Anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of synthesis of compounds and drug application, and discloses a cinnamyl piperazine and 2-phenylbenzimidazole conjugate and a preparation method and application thereof, the structural general formula of the conjugate is as follows: # imgabs0 #. The cinnamyl piperazine and 2-phenylbenzimidazole conjugate and pharmaceutically acceptable salts thereof are synthesized for the first time, and the cinnamyl piperazine and 2-phenylbenzimidazole conjugate has the structural formula as shown in the specification. It is found for the first time that the compound has good anti-tumor activity, can significantly inhibit proliferation activity of liver cancer, triple negative breast cancer cells, ovarian cancer and cis-platinum drug-resistant ovarian cancer cells in vitro, has small toxicity to normal cells, and can inhibit migration ability of tumor cells. Therefore, the compound synthesized by the invention has a good anti-tumor application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of the synthesis and pharmaceutical application of compounds, and in particular to an antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate, and its preparation method and application. Background Art

[0002] In recent years, cinnamyl piperazine derivatives have been widely studied due to their extensive pharmacological applications. They have played a certain pharmacological role in central nervous system drugs, antitumor drugs, antiviral drugs, cardiovascular drugs, enzyme inhibitors, etc.

[0003] In 2010, Brossard et al. found that among a series of newly synthesized piperazinyl bile acid derivatives, the derivatives containing the cinnamyl piperazine fragment showed good antitumor activity, including a variety of human tumor cells (GBM, KMS-11, and HCT-116), and the IC 50 reached as low as 8.5 μM at best. Currently, the marketed drug flunarizine containing the cinnamyl piperazine fragment is mainly used to treat various nervous system diseases and migraines. In 2020, Chen et al. found that flunarizine inhibited the activation of the Akt pathway in human glioblastoma cells (U-87MG), and the combination of flunarizine and temozolomide could enhance cytotoxicity. Therefore, cinnamyl piperazine is expected to serve as a pharmacodynamic fragment in the research of antitumor drugs and play its role in inhibiting tumor activity.

[0004] In addition, Kim et al. found that 2-phenylbenzimidazole-5-sulfonic acid could inhibit the expression and secretion of vascular endothelial growth factor in ovarian cancer SKOV-3 cells, inhibit capillary-like tubular structures and angiogenesis in vitro, and down-regulate the expression and activity of matrix metalloproteinases in SKOV-3 cells. Liu et al. found that compounds containing 2-phenylbenzimidazole could induce apoptosis in human chondrosarcoma cell lines (JJ012 and SW1353). Therefore, 2-phenylbenzimidazole can be introduced as a pharmacophore into the design and synthesis of antitumor compounds.

[0005] The research and development of antitumor drugs has always been a hot topic in the research and development of new drugs. However, existing antitumor chemotherapy drugs have defects such as high toxicity, low selectivity for normal cells and tumor cells, and homogenization of generic drugs. The research motivation of the present invention is to discover a new antitumor compound that selectively kills tumor cells and has low toxicity to normal cells.

[0006] The research strategy of the present invention is to use the molecular hybridization method to combine two pharmacophores, cinnamyl piperazine and 2-phenylbenzimidazole, in one molecule to synthesize a novel cinnamyl piperazine and 2-phenylbenzimidazole conjugate. Through in vitro activity research, it is found that the conjugate has significantly higher antitumor activity than the two individual pharmacophores, and has good water solubility, showing the potential to become a new type of antitumor drug. Summary of the Invention

[0007] The object of the present invention is to overcome the deficiencies in the prior art and provide an antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate, its preparation method and application.

[0008] The technical solution adopted by the present invention to solve its technical problems is as follows:

[0009] An antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof, the general structural formula thereof is as follows:

[0010]

[0011] Wherein, on the linking bridge, X = oxygen, nitrogen, sulfur, carbon, n = 1 to 6; R 1 is hydrogen, methoxy, hydroxy, trifluoromethyl, fluorine, chlorine, cyano, formic acid, methyl formate, ethyl formate, methyl, ethyl, propyl, butyl, carbamoyl, methylcarbamoyl, cyclopropylcarbamoyl, propylcarbamoyl substituted at the 1-3 positions; R 2 is hydrogen, acetyl, methyl, ethyl, propyl, butyl, butyronitrile, propionitrile, acetonitrile; R 3 is hydrogen, methoxy, hydroxy, methyl, ethyl, propyl, butylmethylamino, dimethylamino substituted at the 1'-3' positions.

[0012] Furthermore, the structural formula of the antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof is one of the following:

[0013]

[0014] The general preparation method of the antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as described above includes the following steps:

[0015] First, cinnamyl bromide is used to connect N-tert-butoxycarbonyl (Boc) piperazine to obtain compound 15, and the Boc protecting group is removed to obtain compound 16; triethylene glycol and its analogs are brominated to obtain compound 17; p-hydroxybenzaldehyde reacts with 1,2-dibromoethane, 2,2'-dibromodiethyl ether, compound 17 and its analogs respectively to obtain the corresponding ethers 18-20; further connect compound 16 to obtain compounds 21-23 with different linking bridge lengths; finally, react with substituted o-phenylenediamine to obtain the end products 1-8, and the corresponding hydrochlorides can be obtained by introducing hydrogen chloride gas into a methanol or acetone solution. For example, introducing hydrogen chloride gas into compound 1 can obtain hydrochloride 1a.

[0016] Furthermore, its reaction route is as follows:

[0017]

[0018]

[0019] Alternatively, reacting 3,4-diaminobenzoic acid with cyclopropylamine gives 3,4-diamino-N-cyclopropylbenzamide (24), which reacts with compound 21 to give the final product 9, and its reaction route is as follows:

[0020]

[0021] Alternatively, the cyano group of compound 6 is hydrolyzed to give compound 10, and its reaction route is as follows:

[0022]

[0023] Compound 1 reacts with an alkyl halide under basic conditions to give imidazole nitrogen derivatized products 11-13; alternatively, compound 1 reacts with an acyl chloride to give amide 14, and its reaction route is as follows:

[0024]

[0025] Alternatively, a substituted cinnamylpiperazine intermediate 30 is prepared through a series of reactions, and then through similar steps as above, it reacts with compound 19 to give compound 31, and finally reacts with o-phenylenediamine to give compound 15, and its reaction route is as follows:

[0026]

[0027] Use of the antitumor cinnamylpiperazine and 2-phenylbenzimidazole conjugate as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating liver cancer.

[0028] Use of the antitumor cinnamylpiperazine and 2-phenylbenzimidazole conjugate as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating breast cancer and triple-negative breast cancer.

[0029] Use of the antitumor cinnamylpiperazine and 2-phenylbenzimidazole conjugate as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating ovarian cancer and drug-resistant ovarian cancer.

[0030] Use of the antitumor cinnamylpiperazine and 2-phenylbenzimidazole conjugate as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating tumors.

[0031] The advantages and positive effects achieved by the present invention are as follows:

[0032] 1. The cinnamyl piperazine and 2-phenylbenzimidazole conjugate of the present invention are synthesized for the first time, have good anti-tumor cell proliferation activity, and have low toxicity to normal human liver cells, which can make up for the defect of high toxicity of existing anti-tumor chemotherapy drugs and has outstanding innovation.

[0033] 2. In addition to having anti-hepatocellular carcinoma activity, the compounds of the present invention also have the activity of inhibiting the proliferation of triple-negative breast cancer, ovarian cancer and cisplatin-resistant ovarian cancer, indicating that the compounds have broad application prospects.

[0034] 3. The synthesis method of the compounds in the present invention is simple to operate, and the raw materials and reagents are cheap and easily available, which is suitable for large-scale production and development.

[0035] 4. Cisplatin-resistant ovarian cancer has always been a difficult problem in clinical treatment, with a high recurrence rate and low survival rate of patients. The compounds in the present invention make up for the shortage of such drugs;

[0036] 5. Some of the compounds in the present invention can be prepared into hydrochloride salts, which have good water solubility and high potential for drug formation.

[0037] 6. The present invention uses the method of molecular hybridization to combine two pharmacophores of cinnamyl piperazine and 2-phenylbenzimidazole in one molecule, synthesizes a novel cinnamyl piperazine and 2-phenylbenzimidazole conjugate, and through in vitro activity research, it is found that the conjugate has significantly higher anti-tumor activity than the two individual pharmacophores, and has good water solubility and the potential to become a new anti-tumor drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 1H NMR spectrum of Compound 1 in deuterated chloroform in the present invention;

[0039] Figure 2 1H NMR spectrum of Compound 2 in deuterated chloroform in the present invention;

[0040] Figure 3 1H NMR spectrum of Compound 3 in deuterated chloroform in the present invention;

[0041] Figure 4 1H NMR spectrum of Compound 4 in deuterated chloroform in the present invention;

[0042] Figure 5 1H NMR spectrum of Compound 5 in deuterated chloroform in the present invention;

[0043] Figure 6 1H NMR spectrum of Compound 6 in deuterated chloroform in the present invention;

[0044] Figure 7 1H NMR spectrum of Compound 7 in deuterated chloroform in the present invention;

[0045] Figure 8 1H NMR spectrum of compound 8 of the present invention in deuterochloroform;

[0046] Figure 9 1H NMR spectrum of compound 9 of the present invention in deuteromethanol;

[0047] Figure 10 1H NMR spectrum of compound 10 of the present invention in deuteromethanol;

[0048] Figure 11 1H NMR spectrum of compound 11 of the present invention in deuterochloroform;

[0049] Figure 12 1H NMR spectrum of compound 12 of the present invention in deuterochloroform;

[0050] Figure 13 1H NMR spectrum of compound 13 of the present invention in deuterochloroform;

[0051] Figure 14 1H NMR spectrum of compound 14 of the present invention in deuterochloroform;

[0052] Figure 15 1H NMR spectrum of compound 15 of the present invention in deuterochloroform;

[0053] Figure 16 1H NMR spectrum of compound 1a of the present invention in heavy water;

[0054] Figure 17 Cell number analysis chart of the effect of compound 1 of the present invention on the longitudinal migration ability (transwell) of cisplatin-resistant ovarian cancer (A2780 / DDP) cells. Compared with the solvent blank group (DMSO group), ***p < 0.001. Detailed implementation manners

[0055] The present invention will be further described below in conjunction with embodiments. The following embodiments are narrative and not restrictive, and the protection scope of the present invention cannot be limited by the following embodiments.

[0056] All kinds of experimental operations involved in the specific embodiments are conventional techniques in the art. For the parts not specifically noted in this article, those of ordinary skill in the art can refer to various commonly used reference books, scientific and technological literatures, or relevant specifications, manuals, etc. before the filing date of the present invention for implementation.

[0057] An antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof, the structural general formula is as follows:

[0058]

[0059] Among them, on the connecting bridge, X = oxygen, nitrogen, sulfur, carbon, and n = 1 to 6; R 1 is hydrogen, methoxy, hydroxy, trifluoromethyl, fluorine, chlorine, cyano, formic acid, methyl formate, ethyl formate, methyl, ethyl, propyl, butyl, carbamoyl, methylcarbamoyl, cyclopropylcarbamoyl, propylcarbamoyl substituted at the 1-3 positions; R 2 is hydrogen, acetyl, methyl, ethyl, propyl, butyl, butyronitrile, propionitrile, acetonitrile; R 3 is hydrogen, methoxy, hydroxy, methyl, ethyl, propyl, butylmethylamino, dimethylamino substituted at the 1'-3' positions.

[0060] Preferably, the structural formula of the anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or its pharmaceutically acceptable salt is one of the following:

[0061]

[0062] The general method for preparing the anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or its pharmaceutically acceptable salt as described above includes the following steps:

[0063] First, cinnamyl bromide is used to connect N-tert-butoxycarbonyl (Boc) piperazine to obtain compound 15, and the Boc protecting group is removed to obtain compound 16; triethylene glycol and its analogs are brominated to obtain compound 17; p-hydroxybenzaldehyde reacts with 1,2-dibromoethane, 2,2'-dibromodiethyl ether, compound 17 and its analogs respectively to obtain the corresponding ethers 18-20; further connect compound 16 to obtain compounds 21-23 with different connecting bridge lengths; finally, react with substituted o-phenylenediamine to obtain the end products 1-8, and the corresponding hydrochlorides can be obtained by introducing hydrogen chloride gas into a methanol or acetone solution. For example, introducing hydrogen chloride gas into compound 1 can obtain hydrochloride 1a.

[0064] Preferably, the reaction route is:

[0065]

[0066]

[0067] Alternatively, reacting 3,4-diaminobenzoic acid with cyclopropylamine gives 3,4-diamino-N-cyclopropylbenzamide (24), which reacts with compound 21 to obtain the end product 9, and the reaction route is:

[0068]

[0069] Alternatively, the cyano group of compound 6 is hydrolyzed to obtain compound 10, and the reaction route is as follows:

[0070]

[0071] Compound 1 reacts with an alkyl halide under basic conditions to obtain imidazole nitrogen derivatization products 11-13; alternatively, compound 1 reacts with an acyl chloride to obtain amide 14, and the reaction route is as follows:

[0072]

[0073] Alternatively, a substituted cinnamyl piperazine intermediate 30 is prepared through a series of reactions, and then through similar steps as above, it reacts with compound 19 to obtain compound 31, and finally reacts with o-phenylenediamine to obtain compound 15, and the reaction route is as follows:

[0074]

[0075] Use of the anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as described above in the preparation of a drug for treating liver cancer.

[0076] Use of the anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as described above in the preparation of a drug for treating breast cancer and triple-negative breast cancer.

[0077] Use of the anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as described above in the preparation of a drug for treating ovarian cancer and drug-resistant ovarian cancer.

[0078] Use of the anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as described above in the preparation of a drug for treating tumors.

[0079] Specifically, the related preparation and detection are as follows:

[0080] The synthetic routes of compounds 1-31 are as follows:

[0081] Among them, the synthetic routes of the final products 1-8 and hydrochloride 1a:

[0082]

[0083]

[0084] Example 1 Synthesis of 2-(4-(2-(4-cinnamylpiperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazole (1)

[0085] (1) Synthesis of Boc-cinnamyl piperazine (16)

[0086] To a 50 mL round-bottom flask, cinnamyl bromide (1.46 g, 7.41 mmol), 20 mL of acetonitrile, N-tert-butoxycarbonyl (Boc) piperazine (4.14 g, 22.23 mmol), and anhydrous sodium carbonate (2.36 g, 22.23 mmol) were successively added, and the mixture was stirred at room temperature for 5 h. After the reaction was complete, the reaction solution was directly evaporated to dryness. The obtained crude product was dissolved in 20 mL of ethyl acetate (EA) and 20 mL of water, and the aqueous layer was extracted with EA (20 mL × 3). The organic phases obtained from the three extractions were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4 dried, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography [petroleum ether (PE):EA = 10:1 - 1:1, V / V, volume ratio] to obtain 16 as a white solid product with a yield of 97.6%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 7.38 (d, J = 7.2 Hz, 2H), 7.31 (t, J = 7.6 Hz, 2H), 7.24 (t, J = 7.2 Hz, 1H), 6.54 (d, J = 16.0 Hz, 1H), 6.23 - 6.31 (m, 1H), 3.48 (s, 4H), 3.20 (d, J = 6.4 Hz, 2H), 2.48 (s, 4H), 1.46 (s, 9H).

[0087] (2) Synthesis of 1-cinnamylpiperazine (17)

[0088] Compound 16 (1.0 g, 3.31 mmol) was dissolved in 2.5 mL of anhydrous dichloromethane (DCM). At 0 °C, 0.5 mL of trifluoroacetic acid (TFA) was slowly added to the reaction solution, and the mixture was stirred at room temperature for 5 h. After the reaction was complete, the pH of the system was adjusted to 8 with Et 3 N, and the solvent and residual TFA were removed by concentration under reduced pressure to obtain the crude product of compound 17, which could be directly used for the next reaction without purification.

[0089] (3) Synthesis of 4-(2-bromomethoxy)benzaldehyde (19)

[0090] p-Hydroxybenzaldehyde (2 g, 16.38 mmol) was dissolved in 2.5 mL of anhydrous DMF. While stirring, 1,2-dibromoethane (14.12 mL, 163.8 mmol) and potassium carbonate (6.7 g, 49.14 mmol) were added, and the mixture was heated under reflux at 80 °C for 9 h. After the reaction was completed, 25 mL of ice water was added to the system to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4Dry, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (PE:EA = 150:1 - 50:1, V / V, volume ratio) to obtain 19 of the white solid product with a yield of 82.09%. 1 H-NMR(400MHz,CDCl 3 )δ9.90(s,1H),7.85(d,J=8.8Hz,2H),7.02(d,J=8.8Hz,2H),4.38(t,J=6.4Hz,2H),3.67(t,J=6.0Hz,2H).

[0091] (4) Synthesis of 4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)benzaldehyde (22)

[0092] Dissolve compound 17 (505.48 mg, 2.21 mmol) in 3 mL of anhydrous acetonitrile, add compound 19, potassium carbonate (1.2 g, 8.68 mmol), and potassium iodide (183.4 mg, 1.11 mmol) thereto while stirring, and heat under reflux at 80 °C for 3 h. After the reaction is complete, add 20 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry over anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM:MeOH = 200:1 - 30:1, V / V, volume ratio) to obtain 22 of the white solid product with a yield of 84.47%. 1 H-NMR(400MHz,CDCl 3 )δ9.88(s,1H),7.83(d,J=8.8Hz,2H),7.37(d,J=7.2Hz,2H),7.31(t,J=7.3Hz,2H),7.23(t,J=7.2Hz,1H),7.00(d,J=8.8Hz,2H),6.53(d,J=7.2Hz,1H),6.24 - 6.31(m,1H),4.18(t,J=6.0Hz,2H),3.17(d,J=6.8Hz,2H),2.86(t,J=6.0Hz,2H),2.58 - 2.66(m,8H).

[0093] (5) Synthesis of 2-(4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazole (i.e., compound 1)

[0094] Compound 22 (50 mg, 0.143 mmol) was dissolved in 0.2 mL of anhydrous acetonitrile. o-Phenylenediamine (15 mg, 0.143 mmol), 30% hydrogen peroxide by mass concentration (9.27 μL, 0.572 mmol), and ammonium cerium nitrate (19.59 mg, 0.036 mmol) were successively added, and the mixture was heated at 55 °C for 2 h. After the reaction was complete, 10 mL of ice water was added to the system to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4 dried, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain a yellowish-brown oil, 1, with a yield of 63.5%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 8.07 (t, J = 4.0 Hz, 2H), 7.54 (s, 2H), 7.32 (d, J = 7.6 Hz, 2H), 7.21 - 7.29 (m, 2H), 7.15 - 7.17 (m, 3H), 6.79 (d, J = 8.4 Hz, 2H), 6.47 (d, J = 16.0 Hz, 1H), 6.19 - 6.26 (m, 1H), 4.00 (s, 2H), 3.12 (d, J = 6.8 Hz, 2H), 2.53 - 2.76 (m, 10H). 13 13C-NMR (100 MHz, CDCl 3 ) δ 160.3, 152.0, 136.7, 133.6, 128.6, 128.3, 127.6, 126.4, 125.9, 122.6, 122.6, 115.0, 65.9, 60.9, 57.0, 53.4, 52.9. HRMS (ESI-TOF) m / z calcd. for C 28 28 30 H 4 3 + N Figure 1 O [M + H]

[0095] : 461.2312, found 4691.2312. The 1H NMR spectrum of compound 1 is shown in

[0096] It should be noted that there seems to be an error in the "found 4691.2312" in the original text, which might be a typo. It should probably be a value closer to the calculated one for a more reasonable result. Also, the "Na SO " in the text seems to be incomplete or incorrect in its presentation.Compound 22 (50 mg, 0.143 mmol) was dissolved in 0.2 mL of anhydrous acetonitrile. 4-Methoxyo-phenylenediamine (19.7 mg, 0.143 mmol), 30% hydrogen peroxide by mass concentration (9.27 μL, 0.572 mmol), and ammonium cerium(IV) nitrate (19.59 mg, 0.036 mmol) were added successively, and the mixture was heated at 55 °C for 5 h. After the reaction was complete, 10 mL of ice water was added to the system to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4 dried, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain a yellow oil 2 with a yield of 62.4%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 7.94 (d, J = 7.6 Hz, 2H), 7.47 (d, J = 8.4 Hz, 1H), 7.37 (d, J = 7.2 Hz, 2H), 7.30 (t, J = 7.2 Hz, 2H), 7.23 (t, J = 7.2 Hz, 1H), 7.06 (s, 1H), 6.92 (d, J = 8.4 Hz, 2H), 6.86 (d, J = 8.0 Hz, 1H), 6.53 (d, J = 16.0 Hz, 1H), 6.24 - 6.31 (m, 1H), 4.12 (t, J = 5.2 Hz, 2H), 3.82 (s, 3H), 3.20 (d, J = 6.4 Hz, 2H), 2.84 (t, J = 5.2 Hz, 2H), 2.63 - 2.69 (m, 8H). 13 13C-NMR (100 MHz, CDCl 3 ) δ 160.1, 156.5, 136.8, 136.8, 133.7, 133.6, 130.6, 128.7, 128.0, 127.7, 127.3, 126.5, 126.0, 125.9, 122.9, 115.2, 115.0, 114.9, 65.9, 61.0, 57.1, 55.9, 53.5, 53.0, 53.0. HRMS (ESI-TOF) m / z calcd. for C 29 29 32 H 4 3 2 [M + H] + : 491.2417, found 491.2438. The 1H NMR spectrum of compound 2 is shown in Figure 2 .

[0097] Synthesis of Example 3 2-(4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-5-fluoro-1H-benzo[d]imidazole (3)

[0098] Dissolve compound 22 (50 mg, 0.143 mmol) in 0.2 mL of anhydrous acetonitrile, and successively add 4-fluoro-1,2-phenylenediamine (18 mg, 0.143 mmol), 30% hydrogen peroxide by mass concentration (9.27 μL, 0.572 mmol), ammonium cerium(IV) nitrate (19.59 mg, 0.036 mmol), and heat at 55 °C for 5 h. After the reaction is complete, add 20 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry over anhydrous Na 2 SO 4 dry, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM:MeOH = 150:1 - 30:1, V / V, volume ratio) to obtain yellow oil 3 with a yield of 76.5%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 7.94 (d, J = 8.4 Hz, 2H), 7.45 (s, 1H), 7.35 (d, J = 7.2 Hz, 2H), 7.29 (t, J = 7.2 Hz, 2H), 7.19 - 7.25 (m, 2H), 6.89 - 6.97 (m, 3H), 6.50 (d, J = 15.6 Hz, 1H), 6.21 - 6.28 (m, 1H), 4.10 (s, 2H), 3.15 (d, J = 6.4 Hz, 2H), 2.82 (t, J = 5.6 Hz, 2H), 2.56 - 2.64 (m, 8H). 13 13C-NMR (100 MHz, CDCl 3 ) δ 160.9, 160.6, 158.5, 153.4, 136.9, 133.6, 128.7, 128.3, 127.7, 126.5, 126.2, 122.5, 115.2, 111.1, 110.8, 66.1, 61.1, 57.2, 53.6, 53.1. HRMS (ESI-TOF) m / z calcd. for C 28 24 29 H 4 16 + FN Figure 3 .

[0099] Synthesis of Example 4 2-(4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-5-(trifluoromethyl)-1H-benzo[d]imidazole (4)

[0100] Compound 22 (50 mg, 0.143 mmol) was dissolved in 0.2 mL of anhydrous acetonitrile. 4-(Trifluoromethyl)-1,2-benzenediamine (25.2 mg, 0.143 mmol), 30% hydrogen peroxide by mass concentration (9.27 μL, 0.572 mmol), and ammonium cerium(IV) nitrate (19.59 mg, 0.036 mmol) were successively added, and the mixture was heated at 55 °C for 5 h. After the reaction was complete, 10 mL of ice water was added to the system to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4 dried, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain a yellow oil, compound 4, with a yield of 86.38%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 7.93 (d, J = 8.0 Hz, 2H), 7.73 (s, 1H), 7.51 (d, J = 8.0 Hz, 1H), 7.21 - 7.26 (m, 3H), 7.11 - 7.18 (m, 3H), 6.64 (d, J = 8.0 Hz, 2H), 6.46 (d, J = 15.6 Hz, 1H), 6.03 - 6.10 (m, 1H), 4.57 (s, 1H), 3.83 (s, 2H), 3.29 (d, J = 5.2 Hz, 2H), 2.67 - 2.73 (m, 10H). 13 13C-NMR (100 MHz, MeOD) δ 162.2, 155.8, 138.8, 137.3, 129.7, 129.7, 129.4, 127.8, 126.1, 125.8, 125.5, 125.0, 123.0, 121.6, 120.4, 116.2, 66.5, 60.6, 57.4, 52.8, 52.8. HRMS (ESI-TOF) m / z calcd. for C 29 H 29 F 3 N 4 O [M+Na] + : 529.2186, found 529.2191. The 1H NMR spectrum of compound 4 is shown in Figure 4 .

[0101] Synthesis of Example 5 2-(4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-5-cyano-1H-benzo[d]imidazole (5)

[0102] Compound 22 (50 mg, 0.143 mmol) was dissolved in 0.2 mL of anhydrous acetonitrile. 3,4-Diaminobenzonitrile (19.04 mg, 0.143 mmol), 30% hydrogen peroxide by mass concentration (9.27 μL, 0.572 mmol), and ammonium cerium(IV) nitrate (19.59 mg, 0.036 mmol) were successively added, and the mixture was heated at 55 °C for 5 h. After the reaction was complete, 10 mL of ice water was added to the system to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The combined organic phases were washed with saturated brine and dried over anhydrous Na 2 SO 4 dried, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain a yellow oil 5 with a yield of 78%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 7.92 (d, J = 8.0 Hz, 2H), 7.71 (m, 1H), 7.46 (d, J = 8.0 Hz, 1H), 7.23 (s, 1H), 7.16 - 7.23 (m, 2H), 7.14 - 7.16 (m, 2H), 7.11 - 7.14 (m, 1H), 6.73 (d, J = 8.0 Hz, 2H), 6.45 (d, J = 15.6 Hz, 1H), 6.08 - 6.14 (m, 1H), 3.94 (s, 2H), 3.22 (d, J = 6.0 Hz, 2H), 2.66 - 2.73 (m, 10H). 13 13C-NMR (100 MHz, CDCl 3 ) δ 162.3, 156.4, 137.8, 137.5, 129.8, 129.7, 129.2, 127.6, 127.0, 122.8, 122.5, 120.8, 116.1, 106.1, 66.6, 60.9, 57.6, 53.1, 53.0. HRMS (ESI-TOF) m / z calcd. for C 29 24 29 H 5 3 + N Figure 5 3

[0103] O [M+K]

[0104] : 502.2004, found 502.2005. The 1H NMR spectrum of compound 5 is shown in Figure 5

[0103] Example 6 Synthesis of methyl 2-(4-(2-(4-cinnamylpiperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazole-5-carboxylate (6)

[0104] Compound 22 (50 mg, 0.143 mmol) was dissolved in 0.2 mL of anhydrous acetonitrile. Methyl 3,4-diaminobenzoate (23.76 mg, 0.143 mmol), 30% hydrogen peroxide by mass concentration (9.27 μL, 0.572 mmol), and ammonium cerium(IV) nitrate (19.59 mg, 0.036 mmol) were successively added, and the mixture was heated at 55 °C for 5 h. After the reaction was complete, 10 mL of ice water was added to the system to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4 dried, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 150:1 - 30:1, V / V, volume ratio) to obtain yellow oil 6 with a yield of 96.6%. 1 H-NMR (400 MHz, CDCl 3 ) δ 8.16 (s, 1H), 8.00 (d, J = 8.0 Hz, 2H), 7.79 (d, J = 8.4 Hz, 1H), 7.43 (s, 1H), 7.22 (d, J = 7.2 Hz, 2H), 7.16 (t, J = 6.8 Hz, 2H), 7.09 (t, J = 6.8 Hz, 1H), 6.73 (d, J = 8.4 Hz, 2H), 6.38 (d, J = 15.6 Hz, 1H), 6.09 - 6.16 (m, 1H), 3.93 (s, 2H), 3.75 (s, 3H), 3.03 (d, J = 6.4 Hz, 2H), 2.67 (s, 2H), 2.51 (s, 8H). 13 C-NMR (100 MHz, CDCl 3 ) δ 167.9, 160.6, 154.9, 136.6, 133.5, 128.7, 128.5, 127.5, 126.3, 125.8, 124.1, 122.0, 115.0, 65.8, 60.8, 56.9, 53.4, 52.9, 52.1. HRMS (ESI-TOF) m / z calcd. for C 30 H 32 N 4 O 3 [M+Na] + : 519.2367, found 519.2365. The 1H NMR spectrum of compound 6 is shown in Figure 6 .

[0105] Synthesis of Example 7 2-(4-(2-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)ethoxy)phenyl)-1H-benzo[d]imidazole (7)

[0106] (1) Synthesis of 4-(2-bromoethoxy)ethoxybenzaldehyde (20)

[0107] Dissolve p-hydroxybenzaldehyde (100 mg, 0.82 mmol) in 0.5 mL of anhydrous DMF. While stirring, add 2,2'-dibromo diethyl ether (1.05 mL, 8.2 mmol) and potassium carbonate (453 mg, 3.28 mmol), and heat under reflux at 80 °C for 4 h. After the reaction is completed, add 10 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry over anhydrous Na 2 SO 4 2SO4. Filter, and concentrate the filtrate under reduced pressure. Purify the crude product by silica gel column chromatography (DCM:MeOH = 200:1 - 150:1, V / V, volume ratio) to obtain the white solid product 20 with a yield of 65.3%.

[0108] (2) Synthesis of 4-(2-(4-cinnamylpiperazin-1-yl)ethoxy)ethoxy)benzaldehyde (23)

[0109] Dissolve compound 17 (88.3 mg, 0.435 mmol) in 1 mL of anhydrous acetonitrile. While stirring, add compound 20, potassium carbonate (120 mg, 0.87 mmol), and potassium iodide (17.69 mg, 0.145 mmol), and heat under reflux at 80 °C for 8 h. After the reaction is complete, add 10 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry over anhydrous Na 2 SO 4 2SO4. Filter, and concentrate the filtrate under reduced pressure. Purify the crude product by silica gel column chromatography (DCM:MeOH = 200:1 - 100:1, V / V, volume ratio) to obtain the white solid product 23 with a yield of 60.2%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 9.87 (s, 1H), 7.82 (d, J = 8.8 Hz, 2H), 7.38 (d, J = 7.6 Hz, 2H), 7.32 (t, J = 7.2 Hz, 2H), 7.23 - 7.27 (m, 1H), 7.01 (d, J = 8.4 Hz, 2H), 6.58 (d, J = 16 Hz, 1H), 6.27 - 6.34 (m, 1H), 4.21 (t, J = 4.4 Hz, 2H), 3.85 (t, J = 4.4 Hz, 2H), 3.79 (t, J = 5.6 Hz, 2H), 3.31 (d, J = 6.8 Hz, 2H), 2.78 - 2.83 (m, 10H).

[0110] (3) Synthesis of 2-(4-(2-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)ethoxy)phenyl)-1H-benzo[d]imidazole (7)

[0111] Dissolve compound 23 (51.9 mg, 0.13 mmol) in 0.2 mL of anhydrous acetonitrile. Sequentially add o-phenylenediamine (45.6 mg, 0.13 mmol), 30% hydrogen peroxide by mass concentration (8 μL, 0.52 mmol), ammonium cerium(IV) nitrate (35.6 mg, 0.065 mmol), and heat at 55 °C for 5 h. After the reaction is complete, add 10 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry over anhydrous Na 2 SO 4 dry, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM:MeOH = 150:1 - 30:1, V / V, volume ratio) to obtain yellow oil 7 with a yield of 31.7%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 8.05 (d, J = 8.8 Hz, 2H), 7.61 - 7.64 (m, 2H), 7.32 - 7.32 (m, 1H), 7.29 - 7.30 (m, 2H), 7.27 (s, 1H), 7.22 - 7.25 (m, 1H), 7.18 - 7.21 (m, 2H), 6.87 (d, J = 9.2 Hz, 2H), 6.48 (d, J = 15.8 Hz, 1H), 6.18 - 6.26 (m, 1H), 4.07 (t, J = 4.2 Hz, 2H), 3.73 (t, J = 4.5 Hz, 2H), 3.68 (t, J = 5.2 Hz, 2H), 3.20 (d, J = 6.8 Hz, 2H), 2.65 - 2.70 (m, 10H). 13 13C-NMR (100 MHz, CDCl 3 ) δ 160.2, 151.9, 139.1, 136.1, 135.5, 128.7, 128.5, 128.1, 126.6, 123.1, 122.6, 115.1, 69.7, 68.6, 67.4, 60.3, 57.4, 52.4, 51.9. HRMS (ESI-TOF) m / z calcd. for C 30 29 34 H 4 33 2 [M + H] + : 483.2755, found 483.2763. The 1H NMR spectrum of compound 7 is shown in Figure 7 .

[0112] Synthesis of 82-(4-(2-(2-(4-cinnamylpiperazin-1-yl)ethoxy)ethoxy)ethoxy)phenyl)-1H-benzo[d]imidazole (8)

[0113] (1) Synthesis of 1,2-bis(2-bromoethoxy)ethane (18)

[0114] Dissolve triethylene glycol (100 mg, 0.95 mmol) in 3 mL of anhydrous diethyl ether, and dropwise add PBr 3 (268 μL, 2.85 mmol) under an ice bath, and react at room temperature for 2 h. After the reaction is completed, add 10 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry with anhydrous Na 2 SO 4 Dry, filter, and concentrate the filtrate under reduced pressure to obtain the crude product as a white oily substance 18, with a yield of 69.1%.

[0115] (2) Synthesis of 4-(2-(2-(2-bromoethoxy)ethoxy)benzaldehyde (21)

[0116] Dissolve p-hydroxybenzaldehyde (63 mg, 0.5 mmol) in 0.5 mL of anhydrous DMF, and add compound 17 (600 mg, 2.5 mmol), potassium carbonate (285 mg, 2 mmol) while stirring, and heat under reflux at 80 °C for 3 h. After the reaction is completed, add 10 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry with anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM:MeOH = 200:1 - 150:1, V / V, volume ratio) to obtain the white solid product 21, with a yield of 93.2%.

[0117] (3) 4-(-(2-(4-cinnamylpiperazin-1-yl)ethoxy)ethoxy)ethoxybenzaldehyde (24)

[0118] Dissolve compound 17 (82 mg, 0.40 mmol) in 1 mL of anhydrous acetonitrile, and add compound 21 (76 mg, 0.27 mmol), potassium carbonate (111.9 mg, 0.81 mmol), potassium iodide (22.4 mg, 0.135 mmol) while stirring, and heat under reflux at 80 °C for 6 h. After the reaction is complete, add 10 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry with anhydrous Na 2 SO 4Dry, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM:MeOH = 200:1 - 100:1, V / V, volume ratio) to obtain 24 of the white solid product with a yield of 86.9%. 1 H-NMR(400MHz,CDCl 3 )δ9.87(s,1H),7.81(d,J=8.8Hz,2H),7.39-7.39(m,2H),7.31-7.37(m,2H),7.27-7.29(m,1H),7.01(d,J=8.7Hz,2H),6.60(d,J=15.8Hz,1H),6.18-6.26(m,1H),4.21(t,J=4.5Hz,2H),3.87(t,J=4.8Hz,2H),3.69-3.75(m,4H),3.63-3.66(m,2H),3.40(s,2H),2.90-2.99(m,10H).

[0119] (4) Synthesis of 2-(4-(2-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)ethoxy)ethoxy)phenyl)-1H-benzo[d]imidazole (8)

[0120] Dissolve compound 24 (60 mg, 0.15 mmol) in 0.2 mL of anhydrous acetonitrile, and successively add o-phenylenediamine (52.9 mg, 0.15 mmol), 30% hydrogen peroxide by mass concentration (10 μL, 0.6 mmol), ammonium cerium nitrate (41.1 mg, 0.075 mmol), and heat at 55 °C for 5 h. After the reaction is complete, add 20 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry with anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM:MeOH = 150:1 - 20:1, V / V, volume ratio) to obtain 8 of the yellow oil with a yield of 30.2%. 1 H-NMR(400MHz,CDCl 3 )δ8.06(d,J=8.7Hz,2H),7.63-7.66(m,2H),7.28-7.31(m,4H),7.20-7.25(m,3H),6.92(d,J=8.7Hz,2H),6.44(d,J=15.8Hz,1H),6.14-6.22(m,1H),4.12(t,J=4.2Hz,2H),3.81(t,J=4.5Hz,2H),3.60-3.3.69(m,6H),3.13(d,J=6.9Hz,2H),2.61-2.68(m,10H).13 C-NMR(100MHz,CDCl 3 )δ160.4,151.9,136.2,135.5,128.7,128.6,128.1,126.6,123.0,122.9,122.5,115.1,70.9,70.5,69.8,68.4,67.6,60.4,57.4,52.5,51.9.HRMS(ESI-TOF)m / z calcd.for C 32 H 38 N 4 O 3 [M+Na] + :549.2836,found 549.2843. The 1H NMR spectrum of compound 8 is shown in Figure 8 。

[0121] Synthesis of Example 9 2-(4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazole hydrochloride (1a)

[0122] Compound 1 (100 mg) was dissolved in 2 mL of acetone, and hydrochloric acid acetone solution (100 μg / mL, 0.5 mL) was added. The mixture was stirred at room temperature for 2 h, and a solid slowly precipitated. Stirring was stopped to obtain white solid 1a with a yield of 100%. 1 H-NMR(400MHz,D 2 O)δ7.93(d,J = 7.6Hz,2H),7.68(s,2H),7.53 - 5.54(m,4H),7.38 - 7.44(m,3H),7.22(d,J = 8.8Hz,2H),6.97(d,J = 16.0Hz,1H),6.28 - 6.35(m,1H),4.46(s,2H),4.09(d,J = 7.6Hz,2H),3.76(s,10H). The 1H NMR spectrum of compound 1a is shown in Figure 16 。

[0123] Among them, the synthetic route of the final product 9:

[0124]

[0125] Synthesis of Example 10 2-(4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-N-cyclopropyl-1H-benzo[d]imidazole-5-carboxamide (9)

[0126] (1) Synthesis of 3,4-Diamino-N-cyclopropylbenzamide (25)

[0127] Dissolve 3,4-diaminobenzoic acid (96 mg, 0.5 mmol) in anhydrous DMF (1 mL). While stirring, add cyclopropylamine (143 mg, 2.5 mmol), N,N'-diisopropylcarbodiimide (DIC, 315 mg, 2.5 mmol), and 4-dimethylaminopyridine (DMAP, 25 mg, 0.2 mmol) successively. React at room temperature for 3 h. After the reaction is complete, add 20 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry with anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure, and separate and purify the crude product by silica gel column chromatography (DCM:MeOH = 150:1 - 50:1, V / V, volume ratio) to obtain a transparent oil 25 with a yield of 48.39%.

[0128] (2) Synthesis of 2-(4-(2-(4-cinnamylpiperazin-1-yl)ethoxy)phenyl)-N-cyclopropyl-1H-benzo[d]imidazole-5-carboxamide (9)

[0129] Dissolve compound 22 (50 mg, 0.143 mmol) in 0.2 mL of anhydrous acetonitrile. Add 3,4-diamino-N-cyclopropylbenzamide (i.e., compound 25) (27.33 mg, 0.143 mmol), 30% hydrogen peroxide by mass concentration (9.27 μL, 0.572 mmol), and ammonium cerium nitrate (19.59 mg, 0.036 mmol) successively. Heat at 55 °C for 5 h. After the reaction is complete, add 10 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry with anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure, and separate and purify the crude product by silica gel column chromatography (DCM:MeOH = 150:1 - 30:1, V / V, volume ratio) to obtain a yellow solid 9 with a yield of 12.8%. 1H-NMR(400MHz,MeOD)δ8.05(d,J=8.8Hz,3H),7.71(d,J=8.0Hz,1H),7.60(t,J=9.6Hz,1H),7.40(d,J=7.2Hz,2H),7.30(t,J=7.2Hz,2H),7.22(t,J=7.2Hz,1H),7.12(d,J=8.8Hz,2H),6.61(d,J=16.0Hz,1H),6.24-6.31(m,1H),4.23(t,J=5.2Hz,2H),3.24(d,J=6.8Hz,2H),2.86-2.90(m,3H),2.70(s,8H),0.83(d,J=5.2Hz,2H),0.67(t,J=2.4Hz,2H). 13 C-NMR(100MHz,MeOD)δ172.3,162.3,138.0,135.9,130.0,129.7,128.8,127.4,125.4,123.2,116.2,66.7,61.6,58.0,54.0,53.5,24.1,6.6.HRMS(ESI-TOF)m / z calcd.for C 32 H 35 N 5 O 2 [M+H] + :522.2864,found 522.2873. The 1H NMR spectrum of compound 9 is shown in Figure 9 .

[0130] Among them, the synthetic route of the final product 10:

[0131]

[0132] Example 11 Synthesis of 2-(4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazole-5-carboxylic acid (10)

[0133] Dissolve compound 6 (see Example 6) (45 mg, 0.087 mmol) in an aqueous methanol solution with a volume concentration of 50%. While stirring, add lithium hydroxide (36.9 mg, 0.348 mmol). Heat at 55 °C for 24 h. After the reaction is completed, add formic acid to the system to adjust the pH to 3-4, then extract with diethyl ether (10 mL × 3), wash with saturated brine, and dry over anhydrous Na 2 SO 4 Dry, filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM:MeOH = 100:1 - 10:1, V / V, volume ratio) to obtain yellow solid 10 with a yield of 40.1%.1 1H-NMR(400MHz,MeOD)δ8.24(s,1H),8.05(d,J=8.2Hz,2H),7.93(d,J=8.4Hz,1H),7.55(d,J=8.4Hz,1H),7.42(d,J=7.5Hz,2H),7.31(t,J=7.4Hz,2H),7.23(t,J=7.2Hz,1H),7.11(d,J=8.2Hz,2H),6.66(d,J=15.8Hz,1H),6.26-6.33(m,1H),4.23(t,J=4.6Hz,2H),3.38(s,2H),2.80-2.94(m,10H). 13 13C-NMR(100MHz,MeOD)δ161.8,154.7,137.6,136.7,129.5,129.4,128.8,127.4,125.2,123.9,123.3,116.0,66.5,61.0,57.6,53.3,53.0.HRMS(ESI-TOF)m / z calcd.for C 29 H 30 N 4 O 3 [M+H] + :483.2391,found 483.2397. The 1H-NMR spectrum of compound 10 is shown in Figure 10 .

[0134] Among them, the synthetic routes of the final products 11 - 14 are as follows:

[0135]

[0136] Example 12 Synthesis of 4-(2-(4-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazol-1-yl)butanenitrile (11)

[0137] Dissolve compound 1 (25 mg, 0.057 mmol) in anhydrous DMF (1 mL), and successively add 4-chlorobutyronitrile (6 μL, 0.063 mmol) and potassium hydroxide (6.4 mg, 0.171 mmol) while stirring. React at 80 °C for 36 h. After the reaction is complete, add 20 mL of ice water to the system to quench the reaction, and extract the aqueous layer with EA (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry over anhydrous Na 2 SO 4 4. Filter, concentrate the filtrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain yellow oil 11 with a yield of 42.8%. 1H-NMR(400MHz,CDCl 3 )δ7.80(m,1H),7.62(d,J=8.8Hz,2H),7.37-7.42(m,3H),7.29-7.35(m,4H),7.21-7.25(m,1H),7.05(d,J=8.8Hz,2H),6.54(d,J=15.9Hz,1H),6.25-6.33(m,1H),4.42(t,J=7.2Hz,2H),4.19(t,J=5.7Hz,2H),3.21(d,J=6.7Hz,2H),2.89(t,J=5.7Hz,2H),2.64-2.71(m,8H),2.22(d,J=7.5Hz,2H),2.06-2.13(m,2H). 13 C-NMR(100MHz,CDCl 3 )δ160.3,153.5,143.2,137.0,135.4,133.4,130.7,128.7,127.6,126.4,123.1,122.8,122.5,120.2,118.3,115.2,109.6,66.2,61.1,57.2,53.7,53.2,43.0,29.8,25.7,14.8.HRMS(ESI-TOF)m / z calcd.for C 34 H 37 N 5 O 3 [M+H] + :564.2969,found 564.2975. The 1H-NMR spectrum of Compound 11 is shown in Figure 11 .

[0138] Synthesis of Example 13 2-(4-(2-(4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-1-methyl-1H-benzo[d]imidazole (12)

[0139] Compound 1 (50 mg, 0.114 mmol) was dissolved in anhydrous DMF (1 mL). While stirring, methyl iodide (14 μL, 0.228 mmol) and potassium hydroxide (30 mg, 0.342 mmol) were added successively. The reaction was carried out at 80 °C for 2 h. After the reaction was complete, 10 mL of ice water was added to the system to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The combined organic phases were washed with saturated brine and dried over anhydrous Na 2 SO 4 . The filtrate was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain yellow oil 12 with a yield of 39%.1 H-NMR (400 MHz, CDCl 3 ) δ 7.78 (m, 1H), 7.70 (d, J = 8.7 Hz, 2H), 7.37 - 7.39 (m, 3H), 7.28 - 7.33 (m, 4H), 7.21 - 7.25 (m, 1H), 7.05 (d, J = 8.7 Hz, 2H), 6.54 (d, J = 15.8 Hz, 1H), 6.25 - 6.33 (m, 1H), 4.19 (t, J = 5.7 Hz, 2H), 3.86 (s, 3H), 3.21 (d, J = 6.5 Hz, 2H), 2.89 (t, J = 5.7 Hz, 2H), 2.65 - 2.70 (m, 8H). 13 C-NMR (100 MHz, CDCl 3 ) δ 160.0, 153.8, 143.0, 136.8, 136.7, 130.9, 128.7, 127.7, 126.4, 122.8, 122.6, 122.4, 119.7, 114.8, 109.6, 66.1, 61.0, 57.1, 53.5, 53.1, 31.8. HRMS (ESI-TOF) m / z calcd. for C 29 H 32 N 4 O [M + H] + : 453.2649, found 453.2658. The Figure 12 .

[0140] Synthesis of 1-butyl-2-(4-(2-(4-cinnamylpiperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazole (13) in Example 14

[0141] Compound 1 (50 mg, 0.114 mmol) was dissolved in anhydrous DMF (1 mL). While stirring, bromobutane (15 μL, 0.140 mmol) and potassium hydroxide (12.8 mg, 0.228 mmol) were added successively. The reaction was carried out at 80 °C for 2 h. After the reaction was complete, 10 mL of ice water was added to the system to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous Na 2 SO 4 , filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain a yellow oil 13 with a yield of 38.8%. 1 H-NMR (400 MHz, CDCl 3)δ 7.80 (t, J = 5.5 Hz, 1H), 7.64 (d, J = 8.5 Hz, 2H), 7.38 (d, J = 7.3 Hz, 3H), 7.27 - 7.32 (m, 4H), 7.21 - 7.24 (m, 1H), 7.03 (d, J = 8.5 Hz, 2H), 6.54 (d, J = 15.8 Hz, 1H), 6.26 - 6.33 (m, 1H), 4.17 - 4.22 (m, 4H), 3.20 (d, J = 6.6 Hz, 2H), 2.89 (t, J = 5.5 Hz, 2H), 2.62 - 2.70 (m, 8H), 1.79 (t, J = 7.4 Hz, 2H), 1.27 (t, J = 7.4 Hz, 2H), 0.87 (t, J = 7.3 Hz, 3H). 13 C-NMR (100 MHz, CDCl 3 ) δ 159.9, 153.7, 143.2, 136.8, 135.7, 133.6, 130.8, 128.7, 127.7, 126.4, 123.2, 122.5, 122.3, 119.8, 114.8, 110.1, 66.1, 61.0, 57.1, 53.5, 53.1, 44.6, 31.9, 20.0, 13.7. HRMS (ESI-TOF) m / z calcd. for C 32 H 38 N 4 O [M + H] + : 495.3118, found 495.3123. The 1H-NMR spectrum of Compound 13 is shown in Figure 13 .

[0142] Example 15 Synthesis of 1-(2-(4-(2-(-4-Cinnamylpiperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazol-1-yl)ethan-1-one (14)

[0143] Compound 1 (30 mg, 0.068 mmol) was dissolved in anhydrous DCM (1 mL). Triethylamine (0.1 ml) and acetyl chloride (20 μl, 0.102 mmol) were successively added dropwise with stirring at 0 °C. The reaction was carried out at room temperature for 16 h. After the reaction was complete, 10 mL of ice water was added to quench the reaction, and the aqueous layer was extracted with EA (10 mL × 3). The organic phases were combined, washed with saturated brine, dried over anhydrous Na 2 SO 4 and filtered. The filtrate was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain a yellow oil 14 with a yield of 62.0%. 1 1H-NMR (400 MHz, CDCl3 ) δ 8.13 - 8.15 (m, 1H), 7.76 - 7.78 (m, 1H), 7.59 (d, J = 8.5 Hz, 2H), 7.37 - 7.41 (m, 4H), 7.30 (t, J = 7.4 Hz, 2H), 7.21 - 7.24 (m, 1H), 7.04 (d, J = 8.6 Hz, 2H), 6.54 (d, J = 8.6 Hz, 1H), 6.25 - 6.32 (m, 1H), 4.19 (t, J = 5.6 Hz, 2H), 3.19 (d, J = 6.2 Hz, 2H), 2.88 (t, J = 5.6 Hz, 2H), 2.60 - 2.68 (m, 8H), 2.22 (s, 3H). 13 C - NMR (100 MHz, CDCl 3 ) δ 170.5, 160.7, 142.5, 134.1, 130.9, 128.6, 127.6, 126.4, 125.4, 125.0, 124.1, 120.0, 115.1, 115.1, 66.2, 61.1, 57.1, 53.7, 53.1, 27.5. The 1H - NMR spectrum of compound 14 is shown in Figure 14 .

[0144] Among them, for the final product 15:

[0145]

[0146] Synthesis of Example 16 (E) - 2 - (4 - (2 - (4 - (3 - methoxyphenyl) allyl) piperazin - 1 - yl) ethoxy) phenyl) - 1H - benzimidazole (15)

[0147] (1) Synthesis of methyl (E) - 3 - (3 - methoxyphenyl) acrylate (26)

[0148] Dissolve 3 - methoxycinnamic acid (50 mg, 0.28 mmol) in 1 mL of anhydrous methanol. Dropwise add 0.1 mL of thionyl chloride at 0 °C and react at room temperature for 30 min. After the reaction is complete, add 2 mL of ice - water to quench the reaction. Extract the aqueous layer with DCM (10 mL × 3). Combine the organic phases, wash with saturated brine, and dry over anhydrous Na 2 SO 4 Dry, filter, and concentrate the filtrate under reduced pressure to obtain a white solid 26 with a yield of 92.6%. 1 1H - NMR (400 MHz, CDCl 3)δ 7.59 (d, J = 16 Hz, 1H), 7.21 (t, J = 8.1 Hz, 1H), 7.02 (d, J = 7.6 Hz, 1H), 6.95 (t, J = 2.1 Hz, 1H), 6.83 - 6.86 (m, 1H), 6.34 (d, J = 16.0 Hz, 1H), 3.72 (d, J = 6.8 Hz, 6H)

[0149] (2) Synthesis of (E)-3-(3-methoxyphenyl)prop-2-en-1-ol (27)

[0150] Compound 26 (50 mg, 0.26 mmol) was dissolved in 1.5 mL of anhydrous DCM. Under anhydrous and anaerobic conditions, 0.78 mL of diisobutylaluminum hydride (Dibal-H) was added dropwise to the system at -30 °C. After reacting for 40 min, when the reaction was complete, 0.4 mL of methanol and 0.4 mL of water were added to the system, and the aqueous phase was extracted with DCM (10 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4 After drying and filtration, the filtrate was concentrated under reduced pressure to obtain a white oil, 27, with a yield of 81.2%. 1 1H-NMR (400 MHz, CDCl 3 ) δ 7.14 (t, J = 7.9 Hz, 1H), 6.88 (d, J = 7.6 Hz, 1H), 6.83 (s, 1H), 6.71 (q, J 1 = 8.2 Hz, J 2 = 2.2 Hz, 1H), 6.48 (d, J = 15.9 Hz, 1H), 6.22 - 6.29 (m, 1H), 4.21 (d, J = 5.0 Hz, 2H), 3.71 (s, 3H).

[0151] (3) Synthesis of (E)-1-(3-bromoprop-1-en-1-yl)-3-methoxybenzene (28)

[0152] Compound 27 (50 mg, 0.305 mmol) was dissolved in 1.5 mL of anhydrous diethyl ether. Under argon protection and an ice bath, 53.6 μL of phosphorus tribromide was added dropwise. The reaction was carried out at room temperature for 40 min. After the reaction was complete, 5 mL of ice water was added to the system to quench the reaction, and the aqueous phase was extracted with DCM (10 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4 After drying and filtration, the filtrate was concentrated under reduced pressure to obtain a yellow oil, 28, with a yield of 80.6%.

[0153] (4) Synthesis of (E)-4-(3-(3-methoxyphenyl)allyl)piperazine-1-carboxylic acid tert-butyl ester (29)

[0154] Compound 28 (35 mg, 0.145 mmol) was dissolved in 1 mL of acetonitrile. While stirring, N-tert-butoxycarbonyl (Boc) piperazine (29.7 mg, 0.16 mmol) and potassium carbonate (40 mg, 0.29 mmol) were added successively. The reaction was carried out at room temperature for 5 h. After the reaction was complete, 5 mL of ice water was added to the system to quench the reaction. The aqueous phase was extracted with DCM (10 mL × 3). The organic phases were combined, washed with saturated brine, and dried over anhydrous Na 2 SO 4 2SO4, filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (DCM:MeOH = 100:1 - 20:1, V / V, volume ratio) to obtain white solid 29 with a yield of 82.2%. 1 1H-NMR (400 MHz, CDCl 3 3) δ 7.15 (t, J = 8 Hz, 1H), 6.89 (d, J = 7.6 Hz, 1H), 6.85 (s, 1H), 6.72 (q, J 1 = 8.2 Hz, J 2 = 2.1 Hz, 1H), 6.42 (d, J = 15.8 Hz, 1H), 6.14 - 6.22 (m, 1H), 3.74 (s, 3H), 3.39 (t, J = 4.6 Hz, 4H), 3.09 (d, J = 6.7 Hz, 2H), 2.37 (s, 4H), 1.39 (s, 9H).

[0155] (5) Synthesis of (E)-1-(3-(3-methoxyphenyl)allyl)piperazine (30)

[0156] Compound 29 (97 mg, 0.29 mmol) was dissolved in 1 mL of anhydrous dichloromethane. At 0 °C, 0.2 mL of trifluoroacetic acid was slowly added to the reaction solution, and the mixture was stirred at room temperature for 5 h. After the reaction was complete, the pH of the system was adjusted to 8 with Et 3 3N, and the solvent and residual TFA were removed by concentration under reduced pressure to obtain the crude product of compound 30, which could be directly used in the next reaction without purification.

[0157] (6) Synthesis of (E)-4-(2-(4-(3-methoxyphenyl)allyl)piperazin-1-yl)ethoxy)benzaldehyde (31)

[0158] Compound 30 (50 mg, 0.216 mmol) was dissolved in 1 mL of anhydrous acetonitrile. While stirring, compound 19 (103 mg, 0.45 mmol), potassium carbonate (186.6 mg, 1.35 mmol), and potassium iodide (37 mg, 0.225 mmol) were added successively. The reaction was carried out at 80 °C for 14 h. After the reaction was complete, the work-up method was the same as that for compound 22 to obtain compound 31 with a yield of 86.3%.1 H-NMR (400 MHz, CDCl 3 ) δ 9.87 (s, 1H), 7.82 (d, J = 8.7 Hz, 2H), 7.22 (t, J = 7.9 Hz, 1H), 6.96 - 7.01 (m, 3H), 6.92 (s, 1H), 6.78 - 6.81 (m, 1H), 6.52 (d, J = 15.8 Hz, 1H), 6.26 - 6.33 (m, 1H), 4.19 (t, J = 5.6 Hz, 2H), 3.80 (s, 3H), 3.22 (d, J = 6.4 Hz, 2H), 2.88 (t, J = 5.6 Hz, 2H), 2.66 - 2.71 (m, 8H).

[0159] (7) Synthesis of (E)-2-(4-(2-(4-(3-methoxyphenyl)allyl)piperazin-1-yl)ethoxy)phenyl)-1H-benzo[d]imidazole (15)

[0160] Dissolve compound 31 (95 mg, 0.22 mmol) in 0.2 mL of anhydrous acetonitrile, and successively add o-phenylenediamine (77.6 mg, 0.22 mmol), 30% hydrogen peroxide by mass concentration (14 μL, 0.85 mmol), ammonium cerium nitrate (60.6 mg, 0.11 mmol), and heat at 55 °C for 2 h. After the reaction is complete, the post-treatment method is the same as that of compound 1 to obtain compound 15 with a yield of 41.3%. 1 H-NMR (400 MHz, CDCl 3 ) δ 8.02 (d, 8.7 Hz, 2H), 7.57 - 7.59 (m, 2H), 7.18 - 7.23 (m, 3H), 6.94 (d, J = 7.6 Hz, 1H), 6.90 (s, 1H), 6.85 (d, J = 8.7 Hz, 2H), 6.78 - 6.80 (m, 1H), 6.49 (d, J = 15.8 Hz, 1H), 6.23 - 6.30 (m, 1H), 4.06 (t, J = 5.5 Hz, 2H), 3.79 (s, 3H), 3.20 (d, J = 6.7 Hz, 2H), 2.80 (t, J = 5.4 Hz, 2H), 2.68 (s, 8H). 13 C-NMR (100 MHz, CDCl 3 ) δ 160.3, 159.9, 152.1, 138.0, 134.3, 129.7, 128.4, 125.2, 122.7, 122.6, 119.2, 115.1, 113.7, 111.6, 65.9, 60.7, 56.9, 55.3, 53.1, 52.7. HRMS (ESI-TOF) m / z calcd. for C 29 H32 N 4 O 2 [M+Na] + : 491.2417, found 491.2411. The 1H NMR spectrum of Compound 15 is shown in Figure 15 .

[0161] Anti-tumor Biological Activity Evaluation of Compounds 1-15 and 1a in Example 16

[0162] (1) Tumor cells in the logarithmic growth phase (including hepatoma cells HepG2, Huh7, normal hepatocytes LO2, triple-negative breast cancer MDA-MB-231 cells, ovarian cancer A2780 cells, and cisplatin (DPP)-resistant ovarian cancer A2780 cells) were added to trypsin for digestion, centrifuged and collected. The cell suspension was diluted with medium for cell counting, and the cell density was adjusted to 5×10 4 cells / mL.

[0163] (2) The prepared cell suspension was mixed well and added to a 96-well plate, 100 μL was added to each well, and it was placed in a 5% CO 2 incubator and cultured at 37 °C for 24 h.

[0164] (3) The test compounds and the positive control camptothecin were dissolved in dimethyl sulfoxide (DMSO). Different concentrations (100, 10, 1, 0.1, 0.01, 0.001, 0 μM) of the test compounds and camptothecin were added to the above 96-well plate, 0.5 μL was added to each well, and three replicates were set for each different concentration of each drug. After adding the drugs, the 96-well plate was placed in the incubator and cultured for another 48 h.

[0165] (4) The pre-prepared thiazolyl blue (MTT) solution was added under light-proof conditions, and the sample volume added to each well was 20 μL, with a concentration of 5 mg / mL.

[0166] (5) The 96-well plate was placed in the incubator. After the MTT solution acted for 4 h, the supernatant in the wells was removed, and 100 μL of DMSO was added to each well.

[0167] (6) It was shaken at a constant temperature of 37 °C for 10 min. Using an enzyme-labeled instrument, the OD value of each well was detected at two wavelengths of 492 nm and 630 nm, and the cell survival rate was calculated as follows:

[0168] Cell survival rate (%) = (OD of experimental group - OD of blank group) / (OD of control group - OD of blank group) × 100%

[0169] The in vitro anti-tumor activity results of Compounds 1-15 and 1a are shown in Table 1.

[0170] Table 1 Activities of Compounds 1-15 and 1a in Inhibiting Proliferation of Hepatocarcinoma Cells In Vitro

[0171]

[0172]

[0173]

[0174] Note: a The experimental data are the means of three independent parallel experiments; b Camptothecin is the positive control drug; NT = not tested.

[0175] The results in Table 1 show that the conjugates of cinnamyl piperazine and 2-phenylbenzimidazole and their derivatives all have certain inhibitory activities against HepG2 cells. Among them, Compounds 1, 1a, 2, 4-6, 11, 12, and 15 have relatively high activities, with IC 50 less than 1 μM. Analyzing from the structure-activity relationship, the modification of the benzimidazole ring has a greater impact on the activity. The absence of substituents on the benzene ring of benzimidazole or the introduction of F, CF 3 , CN, and COOCH 3 is beneficial to improving the activity (comparing Compounds 2, 4-6 with 1); the connecting bridge between cinnamyl piperazine and benzimidazole should not be too long. For example, the activities of Compounds 7 and 8 are dozens of times lower than that of Compound 1; while the introduction of a methoxy group on the benzene ring of cinnamyl piperazine has no effect on the activity (comparing Compound 15 with 1). In addition, for the compounds with good activity (IC 50 less than 1 μM), the activities against another hepatocarcinoma cell line, Huh7 cells, were further tested. The results showed that these compounds also had inhibitory abilities against Huh7 cells. Although the activities were not as sensitive as those against HepG2 cells, they were all higher than the inhibitory activity of the positive control camptothecin (IC 50 > 100 μM) against Huh7 cells.

[0176] Particularly noteworthy is that this series of compounds has relatively low toxicity to normal liver LO2 cells (IC 50 3-51 μM), which are far lower than the toxicity of the positive control camptothecin (IC 50 = 0.02 μM). For example, the toxicity of Compound 1 with high activity against HepG2 cells (IC 50 = 0.18 μM) and the toxicity against normal liver LO2 cells (IC 50 = 35.38 μM) differ by about 200 times. Compounds 2-8 and 11-15 all have differences in orders of magnitude. This result indicates that this series of compounds has good selectivity and low toxicity, and to a certain extent, can make up for the deficiencies of existing chemotherapeutic drugs.

[0177] In addition, compound 1a, as the hydrochloride salt of compound 1, has similar activity to compound 1, but its water solubility is significantly improved. After testing, the water solubility of compound 1a is greater than 10 mg / mL, while that of compound 1 is less than 0.1 mg / mL. Additionally, see attached Figure 16 , the 1H NMR spectrum of compound 1a was measured after being dissolved in deuterium oxide.

[0178] The inhibitory activities of this series of compounds against triple-negative breast cancer cells MDA-MB-231 were further tested, and the inhibitory activities of three representative compounds 1, 12, and 15 against ovarian cancer cells A2780 and cisplatin-resistant A2780 cells (A2780 / DDP) were selected. The results are shown in Table 2.

[0179] Table 2. In vitro inhibitory activities of compounds 1-15 and 1a against the proliferation of triple-negative breast cancer and ovarian cancer

[0180]

[0181]

[0182] Note: a The experimental data are the means of three independent parallel experiments; b Camptothecin is the positive control drug for ovarian cancer cells; c Paclitaxel is the positive control drug for triple-negative breast cancer.

[0183] As can be seen from Table 2, compounds 1-15 also have certain inhibitory activities against triple-negative breast cancer. At high concentrations, they are even comparable to paclitaxel, and compounds 1, 4, 5, and 15 are also equivalent to paclitaxel at low concentrations. In addition, three representative compounds 1, compound 12 (with substituents on the imidazole nitrogen), and compound 15 (with substituents on the phenyl ring of cinnamyl piperazine) were selected to test their inhibitory activities against ovarian cancer cells A2780 and cisplatin-resistant A2780 cells (A2780 / DDP). The results showed that all three compounds had inhibitory activities. In particular, the activity of compound 1 against drug-resistant ovarian cancer A2780 / DDP cells was equivalent to that of the positive control drug camptothecin.

[0184] Example 17. Effect of compound 1 on the longitudinal migration ability of cisplatin-resistant ovarian cancer cells

[0185] The effect of compound 1 on the longitudinal migration ability of cisplatin-resistant ovarian cancer (A2780 / DDP) cells was evaluated using the Transwell assay. The method is as follows:

[0186] (1) The Transwell chamber was irradiated in a laminar flow hood for 30 min. First, 600 μL of medium containing 20% serum was added to the lower chamber of the chamber. Cells in the logarithmic growth phase were taken and resuspended in serum-free cell solution at a density of 1×105 Spread evenly on the upper chamber of the chamber at a density of 5 cells / mL, 200 μL for each upper chamber.

[0187] (2) After the cells adhered to the wall, add 1 μL of compound 1 at different concentrations (0, 5, 10 μM).

[0188] (3) After 24 h, first discard the culture media in both the upper and lower chambers, wash the inside and outside of the chamber with phosphate buffer (1×PBS) for 3 times in total.

[0189] (4) Fix with 4% paraformaldehyde at room temperature for 30 min, aspirate the 4% paraformaldehyde, and wash with 1×PBS twice. Add 0.1% crystal violet solution to both the upper and lower chambers and stain for 10 min.

[0190] (5) After staining, wash the chamber with 1×PBS, air-dry and then take pictures.

[0191] At the tested concentrations, compound 1 could significantly inhibit the longitudinal migration of A2780 / DDP cells. Further statistical analysis was performed on the number of migrated cells (as shown in Figure 17 ), and there were significant differences at all tested concentrations. The numbers of migrated cells of compound 1 at 5 and 10 μM concentrations were 102 and 10 respectively, showing significant differences compared with the solvent blank control DMSO group (198) (***p < 0.001), indicating that compound 1 could significantly inhibit the longitudinal migration of cisplatin-resistant ovarian cancer cells.

[0192] In summary, a series of cinnamylpiperazine and 2-phenylbenzimidazole conjugates were synthesized in the present invention, and the in vitro anti-tumor activities of these compounds were evaluated. The synthesized compounds are all new compounds with high novelty. The synthesis process of the present invention is simple, with high reaction yield and short time consumption. The activity study shows that these compounds can inhibit the proliferation of various tumor cells, and some compounds have equivalent activities to the positive control. In particular, it is worth mentioning that these compounds have low toxicity and high selectivity for tumor cells. Cisplatin-resistant ovarian cancer has always been a difficult problem in clinical treatment, with a high recurrence rate and low survival rate of patients. The compounds in the present invention make up for the shortage of such drugs. In addition, these compounds can be made into hydrochloride salts, with significantly improved water solubility and good drug-forming properties. Therefore, it shows that these compounds have good application prospects.

[0193] Although the embodiments of the present invention are disclosed for illustrative purposes, those skilled in the art can understand that various substitutions, changes and modifications are possible without departing from the spirit and scope of the present invention and the appended claims. Therefore, the scope of the present invention is not limited to the content disclosed in the embodiments.

Claims

1. An antitumor cinnamyl piperazine-2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof, characterized in that: Its general structure is as follows: Wherein, X on the connecting bridge = oxygen, nitrogen, sulfur, carbon, n = 1 to 6; R1 is hydrogen, methoxy, hydroxyl, trifluoromethyl, fluorine, chlorine, cyano, formic acid, methyl formate, ethyl formate, methyl, ethyl, propyl, butyl, carbamoyl, methylaminocarbamoyl, cyclopropylaminocarbamoyl, propylaminocarbamoyl, R2 is hydrogen, acetyl, methyl, ethyl, propyl, butyl, butyronitrile, propionitrile, acetonitrile; R3 is hydrogen, methoxy, hydroxyl, methyl, ethyl, propyl, butylmethylamino, dimethylamino substituted at positions 1' to 3'.

2. The antitumor cinnamyl piperazine-2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The structural formula of the anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof is one of the following:

3. The general method for preparing the antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that: The steps include: First, N-tert-butyloxycarbonyl (Boc) piperazine is connected with cinnamyl bromide to obtain compound 15, and the Boc protecting group is removed to obtain compound 16; triethylene glycol and its analogs are brominated to obtain compound 17; p-hydroxybenzaldehyde is reacted with 1,2-dibromoethane, 2,2'-dibromodiethyl ether, compound 17 and its analogs to obtain corresponding ethers 18-20; compound 16 is further connected to obtain compounds 21-23 with different connecting bridge lengths; finally, it is reacted with substituted o-phenylenediamine to obtain final products 1-8, and hydrogen chloride gas is introduced into methanol or acetone solution to obtain the corresponding hydrochloride.

4. The general preparation method according to claim 3, characterized in that: The reaction route is: Alternatively, 3,4-diaminobenzoic acid is reacted with cyclopropylamine to obtain 3,4-diamino-N-cyclopropylbenzamide (24), which is reacted with compound 21 to obtain the final product 9. The reaction route is: Alternatively, the cyano group of compound 6 is hydrolyzed to obtain compound 10, and the reaction route is: Compound 1 reacts with an alkyl halide under alkaline conditions to obtain imidazole nitrogen derivative products 11 to 13; or, compound 1 reacts with an acyl chloride to obtain amide 14, and the reaction route is: Alternatively, a substituted cinnamyl piperazine intermediate 30 is prepared through a series of reactions, and then reacted with compound 19 through similar steps to obtain compound 31, and finally reacted with o-phenylenediamine to obtain compound 15. The reaction route is:

5. Use of the antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 in the preparation of a drug for treating liver cancer.

6. Use of the antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 in the preparation of a drug for treating breast cancer and triple-negative breast cancer.

7. Use of the antitumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 in the preparation of a drug for treating ovarian cancer and drug-resistant ovarian cancer.

8. Use of the anti-tumor cinnamyl piperazine and 2-phenylbenzimidazole conjugate or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 in the preparation of a drug for treating tumors.

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