FeMoO4 nanomaterial, and preparation method and application thereof

Abstract

The application provides a FeMoO4 nanomaterial, a preparation method and application thereof, and comprises the following steps: S1, adding a molybdenum source, a fatty amine and a surface stabilizer in a high-boiling-point solvent, heating and stirring at 60-120 DEG C for 5-60 min, then adding an iron source, heating and stirring at 60-120 DEG C for 1-60 min; transferring the mixture into a polytetrafluoroethylene autoclave, performing a solvothermal reaction at 100-280 DEG C for 6-24 h, then washing and centrifuging by using an organic polar solvent, and dispersing in an organic nonpolar solvent to obtain an oil-phase FeMoO4 nanomaterial; S2, using a surface modification method, dissolving the oil-phase FeMoO4 nanomaterial and a modification molecule in an organic nonpolar solvent, stirring at 25 DEG C for 0-24 h, then rotary evaporating at 60 DEG C for 1 h, and adding distilled water to obtain the FeMoO4 nanomaterial. 3‑x Under the solvothermal reaction condition, the MoO 3‑x 4 nanomaterial is synthesized and eroded, a large number of defect sites are generated in-situ in the matrix, the defect sites are single-atom anchoring sites, in the system, the anchoring sites are anchored by single-atom Fe, and the single-atom Fe nanomaterial has a metal substitution rate of 20.72 wt%.

Description

Technical Field

[0001] This invention relates to the field of nanomedical materials technology, specifically to a FeMoO4 nanomaterial, its preparation method, and its applications. Background Technology

[0002] Current research has found that major diseases such as malignant tumors, neurological disorders, and cardiovascular diseases are all related to metabolic abnormalities. Taking malignant tumors as an example, tumor development depends on the reprogramming of cellular metabolism. One common characteristic of tumor cell metabolism is its ability to obtain essential nutrients from nutrient-deprived environments and utilize these nutrients to maintain survival and generate new biomass. Furthermore, tumor metabolic reprogramming significantly affects signaling pathways and intercellular interactions, promoting tumor development and progression. However, metabolic regulators have drawbacks such as short in vivo half-life, off-target effects, and interference with normal cellular metabolism.

[0003] Studies have found that xanthine oxidoreductase (XOR) catalyzes the conversion of xanthine to uric acid, and low XOR expression is associated with poor cancer prognosis. Furthermore, xanthine is highly expressed in tumor tissues, and its metabolite uric acid can activate various immune cells. In addition, research has shown that uric acid molecules, by activating the uric acid-NLRP3-IL-1β pathway in macrophages, can stimulate immune cells to secrete pro-inflammatory cytokines such as IL-1β, thereby enhancing the activity of T cells. Although XOR plays an important role in disease metabolism and immune microenvironment remodeling, its application is limited due to the high cost of enzyme preparation and purification, poor operational stability, sensitivity to reaction environments, and difficulty in recycling and reusing the enzyme.

[0004] Single-atom nanomaterials have shown great promise in catalysis and biomedicine due to their high atom utilization and strong metal-matrix interface interactions. The main methods for constructing single-atom nanomaterials include defect engineering strategies, spatial confinement strategies, and designed coordination strategies. Currently, researchers use defect engineering to control the defect sites of metal oxides (such as MoO3, TiO2, and CeO2) and carbon-doped materials. They utilize C, O, S, and metal defects in the support as "traps" to capture mononuclear metal precursors, and then anchor the single atoms through coordination between the metal single atoms and the defect sites, thereby achieving the synthesis of single-atom nanomaterials and increasing the atomic substitution rate. However, under high-temperature synthesis conditions, metal single atoms have high activation energies, and the number of anchoring sites on the matrix is ​​limited. At higher metal dosages, melting and aggregation of single atoms cannot be avoided, resulting in a low atomic substitution rate (not exceeding 5 wt%) for single-atom nanomaterials. Furthermore, natural enzymes that catalyze metabolism, such as XOR, often have complex structures and contain multiple metal catalytic centers, limiting the development of nanomaterials that mimic the activity of metabolic enzymes. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the purpose of this invention is to provide FeMoO4 nanomaterials, their preparation methods, and applications. This invention is based on defect-rich MoO4 nanomaterials. 3-x Achieving heterogeneous single-atom doping in the matrix not only increases the metal substitution rate to 20.72 wt%, but also accurately simulates the Fe in XOR. 2+ and tetrahedron Mo 4+ We constructed an XOR-like FeMoO4 nanomaterial with an active center structure. After intravenous injection, it accumulated in tumor tissue, precisely regulating the xanthine-uric acid metabolic pathway within tumor cells and cascading to activate the uric acid-NLRP3-IL-1β pathway in macrophages, specifically stimulating an anti-tumor immune response and achieving precise metabolic therapy for tumors. This work focuses on the artificial mimicry of natural metabolic enzymes and the regulation of cell interactions, providing nanotechnology and therapeutic strategies for precise metabolic remodeling of major and intractable diseases.

[0006] The first aspect of this invention provides a method for preparing FeMoO4 nanomaterials, comprising the following steps:

[0007] S1. Add a molybdenum source, a fatty amine, and a surface stabilizer to a high-boiling-point solvent, heat and stir at 60-120℃ for 5-60 min, then add an iron source, heat and stir at 60-120℃ for 1-60 min; transfer the mixture to a polytetrafluoroethylene autoclave, and carry out a solvothermal reaction at 100-280℃ for 6-24 h. After washing and centrifugation with an organic polar solvent, disperse the mixture in an organic non-polar solvent to obtain oil-phase FeMoO4 nanomaterials; the mass-volume ratio of the high-boiling-point solvent, the molybdenum source, the fatty amine, the surface stabilizer, and the iron source is 4-12 mL: 4-400 mg: 20-2000 mg: 0.5-10 mL: 4-400 mg;

[0008] S2. Using a surface modification method, the oil-phase FeMoO4 nanomaterial and the modified molecules are dissolved in the organic nonpolar solvent, stirred at 25°C for 0-24 h, rotary evaporated at 60°C for 1 h, and then distilled water is added to obtain the FeMoO4 nanomaterial.

[0009] In one embodiment, in step S1, the high-boiling solvent is one or more of octadecene, dibenzyl ether, ethylene glycol, N,N-dimethylformamide, and dimethyl sulfoxide.

[0010] In one embodiment, in step S1, the molybdenum source is one or more of molybdenum acetylacetonate, molybdenum acetate, sodium molybdate, molybdenum carbonyl, and molybdenum chloride.

[0011] In one embodiment, in step S1, the iron source is one or more of ferric acetylacetone, ferrous acetylacetone, ferrous oleate, ferrous acetate, ferric pentacarbonyl, and ferrous gluconate.

[0012] In one embodiment, in step S1, the surface stabilizer is one or more of oleic acid, oleylamine, and oleyl alcohol; the fatty amine is one or more of hexadecylamine, dodecylamine, tetradecylamine, and octadecylamine.

[0013] In one embodiment, in step S1, the organic nonpolar solvent is one or more of dichloromethane, chloroform, n-hexane, cyclohexane, dioxane, o-dichlorobenzene, and toluene, and the organic polar solvent is one or more of acetone, ethanol, ethyl acetate, methanol, methylpyrrolidone, medium-chain alcohols, acetonitrile, N,N-dimethylformamide, and dimethyl sulfoxide.

[0014] In one embodiment, in step S2, the modifying molecule is one or more selected from phospholipid-polyethylene glycol, polyethylene glycol, Pluronic acid, polyvinylpyrrolidone, polyvinyl alcohol, Tween, vitamin E polyethylene glycol succinate, polylactic acid-glycolic acid copolymer, polydopamine, meso-dimercaptosuccinate sodium, cysteine, mercaptosuccinic acid, citric acid, and mercaptopropionic acid.

[0015] The second aspect of the present invention provides FeMoO4 nanomaterials prepared by the above-described preparation method.

[0016] The third aspect of this invention provides the application of the above-mentioned FeMoO4 nanomaterials in the preparation of products for the treatment of diseases by regulating metabolism and immune microenvironment.

[0017] In one embodiment, the disease metabolic regulation includes diseases such as malignant tumors, neurological and cardiac metabolic disorders.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] 1. In the preparation method of FeMoO4 nanomaterials provided by the present invention, under solvothermal reaction conditions, MoO4 is synthesized and dissolved. 3-x The matrix generates numerous defect sites in situ, enabling efficient adsorption and anchoring of heterogeneous metals. These defect sites serve as anchoring points for single atoms. In this system, these anchoring points are anchored by single-atom Fe, resulting in a metal substitution rate of up to 20.72 wt% for the formed single-atom Fe nanomaterials. Furthermore, the preparation method of FeMoO4 nanomaterials provided by this invention can accurately simulate Fe in XOR. 2+ and tetrahedron Mo 4+ The active site conformation provides a feasible scheme and technical method for mimicking the active site of other natural enzymes.

[0020] 2. In the application of the FeMoO4 nanomaterial provided by this invention, the FeMoO4 nanomaterial can specifically mimic the XOR enzyme to convert xanthine into uric acid. In turn, by activating the uric acid-NLRP3-IL-1β pathway in immune cells and secreting pro-inflammatory cytokines such as IL-1β, it can reshape abnormal pathways related to purines and uric acid, and further reshape the immune microenvironment, thus enabling metabolic therapy for metabolic disorders such as malignant tumors, neurological and cardiac diseases. Attached Figure Description

[0021] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings:

[0022] Figure 1 This is a transmission electron microscope (TEM) image of the FeMoO4 nanomaterial in Example 1 of the present invention;

[0023] Figure 2 This is the elemental analysis (EDS-Mapping) diagram of the FeMoO4 nanomaterial in Example 1 of this invention;

[0024] Figure 3 This is a high-angle annular dark-field scanning transmission electron microscope (HAADF-STEM) image of the FeMoO4 nanomaterial in Example 1 of this invention;

[0025] Figure 4 This is a particle size distribution diagram of the FeMoO4 nanomaterial in Example 2 of the present invention;

[0026] Figure 5 This is a particle size distribution diagram of the FeMoO4 nanomaterial in Example 3 of the present invention;

[0027] Figure 6 This is a particle size distribution diagram of the FeMoO4 nanomaterial in Example 4 of the present invention;

[0028] Figure 7 This is a particle size distribution diagram of the FeMoO4 nanomaterial in Example 5 of the present invention;

[0029] Figure 8 This is an in vitro XOR-like catalytic performance diagram of FeMoO4 nanomaterials in Experimental Example 1 of this invention (xanthine → uric acid).

[0030] Figure 9 This is a graph showing the survival rate of B16 cells with different concentrations of FeMoO4 nanomaterials in Experimental Example 2 of this invention.

[0031] Figure 10 This is a diagram showing the effect of FeMoO4 nanomaterials on macrophage polarization in Experimental Example 3 of this invention;

[0032] Figure 11 This is a graph showing the effect of FeMoO4 nanomaterials on uric acid levels in tumor tissue in Application Example 1 of this invention;

[0033] Figure 12 This is a diagram showing the results of FeMoO4 nanomaterial inhibiting tumor growth in Application Example 2 of this invention;

[0034] Figure 13 This is a diagram showing the effect of FeMoO4 nanomaterials on the polarization of macrophages in tumors in Example 3 of the present invention.

[0035] Figure 14 The FeMoO4 nanomaterials in Example 3 of this invention are used to target CD4 in tumors. + The effect of T cells;

[0036] Figure 15 The FeMoO4 nanomaterials in Example 3 of this invention are used to target CD8 in tumors. + The effect of T cells. Detailed Implementation

[0037] The present invention will now be described in detail with reference to specific embodiments. These embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several changes and improvements without departing from the concept of the present invention. These all fall within the protection scope of the present invention.

[0038] This invention first provides a method for preparing and applying FeMoO4 nanomaterials. FeMoO4 nanomaterials prepared under specific conditions can specifically mimic XOR enzymes, converting xanthine into uric acid. This can reshape abnormal pathways related to purines, uric acid, and other metabolic disorders, and further reshape the immune microenvironment, enabling metabolic therapy for malignant tumors, neurological disorders, cardiac diseases, and other metabolic disorders.

[0039] Next, the preparation method of the above-mentioned FeMoO4 nanomaterials will be described:

[0040] First, a molybdenum source, aliphatic amine, and surface stabilizer are added to a high-boiling-point solvent and heated and stirred at 60-120°C for 5-60 min. Then, an iron source is added, and the mixture is heated and stirred at 60-120°C for 1-60 min. The mixture is then transferred to a polytetrafluoroethylene autoclave and subjected to a solvothermal reaction at 100-280°C for 6-24 h. After washing and centrifugation with an organic polar solvent, the mixture is dispersed in an organic nonpolar solvent to obtain oil-phase FeMoO4 nanomaterials. The mass-volume ratio of the high-boiling-point solvent, the molybdenum source, the aliphatic amine, the surface stabilizer, and the iron source is 4-12 mL: 4-400 mg: 20-2000 mg: 0.5-10 mL: 4-400 mg.

[0041] S2. Using a surface modification method, the oil-phase FeMoO4 nanomaterial and the modified molecules are dissolved in the organic nonpolar solvent, stirred at 25°C for 0-24 h, rotary evaporated at 60°C for 1 h, and then distilled water is added to obtain the FeMoO4 nanomaterial.

[0042] In the above preparation process, the MoO3 matrix was synthesized under atmospheric pressure and heating conditions; under solvothermal reaction conditions, MoO3 matrix rich in defect sites was obtained through aliphatic amine-mediated reduction and in-situ dissolution. 3-x Matrix. MoO 3-x The Mo sites in the matrix can serve as anchoring sites for single atoms, capturing and anchoring metal single atoms to achieve single-atom doping of heterometals; in this system, MoO 3-x Mo vacancies in the matrix are anchored by Fe single atoms. Furthermore, during the anchoring and doping of Fe single atoms using this synthetic strategy, the conformations of Mo and Fe atoms rearrange, resulting in an accurate simulation of Fe in XOR. 2+ and tetrahedron Mo 4+ The crystal structure of the active site conformation provides a feasible scheme and technical method for mimicking the active sites of other natural enzymes.

[0043] In one embodiment, in step S1, the high-boiling solvent is one or more of octadecene, dibenzyl ether, ethylene glycol, N,N-dimethylformamide, and dimethyl sulfoxide.

[0044] In some embodiments, in step S1, the molybdenum source is one or more of molybdenum acetylacetonate, molybdenum acetate, sodium molybdate, molybdenum carbonyl, and molybdenum chloride.

[0045] In some embodiments, in step S1, the iron source is one or more of ferric acetylacetone, ferrous acetylacetone, ferrous oleate, ferrous acetate, ferric pentacarbonyl, and ferrous gluconate.

[0046] In some embodiments, in step S1, the surface stabilizer is one or more of oleic acid, oleylamine, and oleyl alcohol; the fatty amine is one or more of hexadecylamine, dodecylamine, tetradecylamine, and octadecylamine. Oleic acid and fatty amines can slowly generate water molecules in a solvothermal system, which facilitates the in-situ dissolution of the MoO3 matrix and the formation of surface defect vacancies.

[0047] In some embodiments, in step S1, the organic nonpolar solvent is one or more of dichloromethane, trichloromethane, n-hexane, cyclohexane, dioxane, o-dichlorobenzene, and toluene, and the organic polar solvent is one or more of acetone, ethanol, ethyl acetate, methanol, methylpyrrolidone, medium-chain alcohols, acetonitrile, N,N-dimethylformamide, and dimethyl sulfoxide.

[0048] In some embodiments, in step S2, the modifying molecule is one or more of the following: phospholipid-polyethylene glycol, polyethylene glycol, Pluronic acid, polyvinylpyrrolidone, polyvinyl alcohol, Tween, vitamin E polyethylene glycol succinate, polylactic acid-glycolic acid copolymer, polydopamine, meso-dimercaptosuccinate sodium, cysteine, mercaptosuccinic acid, citric acid, and mercaptopropionic acid. The modifying molecule can transform the hydrophobic surface of FeMoO4 nanomaterials into a hydrophilic surface, giving it good colloidal stability.

[0049] The present invention will be further explained in detail below with reference to embodiments:

[0050] Example 1

[0051] This embodiment provides the synthesis and characterization of FeMoO4 nanomaterials.

[0052] 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 the mixture was heated uniformly at 80 °C for 10 min. 50 mg of iron acetylacetonate was added to the above system, and the mixture was stirred at 80 °C for another 5 min. The mixture was then 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 oil-phase FeMoO4 nanomaterials.

[0053] The morphology of the prepared oil-phase FeMoO4 nanomaterials was characterized by TEM, such as... Figure 1 As shown.

[0054] The elemental distribution in FeMoO4 nanomaterials was analyzed using EDS-Mapping, and the results are as follows: Figure 2 As shown.

[0055] The dispersion state of Fe atoms within the FeMoO4 nanomaterial was confirmed using HAADF-STEM, and the results are as follows: Figure 3 As shown.

[0056] Weigh out 20 mg of phospholipid-polyethylene glycol (DSPE-PEG). 2000 2 mg of oil-phase FeMoO4 nanomaterials were dissolved in 5 mL of chloroform. The mixture was stirred at 25 °C for 4 h, rotary evaporated at 60 °C for 1 h, and then dispersed in distilled water to obtain aqueous-phase FeMoO4 nanomaterials.

[0057] Example 2

[0058] First, 40 mg of molybdenum acetylacetonate, 1000 mg of hexadecylamine, and 10.0 mL of oleic acid were added to 8 mL of octadecene, and then the mixture was heated uniformly at 120 °C for 5 min. 400 mg of iron acetylacetonate was added to the above system, and the mixture was stirred at 120 °C for another 5 min. The mixture was then transferred to a polytetrafluoroethylene autoclave (15 mL) and subjected to a solvothermal reaction at 160 °C for 12 h. The product was washed with acetone and centrifuged, and the process was repeated 3 times. The product was then dispersed in dichloromethane to obtain oil-phase FeMoO4 nanomaterials.

[0059] Weigh out 20 mg of phospholipid-polyethylene glycol (DSPE-PEG). 2000 2 mg of oil-phase FeMoO4 nanomaterials were dissolved in 5 mL of chloroform. The mixture was stirred at 25 °C for 4 h, rotary evaporated at 60 °C for 1 h, and then dispersed in distilled water to obtain aqueous-phase FeMoO4 nanomaterials.

[0060] Hydrated particle size analysis was performed on the prepared FeMoO4 nanomaterials, and the results are as follows: Figure 4 As shown.

[0061] Example 3

[0062] 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 the mixture was heated uniformly at 120 °C for 5 min. 4 mg of iron acetylacetonate was added to the above system, and the mixture was stirred at 120 °C for another 5 min. The mixture was then transferred to a polytetrafluoroethylene autoclave (15 mL) and subjected to a solvothermal reaction at 160 °C for 12 h. The product was washed with acetone and centrifuged, repeated 3 times, and dispersed in dichloromethane to obtain oil-phase FeMoO4 nanomaterials.

[0063] Weigh out 20 mg of phospholipid-polyethylene glycol (DSPE-PEG). 2000 2 mg of oil-phase FeMoO4 nanomaterials were dissolved in 5 mL of chloroform. The solution was rotary evaporated at 60 °C for 1 h, and then dispersed in distilled water to obtain aqueous-phase FeMoO4 nanomaterials.

[0064] Hydrated particle size analysis was performed on the prepared FeMoO4 nanomaterials, and the results are as follows: Figure 5 As shown.

[0065] Example 4

[0066] First, 400 mg of molybdenum acetylacetonate, 2000 mg of hexadecylamine, and 10.0 mL of oleic acid were added to 12 mL of octadecene, and then the mixture was heated uniformly at 120 °C for 5 min. 40 mg of iron acetylacetonate was added to the above system, and the mixture was stirred at 120 °C for another 5 min. The mixture was then 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 dichloromethane to obtain oil-phase FeMoO4 nanomaterials.

[0067] Weigh 10 mg of Prönnick F127 and 2 mg of oil-phase FeMoO4 nanomaterials and dissolve them in 2 mL of dichloromethane. Stir at 25 °C for 12 h, rotary evaporate at 60 °C for 1 h, and disperse in distilled water to obtain aqueous-phase FeMoO4 nanomaterials.

[0068] Hydrated particle size analysis was performed on the prepared FeMoO4 nanomaterials, and the results are as follows: Figure 6 As shown.

[0069] Example 5

[0070] First, 4 mg of molybdenum acetylacetonate, 20 mg of hexadecylamine, and 0.5 mL of oleic acid were added to 4 mL of octadecene, and then the mixture was heated uniformly at 100 °C for 5 min. 4 mg of iron acetylacetonate was added to the above system, and the mixture was stirred at 100 °C for another 5 min. The mixture was then transferred to a polytetrafluoroethylene autoclave (15 mL) and subjected to a solvothermal reaction at 280 °C for 6 h. The product was washed with ethanol and centrifuged, repeated 3 times, and dispersed in dichloromethane to obtain oil-phase FeMoO4 nanomaterials.

[0071] Weigh 10 mg of vitamin E polyethylene glycol succinate and 2 mg of oil-phase FeMoO4 nanomaterials and dissolve them in 2 mL of dichloromethane. Stir at 25 °C for 12 h, rotary evaporate at 60 °C for 1 h, and disperse in distilled water to obtain aqueous-phase FeMoO4 nanomaterials.

[0072] Hydrated particle size analysis was performed on the prepared FeMoO4 nanomaterials, and the results are as follows: Figure 7 As shown.

[0073] Experimental Example 1

[0074] This experimental example demonstrates the ability of the FeMoO4 nanomaterials prepared in Example 1 to catalyze the production of uric acid in vitro.

[0075] Phosphate-buffered saline (PBS) and undoped MoO2 were prepared separately. 3-x Nanoparticles (NPs) matrix and FeMoO4 nanomaterials were co-incubated with xanthine at 37°C for 72 h. The uric acid content was then detected using a uric acid detection kit (Wuhan Yilairuit Biotechnology) to evaluate the ability of FeMoO4 nanomaterials to catalyze the production of uric acid.

[0076] Reference Figure 8 As shown, MoO 3-x Nanomaterials do not possess XOR-like properties; however, FeMoO4 nanomaterials mimic the properties of Fe in XOR. 2+ and tetrahedron Mo 4+ The active site conformation enables XOR-like catalytic processes (xanthine → uric acid).

[0077] Experimental Example 2

[0078] This experimental example demonstrates the effect of the FeMoO4 nanomaterials prepared in Example 1 on cytotoxicity.

[0079] B16 cells (6×10) 3 Cells were seeded in 96-well plates and cultured at 37°C for 12 h. After incubation for 24 h with Dulbecco's Modified Eagle Medium (DMEM) containing different concentrations (0, 1.56, 3.13, 6.25, 12.5 μg / mL) of FeMoO4 nanomaterials containing amino acids and glucose, 10 μL of thiazolyl blue (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) stock solution (5 mg / mL) was added to each well, and the cells were incubated at 37°C for 2 h. After removing the MTT medium, the formazan crystal precipitate was dissolved in 100 μL of DMSO, and the absorbance of each well at 570 nm was measured using a microplate reader.

[0080] Reference Figure 9 As shown, FeMoO4 nanomaterials do not exhibit significant cytotoxicity.

[0081] Experimental Example 3

[0082] This experimental example demonstrates the effects of the FeMoO4 nanomaterials prepared in Example 1 on cell interactions and macrophage polarization.

[0083] B16 cells (2 × 10) 4 (Each well) was seeded into a 6-well plate and cultured at 37°C for 12 hours. Cells were then seeded with MoO2-containing reagents. 3-x B16 cells were treated with DMEM containing nanomaterials, uric acid (50 μg / mL), or FeMoO4 nanomaterials (Mo 3.13 μg / mL) for 24 h; Raw264.7 cells (5 × 10⁻⁶) were also treated with DMEM. 4 B16 cells were seeded in 6-well plates and cultured at 37°C for 12 h. They were then treated with interleukin-4 (IL-4) (20 ng / mL) for 12 h, which polarized them to the M2 type. The supernatant of the different groups of B16 cells was added to Raw264.7 cells and incubated for 24 h. The Raw264.7 cells were then collected and stained with CD86 and CD206 antibodies. The ratio of CD86⁺ macrophages (M1 type) and CD206⁺ macrophages (M2 type) was analyzed by flow cytometry.

[0084] Reference Figure 10 As shown, the M1 polarization ratio of macrophages in the FeMoO4 nanomaterial group was significantly stronger than that in the MoO4 group. 3-x Nanomaterials group. The speculated reason is that FeMoO4 nanomaterials induce tumor cells to produce and release uric acid molecules, which effectively induce macrophages to polarize towards the M1 type. This demonstrates that FeMoO4 nanomaterials can influence cell-cell interactions and regulate macrophage polarization within the tumor microenvironment.

[0085] Application Example 1

[0086] This application example demonstrates the use of the FeMoO4 nanomaterials prepared in Example 1 in tumor metabolic remodeling.

[0087] B16 cells (1×10) 6 The suspension was injected into the back of male C57BL / 6 mice. One week later, mice with B16 melanoma were subcutaneously injected with PBS and FeMoO4 nanomaterials (Fe 5 mg / kg) intravenously. The tumor tissues were fixed and sectioned, and spatial metabolomics analysis was used to analyze the regulation of uric acid molecular metabolism in tumor tissues by FeMoO4 nanomaterials.

[0088] Reference Figure 11 As shown, the uric acid level in tumors treated with FeMoO4 nanomaterials was significantly higher than that in the control group. In vivo results indicate that FeMoO4 nanomaterials, with their unique XOR-like enzyme properties, can effectively regulate uric acid metabolism in tumor tissues.

[0089] Application Example 2

[0090] This application example demonstrates the use of the FeMoO4 nanomaterials prepared in Example 1 in their in vivo tumor-suppressing properties.

[0091] B16 cells (1×10) 6 The suspension was injected into the back of male C57BL / 6 mice; one week later, mice bearing B16 melanoma subcutaneously were randomly divided into three groups (n = 5), and intravenously injected with PBS and undoped MoO2 every other day. 3-x After applying nanomaterials, specifically FeMoO4 nanomaterials (Fe 5 mg / kg), the tumor volume was recorded using vernier calipers.

[0092] Reference Figure 12 As shown, FeMoO4 nanomaterials can significantly inhibit tumor growth, and the inhibitory effect is stronger than that of MoO4. 3-x Nanomaterials group.

[0093] Application Example 3

[0094] This application example demonstrates the use of the FeMoO4 nanomaterials prepared in Example 1 in the remodeling of the tumor immune microenvironment.

[0095] B16 cells (1×10) 6 The suspension was injected into the back of male C57BL / 6 mice; one week later, mice with subcutaneous B16 melanoma were intravenously injected with PBS and FeMoO4 nanomaterials (Fe 5 mg / kg); tumor tissue was also collected and treated with 0.6 mg / mL collagenase IV and 0.01 mg / mL deoxyribonuclease I (DNase I). After digestion in culture medium at 37°C for 1 h, the cells were filtered through a 40 μm cell filter and then treated with 36% Percoll layering medium and cleavage red blood cell suspension, respectively. The single-cell suspension was co-incubated with Fc blocking agent, followed by incubation with metal-labeled antibody. Mass cytometry was then used to analyze macrophages and CD8+ in the tumor tissue. + T cells and CD4 + Changes in T cells.

[0096] Reference Figures 13-15 As shown, FeMoO4 nanomaterials can enhance the activity of macrophages and CD8+ cells in tumor tissues. + T cells and CD4 + T cell levels. This may be because FeMoO4 nanomaterials induce tumor cells to produce and release uric acid molecules, thereby activating the uric acid-NLRP3-IL-1β signaling pathway in macrophages, promoting the secretion of inflammatory factors such as IL-1β by macrophages, and polarizing them towards the M1 type. Inflammatory cytokines then promote T cell activation and enhance anti-tumor immune responses.

Claims

1. A preparation method of FeMoO4 nanomaterials, characterized in that, Comprising the following steps: S1, adding a molybdenum source, a fatty amine and a surface stabilizer in a high-boiling solvent, heating and stirring at 60-120℃ for 5-60 min, then adding an iron source, heating and stirring at 60-120℃ for 1-60 min; transferring the mixture to a polytetrafluoroethylene autoclave, carrying out a solvothermal reaction at 100-280℃ for 6-24 h, washing and centrifuging with an organic polar solvent, and dispersing in an organic non-polar solvent to obtain an oil-phase FeMoO4 nanomaterial; the mass-volume ratio of the high-boiling solvent, the molybdenum source, the fatty amine, the surface stabilizer and the iron source is 4-12 mL:4-400 mg:20-2000 mg:0.5-10 mL:4-400 mg; S2, using a surface modification method, dissolving the oil-phase FeMoO4 nanomaterial and a modification molecule in the organic non-polar solvent, stirring at 25℃ for 0-24 h, rotary evaporation at 60℃ for 1 h, and then adding distilled water to obtain a FeMoO4 nanomaterial.

2. The method for preparing FeMoO4 nanomaterials according to claim 1, characterized in that, In the step S1, the high-boiling solvent is one or more of octadecene, dibenzyl ether, ethylene glycol, N,N-dimethylformamide and dimethyl sulfoxide.

3. The method for preparing FeMoO4 nanomaterials according to claim 1, characterized in that, In the step S1, the molybdenum source is one or more of molybdenum acetylacetone, molybdenum acetate, sodium molybdate, carbonyl molybdenum and molybdenum chloride. 4.The method of claim 1, wherein the FeMoO 4 nanomaterial is prepared by the method of claim 1, and wherein the FeMoO 4 nanomaterial has a particle size of 10-100 nm. In the step S1, the iron source is one or more of iron acetylacetone, iron acetylacetone, iron oleate, ferrous acetate, iron pentacarbonyl and ferrous gluconate. 5.The method of claim 1, wherein the FeMoO 4 nanomaterial is prepared by the method of claim 1, and wherein the FeMoO 4 nanomaterial has a particle size of 10-100 nm. In the step S1, the surface stabilizer is one or more of oleic acid, oleylamine and oleyl alcohol; the fatty amine is one or more of hexadecylamine, dodecylamine, tetradecylamine and octadecylamine. 6.The method of claim 1, wherein the FeMoO 4 nanomaterial is prepared by the method of claim 1. In the step S1, the organic non-polar solvent is one or more of dichloromethane, trichloromethane, n-hexane, cyclohexane, dioxane, o-dichlorobenzene and toluene, and the organic polar solvent is one or more of acetone, ethanol, ethyl acetate, methanol, methylpyrrolidone, medium-chain alcohol, acetonitrile, N,N-dimethylformamide and dimethyl sulfoxide.

7. The method for preparing FeMoO4 nanomaterials according to claim 1, characterized in that, In the step S2, the modification molecule is one or more of phospholipid-polyethylene glycol, polyethylene glycol, pluronic, polyvinylpyrrolidone, polyvinyl alcohol, Tween, vitamin E polyethylene glycol succinate, polylactic acid-glycolic acid copolymer, polydopamine, meso-dimercaptosuccinic acid sodium, cysteine, mercaptosuccinic acid, citric acid and mercaptopropionic acid.

8. A FeMoO4 nanomaterial prepared by the preparation method of any one of claims 1-7.

9. Use of the FeMoO4 nanomaterial of claim 8 in the preparation of a product for treating metabolic regulation and immune microenvironment regulation of diseases.

10. Use according to claim 9, characterized in that, The metabolic regulation of diseases includes malignant tumors, neurological and cardiac metabolic abnormalities.

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