Acoustic / chemical power synergistic anti-tumor nano-drug as well as preparation method and application thereof

Abstract

The invention relates to the technical field of nano-drugs, in particular to an acoustic / chemical power synergistic anti-tumor nano-drug as well as a preparation method and application thereof. The invention provides an acoustic / chemical power synergistic anti-tumor nano-drug, which takes hollow mesoporous MnO2 nano-particles as a carrier, and is loaded with vitamin C and a sound-sensitive agent chlorin e6. Vc can generate abundant H2O2 to provide a raw material for CDT, and MnO2 is used as a peroxidase-like enzyme to catalyze H2O2 to generate O2, so that tumor hypoxia is effectively relieved, and the effect of sonodynamic therapy is enhanced. Besides, GSH induces MnO2 degradation, GSH consumption is caused to enhance CDT, GSH depletion can cause glutathione peroxidase 4 (GPX4) activity reduction, ROS generation and GPX4 activity reduction have a synergistic effect, lipid peroxides (LPOs) are increased, and finally ferroptosis of tumor cells is caused.

Description

Technical Field

[0001] The present invention relates to the technical field of nanomedicine, and in particular to a sonic / chemical dynamic synergistic anti-tumor nanomedicine and a preparation method and application thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Malignant tumors have become one of the major public issues that seriously threaten people's health. Currently, traditional treatments for malignant tumors mainly include surgery, radiotherapy, chemotherapy, and immunotherapy, but all of them have certain limitations. Therefore, non-invasive, highly targeted, and low / no toxicity treatment strategies are urgently needed.

[0004] Sonodynamic therapy (SDT) is a new non-invasive treatment method that has emerged in recent years. It is based on photodynamic therapy. Unlike the latter, SDT uses ultrasound instead of laser as the source of therapeutic energy, overcoming the limitations of laser penetration depth and skin phototoxicity associated with traditional photosensitizers. At present, the possible mechanisms of SDT in treating tumors mainly include reactive oxygen theory, ultrasonic cavitation effect and thermal damage. (1) Reactive oxygen theory: Ultrasound excites sonosensitizers to produce singlet oxygen with biological toxicity, which increases the level of reactive oxygen inside tumor cells and causes irreversible damage to tumor cells / tissues; (2) Ultrasonic cavitation effect: When ultrasound acts on the surface of the cell membrane, the sound energy generated by ultrasound can rapidly increase mechanical pressure and promote the generation of microbubbles in the tissue fluid. The microbubbles around the cells generate mechanical effects through their oscillations to instantly form micropores on the cell membrane, thereby increasing vascular permeability and drug transport capacity; (3) Thermal damage: The absorption and conversion of ultrasonic mechanical energy induces the generation of thermal energy during its propagation, which increases the temperature of the tissue and leads to the death of cancer cells. As a non-invasive therapeutic strategy for targeted killing of tumor cells, SDT uses low-intensity ultrasound to activate sonosensitizers, thereby mediating the production of 1 O 2 To kill tumor cells.

[0005] Different from SDT, chemodynamic therapy (CDT) is a method that utilizes Fenton or Fenton-like reactions to catalyze the release of hydrogen peroxide (H 2 O 2) is a therapeutic strategy that produces highly cytotoxic hydroxyl radicals in situ to kill cancer cells. CDT can occur autonomously and does not rely on external energy intervention. However, the endogenous reaction substrates of tumors are limited, which restricts the effective implementation of CDT. When combined with SDT, the deep diffusion of nanomedicines and the expanded production of reactive oxygen species can enhance the therapeutic effect on tumors. In addition, previous research results have also shown that the introduction of ultrasound can enhance the activity of the Fenton reaction and thereby amplify the efficiency of CDT. Therefore, SDT synergizes with CDT to improve treatment efficiency, which is of great significance and has broad clinical application prospects. Summary of the invention

[0006] In order to overcome the above problems, the present invention provides a sono / chemodynamic synergistic anti-tumor nanomedicine and a preparation method and application thereof.

[0007] In order to achieve the above technical objectives, the present invention adopts the following technical solutions:

[0008] The first aspect of the present invention provides a sono / chemical synergistic anti-tumor nanomedicine, wherein the nanomedicine is hollow mesoporous MnO 2 Nanoparticles are used as carriers to load vitamin C (Vc) and sonosensitizer dihydrochlorin e6 (Ce6).

[0009] The second aspect of the present invention provides a method for preparing the above-mentioned sono / chemical dynamic synergistic anti-tumor nanomedicine, comprising:

[0010] (1) Silica nanoparticles were prepared using tetraethyl orthosilicate as raw material, and KMnO was added to the silica nanoparticle solution. 4 Solution, obtain SiO 2 @MnO 2 Nanoparticles, SiO 2 @MnO 2 Nanoparticles as templates to remove SiO 2 , to obtain hollow mesoporous MnO 2 Nanospheres;

[0011] (2) Hollow mesoporous MnO 2 The nanospheres were dispersed in the vitamin C solution and stirred in the dark to obtain the nano drug intermediate MnO 2 -Vc;

[0012] (3) The sonosensitizer dihydrochlorin e6 was modified with polyethylene glycol to obtain mPEG-Ce6, and mPEG-Ce6 was reacted with the nanodrug intermediate MnO 2 -Vc after mixing, stir and react in dark to obtain nano drug MnO 2 -Vc@mPEG-Ce6.

[0013] The third aspect of the present invention provides the use of the above-mentioned sono / chemical synergistic anti-tumor nanomedicine in the preparation of anti-tumor drugs.

[0014] The fourth aspect of the present invention provides an anti-tumor drug composition, comprising the above-mentioned sono / chemodynamic synergistic anti-tumor nanodrug.

[0015] The beneficial effects of the present invention are:

[0016] (1) The present invention provides a sono / chemical synergistic anti-tumor nanomedicine, which is hollow mesoporous MnO 2 Nanoparticles are used as carriers to load vitamin C and sonosensitizer dihydrochlorin e6. The tumor microenvironment is usually characterized by hypoxia, relative lack of hydrogen peroxide and excess glutathione (GSH), which accelerate the progression and metastasis of tumor cells. The nanomedicine provided by the present invention can play a catalytic function in turn according to the specific tumor microenvironment to improve the efficiency of ROS generation. Specifically, Vc can produce abundant H 2 O 2 , providing raw materials for CDT, while MnO 2 As a peroxidase, it catalyzes H 2 O 2 Produce O 2 , thereby effectively alleviating tumor hypoxia and enhancing the effect of sonodynamic therapy. In addition, GSH induces MnO 2 Degradation leads to GSH consumption and enhanced CDT. GSH depletion leads to reduced activity of glutathione peroxidase 4 (GPX4). The production of ROS and the reduction of GPX4 activity work together to increase lipid peroxides (LPOs), ultimately leading to ferroptosis of tumor cells. Therefore, the nanomedicine provided by the present invention can make sonodynamics and chemical dynamics synergistically anti-tumor, induce ferroptosis of tumor cells, and then inhibit tumor growth.

[0017] (2) Compared with other H 2 O 2 The precursor substance, Vc, has good water solubility and is therefore more easily absorbed by the body, thereby increasing its bioavailability. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0019] Figure 1 The hollow mesoporous MnO in Example 1 2 Particle size (a) and pore size (b) of the sphere;

[0020] Figure 2Nano drug MnO 2 - Transmission electron microscopy (TEM) image of Vc@mPEG-Ce6;

[0021] Figure 3 Nanomedicine MnO under different concentrations (A) and different pH conditions (B) 2 -Vc@mPEG-Ce6 catalyzes H 2 O 2 Decomposition produces O 2 A graph showing the change of the amount of over time;

[0022] Figure 4 The ESR signals of ·OH generated by different treatment groups were detected by using DMPO as a trapping agent (A). The ESR signals of ·OH generated by different treatment groups were detected by using DMPO as a trapping agent (B). 1 O 2 Generated ESR signal (B);

[0023] Figure 5 Different concentrations of nano drug MnO 2 -Consumption of GSH (1 mM) by Vc@mPEG-Ce6;

[0024] Figure 6 The effect of nano drug MnO on tumor cells (PC-3 cells) 2 -Vc@mPEG-Ce6 uptake and phagocytosis test results;

[0025] Figure 7 The cell killing ability experiment of tumor cells (PC-3) under different experimental conditions; from left to right: 1) control group; 2) MnO 2 (200ppm) group; 3) Ce6 (200ppm) + ultrasound (1w / cm 2 ,3min) group; 4)MnO 2 -Vc@mPEG-Ce6 (200ppm) group; 5) MnO 2 -Vc@mPEG-Ce6(200ppm)+ultrasound(1w / cm 2 , 3min) group;

[0026] Figure 8 For intravenous injection of MnO 2 -Fluorescence images of tumor-bearing mice at different time points after Vc@mPEG-Ce6 injection (A) and in vitro photographs and fluorescence images of the heart, liver, spleen, lung, kidney and tumor of mice 24 hours after intravenous injection (B);

[0027] Fig. 9 Tumor-bearing mice were injected with MnO via the tail vein 2-Vc@mPEG-Ce6, and the operation diagram of ultrasound treatment within 24 hours (A), the curve diagram of the changes of mouse weight (B) and tumor volume (C) after treatment with different methods over time;

[0028] Fig.10 The GPX4 immunohistochemistry images of mouse tumor sections treated with different methods are shown in Figure 1. From left to right, they are: 1) control group; 2) MnO 2 (200ppm) group; 3) Ce6 (200ppm) + ultrasound (1w / cm 2 ,3min) group; 4)MnO 2 -Vc@mPEG-Ce6 (200ppm) group; 5) MnO 2 -Vc@mPEG-Ce6(200ppm)+ultrasound(1w / cm 2 , 3min) group results. DETAILED DESCRIPTION

[0029] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0030] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0031] The first typical embodiment of the present invention provides a sono / chemical synergistic anti-tumor nanomedicine, wherein the nanomedicine is hollow mesoporous MnO 2 Nanoparticles are used as carriers to load vitamin C (Vc) and sonosensitizer dihydrochlorin e6 (Ce6).

[0032] In one or more embodiments, the MnO 2 Nanoparticles are MnO 2 Nanospheres;

[0033] Preferably, MnO 2 The particle size of the nanospheres is about 190nm; the pore size is about 3.9nm.

[0034] A second typical embodiment of the present invention provides a method for preparing the above-mentioned sono / chemical dynamic synergistic anti-tumor nanomedicine, comprising:

[0035] (1) Silica nanoparticles were prepared using tetraethyl orthosilicate as raw material, and KMnO was added to the silica nanoparticle solution. 4 Solution, obtain SiO 2 @MnO 2 Nanoparticles, SiO 2 @MnO 2 Nanoparticles as templates to remove SiO 2 , to obtain hollow mesoporous MnO 2 Nanospheres;

[0036] (2) Hollow mesoporous MnO 2 The nanospheres were dispersed in the vitamin C solution and stirred in the dark to obtain the nano drug intermediate MnO 2 -Vc;

[0037] (3) The sonosensitizer dihydrochlorin Ce6 was modified with polyethylene glycol to obtain mPEG-Ce6, and mPEG-Ce6 was reacted with the nanodrug intermediate MnO 2 -Vc after mixing, stir and react in dark to obtain nano drug MnO 2 -Vc@mPEG-Ce6.

[0038] In one or more embodiments, in step (1), the silica nanoparticles and KMnO 4 The molar ratio is 2.5 to 3:1, preferably 2.85:1.

[0039] In one or more embodiments, in step (1), KMnO 4 The concentration of the solution is 0.015 to 0.03 mmol / L, preferably 0.02 mmol / L.

[0040] In one or more embodiments, in step (1), ultrasonically obtain SiO 2 @MnO 2 For nanoparticles, the ultrasonic power is 140-160W, preferably 150W; the time is 5-7h, preferably 6h.

[0041] In one or more embodiments, in step (1), Na 2 CO 3 Etching to remove SiO 2 , Na 2 CO 3 The concentration is 0.015-0.03 mmol / L, preferably 0.02 mmol / L; the etching temperature is 55-65°C, preferably 60°C; the etching time is 10-15h, preferably 12h; the stirring rate during etching is 120-200rpm, preferably 150rpm.

[0042] In one or more embodiments, in step (2), the hollow mesoporous MnO 2 The mass ratio of nanospheres to vitamin C is 0.03-0.01:1, preferably 0.05:1.

[0043] In one or more embodiments, in step (2), the concentration of vitamin C in the vitamin C solution is 1 to 3 g / L, preferably 2 g / L.

[0044] In one or more embodiments, in step (2), the temperature of the reaction under stirring in the dark is room temperature, and the stirring time is 10 to 15 hours, preferably 12 hours.

[0045] In one or more embodiments, in step (3), Ce6 and MnO 2 The mass ratio is 8 to 12:1, preferably 10:1.

[0046] In one or more embodiments, in step (3), the temperature of the reaction under stirring in the dark is room temperature, and the stirring time is 20 to 30 hours, preferably 24 hours.

[0047] A third typical embodiment of the present invention provides the use of the above-mentioned sono / chemical synergistic anti-tumor nanomedicine in the preparation of anti-tumor drugs.

[0048] A fourth typical embodiment of the present invention provides an anti-tumor drug composition, comprising the above-mentioned sono / chemodynamic synergistic anti-tumor nanodrug.

[0049] In one or more embodiments, the anti-tumor drug composition also includes drugs related to tumor treatment.

[0050] Preferably, the tumor treatment-related drug is an anti-tumor drug, an immune adjuvant, a checkpoint inhibitor or an antigen protein.

[0051] In one or more embodiments, the tumor includes benign tumors and / or malignant tumors; the malignant tumors include solid tumors and hematological tumors.

[0052] It should be noted that tumor is used in the present invention as known to those skilled in the art, and includes benign tumors and / or malignant tumors. Benign tumors are defined as excessive proliferation of cells that cannot form aggressive, metastatic tumors in the body. Conversely, malignant tumors are defined as cells with multiple cellular abnormalities and biochemical abnormalities that can form systemic diseases (e.g., tumor metastasis in distant organs).

[0053] Solid tumors such as breast, bladder, bone, brain, central and peripheral nervous system, colon, endocrine glands (such as thyroid and adrenal cortex), esophagus, endometrium, germ cells, head and neck, kidney, liver, lung, larynx and hypopharynx tumors, mesothelioma, ovary, pancreas, prostate, rectum, kidney, small intestine, soft tissue, testicle, stomach, skin (such as melanoma), ureter, vagina and vulva tumor. Malignant tumors include hereditary cancers, such as retinoblastoma and Wilms tumor (Wilms tumor). In addition, malignant tumors include primary tumors in the organs and corresponding secondary tumors (tumor metastasis) in distal organs. Hematological tumors such as aggressive and indolent forms of leukemia and lymphoma, i.e. non-Hodgkin's disease, chronic and acute myeloid leukemia (CML / AML), acute lymphocytic leukemia (ALL), Hodgkin's disease, multiple myeloma and T-cell lymphoma. Also included are myelodysplastic syndrome, plasmacytoma, tumor-like syndrome, cancer of unknown primary site, and AIDS-related malignancies.

[0054] In order 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 in conjunction with specific embodiments.

[0055] Embodiment 1:

[0056] Manganese dioxide (MnO 2 ) Formation of Nanoparticles:

[0057] First, tetraethyl orthosilicate (TEOS) was used as raw material to prepare silicon dioxide (SiO 2 ) nanoparticles, 0.75 mL of TEOS was quickly added to a mixed solution containing ethanol (25 mL), ultrapure water (0.5 mL) and ammonia (1.8 mL), and the mixture was stirred at 40° and 150 rpm for 12 h to prepare 80 ± 5 nm SiO 2 Then, the SiO 2 Slowly add 47mL KMnO into the nanoparticle solution. 4 The solution (0.02mmol / L) was reacted under ultrasonic conditions for 6h, and then centrifuged three times (14800rpm, 10min) and the supernatant was discarded to obtain a precipitate. 2 @MnO 2 Nanoparticle precipitation. In order to remove SiO 2 , the prepared SiO 2 @MnO 2 Nanoparticles and Na 2 CO 3The mixture was stirred at 60° and 150 rpm for 12 h, and then the precipitate was washed by centrifugation. The precipitate was dissolved in 40 mL of ultrapure water to obtain 2 mg / mL hollow mesoporous MnO. 2 Hollow mesoporous MnO 2 The particle size and pore size of the ball are Figure 1 As shown, from Figure 1 It can be seen that the hollow mesoporous MnO 2 The average hydrodynamic diameter of the spheres is about 190.1 nm. The desorption branch curves were analyzed by the BJH method to determine the MnO 2 The pore size distribution of the nanoparticles is concentrated at approximately 3.9 nm.

[0058] Example 2

[0059] Intermediate MnO 2 -Vc preparation:

[0060] The prepared MnO 2 Nanoparticles (0.6 mg) were added to 5 mL of ultrapure water containing 10 mg of Vc, and then the mixture was sonicated for 5 min and stirred in the dark for 2 h to obtain MnO 2 -Vc solution.

[0061] Example 3

[0062] Nano drug MnO 2 Preparation of -Vc@mPEG-Ce6:

[0063] In order to improve the water solubility and physiological stability of Ce6, it was modified with PEG to obtain mPEG-Ce6.

[0064] 60 mg Ce6 dissolved in 10 mL DMSO was stirred with 19 mg EDAC (1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride) and 12 mg NHS (N-hydroxysuccinimide) and 200 mg mPEG for 24 h, and the resulting solution was lyophilized. The lyophilized solid was mixed with chloroform to remove unreacted Ce6, and then the precipitate was removed by filtration. Finally, the solvent was removed by a rotary evaporator to obtain mPEG-Ce6. The relative molecular mass of mPEG is 2000.

[0065] Take 1 mL of the prepared mPEG-Ce6 solution and 5 mL of MnO 2 -Vc solution, ultrasonicated for 5 minutes, stirred in the dark for 24 hours, and then centrifuged to remove the supernatant. 2 -Vc@mPEG-Ce6, suspend the finished product in 10 mL of ultrapure water and store at 4 °C.

[0066] Characterization of Nanomedicine MnO Using Transmission Electron Microscopy (TEM) 2 -Vc@mPEG-Ce6 morphology, such as Figure 2 As shown, the prepared nanoparticles have spherical morphology, porous structure and good dispersibility.

[0067] Example 4

[0068] Measurement of Nanomedicine MnO Using a Portable Dissolved Oxygen Meter 2 -Vc@mPEG-Ce6 and H 2 O 2 O released under different conditions 2 Content to verify MnO 2 -Vc@mPEG-Ce6 can effectively alleviate tumor hypoxia. Figure 3 As shown in A, in H 2 O 2 Add MnO to the solution 2 -Vc@mPEG-Ce6 produced O 2 , and dissolved O 2 The amount of MnO 2 -Vc@mPEG-Ce6 concentration increases, indicating that MnO 2 -Vc@mPEG-Ce6 has a catalyst-like activity and can catalyze H 2 O 2 Decomposition produces O 2 At the same time, in another set of experiments, MnO 2 The concentration of -Vc@mPEG-Ce6 was fixed at 200 ppm, and the pH value of the solution was changed. Figure 3 B shows that under acidic conditions, MnO 2 -Vc@mPEG-Ce6 catalyzes H 2 O 2 Produce O 2 In the acidic and hypoxic environment of the tumor, adding MnO 2 -Vc@mPEG-Ce6 can significantly increase the 2 Yield.

[0069] Example 5

[0070] 5,5-Dimethyl-1-pyrroline-N-oxide (DMPO) was used as the capture reagent in 25 mM NaHCO 3 / CO 2 Electron spin resonance (ESR) was performed in buffer to verify the Mn 2+ The ·OH generation capacity. Figure 4As shown in A, a typical ·OH signal (1:2:2:1) was found. Due to the ability of GSH to remove ROS, Mn 2+ +H 2 O 2 The peak of +GSH group is weakened. 2,2,6,6-tetramethyl-4-piperidone (TEMP) is used as the trapping agent and ESR is used for detection 1 O 2 The production of. Figure 4 As shown in B, in all cases exposed to ultrasound (1W / cm 2 , 10 minutes) group were observed to have typical 1 O 2 signal (1:1:1), and at H 2 O 2 The signal was enhanced after treatment. The results showed that the nano drug MnO 2 -Vc@mPEG-Ce6 has a higher ROS production.

[0071] Example 5

[0072] Detection of MnO using 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) 2 -Vc@mPEG-Ce6 has the ability to consume GSH. 2 -Vc@mPEG-Ce6 (0, 50, 100, 200, 300, 400 and 500 μg / mL) were co-cultured with GSH for 30 min, and the MnO 2 -Vc@mPEG-Ce6 was used to deplete GSH (1 mM), and then DTNB (500 μM) was added, and the UV-visible spectra were recorded at wavelengths of 300 to 550 nm. Figure 5 As shown in the figure, the absorption peak intensity at 412 nm increases with the 2 -Vc@mPEG-Ce6 concentration decreased. The results show that MnO 2 -Vc@mPEG-Ce6 can act as a GSH depleting agent to reduce the MnO in cancer cells 2 Completely degraded by GSH.

[0073] Example 6

[0074] Fluorescence microscopy was used to observe the effect of PC-3 cells on MnO 2 -Vc@mPEG-Ce6 uptake. 2 -Vc@mPEG-Ce6 was co-incubated with the fluorescent agent FITC overnight to make MnO 2 -Vc@mPEG-Ce6 was connected with the fluorescent agent FITC, and then the MnO 2-Vc@mPEG-Ce6 was added to the culture medium, and the fluorescence signal of FITC was observed under a fluorescence microscope to evaluate the entry of the nanomedicine into the cells after 0, 1, 2, and 4 hours. Figure 6 As shown, PC-3 cells can effectively take up the nanomedicine MnO 2 -Vc@mPEG-Ce6 and has time dependence.

[0075] Example 7

[0076] CAM / PI double staining method for detecting nano-drug MnO 2 -Vc@mPEG-Ce6 toxicity to PC-3 cells. 2 -Vc@mPEG-Ce6 in vitro antitumor effect, the survival and death of PC-3 cells were detected using CAM / PI double staining kit. PC-3 cells were seeded into six-well plates and allowed to adhere to the wall, and then cultured in a 37°C incubator for 24 hours. The cells were then divided into five treatment groups: 1) control group; 2) MnO 2 (200ppm) group; 3) Ce6 (200ppm) + ultrasound (1w / cm 2 ,3min) group; 4)MnO 2 -Vc@mPEG-Ce6 (200ppm) group; 5) MnO 2 -Vc@mPEG-Ce6(200ppm)+ultrasound(1w / cm 2 After the different treatments, the cells were stained with CAM / PI for 15 minutes according to the manufacturer's instructions. Finally, the fluorescence signal of each group of cells was observed under a fluorescence microscope to determine the cell viability. Figure 7 As shown, nano drug MnO 2 -Vc@mPEG-Ce6 has the strongest killing ability on PC-3 cells under ultrasound.

[0077] Example 8

[0078] First, a BALB / c mouse PC-3 prostate cancer subcutaneous tumor model was established. C-3 cells (1×10 6 ) was suspended in 0.2 mL of saline and injected subcutaneously in the left abdomen of mice to establish a tumor model. 3 At the same time, mice were randomly assigned to different treatment groups for in vivo experiments. First, Cy5.5-loaded MnO 2 -Vc@mPEG-Ce6 nanoparticles to evaluate the 2 -Vc@mPEG-Ce6 biodistribution in vivo. 50 μL of Cy5.5-labeled MnO was injected into the tail vein of mice. 2-Vc@mPEG-Ce6 solution, collect the fluorescence images of mice within 24 hours after injection. BALB / c mice were dissected 24 hours after injection, and the fluorescence images of organs and tumors were collected. Figure 8 As shown in A, the fluorescence intensity of the tumor gradually increased over time, completely accumulated in the tumor site 12 hours after injection, and then decreased due to metabolism in the body (due to the high fluorescence intensity at 0.5 hours, a separate color scale is set). This indicates that MnO 2 The distribution of -Vc@mPEG-Ce6 was time-dependent, which may be due to the enhanced permeability and retention effect of the tumor. Tumors and major organs were also excised 24 hours after injection for ex vivo imaging. Figure 8 B shows that MnO 2 The tumors of mice treated with MnO2-Vc@mPEG-Ce6 showed the strongest Cy5.5 fluorescence signal, while other excised organs no longer had fluorescence, indicating that MnO2-Vc@mPEG-Ce6 nanoparticles were still enriched in the tumor site 24 h after injection.

[0079] Example 9

[0080] When the tumor size reaches about 100 mm 3 Afterwards, PC-3 tumor-bearing mice were randomly assigned to five treatment groups (n=5) and injected with 100 μL of therapeutic drugs via the tail vein, divided into 1) control group; 2) MnO 2 (200ppm) group; 3) Ce6 (200ppm) + ultrasound (1w / cm 2 ,3min) group; 4)MnO 2 -Vc@mPEG-Ce6 (200ppm) group; 5) MnO 2 -Vc@mPEG-Ce6(200ppm)+ultrasound(1w / cm 2 , 3min) group (ultrasound intervention time was 24 hours after injection). The concentration of the drug solution was set at 200ppm. The body weight and tumor volume of mice were recorded within 21 days after different treatments. Fig. 9 A shows that the tumors in the control group grew rapidly, while those in the Ce6+ultrasound group and the MnO 2 -Vc@mPEG-Ce6+ultrasound group had a certain degree of inhibition of tumor growth, among which MnO 2 -Vc@mPEG-Ce6+ultrasound group had the best tumor inhibition effect. At the end of the experiment, the weight of the mice was relatively stable without significant changes, and no statistical difference was found between the groups ( Fig. 9 B). This indicates that MnO 2-Vc@mPEG-Ce6 is biologically safe and has no obvious toxic effects on animals. Tumor images of different treatment groups on day 21 further confirmed the effectiveness of the synergistic treatment of this nanomedicine.

[0081] Example 10

[0082] To further evaluate the effects of different treatments on tumor tissue, GPX4 antibody was used to stain the sections of tumor tissue removed from each group. Fig.10 As shown, compared with the other treatment groups, MnO 2 -Vc@mPEG-Ce6+ultrasound group had the least positive cells (brown), indicating that GPX4 protein expression was downregulated, which proved that the nanodrug MnO 2 -Vc@mPEG-Ce6 causes ferroptosis of tumor cells through the GPX4 pathway.

[0083] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A sono / chemical synergistic anti-tumor nanomedicine, characterized in that: The nanomedicine uses hollow mesoporous MnO2 nanoparticles as carriers and loads vitamin C and sonosensitizer dihydrochlorin e6.

2. The sono / chemodynamic synergistic anti-tumor nanomedicine according to claim 1, characterized in that: The MnO2 nanoparticles are MnO2 nanospheres; Preferably, the particle size of the MnO2 nanospheres is about 190 nm; and the pore size is about 3.9 nm.

3. The method for preparing the sono / chemodynamic synergistic anti-tumor nanomedicine according to any one of claims 1 to 2, characterized in that: include: (1) preparing silica nanoparticles using tetraethyl orthosilicate as a raw material, then adding KMnO4 solution to the silica nanoparticle solution to obtain SiO2@MnO2 nanoparticles, and using the SiO2@MnO2 nanoparticles as a template to remove SiO2 to obtain hollow mesoporous MnO2 nanospheres; (2) dispersing the hollow mesoporous MnO2 nanospheres in a vitamin C solution, and reacting in a dark and stirring manner to obtain a nanodrug intermediate MnO2-Vc; (3) The sonosensitizer dihydrochlorin e6 was modified with polyethylene glycol to obtain mPEG-Ce6, and mPEG-Ce6 was mixed with the nanodrug intermediate MnO2-Vc. The mixture was stirred in the dark to obtain the nanodrug MnO2-Vc@mPEG-Ce6.

4. The preparation method according to claim 3, characterized in that: In step (1), the molar ratio of silica nanoparticles to KMnO4 is 2.5 to 3:1, preferably 2.85:1; Or, in step (1), the concentration of the KMnO4 solution is 0.015 to 0.03 mmol / L, preferably 0.02 mmol / L; Alternatively, in step (1), the ultrasonic power for obtaining SiO2@MnO2 nanoparticles is 140-160W, preferably 150W; and the time is 5-7h, preferably 6h.

5. The preparation method according to claim 3, characterized in that: In step (1), Na2CO3 is used to etch and remove SiO2, and the concentration of Na2CO3 is 0.015-0.03mmol / L, preferably 0.02mmol / L; the etching temperature is 55-65°C, preferably 60°C; the etching time is 10-15h, preferably 12h; the stirring rate during etching is 120-200rpm, preferably 150rpm.

6. The preparation method according to claim 3, characterized in that: In step (2), the mass ratio of hollow mesoporous MnO2 nanospheres to vitamin C is 0.03-0.01:1, preferably 0.05:1; Or, in step (2), the concentration of vitamin C in the vitamin C solution is 1 to 3 g / L, preferably 2 g / L; Alternatively, in step (2), the reaction temperature is room temperature while stirring in the dark, and the stirring time is 10 to 15 hours, preferably 12 hours.

7. The preparation method according to claim 3, characterized in that: In step (3), the mass ratio of Ce6 to MnO2 is 8 to 12:1, preferably 10:1; Alternatively, in step (3), the reaction temperature is room temperature while stirring in the dark, and the stirring time is 20 to 30 hours, preferably 24 hours.

8. Use of the sono / chemodynamic synergistic anti-tumor nanomedicine according to any one of claims 1 to 2 and / or the sono / chemodynamic synergistic anti-tumor nanomedicine prepared by the preparation method according to any one of claims 3 to 7 in the preparation of anti-tumor drugs.

9. An anti-tumor drug composition, characterized in that: The invention comprises the sono / chemical synergistic anti-tumor nanomedicine as described in any one of claims 1 to 2 and / or the sono / chemical synergistic anti-tumor nanomedicine prepared by the preparation method as described in any one of claims 3 to 7.

10. The antitumor pharmaceutical composition according to claim 9, characterized in that: The anti-tumor drug composition also includes drugs related to tumor treatment; Preferably, the tumor treatment-related drug is an anti-tumor drug, an immune adjuvant, a checkpoint inhibitor or an antigen protein.

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