A Mn / MoO 3-X Nanomaterials, methods of making and using the same

Abstract

The application belongs to the technical field of nanomedicine, and discloses a Mn / MoO 3‑X Nanometer material, preparation method and application thereof. The material is prepared by the following steps: under normal pressure and at 60-120 DEG C, a MoO3 matrix is prepared by adding a molybdenum source, a fatty amine and a surface stabilizer into a high-boiling-point solvent, then a manganese source is added and stirred and dispersed, and then transferred to an autoclave, and solvent thermal reaction is carried out at 100-280 DEG C for 6-24 hours, and then the oil-phase Mn / MoO 3‑X Nanometer material is obtained after treatment. The oil-phase Mn / MoO 3‑X Nanometer material is surface-modified, and the hydrophobic surface of the nanometer material is changed into a hydrophilic surface, so that the nanometer material has good colloidal stability. The nanometer material is based on a MoO 3‑x Matrix for realizing hetero-manganese monatomic doping, which significantly improves the activation efficiency and persistence of the cGAS / STING pathway, has good enzyme-like catalytic performance and biological safety, can reshape the microenvironment of immunosuppression at the tumor site, and has good specific metal immunotherapy effect.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of nanomedicine, and particularly relates to a Mn / MoO 3-X Nanomaterials, methods of making and using the same. BACKGROUND

[0002] Cancer, as one of the human health killers, has long threatened people's life and health. Immunotherapy has brought revolutionary changes to cancer treatment and provided effective treatment options for different types of tumors. Although immunotherapy has been successful, only a small number of cancer patients show good treatment effects, which is mainly attributed to the strong immune suppression network in the tumor microenvironment. The cyclic-GMP-AMP (cGAMP) synthase (cGAS) can recognize double-stranded DNA of cell membrane, induce the production of cGAMP, activate the stimulator of interferon genes (STING), and thus initiate the anti-tumor immune response. So far, 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, easy production of non-specific and adverse reactions, etc.

[0003] Catalytic therapy, as an emerging strategy for tumor treatment, has attracted extensive attention, which kills tumor cells by converting non-toxic or low-toxic endogenous substances into highly toxic molecules, thereby improving the treatment 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. Nanoenzymes with POD / OXD-like enzyme 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 radical (·OH) etc. ROS induces tumor cell death by damaging intracellular biomolecules (such as proteins, lipids, DNA).

[0004] The manganese element of manganese-based nanoenzymes has multiple different oxidation states, and the manganese atom in low oxidation state can exhibit enzyme-like catalytic activity to produce ROS, while the manganese atom in higher oxidation state can be reduced by glutathione (GSH) and release Mn 2+ . The produced ROS can promote the breakage and leakage of double-stranded DNA, and synergize with the released Mn 2+ to activate the cGAS / STING signaling pathway, realizing the combined catalytic therapy for tumor immunotherapy. However, the reported manganese-based nanoenzymes have unsatisfactory enzyme-like catalytic performance, and the ability to induce ROS production is weak; in addition, membrane transport proteins actively efflux Mn 2+ , leading to the decrease of Mn 2+The concentration is reduced to limit the activation effect of the cGAS / STING pathway, and ultimately leads to the fact that the existing manganese-based nanoscale enzyme cannot achieve good antitumor treatment effect. It can be seen that it is urgent to develop a nano material that can efficiently produce a large amount of ROS, has high tumor cell killing efficiency, has good in vivo biological safety, and can sustainably activate the cGAS / STING pathway to enhance the antitumor effect.

[0005] In recent years, single-atom nanomaterials have shown great potential in catalysis and biomedical fields due to their high atom utilization and strong metal-matrix interface interaction. The construction methods of single-atom nanomaterials mainly include defect engineering strategy, spatial confinement strategy, and design coordination strategy. Studies have found that molybdenum (MoO4 2- ) as a nutritionally essential trace element also has the ability to activate the cGAS-STING pathway, and is more likely to accumulate in cells than Mn 2+ Therefore, based on the team's previous research on molybdenum oxide and defect engineering strategy, the inventors speculate that it is possible to anchor manganese single atoms through the coordination of metal single atoms and defect sites, which can improve ROS production efficiency and make up for the shortcoming of Mn 2+ rapidly excreted, leading to short activation efficiency of cGAS / STING. SUMMARY

[0006] To solve the above problems, the present application provides a Mn / MoO 3-X nanomaterial. The material is based on a MoO 3-x matrix rich in defects to realize hetero-Mn single atom doping, which not only can realize strong peroxidase mimic catalysis to induce severe double-stranded DNA damage in tumors, but also Mn 2+ and MoO4 2- can be slowly released from the Mn / MoO 3-x nanomaterial under the tumor microenvironment to realize long-acting cascade activation of the cGAS / STING pathway, which can be used for tumor-specific catalytic metal immunotherapy and provides a feasible scheme for developing new tumor immunotherapy drugs.

[0007] The technical solutions of the present application are as follows:

[0008] The present application provides a preparation method of a Mn / MoO 3-X nanomaterial, comprising the following steps:

[0009] (1) adding a molybdenum source, a fatty amine and a surface stabilizer into a high-boiling solvent, stirring at 60-120℃ for 5-60min, then adding a manganese source, continuing to stir at 60-120℃ for 1-60min; then transferring the mixture into a polytetrafluoroethylene autoclave, carrying out a solvothermal reaction at 100-280℃ for 6-24h, washing with an organic polar solvent, centrifuging, dispersing in an organic nonpolar solvent to obtain an oil-phase Mn / MoO 3-X nanomaterials; the mass-volume ratio of the high-boiling solvent, the molybdenum source, the fatty amine, the surface stabilizer and the manganese source is 4-12mL:4-400mg:20-2000mg:0.5-10mL:4-400mg;

[0010] (2) using a surface modification method, dissolving the oil-phase Mn / MoO 3-X nanomaterials and the modifying molecules in the organic nonpolar solvent, stirring at 20-30℃ for 0-24h, then rotary evaporation, adding distilled water to disperse to obtain Mn / MoO 3-X nanomaterials.

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

[0012] In an embodiment of the present application, 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 an embodiment of the present application, 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. Among them, oleic acid and fatty amine can slowly generate water molecules in the solvothermal system, which is conducive to accelerating the in-situ dissolution of MoO3 matrix and the formation of surface defect vacancies.

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

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

[0016] In an embodiment of the present application, in step (2), the modifying molecule is preferably one or more of phospholipid-polyethylene glycol, polyethylene glycol, pluronics, polyvinylpyrrolidone, polyvinyl alcohol, Tween, vitamin E polyethylene glycol succinate, polylactic-co-glycolic acid, polydopamine, meso-dimercaptosuccinate sodium, cysteine, mercaptosuccinic acid, citric acid or mercaptopropionic acid. The modifying molecule can convert the hydrophobic surface of the nanomaterial into a hydrophilic surface, so that it has good colloidal stability. 3-X The hydrophobic surface of the nanomaterial is converted into a hydrophilic surface, so that it has good colloidal stability.

[0017] The present application also discloses a Mn / MoO 3-X nanomaterial prepared by the preparation method of the Mn / MoO 3-X nanomaterial.

[0018] The present application also discloses a Mn / MoO 3-X nanomaterial in the preparation of an antitumor therapeutic drug.

[0019] In an embodiment of the present application, the Mn / MoO 3-X nanomaterial slowly releases Mn 2+ and MoO4 2- to activate the cGAS / STING pathway in a long-acting cascade manner.

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

[0021] The present application discloses a Mn / MoO 3-X nanomaterial, which is prepared by first preparing a MoO3 matrix under normal pressure and heating conditions, and then reducing and in-situ eroding the MoO 3-x matrix (defect sites serving as anchoring sites for manganese monomers) through fatty amine mediation under solvent thermal reaction conditions, so as to realize efficient adsorption and anchoring of heterogeneous manganese metals, realize single-atom doping of the heterogeneous metals, and obtain a nanomaterial stably dispersed in water after surface modification. The nanomaterial can realize strong peroxide enzyme simulation catalysis and induce severe double-stranded DNA (dsDNA) damage in tumors; in addition, the Mn / MoO 3-x nanomaterial can release Mn 2+ and MoO4 2- in a high level of GSH environment, improve the sensitivity of cGAS to dsDNA. More importantly, MoO4 2- is released at a relatively slow speed and is easy to accumulate in cells, which makes up for the Mn 2+The continuous outflow causes insufficient cell accumulation, thereby continuously triggering the secretion of type I interferon by cells for 72 hours. Therefore, the long-acting cascade activation of the cGAS / STING pathway by the nanomaterials of the application can be used for tumor-specific catalytic metal immunotherapy, and provides a feasible scheme for the development of new tumor immunotherapy drugs. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 A Mn / MoO 3-X TEM image of the nanomaterials of the application;

[0023] Figure 2 A Mn / MoO 3-X EDS-Mapping image of the nanomaterials of the application;

[0024] Figure 3 A Mn / MoO 3-X HAADF-STEM image of the nanomaterials of the application;

[0025] Figures 4-5 A Mn / MoO 3-X In vitro POD-like enzyme catalytic performance schematic diagram of the nanomaterials of the application;

[0026] Figure 6 A Mn / MoO 3-X Effect of the nanomaterials of the application on B16 cell survival activity schematic diagram;

[0027] Figure 7 A Mn / MoO 3-X Ion (Mn 2+ / MoO4 2- ) release schematic diagram of the nanomaterials of the application in B16 cells;

[0028] Figure 8 A Mn / MoO 3-X Effect of the nanomaterials of the application on B16 cell double-stranded DNA damage schematic diagram;

[0029] Figure 9 A Mn / MoO 3-X Effect of the nanomaterials of the application on B16 cell cGAS / STING pathway activation schematic diagram;

[0030] Figure 10 A Mn / MoO 3-X Inhibition of tumor growth results schematic diagram of the nanomaterials of the application;

[0031] Figure 11 Mn / MoO 3-X Schematic diagram of the results of the tumor metastasis inhibition of the nanomaterials;

[0032] Figure 12 Mn / MoO 3-X Schematic diagram of the influence of the nanomaterials on the intratumoral dendritic cells;

[0033] Figure 13 Mn / MoO 3-X Schematic diagram of the influence of the nanomaterials on the intratumoral CD4 + T cells;

[0034] Figure 14 Mn / MoO 3-X Schematic diagram of the influence of the nanomaterials on the intratumoral CD8 + T cells. DETAILED DESCRIPTION

[0035] In order to make the above objectives, features and advantages of the present application more apparent and comprehensible, the specific embodiments of the present application will be described in detail below with reference to the accompanying drawings.

[0036] In the following description, a large number of specific details are set forth in order to facilitate a thorough understanding of the present application. However, the present application can be implemented in many different ways other than those described herein, and those skilled in the art can make similar generalizations without departing from the scope of the present application, and therefore the present application is not limited to the specific implementations disclosed below.

[0037] Mn / MoO 3-X Nanomaterials, a preparation method thereof comprising the following steps:

[0038] (1) first, add a molybdenum source, a fatty amine and a surface stabilizer into a high-boiling-point solvent, stir at 60-120℃ for 5-60min, then add a manganese source, continue to stir at 60-120℃ for 1-60min; then transfer the mixture into a polytetrafluoroethylene autoclave, carry out a solvothermal reaction at 100-280℃ for 6-24h, then wash with an organic polar solvent, centrifuge, disperse in an organic nonpolar 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 4-12mL:4-400mg:20-2000mg:0.5-10mL:4-400mg;

[0039] (2) using a surface modification method, the oil-phase Mn / MoO 3-XThe nanomaterial and the modifying molecule are dissolved in the organic non-polar solvent, stirred at 20-30 DEG C for 0-24 hours, then spin-evaporated, and then dispersed by adding distilled water to obtain the Mn / MoO 3-X nanomaterial.

[0040] The MoO3 matrix is first synthesized under normal pressure and heating conditions, and then a MoO 3-x matrix rich in defect sites is obtained through reduction and in-situ corrosion mediated by fatty amine under solvent thermal reaction conditions. 3-x The molybdenum sites of the MoO 3-X matrix can serve as anchoring sites of manganese monatomic, and can capture and anchor the metal manganese monatomic to realize hetero-metal manganese monatomic doping.

[0041] Example 1

[0042] The Mn / MoO 3-X nanomaterial is prepared by the following steps:

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

[0044] The oil phase Mn / MoO 3-X nanomaterial is prepared. Figure 1 The morphology of the oil phase Mn / MoO 3-x nanomaterial is characterized by TEM, and as shown in FIG. 1, the Mn / MoO 3-X nanomaterial is clearly shown as a spherical structure with an average size of about 15 nm. Figure 2 The element distribution of the Mn / MoO 3-x nanomaterial is analyzed by EDS-Mapping, and as shown in FIG. 2, the molybdenum, manganese and oxygen elements are uniformly distributed in the Mn / MoO 3-X nanomaterial. Figure 3 The dispersion state of the manganese atoms in the Mn / MoO 3-X nanomaterial is confirmed by HAADF-STEM, and as shown in FIG. 3, the dark spots (yellow circles) are the dispersed manganese atom sites, and the adjacent molybdenum atoms are brighter due to the higher atomic number.

[0045] (2) 20 mg of phospholipid-polyethylene glycol (DSPE-PEG 2000 ) and 2 mg of the 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 using the following method:

[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. Hydrogen peroxide (H2O2) at various concentrations (20, 40, 80, and 100 μM) was then added, along with 20 μL of 3,3′,5,5′-tetramethylbenzidine (TMB) (1 mM). The absorbance at 652 nm was immediately measured using a UV-visible spectrophotometer.

[0049] like Figures 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 the 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 test of nanomaterials is carried out as follows:

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

[0053] As shown in Figure 6 , the Mn / MoO 3-X nanomaterials can effectively induce the production of cell toxicity, and the IC 50 is about 6.8 μg / mL.

[0054] Test Example 3

[0055] This test example 3 provides a test of the ion (Mn 3-X / MoO4 2+ ) release ability of the Mn / MoO 2- nanomaterial prepared in Example 1 in B16 cells, and the test method is as follows:

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

[0057] As shown in Figure 7 , the intracellular Mn 2+ concentration reached a peak at 24 hours and then decreased sharply, and the gradual accumulation of MoO4 2- compensated for the decrease in Mn 2+ activation efficiency, so that the cascade activation of the cGAS / STING pathway was sustained.

[0058] Test Example 4

[0059] This test example 4 provides a test of the effect of the Mn / MoO 3-X nanomaterial prepared in Example 1 on cell double-stranded DNA damage, and the test method is as follows:

[0060] B16 cells (1×10 5The cells were inoculated into confocal culture dishes and cultured in a 37°C cell culture incubator for 24 h. Then, DMEM was replaced with a medium containing Mn / MoO 3-x Nanomaterials (10 μg / mL) were added to fresh DMEM and cultured for 6 hours, 12 hours, and 24 hours, respectively. The cells were washed twice with PBS and fixed with 4% paraformaldehyde solution for 15 minutes. 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 minutes. Then, the cells were rinsed three times with PBS containing Tween 20 and incubated with diluted anti-rabbit Alexa Fluor 488 solution (Wuhan Bost Biotechnology Co., Ltd.) at 37°C for 1 hour. Finally, the cells were observed using a laser confocal microscope.

[0061] like Figure 8 As shown in the immunofluorescence experiment, the Mn / MoO 3-x The nanomaterials group observed the generation of obvious γ-H2AX fluorescence signal, confirming that Mn / MoO 3-x Nanomaterials can significantly induce double-strand 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 hours, cells were centrifuged (1000 rpm, 5 min), harvested, and lysed with cell lysis buffer. Total protein concentration was quantified using a BCA protein assay kit (Thermo Fisher Scientific, USA). Simultaneously, equal amounts of protein (approximately 20 μg) were separated on SDS-polyacrylamide gels and electrotransferred onto polyvinylidene fluoride (PVDF) membranes. The membranes were then 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), stimulator of interferon genes (STING), and phosphorylated-STING (P-STING)) (Cell Signaling Technology, USA), followed by incubation with corresponding secondary antibodies. Protein bands were visualized using enhanced chemiluminescence (ECL) substrate (Thermo Fisher Scientific, USA).

[0065] like Figure 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 100 mm 3 C57BL / 6 mice bearing B16 tumors were randomly divided into four groups (n=5) and intravenously injected with PBS, MoO2, and PBS every 3 days. 3-x NPs (5 mg / kg), Mn 2+ (5 mg / kg) and Mn / MoO 3-x Nanomaterials (5 mg / kg) were administered, and the tumor volume was recorded using a vernier caliper every two days.

[0069] like Figure 10 As shown, the embodiment of the present invention 1Mn / MoO3-X The nanomaterial can significantly inhibit tumor growth, and the inhibition effect is stronger than MoO 3-x The nanomaterial group.

[0070] Application Example 2

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

[0072] B16 cells (2 x 10 5 / mouse) were injected into the left abdominal region of each mouse. After 3 days, B16 tumor cells (2 x 10 5 / mouse) were injected into the right abdominal region of each mouse to establish a bilateral subcutaneous tumor model. When the volume of the left tumor reached about 100 mm 3 , C57BL / 6 mice with B16 tumors were randomly divided into four groups (n = 5), and were intratumorally injected with PBS, MoO 3-x NPs (5 mg / kg), Mn 2+ (5 mg / kg), and Mn / MoO 3-x nanomaterials (5 mg / kg) every 3 days. After the 12th day of treatment, tumor tissues were taken, treated with 0.6 mg / mL collagenase IV and 0.01 mg / mL deoxyribonuclease I (DNase I), filtered using a 40 μm cell filter after digestion in the culture medium at 37 °C for 1 h, and then treated with 36% Percoll gradient solution and red cell lysis solution. The single cell suspension was incubated with an Fc blocking agent, and then the cells isolated from the tumor were stained with corresponding antibodies. Flow cytometry was used to analyze the changes in dendritic cells, CD8 + T cells and CD4 + T cells in the tumor tissues.

[0073] As shown in Figures 11-14 , the Mn / MoO 3-X nanomaterial can promote the maturation of dendritic cells in tumor tissues, is conducive to subsequent antigen presentation, and increases the levels of CD8 + T cells and CD4 + T cells in tumor tissues, ultimately prevents tumor metastasis, successfully inhibits the growth of a remote tumor, and exerts a high-efficiency anti-tumor specific metal immunotherapy effect.

[0074] Example 2

[0075] The Mn / MoO 3-X nanomaterial of this example 2 has a preparation method comprising the following steps:

[0076] First, 40 mg of molybdenum acetylacetone, 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 acetylacetone was added to the above system, and then stirred at 80°C for 5 min; the mixture was transferred to a polytetrafluoroethylene autoclave (15 mL), and then subjected to a solvothermal reaction at 160°C for 12 h; the product was washed and centrifuged with ethanol for 3 times, and then dispersed in chloroform to obtain an oil phase Mn / MoO 3-X Nanomaterials.

[0077] Example 3

[0078] One kind of Mn / MoO 3-X Nanomaterials, a preparation method thereof comprising the following steps:

[0079] First, 40 mg of molybdenum acetylacetone, 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 acetylacetone was added to the above system, and then stirred at 80°C for 5 min; the mixture was transferred to a polytetrafluoroethylene autoclave (15 mL), and then subjected to a solvothermal reaction at 160°C for 12 h; the product was washed and centrifuged with ethanol for 3 times, and then dispersed in chloroform to obtain an oil phase Mn / MoO 3-X Nanomaterials.

[0080] The preferred embodiments of the present application disclosed above are only used to help explain the present application. The preferred embodiments do not describe all the details, and do not limit the present application to the specific embodiments described. Obviously, many modifications and variations can be made according to the content of the present application. The present application selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present application, so that those skilled in the art can well understand and utilize the present application. The present application is limited only by the claims and their full scope and equivalents.

Claims

1. A Mn / MoO 3-X The method for preparing a nanomaterial 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 60-120°C for 5-60 minutes, then adding a manganese source, and continuing to stir at 60-120°C for 1-60 minutes; then transferring the mixture to a polytetrafluoroethylene autoclave, performing a solvent thermal reaction at 100-280°C for 6-24 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 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 nanomaterial 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 nanomaterial is characterized in that: In step (1), the molybdenum source is one or more of molybdenum acetylacetonate, molybdenum acetate, sodium molybdate, molybdenum carbonyl or molybdenum chloride.

4. Mn / MoO according to claim 1 3-X The method for preparing a nanomaterial 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 nanomaterial 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 nanomaterial is characterized in that: The organic non-polar solvent is one or more of dichloromethane, chloroform, n-hexane, cyclohexane, dioxane, o-dichlorobenzene or toluene; 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 nanomaterial 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-dimercaptosuccinate, cysteine, mercaptosuccinic acid, citric acid or mercaptopropionic acid.

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

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

10. The Mn / MoO according to claim 9 3-X The application 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 responses.

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