Mn / MoO3-X nano material as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of nano medicines, and discloses a Mn / MoO3-X nano material as well as a preparation method and application thereof. The material is prepared by the following steps: adding a molybdenum source, fatty amine and a surface stabilizer into a high-boiling-point solvent at normal pressure and 60-120 DEG C to prepare a MoO3 matrix, then adding a manganese source, stirring and dispersing, then transferring to a high-pressure kettle, carrying out solvothermal reaction for 6-24 hours at 100-280 DEG C, treating to obtain an oil-phase Mn / MoO3-X nano material, and then carrying out surface modification to obtain the oil-phase Mn / MoO3-X nano material. The hydrophobic surface of the oil-phase Mn / MoO3-X nano material is converted into a hydrophilic surface, so that the oil-phase Mn / MoO3-X nano material has good colloidal stability. According to the nanometer material, heterogeneous manganese single atom doping is achieved on the basis of a MoO3-x matrix rich in defects, the activation efficiency and continuity of a cGAS / STING pathway are remarkably improved, and the nanometer material has good enzyme-like catalytic performance and biological safety, can remodel the microenvironment of tumor site immunosuppression and has a good specific metal immunotherapy effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of nanomedicine and specifically relates to a Mn / MoO 3-X Nanomaterials and their preparation methods and applications. Background Art

[0002] Cancer, as one of the killers of human health, has long threatened people's lives and health. Immunotherapy has brought revolutionary changes to cancer treatment and provided effective treatment options for different types of tumors. Despite the success of immunotherapy, only a few cancer patients currently show good treatment effects, a phenomenon that is mainly attributed to the powerful immunosuppressive network in the tumor microenvironment. Cyclic-GMP-AMP (cGAMP) synthetase (cGAS) can recognize double-stranded DNA in the cell membrane, induce the production of cGAMP, activate the stimulator of interferon genes (STING), and thus initiate an anti-tumor immune response. To date, a variety of STING agonists have shown good disease treatment effects. However, they still face many challenges in clinical application, such as short half-life in vivo, low bioavailability, and easy nonspecific and adverse reactions.

[0003] Catalytic therapy has attracted wide attention as an emerging strategy for tumor treatment. It kills tumor cells by converting non-toxic or low-toxic endogenous substances into highly toxic molecules, thereby improving the therapeutic effect and reducing side effects. The tumor microenvironment is very different from the normal physiological environment, including lower pH, high levels of GSH, high levels of H2O2, etc. Nanozymes with POD / OXD enzyme-like activity can catalyze the high levels of H2O2 in the tumor microenvironment to produce ROS, such as singlet oxygen ( 1 O2), superoxide anion (·O2 - ) and hydroxyl radicals (·OH), etc. ROS induce tumor cell death by destroying intracellular biomolecules (such as proteins, lipids, and DNA).

[0004] The manganese element of manganese-based nanozymes has a variety of different oxidation states. The manganese atoms in low oxidation states can exhibit enzyme-like catalytic activity to produce ROS, while the manganese atoms in higher oxidation states can be reduced by glutathione (GSH) and release Mn 2+ The generated ROS can promote the breakage and leakage of double-stranded DNA and react with the released Mn 2+ Synergistically activate the cGAS / STING signaling pathway to achieve combined catalytic therapy for tumor immunotherapy. However, the enzyme-like catalytic performance of the currently reported manganese-based nanozymes is not ideal, and the ability to induce ROS production is weak; in addition, membrane transport proteins actively expel Mn 2+ , resulting in Mn 2+The concentration is reduced, thus limiting the activation effect of the cGAS / STING pathway, and ultimately resulting in the inability of existing manganese-based nanozymes to achieve good anti-tumor therapeutic effects. It can be seen that there is an urgent need to develop a nanomaterial that can efficiently generate a large amount of ROS, has high tumor cell killing efficiency, and has good in vivo biosafety to sustainably activate the cGAS / STING pathway in order to enhance the anti-tumor effect.

[0005] In recent years, single-atom nanomaterials have made great achievements in catalysis and biomedicine due to their advantages such as high atomic utilization and strong metal-matrix interface interaction. The main methods for constructing single-atom nanomaterials include defect engineering strategy, spatial confinement strategy, and designed coordination strategy. Studies have found that molybdenum (MoO4 2- ) as an essential trace element also has the ability to activate the cGAS-STING pathway and is more effective than Mn 2+ Therefore, the inventors speculated that based on the team's previous research on molybdenum oxide and defect engineering strategies, it is expected that the anchoring of manganese atoms can be achieved through the coordination of metal single atoms with defect sites, which is expected to improve the efficiency of ROS generation and make up for the Mn 2+ Too rapid efflux results in a short-lived activation efficiency of cGAS / STING. Summary of the invention

[0006] In order to solve the above problems, the present invention provides a Mn / MoO 3-X Nanomaterials. The material is based on defect-rich MoO 3-x The matrix can achieve heterogeneous Mn single atom doping, which can not only achieve strong peroxidase mimic catalysis and induce severe double-stranded DNA damage in tumors, but also 2+ and MoO4 2- It can be transformed from Mn / MoO in the tumor microenvironment 3-x The slow release from nanomaterials to achieve long-term cascade activation of the cGAS / STING pathway can be used for tumor-specific catalytic metal immunotherapy, providing a feasible solution for the development of new tumor immunotherapy drugs.

[0007] The technical solution of the present invention is as follows:

[0008] The present invention provides a Mn / MoO 3-X The method for preparing a nano material comprises the following steps:

[0009] (1) Adding a molybdenum source, a fatty amine and a surface stabilizer to a high boiling point solvent, stirring at 60 to 120° C. for 5 to 60 minutes, then adding a manganese source, and continuing to stir at 60 to 120° C. for 1 to 60 minutes; then transferring the mixture to a polytetrafluoroethylene autoclave, performing a solvent thermal reaction at 100 to 280° C. for 6 to 24 hours, washing with an organic polar solvent, centrifuging, and dispersing in an organic non-polar solvent to obtain an oil phase Mn / MoO 3-X Nanomaterial; the mass volume ratio of the high boiling point solvent, the molybdenum source, the fatty amine, the surface stabilizer and the manganese source is 4-12 mL: 4-400 mg: 20-2000 mg: 0.5-10 mL: 4-400 mg;

[0010] (2) Using a surface modification method, the oil phase Mn / MoO 3-X The nanomaterial and the modified molecule are dissolved in the organic non-polar solvent, stirred at 20-30°C for 0-24h, and then rotary evaporated, and then distilled water is added to disperse to obtain Mn / MoO 3-X Nanomaterials.

[0011] In one embodiment of the present invention, in step (1), the high boiling point solvent is preferably one or more of octadecene, benzyl ether, ethylene glycol, N,N-dimethylformamide or dimethyl sulfoxide.

[0012] In one embodiment of the present invention, in step (1), the molybdenum source is preferably one or more of molybdenum acetylacetonate, molybdenum acetate, sodium molybdate, carbonyl molybdenum or molybdenum chloride.

[0013] In one embodiment of the present invention, in step (1), the surface stabilizer is preferably one or more of oleic acid, oleylamine or oleyl alcohol; the fatty amine is preferably one or more of hexadecylamine, dodecylamine, tetradecylamine or octadecylamine. Oleic acid and fatty amine can slowly generate water molecules in a solvent thermal system, which is beneficial to accelerate the in-situ dissolution of the MoO3 matrix and the formation of surface defect vacancies.

[0014] In one embodiment of the present invention, in step (1), the manganese source is preferably one or more of manganese acetylacetonate, manganese oleate, manganese acetate or manganese pentacarbonyl.

[0015] In one embodiment of the present invention, the organic non-polar solvent is preferably one or more of dichloromethane, chloroform, n-hexane, cyclohexane, dioxane, o-dichlorobenzene or toluene, and the organic polar solvent is one or more of acetone, ethanol, ethyl acetate, methanol, methyl pyrrolidone, medium-chain alcohol, acetonitrile, N,N-dimethylformamide or dimethyl sulfoxide.

[0016] In one embodiment of the present invention, in step (2), the modified molecule is preferably one or more of phospholipid-polyethylene glycol, polyethylene glycol, pluronic, polyvinyl pyrrolidone, polyvinyl alcohol, Tween, vitamin E polyethylene glycol succinate, polylactic acid-co-glycolic acid, polydopamine, meso-dithiosuccinate, cysteine, mercaptosuccinic acid, citric acid or mercaptopropionic acid. The modified molecule can be Mn / MoO 3-X The hydrophobic surface of the nanomaterial is transformed into a hydrophilic surface, giving it good colloidal stability.

[0017] The present invention also discloses a method comprising the above-mentioned Mn / MoO 3-X Mn / MoO prepared by the preparation method of nanomaterials 3-X Nanomaterials.

[0018] The present invention also discloses the above-mentioned Mn / MoO 3-X Application of nanomaterials in the preparation of anti-tumor therapeutic drugs.

[0019] In one embodiment of the present invention, the Mn / MoO 3-X Nanomaterials slowly release Mn 2+ and MoO4 2- Long-term cascade activation of the cGAS / STING pathway.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] A Mn / MoO 3-X The MoO3 matrix was first prepared under normal pressure heating conditions, and then the in-situ defect-rich MoO3 was obtained by aliphatic amine-mediated reduction and in-situ dissolution under solvothermal reaction conditions. 3-x The matrix (defect site is the anchoring point of single manganese atom) can achieve efficient adsorption and anchoring of heterogeneous manganese metal, realize single atom doping of heterogeneous metal, and then obtain nanomaterials that are stably dispersed in water through surface modification. The nanomaterial can achieve powerful peroxidase mimic catalysis and induce severe double-stranded DNA (dsDNA) damage in tumors; in addition, Mn / MoO 3-x Nanomaterials responsively release Mn in high-level GSH environment 2+ and MoO4 2- , increasing the sensitivity of cGAS to dsDNA. More importantly, MoO4 2- The release rate of Mn is relatively slow and it is easy to accumulate in cells, which makes up for the 2+The lack of cell accumulation caused by continuous outflow can continuously trigger cells to secrete type I interferon for up to 72 hours. Therefore, the long-term cascade activation of the cGAS / STING pathway by the nanomaterials of the present invention can be used for tumor-specific catalytic metal immunotherapy, providing a feasible solution for the development of new tumor immunotherapy drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 A Mn / MoO 3-X Transmission electron microscopy (TEM) images of nanomaterials;

[0023] Figure 2 A Mn / MoO 3-X Elemental analysis (EDS-Mapping) diagram of nanomaterials;

[0024] Figure 3 A Mn / MoO 3-X High-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) images of nanomaterials;

[0025] Figure 4-5 In Test Example 1, the Mn / MoO 3-X Schematic diagram of the in vitro POD enzyme-like catalytic performance of the nanomaterial;

[0026] Figure 6 In Test Example 2, the Mn / MoO 3-X Schematic diagram of the effect of nanomaterials on B16 cell viability;

[0027] Figure 7 In Test Example 3, the Mn / MoO 3-X Nanomaterials in B16 cells ions (Mn 2+ / MoO4 2- ) Release diagram;

[0028] Figure 8 In Test Example 4, the Mn / MoO 3-X Schematic diagram of the effect of nanomaterials on double-stranded DNA damage in B16 cells;

[0029] Fig. 9 In Test Example 5, the Mn / MoO 3-X Schematic diagram of the effect of nanomaterials on the activation of cGAS / STING pathway in B16 cells;

[0030] Fig.10 In Application Example 1, the Mn / MoO 3-X Schematic diagram of the results of nanomaterials inhibiting tumor growth;

[0031] Fig.11 In Application Example 2, the Mn / MoO 3-X Schematic diagram of the results of nanomaterials inhibiting tumor metastasis;

[0032] Fig.12 In Application Example 2, the Mn / MoO 3-X Schematic diagram of the effect of nanomaterials on dendritic cells in tumors;

[0033] Fig.13 In Application Example 2, the Mn / MoO 3-X Nanomaterials for CD4 + Schematic diagram of the impact of T cells;

[0034] Fig.14 In Application Example 2, the Mn / MoO 3-X Nanomaterials for CD8 in tumors + Schematic diagram of the effects of T cells. DETAILED DESCRIPTION

[0035] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below with reference to the accompanying drawings.

[0036] Many specific details are set forth in the following description to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways than those described herein, and those skilled in the art can make similar generalizations without violating the connotation of the present invention, so the present invention is not limited to the specific implementation disclosed below.

[0037] A Mn / MoO 3-X The nanomaterial, the preparation method thereof comprises the following steps:

[0038] (1) First, a molybdenum source, a fatty amine and a surface stabilizer are added to a high boiling point solvent, and the mixture is stirred at 60 to 120° C. for 5 to 60 minutes. Then, a manganese source is added, and the mixture is stirred at 60 to 120° C. for 1 to 60 minutes. The mixture is then transferred to a polytetrafluoroethylene autoclave, and a solvent thermal reaction is carried out at 100 to 280° C. for 6 to 24 hours. The mixture is then washed with an organic polar solvent, centrifuged, and dispersed in an organic non-polar solvent to obtain an oil phase Mn / MoO 3-X Nanomaterial; the mass volume ratio of the high boiling point solvent, the molybdenum source, the fatty amine, the surface stabilizer and the manganese source is 4-12 mL: 4-400 mg: 20-2000 mg: 0.5-10 mL: 4-400 mg;

[0039] (2) Using a surface modification method, the oil phase Mn / MoO 3-XThe nanomaterial and the modified molecule are dissolved in the organic non-polar solvent, stirred at 20-30°C for 0-24h, and then rotary evaporated, and then distilled water is added to disperse to obtain Mn / MoO 3-X Nanomaterials.

[0040] The present invention first synthesizes the MoO3 matrix under normal pressure heating conditions, and then obtains the MoO3 matrix rich in defect sites through aliphatic amine-mediated reduction and in-situ dissolution under solvent thermal reaction conditions. 3-x Matrix, in which MoO 3-x The molybdenum sites of the matrix can serve as anchoring sites for manganese single atoms, capturing and anchoring metallic manganese single atoms to achieve heterogeneous metallic manganese single atom doping.

[0041] Example 1

[0042] A Mn / MoO 3-X The nanomaterial, the preparation method thereof comprises the following steps:

[0043] (1) First, 40 mg of molybdenum acetylacetonate, 200 mg of hexadecylamine and 2.0 mL of oleic acid were added to 8 mL of octadecene, and then uniformly heated at 80 ° C for 10 min; 40 mg of manganese acetylacetonate was added to the above system and stirred at 80 ° C for 5 min; the mixture was transferred to a polytetrafluoroethylene autoclave (15 mL) and subjected to solvothermal reaction at 160 ° C for 12 h; the product was washed with ethanol and centrifuged, repeated 3 times, and dispersed in chloroform to obtain an oil phase Mn / MoO 3-X Nanomaterials;

[0044] The prepared oil phase Mn / MoO 3-X The morphology of nanomaterials was characterized by TEM. Figure 1 As shown, it is clearly shown that Mn / MoO 3-x The nanomaterial is a spherical structure with an average size of about 15nm. EDS-Mapping was used to analyze the Mn / MoO 3-X The distribution of elements in nanomaterials, such as Figure 2 As shown in Figure 2, molybdenum, manganese and oxygen are evenly distributed in the Mn / MoO 3-x Nanomaterials. Confirmation of Mn / MoO by HAADF-STEM 3-X The dispersed state of manganese atoms in nanomaterials, such as Figure 3 As shown, the dark spots (yellow circles) are scattered manganese atomic sites, and their neighboring molybdenum atoms appear brighter due to their higher atomic number.

[0045] (2) Weigh 20 mg of phospholipid-polyethylene glycol (DSPE-PEG 2000 ) and 2 mg oil phase Mn / MoO 3-XThe nanomaterial was dissolved in 5 mL of chloroform. Stirred at room temperature for 4 h, rotary evaporated at 60 °C for 1 h, and dispersed in distilled water to obtain aqueous Mn / MoO 3-X Nanomaterials.

[0046] Test Example 1

[0047] This test example 1 provides the Mn / MoO prepared in Example 1. 3-X The nanomaterials were tested for in vitro POD-like enzyme activity, and the test method was as follows:

[0048] MoO 3-x Nanoparticles (NPs), Mn / MoO 3-x The nanomaterials were added to an acetic acid / sodium acetate buffer solution (0.2 M, pH 5.0) to a final concentration of 10 μg / mL in a total volume of 2 mL. Then, different concentrations of hydrogen peroxide (H2O2) (20, 40, 80, and 100 μM) were added, and 20 μL of 3,3′,5,5′-tetramethylbenzidine (TMB) (1 mM) was added, and the absorbance at 652 nm was immediately measured using a UV-visible spectrophotometer.

[0049] like Figure 4-5 As shown, Mn / MoO 3-x The characteristic absorbance of oxidized TMB (oxTMB) of the nanomaterial at 652 nm was significantly increased, while MoO 3-x There was little change in the NPs group.

[0050] Test Example 2

[0051] This test example 2 provides the Mn / MoO prepared in Example 1. 3-X The cytotoxicity of nanomaterials was tested using the following test methods:

[0052] B16 cells (6×10 3 The cells were inoculated into 96-well plates and cultured in a 37°C cell culture incubator for 12 h. 3-XAfter the cells were incubated with the nanomaterial-containing amino acid and glucose medium (Dulbecco's Modified Eagle Medium, DMEM) for 24 hours, 100 μL of 10% Cell Counting Kit-8 (CCK-8) / DMEM solution was added to each well to replace the DMEM medium, and the cells were incubated in a 37°C cell culture incubator for 2 hours. Finally, the absorbance of each well at 450 nm was detected by an enzyme reader.

[0053] like Figure 6 As shown, Mn / MoO 3-X Nanomaterials can effectively induce cytotoxicity, IC 50 About 6.8μg / mL.

[0054] Test Example 3

[0055] This test example 3 provides the Mn / MoO prepared in Example 1. 3-X Ions (Mn) of nanomaterials in B16 cells 2+ / MoO4 2- ) Release capability test, the test method is as follows:

[0056] B16 cells (1×10 6 Cells were inoculated into 6-well plates and cultured in a 37°C cell culture incubator for 24 h. 3-x Fresh DMEM containing nanomaterials (5 μg / mL) replaced the original culture medium and incubated with cells for different time periods (0 h, 24 h, and 72 h). After the incubation, the cells were centrifuged (1000 rpm, 5 min), collected, and counted. The cells were then lysed with cell lysis buffer, and the concentrations of manganese and molybdenum in the cell lysate were determined by ICP-MS.

[0057] like Figure 7 As shown, intracellular Mn 2+ The concentration reached a peak at 24 hours and then dropped sharply. 2- The gradual accumulation of Mn 2+ The activation efficiency is reduced, thereby allowing the cascade activation of the cGAS / STING pathway to continue.

[0058] Test Example 4

[0059] This test example 4 provides a method for treating the Mn / MoO 3-X The effect of nanomaterials on cell double-strand DNA damage was tested using the following method:

[0060] B16 cells (1×10 5100 cells / dish) were inoculated into the confocal culture dish and cultured in a 37°C cell culture incubator for 24 h. Then, DMEM was replaced with Mn / MoO 3-x Nanomaterials (10 μg / mL) were added to fresh DMEM and cultured for 6 h, 12 h, and 24 h, respectively. The cells were washed twice with PBS and fixed with 4% paraformaldehyde solution for 15 min. Then, 1 mL of 0.5% Triton X-100 was used for permeabilization for 20 minutes and rinsed three times with PBS. 1 mL of 1% bovine serum albumin (BSA) was added to each culture dish for blocking for 30 min. Then, the cells were rinsed three times with PBS containing Tween 20, and then incubated at 37 ° C for 1 h with diluted anti-rabbit Alexa fluorescent 488 solution (Wuhan Bost Biotechnology Co., Ltd.), and finally observed with a laser confocal microscope.

[0061] like Figure 8 As shown in the immunofluorescence experiment, in Mn / MoO 3-x The nanomaterials group observed the generation of obvious γ-H2AX fluorescence signals, confirming that Mn / MoO 3-x Nanomaterials can significantly induce double-stranded DNA damage.

[0062] Test Example 5

[0063] This test example 5 provides the Mn / MoO prepared in Example 1. 3-X The effect of nanomaterials on the activation of the cGAS / STING pathway in cells was tested using the following method:

[0064] B16 cells (1×10 5 The cells were inoculated into 6-well plates and cultured in a 37°C cell culture incubator for 24 h. Then, DMEM was replaced with PBS, MoO 3-x NPs (10 μg / mL), Mn 2+ (10 μg / mL) and Mn / MoO 3-xNanomaterials (10 μg / mL) were added to fresh DMEM. After 24 h, the cells were collected by centrifugation (1000 rpm, 5 min) and lysed with cell lysis buffer. The total protein concentration was quantitatively detected using a BCA protein assay kit (Thermo Fisher Scientific, USA). At the same time, equal amounts of protein (about 20 μg) were separated by SDS polyacrylamide gel and electrotransferred to polyvinylidene fluoride (PVDF) membranes. Then, the membranes were blocked with 5% skim milk and incubated overnight at 4 ° C with specific primary antibodies (including β-actin, TANK binding kinase 1 (TBK1), phosphorylated-TBK1 (P-TBK1), interferon regulatory factor 3 (IRF3), phosphorylated-IRF3 (P-IRF3), interferon gene stimulator (STING), phosphorylated-STING (P-STING)) (Cell Signaling Technology, USA), and then incubated with the corresponding secondary antibodies. Finally, enhanced chemiluminescence (ECL) substrate (Thermo Fisher Scientific, USA) was used for chemiluminescence detection to display protein bands.

[0065] like Fig. 9 As shown, in the Mn / MoO 3-x In B16 cells treated with nanomaterials, the expression levels of phosphorylated STING (p-STING), phosphorylated TBK1 (p-TBK1), and phosphorylated IRF3 (p-IRF3) were significantly increased, significantly exceeding those in B16 cells treated with MoO 3-x NPs or Mn 2+ Treatment group.

[0066] Application Example 1

[0067] This application example 1 provides the Mn / MoO 3-X The application test of nanomaterials on tumor inhibition in vivo is as follows:

[0068] Each mouse was injected with B16 cells (2×10 5 When the tumor volume reaches about 100mm 3 C57BL / 6 mice bearing B16 tumors were randomly divided into four groups (n=5) and injected intravenously with PBS, MoO 3-x NPs (5 mg / kg), Mn 2+ (5mg / kg) and Mn / MoO 3-x Nanomaterials (5 mg / kg) were added, and the tumor volume was recorded using a vernier caliper every two days.

[0069] like Fig.10 As shown, the embodiment of the present invention 1Mn / MoO3-X Nanomaterials can significantly inhibit tumor growth, and the inhibitory effect is stronger than MoO 3-x Nanomaterials Group.

[0070] Application Example 2

[0071] This application example 2 provides the Mn / MoO 3-X The application test of nanomaterials in inhibiting tumor metastasis and remodeling the tumor immune microenvironment is as follows:

[0072] Each mouse was injected with B16 cells (2×10 5 Three days later, B16 tumor cells (2×10 5 / mouse) to establish a bilateral subcutaneous tumor model. When the left tumor volume reached about 100mm 3 C57BL / 6 mice bearing B16 tumors were randomly divided into four groups (n=5) and injected intratumorally with PBS, MoO 3-x NPs (5 mg / kg), Mn 2+ (5mg / kg) and Mn / MoO 3-x Nanomaterials (5 mg / kg). After the treatment on the 12th day, tumor tissues were obtained and treated with 0.6 mg / mL collagenase IV and 0.01 mg / mL deoxyribonuclease I (DNaseI). After digestion in culture medium at 37°C for 1 hour, the cells were filtered using a 40 μm cell filter and then treated with 36% Percoll layering solution and lysed red solution, respectively. The single cell suspension was incubated with Fc blockers, and then the cells isolated from the tumor were stained with the corresponding antibodies. Flow cytometry was used to analyze the expression of dendritic cells and CD8 in tumor tissues. + T cells and CD4 + T cell changes.

[0073] like Figure 11-Figure 14 As shown, Mn / MoO 3-X Nanomaterials can promote the maturation of dendritic cells in tumor tissues, facilitate subsequent antigen presentation, and increase the expression of CD8 + T cells and CD4 + At the T cell level, it ultimately prevented tumor metastasis, successfully inhibited the growth of distant tumors, and exerted a highly effective anti-tumor specific metal immunotherapy effect.

[0074] Example 2

[0075] A Mn / MoO 3-X The nanomaterial, the preparation method thereof comprises the following steps:

[0076] First, 40 mg of molybdenum acetylacetonate, 200 mg of hexadecylamine and 2.0 mL of oleic acid were added to 8 mL of octadecene, and then uniformly heated at 80 ° C for 10 min; 400 mg of manganese acetylacetonate was added to the above system and stirred at 80 ° C for 5 min; the mixture was transferred to a polytetrafluoroethylene autoclave (15 mL) and subjected to a solvothermal reaction at 160 ° C for 12 h; the product was washed with ethanol and centrifuged, repeated 3 times, and dispersed in chloroform to obtain an oil phase Mn / MoO 3-X Nanomaterials.

[0077] Example 3

[0078] A Mn / MoO 3-X The nanomaterial, the preparation method thereof comprises the following steps:

[0079] First, 40 mg of molybdenum acetylacetonate, 200 mg of hexadecylamine and 2.0 mL of oleic acid were added to 8 mL of octadecene, and then uniformly heated at 80 ° C for 10 min; 4 mg of manganese acetylacetonate was added to the above system and stirred at 80 ° C for 5 min; the mixture was transferred to a polytetrafluoroethylene autoclave (15 mL) and subjected to solvothermal reaction at 160 ° C for 12 h; the product was washed with ethanol and centrifuged, repeated 3 times, and dispersed in chloroform to obtain an oil phase Mn / MoO 3-X Nanomaterials.

[0080] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific implementation methods described. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and use the present invention well. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. A Mn / MoO 3-X The method for preparing a nano material is characterized in that: The following steps are involved: (1) Adding a molybdenum source, a fatty amine and a surface stabilizer to a high boiling point solvent, stirring at 60 to 120° C. for 5 to 60 minutes, then adding a manganese source, and continuing to stir at 60 to 120° C. for 1 to 60 minutes; then transferring the mixture to a polytetrafluoroethylene autoclave, performing a solvent thermal reaction at 100 to 280° C. for 6 to 24 hours, washing with an organic polar solvent, centrifuging, and dispersing in an organic non-polar solvent to obtain an oil phase Mn / MoO 3-X Nanomaterial; the mass volume ratio of the high boiling point solvent, the molybdenum source, the fatty amine, the surface stabilizer and the manganese source is 4-12 mL: 4-400 mg: 20-2000 mg: 0.5-10 mL: 4-400 mg; (2) Using a surface modification method, the oil phase Mn / MoO 3-X The nanomaterial and the modified molecule are dissolved in the organic non-polar solvent, stirred at 20-30°C for 0-24h, and then rotary evaporated, and then distilled water is added to disperse to obtain Mn / MoO 3-X Nanomaterials.

2. Mn / MoO according to claim 1 3-X The method for preparing a nano material is characterized in that: In step (1), the high boiling point solvent is one or more of octadecene, benzyl ether, ethylene glycol, N,N-dimethylformamide or dimethyl sulfoxide.

3. Mn / MoO according to claim 1 3-X The method for preparing a nano material is characterized in that: In step (1), the molybdenum source is one or more of molybdenum acetylacetonate, molybdenum acetate, sodium molybdate, carbonyl molybdenum or molybdenum chloride.

4. Mn / MoO according to claim 1 3-X The method for preparing a nano material is characterized in that: In step (1), the surface stabilizer is one or more of oleic acid, oleylamine or oleyl alcohol; and the fatty amine is one or more of hexadecylamine, dodecylamine, tetradecylamine or octadecylamine.

5. Mn / MoO according to claim 1 3-X The method for preparing a nano material is characterized in that: In step (1), the manganese source is one or more of manganese acetylacetonate, manganese oleate, manganese acetate or manganese pentacarbonyl.

6. Mn / MoO according to claim 1 3-X The method for preparing a nano material is characterized in that: The organic non-polar solvent is one or more of dichloromethane, chloroform, n-hexane, cyclohexane, dioxane, o-dichlorobenzene or toluene, and the organic polar solvent is one or more of acetone, ethanol, ethyl acetate, methanol, methyl pyrrolidone, medium-chain alcohol, acetonitrile, N,N-dimethylformamide or dimethyl sulfoxide.

7. Mn / MoO according to claim 1 3-X The method for preparing a nano material is characterized in that: In step (2), the modified molecule is one or more of phospholipid-polyethylene glycol, polyethylene glycol, pluronic, polyvinyl pyrrolidone, polyvinyl alcohol, Tween, vitamin E polyethylene glycol succinate, polylactic acid-co-glycolic acid, polydopamine, meso-dithiosuccinate, cysteine, mercaptosuccinic acid, citric acid or mercaptopropionic acid.

8. A Mn / MoO as claimed in any one of claims 1 to 7 3-X Mn / MoO prepared by the preparation method of nanomaterials 3-X Nanomaterials.

9. Mn / MoO according to claim 8 3-X Application of nanomaterials in the preparation of anti-tumor therapeutic drugs.

10. The Mn / MoO according to claim 9 3-X The use of nanomaterials in the preparation of anti-tumor therapeutic drugs is characterized in that: The Mn / MoO 3-X Nanomaterials generate ROS and damage DNA in the tumor microenvironment while slowly releasing Mn in response 2+ and MoO4 2- , achieving long-term cascade activation of the cGAS / STING pathway and enhancing anti-tumor immune response.

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