-(4-(2-methoxyethyl)piperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A7) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by 1-(2-methoxyethyl)piperazine in equimolar amounts. A red powder (denoted as compound A7) was obtained with a yield of 40.7%.

[0089] Compound A7 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0090] 1 H NMR (600MHz, CDCl3) δ 8.34 (d, J=8.6Hz, 1H), 6.25 (d, J=8.6Hz, 1H), 3.58 (t, J=5.2Hz, 2H), 3.49 (t, J=5.0Hz, 4H), 3.40 (s, 3H), 2.78 (s, 4H), 2.71 (q, J=4.9Hz, 2H);

[0091] 13 C NMR (151MHz, CDCl3) δ 147.16, 146.62, 134.88, 131.73, 111.63, 105.48, 69.83, 59.02, 57.66, 53.02, 52.61, 51.26, 49.27;

[0092] MS (+ESI) m / z: 324.2 (M+H) + ).

[0093] Example 8

[0094]

[0095] The preparation method of 7-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)-4-nitro-1-oxobenzofuran (compound A8) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1,4-dioxa-8-azaspiro[4.5]decane. A red powder (denoted as compound A8) was obtained with a yield of 70.1%.

[0096] Compound A8 1H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0097] 1 H NMR (600MHz, CDCl3) δ 8.33 (d, J=8.6Hz, 1H), 6.26 (d, J=8.6Hz, 1H), 4.05 (d, J=5.8Hz, 4H), 3.56-3.48 (m, 4H), 2.01-1.96 (m, 4H);

[0098] 13 C NMR (151MHz, CDCl3) δ 147.18, 146.74, 134.90, 127.11, 111.61, 105.83, 105.66, 64.64, 64.64, 49.98, 49.98, 34.65, 34.65;

[0099] MS (+ESI) m / z: 323.4 (M+H) + ).

[0100] Example 9

[0101]

[0102] The preparation method of 7-(8-azaspiro[4.5]decane-8-yl)-4-nitro-1-oxobenzofuran (compound A9) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 8-azaspiro[4.5]decane. A brown powder (denoted as compound A9) was obtained with a yield of 72.5%.

[0103] Compound A9 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0104] 1 H NMR (600MHz, DMSO) δ 8.38 (d, J=8.9Hz, 1H), 6.48 (d, J=8.9Hz, 1H), 3.56-3.39 (m, 4H), 1.64-1.59 (m, 8H), 1.49 (dtd, J=7.2, 5.1, 4.1, 1.9Hz, 4H);

[0105] 13C NMR (151MHz, DMSO) δ 148.14, 147.59, 136.60, 124.37, 112.24, 106.20, 50.01, 40.78, 40.78, 37.63, 37.63, 37.06, 37.06, 24.28, 24.28;

[0106] MS (+ESI) m / z: 319.3 (M+H) + ).

[0107] Example 10

[0108]

[0109] The preparation method of 7-(4-(dimethylamino)piperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A10) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 4-(dimethylamino)piperidine. A red powder (denoted as compound A10) was obtained with a yield of 69.2%.

[0110] Compound A10 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0111] 1 H NMR (600MHz, CDCl3) δ 8.30 (dd, J=8.6, 1.6Hz, 1H), 6.24 (dd, J=8.7, 1.5Hz, 1H), 3.78 (d, J=12.8Hz, 2H), 3.08 (t , J=12.3Hz, 2H), 2.36-2.34 (m, 6H), 2.02 (d, J=13.1Hz, 2H), 1.78 (qd, J=12.4, 4.0Hz, 2H);

[0112] 13 C NMR (151MHz, DMSO) δ 148.07, 147.40, 136.60, 124.80, 112.31, 106.40, 60.54, 50.98, 50.98, 41.85, 41.85, 28.25, 28.25;

[0113] MS (+ESI) m / z: 308.3 (M+H) + ).

[0114] Example 11

[0115]

[0116] The preparation method of 7-(4-(diethylamino)piperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A11) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by an equimolar amount of 4-(diethylamino)piperidine. A red powder (denoted as compound A11) is obtained with a yield of 80.4%.

[0117] Compound A11 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0118] 1 H NMR (600MHz, MeOD) δ 8.38 (d, J=8.7Hz, 1H), 6.47-6.44 (m, 1H), 3.89 (dp, J=13.1, 2.4Hz, 2H), 3.13 (td, J=12.8, 1.9Hz, 2H), 2.96- 2.90 (m, 1H), 2.72 (q, J=7.1Hz, 4H), 2.05-2.00 (m, 2H), 1.81 (tt, J=12.5, 6.3Hz, 2H), 1.13 (t, J=7.1Hz, 6H);

[0119] 13 C NMR (151MHz, CDCl3) δ 147.23, 146.77, 134.97, 126.76, 111.53, 105.34, 56.92, 51.64, 51.64, 43.62, 43.62, 28.12, 28.12, 13.49, 13.49;

[0120] MS (+ESI) m / z: 336.2 (M+H) + ).

[0121] Example 12

[0122]

[0123] The preparation method of 7-(4-(azacyclobutan-1-yl)piperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A12) was basically the same as in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 4-(azacyclobutan-1-yl)piperidine. A red powder (denoted as compound A12) was obtained with a yield of 61.0%.

[0124] Compound A12 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0125] 1 H NMR (600MHz, DMSO) δ 8.38 (d, J =8.8Hz, 1H), 6.48 (d, J =8.9Hz, 1H), 3.68 (dt, J =13.3, 4.7Hz, 2H), 3.22 (ddd, J =12.9, 9.6, 3.1Hz, 4H), 2.47 (s, 1H), 1.99 (p, J =7.3Hz, 2H), 1.83-1.78 (m, 2H), 1.40 (ddd, J =13.1, 8.8, 3.7Hz, 2H), 1.23 (s, 2H);

[0126] 13 C NMR (151MHz, DMSO) δ 148.06, 147.49, 136.59, 124.82, 112.34, 106.43, 61.71, 52.81, 52.81, 49.59, 49.59, 29.47, 28.11, 16.78, 16.78;

[0127] MS (+ESI) m / z: 320.3 (M+H) + ).

[0128] Example 13

[0129]

[0130] The preparation method of 7-(4-(pyrrolidine-1-yl)piperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A13) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 4-(pyrrolidine-1-yl)piperidine. A red powder (denoted as compound A13) was obtained with a yield of 77.5%.

[0131] Compound A13 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0132] 1H NMR (600MHz, CDCl3) δ 8.32 (d, J=8.6Hz, 1H), 6.24 (d, J=8.7Hz, 1H), 3.77 (dt, J=13.8, 4.1Hz, 2H), 3.20 (ddd, J =13.2, 10.7, 2.8Hz, 2H), 2.71 (s, 4H), 2.13 (dt, J=13.7, 3.7Hz, 2H), 1.93-1.85 (m, 6H);

[0133] 13 C NMR (151MHz, CDCl3) δ 147.26, 146.76, 134.99, 126.60, 111.51, 105.20, 60.09, 51.44, 51.44, 50.28, 50.28, 30.62, 30.62, 23.34, 23.34;

[0134] MS (+ESI) m / z: 334.4 (M+H) + ).

[0135] Example 14

[0136]

[0137] The preparation method of 7-(1,4'-bipiperidine-1'-yl)-4-nitro-1-oxobenzofuran (compound A14) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced with 1,4'-bipiperidine in equal molar amounts. A red powder (denoted as compound A14) was obtained with a yield of 59.8%.

[0138] Compound A14 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0139] 1 H NMR (600MHz, CDCl3) δ 8.32 (d, J=8.6Hz, 1H), 6.25 (d, J=8.6Hz, 1H), 3.82 (dq, J=13.1, 2.4Hz, 2H), 3.11 (td, J=12.5, 2.4Hz, 2H), 2.66 (s, 4H), 2.09 (d, J=12.7Hz, 2H), 1.94-1.86 (m, 2H), 1.71 (q, J=5.7Hz, 4H), 1.52 (p, J=6.7, 6.2Hz, 2H);

[0140] 13C NMR (151MHz, CDCl3) δ 147.20, 146.66, 134.95, 126.83, 111.53, 105.43, 61.55, 51.45, 51.45, 50.29, 50.29, 27.44, 27.44, 25.91, 25.91, 24.38;

[0141] MS (+ESI) m / z: 348.3 (M+H) + ).

[0142] Example 15

[0143]

[0144] The preparation method of 7-(4-methyl-1,4'-bipiperidine-1'-yl)-4-nitro-1-oxobenzofuran (compound A15) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by equimolar amounts of 4-methyl-1,4'-bipiperidine. A red powder (denoted as compound A15) was obtained with a yield of 72.1%.

[0145] Compound A15 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0146] 1 H NMR (600MHz, CDCl3) δ 8.32 (d, J=8.6Hz, 1H), 6.25 (d, J=8.7Hz, 1H), 3.85-3.80 (m, 2H), 3.11 (td, J=12.4, 2.4Hz, 2H), 3.05 (d, J=11.1Hz, 2H), 2.73 (s, 1H), 2.33 (d, J =11.8Hz, 2H), 2.10-2.06 (m, 2H), 1.93-1.86 (m, 2H), 1.77-1.72 (m, 2H), 1.44 (q, J=10.7, 9.9Hz, 1H), 1.37-1.31 (m, 2H), 0.97 (d, J=6.6Hz, 3H);

[0147] 13 C NMR (151MHz, CDCl3) δ 147.20, 146.66, 134.94, 126.84, 111.53, 105.41, 61.25, 51.43, 51.43, 49.76, 49.76, 34.22, 34.22, 30.85, 27.64, 27.64, 21.75;

[0148] MS (+ESI) m / z: 362.4 (M+H) + ).

[0149] Example 16

[0150]

[0151] The preparation method of 7-(3-methylpiperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A16) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by an equimolar amount of 3-methylpiperidine. A red powder (denoted as compound A16) was obtained with a yield of 66.3%.

[0152] Compound A16 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0153] 1 H NMR (600MHz, CDCl3) δ 8.33 (d, J=8.6Hz, 1H), 6.22 (d, J=8.7Hz, 1H), 3.72 (dd, J=12.8, 2.8Hz, 1H), 3.65 (ddt, J=12.6, 3.9, 2.0Hz, 1H), 3.08-3.02 (m, 1H), 2.73 (dd, J=12.5, 10.5Hz, 1H), 1.97-1.82 (m, 4H), 1.26-1.20 (m, 1H), 1.01 (d, J=6.4Hz, 3H);

[0154] 13 C NMR (151MHz, CDCl3) δ 147.38, 147.05, 135.13, 126.09, 111.45, 104.87, 59.32, 52.38, 32.11, 31.16, 25.03, 19.00;

[0155] MS (+ESI) m / z: 279.1 (M+H) + ).

[0156] Example 17

[0157]

[0158] The preparation method of 7-(4,4-difluoropiperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A17) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by equimolar amounts of 4,4-difluoropiperidin. A red powder (denoted as compound A17) was obtained with a yield of 39.7%.

[0159] Compound A17 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0160] 1 H NMR (600MHz, CDCl3) δ 8.35 (d, J=8.5Hz, 1H), 6.33 (d, J=8.4Hz, 1H), 3.51 (t, J=5.7Hz, 4H), 2.30 (tt, J=13.0, 5.8Hz, 4H);

[0161] 13 C NMR (151MHz, CDCl3) δ 135.38, 131.36, 124.58, 124.32, 124.29, 123.78, 123.22, 31.99, 31.99, 23.38, 23.38;

[0162] MS (+ESI) m / z: 301.2 (M+H) + ).

[0163] Example 18

[0164]

[0165] The preparation method of 7-(4,4-dimethylpiperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A18) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by equimolar amounts of 4,4-dimethylpiperidin. A red powder (denoted as compound A18) was obtained with a yield of 71.5%.

[0166] Compound A18 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0167] 1 H NMR (600MHz, CDCl3) δ 8.33 (d, J=8.6Hz, 1H), 6.24 (d, J=8.7Hz, 1H), 3.49-3.38 (m, 4H), 1.66-1.63 (m, 4H), 1.08 (s, 6H);

[0168] 13 C NMR (151MHz, CDCl3) δ 147.37, 147.10, 135.12, 126.13, 111.47, 104.89, 48.66, 48.66, 38.02, 38.02, 28.51, 27.76, 27.76;

[0169] MS (+ESI) m / z: 293.2 (M+H) + ).

[0170] Example 19

[0171]

[0172] The preparation method of 7-(4-pentylpiperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A19) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-pentylpiperazine in equal molar amounts. A red powder (denoted as compound A19) was obtained with a yield of 63.4%.

[0173] Compound A19 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0174] 1 H NMR (600MHz, CDCl3) δ 8.33 (d, J=8.6Hz, 1H), 6.25 (d, J=8.6Hz, 1H), 3.47 (t, J=4.9Hz, 4H), 2.70 (q, J=5.2Hz, 4H), 2 .45 (dd, J=9.2, 6.3Hz, 2H), 1.55 (t, J=7.6Hz, 2H), 1.38-1.32 (m, 4H), 0.93 (t, J=7.0Hz, 3H);

[0175] 13 C NMR (151MHz, CDCl3) δ 147.17, 146.61, 134.87, 127.23, 111.63, 105.46, 58.37, 58.37, 52.30, 52.30, 51.41, 29.60, 26.35, 22.58, 14.04;

[0176] MS (+ESI) m / z: 336.2 (M+H) + ).

[0177] Example 20

[0178]

[0179] The preparation method of 7-(4-hexylpiperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A20) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-hexylpiperazine in equal molar amounts. A red powder (denoted as compound A20) was obtained with a yield of 69.9%.

[0180] Compound A20 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0181] 1 H NMR (600MHz, CDCl3) δ 8.34 (d, J=8.6Hz, 1H), 6.26 (d, J=8.6Hz, 1H), 3.48 (t, J=4.9Hz, 4H), 2.71 (d, J=14.2Hz, 4H) , 2.46 (q, J=7.3Hz, 2H), 1.55 (t, J=7.6Hz, 2H), 1.34 (q, J=5.4Hz, 6H), 0.92 (t, J=6.7Hz, 3H);

[0182] 13 C NMR (151MHz, CDCl3) δ 147.17, 146.60, 134.88, 127.23, 111.63, 105.47, 58.41, 58.41, 52.31, 52.31, 51.39, 31.72, 27.10, 26.62, 22.60, 14.05;

[0183] MS (+ESI) m / z: 350.3 (M+H) + ).

[0184] Example 21

[0185]

[0186] The preparation method of 7-(4-octylpiperazin-1-yl)-4-nitro-1-oxobenzofuran (compound A21) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-octylpiperazine in equal molar amounts. A red powder (denoted as compound A21) is obtained with a yield of 60.1%.

[0187] Compound A21 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0188] 1H NMR (600MHz, CDCl3) δ 8.34 (d, J=8.6Hz, 1H), 6.25 (d, J=8.6Hz, 1H), 3.48 (t, J=4.9Hz, 4H), 2.70 (dt, J=11.6, 4.4Hz, 4H), 2.45 (d t, J=12.1, 7.5Hz, 2H), 1.55 (t, J=7.5Hz, 2H), 1.31 (ddd, J=23.9, 12.8, 6.7Hz, 10H), 0.91 (t, J=6.9Hz, 3H);

[0189] 13 C NMR (151MHz, CDCl3) δ 147.17, 146.60, 134.87, 127.26, 111.64, 105.47, 58.40, 58.40, 52.29, 52.29, 51.38, 31.83, 29.48, 29.24, 27.43, 26.64, 22.66, 14.11;

[0190] MS (+ESI) m / z: 378.5 (M+H) + ).

[0191] Example 22

[0192]

[0193] The preparation method of 7-(4-(cyclobutylmethyl)piperazin-1-yl)-4-nitro-1-oxobenzofuran (compound A22) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by 1-(cyclobutylmethyl)piperazine in equimolar amounts. A red powder (denoted as compound A22) was obtained with a yield of 47.0%.

[0194] Compound A22 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0195] 1 H NMR (600MHz, CDCl3) δ 8.33 (d, J=8.6Hz, 1H), 6.24 (d, J=8.6Hz, 1H), 3.45 (t, J=4.9Hz, 4H), 2.67 (q, J=4.9Hz, 4H), 2.60-2.56 (m, 1H) , 2.52 (d, J=7.0Hz, 2H), 2.13-2.09 (m, 2H), 1.98-1.93 (m, 1H), 1.89-1.84 (m, 1H), 1.74 (td, J=8.6, 2.5Hz, 2H);

[0196] 13 C NMR (151MHz, CDCl3) δ 147.20, 146.69, 134.90, 127.10, 111.60, 105.33, 64.75, 52.39, 52.39, 51.49, 51.49, 33.61, 27.73, 27.73, 18.85;

[0197] MS (+ESI) m / z: 334.3 (M+H) + ).

[0198] Example 23

[0199]

[0200] The preparation method of 7-(4-(cyclopentylmethyl)piperazin-1-yl)-4-nitro-1-oxobenzofuran (compound A23) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-(cyclopentylmethyl)piperazine in equimolar amounts. A red powder (denoted as compound A23) was obtained with a yield of 48.5%.

[0201] Compound A23 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0202] 1 H NMR (600MHz, CDCl3) δ 8.34 (d, J=8.6Hz, 1H), 6.24 (d, J=8.6Hz, 1H), 3.47 (t, J=4.8Hz, 4H), 2.69 (d, J=9.6Hz, 4H), 2.37 (dd, J=11.6 , 7.4Hz, 2H), 2.15-2.10 (m, 1H), 1.82-1.77 (m, 2H), 1.70-1.57 (m, 4H), 1.25 (ddd, J=14.9, 11.9, 7.3Hz, 2H);

[0203] 13 C NMR (151MHz, CDCl3) δ 147.20, 146.66, 134.90, 111.62, 105.36, 64.10, 64.10, 52.54, 52.54, 51.44, 36.95, 31.30, 31.30, 25.18, 25.18;

[0204] MS (+ESI) m / z: 348.4 (M+H) + ).

[0205] Example 24

[0206]

[0207] The preparation method of 7-(4-(cyclohexylmethyl)piperazin-1-yl)-4-nitro-1-oxobenzofuran (compound A24) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1-(cyclohexylmethyl)piperazine. A red powder (denoted as compound A24) was obtained with a yield of 53.6%.

[0208] Compound A24 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0209] 1 H NMR (600MHz, CDCl3) δ 8.33 (d, J=8.6Hz, 1H), 6.24 (d, J=8.6Hz, 1H), 3.46 (t, J=4.9Hz, 4H), 2.65 (dd, J=8.3, 4.4Hz, 4H), 2.24 (d, J=7.6 Hz, 2H), 1.80-1.72 (m, 4H), 1.54 (ddt, J=11.1, 7.4, 3.7Hz, 1H), 1.29-1.21 (m, 4H), 0.92 (dd, J=12.0, 3.6Hz, 2H);

[0210] 13 C NMR (151MHz, CDCl3) δ 147.22, 146.74, 134.92, 111.60, 105.24, 65.23, 65.23, 52.74, 52.74, 51.60, 34.99, 31.77, 31.77, 26.75, 26.08, 26.08;

[0211] MS (+ESI) m / z: 362.3 (M+H) + ).

[0212] Example 25

[0213]

[0214] The preparation method of 7-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-4-nitro-1-oxobenzofuran (compound A25) was basically the same as in Example 1, except that in step (4), N-ethylpiperazine was replaced with 1-(1-methylpiperidin-4-yl)piperazine in equimolar amounts. A red powder (denoted as compound A25) was obtained with a yield of 53.6%.

[0215] Compound A251 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0216] 1 H NMR (600MHz, CDCl3) δ 8.34 (d, J=8.6Hz, 1H), 6.24 (d, J=8.7Hz, 1H), 3.52-3.38 (m, 4H), 3.04 (d, J=11.2Hz, 2H), 2.81 (dd, J=5.8 , 3.6Hz, 4H), 2.46-2.40 (m, 1H), 2.38 (d, J=5.9Hz, 3H), 2.13 (s, 2H), 1.87 (s, 2H), 1.76 (t, J=12.0Hz, 2H);

[0217] 13 C NMR (151MHz, CDCl3) δ 147.19, 146.61, 134.91, 127.14, 111.61, 105.39, 54.99, 51.88, 51.88, 48.41, 48.41, 48.41, 48.41, 45.72, 27.75, 27.75;

[0218] MS (+ESI) m / z: 363.5 (M+H) + ).

[0219] Example 26

[0220]

[0221] The preparation method of 7-(4-(o-tolyl)piperazin-1-yl)-4-nitro-1-oxobenzofuran (compound A26) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by 1-(o-tolyl)piperazine in equimolar amounts. A red powder (denoted as compound A26) was obtained with a yield of 76.9%.

[0222] Compound A26 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0223] 1H NMR (600MHz, CDCl3) δ 8.37 (d, J=8.5Hz, 1H), 7.24 (dd, J=9.3, 7.3Hz, 2H), 7.12-7.07 (m, 2H), 6.33 ( d, J=8.6Hz, 1H), 3.67-3.54 (m, 4H), 3.19 (dd, J=5.9, 3.7Hz, 4H), 2.38 (s, 3H);

[0224] 13 C NMR (151MHz, CDCl3) δ 150.25, 147.17, 146.79, 134.82, 132.70, 131.59, 131.33, 126.87, 124.22, 119.41, 111.73, 105.69, 52.04, 52.04, 51.19, 51.19, 17.85;

[0225] MS (+ESI) m / z: 356.2 (M+H) + ).

[0226] Example 27

[0227] The preparation method of 7-(4-(4-fluorophenyl)piperazin-1-yl)-4-nitro-1-oxobenzofuran (compound A27) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1-(4-fluorophenyl)piperazine. A red powder (denoted as compound A27) was obtained with a yield of 50.7%.

[0228] Compound A27 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0229] 1 H NMR (600MHz, DMSO) δ 8.44 (d, J=8.7Hz, 1H), 7.13-6.99 (m, 5H), 6.58 (dd, J=8.9, 5.0Hz, 1H), 3.62 (t, J=4.9Hz, 4H), 3.29 (t, J=5.0Hz, 4H);

[0230] 13 C NMR (151MHz, DMSO) δ 157.59, 147.98, 147.77, 147.10, 136.52, 125.72, 118.08, 118.03, 115.95, 115.80, 112.54, 106.93, 51.31, 51.31, 49.10, 49.10;

[0231] MS (+ESI) m / z: 360.2 (M+H) + ).

[0232] Example 28

[0233]

[0234] The preparation method of 7-(4-propionylpiperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A28) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-propionylpiperazine in equal molar amounts. A red powder (denoted as compound A28) was obtained with a yield of 29.7%.

[0235] Compound A28 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0236] 1 H NMR (600MHz, DMSO) δ 8.43 (d, J=8.7Hz, 1H), 6.53 (d, J=8.8Hz, 1H), 3.65 (t, J=5.0Hz, 4H), 3.48 (dd, J=30.5, 6.4Hz, 4H), 2.38 (q, J=7.3Hz, 2H), 1.01 (t, J=7.3Hz, 3H);

[0237] 13 C NMR (151MHz, DMSO) δ 172.17, 147.95, 147.04, 136.51, 125.74, 112.50, 106.84, 51.65, 50.98, 44.55, 40.84, 25.92, 9.72;

[0238] MS (+ESI) m / z: 322.2 (M+H) + ).

[0239] Example 29

[0240]

[0241] The preparation method of 7-(4-isobutyrylpiperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A29) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-isobutyrylpiperazine in equal molar amounts. A red powder (denoted as compound A29) was obtained with a yield of 32.2%.

[0242] Compound A29 1 H NMR, 13The specific detection results of C NMR and MS (+ESI) are as follows:

[0243] 1 H NMR (600MHz, DMSO) δ 8.44 (d, J=8.7Hz, 1H), 6.53 (d, J=8.8Hz, 1H), 3.68 (dt, J=37.2, 5.2Hz, 4H), 3.48 (dt, J=31.6, 4.7Hz, 4H), 2.93 (p, J=6.9Hz, 1H), 1.02 (d, J=6.7Hz, 6H);

[0244] 13 C NMR (151MHz, DMSO) δ 175.16, 147.94, 147.01, 136.51, 125.84, 112.52, 106.93, 51.86, 51.04, 44.65, 40.96, 29.44, 19.82, 19.82;

[0245] MS (+ESI) m / z: 336.4 (M+H) + ).

[0246] Example 30

[0247]

[0248] The preparation method of 7-(4-neopentylpiperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A30) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-neopentylpiperazine in equal molar amounts. A red powder (denoted as compound A30) was obtained with a yield of 29.7%.

[0249] Compound A30 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0250] 1 H NMR (600MHz, DMSO) δ 8.44 (d, J=8.7Hz, 1H), 6.53 (d, J=8.8Hz, 1H), 3.80-3.70 (m, 4H), 3.54-3.43 (m, 4H), 1.23 (d, J=3.6Hz, 9H);

[0251] 13C NMR (151MHz, DMSO) δ 175.86, 147.96, 147.01, 136.53, 125.88, 112.51, 106.91, 51.47, 51.47, 40.53, 38.57, 38.57, 28.48, 28.48, 28.48;

[0252] MS (+ESI) m / z: 350.3 (M+H) + ).

[0253] Example 31

[0254]

[0255] The preparation method of 7-(4-cyclopentylpiperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A31) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-cyclopentylpiperazine in equal molar amounts. A red powder (denoted as compound A31) was obtained with a yield of 64.5%.

[0256] Compound A31 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0257] 1 H NMR (600MHz, CDCl3) δ 8.34 (d, J=8.6Hz, 1H), 6.25 (d, J=8.6Hz, 1H), 3.48 (t, J=4.9Hz, 4H), 2.80-2.75 (m, 4H), 2.65 (p, J=11.1, 9. 7Hz, 1H), 1.96-1.91 (m, 2H), 1.75 (ddd, J=9.9, 7.3, 5.0Hz, 2H), 1.63-1.60 (m, 2H), 1.46 (d, J=10.2Hz, 2H);

[0258] 13 C NMR (151MHz, CDCl3) δ 147.17, 146.56, 134.88, 131.73, 111.64, 105.41, 67.20, 51.37, 51.37, 51.37, 51.37, 30.33, 30.33, 24.09, 24.09;

[0259] MS (+ESI) m / z: 334.2 (M+H) + ).

[0260] Example 32

[0261]

[0262] The preparation method of 7-(4-cyclohexylpiperazine-1-yl)-4-nitro-1-oxobenzofuran (compound A32) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by 1-cyclohexylpiperazine in equal molar amounts. A red powder (denoted as compound A32) was obtained with a yield of 71.3%.

[0263] Compound A32 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0264] 1 H NMR (600MHz, CDCl3) δ 8.33 (d, J=8.6Hz, 1H), 6.24 (d, J=8.7Hz, 1H), 3.52-3.41 (m, 4H), 2.86-2.81 (m, 4H), 2.42-2.36 (m , 1H), 1.92-1.84 (m, 4H), 1.67 (t, J=6.7Hz, 1H), 1.29-1.25 (m, 4H), 1.15 (td, J=12.2, 3.7Hz, 1H);

[0265] 13 C NMR (151MHz, CDCl3) δ 147.22, 146.70, 134.93, 131.83, 111.59, 105.26, 63.51, 52.05, 52.05, 48.28, 48.28, 28.94, 28.94, 26.21, 25.77, 25.77;

[0266] MS (+ESI) m / z: 348.3 (M+H) + ).

[0267] Example 33

[0268]

[0269] The preparation method of 7-(4-(4-methoxyphenyl)piperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A33) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1-(4-methoxyphenyl)piperidine. A red powder (denoted as compound A33) was obtained with a yield of 56.7%.

[0270] Compound A33 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0271] 1 H NMR (600MHz, DMSO) δ 8.41 (d, J=8.8Hz, 1H), 7.19 (dd, J=9.0, 2.6Hz, 2H), 6.90-6.87 (m, 2H), 6.54 (d, J=8.9Hz, 1H), 3.91 (dq, J=11.6, 2.4Hz , 2H), 3.73 (s, 3H), 3.31-3.25 (m, 2H), 2.84 (tt, J=12.1, 3.8Hz, 1H), 1.94-1.90 (m, 2H), 1.79 (qd, J=12.7, 3.7Hz, 2H);

[0272] 13 C NMR (151MHz, DMSO) δ 158.22, 148.13, 147.60, 137.58, 136.64, 128.06, 128.06, 124.72, 114. 33, 114.33, 112.35, 106.46, 55.48, 55.48, 52.45, 40.51, 33.26, 33.26;

[0273] MS (+ESI) m / z: 371.1 (M+H) + ).

[0274] Example 34

[0275]

[0276] The preparation method of 7-(4-phenylpiperidin-1-yl)-4-nitro-1-oxobenzofurazan (compound A34) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by equimolar amounts of 4-phenylpiperidin. A red powder (denoted as compound A34) was obtained with a yield of 80.6%.

[0277] Compound A34 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0278] 1 H NMR (600MHz, CDCl3) δ 8.36 (d, J=8.6Hz, 1H), 7.39-7.35 (m, 2H), 7.29-7.24 (m, 3H), 6.30 (d, J=8.6Hz, 1H), 3.94-3 .84 (m, 2H), 3.24 (td, J=12.2, 3.4Hz, 2H), 2.85 (tt, J=11.5, 4.5Hz, 1H), 2.13-2.01 (m, 4H);

[0279] 13 C NMR (151MHz, CDCl3) δ 147.28, 147.07, 144.39, 135.01, 135.01, 128.74, 128.74, 126.84, 126.75, 126.75, 111.60, 105.34, 52.58, 52.58, 41.71, 32.74, 32.74;

[0280] MS (+ESI) m / z: 341.2 (M+H) + ).

[0281] Example 35

[0282]

[0283] The preparation method of 7-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A35) was basically the same as in Example 1, except that in step (4), N-ethylpiperazine was replaced with 1-methyl-4-(4-piperidinyl)piperazine in equimolar amounts. A red powder (denoted as compound A35) was obtained with a yield of 45.8%.

[0284] Compound A35 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0285] 1 H NMR (600MHz, DMSO) δ 8.39 (d, J=8.8Hz, 1H), 6.50 (d, J=8.9Hz, 1H), 3.82 (d, J=12.9Hz, 2H), 3.19-3.13 (m, 2H), 3.05 (d , J=7.3Hz, 1H), 2.64 (s, 8H), 2.39 (s, 3H), 1.93 (d, J=13.4Hz, 2H), 1.60 (tt, J=12.1, 6.3Hz, 2H);

[0286] 13 C NMR (151MHz, DMSO) δ 148.05, 147.32, 136.60, 124.96, 112.35, 106.53, 53.92, 53.92, 50.94, 50.94, 47.12, 40.51, 40.51, 29.45, 28.05, 28.05;

[0287] MS (+ESI) m / z: 363.5 (M+H) + ).

[0288] Example 36

[0289]

[0290] The preparation method of 7-(4-morpholinylpiperidin-1-yl)-4-nitro-1-oxobenzofuran (compound A36) is basically the same as that in Example 1, except that in step (4), N-ethylpiperazine is replaced by equimolar amounts of 4-(4-piperidinyl)morpholine. A red powder (denoted as compound A36) was obtained with a yield of 50.6%.

[0291] Compound A36 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0292] 1 H NMR (600MHz, DMSO) δ 8.39 (d, J=8.9Hz, 1H), 6.49 (d, J=8.9Hz, 1H), 3.84-3.78 (m, 2H), 3.58 (q, J=7.6, 6.0Hz, 4H) , 3.20-3.13 (m, 2H), 2.48 (d, J=4.9Hz, 4H), 1.97-1.91 (m, 2H), 1.58 (td, J=11.5, 7.8Hz, 2H);

[0293] 13 C NMR (151MHz, DMSO) δ 148.07, 147.37, 136.60, 124.81, 112.32, 106.40, 66.99, 66.99, 60.17, 51.00, 51.00, 49.90, 49.90, 28.15, 28.15;

[0294] MS (+ESI) m / z: 350.2 (M+H) + ).

[0295] Example 37

[0296]

[0297] The preparation method of 7-(3-(piperidin-1-yl)azacyclobutane-1-yl)-4-nitro-1-oxobenzofuran (compound A37) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1-(azacyclobutane-3-yl)piperidine. A red powder (denoted as compound A37) was obtained with a yield of 46.0%.

[0298] Compound A37 1 H NMR, 13The specific detection results of C NMR and MS (+ESI) are as follows:

[0299] 1 H NMR (600MHz, DMSO) δ 8.33 (d, J=9.1Hz, 1H), 5.90 (d, J=9.1Hz, 1H), 4.77 (d, J=24.3Hz, 1H), 4.52 (d, J=25.8Hz, 1H) , 4.34 (s, 1H), 3.25 (tt, J=7.3, 5.2Hz, 1H), 2.30 (s, 4H), 1.52 (p, J=5.3Hz, 4H), 1.42 (s, 2H);

[0300] 13 C NMR (151MHz, DMSO) δ 147.63, 145.04, 136.47, 120.80, 109.71, 100.89, 54.60, 54.60, 50.53, 50.53, 40.53, 25.56, 25.56, 24.30;

[0301] MS (+ESI) m / z: 341.2 (M+H) + ).

[0302] Example 38

[0303]

[0304] The preparation method of 7-(3-(4-fluoropiperidin-1-yl)azacyclobutane-1-yl)-4-nitro-1-oxobenzofurazan (compound A38) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1-(azacyclobutane-3-yl)-4-fluoropiperidine. A red powder (denoted as compound A38) was obtained with a yield of 36.7%.

[0305] Compound A38 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0306] 1 H NMR (600MHz, DMSO) δ 8.34 (d, J=9.0Hz, 1H), 5.89 (d, J=9.1Hz, 1H), 4.73 (dtd, J=48.5, 7.0, 3.6Hz, 2H), 4.51 (s, 1H ), 4.35 (s, 1H), 4.15 (s, 1H), 2.47 (s, 3H), 2.31 (s, 2H), 1.91-1.82 (m, 2H), 1.77-1.71 (m, 2H);

[0307] 13 C NMR (151MHz, DMSO) δ 147.62, 145.00, 136.49, 120.88, 109.73, 100.90, 54.04, 54.04, 45.90, 40.53, 31.28, 31.28, 31.15, 31.15;

[0308] MS (+ESI) m / z: 338.3 (M+H) + ).

[0309] Example 39

[0310]

[0311] The preparation method of 7-(1,4-diazabicyclo[3.2.2]nonane-4-yl)-4-nitro-1-oxobenzofuran (compound A39) was basically the same as that in Example 1, except that in step (4), N-ethylpiperazine was replaced by an equimolar amount of 1,4-diazabicyclo[3.2.2]nonane. A red powder (denoted as compound A39) was obtained with a yield of 22.8%.

[0312] Compound A39 1 H NMR, 13 The specific detection results of C NMR and MS (+ESI) are as follows:

[0313] 1 H NMR (600MHz, DMSO) δ 8.39 (d, J=9.0Hz, 1H), 6.46-6.44 (m, 1H), 3.77 (t, J=5.6Hz, 2H), 3.07-3.01 (m, 7H ), 2.16 (dddd, J=17.3, 10.8, 6.5, 3.7Hz, 2H), 1.97 (ddt, J=15.1, 10.1, 4.9Hz, 2H);

[0314] 13 C NMR (151MHz, DMSO) δ 148.72, 148.23, 136.56, 134.18, 111.88, 106.81, 46.05, 46.05, 45.94, 45.85, 25.27, 9.09, 9.09;

[0315] MS (+ESI) m / z: 306.2 (M+H) + ).

[0316] III. Tumor Suppressive Activity of Benzofuran Derivatives

[0317] 1. Inhibition rate of benzofuran derivatives on tumor cells

[0318] The inhibition rates of the above-mentioned benzofuran derivatives on human tumor cells HeLa (cervical cancer cells), A549 (lung cancer cells), HCT116 (colorectal cancer cells), and MKN45 (gastric cancer cells) were determined by the MTT assay, with XI-006 as a positive control. Lead compounds with tumor-inhibiting potential were screened by comparison.

[0319] (1) Preparation of drugs and reagents

[0320] Weigh a certain mass of the sample to be tested, prepare a stock solution with a concentration of 10 mM, and store it frozen at -20℃.

[0321] (2) Cell culture and passage

[0322] Adherent cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C in a 5% CO2 incubator. When the adherent cells reached 70%-90% confluence, the original medium was discarded, and the cells were washed twice with sterile PBS buffer. The cells were then digested with 1 mL of trypsin cell digestion solution to detach them from the culture dish. The cells were gently agitated with a pipette to completely detach them and form a single-cell suspension. The suspension was then displaced proportionally into new culture dishes for further culture.

[0323] (3) Drug treatment

[0324] Four different tumor cell lines in logarithmic growth phase (HCT116 colorectal cancer cells, HeLa cervical cancer cells, MKN45 gastric cancer cells, and A549 lung cancer cells) were adjusted to a cell concentration of 3000 cells / mL using DMEM complete medium. 100 μL of cell suspension was added to each well of a 96-well cell culture plate and cultured at 37°C in a 5% CO2 incubator for 24 h (cell adhesion). Then, 100 μL of medium containing 2 μM of different drugs was added to the drug wells, with 3 replicates for each drug. An equal volume (100 μL) of drug-free medium was added to the control wells, with 3 replicates. The cells were cultured at 37°C in a 5% CO2 incubator for 72 h. After culture, 20 μL of 5 mg / mL MTT solution was added to each well of a 96-well cell culture plate, and the plate was then incubated at 37°C in a 5% CO2 incubator for 2 hours. The liquid in the 96-well cell culture plate was then discarded, and 150 μL of dimethyl sulfoxide (DMSO) was added to each well. The 96-well cell culture plate was then shaken on a shaker until the purple crystals were completely dissolved. The absorbance of each well was measured at 570 nm using a microplate reader. The inhibition rate of each drug against tumor cells was calculated using the following formula:

[0325] Inhibition rate (%) = (1-A) 570给药孔 / A 570对照孔 ) × 100%

[0326] Calculations showed that at an administered concentration of 2 μM, the inhibition rates of compounds A1 to A39 and the positive control XI-006 against four tumor cell lines—HeLa, A549, HCT116, and MKN45—were as follows:

[0327] Table 1-1 Inhibition rates of various drugs on tumor cells HeLa, A549, HCT116, and MKN45 (%)

[0328]

[0329] Table 1-2 Inhibition rates (%) of various drugs on tumor cells HeLa, A549, HCT116, and MKN45

[0330]

[0331] From Table 1-1 and Table 1-2, we can see that:

[0332] (1) The benzofuran derivatives A3, A4, A11, A12, A13, A14, A15, A25 and A39 provided by the present invention can significantly inhibit the proliferation of tumor cells HeLa, and the inhibitory effect is significantly better than that of the original compound XI-006, showing good anti-tumor activity and having the potential to be developed into anti-cervical cancer drugs.

[0333] (2) The benzofuran derivatives A3, A4, A10, A11, A12, A13, A14, A15, A19, A20, A22, A23, A31, A32 and A39 provided by the present invention can significantly inhibit the proliferation ability of tumor cells A549, and the inhibitory effect is significantly better than that of the original compound XI-006, showing good anti-tumor activity and having the potential to be developed into anti-lung cancer drugs;

[0334] (3) The benzofuran derivatives A3A13 and A15 provided by the present invention can significantly inhibit the proliferation of tumor cells HCT116, and the inhibitory effect is significantly better than that of the original compound XI-006, showing good anti-tumor activity and having the potential to be developed into anti-colorectal cancer drugs.

[0335] (4) The benzofuran derivatives A3, A4, A11, A12, A13, A14, A15, A25 and A39 provided by the present invention can significantly inhibit the proliferation of tumor cells MKN45. The inhibitory effect is significantly better than that of the original compound XI-006, showing good anti-tumor activity and has the potential to be developed into anti-gastric cancer drugs.

[0336] 2. IC50 of the preferred compound on the inhibitory effect of tumor cell proliferation 50 value

[0337] The IC50 of the selected compounds against the proliferation of HeLa, A549, HCT116, and MKN45 tumor cells was determined using the MTT assay. 50 value.

[0338] (1) Preparation of drugs and reagents

[0339] Weigh a certain mass of the sample to be tested, prepare a stock solution with a concentration of 10 mM, and store it frozen at -20℃.

[0340] (2) Cell culture and passage

[0341] Adherent cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C in a 5% CO2 incubator. When the adherent cells reached 70%-90% confluence, the original medium was discarded, and the cells were washed twice with sterile PBS buffer. Then, the cells were digested with 1 mL of trypsin cell digestion solution to detach them from the culture dish. The cells were gently agitated with a pipette to completely detach them and form a single-cell suspension. The suspension was then displaced proportionally into new culture dishes for continued culture.

[0342] (3) Drug treatment

[0343] Four different tumor cell lines in logarithmic growth phase (HCT116 colorectal cancer cells, HeLa cervical cancer cells, MKN45 gastric cancer cells, and A549 lung cancer cells) were adjusted to a cell concentration of 3000 cells / mL using DMEM complete medium. 100 μL of cell suspension was added to each well of a 96-well cell culture plate and cultured at 37°C in a 5% CO2 incubator for 24 h (cell adhesion). Then, 100 μL of fresh medium containing different concentrations of the drug (7 concentration gradients for each drug: 8 μM, 4 μM, 2 μM, 1 μM, 0.5 μM, 0.25 μM, and 0.125 μM) was added to each well of the drug treatment plate. Each concentration was tested in triplicate. An equal volume (100 μL) of drug-free medium was added to each control well, also tested in triplicate. The plates were cultured at 37°C in a 5% CO2 incubator for 72 h. After culture, 20 μL of 5 mg / mL MTT solution was added to each 96-well cell culture plate, and the plate was then incubated at 37°C in a 5% CO2 incubator for 2 hours. The liquid in the 96-well cell culture plate was then discarded, and 150 μL of dimethyl sulfoxide (DMSO) was added to each well. The plate was then shaken on a shaker until the purple crystals were completely dissolved. The absorbance of each well was measured at 570 nm using a microplate reader. The inhibition rate of different drug concentrations on tumor cells was calculated using the aforementioned formula, and the IC50 of each drug on tumor cell proliferation inhibition was also calculated. 50 value.

[0344] IC50 of each drug against tumor cells HeLa, A549, HCT116, and MKN45 50The calculated values ​​are as follows:

[0345] Table 2. IC50 values ​​of the inhibitory effects of each drug on the proliferation of HeLa, A549, HCT116, and MKN45 tumor cells. 50 Value (μM)

[0346]

[0347] Table 2 shows the IC50 values ​​of compounds A3, A4, A11, A12, A13, A14, A15, A25, A31, A32, and A39 on the proliferation inhibition of four tumor cell lines: HeLa, A549, HCT116, and MKN45. 50 All of these compounds showed varying degrees of superiority over the original compound XI-006. Among them, compound A3 exhibited the strongest inhibitory effect on HeLa cells and HCT116 cells, being 10 times and 3.5 times better than XI-006, respectively. Compound A39 showed the strongest inhibitory effect on A549 cells and MKN45 cells, being 2.8 times and 10.8 times better than XI-006, respectively.

[0348] In summary, the benzofuran derivatives (compounds A3, A4, A11, A12, A13, A14, A15, A19, A20, A22, A23, A25, A31, A32 and A39) provided by this invention can significantly inhibit the proliferation of tumor cells HeLa, A549, HCT116 or MKN45, showing good anti-tumor activity and having the potential to be developed into anti-tumor drugs.

[0349] It should be noted that the above embodiments are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art can make other variations or modifications based on the above description. It is impossible to exhaustively list all possible implementations here. All obvious variations or modifications derived from the technical solutions of this invention are still within the scope of protection of this invention.

Claims

1. A benzofuran derivative, characterized in that, The structure of the benzofuran derivative is shown below: 。 2. The method for preparing the benzofuran derivative according to claim 1, characterized in that, Includes the following steps: (1) Compound I was prepared using 4-chloro-2-nitroaniline as a raw material; (2) Compound II was prepared using compound I as a raw material; (3) Compound III was prepared from compound II; (4) Compound III was dissolved in dichloromethane, and triethylamine and a substituted amine were added. The substituted amine was 4-diazabicyclo[3.2.2]nonane. The mixture was stirred at room temperature. After the reaction was completed, the product was purified and dried to obtain the benzofuran derivative. The structures of compounds I, II, and III are shown below: 、 、 。 3. The method for preparing the benzofuran derivative according to claim 2, characterized in that, In step (1), the method for preparing compound I is as follows: Sodium hydroxide was dissolved in anhydrous methanol under ice-water bath conditions, and 4-chloro-2-nitroaniline and a 5% sodium hypochlorite aqueous solution were added. The ratio of sodium hydroxide, anhydrous methanol, 4-chloro-2-nitroaniline and sodium hypochlorite aqueous solution was 30.5 mmol: 73 mL: 29.05 mmol: 87.15 mL. The mixture was stirred for 2 h under ice-water bath conditions. After the reaction was completed, the product was purified and dried to obtain compound I.

4. The method for preparing the benzofuran derivative according to claim 2, characterized in that, In step (2), the method for preparing compound II is as follows: Compound I was dissolved in concentrated sulfuric acid. Under ice-water bath conditions, a mixture of fuming nitric acid and concentrated sulfuric acid was slowly added dropwise. The ratio of compound I, concentrated sulfuric acid used to dissolve compound I, fuming nitric acid, and concentrated sulfuric acid mixed with fuming nitric acid was 22.6 mmol: 20 mL: 29.4 mmol: 3 mL. The mixture was stirred under ice-water bath conditions for 30 min. After the reaction was completed, the product was purified and dried to obtain compound II.

5. The method for preparing the benzofuran derivative according to claim 2, characterized in that, In step (3), the method for preparing compound III is as follows: Compound II was dissolved in glacial acetic acid at a ratio of 9 mmol: 14 mL. The mixture was heated under reflux for 3 h. After the reaction was completed, the product was purified and dried to obtain compound III.

6. The method for preparing the benzofuran derivative according to claim 2, characterized in that, In step (4), the molar ratio of compound III, triethylamine and substituted amine is 2.3:7:2.

8.

7. The application of a benzofuran derivative in the preparation of an anti-cervical cancer drug, characterized in that, The benzofuran derivative is: , , , , , , , and .

8. The application of a benzofuran derivative in the preparation of an anti-lung cancer drug, characterized in that, The benzofuran derivative is: 。 9. The application of a benzofuran derivative in the preparation of an anti-gastric cancer drug, characterized in that, The benzofuran derivative is: , , , , , , , and .

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