Preparation method of nano copper bismuthate with pH response sonodynamic effect

Abstract

The invention discloses a preparation method of nano copper bismuthate with a pH response sonodynamic effect, the nano copper bismuthate is a novel nano particle material based on pharmaceutical chemistry, and the nano copper bismuthate is used for tumor treatment by utilizing specific excellent properties of the nano copper bismuthate. The specific preparation method of the nano-particle comprises the following steps: 1) synthesizing nano copper bismuthate CBO through a high temperature reaction, and 2) modifying carboxyl on the surface of the nano copper bismuthate CBO through a reaction with polyethylene glycol (PEG), and the finally obtained nano-particle complex CBOP has good biocompatibility, biological safety and excellent tumor treatment effect.

Description

Technical Field

[0001] The invention relates to the technical field of chemical drugs, and in particular to a preparation method and application of nano copper bismuthate with pH-responsive sonodynamic effect. Background Art

[0002] Tumors pose a serious threat to human health. Their occurrence, development and metastasis are profoundly affected by physiological redox balance. Intelligent induction of redox imbalance has become a new field for inhibiting tumor growth and spread. Its core is to maintain the redox balance of tumors by carefully coordinating the reactive oxygen species (ROS) system and the antioxidant defense (AOD) system. There are many different methods for treating tumors today, and the appropriate means are mainly selected based on the nature and stage of the tumor and the patient's physical condition. Common tumor treatment methods include surgical treatment, drug therapy, local treatment, etc. Surgical treatment is mainly aimed at benign tumors and early malignant tumors, and the treatment goal is achieved by removing tumor tissue. Drug therapy includes chemotherapy drugs, targeted drugs and immune drugs, etc., which aim to inhibit the growth and spread of tumor cells through drug action. Local treatment is used to control the development of tumors and alleviate clinical symptoms, such as radiotherapy, interventional therapy and ablation therapy. Among these methods, sonodynamic therapy, as a new type of tumor treatment method, has received widespread attention in recent years. Sonodynamic therapy uses the strong penetration ability of ultrasound into biological tissues, and focuses the sound energy non-invasively into deep tissues through focused ultrasound, activating sonosensitizers and producing anti-tumor effects. This treatment method has the advantages of being non-invasive, repeatable, and low toxic. It can promote the response of the body's immune system, thereby enhancing anti-tumor ability. The key to sonodynamic therapy lies in the selection and application of sonosensitizers. Sonosensitizers can produce lethal reactive oxygen species (ROS) and local damage under the action of ultrasound, thereby killing tumor cells. However, traditional sonosensitizers have many shortcomings, which limit the clinical application of sonodynamic therapy. Therefore, the research and development of new responsive nanosonosensitizers has become the key to achieving efficient, specific and safe treatment. In ultrasound (US)-triggered sonosensitization therapy (SDT), inorganic sonosensitizers play a key role. They can produce reactive oxygen species (ROS) through sonodynamic action, thereby reshaping the redox balance in tumors.

[0003] To date, many inorganic nanosonosensitizers have been developed and applied in anti-tumor research, such as ZnO, CuS and TiO 2etc. Copper bismuthate is a newly discovered new multifunctional tumor microenvironment (TME) regulator used in the biomedical field to regulate tumor acidity and hypoxia. On the one hand, it uses its narrow bandgap structure to generate ROS through sonosensitivity to cause oxidative damage to tumor cells for sonodynamic therapy (SDT), and uses catalytic activity to generate oxygen to relieve tumor hypoxia and enhance anti-tumor effects. At the same time, peroxidase activity (POD) can also be used to react with abnormal factors in TME, promoting the multivalent redox reaction of copper during the reaction process, converting endogenous H 2 O 2 The conversion of CuBi into cytotoxic hydroxyl radicals (·OH) causes oxidative damage for chemodynamic therapy (CDT), ultimately leading to tumor cell death. This enhanced sonodynamic therapy (SDT) and chemodynamic therapy (CDT) combined action can produce a large amount of ROS for oxidative damage. In addition, the experiment surprisingly found that CuBi 2 O 4 It also exhibits glutathione peroxidase (GPx) activity, which can consume intracellular glutathione (GSH), thereby further exacerbating redox imbalance. In summary, copper bismuthate nanomaterials, as a heavy metal compound with photosensitivity and catalytic activity, can fully utilize the production of hydrogen peroxide in the tumor microenvironment and enhance the generation of ROS through the rational design of a stable nanoplatform and synergistic with other treatment methods, such as sonosensitization therapy and chemodynamic therapy, thereby amplifying the killing effect of oxidative damage on tumor cells and inhibiting tumor growth and spread to a certain extent, and have potential anti-tumor activity. With the in-depth study of copper bismuthate nanomaterials in tumor treatment, copper bismuthate has one of the basic elements of human health and has a weak effect on the body's metabolism, making it a potential application prospect in tumor treatment. Summary of the invention

[0004] In view of the problems that the drugs currently used for sonodynamic therapy of tumors have a single effect, high preparation cost, poor treatment effect, poor biocompatibility, etc., the present invention aims to synthesize a new tumor treatment drug that is simple to prepare, has high biocompatibility, and can achieve synergistic treatment of sonodynamic therapy and chemodynamic therapy.

[0005] The technical solution of the present invention is as follows: The preparation method of nano copper bismuthate with pH-responsive sonodynamic effect is characterized by comprising the following steps: (1) Preparation method of nano-copper bismuthate with pH-responsive sonodynamic effect: First, sodium methoxide and polyvinyl pyrrolidone are added to water in a certain mass ratio to dissolve, wherein the mass ratio of sodium methoxide to polyvinyl pyrrolidone ranges from 1:1 to 1:3; then CuCl is added 2 ·H 2O was ultrasonically treated until a blue clear solution was obtained. Sodium methoxide and polyvinyl pyrrolidone were added to water in a certain mass ratio to dissolve, wherein the mass ratio of sodium methoxide to polyvinyl pyrrolidone was 1:1 to 1:2; then Bi(NO 3 ) 3 ·5H 2 O to form an emulsion. Finally, the two liquids were mixed evenly, ultrasonically dispersed for 5 minutes, stirred at room temperature for 4 hours, and then the solution was hydrothermally treated in an oven at 140°C for 6 hours. After cooling to room temperature, centrifuged at 10,000 rpm for 2 minutes, washed with deionized water 5 times, and dried to obtain copper bismuthate nanoparticles CBO; (2) Carboxyl modification of copper bismuthate nanoparticles: Using the EDC / NHS activation method, NHS / EDC / CBO was reacted at a certain molar ratio at room temperature for 12 hours, and then PEG with a mass ratio of 6:1 was added and stirred at room temperature for 12 hours. The excess material was removed by centrifugation to obtain nano-copper bismuthate with pH-responsive sonodynamic effect.

[0006] Furthermore, in step (1), CuCl 2 ·H 2 The concentration of O solution is 0.6 mmol / L.

[0007] Furthermore, in step (1), Bi(NO 3 ) 3 ·5H 2 The concentration of O was 1.2 mmol / L.

[0008] Furthermore, in the step (2), the molar ratio of CBO / EDC / NHS is 2:3:5 to 2:3:8.

[0009] The main advantages of the present invention are: In response to the problem of poor therapeutic effects of current tumor drugs, the present invention creatively prepares a material with good biocompatibility, strong stability, chemical dynamics and sonodynamic functions. In addition to the advantages of good biocompatibility and biosafety, the nano-copper bismuthate with pH-responsive sonodynamic effect in this project is also a potential new multifunctional tumor microenvironment regulator that can change the acidity and hypoxia of the tumor. On the one hand, it achieves sonodynamic therapy by producing ROS through sonosensitivity, thereby causing oxidative damage to tumor cells; on the other hand, its catalytic activity can produce oxygen, alleviate the hypoxia of the tumor and enhance the anti-tumor effect. Secondly, it can convert endogenous H 2 O 2It is converted into cytotoxic hydroxyl free radicals to achieve chemodynamic therapy. It also exhibits glutathione peroxidase activity, which can consume glutathione in cells and aggravate redox imbalance. In summary, copper bismuthate nanomaterials, as a heavy metal compound with sonosensitivity and catalytic activity, can fully utilize the production of hydrogen peroxide and enhance the generation of ROS in the tumor microenvironment through a stable nanoplatform and synergistic effects with other treatment methods, thereby amplifying the killing effect of oxidative damage on tumor cells and inhibiting tumor growth and spread. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] In order to make the purpose, technical solution and beneficial effects of the present invention clearer, the present invention provides the following drawings:

[0011] Figure 1 Schematic diagram of the synthesis and action of nano copper bismuthate with pH-responsive sonodynamic effect in Example 1 of the present invention.

[0012] Figure 2 This is a TEM image of nano-copper bismuthate with pH-responsive sonodynamic effect according to Example 1 of the present invention.

[0013] Figure 3 This is a graph showing the biocompatibility of the nano-copper bismuthate with pH-responsive sonodynamic effect in Example 1 of the present invention on L929 cells and HUVEC cells for 24 hours without external stimulation.

[0014] Figure 4 This is a graph showing the in vitro toxicity of the nano-copper bismuthate with pH-responsive sonodynamic effect in Example 1 of the present invention to 4T1 cancer cells for 24 h with or without ultrasonic treatment.

[0015] Figure 5 This is a diagram showing the in vivo anti-tumor effect of nano-copper bismuthate with pH-responsive sonodynamic effect in Example 1 of the present invention. DETAILED DESCRIPTION

[0016] The following examples are used to illustrate the present invention but are not intended to limit the scope of the present invention.

[0017] Example 1

[0018] (1) Preparation method of nano copper bismuthate with pH-responsive sonodynamic effect: First, 400 mg sodium methoxide and 700 mg polyvinyl pyrrolidone were added to water in a certain mass ratio to dissolve, and then 0.6 mmol 102 mg CuCl 2 ·H 2 O was ultrasonicated until a blue clear solution was obtained. 600 mg of sodium methoxide and 700 mg of polyvinyl pyrrolidone were added to water to dissolve; then 1.2 mmol 580 mg Bi(NO 3 ) 3 ·5H2 O to form an emulsion. Finally, the two liquids were mixed evenly, ultrasonically dispersed for 5 minutes, stirred at room temperature for 4 hours, and then the solution was hydrothermally treated in an oven at 140°C for 6 hours. After cooling to room temperature, centrifuged at 10,000 rpm for 2 minutes, washed with deionized water 5 times, and dried to obtain copper bismuthate nanoparticles CBO.

[0019] (2) Carboxyl modification of copper bismuthate nanoparticles: Using the EDC / NHS activation method, 1 mM CBO, 1.5 mM EDC, and 3 mM NHS were reacted at room temperature for 12 hours, and then 5.4 mg of PEG was added and stirred at room temperature for 12 hours. The excess material was removed by centrifugation to obtain nano-copper bismuthate with pH-responsive sonodynamic effect.

[0020] The transmission electron microscopy (TEM) Figure 2 ,TEM results show that the diameter of the nanoparticles is about 100 nm. Figure 3 This is the biocompatibility graph of nanoparticles to L929 cells and HUVEC cells for 24 hours without external stimulation. It can be seen that nanomedicines have good biosafety for normal cells. Figure 4 This is a graph of the in vitro toxicity of nanoparticles to 4T1 tumor cells for 24 hours without and with external stimulation. It can be seen from the graph that through different treatments, the toxicity to 4T1 tumor cells shows a good synergistic effect. Figure 5 This is a graph showing the tumor inhibition effects of different drugs in mice under different treatments. From the graph, it can be seen that compared with other groups, CBOP plus ultrasound showed a significant tumor inhibition effect.

[0021] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. A method for preparing nano copper bismuthate with pH-responsive sonodynamic effect, characterized in that: The following steps are involved: (1) Preparation method of nano-copper bismuthate with pH-responsive sonodynamic effect: First, sodium methoxide and polyvinyl pyrrolidone are added to water in a certain mass ratio to dissolve, wherein the mass ratio of sodium methoxide to polyvinyl pyrrolidone ranges from 1:1 to 1:3; then CuCl2·H2O is added and ultrasonicated until a blue clear solution is obtained. Sodium methoxide and polyvinyl pyrrolidone are added to water in a certain mass ratio to dissolve, wherein the mass ratio of sodium methoxide to polyvinyl pyrrolidone ranges from 1:1 to 1:2; then Bi(NO3)3·5H2O is added to form an emulsion. Finally, the two liquids are mixed evenly, ultrasonically dispersed for 5 minutes, stirred at room temperature for 4 hours, and then the solution is hydrothermally treated in an oven at 140°C for 6 hours. After cooling to room temperature, centrifuge at 10,000 rpm for 2 minutes, wash with deionized water 5 times, and dry to obtain copper bismuthate nanoparticles CBO; (2) Carboxyl modification of copper bismuthate nanoparticles: Using the EDC / NHS activation method, CBO / EDC / NHS were reacted at a certain molar ratio at room temperature for 12 hours, and then PEG with a mass ratio of 6:1 was added and stirred at room temperature for 12 hours. The excess material was removed by centrifugation to obtain nano-copper bismuthate CBOP with pH-responsive sonodynamic effect.

2. The method for preparing nano copper bismuthate with pH-responsive sonodynamic effect according to claim 1, characterized in that: The concentration of the CuCl2·H2O solution in step (1) is 0.6 mmol / L.

3. The method for preparing nano copper bismuthate with pH-responsive sonodynamic effect according to claim 1, characterized in that: The concentration of Bi(NO3)3·5H2O in step (1) is 1.2 mmol / L.

4. The method for preparing nano copper bismuthate with pH-responsive sonodynamic effect according to claim 1, characterized in that: The molar ratio of CBO / EDC / NHS in step (2) is 2:3:5 to 2:3:8.

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