A functionalized manganese-doped mesoporous nanomaterial and its preparation method and application

Abstract

The present invention relates to a functionalized manganese-doped mesoporous nanomaterial, its preparation method, and application. The preparation method specifically comprises: preparing solid silica nanospheres; mixing the solid silica nanospheres with maleate and manganese salt aqueous solutions, and preparing manganese-doped mesoporous silica hollow spheres using a hydrothermal method; loading isoniazid and benzothiazole sulfinate into the manganese-doped mesoporous silica hollow spheres to obtain a functionalized manganese-doped mesoporous silica nanomaterial; and compounding the cell membranes of treated mouse breast cancer cells with the functionalized manganese-doped mesoporous silica nanomaterial to obtain a cell membrane-modified functionalized manganese-doped mesoporous silica nanomaterial. Compared with the prior art, the preparation method of the present invention is simple, and the prepared composite nanomaterial has magnetic resonance imaging contrast performance and a good synergistic therapeutic effect. It can be used for magnetic resonance imaging-guided tumor treatment, achieving integrated diagnosis and treatment.

Description

Technical Field

[0001] The present invention belongs to the technical field of nanomaterials and molecular imaging, and relates to a functionalized manganese-doped mesoporous nanomaterial and a preparation method and application thereof. Background Art

[0002] Singlet oxygen ( 1 Reactive oxygen species (ROS) such as O2) and hydroxyl radicals (·OH) can induce damage to cell contents (such as proteins, lipids and DNA), leading to cell apoptosis or necrosis. Reactive oxygen species (ROS)-mediated cancer treatment has received widespread attention, such as photodynamic therapy (PDT) and chemodynamic therapy (CDT). PDT uses light and photosensitizers to generate ROS to specifically kill cancer cells. However, the low penetration depth of lasers makes it only effective for superficial tumors and difficult to reach deep tumor sites. CDT is a method that uses some lasers with metal ions (such as Fe 2+ 、Cu 2+ and Mn 2+ ) nanomaterials undergo Fenton or Fenton-like reactions with H2O2 overexpressed in the tumor site to produce ·OH, thereby leading to cancer cell apoptosis.

[0003] Patent CN109010850A discloses a method for preparing and using a curcumin-loaded glucan-modified hollow mesoporous silica nanomaterial doped with gadolinium. The preparation method comprises: 1) preparing uniformly dispersed solid silica nanospheres with uniform particle size; 2) uniformly mixing the aqueous solution of solid silica nanospheres obtained in step 1) with sodium acetate and gadolinium nitrate, and transferring the mixture to a polytetrafluoroethylene-lined reactor to hydrothermally prepare hollow mesoporous silica nanospheres doped with the rare earth metal gadolinium; 3) loading curcumin into the cavities of the hollow mesoporous silica nanospheres obtained in step 2) by stirring, and modifying amino groups on their surfaces by adding 3-aminopropyltriethoxysilane and continuing stirring; and 4) activating carboxyl groups on the surface of the glucan with EDC / NHS, which then binds to the amino groups of the material obtained in step 3) to prepare the curcumin-loaded glucan-modified hollow mesoporous silica nanomaterial. However, this patent has poor targeting effect and cannot be released specifically in tumor cells; moreover, the sonodynamic therapy used requires exogenous ultrasonic stimulation.

[0004] Patent CN113493223A discloses a method for preparing and applying hollow manganese dioxide nanospheres. The method involves first preparing template silica nanospheres. Using the silica nanospheres as templates, SiO2@MnO2 nanospheres with a core-shell structure are hydrothermally synthesized. Hollow manganese dioxide nanospheres are then etched with sodium hydroxide to form hollow manganese dioxide nanospheres. By coating the surface of these nanospheres with tumor cell membranes, targeted tumor cell therapy can be achieved. The synthesis process is simple, and the product has a uniform particle size, high dispersion, good water solubility, and stable properties. However, the patent's Fenton-like reaction has low efficiency and requires a large number of reactants, resulting in insufficient CDT therapeutic efficacy. Therefore, a new, efficient ·OH generation strategy is urgently needed. Summary of the Invention

[0005] The purpose of the present invention is to overcome the defects of the common excitation light sources (visible light and near-infrared light) in the above-mentioned prior art, such as the penetration depth of less than 1 cm in tissues, limited efficacy on solid tumors and deep tumors, low efficiency of Fenton-like reactions and a large number of required reactants, and to provide a functionalized manganese-doped mesoporous nanomaterial and its preparation method and application. The preparation method of the present invention is simple, and the prepared composite nanomaterial has magnetic resonance imaging contrast performance and good synergistic therapeutic effect. It can be used for magnetic resonance imaging-guided tumor treatment to achieve integrated diagnosis and treatment.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] One of the technical solutions of the present invention is to provide a method for preparing a functionalized manganese-doped mesoporous nanomaterial, the method comprising the following steps:

[0008] (1) Preparation of uniform and monodispersed solid silica nanospheres;

[0009] (2) mixing solid silica nanospheres and aqueous solutions of maleate and manganese salts evenly, and preparing manganese-doped mesoporous silica hollow spheres by a hydrothermal method;

[0010] (3) Isoniazid (INH) and sulfur dioxide prodrug benzothiazole sulfinate (BTS) were loaded into the cavities and pores of manganese-doped mesoporous silica hollow spheres to obtain functionalized manganese-doped mesoporous silica nanomaterials;

[0011] (4) The cell membrane of the treated mouse breast cancer cells (4T1 cells) was evenly compounded with the functionalized manganese-doped mesoporous silica nanomaterial to obtain the cell membrane-modified functionalized manganese-doped mesoporous silica nanomaterial.

[0012] Furthermore, step (1) includes the following steps:

[0013] (1.1) Stir and mix anhydrous ethanol, ammonia (NH3·H2O), and deionized water until uniformly mixed and heat.

[0014] (1.2) When the temperature is stable, tetraethyl orthosilicate (TEOS) is added dropwise to the solution obtained in step (1.1) via a syringe pump. The reaction is continued while maintaining the temperature and stirring speed. The solution after the reaction is completed is centrifuged and the collected product is washed 3-5 times with ethanol and 3-5 times with deionized water to obtain uniformly dispersed solid silica nanospheres with uniform particle size.

[0015] Furthermore, the volume ratio of anhydrous ethanol, ammonia water, tetraethyl orthosilicate and deionized water is 50:(2-3):(1.8-2.0):(1.2-1.8), the stirring speed in step (1.1) is 1000-1200 rpm, the heating temperature is 60-70°C, the reaction time in step (1.2) is 6-7h, and the centrifugal speed is 16000-20000 rpm.

[0016] Furthermore, step (2) includes the following steps:

[0017] (2.1) Dissolve the etchants manganese sulfate monohydrate and disodium maleate in deionized water and mix well;

[0018] (2.2) adding the solid silica nanospheres obtained in step (1) to the solution obtained in step (2.1), and mixing by ultrasonication to fully disperse them;

[0019] (2.3) The solution obtained in (2.2) was transferred to a polytetrafluoroethylene-lined reactor and placed in an oven for reaction. The solution after the reaction was completed was centrifuged and the collected product was washed with anhydrous ethanol and deionized water 3-5 times each to obtain manganese-doped mesoporous silica hollow spheres.

[0020] Furthermore, the dosage ratio of manganese sulfate monohydrate, disodium maleate, solid silica nanospheres and deionized water is 1 mmol:0.625 mmol:(30-50 mg):(18-20 mL), the ultrasonic time in step (2.2) is 10-20 min, the reaction temperature in step (2.3) is 180-200 ° C, the reaction time is 10-14 h, and the centrifugal speed is 18000-20000 rpm.

[0021] Furthermore, step (3) includes the following steps:

[0022] (3.1) Prepare PBS solutions of isoniazid and sodium benzothiazolesulfinate respectively and mix them evenly;

[0023] (3.2) adding the manganese-doped mesoporous silica hollow spheres obtained in step (2) to the solution obtained in step (3.1) and dispersing them evenly;

[0024] (3.3) The solution obtained in (3.2) was stirred and reacted at room temperature in the dark. After the reaction, the solution was centrifuged and the collected product was washed 3-5 times with PBS solution to obtain functionalized manganese-doped mesoporous silica nanomaterials.

[0025] Furthermore, the dosage ratio of isoniazid, sodium benzothiazolesulfinate, manganese-doped mesoporous silica hollow spheres and PBS solution is 4 mg:2 mg:(1.5-2.5 mg):(0.8-1.2 mL), the stirring speed in step (3.3) is 500-600 r / min, the reaction time is 10-12 h, and the centrifugal speed is 15000-20000 rpm.

[0026] Furthermore, step (4) includes the following steps:

[0027] (4.1) Disperse mouse breast cancer cell membranes evenly in PBS solution and process the membrane material using a polycarbonate membrane through a microextruder for at least 10 extrusions.

[0028] (4.2) The functionalized manganese-doped mesoporous silica nanomaterial obtained in step (3) is uniformly dispersed in a PBS solution, mixed with an equal volume of the cell membrane material obtained in step (4.1), and processed through a micro-extruder using a polycarbonate membrane for at least 10 extrusions;

[0029] (4.3) The product obtained in step (4.2) was centrifuged and dispersed in PBS solution for storage.

[0030] Furthermore, the usage ratio of the cell membrane, the functionalized manganese-doped mesoporous silica nanomaterial and the PBS solution is 2 mg:(1.5-2.5 mg):(0.8-1.2 mL), the pore size of the polycarbonate membrane in step (4.1) is 400 nm, the pore size of the polycarbonate membrane in step (4.2) is 200 nm, the centrifugation speed in step (4.3) is 5000-7000 rpm, the time is 8-12 min, and the storage temperature is 0-10°C.

[0031] One of the technical solutions of the present invention is to provide a functionalized manganese-doped mesoporous nanomaterial and its application. The cell membrane-modified functionalized manganese-doped mesoporous silica nanomaterial (CCM@MIB) prepared by the method is used as a hydroxyl radical generator, a gas generator and a magnetic resonance imaging (MRI) contrast agent in the synergistic treatment of magnetic resonance imaging and tumors.

[0032] This material has targeting capabilities and good magnetic resonance imaging performance. It can generate hydroxyl radicals through non-Fenton reactions in the tumor microenvironment and is loaded with sulfur dioxide gas donors, which can ensure the selective accumulation of gas in specific areas while maintaining a low concentration in the blood circulation.

[0033] Isoniazid (INH), also known as isoniazid, is one of the most common anti-tuberculosis drugs. 2+ It can catalyze the conversion of INH to hydroxyl radicals (·OH) via a non-Fenton reaction, leading to increased levels of reactive oxygen species (ROS) in tumor cells. High-valent manganese can deplete reduced glutathione (GSH), which has ·OH scavenging properties, making it a promising candidate for improving ·OH-mediated cancer therapy. Furthermore, specific recognition of cancer cells and subcellular organelle-specific reagents can achieve efficient and specific accumulation at the tumor site, effectively addressing the problem of drug spillover, thereby improving treatment efficiency and reducing side effects.

[0034] Recently, the use of gaseous molecules such as nitric oxide (NO), carbon monoxide (CO), hydrogen (H2), and sulfur dioxide (SO2) to induce cancer cell death has made gas therapy superior to chemotherapy, radiotherapy, and other existing treatments due to their good diffusion in tumor tissues, the high permeability and long retention (EPR) effect, and the improved accumulation of nanoparticles at the tumor site. To make the gas molecules more controllable, gas prodrugs and gas donors have been designed to ensure the selective accumulation of gases in specific areas while maintaining low concentrations in the blood circulation.

[0035] SO₂ has long been considered a toxic environmental pollutant and a byproduct of industrial processing, but recent studies have demonstrated both toxic and protective effects in mammals. SO₂ can induce apoptosis while simultaneously upregulating intracellular reactive oxygen species (ROS) levels and modulating the expression of apoptosis-related proteins. SO₂ can also ameliorate multidrug resistance in tumor cells and possesses excellent cell penetration, making it suitable for the treatment of deep-seated tumors. To treat specific diseases, SO₂ must be precisely delivered to the target tissue. Several SO₂-delivering small-molecule prodrugs based on diverse release mechanisms, including GSH, pH, and light-induced release, have been developed. However, SO₂ delivery and release remain limited by the depth of penetration in vivo and in situ. Furthermore, once initiated, endogenous glutathione stimulation cannot be stopped or interrupted. Gas therapy using SO₂ prodrug-doped nanoparticles has the potential to inhibit not only superficial but also deep-seated tumors.

[0036] Therefore, the present invention uses manganese-doped hollow mesoporous silica as a carrier, simultaneously loading isoniazid and the sulfur dioxide prodrug benzothiazole sulfinate within the cavity. Finally, the surface of the hollow sphere is coated with cancer cell membranes to produce a nanomaterial with excellent biocompatibility. Magnetic resonance imaging of manganese ions can determine the optimal accumulation time of the material at the tumor site. Furthermore, coating the cancer cell membranes can increase the accumulation of the nanomaterial at the tumor site, thereby improving the therapeutic effect.

[0037] Compared with the prior art, the present invention has the following advantages:

[0038] (1) The present invention prepares manganese-doped hollow mesoporous silica spheres with uniform dispersion and uniform particle size, which can release Mn in the tumor microenvironment. 2+ , has a good magnetic resonance imaging effect; Mn 2+ It can catalyze the generation of ·OH from isoniazid through a non-Fenton reaction, leading to an increase in the content of reactive oxygen species in tumor cells. High-valent manganese can consume reduced glutathione with ·OH scavenging ability, making it a good choice for improving ·OH-mediated cancer therapy.

[0039] (2) The sulfur dioxide gas donor loaded by the present invention can continuously release SO2 in the tumor microenvironment, which can induce cell apoptosis, while upregulating the level of intracellular reactive oxygen species and regulating the expression of apoptosis-related proteins. SO2 can also improve the multidrug resistance of tumor cells and has excellent cell penetration ability, and can be used for the treatment of deep tumor tissues.

[0040] (3) The coating of the cancer cell membrane of the present invention can increase the accumulation of nanomaterials at the tumor site, thereby improving the therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 Flow chart of the preparation method of cell membrane modified functionalized manganese-doped mesoporous silica nanomaterial (CCM@MIB) in an embodiment of the present invention;

[0042] Figure 2 1 is a scanning electron microscope (SEM) image of solid silicon dioxide nanospheres (SiO2) in an embodiment of the present invention;

[0043] Figure 3 TEM image of SiO2 in an embodiment of the present invention;

[0044] Figure 4 : is a SEM image of manganese-doped mesoporous silica hollow spheres (MH) in an embodiment of the present invention;

[0045] Figure 5 TEM image of MH in an embodiment of the present invention;

[0046] Figure 6: is a nitrogen adsorption and desorption isotherm of MH in an embodiment of the present invention;

[0047] Figure 7 is a pore size distribution curve of MH in an embodiment of the present invention;

[0048] Figure 8 is an X-ray photoelectron spectrum (XPS) diagram of MH in an embodiment of the present invention;

[0049] Figure 9 This is the XPS spectrum peak fitting diagram of Mn 2p orbital in the embodiment of the present invention;

[0050] Figure 10 This is the UV-visible spectrum of CCM@MIB in the embodiment of the present invention;

[0051] Figure 11 This is a summary diagram of the zeta potential of CCM@MIB in an embodiment of the present invention;

[0052] Figure 12 This is a sulfur dioxide gas release curve of sodium benzothiazolesulfinate at the solution level prepared according to the literature in the examples of the present invention;

[0053] Figure 13 1 is the absorption spectrum of the reaction between functionalized manganese-doped mesoporous silica nanomaterials (MIB) and reduced glutathione at different concentrations in the embodiment of the present invention;

[0054] Figure 14 This is a graph showing the consumption capacity of reduced glutathione at a solution level for different concentrations of MIB in an embodiment of the present invention;

[0055] Figure 15 This is a graph showing the generation of hydroxyl radicals by MIB at different material concentrations according to an embodiment of the present invention;

[0056] Figure 16 This is a hydroxyl radical generation curve of MIB at pH 5.0 in an embodiment of the present invention;

[0057] Figure 17 This is a hydroxyl radical generation curve of MIB at pH 6.5 in an embodiment of the present invention;

[0058] Figure 18 This is a hydroxyl radical generation curve of MIB at pH 7.4 in an embodiment of the present invention;

[0059] Figure 19 This is a sulfur dioxide gas release curve of MIB at the solution level in an embodiment of the present invention;

[0060] Figure 201 / T1 and 1 / T2 are linear fitting curves of MIB and manganese ion concentration in an embodiment of the present invention;

[0061] Figure 21 Graphs showing changes in T1 and T2 imaging signals of MIB solutions with different concentrations according to an embodiment of the present invention;

[0062] Figure 22 This is a graph showing the cell viability after co-incubation of CCM@MIB and human umbilical vein endothelial cells in an embodiment of the present invention;

[0063] Figure 23 This is a graph showing the cell survival rate after co-incubation of cell membrane-modified manganese-doped mesoporous silica hollow spheres with mouse breast cancer cells in Comparative Example 1 of the present invention;

[0064] Figure 24 Graph showing cell survival rates after co-incubation of mouse breast cancer cells with different materials in Comparative Examples 1 to 3 and Examples of the present invention. DETAILED DESCRIPTION

[0065] The present invention is described in detail below with reference to specific embodiments. This embodiment is implemented based on the technical solution of the present invention, and provides a detailed implementation method and specific operation process, but the protection scope of the present invention is not limited to the following embodiments.

[0066] Unless otherwise specified, the equipment used in the following embodiments are all conventional equipment in the art; anything not described in detail in the following embodiments can be achieved by conventional experimental means in the art.

[0067] A composite nanomaterial for treating hydroxyl radicals and sulfur dioxide gas generated by a non-Fenton reaction guided by magnetic resonance imaging, in particular a functionalized manganese-doped mesoporous silica nanomaterial for cell membrane modification (CCM@MIB), and a preparation method and application thereof, such as Figure 1 As shown, the following steps are included:

[0068] (1) Manganese-doped mesoporous silica hollow spheres (MH) were synthesized by a hydrothermal method using manganese sulfate monohydrate and disodium maleate as etchants and solid silica nanospheres (SiO2) as templates;

[0069] (2) Isoniazid (INH) and sodium benzothiazolesulfinate (BTS) were loaded into the cavities and pores of manganese-doped mesoporous silica hollow spheres;

[0070] (3) Modifying the cell membrane (CCM) of mouse breast cancer cells (4T1 cells) on the surface of functionalized manganese-doped mesoporous silica nanomaterials (MIB) to improve the biocompatibility of the nanomaterials;

[0071] (4) Study the magnetic resonance imaging of the composite nanomaterial and the synergistic effect of hydroxyl radicals and sulfur dioxide gas treatment produced by non-Fenton reaction.

[0072] The raw materials used in the present invention are all commercially available raw materials or prepared by methods reported in existing public literature, such as:

[0073] Sodium benzothiazolesulfinate can be synthesized using the method reported in ACS Chem.Biol.2016,11,1647-1651.

[0074] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings.

[0075] Example:

[0076] 1. Solid silica nanospheres and their preparation method, the specific steps are as follows:

[0077] (1) Stir 50 mL of anhydrous ethanol, 2.5 mL of ammonia water, and 1.5 mL of deionized water in a three-necked flask at 1100 rpm and heat to 65°C.

[0078] (2) 1.9 mL of tetraethyl orthosilicate was added dropwise to the mixture of (1), stirred at 65° C. for 6.5 h, centrifuged at 18,000 rpm, and washed with anhydrous ethanol and deionized water three times each to obtain solid silica nanospheres with uniform particle size.

[0079] like Figure 2 and 3 As shown, the prepared solid silica nanospheres are evenly dispersed and have uniform particle size, with an average particle size of 90 nm.

[0080] 2. Manganese-doped mesoporous silica hollow spheres and preparation method thereof, the specific steps are as follows:

[0081] (1) Dissolve 169 mg of manganese sulfate monohydrate (MnSO4·H2O) and 92.5 mg of disodium maleate in 18 mL of deionized water and mix well.

[0082] (2) Add 30 mg of solid silica nanospheres to the mixture in (1) and sonicate for 15 min;

[0083] (3) The mixed solution of (2) was transferred to the polytetrafluoroethylene lining of a stainless steel reactor, placed in an oven at a temperature of 180°C for reaction for 12 h, centrifuged at 18,000 rpm, and washed with anhydrous ethanol and deionized water three times each to obtain manganese-doped mesoporous silica hollow spheres.

[0084] like Figure 4 and 5As shown, the hollow mesoporous silica nanospheres obtained after etching are evenly dispersed and have uniform particle size, with an average particle size of 93 nm.

[0085] like Figure 6 and 7 As shown, the specific surface area of ​​the nanosphere is 542.31m 2 / g, pore diameter of 6.57nm, pore volume of 0.89cm 3 / g.

[0086] like Figure 8 and 9 As shown, the strongest peaks at 654.5 eV and 642.7 eV are attributed to Mn 3+ 2p 1 / 2 and 2p 3 / 2 , two additional peaks were observed at 659.9 eV and 648.0 eV, indicating the coexistence of Mn(IV).

[0087] 3. Functionalized manganese-doped mesoporous silica nanomaterial and its preparation method, the specific steps are as follows:

[0088] (1) Prepare 10 mL of a 4 mg / mL isoniazid solution in PBS and 10 mL of a 2 mg / mL sodium benzothiazolesulfinate solution in PBS and mix them evenly;

[0089] (2) adding 20 mg of manganese-doped mesoporous silica hollow spheres to the mixture of (1) and dispersing them evenly;

[0090] (3) The mixture of (2) was stirred at a speed of 600 r / min at room temperature in the dark for 12 h, centrifuged at 18000 rpm, and washed three times with PBS solution to obtain functionalized manganese-doped mesoporous silica nanomaterials.

[0091] 4. Cell membrane modified functional manganese-doped mesoporous silica nanomaterial and its preparation method, the specific steps are as follows:

[0092] (1) The cell membranes of mouse breast cancer cells (4T1 cells) were uniformly dispersed in PBS solution at a concentration of 2 mg / mL. 0.5 mL of the cell membrane material was processed using a 400 nm polycarbonate membrane through a micro-extruder and squeezed 10 times.

[0093] (2) Functionalized manganese-doped mesoporous silica nanomaterials were uniformly dispersed in PBS solution at a concentration of 2 mg / mL, mixed with equal volumes of the cell membrane material (1), and processed using a 200 nm polycarbonate membrane through a micro-extruder and extruded 10 times;

[0094] (3) The product of (2) was centrifuged at 6000 rpm for 10 min, dispersed in PBS solution and stored at 4° C. to obtain functionalized manganese-doped mesoporous silica nanomaterials modified with cell membranes.

[0095] like Figure 10 As shown, MH has no characteristic absorption peak in the UV-visible region. Isoniazid has a distinct absorption peak at 262 nm. Sodium benzothiazolesulfinate has two distinct absorption peaks at 262 nm and 294 nm. Cancer cell membrane has a sharp absorption peak at 222 nm and a peak around 275 nm. MIB has an absorption peak at 262 nm and 294 nm. CCM@MIB has three peaks at 225 nm, 275 nm, and 294 nm, corresponding to the UV-visible absorption peaks of cancer cell membrane, isoniazid, and sodium benzothiazolesulfinate.

[0096] like Figure 11 As shown in Figure 3, the changes in Zeta potential can prove that the material in each step is successfully prepared.

[0097] like Figure 12 As shown in the figure, with the decrease of pH and the extension of time, the release of sulfur dioxide from sodium benzothiazolesulfinate gradually increased, and slow and continuous release could be achieved under 24h continuous monitoring.

[0098] like Figure 13 and 14 As shown in the figure, as the concentration of MIB increases, the consumption capacity of reduced glutathione gradually increases. The consumption of 5 mM reduced glutathione by 400 μg / mL MIB is 93.5%. The experimental results show that MIB has a good consumption capacity for reduced glutathione.

[0099] like Figures 15 to 18 As shown, using tetramethylbenzidine (TMB) as a hydroxyl radical indicator, the hydroxyl radical generation effect of MIB was measured by UV-visible absorption spectroscopy. Under the same conditions, the absorbance of the tetramethylbenzidine solution increased with increasing MIB concentration. Furthermore, the hydroxyl radical generation effect of MIB was affected by pH. Under neutral conditions, the absorbance of the solution at 652 nm did not increase significantly, indicating that MIB generates almost no hydroxyl radicals under neutral conditions, thus avoiding damage to normal cells. However, under slightly acidic conditions, the absorbance of the solution at 652 nm increased significantly, indicating that MIB has a strong ability to generate hydroxyl radicals in slightly acidic environments.

[0100] like Figure 19As shown, similar to the simple sulfur dioxide gas donor sodium benzothiazolesulfinate, the release of MIB sulfur dioxide gradually increased with the decrease of pH and the extension of time. Sulfur dioxide gas was almost not released at pH = 7.4, but in the tumor microenvironment of tumor cells (pH = 6.5), slow and continuous release could be achieved under 24h continuous monitoring.

[0101] like Figure 20 and 21 As shown in the results, MIB has a good T1-weighted MRI effect and can be used as a T1-weighted MRI contrast agent.

[0102] like Figure 22 As shown, after co-incubation with human umbilical vein endothelial cells for 12h and 24h, CCM@MIB had no obvious cytotoxicity and high biocompatibility, and could be used for in vivo treatment.

[0103] Comparative Example 1:

[0104] A cell membrane modified manganese-doped mesoporous silica hollow sphere (CCM@MH) and a preparation method thereof are basically the same as those in the embodiment, except that isoniazid and sodium benzothiazolesulfinate are not loaded.

[0105] Comparative Example 2:

[0106] A cell membrane modified functionalized manganese-doped mesoporous silica nanomaterial (CCM@MI) and a preparation method thereof are basically the same as those in the embodiment, except that only isoniazid is loaded.

[0107] Comparative Example 3:

[0108] A cell membrane modified functionalized manganese-doped mesoporous silica nanomaterial (CCM@MB) and a preparation method thereof are basically the same as those in the embodiment, except that only sodium benzothiazolesulfinate is loaded.

[0109] like Figure 23 As shown in Figure 2, after CCM@MH was co-incubated with mouse breast cancer cells (4T1 cells) for 12 h and 24 h, the cell survival rate gradually decreased with the increase of carrier material concentration, indicating that the material has a certain therapeutic effect; Figure 24 As shown in the figure, the cell survival rate of CCM@MIB was lower than that of the simple vehicle, which decreased the cell survival rate to 13.2%, indicating that the therapeutic effect of CCM@MIB was obvious.

[0110] The preparation method of the present invention is simple, and the resulting composite nanomaterial has magnetic resonance imaging contrast performance and good synergistic therapeutic effects. This method can be used for magnetic resonance imaging-guided tumor treatment, achieving integrated diagnosis and treatment.

[0111] The above description of the embodiments is intended to facilitate understanding and use of the invention by those skilled in the art. It will be apparent that those skilled in the art can readily make various modifications to these embodiments and apply the general principles described herein to other embodiments without requiring inventive effort. Therefore, the present invention is not limited to the above-described embodiments. Improvements and modifications made by those skilled in the art based on the disclosure of the present invention, without departing from the scope of the present invention, should be within the scope of protection of the present invention.

Claims

1. A method for preparing a cell membrane modified functional manganese-doped mesoporous silica nanomaterial, characterized in that: The method comprises the following steps: (1) Preparation of solid silica nanospheres; (2) Mn-doped mesoporous silica hollow spheres were prepared by mixing solid silica nanospheres with maleate and manganese salt aqueous solutions using a hydrothermal method; (3) Isoniazid and benzothiazole sulfinate were loaded into manganese-doped mesoporous silica hollow spheres to obtain functionalized manganese-doped mesoporous silica nanomaterials; (4) The cell membrane of the treated mouse breast cancer cells was composited with the functionalized manganese-doped mesoporous silica nanomaterial to prepare the cell membrane-modified functionalized manganese-doped mesoporous silica nanomaterial; Step (3) includes the following steps: (3.1) Prepare PBS solutions of isoniazid and sodium benzothiazolesulfinate respectively and mix them; (3.2) adding the manganese-doped mesoporous silica hollow spheres obtained in step (2) to the solution obtained in step (3.1); (3.3) The solution obtained in (3.2) was stirred and reacted at room temperature in the dark. After the reaction, the solution was centrifuged to obtain functionalized manganese-doped mesoporous silica nanomaterials.

2. The method for preparing a cell membrane modified functional manganese-doped mesoporous silica nanomaterial according to claim 1, characterized in that: Step (1) includes the following steps: (1.1) Stir and mix ethanol, ammonia water, and water, and heat; (1.2) When the temperature is stable, tetraethyl orthosilicate is added to the solution obtained in step (1.1), and the reaction is continued. After the reaction is completed, the solution is centrifuged to obtain solid silica nanospheres.

3. The method for preparing a cell membrane modified functional manganese-doped mesoporous silica nanomaterial according to claim 2, characterized in that: The volume ratio of ethanol, ammonia water, tetraethyl orthosilicate and water is 50:(2-3):(1.8-2.0):(1.2-1.8). The stirring speed in step (1.1) is 1000-1200 rpm, the heating temperature is 60-70 ° C, the reaction time in step (1.2) is 6-7 h, and the centrifugal speed is 16000-20000 rpm.

4. The method for preparing a cell membrane modified functional manganese-doped mesoporous silica nanomaterial according to claim 1, characterized in that: Step (2) includes the following steps: (2.1) Dissolve manganese sulfate monohydrate and disodium maleate in water; (2.2) adding the solid silica nanospheres obtained in step (1) to the solution obtained in step (2.1) and mixing by ultrasonication; (2.3) The solution obtained in (2.2) is reacted, and the solution after the reaction is centrifuged to obtain manganese-doped mesoporous silica hollow spheres.

5. The method for preparing a cell membrane modified functional manganese-doped mesoporous silica nanomaterial according to claim 4, characterized in that: The dosage ratio of manganese sulfate monohydrate, disodium maleate, solid silica nanospheres and water is 1 mmol:0.625 mmol:(30-50 mg):(18-20 mL). The ultrasonic time in step (2.2) is 10-20 min. The reaction temperature in step (2.3) is 180-200 °C, the reaction time is 10-14 h, and the centrifugal speed is 18000-20000 rpm.

6. The method for preparing a cell membrane modified functional manganese-doped mesoporous silica nanomaterial according to claim 1, characterized in that: The dosage ratio of isoniazid, sodium benzothiazolesulfinate, manganese-doped mesoporous silica hollow spheres, and PBS solution is 4 mg:2 mg:(1.5-2.5 mg):(0.8-1.2 mL). The stirring speed in step (3.3) is 500-600 r / min, the reaction time is 10-12 h, and the centrifugal speed is 15000-20000 rpm.

7. The method for preparing a cell membrane modified functional manganese-doped mesoporous silica nanomaterial according to claim 1, characterized in that: Step (4) includes the following steps: (4.1) Disperse mouse breast cancer cell membranes in PBS solution and treat the cell membrane material with a polycarbonate membrane; (4.2) dispersing the functionalized manganese-doped mesoporous silica nanomaterial obtained in step (3) in a PBS solution, mixing it with an equal volume of the cell membrane material obtained in step (4.1), and treating it with a polycarbonate membrane; (4.3) Centrifuge the product obtained in step (4.2) and disperse it in PBS solution for storage.

8. The method for preparing a cell membrane modified functional manganese-doped mesoporous silica nanomaterial according to claim 7, characterized in that: The dosage ratio of cell membrane, functionalized manganese-doped mesoporous silica nanomaterial and PBS solution is 2 mg:(1.5-2.5 mg):(0.8-1.2 mL), the pore size of the polycarbonate membrane in step (4.1) is 400 nm, the pore size of the polycarbonate membrane in step (4.2) is 200 nm, the centrifugation speed in step (4.3) is 5000-7000 rpm, the time is 8-12 min, and the storage temperature is 0-10 ℃.

9. Use of the cell membrane-modified functionalized manganese-doped mesoporous silica nanomaterial prepared by the method according to any one of claims 1 to 8 in the preparation of a drug for treating breast cancer.

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