Benzimidazolyl phenyl thiazole copper complex as well as synthesis method and application thereof

Abstract

The invention belongs to the technical field of antitumor drug research, and relates to a benzimidazolyl phenyl thiazole copper complex as well as a synthesis method and application of the benzimidazolyl phenyl thiazole copper complex. The benzimidazolyl phenyl thiazole copper complex CuL2Cl is synthesized from 2, 6-dimethyl-1H-benzimidazole-2-yl)-4-phenyl thiazole and copper chloride for the first time, and the benzimidazolyl phenyl thiazole copper complex CuL2Cl is synthesized from 2, 6-dimethyl-1H-benzimidazole-2-yl)-4-phenyl thiazole. Wherein L is 2-(5, 6-dimethyl-1H-benzimidazole-2-yl)-4-phenyl thiazole, and the structural formula is shown in the description. A test result shows that the copper complex has an inhibiting effect on a human liver cancer cell line HuH7, has the capability of inhibiting cancer cell invasion and migration, has the capability of inducing apoptosis of cancer cells and the capability of inducing cycle arrest of cancer cells, and can be used for preparing a medicine for treating liver cancer. The benzimidazolyl phenyl thiazole copper complex is proved to be an ideal novel anti-cancer drug.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of anti-tumor drugs, and particularly relates to a benzimidazolyl phenyl thiazole copper complex as well as a synthesis method and application thereof. BACKGROUND

[0002] Since cisplatin was first developed as a metal-based anti-tumor drug, the research and development of metal-based chemotherapy drugs has opened up an innovative direction. Although platinum drugs have shown significant efficacy in clinical applications, their inherent drug resistance mechanisms and systemic toxicity problems have not been fundamentally solved. Under this background, the development of new metal complexes that break through traditional platinum drugs has become an important research direction in the field of medicinal chemistry. The research in this field focuses on the construction of metal center complexes with multiple advantages, including optimizing drug selectivity to reduce toxic side reactions, breaking through the tumor drug resistance barrier, and expanding the anti-tumor spectrum, and other key scientific issues.

[0003] In terms of the development of new metal center ligands, benzimidazole and its derivatives have attracted attention due to their unique molecular structure. This class of imine compounds, which are generated by the condensation of amino and carbonyl groups, has become an important building block for metal drug research and development due to its flexible coordination ability and controllable electronic properties. Among them, benzimidazole compounds, as functional nitrogen heterocyclic derivatives, have intramolecular N donor atoms that can form stable chelate systems through coordination bonds, and also have special biological activity. This unique structure and activity makes it show significant advantages in the treatment of anti-tumor, antibacterial, antiviral and other fields. It is worth noting that preclinical studies have confirmed that some benzimidazole compounds have anti-tumor activity, providing a structural basis for the development of new metal center complexes. However, the metabolic pathway of this class of drugs has the risk of off-target toxicity, and cannot maintain controllable release within the physiological concentration range, which may cause metal ion accumulation and cause liver and kidney damage, thereby shortening the treatment window.

[0004] Based on this, the research team designed and synthesized a new type of benzimidazolyl phenyl thiazole copper complex. The new benzimidazolyl phenyl thiazole copper complex significantly improves the anti-tumor efficacy while maintaining good safety. SUMMARY

[0005] In view of the above technical problems, the purpose of the present application is to provide a benzimidazolyl phenyl thiazole copper complex and a synthesis method and application thereof. The compound integrates the DNA intercalation ability of benzimidazole and the electron transfer properties of thiazole ring, and constructs an anti-tumor system with multiple mechanisms, which can significantly improve the anti-tumor efficacy while maintaining good safety.

[0006] To solve the above technical problems, the present application provides a benzimidazolyl phenyl thiazole copper complex, and the chemical molecular formula of the benzimidazolyl phenyl thiazole copper complex is: 36H 32 Cl2CuN6S2, having the chemical formula: CuL2Cl, wherein the structure of the CuL2Cl is: wherein L in the chemical formula is 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole.

[0007] Preferably, the crystal space group of the benzimidazolyl phenylthiazole copper complex is P21 / c, the unit cell volume is 3825.14(12), Z=4, Mr=818.33 Da, Dx=1.421 g / cm 3 .

[0008] The application provides a preparation method of a benzimidazolyl phenylthiazole copper complex, characterized by comprising the following steps: The ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole is mixed with copper chloride dihydrate and heated to dissolve in a methanol solvent to perform a coordination reaction, and yellow block crystals, i.e., the benzimidazolyl phenylthiazole copper complex, are obtained.

[0009] Preferably, after the coordination of the ligand and the copper metal center, a mode molecule with one metal center and two ligand molecules is generated, and the molar ratio of 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to copper chloride dihydrate is 1:1-2.

[0010] Preferably, the coordination reaction can smoothly occur under room temperature and heating conditions, the temperature is 25-70 DEG C, and the heating condition can realize rapid reaction to generate the target metal complex, and the solvent evaporation process under room temperature is slow, so a long reaction time is needed, and therefore the length of the coordination reaction is 24-72 hours.

[0011] Preferably, the molar volume ratio of the ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to methanol is 1:6-1:10.

[0012] The application provides an application of the benzimidazolyl phenylthiazole copper complex in preparation of an anti-hepatoma tumor drug.

[0013] Preferably, the anti-hepatoma tumor drug has a therapeutic effect on human hepatoma cells HuH7.

[0014] An anti-tumor drug, the drug comprising the benzimidazolyl phenylthiazole copper complex in claim 1 and a pharmaceutical excipient.

[0015] Compared with the prior art, the application has the following beneficial effects: The benzimidazolyl phenyl thiazole copper complex CuL2Cl given by the present application is synthesized by first coordinating the benzimidazolyl phenyl thiazole ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole with copper chloride. Compared with platinum drugs, the copper in the copper complex is a trace element necessary for the human body, and its metabolic pathway can reduce the risk of off-target toxicity, because the copper complex generally maintains controlled release in the physiological concentration range through chelation, avoiding liver and kidney damage caused by metal ion accumulation, thereby widening the therapeutic window. In addition, the copper complex can not only directly inhibit tumor proliferation through DNA damage, but also target key enzyme systems or regulate tumor microenvironments. Therefore, the copper complex has an inhibitory effect on human hepatoma cell line HuH7, and has the ability to inhibit cancer cell invasion and migration, the ability to induce cancer cell apoptosis, and the ability to induce cancer cell cycle arrest, proving that the benzimidazolyl phenyl thiazole copper complex is an ideal new type of anticancer drug.

[0016] The synthesis process of the benzimidazolyl phenyl thiazole copper complex provided by the present application is simple, low in cost, easy to control chemical components, good in repeatability and high in yield. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 It is a minimum asymmetric unit crystal structure diagram of the copper complex of the present application.

[0018] Figure 2 It is a flow cytometry result diagram of the Rh123 staining of the copper complex on human hepatoma cells HuH7.

[0019] Figure 3 It is a flow cytometry result diagram of the change of ROS in human hepatoma cells HuH7 treated by the copper complex.

[0020] Figure 4 It is a statistical analysis diagram of the scratch test result of the copper complex on human hepatoma cells HuH7.

[0021] Figure 5 It is a flow cytometry result diagram of the apoptosis of human hepatoma cells HuH7 treated by the copper complex.

[0022] Figure 6 It is a result diagram of the invasion of human hepatoma cells HuH7 treated by the copper complex.

[0023] Figure 7 It is a result diagram of the change of the weight of mouse xenograft tumor of human hepatoma cells HuH7 treated by the copper complex.

[0024] Figure 8 It is a result diagram of the change of the volume of mouse xenograft tumor of human hepatoma cells HuH7 treated by the copper complex. DETAILED DESCRIPTION

[0025] The specific embodiments of the present application are described in detail below, but the scope of protection of the present application is not limited by the specific embodiments. Based on the examples in the present application, all other examples obtained by those of ordinary skill in the art without creative labor fall within the scope of protection of the present application. The experimental methods described in the embodiments of the present application are conventional methods unless otherwise specified.

[0026] It should be noted that when numerical ranges are involved in the present application, both endpoints of each numerical range and any number between the two endpoints can be selected. Since the steps and examples 1-6 are the same, in order to avoid repetition, the present application describes preferred examples, but the present application is not limited thereto, but can also be embodied in other ways within the scope of the technical solutions defined in the appended claims. The various raw materials, reagents, instruments and equipment used in the following embodiments of the present application can be purchased from the market or prepared by existing methods.

[0027] The technical solutions in the embodiments of the present application are described clearly and completely in combination with the preferred embodiments and the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present application, not all. Based on the examples in the present application, all other examples obtained by those of ordinary skill in the art without creative labor fall within the scope of protection of the present application.

[0028] Among the non-platinum metal candidates, copper-based complexes exhibit unique development value. As an endogenous essential trace element in the human body, copper ions serve as the core catalytic component of various metalloenzymes, regulate electron transport chain reactions and redox balance, and play an irreplaceable biological function in cell cycle regulation and immune homeostasis maintenance. By binding to the active centers of key enzymes such as superoxide dismutase (SOD) and cytochrome c oxidase, copper ions are directly involved in important physiological processes such as mitochondrial energy metabolism, DNA synthesis and repair, and immune cell differentiation. This unique redox regulation feature makes copper ions a core mediator connecting basic metabolism and immune response, and its imbalance has been proven to be significantly associated with tumor occurrence and development. From the perspective of medicinal chemistry, the advantages of copper complexes are reflected in the following aspects: first, copper ions (Cu + / Cu 2+) unique redox properties enable it to generate reactive oxygen species (ROS) through Fenton-like reactions, selectively inducing tumor cell apoptosis. Compared with normal cells, oxidative stress imbalance is prevalent in tumor cells, with a lower tolerance threshold for ROS, so copper complexes can achieve precise killing through this mechanism while reducing damage to healthy tissues; secondly, copper complexes can adapt to various coordination configurations (Jahn-Teller effect) such as tetrahedron, planar quadrilateral, and triangular bipyramid. By regulating the structure of the ligand (such as introducing N and S-containing aromatic heterocycles), the lipophilic-hydrophilic partition coefficient, membrane penetration ability, and target affinity of the complex can be optimized, and the stability of the complex can be improved by using polydentate ligands; thirdly, compared with platinum drugs, copper, as an essential trace element in the human body, can reduce the risk of off-target toxicity. Copper complexes usually maintain controlled release within the physiological concentration range through chelation, avoiding liver and kidney damage caused by metal ion accumulation, thereby widening the therapeutic window; fourthly, copper complexes not only directly inhibit tumor proliferation through DNA damage but also target key enzyme systems or regulate the tumor microenvironment; fifthly, some copper complexes have photosensitive properties and can trigger photodynamic effects under specific wavelength light or continuously generate reactive oxygen species through chemical kinetics therapy, achieving multimodal synergistic therapy; sixthly, the radioactive properties of copper isotopes (such as 64 Cu) can be used for drug metabolism tracing and integrated diagnosis and treatment design. Currently, copper complexes (such as Casiopeinas® series) have entered the clinical trial stage, showing controllable toxicokinetic characteristics and antitumor activity, which provides a theoretical basis for the development of copper-based antitumor drugs.

[0029] Compared with other transition metals, copper complexes have a significant advantage in biocompatibility. Copper-based drugs not only have lower cytotoxicity, but their unique redox regulation mechanism can also selectively induce tumor cell apoptosis. Based on this, our research team designed and synthesized a new type of benzimidazole-based phenylthiazole copper complex. This compound integrates the DNA intercalation ability of benzimidazole and the electron transfer properties of the thiazole ring, creating an antitumor system with multiple mechanisms of action, while maintaining good safety and significantly improving antitumor efficacy. Among them, the present application provides a benzimidazole-based phenylthiazole copper complex, which has the chemical molecular formula of: 36 H 32 Cl2CuN6S2, with the following chemical formula: CuL2Cl, and the structural formula of CuL2Cl is:

[0030] wherein L in the chemical formula is 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole.

[0031] The crystal space group of the benzimidazolylphenylthiazole copper complex is P21 / c, with a unit cell volume of 3825.14(12), Z = 4, Mr = 818.33 Da, and Dx = 1.421 g / cm³. 3 .

[0032] To enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments: Example 1 The preparation steps of a benzimidazole-phenylthiazole copper complex are as follows: The ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole was mixed with copper chloride CuCl2∙2H2O at a molar ratio of 1:1 and dissolved in 6 mL of methanol solution (the molar volume ratio of 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to methanol was 1:6). The entire mixture was placed in a glass bottle and reacted at 25 °C for 72 h to obtain yellow blocky crystals. The crystals were filtered and dried at room temperature. Yield: 83%.

[0033] Example 2 The preparation steps of a benzimidazole-phenylthiazole copper complex are as follows: The ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole was mixed with copper chloride CuCl2∙2H2O at a molar ratio of 1:1 and dissolved in 8 mL of methanol solution (the molar volume ratio of 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to methanol was 1:8). The entire mixture was placed in a glass bottle and reacted at 70 °C for 48 h. The reaction solution was slowly cooled to room temperature to obtain yellow blocky crystals, which were filtered and dried at room temperature. Yield: 77%.

[0034] Example 3 The preparation steps of a benzimidazole-phenylthiazole copper complex are as follows: The ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole was mixed with copper chloride CuCl2∙2H2O at a molar ratio of 1:1 and dissolved in 10 mL of methanol solution (the molar volume ratio of 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to methanol was 1:10). The entire mixture was placed in a glass bottle and reacted at 25 °C for 72 h to obtain yellow blocky crystals. The crystals were filtered and dried at room temperature. Yield: 85%.

[0035] Example 4 The preparation steps of a benzimidazole-phenylthiazole copper complex are as follows: The ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole was mixed with copper chloride CuCl2∙2H2O at a molar ratio of 1:2 and dissolved in 6 mL of methanol solution (the molar volume ratio of 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to methanol was 1:6). The entire mixture was placed in a glass bottle and reacted at 70 °C for 48 h. The reaction solution was slowly cooled to room temperature to obtain yellow blocky crystals, which were filtered and dried at room temperature. Yield: 81%.

[0036] Example 5 A benzimidazole-phenylthiazole copper complex was prepared according to the following steps: The ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole was mixed with copper chloride CuCl2∙2H2O at a molar ratio of 1:2 and dissolved in 8 mL of methanol solution (the molar volume ratio of 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to methanol was 1:8). The entire mixture was placed in a glass bottle and reacted at 50 °C for 24 h to obtain yellow blocky crystals. The crystals were filtered and dried at room temperature. Yield: 83%.

[0037] Example 6 The preparation steps of a benzimidazole-phenylthiazole copper complex are as follows: The ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole was mixed with copper chloride CuCl2∙2H2O at a molar ratio of 1:2 and dissolved in 10 mL of methanol solution (the molar volume ratio of 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to methanol was 1:10). The entire mixture was placed in a glass bottle and reacted at 50 °C for 24 h. The reaction solution was slowly cooled to room temperature to obtain yellow blocky crystals, which were filtered and dried at room temperature. Yield: 79%.

[0038] The benzimidazole-phenylthiazole copper complexes were prepared in all of the above Examples 1 to 6. Example 1 is selected as an example to illustrate the performance and effects of the benzimidazole-phenylthiazole copper complexes provided by the present invention.

[0039] Experimental verification (1) Performance verification of benzimidazole phenylthiazole copper complex The yellow blocky crystals (benzimidazolylphenylthiazole copper complex) prepared in Example 1 were analyzed using X-ray single-crystal diffraction. The X-ray single-crystal diffraction data indicated that the copper complex belongs to the P21 / c space group. Figure 1As shown, its smallest asymmetric unit is a small molecule composed of a copper atom, two ligands, and a coordinated chlorine atom. The copper atom follows a five-coordinate irregular tetragonal pyramidal coordination model. The nitrogen and chlorine atoms of ligand L are coordinated with the copper atom. The two ligands L exhibit a certain twist angle.

[0040] Single crystals with good crystalline forms were selected and subjected to single-crystal testing using an X-ray single-crystal diffractometer. The crystal structure was then analyzed using Olex2 software. The results showed that the copper complex had a space group of P21 / c, a cell volume of 3825.14(12), Z = 4, Mr = 818.33 Da, and Dx = 1.421 g / cm³. 3 .

[0041] (2) Verification of the efficacy of benzimidazole-phenylthiazole copper complex ① The specific procedure for staining human hepatocellular carcinoma cells HuH7 with benzimidazole-phenylthiazole copper complex is as follows: Cells were loaded at 2×10 5 Cells were seeded at a density of 1 / 2 well into 6-well cell culture plates. After the cells adhered and grew to a suitable density, the old culture medium was removed, and fresh medium containing a copper complex solution was added at a volume of 2.5 mL / well. The concentrations of the same complex used were 0 μM and 32 μM. After treating the cells for 48 h, Rh123 reagent was added to the culture plate at a volume of 0.5 mL / well. The cells were incubated at 37°C for 20 min, and then collected and analyzed by flow cytometry. Experimental results are as follows: Figure 2 As shown.

[0042] Through such Figure 2 The Rh123 staining results of the benzimidazole-phenylthiazole copper complex on human hepatocellular carcinoma cells (HuH7) show that low-intensity peaks were observed in HuH7 cells under the influence of the benzimidazole-phenylthiazole copper complex, indicating that mitochondrial function was disrupted. The results demonstrate that the benzimidazole-phenylthiazole copper complex can effectively disrupt mitochondrial homeostasis in HuH7 tumor cells.

[0043] ② The effects of the benzimidazole-phenylthiazole copper complex on ROS changes in human hepatocellular carcinoma cells HuH7 are described in the following procedure: Cells were loaded at 2×10 5 Cells were seeded at a density of 1 / 2 well in 6-well cell culture plates. After the cells adhered and grew to a suitable density, the old culture medium was removed, and fresh medium containing copper complex solution was added at a volume of 2.5 mL / well. The concentrations used were 0 μM and 15 μM. After 48 h, DCFH-DA reagent was added to the culture plate at a volume of 0.5 mL / well, and the plates were incubated at 37 °C for 20 min. Cells were then collected and analyzed by flow cytometry. The experimental results are as follows: Figure 3 As shown.

[0044] Through such Figure 3 The DCFH-DA staining results of the benzimidazolylphenylthiazole copper complex on human hepatocellular carcinoma cells HuH7 show that, in the cellular ROS quantification analysis, a red shift in the green fluorescence intensity of HuH7 cells was observed under the influence of the copper complex, indicating an increase in intracellular ROS levels. The results demonstrate that the copper complex can increase intracellular ROS levels and enhance intracellular oxidative damage in HuH7 tumor cells.

[0045] ③The steps for obtaining the scratch assay results of the benzimidazole-phenylthiazole copper complex against human hepatocellular carcinoma cells HuH7 are as follows: The effect of benzimidazole-phenylthiazole copper complex on the migration ability of human hepatocellular carcinoma cell line HuH7 was investigated by seeding HuH7 cells into 6-well plates. After the cells reached 80% confluence under a microscope, parallel straight lines were rapidly drawn in the middle of each well using a 10 μL pipette tip. The culture medium was discarded, and the cells were washed three times with PBS. Cells were then treated with 0 μM and 3 μM benzimidazole-phenylthiazole copper complex for 48 h. The scratched areas were observed and photographed under a microscope at 0 h, 24 h, and 48 h after treatment. The experimental results are as follows: Figure 4 As shown.

[0046] Through such Figure 4 The results of the scratch assay on human hepatocellular carcinoma cells HuH7, presented by the benzimidazole-phenylthiazole copper complex, show that the healing area of ​​HuH7 cells was significantly reduced under the action of the copper complex, indicating that cell migration ability was also inhibited. The results demonstrate that the copper complex can effectively block the healing of HuH7 tumor cells and has a good inhibitory effect on the in vitro migration of human hepatocellular carcinoma cells.

[0047] ④ The experimental steps for inducing apoptosis in human hepatocellular carcinoma cells HuH7 by the copper complex of benzimidazole phenylthiazole are as follows: Tumorigenesis is closely related to the dysregulation of apoptosis. To investigate whether benzimidazole-phenylthiazole copper complex effectively kills HuH7 tumor cells by inducing apoptosis, HuH7 cells were treated with 0 μM and 16 μM concentrations of benzimidazole-phenylthiazole copper complex for 48 h, then washed twice with pre-cooled PBS buffer, and 5 μL of culture medium containing Annexin V-FITC fluorescent probe was added. The cells were incubated in the dark for 20 min. Then, 5 μL of PI fluorescent probe was added, and the cells were cultured for another 20 min. Finally, flow cytometry was used for analysis. The experimental results are as follows: Figure 5 As shown.

[0048] pass Figure 5It was found that, compared with the control group, 60.6% of HuH7 cells treated with the benzimidazolylphenylthiazole copper complex underwent apoptosis. The experimental results are shown in […]. Figure 5 The experimental results show that the benzimidazolylphenylthiazolium copper complex has significant apoptosis-inducing activity against human hepatocellular carcinoma cells HuH7.

[0049] ⑥ The experimental steps for the effect of copper complexes on the invasion of human liver cancer cells HuH7 are as follows: To investigate the inhibitory effect of drugs on cell invasion, Transwell chambers were first placed in 24-well plates. The upper chamber was pre-incubated with serum-free medium for 1 hour to equilibrate the environment. For the invasion assay, Matrigel was pre-coated at the bottom of the chamber. Subsequently, logarithmic growth phase HuH7 cells were digested and resuspended in serum-free medium, adjusting the density to 1×10⁶ cells / well. 6 After reaching a cell / mL concentration, 200 μL of the suspension was added to the upper chamber, and medium containing 10% FBS was added to the lower chamber. The system was incubated at 37°C in a 5% CO2 incubator to allow cell adhesion. Then, the cell culture medium in the lower chamber was replaced with cell culture medium containing 0 μM and 3 μM benzimidazolylphenylthiazolium copper complexes. After culturing the cells for 48 h, the medium was removed, and unmigrated cells in the upper chamber were wiped with a cotton swab. The cells were fixed with methanol for 30 minutes, stained with 0.1% crystal violet for 20 minutes, and washed with PBS. Finally, images were taken under a microscope. The experimental results are as follows: Figure 6 As shown.

[0050] pass Figure 6 It can be seen that after treating human liver cancer cells HuH7 cells with 0 μM and 3 μM concentrations of benzimidazole phenylthiazole copper complex for 48 h, the treatment containing benzimidazole phenylthiazole copper complex inhibited the cell invasion ability. Therefore, benzimidazole phenylthiazole copper complex can block the invasion process of human liver cancer cells HuH7 cells.

[0051] ⑦ The experimental procedure for the effect of benzimidazole-phenylthiazole copper complex on the weight change of human liver cancer cell line HuH7 xenograft tumors in mice is as follows: A mouse xenograft model of human hepatocellular carcinoma cell line HuH7 was established using 5-week-old immunodeficient mice housed in an SPF environment. After resuscitation, HuH7 cells were expanded to the logarithmic growth phase in DMEM medium containing 10% fetal bovine serum. Cells were then collected by trypsin digestion and the concentration was adjusted to 1×10⁻⁶. 7 Cells / mL: 100 μL of cell suspension was mixed thoroughly with an equal volume of matrix gel, and 200 μL of the cell suspension was injected subcutaneously into the right axilla of mice using an insulin syringe. Five mice were injected per group, with a negative control included. Mice were observed daily after inoculation; tumor formation was observed after 10 days, and the mice's weight and health status were monitored. When the tumor volume reached 50 mm², the tumor was considered closed. 3The mice were treated with medication every two days for nine consecutive days. After treatment, the mice were euthanized by spinal dislocation, and the tumor tissue was separated and weighed. The tumor weight results are as follows: Figure 7 As shown.

[0052] pass Figure 7 It was found that after 9 days of treatment with benzimidazole phenylthiazole copper complex, the average tumor weight of mouse xenografts of human liver cancer cells HuH7 was significantly lower than that of the negative control group, indicating that benzimidazole phenylthiazole copper complex can inhibit the growth of human liver cancer cells HuH7.

[0053] ⑧ The experimental procedure for the effect of benzimidazole-phenylthiazole copper complex on the volume of mouse xenograft tumors of human liver cancer cells HuH7 is as follows: A mouse xenograft model of human hepatocellular carcinoma cell line HuH7 was established using 5-week-old immunodeficient mice housed in an SPF environment. After resuscitation, HuH7 cells were expanded to the logarithmic growth phase in DMEM medium containing 10% fetal bovine serum. Cells were then collected by trypsin digestion and the concentration was adjusted to 1×10⁻⁶. 7 Cells / mL: 100 μL of cell suspension was mixed thoroughly with an equal volume of matrix gel, and 200 μL of the cell suspension was injected subcutaneously into the right axilla of mice using an insulin syringe. Five mice were injected per group, with a negative control included. Mice were observed daily after inoculation; tumor formation was observed after 10 days, and the mice's weight and health status were monitored. When the tumor volume reached 50 mm², the tumor was considered closed. 3 Drug treatment was administered every two days for nine consecutive days. During the drug treatment period, the tumor volume of a mouse xenograft model of human hepatocellular carcinoma cell line HuH7 was measured, and the results are shown below. Figure 8 As shown.

[0054] pass Figure 8 It can be seen that in mice with human liver cancer cells HuH7 xenograft tumors, the average tumor volume of the tumors in the treatment group was significantly inhibited after 9 days of treatment with benzimidazole phenylthiazole copper complex compared with the negative control group. This indicates that benzimidazole phenylthiazole copper complex can significantly inhibit the growth of human liver cancer cells HuH7 tumors, and the effect is comparable to that of cisplatin control group.

[0055] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A benzimidazolylphenylthiazolium copper complex, characterized in that, The chemical formula of the benzimidazole-phenylthiazole copper complex is: C 36 H 32 Cl₂CuN₆S₂ has the following chemical formula: CuL₂Cl, and the structural formula of CuL₂Cl is: In the chemical formula, L stands for 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole.

2. The benzimidazolylphenylthiazole copper complex according to claim 1, characterized in that, The crystal space group of the benzimidazolylphenylthiazole copper complex is P21 / c, the cell volume is 3825.14(12), Z=4, Mr=818.33Da, Dx=1.421g / cm³ 3 .

3. The method for preparing the benzimidazole-phenylthiazole copper complex according to any one of claims 1 to 2, characterized in that, Includes the following steps: The ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole was mixed with copper chloride dihydrate and heated to dissolve in methanol solvent to carry out a coordination reaction, yielding yellow blocky crystals, namely the benzimidazolylphenylthiazole copper complex.

4. The method for preparing the benzimidazole-phenylthiazole copper complex according to claim 3, characterized in that, The molar ratio of the ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to copper chloride dihydrate is 1:1 to 2.

5. The method for preparing the benzimidazole-phenylthiazole copper complex according to claim 3, characterized in that, The coordination reaction is carried out at a temperature of 25℃ to 70℃ and for a duration of 24h to 72h.

6. The method for preparing the benzimidazole-phenylthiazole copper complex according to claim 3, characterized in that, The molar volume ratio of the ligand 2-(5,6-dimethyl-1H-benzimidazol-2-yl)-4-phenylthiazole to methanol is 1:6 to 1:

10.

7. The use of the benzimidazolylphenylthiazole copper complex according to any one of claims 1 to 2 in the preparation of anti-hepatocellular carcinoma drugs.

8. The application according to claim 7, characterized in that, The aforementioned anti-liver cancer drug has a therapeutic effect on human liver cancer cells HuH7.

9. An antitumor drug, characterized in that, The drug comprises the benzimidazole phenylthiazole copper complex of claim 1 and pharmaceutical excipients.

Images