Tumor-targeted chemokinetic therapy nanomaterials and their preparation methods and applications

Abstract

The present invention discloses a tumor-targeted chemodynamic therapy nanomaterial and its preparation method and application. Specifically, it includes mixing iron oxide, silver sulfide, ascorbic acid and extracted tumor cell membranes. After the mixed solution is ultrasonically mixed evenly, it is passed through a liposome extruder to obtain a nanomaterial with dual targeting of cell membranes and magnetic fields. The nanomaterial provided by the present invention can release AA and iron ions in the tumor microenvironment, achieve an increase in the content of hydrogen peroxide (H2O2) and photothermal-promoted chemodynamic therapy, and combine the tumor imaging capabilities of three modes: near-infrared second near-infrared (NIR-II) fluorescence imaging, magnetic resonance imaging (MRI) and photoacoustic imaging (PAI). The nanomaterial of the present invention has excellent targeting, good biocompatibility, no leakage risk, strong chemodynamic therapy characteristics and multimodal imaging functions, providing a new potential synergistic treatment method for tumor treatment.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a tumor-targeting chemodynamic therapy nanomaterial and a preparation method and application thereof. Background Art

[0002] In the past few decades, cancer has been one of the most difficult diseases in the world due to its high mortality rate. With the continuous advancement of medicine, some progress has been made in the prevention and treatment of cancer, but traditional radiotherapy, chemotherapy and other methods are often accompanied by side effects such as hair loss, loss of appetite, radiation inflammation, and low immunity. In order to solve these problems, new cancer treatment methods based on nanomedicine are constantly developing, among which chemodynamic therapy has emerged as a "green" treatment method. The mechanism of chemodynamic therapy comes from the Fenton or Fenton-like reaction, which mainly relies on the special microenvironment of the tumor to react with metal ions (such as Fe 2+ , Cu + , Mn 2+ , Pt 2+ 、V 2+ and Co 2+ ) catalyzes the conversion of excess H 2 O 2 In situ conversion of endogenous H into more toxic hydroxyl radicals (·OH) induces apoptosis or necrosis of tumor cells. It was first used in tumor treatment in 2016. Compared with other treatments based on reactive oxygen species (ROS), the advantage of chemodynamic therapy is that it does not require the introduction of external energy and can generate ·OH in situ in the tumor microenvironment without damaging normal tissues. It is considered to be a non-invasive treatment with low side effects. 2 O 2 Insufficient and weakly acidic TME limits its therapeutic effect. Although many studies have reported the application of synergistic therapy in recent years, there are still some problems such as insufficient drug loading, poor therapeutic effect, early drug leakage, and insufficient tumor specificity. Therefore, anti-tumor agents that combine two or more treatment methods to improve the therapeutic effect of cancer need to be continuously explored. In addition, visualization of tumor sites has also received increasing attention in clinical diagnosis and treatment. Traditional fluorescence imaging with visible light wavelengths often produces interference from photon scattering / absorption and tissue autofluorescence, especially when luminescent cells are deeply hidden in the tissue, which requires a higher energy excitation light source, which will also produce strong background noise, which is not conducive to tumor imaging. In contrast, fluorescence imaging in the NIR wavelength window, especially in the NIR-II region, has higher temporal / spatial resolution and penetration depth, which can greatly reduce the interference of photon scattering / absorption and tissue autofluorescence. At the same time, due to the complementary advantages of various imaging modes and the advancement of imaging methods, nanomaterials that combine multimodal imaging with tumor therapy are urgently needed.

[0003] In summary, the development of a triple-modality imaging-guided tumor dual-targeting synergistic therapeutic nanomaterial provides a new multifunctional therapeutic nanoplatform for efficient tumor treatment. Summary of the invention

[0004] In view of the shortcomings of the prior art and in order to achieve the purpose of enhancing tumor treatment, the present invention aims to provide a tumor-targeted chemodynamic therapy nanomaterial and its preparation method and application, that is, to prepare a multifunctional nanomaterial coated with 4T1 CMs, which has good tumor imaging and treatment capabilities; the prepared nanomaterial has dual targeting capabilities of CMs homologous adsorption targeting and magnetic targeting, and can accumulate at the tumor site after subcutaneous injection for precise tumor imaging and treatment.

[0005] In order to achieve the above object, the present invention adopts the following technical solution:

[0006] In a first aspect, the present invention provides a method for preparing a tumor-targeting chemodynamic therapy nanomaterial, characterized in that the nanomaterial is a composite nanomaterial, comprising the following steps:

[0007] S1: Preparation of Fe 3 O 4 and Ag 2 S nanoparticles;

[0008] S2: Fe 3 O 4 、Ag 2 S and AA solutions were mixed and ultrasonicated to obtain a uniform dispersion;

[0009] S3: Add the previously extracted CMs to the above homogenous solution and continue ultrasonic treatment until the solution is evenly dispersed;

[0010] S4: Ultrasonic mixing of the obtained mixed solution and extrusion through a liposome extruder for several cycles to obtain a composite nanomaterial with dual targeting of cell membrane and magnetic field; the particle size range of the composite nanomaterial is 90 to 400 nm.

[0011] As a preferred embodiment, in step S2, Fe 3 O 4 、Ag 2 The mass ratio of S and AA solutions is 1:(0.5~2):(0.2~1).

[0012] In the steps S2 and S3, the ultrasonic time of the mixed solution is 3 to 20 minutes, and the ultrasonic power is 300 to 600 W;

[0013] In the step S4, the mixed solution is extruded through a 200 or 400 nm polycarbonate membrane repeatedly for 5 to 10 times.

[0014] Furthermore, the Fe 3 O 4 The nanoparticles were prepared by a one-step aqueous phase method;

[0015] The water-soluble Ag 2 S quantum dots can be prepared directly in ethylene glycol phase without phase inversion;

[0016] Ag synthesized in step S1 2 The fluorescence emission peak of S is in NIR-II, 1000-1700nm, and it shows greater advantages in in vivo imaging due to its low toxicity, deep penetration, and low background interference;

[0017] The tumor CMs are derived from mouse tumor cells, including 4T1 breast cancer cells, and are extracted using a cell membrane extraction kit.

[0018] In a second aspect, the present invention provides a tumor-targeting chemodynamic therapy nanomaterial, characterized in that the nanomaterial is prepared by any of the preparation methods described above.

[0019] In a third aspect, the present invention provides a use of the above-mentioned nanomaterial in the preparation of tumor therapeutic drugs.

[0020] As a preferred embodiment, the application is specifically that the tumor-targeted chemodynamic therapy nanomaterial is verified in a 4T1 tumor-bearing mouse experiment, and the experiment includes tumor imaging and tumor treatment.

[0021] Furthermore, the drug is an anti-tumor nanomedicine guided by three imaging modes for clarified dual-targeted and highly efficient chemodynamic therapy; the obtained tumor therapeutic drug has the ability to perform NIR-II fluorescence imaging, MRI and PA imaging at the tumor site of mice.

[0022] Furthermore, the tumor treatment application was first verified by cell experiments, and the cells included: 4T1 cells, Hela cells, COS7 cells, and HEK293T cells.

[0023] Furthermore, the administration method of the drug for treating mouse tumors is selected from subcutaneous injection and an external magnetic field for magnet targeted attraction. It has a synergistic treatment effect of photothermal and chemodynamics, can effectively eliminate tumors and has no obvious toxic side effects.

[0024] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0025] (1) The nanomaterials prepared by the present invention not only avoid premature leakage of the materials due to the presence of CMs, but also improve the homologous targeting of the nanomaterials. 3 O 4The presence of gives the composite nanomaterials excellent magnetic targeting ability. Compared with single cell membrane targeting, this dual targeting ability has the advantages of being more precise and efficient.

[0026] (2) The nanomaterials prepared by the present invention simultaneously deliver Fe to the tumor site 3 O 4 and AA, which is catalyzed by metal ions at the tumor site to produce H 2 O 2 , providing sufficient raw materials for the iron-mediated Fenton reaction. At the same time, AA has no damage to normal cells and avoids the introduction of exogenous H 2 O 2 or glucose oxidase due to leakage risk.

[0027] (3) The nanomaterial prepared by the present invention has good photothermal capacity, with a photothermal efficiency of 55.6%, and can effectively convert 808nm laser into heat to promote the chemodynamic treatment effect.

[0028] (4) The nanomaterials prepared by the present invention have the capabilities of NIR-II fluorescence imaging, PA imaging and MRI. The complementary advantages of the three-mode imaging enable tumor imaging to more clearly display the tumor location, and have good application prospects in tumor diagnosis and treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Schematic diagram of the tumor treatment mechanism described in Example 1.

[0030] Figure 2 This is the transmission electron microscopy image of the FAA@CM nanomaterial in Example 1.

[0031] Figure 3 This is the Zeta potential diagram of different materials in Example 1.

[0032] Figure 4 This is a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) image of different materials in Example 1.

[0033] Figure 5 is Fe in Example 1 3 O 4 and X-ray diffraction (XRD) patterns of FAA@CM nanomaterials.

[0034] Figure 6 This is the element mapping image of the FAA@CM nanomaterial in Example 1.

[0035] Figure 7 4 is the Fourier infrared spectra of different materials in Example 1.

[0036] Figure 8This is the X-ray photoelectron spectroscopy (XPS) diagram of the FAA@CM nanomaterial in Example 1.

[0037] Fig. 9 This is the NIR-II fluorescence emission spectrum of the FAA@CM nanomaterial in Example 1.

[0038] Fig.10 The FAA@CM nanomaterial in Example 2 was exposed to 100 μM H 2 O 2 The UV absorption spectrum of methylene blue (MB) at 665nm was reduced in the presence of .

[0039] Fig.11 The FAA@CM nanomaterial in Example 2 was exposed to 100 μM H 2 O 2 The fluorescence spectrum of terephthalic acid (TA) in the presence of 2-nitropropene shows a fluorescence peak at 435 nm.

[0040] Fig.12 The solution of different nanomaterials or the separate H in Example 2 2 O 2 Electron spin resonance (ESR) spectroscopy.

[0041] Fig.13 The temperature rise curves of FAA@CM solutions and water with different concentrations in Example 3.

[0042] Fig.14 The heating and cooling curves of the FAA@CM solution in Example 3 (curve), and the linear fit of the time of the natural cooling period and the negative natural logarithm of the driving force temperature of FAA@CM (straight line).

[0043] Fig.15 The temperature change of the FAA@CM solution in Example 3 after three irradiation / cooling cycles.

[0044] Fig.16 This is the NIR-II fluorescence image of the FAA@CM solution in Example 4.

[0045] Fig.17 PA signals of FAA@CM solutions with different concentrations in Example 4.

[0046] Fig.18 is the longitudinal relaxation (r) of FAA@CM solutions with different concentrations in Example 4 1 ) value and the corresponding T 1 Weighted MRI images.

[0047] Fig.19 is the transverse relaxation (r) of FAA@CM solutions with different concentrations in Example 4 2) value and the corresponding T 2 Weighted MRI images.

[0048] Fig. 20 is the Fe in Example 5 with or without 808nm laser irradiation. 3 O 4 and FAA@CM incubation on the cytotoxicity of 4T1 cells.

[0049] Fig.21 The effect of FAA@CM in Example 5 on the cell viability of MCF-7 cells, Hela cells, COS7 cells and HEK293T cells.

[0050] Fig. 22 This is the flow cytometric analysis of 4T1 cells under different treatments in Example 5.

[0051] Fig.23 This is a confocal laser scanning microscopy (CLSM) image of 4T1 cells co-stained with calcein-AM and propidium iodide (PI) in Example 5. Scale bar: 100 μm.

[0052] Fig.24 These are NIR-II fluorescence images of 4T1 tumor-bearing mice at different times after subcutaneous injection of FAA@CM in Example 6.

[0053] Fig.25 is the T of 4T1 tumor-bearing mice at different times after subcutaneous injection of FAA@CM in Example 6 1 / T 2 Weighted MR images.

[0054] Fig.26 PA images of the tumor site of 4T1 tumor-bearing mice at different times after subcutaneous injection of FAA@CM in Example 6.

[0055] Fig. 27 This is the relative tumor volume growth curve of mice in Example 7.

[0056] Fig.28 These are photos of tumors in tumor-bearing mice after 16 days of different treatments in Example 7.

[0057] Fig.29 This is the body weight change curve of mice treated with different methods within 16 days in Example 7.

[0058] Fig.30 These are the liver and kidney function test results of different groups of mice after 16 days of treatment in Example 7.

[0059] Fig.31 These are the results of routine blood tests of mice in different groups after 16 days of treatment in Example 7.

[0060] Fig.31 middle: a) The recorded items were white blood cell count (WBC), lymphocyte count (Lymph), monocyte count (Mon), neutrophil count (Gran), red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration (MCHC), coefficient of variation of red cell distribution width (RDW), platelet count (PLT) and mean platelet volume (MPV). DETAILED DESCRIPTION

[0061] The technical solution of the present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0062] The present invention provides a tumor-targeting chemodynamic therapy nanomaterial and a preparation method thereof. The nanomaterial of the present invention will be described in detail below in conjunction with examples and experimental data.

[0063] See also Figure 1 , Figure 1 Schematic diagram of the treatment and imaging principle of the tumor-targeting chemodynamic therapeutic nanomaterial provided by the present invention for 4T1 tumor-bearing mice.

[0064] like Figure 1 It can be seen that the prepared nanomaterials were subcutaneously injected into tumor-bearing mice, and accumulated at the tumor site through the dual targeting effects of CMs homologous adsorption and magnetic field guidance, and were internalized by the tumor, thereby releasing Fe 3 O 4 、Ag 2 S and AA. AA plays a pro-oxidative role under the catalysis of iron ions, increasing H 2 O 2 Content, increased H 2 O 2 The level will amplify the efficiency of Fenton reaction-based CDT ablation of tumors, and jointly enhance the effect of tumor chemodynamic therapy under the promotion of photothermal effect. In addition, FAA@CM is an excellent tumor imaging agent that can provide three imaging modes: NIR-II fluorescence imaging, MRI imaging, and PA imaging, which can be applied to tumor visualization. The results of in vivo and in vitro experiments showed that compared with single CDT treatment, the therapeutic effect of FAA@CM+NIR+MF was significantly enhanced, indicating the feasibility of this synergistic catalytic therapeutic agent in tumor treatment. (MF stands for magnetic field)

[0065] Example 1 Tumor-targeted chemodynamic therapy nanomaterial and preparation method thereof

[0066] The preparation method of the above-mentioned nanomaterial comprises the following steps:

[0067] (1)Fe 3 O 4 Synthesis

[0068] The ferrous ions are precipitated under strong alkaline conditions, and then polyethyleneimine is added as a stabilizer, and the temperature is raised and refluxed for 2 hours to obtain the product. The specific synthesis steps are as follows:

[0069] Weigh 0.128g FeSO 4 7H 2 O was dissolved in 8 ml of ultrapure water (deoxygenated with argon), and then 2.0 M KNO 3 and 1.0M NaOH solution, take 1 ml of each and add into the FeSO 4 The solution was added with 40 mg PEI and the solution turned from green to black. The reaction temperature was raised to 90 °C and refluxed for 2 h to finally obtain PEI-coated Fe 3 O 4 The reaction solution was separated by a magnet, and the product was finally resuspended in ultrapure water to obtain Fe 3 O 4 of aqueous solution.

[0070] (2) Ag 2 Synthesis of S

[0071] 10 mL of ethylene glycol (EG) was added to a 50 mL three-necked flask. After passing argon for a while, the solution was heated to 110 °C, 100 μL of mercaptopropionic acid was added, and then 0.05 mmol of AgNO was added. 3 , continue to heat to 145℃ and maintain the reaction at this temperature. After a certain period of reaction, stop stirring and heating, cool the reaction solution to room temperature, and the precipitate obtained after centrifugation is the obtained Ag 2 S quantum dots can be directly dispersed in aqueous solution.

[0072] (3) Synthesis of FAA@CM

[0073] The Fe synthesized above 3 O 4 、Ag 2 S and the prepared AA solution were mixed, ultrasonically treated to obtain a uniform dispersion, and then the extracted CMs were added, and ultrasonic treatment was continued until the solution was uniformly dispersed; finally, the above solution was extruded through a liposome extruder for several cycles to obtain a clarified dual-targeted and highly efficient chemodynamic therapy anti-tumor nanodrug FAA@CM guided by three imaging modes.

[0074] Characterization and performance:

[0075] Transmission electron microscopy images Figure 2It was shown that FAA@CM was monodispersed spherical nanoparticles with an average particle size of 81.6±2.6nm. Figure 3 The change in zeta potential indicated that the nanoparticles were successfully coated by the cancer cell membrane. Figure 4 The SDS-PAGE results were used to analyze cell membrane proteins, and the results showed that FAA@CM nanomaterials had similar proteins to 4T1 cell membranes, proving the successful coating of cell membranes on the nanomaterials. Figure 5 The XRD pattern of Fe 3 O 4 and Ag 2 The existence of S. Figure 6 Element mapping, Figure 7 The infrared spectrum and Figure 8 The XPS results show the coexistence of Fe, Ag and P elements. Fig. 9 The NIR-II fluorescence spectrum shows that the synthesized nanomaterials have the ability of NIR-II fluorescence imaging.

[0076] Example 2 Catalytic performance of nanomaterials for targeted tumor chemodynamic therapy

[0077] To evaluate the generation of ·OH, FAA@CM and H 2 O 2 (100 μM) were co-incubated in acetic acid-sodium acetate buffer at pH 6.5, and the generation of ·OH was detected by the changes in the UV absorption curve and fluorescence emission spectrum of MB (0.025 mM) and TA (1 mM), respectively. 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) was used as the capture agent in the ESR test.

[0078] Characterization and performance:

[0079] like Fig.10 As shown in Figure 2, the UV spectral intensity absorption of MB decreases with time, while that of FAA@CM or H 2 O 2 The reduction was not significant. Fig.11 The time-varying spectrum of TAOH in the solution also demonstrates the ability of the nanoparticles to generate strong reactive oxygen species. Fig.12 In the ESR test spectrum, the FAA@CM group showed a 3 O 4 The stronger characteristic 1:2:2:1 electron spin signal, indicating the stronger production of ·OH under the condition of AA-promoted oxidation, demonstrated the enhanced chemokinetic therapeutic properties of the nanomaterials.

[0080] Example 3 Photothermal performance of nanomaterials for targeted tumor chemodynamic therapy

[0081] Different concentrations of FAA@CM nanomaterials were dispersed in water and irradiated with 808 nm laser (1.0 W cm -2 ). The temperature change of the nanomaterial during irradiation was measured using an infrared thermal imager. The photothermal conversion efficiency (η) was calculated according to equation (1):

[0082]

[0083] where m is the mass of the solution, which is equal to 1.0 g in the current study, c is the heat capacity of water, which is equal to 4.2 J / g, and T max and T max , H2O is the maximum temperature of the nanomaterial solution and water during irradiation. I is the power density of the laser, which is equal to 1 W cm in the current study. -2 ), A is the absorbance of the nanomaterial solution at 808nm, which can be measured by ultraviolet spectrometer. Fig.14 The linear regression of the cooling curve in the system gives the system time constant τ s For in vivo photothermal imaging, PBS and FAA@CM nanomaterials (5 mg kg -1 ) were injected into anesthetized 4T1 tumor-bearing mice. The temperature of the tumor surface was measured using an infrared thermal imager 4 hours after injection.

[0084] Characterization and performance:

[0085] Fig.13 The temperature rise curves show that under 808nm laser irradiation, the temperature rise of solutions of various concentrations shows a significant concentration-dependent photothermal effect. Fig.14 The results show that the photothermal conversion efficiency of FAA@CM is 55.6%. Fig.15 It shows that FAA@CM still maintains good photothermal stability after three heating and cooling cycles. The above results prove that FAA@CM is expected to become a good photothermal therapeutic agent.

[0086] Example 4 In vitro multimodal imaging of nanomaterials for chemodynamic therapy targeting tumors

[0087] For in vitro NIR-II imaging, the FAA@CM solution was placed in a centrifuge tube and tested using a near-infrared fluorescence imager. For in vitro PA imaging, the FAA@CM aqueous solution was diluted to different concentrations and used for PA imaging. For in vitro MRI imaging, the MRI images and t of the FAA@CM aqueous solution were measured using a Siemens Magnetom Prisma 3.0T magnetic resonance MRI system. 1 / t 2 Relaxation time.

[0088] Characterization and performance:

[0089] Fig.16 Shows that FAA@CM retains Ag 2 The NIR-II fluorescence imaging capability of S is expected to provide fluorescence imaging guidance in vitro and in vivo. Fig.17 As shown in Figure 3, the PA imaging intensity shows a significant concentration dependence. As the solution concentration increases, the PA in vitro imaging signal increases linearly, demonstrating that FAA@CM has good PA imaging performance. Fig.18 and Fig.19 The MRI results of FAA@CM show that the r 1 and r 2 Relaxation rate value. As the concentration of FAA@CM increases, the T 1 The image becomes brighter, T 2 The image becomes darker. 1 and r 2 The values ​​were 4.85 and 50.52 mM s -1 Based on the experimental results and discussions of the present invention, FAA@CM has the potential to be a triple-modal imaging agent for tumor diagnosis and treatment.

[0090] Example 5 Cytotoxicity Experiment of Nanomaterials for Chemodynamic Therapy of Tumor Targeting

[0091] HEK293T cells, COS7 cells, Hela cells, MCF-7 cells and 4T1 cells were seeded in 96-well plates (5 × 10 3 Cells / well) were cultured for 12 hours. Then different concentrations of FAA@CM nanoparticles were added to the culture medium. After 24 hours of co-incubation, the relative cell viability was measured by MTT assay. The potential cell death mechanism was determined by flow cytometry Annexin V-FITC / PI double staining analysis. 2,7-Dichlorodihydrofluorescein diacetate (DCFH-DA) was used as a ROS probe to monitor the ROS level in the cells, which was more intuitively characterized by confocal laser scanning microscopy.

[0092] Characterization and performance:

[0093] Fig. 20 The MTT results of 4T1 cells showed that the nanoparticles significantly reduced the percentage of viable 4T1 cells in a concentration-dependent manner. At the same concentration, FAA@CM showed a higher 3 O4 is much more cytotoxic, which may be due to the oxidative formation of H 2 O 2 The Fenton reaction is promoted. Fig.21It was shown that FAA@CM had no obvious cytotoxicity in other tumor cells (MCF-7, Hela cells) and normal cells (COS 7 cells, HEK293T cells). This was due to the homologous adhesion of the cell membrane coating, which resulted in a lower uptake of FAA@CM by other cells. The above results demonstrated that FAA@CM was highly efficient and specific in killing cancer cells. Fig. 22 The results showed that apoptosis is the main way leading to cell death. Fig.23 The results showed that cells cultured with FAA@CM nanoparticles showed stronger fluorescence, indicating that more ROS were generated. These results all proved that FAA@CM nanoparticles had stronger cytotoxicity against 4T1 cancer cells due to their selectivity and enhanced Fenton effect.

[0094] Example 6 In vivo multimodal imaging of nanomaterials for chemodynamic therapy targeting tumors

[0095] Mice bearing 4T1 tumors were anesthetized and FAA@CM nanomaterials (dose: 5 mg kg -1 ) was injected subcutaneously into mice. Images of the mouse body or tumor site before and after injection were recorded at designated times using the NIR-II fluorescent in vivo imaging system, PA images, and MRI scanners.

[0096] Characterization and performance:

[0097] Fig.24 The fluorescence images showed that the fluorescence intensity at the tumor location increased over time and reached a maximum value 4 h after injection, indicating that the nanoparticles were efficiently accumulated due to the homologous adhesion effect of the tumor cell membrane. And due to the targeting of the magnetic field, FAA@CM+MF can significantly enhance the accumulation of nanoparticles in tumors. Fig.25 and Fig.26 The MRI and PA imaging results showed that the dual-targeted FAA@CM nanomaterials have effective tumor enrichment and imaging capabilities. Therefore, tumor microenvironment-specific FAA@CM has great potential to achieve precise drug delivery and imaging.

[0098] Example 7 In vivo targeted therapeutic effect of tumor-targeted chemodynamic therapy nanomaterials

[0099] Female BALB / c mice were purchased from Wuhan Experimental Animal Center and treated according to the protocol approved by Wuhan University Experimental Animal Center. 4T1 cell suspension (100 μL, 1×10 6 cells) were injected subcutaneously into the back of 5-week-old mice. When the tumor volume of the mice reached 100mm 3The in vivo experiment was started at 1:10 pm. All mice received subcutaneous injections of different agents every three days. The magnet was placed at the tumor site of the FAA@CM+MF+NIR experimental group for 4 h, while the mice in the other groups were not exposed to the magnetic field. The weight and tumor size of the mice in each treatment group were recorded every two days. The tumor volume was obtained using equation (2):

[0100] Volume = length × (width) 2 / twenty two)

[0101] After 16 days of treatment, all mice were killed, and tumors were collected to record tumor volumes and take photos. Whole blood was collected from healthy mice and experimental mice for liver and kidney function and blood routine analysis.

[0102] Characterization and performance:

[0103] Fig. 27 The relative tumor volume growth curve and Fig.28 The tumor images showed that the tumor size of mice injected with PBS increased rapidly regardless of whether they were irradiated with laser or not, while in other treatment groups, photothermal promotion or photothermal plus AA-promoted chemodynamic therapy had better tumor inhibition effects. In the FAA@CM+MF+NIR group, some tumors were completely eliminated due to dual targeting and enhanced therapeutic effects, demonstrating the excellent tumor targeting and therapeutic ability of FAA@CM.

[0104] Most importantly, during treatment, Fig.29 There was no significant change in the body weight of the mice, suggesting that the treatment had negligible side effects on the body. Fig.30 and Fig.31 All preparations showed no significant hepatotoxicity, nephrotoxicity, and hematotoxicity. These results confirmed the good biocompatibility of FAA@CM and its advanced dual-targeting and laser-assisted tumor treatment effects.

[0105] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0106] The embodiment of the present invention provides a tumor-targeting chemokinetic therapy nanomaterial and its preparation method and application. The present invention designs to use 4T1 tumor CMs to coat Fe 3 O 4 、Ag 2 S and AA are three components to synthesize composite nanomaterials, which not only introduces AA to increase the local H 2 O 2 content, and at the same time used the photothermal properties of nanomaterials to synergistically enhance chemodynamic therapy, showing good tumor inhibition effects in cell experiments and in vivo experiments.

[0107] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing a tumor-targeting chemodynamic therapy nanomaterial. Features: The nano material is a composite nano material, comprising the following steps: S1: Preparation of Fe 3 O 4 and Ag 2 S nanoparticles; precipitate ferrous ions under strong alkaline conditions, then add polyethyleneimine as a stabilizer, and reflux at elevated temperature for 2 hours to obtain Fe 3 O 4 Nanoparticles; the Ag 2 S nanoparticles are water-soluble Ag 2 S quantum dots, Ag 2 S quantum dots are directly prepared in the ethylene glycol phase without phase inversion; S2: Fe 3 O 4 、Ag 2 S and AA solutions are mixed and ultrasonically treated to obtain a uniform dispersion; in step S2, Fe 3 O 4 、Ag 2 The mass ratio of S and AA solutions is 1:(0.5 ~ 2):(0.2 ~ 1); S3: adding the previously extracted CMs to the uniform dispersion obtained above, and continuing ultrasonic treatment until the solution is uniformly dispersed; in the steps S2 and S3, the ultrasonic time of the mixed solution is 3 to 20 minutes, and the ultrasonic power is 300 to 600W; S4: ultrasonically mixing the obtained mixed solution and extruding it through a liposome extruder for several cycles; obtaining a composite nanomaterial with dual targeting of cell membrane and magnetic field; the particle size range of the composite nanomaterial is 90 to 400 nm; in the step S4, the mixed solution is repeatedly extruded through a 200 or 400 nm polycarbonate membrane for 5 to 10 times; The Ag synthesized in step S1 2 The fluorescence emission peak of S is in the NIR-II region, with a wavelength range of 1000-1700 nm. It has greater advantages in in vivo imaging due to its low toxicity, deep penetration, and low background interference; The tumor CMs are derived from mouse tumor cells, including 4T1 breast cancer cells, and are extracted using a cell membrane extraction kit.

2. A tumor-targeted chemodynamic therapy nanomaterial, Features: The nanomaterial is prepared by the preparation method as claimed in claim 1.

3. Use of the nanomaterial as claimed in claim 2 in the preparation of tumor therapeutic drugs, Features: The application is specifically that the tumor-targeted chemodynamic therapy nanomaterial is verified in a 4T1 tumor-bearing mouse experiment, and the experiment includes tumor imaging and tumor treatment; The drug is an anti-tumor nanomedicine guided by three imaging modes for clarified dual-targeted and highly efficient chemodynamic therapy; the obtained tumor therapeutic drug has the ability to perform NIR-II fluorescence imaging, MRI and PA imaging at the tumor site of mice; The tumor treatment application was first verified by cell experiments, and the cells included: 4T1 cells, Hela cells, COS7 cells, and HEK293T cells; The administration method of the drug for treating mouse tumors is selected from subcutaneous injection and an external magnetic field for magnet targeted attraction. It has a synergistic treatment effect of photothermal and chemodynamics, can effectively eliminate tumors and has no obvious toxic side effects.

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