Acidic and glutathione responsive manganese-based nanomaterial and preparation method and application thereof

Abstract

The present invention discloses an acidic and glutathione-responsive manganese-based nanomaterial and its preparation method and application. The preparation method of the nanoparticles is as follows: 1. Adding photosensitizer dihydrochlorin to dimethyl sulfoxide, adding to methanol after complete dissolution, then adding manganese acetylacetonate and zinc acetate, mixing evenly, and obtaining liquid A; 2. Adding 2-methylimidazole and 2-(trifluoromethyl)-1H-imidazole to methanol, mixing evenly, then adjusting the pH to 9-10, and obtaining liquid B; 3. Adding liquid B to liquid A under stirring, then continuing to stir for 1-3 hours, standing, centrifuging, and drying to obtain Ce6@Mn-F-ZIF-8; 4. Dispersing nanoparticles Ce6@Mn-F-ZIF-8 in PBS, adding 4T1 cell membrane protein, and squeezing with a nano extruder after ultrasonication for 30-60 minutes to obtain Ce6@Mn-F-ZIF-8@Mem. The nanoparticles can be specifically activated through a series of enzyme-linked reactions in the tumor microenvironment to achieve the target of tumor area. 1 H / 19 F MRI and in vivo fluorescence imaging can also catalyze the production of oxygen to improve the hypoxic environment of the tumor, and produce ROS to kill tumor cells.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedical imaging, and in particular relates to an acidic and glutathione responsive manganese-based nanomaterial and a preparation method and application thereof. Background Art

[0002] The incidence of breast cancer ranks first among female malignant tumors. The symptoms of breast cancer are mostly not obvious. In advanced breast cancer, cancer cells may metastasize to the original site and cause multi-organ lesions throughout the body, directly threatening the patient's life. Among the many subtypes of breast cancer, triple-negative breast cancer has the characteristics of poor prognosis, poor invasiveness, high metastasis and high mortality, and is insensitive to commonly used endocrine therapy and targeted therapy. Triple-negative breast cancer also has its own significant characteristics, such as high tumor mutation load, high PD-L1 expression and high proportion of tumor-infiltrating lymphocytes, so immunotherapy may be effective for it.

[0003] In recent years, tumor immunotherapy has been shown to be a promising treatment method that can induce the body's innate immune system to kill tumor cells. However, to date, most immunotherapies have limited effects, patient response rates remain low (<20%), individual responses to treatment vary greatly, and systemic immunity is often activated, leading to severe immunotoxicity such as cytokine release syndrome. At present, nanocatalyst-based tumor therapy has made some progress, but achieving precise treatment targeting the tumor-specific microenvironment (TME) remains a major challenge.

[0004] At present, imaging examinations for breast cancer are mainly performed through breast mammography, breast ultrasound, and breast magnetic resonance imaging. When the imaging of suspected breast cancer is unclear, a tissue biopsy is performed to make a final diagnosis. Tissue biopsy is invasive for patients, and current imaging examinations have the risk of false positives and lesions that are too small to be identified by imaging. Summary of the invention

[0005] In order to solve the problems existing in the above-mentioned prior art, the present invention provides an acidic and glutathione-responsive manganese-based nanomaterial and its preparation method and application, wherein the manganese-based nanomaterial can remain inert (closed) in normal tissues, and only in the tumor microenvironment can it be specifically activated (opened) through a series of enzyme-linked reactions to achieve tumor microenvironmental regulation. 1 H / 19While performing F MRI and in vivo fluorescence imaging, it can also catalyze the production of oxygen to improve the hypoxic environment of the tumor, and produce ROS to kill tumor cells. More importantly, the manganese-based nanomaterial enhances oxidative stress in the tumor area, reverses the immunosuppressive TME, and promotes anti-tumor immune response. At the same time, the photodynamic / chemodynamic synergistic therapy initiated by the manganese-based nanomaterial combined with PD-L1 antibody can effectively inhibit the growth of primary tumors and tumor metastasis.

[0006] The technical solution adopted to achieve the above-mentioned purpose of the present invention is:

[0007] A method for preparing an acidic and glutathione-responsive manganese-based nanomaterial comprises the following steps:

[0008] S 1 Preparation of Ce6@Mn-F-ZIF-8:

[0009] S 1.1 , adding the photosensitizer chlorin to dimethyl sulfoxide, adding to methanol after complete dissolution to obtain a chlorin solution, adding manganese acetylacetonate and zinc acetate to the chlorin solution, the molar ratio of chlorin, manganese salt and zinc acetate being 1:8:7-1:9:8, mixing evenly to obtain a solution A;

[0010] S 1.2 , adding 2-methylimidazole and 2-(trifluoromethyl)-1H-imidazole to methanol, wherein the molar ratio of zinc acetate, 2-methylimidazole and 2-(trifluoromethyl)-1H-imidazole is 1:10:2-1:11:3, mixing well, and then adjusting the pH to 9-10 to obtain solution B;

[0011] S 1.3 , add liquid B into liquid A under stirring, then continue stirring for 1-3 hours, let stand for about 24 hours, centrifuge after standing, dry the obtained solid, and obtain the nanomaterial Ce6@Mn-F-ZIF-8;

[0012] S 2 Preparation of Ce6@Mn-F-ZIF-8@Mem:

[0013] The nanomaterial Ce6@Mn-F-ZIF-8 was dispersed in PBS, and 4T1 cell membrane protein was added. The mass ratio of Ce6@Mn-F-ZIF-8 to 4T1 cell membrane protein was 3:1-6:1, and ultrasonic treatment was performed for 30-60 minutes. After the ultrasonic treatment, the nano-extruder was used to extrude the nanomaterial Ce6@Mn-F-ZIF-8@Mem, which was acidic and glutathione responsive.

[0014] Further, step S 1.2 In the solution, the pH was adjusted to 9-10 with aqueous ammonia.

[0015] Furthermore, the manganese-based nanomaterial has Mn-F-ZIF-8 loaded with photosensitizer dihydrochlorin as the core and 4T1 cell membrane protein as the shell.

[0016] Application of an acidic and glutathione-responsive manganese-based nanomaterial in the preparation of multimodal imaging contrast agents and chemodynamic and photodynamic therapy drugs.

[0017] Furthermore, the multimodal imaging includes 1 H imaging, 19 F MRI imaging and in vivo fluorescence imaging.

[0018] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0019] 1. The present invention introduces Mn, 4-(trifluoromethyl)-1H-imidazole and photosensitizer dihydrochlorin Ce6 into the ZIF-8 system through a "one-pot method" to obtain Ce6@Mn-F-ZIF-8 nanoparticles, and modifies the tumor cell membrane on the surface of Ce6@Mn-F-ZIF-8 by ultrasound and repeated extrusion to obtain the final material Ce6@Mn-F-ZIF-8@Mem. It can be seen that the preparation method is simple, easy to operate, and has little pollution to the environment.

[0020] 2. If Figure 7 As shown in the figure, the nanomaterial can efficiently accumulate in the tumor area, and due to the characteristics of the tumor microenvironment - slightly acidic and high glutathione, Ce6@Mn-F-ZIF-8@Mem is cleaved. After cleavage, the following changes occur: 1. The Mn bound in the ZIF metal structure is released into the environment, and glutathione converts Mn into 3+ , Mn 4+ Reaction to Mn 2+ ,accomplish 1 The transition of HMRI from "off" to "on"; 2, 4-(trifluoromethyl)-1H-imidazole and Mn 2+ The distance between them changes, Mn 2+ The effect on 4-(trifluoromethyl)-1H-imidazole is weakened. 19 F MRI signal changes from "off" to "on"; 3. During the cleavage process, Ce6 encapsulated in the nanoparticles is released, which can be used for in vivo fluorescence imaging, and can also be used to irradiate the tumor area with 660nm laser to react with Mn 2+ The Fenton reaction works synergistically to promote the transformation of M2 macrophages to M1 macrophages, thereby achieving immune activation in the tumor area.

[0021] 3. The nanomaterial is designed to achieve tumor microenvironment (slightly acidic, high GSH) response and achieve 1 H MRI,19 FMRI and multimodal responsive imaging of in vivo fluorescence from "off" to "on" can combine the advantages of three imaging modes: 1 H MRI has high spatial resolution and high tissue contrast; 19 F MRI has no interference from background signals and is highly selective; in vivo fluorescence imaging has high sensitivity and short sampling time. On the other hand, the nanoparticles can realize responsive imaging of the tumor area from "off" to "on", can realize specific response lighting to the tumor location, and make the lesion area clearer, reduce interference from non-lesion areas, and achieve more accurate diagnosis. In addition, multi-modal imaging can be performed, and multiple imaging complements each other, thereby obtaining multi-angle and multi-scale imaging information, accurately locating the tumor area, and improving the accuracy of lesion diagnosis.

[0022] 4. The tumor cell membrane modified on the surface of the nanomaterial gives it the ability to escape immunity and has a longer blood circulation time. In addition, due to the breast cancer tumor cell membrane modified on the surface of the material, based on the homologous targeting effect, the material is more conducive to enrichment in the breast cancer area and has high biosafety.

[0023] 5. The nanomaterial enhances the oxidative stress of tumors, reverses the immunosuppressive TME, and promotes anti-tumor immune response through combined treatment of chemokinetics (CDT) and photodynamic therapy (PDT). CDT uses the slightly acidic and high redox properties of the tumor microenvironment to produce ROS in the Fenton reaction to kill tumors. CDT is highly selective for tumors, non-toxic to normal tissues, has no drug resistance, and is not limited by tissue depth, but its therapeutic effect is affected by the Fenton reaction rate. PDT relies on the uptake of photosensitizers by the tumor site. The photosensitizers are excited and transfer energy to O in the tissue. 2 , converting it into highly toxic substances ROS to treat tumors. The advantage of PDT is that it can achieve selective treatment of tumor sites by controlling the irradiation of light sources, but PDT is limited by the light source and is currently mostly used to treat tumors on the surface of the body. Experiments have shown that combining CDT with PDT to treat breast cancer can achieve complementary advantages and improve the treatment effect of breast cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 TEM image of Ce6@Mn-F-ZIF-8@Mem prepared in Example 1.

[0025] Figure 2 Zeta potential diagram of Ce6@Mn-F-ZIF-8 and Ce6@Mn-F-ZIF-8@Mem prepared in Example 1.

[0026] Figure 3The living bodies of Ce6@Mn-F-ZIF-8@Mem prepared in Example 1 at different times 1 HMRI imaging.

[0027] Figure 4 The living body of Ce6@Mn-F-ZIF-8@Mem prepared in Example 1 19 F MRI imaging.

[0028] Figure 5 These are the in vivo fluorescence imaging images of Ce6@Mn-F-ZIF-8@Mem prepared in Example 1 at different times.

[0029] Figure 6 The CDT and PDT treatment effect diagram of Ce6@Mn-F-ZIF-8@Mem prepared in Example 1, wherein 1 represents the control group; 2 represents the first test group; and 3 represents the second test group.

[0030] Figure 7 This is the application principle diagram of Ce6@Mn-F-ZIF-8@Mem prepared in Example 1.

[0031] Figure 8 This is the TEM image of Ce6@Mn-F-ZIF-8 prepared in Comparative Example 1.

[0032] Fig. 9 This is the TEM image of Ce6@Mn-F-ZIF-8 prepared in Comparative Example 2. DETAILED DESCRIPTION

[0033] The present invention is described in detail below with reference to specific embodiments.

[0034] Example 1

[0035] S 1 Preparation of Ce6@Mn-F-ZIF-8:

[0036] S 1.1 1. Add 50 mg of photosensitizer chlorin Ce6 (0.084 mmol) into 0.1 mL of dimethyl sulfoxide, and after complete dissolution, add into 50 mL of methanol to obtain chlorin solution. Weigh 176.13 mg of manganese acetylacetonate (0.70 mmol) and 110 mg (0.60 mmol) of zinc acetate and add into the chlorin solution. After ultrasonic dispersion, stir evenly to obtain solution A.

[0037] S 1.2 525.44 mg 2-methylimidazole (6.4 mmol) and 217.728 mg 2-(trifluoromethyl)-1H-imidazole (1.6 mmol) were weighed and added to 50 mL methanol, ultrasonically dispersed evenly, and ammonia water was added dropwise to adjust the pH to 10 to obtain solution B;

[0038] S 1.3 , add liquid B into liquid A under stirring, then continue stirring for 1 hour, let stand for 24 hours, centrifuge, and dry the obtained solid to obtain nanoparticles Ce6@Mn-F-ZIF-8;

[0039] S 2 , Extraction of 4T1 cell membrane proteins (using Solarbio EX1110 membrane protein extraction kit for extraction)

[0040] S 2.1 , take 6×10 6 Mouse breast cancer cells were centrifuged at 4°C and 500×g for 5 minutes, the culture medium was carefully aspirated, and the cells were collected as dry as possible. The cells were washed twice with cold PBS, and the supernatant was aspirated as dry as possible after each wash to obtain a cell sample.

[0041] S 2.2 , add 300 μL of cold reagent A to the cell sample, mix well, shake at 5°C for 25 minutes until the cells are fully lysed and the cell precipitate is significantly reduced, then centrifuge at 4°C and 12,000×g for 5 minutes, transfer the supernatant to another clean centrifuge tube, incubate at 37°C in a water bath for 8 minutes, and then incubate at 37°C and 1,000×g for 30 minutes.

[0042] Centrifuge at × g for 5 minutes, at which time the solution is divided into two layers, the lower layer is membrane protein, about 40 μL, carefully remove the upper liquid, and collect the upper part for standby analysis;

[0043] S 2.3 Use 200 μL of ice-cold reagent B to fully dissolve the lower membrane protein. After mixing, place it in an ice bath for 2 minutes, then in a 37°C water bath for 8 minutes, and then centrifuge it at 37°C and 1000×g for 5 minutes. At this time, the solution is divided into two layers. The lower layer is membrane protein, about 40 μL. Carefully remove the upper layer liquid and retain the lower membrane protein.

[0044] S 2.4 Extract the lower membrane protein again according to the above method and steps (this step is optional and generally not necessary).

[0045] S 2.5 The lower membrane protein was dissolved with 100 μL of cold reagent C to obtain 4T1 cell membrane protein. The 4T1 cell membrane protein was quantified and packaged in a -80°C refrigerator, and freeze-dried in a freeze dryer for downstream experiments.

[0046] S 3 Preparation of Ce6@Mn-F-ZIF-8@Mem:

[0047] 5 mg of nanoparticles Ce6@Mn-F-ZIF-8 were dispersed in 2 mL of PBS, 1 mg of 4T1 cell membrane protein was added, and ultrasonic treatment was performed for 30 minutes. After the ultrasonic treatment, the nanoparticles were repeatedly extruded using a nano-extruder to obtain the acidic and glutathione-responsive manganese-based nanomaterial Ce6@Mn-F-ZIF-8@Mem. The manganese-based nanomaterial has Mn-F-ZIF-8 loaded with photosensitizer dihydrochlorin as the core and 4T1 cell membrane protein as the shell.

[0048] The Ce6@Mn-F-ZIF-8@Mem prepared in this example was scanned by a transmission electron microscope, and the obtained TEM image is as follows: Figure 1 As shown by Figure 1 It can be seen that the Ce6@Mn-F-ZIF-8@Mem nanoparticles prepared in this embodiment have a uniform particle size of about 300 nm and an irregular polyhedral morphology. Figure 1 It can be seen that the surface of the nanoparticle material is rough and covered with membrane proteins, indicating that the 4T1 cell membrane proteins have been successfully modified.

[0049] The Ce6@Mn-F-ZIF-8 (abbreviated as MF8) and Ce6@Mn-F-ZIF-8@Mem (abbreviated as MF8T) prepared in this example were tested for Zeta potential. The Zeta potentials obtained are shown in Figure 2 As shown by Figure 2 It can be seen that before modifying the 4T1 cell membrane protein, the potential of Ce6@Mn-F-ZIF-8 was positive, and after modifying the 4T1 cell membrane, the potential of Ce6@Mn-F-ZIF-8@Mem was negative, indicating that the 4T1 cell membrane was successfully modified on the surface of Ce6@Mn-F-ZIF-8.

[0050] Experiment 1: Living Ce6@Mn-F-ZIF-8@Mem Nanoparticles of the Present Invention 1 H MRI test test method:

[0051] The Ce6@Mn-F-ZIF-8@Mem prepared in Example 1 was dispersed in sterile PBS to prepare a contrast agent with a concentration of 500 ug / mL. The contrast agent was then imaged in vivo on tumor-bearing mice using a 9.4T small animal magnetic resonance imaging system. 1 HMRI imaging, and then the contrast agent was injected into the tumor-bearing mice through the tail vein. After injection, the tumor-bearing mice were examined in vivo using a 9.4T small animal magnetic resonance imaging instrument at 2h, 4h, 8h, 12h, 24h, and 36h. 1 H MRI imaging.

[0052] Test results:

[0053] After the contrast agent was injected into the tail vein, no death or other toxic side effects occurred in the mice, indicating that the Ce6@Mn-F-ZIF-8@Mem of the present invention has good biosafety.

[0054] Live images of tumor-bearing mice at different time points after contrast agent injection 1 HMRI imaging Figure 3 As shown, from Figure 3 It can be seen that the brightness of the tumor area of ​​the tumor-bearing mice changes from weak to strong and then to weak again, indicating that after the contrast agent is injected into the tail vein, it is enriched in the tumor area of ​​the tumor-bearing mice through the blood circulation due to the EPR effect and the active targeting effect of the surface-modified membrane protein. In addition, the analysis of the MRI signal intensity of the tumor area shows that the tumor area is brightest at 8h, indicating that the tumor cell membrane modified on the surface of the nanoparticles gives it the ability to escape immunity and can achieve long circulation in the mouse body.

[0055] Experiment 2: Living Ce6@Mn-F-ZIF-8@Mem Nanoparticles of the Present Invention 19 F MRI test test method:

[0056] The Ce6@Mn-F-ZIF-8@Mem prepared in Example 1 was dispersed in sterile PBS to prepare a contrast agent with a concentration of 500 ug / mL. The contrast agent was injected into the tumor of the tumor-bearing mice by intratumoral injection. After injection, the tumor of the tumor-bearing mice was imaged in vivo using a 9.4T small animal magnetic resonance imaging device. 19 F MRI imaging and 1 H MRI imaging.

[0057] Test results:

[0058] After in situ injection of the drug solution, the health status of the tumor-bearing mice was not affected, indicating that the Ce6@Mn-F-ZIF-8@Mem of the present invention has high biological safety.

[0059] After contrast agent was injected into the tumor area, Ce6@Mn-F-ZIF-8@Mem disintegrated in the tumor area due to the slightly acidic and high GSH environment of the tumor area. 2+ The distance between Mn and the fluorinated small molecule 4-(trifluoromethyl)-1H-imidazole changes. 2+ Yes -CF 3 The effect of group relaxation is weakened, so 19 F MRI signal is evident in the tumor region.

[0060] After contrast agent injection, the living tissue of tumor-bearing mice 19 F MRI imaging and 1 H MRI imaging Figure 4 As shown, from Figure 4 It can be seen that after contrast agent was injected into the tumor site of tumor-bearing mice, there was a significant and bright 19 F signal, while other regions have no response conditions of Ce6@Mn-F-ZIF-8@Mem. 19 F signal. This shows that Ce6@Mn-F-ZIF-8@Mem of the present invention has 19 F MRI is specific to tumor sites and can eliminate false positives in tumor diagnosis, thus achieving more accurate early diagnosis of tumors.

[0061] Experiment 3: In vivo fluorescence imaging test of Ce6@Mn-F-ZIF-8@Mem nanoparticles of the present invention Experimental method:

[0062] The Ce6@Mn-F-ZIF-8@Mem prepared in Example 1 was dispersed in sterile PBS to prepare a contrast agent with a concentration of 500 ug / mL. The tumor-bearing mice were imaged with live fluorescence using a small animal in vivo fluorescence imager. The contrast agent was then injected into the tumor-bearing mice via the tail vein. After injection, the tumor-bearing mice were imaged with live fluorescence using a small animal in vivo fluorescence imager at 0.5 h, 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, and 48 h.

[0063] Test results:

[0064] In vivo fluorescence imaging of tumor-bearing mice at different time points after contrast agent injection Figure 5 As shown, from Figure 5 It can be seen that at this color bar intensity, there is no in vivo fluorescence signal except in the tumor area. On the one hand, it shows that the nanoparticles can efficiently target the tumor area and stay in the tumor area; on the other hand, it shows that the nanoparticles release Ce6 in response to the slightly acidic and high GSH environment in the tumor area, and other areas do not meet the conditions for the disintegration of the nanoparticles.

[0065] In addition, from Figure 5 It can also be seen that the fluorescence imaging of the tumor area experienced a process from weak to strong and then to weak again. The fluorescence intensity of the tumor area reached its maximum value at 8 hours, and there was a long window observation period, which facilitated the diagnosis and treatment of tumor mice.

[0066] Experiment 4: Ce6@Mn-F-ZIF-8@Mem nanoparticles activation immune test of the present invention

[0067] Test method:

[0068] The Ce6@Mn-F-ZIF-8@Mem prepared in Example 1 was dispersed in sterile PBS and prepared into a working solution with a concentration of 500ug / mL. The tumor-bearing mice were randomly divided into a control group, a test group 1 and a test group 2, with 3 tumor-bearing mice in each group. The control group was injected with sterile PBS, the test group 1 was injected with the working solution, and the test group 2 was injected with the working solution and irradiated with a 660nm laser at the same time. Each group was injected once every other day, and the treatment time was 7 days. After the treatment, the tumors of each group of tumor-bearing mice were dissected out, and the content of M1 macrophages and M2 macrophages in the tumor was detected by flow cytometry after treatment.

[0069] Test results:

[0070] The CDT and PDT treatment effects of Ce6@Mn-F-ZIF-8@Mem prepared in Example 1 are as follows Figure 6 As shown by Figure 6 It can be seen that Ce6@Mn-F-ZIF-8@Mem can produce toxic ROS through Fenton reaction to achieve chemodynamic therapy; Ce6 as a photosensitizer is excited under specific laser irradiation and transfers energy to O in the tissue. 2 , converting it into ROS to treat tumors and achieve photodynamic therapy. Ce6@Mn-F-ZIF-8@Mem+Laser can combine CDT with PDT to achieve the effect of combined treatment. Therefore, the combined treatment group has the best therapeutic effect, achieving the polarization of M2 cells into M1 cells and activating the immune response in the tumor area.

[0071] Comparative Example 1

[0072] S 1 Preparation of Ce6@Mn-F-ZIF-8:

[0073] S 1.1 1. Add 50 mg of photosensitizer chlorin Ce6 (0.084 mmol) into 0.1 mL of dimethyl sulfoxide, and after complete dissolution, add into 20 mL of methanol to obtain chlorin solution. Weigh 89.475 mg (0.50 mmol) of manganese nitrate and 594.98 mg (2 mmol) of zinc nitrate hexahydrate and add into the chlorin solution. After ultrasonic dispersion, stir evenly to obtain solution A.

[0074] S 1.2 3119.8 mg 2-methylimidazole (38 mmol) and 217.728 mg 2-(trifluoromethyl)-1H-imidazole (1.6 mmol) were weighed and added to 80 mL methanol, and ultrasonically dispersed uniformly. Ammonia water was added dropwise to adjust the pH to 10 to obtain solution B;

[0075] S 1.3, add liquid B into liquid A under stirring, then continue stirring for 1 hour, let stand for 24 hours, centrifuge, and dry the obtained solid to obtain nanoparticles Ce6@Mn-F-ZIF-8;

[0076] The Ce6@Mn-F-ZIF-8 prepared in this comparative example was scanned by a transmission electron microscope, and the obtained TEM image is as follows: Figure 8 As shown by Figure 8 It can be seen that the particle size of the Ce6@Mn-F-ZIF-8@Mem nanomaterial prepared in this comparative example is about 20 nm, which is too small and is not conducive to subsequent in vivo experiments and is prone to produce the EPR effect.

[0077] Comparative Example 2

[0078] S 1 Preparation of Ce6@Mn-F-ZIF-8:

[0079] S 1.1 1. Add 50 mg of photosensitizer chlorin Ce6 (0.084 mmol) into 0.1 mL of dimethyl sulfoxide, and after complete dissolution, add into 50 mL of methanol to obtain chlorin solution. Weigh 86.5 mg (0.50 mmol) of manganese acetate and 594.98 mg (2 mmol) of zinc nitrate hexahydrate and add into the chlorin solution. After ultrasonic dispersion, stir evenly to obtain solution A.

[0080] S 1.2 3119.8 mg 2-methylimidazole (38 mmol) and 217.728 mg 2-(trifluoromethyl)-1H-imidazole (1.6 mmol) were weighed and added to 50 mL methanol, and ultrasonically dispersed uniformly. Ammonia water was added dropwise to adjust the pH to 10 to obtain solution B;

[0081] S 1.3 , add liquid B into liquid A under stirring, then continue stirring for 1 hour, let stand for 24 hours, centrifuge, and dry the obtained solid to obtain nanoparticles Ce6@Mn-F-ZIF-8;

[0082] The Ce6@Mn-F-ZIF-8 prepared in this comparative example was scanned by a transmission electron microscope, and the obtained TEM image is as follows: Fig. 9 As shown by Fig. 9 It can be seen that the Ce6@Mn-F-ZIF-8 nanomaterial prepared in this comparative example is adhered, which is not conducive to obtaining the membrane structure by extruding the filtration membrane in the subsequent cell membrane wrapping process.

Claims

1. A method for preparing an acidic and glutathione-responsive manganese-based nanomaterial, characterized in that The steps include: S1. Preparation of Ce6@Mn-F-ZIF-8: S 1.1 , adding the photosensitizer chlorin to dimethyl sulfoxide, adding to methanol after complete dissolution to obtain a chlorin solution, adding manganese acetylacetonate and zinc acetate to the chlorin solution, wherein the molar ratio of chlorin, manganese acetylacetonate and zinc acetate is 1:8:7-1:9:8, and mixing evenly to obtain a solution A; S 1.2 , adding 2-methylimidazole and 2-(trifluoromethyl)-1H-imidazole to methanol, wherein the molar ratio of zinc acetate, 2-methylimidazole and 2-(trifluoromethyl)-1H-imidazole is 1:10:2-1:11:3, mixing well, and then adjusting the pH to 9-10 to obtain solution B; S 1.3 , add liquid B into liquid A under stirring, then continue stirring for 1-3 h, let stand, centrifuge after standing, dry the obtained solid, and obtain the nanomaterial Ce6@Mn-F-ZIF-8; S2. Preparation of Ce6@Mn-F-ZIF-8@Mem: The nanomaterial Ce6@Mn-F-ZIF-8 was dispersed in PBS, and 4T1 cell membrane protein was added, the mass ratio of Ce6@Mn-F-ZIF-8 to 4T1 cell membrane protein was 3:1-6:1, and ultrasonic treatment was performed for 30-60 min. After the ultrasonic treatment, it was extruded with a nano extruder to obtain the acidic and glutathione responsive manganese-based nanomaterial Ce6@Mn-F-ZIF-8@Mem.

2. The method for preparing the acidic and glutathione responsive manganese-based nanomaterial according to claim 1, characterized in that: Step S 1.2 In the solution, the pH was adjusted to 9-10 with aqueous ammonia.

3. An acidic and glutathione-responsive manganese-based nanomaterial prepared by any of the methods of claims 1-2, characterized in that: The manganese-based nanomaterial has Mn-F-ZIF-8 loaded with photosensitizer dihydrochlorin as the core and 4T1 cell membrane protein as the shell.

4. Use of the acidic and glutathione-responsive manganese-based nanomaterial according to claim 3 in the preparation of multimodal imaging contrast agents and chemodynamic and photodynamic therapy drugs.

5. The use of the acidic and glutathione responsive manganese-based nanomaterial according to claim 4, characterized in that: The multimodal imaging includes 1 H imaging, 19 F MRI imaging and in vivo fluorescence imaging.

Images