Bimetal nanoparticles as well as preparation method and application thereof

Abstract

The invention relates to the technical field of nanomedicine and functional material crossing, and discloses a bimetallic nanoparticle and a preparation method and application thereof.The bimetallic nanoparticle comprises metal A and metal B and is coated with protein polypeptide substances, and the preparation method comprises the steps that the protein polypeptide substances are evenly suspended in water, a metal solution and a carbonate solution are added, and then the pH of the solution is adjusted to be alkaline; the preparation method comprises the following steps: adding a proper reaction temperature, a proper time and a reaction system, dialyzing, freezing and drying to obtain bimetallic nanoparticles, using an ultrafiltration centrifuge tube to obtain the bimetallic nano quantum dots, and applying the bimetallic nanoparticles to regeneration and repair of tumor and tissue damage, osteolysis related to prosthesis, and treatment of cardiovascular and metabolic diseases. The bimetallic nanoparticles have the advantages of good repeatability, uniform particle size distribution, controllable structure, multiple functions and excellent biocompatibility, solve the problem of poor targeting property of existing drugs, are suitable for the fields of precise treatment and drug delivery, and also have application prospects in many fields of daily chemicals, batteries, environmental protection and the like.

Description

Technical Field

[0001] This invention relates to the field of interdisciplinary technology of nanomedicine and functional materials, specifically a bimetallic nanoparticle, its preparation method, and its application. Background Technology

[0002] With the rapid development of nanotechnology in the biomedical field, nanoparticles have shown significant therapeutic potential as drug delivery systems. However, traditional single-metal nanoparticles still have significant shortcomings in terms of loading efficiency, controlled release capability, bioactivity, and multifunctional synergy, which limits their application in the treatment of complex diseases.

[0003] In recent years, bimetallic nanomaterials have attracted widespread attention in fields such as catalysis, imaging, antibacterial agents, and targeted therapy due to their unique physicochemical properties and synergistic effects with metals. Compared with single-metal materials, bimetallic nanoparticles have significant advantages in terms of stability, tunability, and functional integration, making them particularly suitable for synergistic treatment of multi-mechanism diseases such as tumors and inflammatory bone diseases.

[0004] However, current methods for preparing bimetallic nanoparticles are complex and costly. Therefore, there is an urgent need to develop bimetallic nanomaterials with controllable structure, tunable function, and good biocompatibility, along with efficient synthesis strategies. Among these methods, using proteins as templates or coating materials to prepare nanoparticles can not only improve their biocompatibility and stability but also endow them with good targeting and modifiability.

[0005] Existing methods for preparing metal nanomaterials include chemical reduction, hydrothermal synthesis, and biosynthesis, but most of these methods have the following technical problems: (1) The preparation process is complex or the conditions are harsh, such as high temperature and high pressure, the use of organic solvents or highly toxic reducing agents, which is not conducive to subsequent biomedical applications; (2) The nanoparticle size is poorly controlled, resulting in large fluctuations in material properties and affecting actual functions; (3) The materials have poor dispersibility and biocompatibility, which can easily trigger immune reactions or aggregate inactivation in vivo; (4) It is difficult to achieve a stable and synergistic distribution of the bimetallic system, which affects its multifunctional combined therapeutic effect.

[0006] Document CN 114939165 A discloses reversible multidrug resistance bimetallic nanoparticles, their preparation method, and applications. The method involves adding a carboxyl-containing acid solution to a solution of bimetallic nanoparticles, adding an anticancer drug, and stirring to obtain anticancer drug@bimetallic-COOH nanoparticles. A solution of bovine serum albumin is reacted with a magnesium salt, followed by the addition of sodium hydroxide solution to continue the reaction, yielding a BSA-Mg complex. The BSA-Mg complex is then loaded onto the anticancer drug@bimetallic-COOH nanoparticles. This method cannot be used as a drug carrier.

[0007] Therefore, a new technical solution is needed to solve the above-mentioned technical problems. Summary of the Invention

[0008] To address the aforementioned issues, this invention discloses a bimetallic nanoparticle, its preparation method, and its applications. This invention provides a bimetallic nanomaterial with multifunctional synergistic therapeutic capabilities and its preparation method, solving problems such as poor biocompatibility, incomplete functionality, and complex processes in existing technologies. It also demonstrates the potential for application in complex disease scenarios such as targeted drug delivery, immune regulation, prosthesis-related osteolysis, and tumor microenvironment intervention.

[0009] The technical solution of the present invention is as follows: a bimetallic nanoparticle, wherein the bimetallic nanoparticle includes metals A and B selected from any two of copper, silver, manganese, iron, magnesium, aluminum, and calcium, the bimetallic nanoparticle is coated with a protein polypeptide substance, and contains a first metal element A and a second metal element B, wherein metals A and B are selected from any two of copper, silver, manganese, iron, magnesium, aluminum, and calcium, and the molar ratio of metal elements A and B is 1:1 to 5:1.

[0010] A method for preparing bimetallic nanoparticles, wherein the bimetallic nanoparticles are obtained by a biomimetic mineralization method, comprising the following steps: Step 1: Evenly suspend the protein and polypeptide substances in water, add metal solution and carbonate solution, and then adjust the pH of the solution to alkaline; Step 2: After dialysis and freeze-drying at a suitable reaction temperature, time, and reaction system, bimetallic nanoparticles are obtained, and bimetallic quantum dots are further obtained using ultrafiltration centrifuge tubes.

[0011] By adopting the above technical solution and preparing the bimetallic nanoparticles through biomimetic mineralization, they exhibit good biocompatibility and controllable particle size. They can release metal ions in the inflammatory or tumor microenvironment to regulate the immune microenvironment or induce cell death. The obtained bimetallic nanoparticles have a controllable particle size in the range of 5 nanometers to 100 nanometers and good dispersibility. They can be used in daily chemical, battery, environmental protection, and pharmaceutical fields, and can also be loaded with drugs such as anti-tumor drugs doxorubicin or 5-fluorouracil.

[0012] Preferably, the protein-peptide substance in step 1 is one or more of bovine serum albumin, human serum albumin, collagen, gelatin, angiogenesis factor, epidermal growth factor, auxin, insulin, interferon, subunit protein-peptide vaccine, and antibody, and its concentration in the reaction solution is 0.1 mg / mL to 100 mg / mL.

[0013] Preferably, in step 1, the concentration of the two metal salt solutions is 0.01 mg / mL to 100 mg / mL, the carbonate ion is derived from sodium carbonate, potassium carbonate, sodium bicarbonate or potassium bicarbonate, and the concentration is 0.01 mg / mL to 100 mg / mL, with a slight excess of carbonate solution.

[0014] Preferably, in step 1, the pH of the reaction is not less than 10, and the pH is adjusted by sodium hydroxide or potassium hydroxide.

[0015] Preferably, in step 2, the pH of the reaction system is adjusted to 10-13, sodium hydroxide or potassium hydroxide is used, the reaction temperature is 30°C-100°C, the reaction time is 0.5 h-6 h, and the stirring speed is 300 rpm-1000 rpm.

[0016] Preferably, in step 2, the reaction product is dialyzed with ultrapure water, the molecular weight cutoff of the dialysis bag is 1.0 kDa to 100 kDa, the dialysis temperature is 4°C to 25°C, the dialysis time is 8 h to 72 h, and the pore size of the ultrafiltration tube is 1.0 kDa to 100.0 kDa.

[0017] Preferably, in step 2, the average diameter of the particles is 5 nm to 100 nm, the morphology is near-spherical, the particle size distribution is narrow, and the PDI is < 0.2, so that bimetallic quantum nanodots with a particle size < 10 nm can be further prepared.

[0018] By adopting the above technical solution, bimetallic nanoparticles have good dispersibility and can be used in daily chemical, battery, environmental protection, and pharmaceutical fields. They can also be loaded with drugs such as anti-tumor drugs doxorubicin or 5-fluorouracil.

[0019] Applications of bimetallic nanoparticles in the treatment of tumors, tissue damage regeneration and repair, prosthesis-related osteolysis, cardiovascular and metabolic diseases.

[0020] By employing the above-mentioned technical solutions, bimetallic nanoparticles can be used in prosthetic-associated osteolysis (PAO) and tumor therapy to accelerate the release of drugs or metal ions under pH ≤ 6.5 or reducing (GSH) conditions. This allows for tumor microenvironment-responsive therapy or periprosthetic microenvironment-responsive therapy. In PAO, they can induce macrophage polarization, scavenge reactive oxygen species (ROS), and inhibit osteoclast differentiation pathways, thereby blocking the vicious cycle of inflammation-bone destruction. In tumor therapy, they can achieve controlled drug release in response to acidic microenvironments and high glutathione (GSH) levels, and synergistically activate ROS generation to induce tumor cell death.

[0021] The advantages of this invention are: 1. This invention can prepare bimetallic nanoparticles by mixing and reacting a uniform albumin solution, metal solution and carbonate solution through a simple biomimetic mineralization method. It has the advantages of mild reaction conditions, simple process flow, cheap and readily available raw materials, short preparation cycle, good biosafety, high repeatability, excellent physicochemical properties and good prospects for large-scale production.

[0022] 2. The bimetallic nanoparticles of this invention can serve as delivery carriers for various drugs (such as the antitumor drugs doxorubicin or 5-fluorouracil), and achieve drug release through disease microenvironment response mechanisms (such as acidic environment, GSH reduction environment) or inflammatory response mechanisms. Animal experiments have verified that the particles have shown good therapeutic effects and biosafety in prosthetic-associated osteolysis (PAO) and solid tumor models.

[0023] 3. The present invention has the advantages of mild reaction conditions, simple process flow, short preparation cycle and good prospects for large-scale production. The bimetallic nanoparticles prepared have the advantages of good reproducibility, uniform particle size distribution, controllable structure, multiple functions and excellent biocompatibility. They can be used to solve the problems of poor targeting and large toxic side effects of existing drug systems. They are suitable for precision treatment and drug delivery, and also have application prospects in many fields such as daily chemicals, batteries and environmental protection. Attached Figure Description

[0024] Figure 1 These are transmission electron micrographs of CuAg-BSA (a) and CuAg-BSA / 5-FU (b); Figure 2 This represents the particle size change of CuAg-BSA over 14 days; Figure 3 These are laser confocal images of reactive oxygen species in 4T1 cells after CuAg-BSA stimulation; Figure 4 Images of H&E staining of the heart, liver, spleen, lungs, and kidneys of different groups of mice (scale bar 100 μm). Figure 5 These are curves comparing tumor volumes in mice from different drug administration groups; Figure 6 These are TUNEL-stained images of tumors in mice from different drug administration groups; Figure 7 The results of CCK8 in 4T1 cells were obtained 24 hours after different groups were stimulated. Figure 8 This is the XRD pattern of CMC; Figure 9It is a cluster heatmap of representative macrophage immunophenotypes, osteoclast differentiation, and aerobic respiration-related genes; Figure 10 It is a transcriptomic analysis of CMC regulation of macrophage immune and metabolic phenotypes; Figure 11 This is an in vitro experimental verification that CMC induces copper death in osteoclasts in response to an acidic environment at the end of osteoclast differentiation; Figure 12 CMC reduces bone loss in a mouse craniocerebral osteolysis model induced by titanium particles.

[0025] The attached image contains color. Detailed Implementation

[0026] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Example

[0027] Step 1: Weigh 60.0 mg BSA and dissolve it in 9 mL of deionized water. Stir magnetically at 550 rpm for 30 min. Slowly add 1 mL of 0.1 mol / L Na2CO3 solution. After the addition is complete, stir at 550 rpm for 30 min at 60℃. Then, slowly add 1.5 mL of 0.04 mol / L CuCl2 solution, 1.0 mL of 0.04 mol / L AgNO3 solution, and finally slowly add 800 μL of 0.1 mol / L NaOH solution. After the solution gradually becomes clear, wrap it in aluminum foil to protect it from light, and stir at 550 rpm for 2 h at 60℃ to obtain CuAg-BSA nanoparticles. Step 2: Dialyze at room temperature in the dark for 48 hours, changing the water every 8 hours to obtain pure CuAg-BSA nanoparticles.

[0028] from Figure 1 As can be seen from 'a', the prepared CuAg-BSA nanoparticles are regular spheres with a small particle size of approximately 20-30 nm. No aggregation is observed in the CuAg-BSA nanoparticles over a wider viewing angle, indicating good dispersibility. Furthermore, the particle shape and size are essentially consistent, demonstrating good uniformity.

[0029] from Figure 2 It can be seen that the synthesized nanoparticles have excellent stability.

[0030] Using 5-fluorouracil as a model drug, the effect of CuAg-BSA nanoparticles on killing cancer cells was evaluated. 5-FU was loaded using an adsorption method. A 5 mg / mL 5-FU solution was prepared with deionized water. 6 mL of the prepared CuAg-BSA solution and 8 mL of the 5-FU solution were slowly added sequentially to a round-bottom flask and thoroughly mixed. The mixture was stirred at 750 rpm for 3 h at room temperature to obtain a CuAg-BSA / 5-FU solution. Subsequently, the ability of CuAg-BSA and CuAg-BSA / 5-FU to activate reactive oxygen species in mouse breast cancer 4T1 cells was evaluated using laser confocal microscopy. 4T1 cells were loaded at 5 × 10⁻⁶... 4 After seeding nanoparticles at a density of 1 nanoparticle per well on a 24-well plate for 24 hours, nanoparticles were added. Reactive oxygen species (ROS) staining was performed 24 hours later, and the fluorescence intensity of each group was observed and photographed using a laser confocal scanning microscope.

[0031] from Figure 1 As can be seen from b, the morphology of CuAg-BSA / 5-FU changes to an elongated shape, and the particle size is significantly larger than that of CuAg-BSA nanomaterials, mainly distributed between 50 and 100 nm, indicating relatively good material dispersion.

[0032] from Figure 3 It can be seen that CuAg-BSA and CuAg-BSA / 5-FU have strong reactive oxygen species activation capabilities, demonstrating their potential application in the field of tumor treatment. Specifically, they can be used in the preparation of anti-tumor drugs.

[0033] from Figure 4 It can be seen that CuAg-BSA and CuAg-BSA / 5-FU have good biocompatibility and tissue compatibility.

[0034] from Figure 5 It can be seen that the tumor volume of mice in the Control group was generally larger at the end, while the tumor volume of mice in the material group and the drug-loaded group showed a relatively slow and similar growth trend with no obvious individual differences, demonstrating that CuAg-BSA and CuAg-BSA / 5-FU have relatively stable and uniform tumor-suppressing effects.

[0035] from Figure 6 It can be seen that the drug-loaded nanomaterials have the strongest tumor cell killing effect. Example

[0036] Step 1: Slowly add 1 mL of 0.1 mol / L Na₂CO₃ solution dropwise to 7 mL of 10 mg / mL BSA solution. After the addition is complete, stir at 500 rpm for 30 min at 55℃. Then, slowly add 1.0 mL of 0.06 mol / L CuCl₂ solution and 1.0 mL of 0.03 mol / L MnCl₂ solution dropwise in sequence, adjust the pH to 10, and stir at 500 rpm for 2 h at 55℃ to obtain CuMn-BSA nanoparticles. Step 2: Dialyze at room temperature for 48 hours, changing the water every 8 hours to obtain pure CuMn-BSA (CMB) nanoparticles.

[0037] Using doxorubicin as a model drug, the effect of CuMn-BSA nanoparticles on killing cancer cells was evaluated. Doxorubicin was loaded onto the nanoparticles using the same method to obtain CuMn-BSA / DOX (CMD). The Cell Counting Kit-8 (CCK-8) assay was used to evaluate the killing effects of CuMn-BSA, DOX, and CuMn-BSA / DOX on 4T1 cells after 24 hours.

[0038] from Figure 7 It can be seen that cell viability is significantly correlated with CMD concentration. At the same dose, CMD exhibits superior anticancer activity compared to CMB and DOX. Example

[0039] Step 1: Slowly add 1 mL of 0.1 mol / L Na₂CO₃ solution dropwise to 7 mL of 10 mg / mL BSA solution. After the addition is complete, stir at 500 rpm for 30 min at 55℃. Then, slowly add 1.0 mL of 0.04 mol / L CuCl₂ solution and 1.0 mL of 0.04 mol / L MnCl₂ solution dropwise, adjusting the pH to 10. Stir at 500 rpm for 2 h at 55℃ to obtain CuMn-BSA nanoparticles. Step 2: Dialyze at room temperature for 48 hours, changing the water every 8 hours to obtain pure CuMn-BSA (CMC) nanoparticles. The crystal structure and composition of CMC were studied using X-ray diffraction (XRD). from Figure 8 As can be seen, the peaks are broad and diffuse, with the peak centers located around 15° and 23°. There is no obvious crystal reflection, indicating that the material is mainly amorphous or has low crystallinity. The characteristic peaks of CuCO3

[011] ,

[202] ,

[131] and MnCO3

[104] ,

[113] can be seen in the figure, but they are all shifted, indicating that Cu-Mn co-doped carbonate crystals have been formed. The absence of obvious diffraction peaks indicates that CMC exists in the BSA matrix in the form of nanoscale domains. Example

[0040] Experimental data and mechanism validation of bimetallic nanoparticles (CMCs) in the treatment of PAO. LPS-stimulated macrophages exhibit significant anti-inflammatory effects, mainly through the scavenging of ROS (reactive oxygen species). Figure 9 A) and reverse M1 polarization (downregulate iNOS, upregulate CD206), while promoting M2 polarization ( Figure 9 F). RT-qPCR confirmed that it inhibited the expression of M1 polarization genes (Cxcl10, Tnf, Il1b, Nos2, and Il6) and promoted the expression of M2 polarization genes (Arg1, Clec10a, Cxcr4, and Mrc1). Figure 9 E), ELISA confirmed that CMC inhibited the expression of macrophage inflammatory cytokines (TNF-α, IL-6, IL-1β) ( Figure 9 JL); Transcriptome sequencing was performed on macrophages treated with the Control group, LPS, and LPS+CMC group to further investigate the effect of CMC on subsequent osteoclast formation. Figure 10 It can be seen that CMC reverses LPS-induced glycolysis and enhances the TCA cycle (upregulating genes such as Mdh1 and Fh1). Figure 10 C, D). Enrichment of the KEGG pathway indicates that CMC activates oxidative phosphorylation (C, D). Figure 10 E), inhibiting NF-κB and osteoclast differentiation pathways ( Figure 10 F) provides a basis for blocking the vicious cycle of inflammation-osteoclasts.

[0041] During the later stages of osteoclast differentiation (acidic environment, pH ≈ 4.5) Figure 11 A) CMC releases Cu² + It induces oligomerization of mitochondrial liponylated protein (DLAT), leading to TCA cycle collapse. Western blot showed increased DLAT oligomers and depletion of FDX1 / LIAS. Figure 11 C), JC-1 staining confirmed the collapse of mitochondrial membrane potential (C), Figure 11 HJ).

[0042] In a mouse skull model induced by titanium particles, local injection of CMC (20 μg / mL) significantly inhibited osteolysis. Micro-CT scans showed increased bone volume fraction (BV / TV) and trabecular bone thickness (Tb.Th), demonstrating superior efficacy compared to ALN (Alpha-Lactobacillus). Figure 12 BF), tissue section staining demonstrated that the addition of CMC reduced osteoclast numbers, suppressed local inflammatory responses, and increased DLAT oligomerization and FDX1 consumption ( Figure 12 (GN), demonstrating its application potential in PAO.

[0043] This embodiment uses BSA as a template to construct Cu-Mn co-doped carbonate nanoparticles (CMCs). The preparation process is mild and simple, and the resulting material exhibits a low-crystallinity nanostructure, which facilitates ion release and cellular uptake. CMCs demonstrate good anti-inflammatory and immunomodulatory capabilities in inflammatory environments, scavenging ROS, inhibiting M1 polarization, promoting M2 differentiation, and downregulating the expression of multiple inflammation-related genes. Transcriptomic and metabolic pathway analyses show that they block inflammation-driven osteoclast differentiation by enhancing the TCA cycle, inhibiting glycolysis, and the NF-κB signaling pathway. In an acidic microenvironment, CMCs release Cu... 2+ It induces mitochondrial DLAT oligomerization, further disrupting osteoclast function. Animal experiments have demonstrated that it significantly inhibits bone resorption in a titanium particle-induced prosthesis-related osteolysis model, with effects superior to conventional therapeutic drugs, exhibiting good biocompatibility and clinical translational potential. Example

[0044] Step 1: Slowly add 1 mL of 0.1 mol / L Na₂CO₃ solution dropwise to 7 mL of 10 mg / mL BSA solution. After the addition is complete, stir at 500 rpm for 30 min at 55℃. Then, slowly add 1.0 mL of 0.1 mol / L CaCl₂ solution and 1.0 mL of 0.1 mol / L CuCl₂ solution dropwise in sequence, adjust the pH to 10, and stir at 500 rpm for 2 h at 55℃ to obtain CaCu-BSA nanoparticles. Step 2: Dialyze at room temperature for 48 hours, changing the water every 8 hours to obtain pure CaCu-BSA nanoparticles. Example

[0045] Step 1: Slowly add 1 mL of 0.1 mol / L Na₂CO₃ solution dropwise to 7 mL of 10 mg / mL BSA solution. After the addition is complete, stir at 500 rpm for 30 min at 65℃. Then, slowly add 1.0 mL of 0.1 mol / L CuCl₂ solution and 1.0 mL of 0.1 mol / L FeCl₃ solution dropwise in sequence, adjust the pH to 11, and stir at 500 rpm for 3 h at 65℃ to obtain CuFe-BSA nanoparticles. Step 2: Dialyze at room temperature for 12 hours, changing the water every 4 hours to obtain pure CuFe-BSA nanoparticles. Then, centrifuge at 10,000 rpm for 10 minutes using an ultrafiltration centrifuge tube to obtain nano-quantum dots. Example

[0046] Step 1: Slowly add 1 mL of 0.05 mol / L Na₂CO₃ solution dropwise to 10 mL of 10 mg / mL BSA solution. After the addition is complete, stir at 450 rpm for 30 min at 50 °C. Then, slowly add 1.0 mL of 0.1 mol / L CuCl₂ solution and 1.0 mL of 0.05 mol / L AlCl₃ solution dropwise in sequence, adjust the pH to 12, and stir at 450 rpm for 3 h at 50 °C to obtain CuAl-BSA nanoparticles. Step 2: Dialyze at room temperature for 12 hours, changing the water every 4 hours to obtain pure CuAl-BSA nanoparticles. Example

[0047] Step 1: Slowly add 2 mL of 0.1 mol / L Na₂CO₃ solution dropwise to 10 mL of 5 mg / mL collagen solution. After the addition is complete, stir at 350 rpm for 30 min at 40℃. Then, slowly add 1.0 mL of 0.1 mol / L FeCl₃ solution and 1.0 mL of 0.1 mol / L AlCl₃ solution dropwise in sequence, adjust the pH to 11, and stir at 350 rpm for 3 h at 40℃ to obtain FeAl-Collagen nanoparticles. Step 2: Dialyze at room temperature for 24 hours, changing the water every 4 hours to obtain pure FeAl-Collagen nanoparticles.

[0048] This example further verifies the feasibility of using collagen instead of BSA as the template protein, through the complexation of Fe... 3+ With Al 3+ Ions were used to construct FeAl-Collagen nanoparticles, expanding the diversity of CMC-type materials in protein templates and metal systems. This method not only preserved the mineralization process under mild conditions, ensuring the bioactivity of collagen, but also endowed the nanoparticles with stronger tissue affinity and cartilage repair potential, providing a new approach for constructing integrated nanosystems with tissue induction and therapeutic functions.

[0049] Those skilled in the art should understand that the embodiments of the present invention described above and shown in the accompanying drawings are merely examples and do not limit the present invention; the objectives of the present invention have been fully and effectively achieved. The functions and structural principles of the present invention have been demonstrated and explained in the embodiments, and any modifications or variations of the embodiments of the present invention may be made without departing from the stated principles.

Claims

1. A bimetallic nanoparticle, characterized in that, The bimetallic nanoparticles include metals A and B selected from any two of copper, silver, manganese, iron, magnesium, aluminum, and calcium. The bimetallic nanoparticles are coated with protein-peptide substances and contain a first metal element A and a second metal element B. Metals A and B are selected from any two of copper, silver, manganese, iron, magnesium, aluminum, and calcium. The molar ratio of metal elements A and B is 1:1 to 5:

1.

2. The method for preparing bimetallic nanoparticles according to claim 1, characterized in that, The bimetallic nanoparticles were prepared by a biomimetic mineralization method, including the following steps: Step 1: Evenly suspend the protein and polypeptide substances in water, add metal solution and carbonate solution, and then adjust the pH of the solution to alkaline; Step 2: After dialyzing and freeze-drying at a suitable reaction temperature, time and reaction system, bimetallic nanoparticles are obtained, and bimetallic quantum dots are obtained by further using ultrafiltration centrifuge tubes.

3. The method for preparing bimetallic nanoparticles according to claim 2, characterized in that, The protein and polypeptide substances mentioned in step 1 are one or more of bovine serum albumin, human serum albumin, collagen, gelatin, angiogenesis factor, epidermal growth factor, auxin, insulin, interferon, subunit protein polypeptide vaccine, and antibody, and their concentration in the reaction solution is 0.1 mg / mL to 100 mg / mL.

4. A method for preparing bimetallic nanoparticles according to claim 2, characterized in that, In step 1, the concentration of the two metal salt solutions in the synthesis is 0.01 mg / mL to 100 mg / mL. The carbonate ion is derived from sodium carbonate, potassium carbonate, sodium bicarbonate, or potassium bicarbonate, and the concentration is 0.01 mg / mL to 100 mg / mL. The carbonate solution is slightly in excess.

5. A method for preparing bimetallic nanoparticles according to claim 2, characterized in that, In step 1, the reaction pH is not less than 10, and the pH is adjusted using sodium hydroxide or potassium hydroxide.

6. A method for preparing bimetallic nanoparticles according to claim 2, characterized in that, In step 2, the pH of the reaction system is adjusted to 10-13, sodium hydroxide or potassium hydroxide is used, the reaction temperature is 30°C-100°C, the reaction time is 0.5 h-6 h, and the stirring speed is 300 rpm-1000 rpm.

7. A method for preparing bimetallic nanoparticles according to claim 2, characterized in that, In step 2, the reaction product is dialyzed with ultrapure water. The molecular weight cutoff of the dialysis bag is 1.0 kDa to 100 kDa, the dialysis temperature is 4°C to 25°C, the dialysis time is 8 h to 72 h, and the pore size of the ultrafiltration tube is 1.0 kDa to 100.0 kDa.

8. A method for preparing bimetallic nanoparticles according to claim 2, characterized in that, In step 2, the average diameter of the particles is 5 nm to 100 nm, the morphology is near-spherical, and there is a narrow particle size distribution with PDI < 0.

2. Bimetallic quantum nanodots with a particle size < 10 nm are further prepared.

9. Applications of bimetallic nanoparticles in the treatment of tumors, tissue damage regeneration and repair, prosthesis-related osteolysis, cardiovascular and metabolic diseases.

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