Preparation method and anti-tumor application of silver / manganese dioxide hybrid nanoparticles

Abstract

The invention provides a silver / manganese dioxide hybrid nanoparticle (Ag (at) MnO-sis-c-L) as well as a preparation method and application thereof. The phospholipid layer modified cRGD targeting molecule can specifically recognize high-expression integrin alpha v beta 3 on the surface of a tumor cell, enrichment of a contrast agent in a tumor tissue is achieved, and the imaging resolution and focus positioning accuracy are improved. Wherein a phospholipid material (HSPC) and PEGylated lipid (DMG-PEG2000 and DSPE-PEG2000-cRGD) reduce immunogenicity and toxic and side effects and ensure in-vivo circulation stability, and a phospholipid layer wraps to improve particle dispersity and ensure in-vivo stability. The inner core is further loaded with siRNA, and the drug delivery function is endowed with the targeting modification and phospholipid wrapping characteristics of the shell, so that diagnosis and treatment integration is realized, and a new platform is provided for precise medical treatment. According to the Ag-coated MnO-sis-c-L contrast agent prepared by the invention, a silver / manganese dioxide coprecipitation core and a phospholipid wrapping structure are creatively combined, and the contrast agent shows remarkable application potential in the aspect of tumor targeted therapy.

Description

Technical Field

[0001] The present invention belongs to the field of medicine preparation, and particularly relates to a preparation method and anti-tumor application of silver / manganese dioxide hybrid nanoparticles. Background Art

[0002] Malignant tumors are a major threat to human health. Existing chemotherapeutic drugs suffer from poor targeting, severe side effects, and tumor resistance, necessitating the development of new, highly effective and low-toxic anti-tumor strategies. Advances in nanotechnology have provided new avenues for precision cancer treatment. Synergistic systems of metal-based nanomaterials and functional oxides have attracted significant attention due to their multiple therapeutic benefits. Silver (Ag) nanoparticles possess a unique surface plasmon resonance effect and broad-spectrum bioactivity, but their single application suffers from limitations such as insufficient stability and limited responsiveness to the tumor microenvironment. Manganese dioxide (MnO2), a typical redox-responsive material, can specifically degrade high concentrations of hydrogen peroxide (H2O2) within tumor cells, modulating the acidic, hypoxic microenvironment and generating reactive oxygen species (ROS), thereby enhancing anti-tumor efficacy. However, the efficient construction of hybrid nanosystems that combine the bioactivity of Ag with the microenvironmental regulation capabilities of MnO2 to achieve synergistic effects in tumor therapy remains a challenge. Based on this, the present invention proposes a method for preparing silver / manganese dioxide hybrid nanoparticles. By rationally designing the nanostructure and interface interaction, it gives them excellent tumor-targeted enrichment, microenvironment-responsive drug release and multi-mechanism synergistic anti-tumor properties, providing innovative ideas for the development of new anti-tumor nanomedicines. Summary of the Invention

[0003] To address the above-mentioned issues, the present invention proposes a method for preparing silver / manganese dioxide hybrid nanoparticles and their anti-tumor applications. The present invention provides the following technical solutions: A method for preparing silver / manganese dioxide hybrid nanoparticles comprises the following steps: (1) Synthesis of Ag@MnO2 hybrid core: Prepare 0.3M polyallylamine hydrochloride (PAH) and dissolve it in a volumetric flask. Prepare 0.04M AgNO3 and 0.02M KMnO4, dissolve them, and adjust the volume. Mix them and set aside. Add the mixture dropwise into the PAH solution (v:v = 8:1). React at 900 r / min in an ice bath for 15 min. Dilute the unreacted product in a 20KD dialysis bag for 4 h to obtain the Ag@MnO2 core. (2) Synthesis of Ag@MnO2-sis-L: Take 1 mL of dialyzed Ag@MnO2 solution, add 20 μL of 0.01 M citric acid solution, and adjust the Ag@MnO2 solution to acidic. Add an appropriate volume of siRNA and incubate for 10 min. Mix SM-102, CHO-HP, HSPC and DMG-PEG2000 (molar ratio = 18:16:4:1) and dissolve in an appropriate amount of anhydrous ethanol. Inject the lipid ethanol solution into the Ag@MnO2 solution, control the temperature at 55 ° C, rotate at 600 r / min, react for 30 min, and dialyze in a dialysis bag with a molecular weight cutoff of 20KD for 4 h to obtain Ag@MnO2-sis-L; (3) Synthesis of Ag@MnO2-sis-cL: DSPE-PEG2000-cRGD was added to the solution obtained in step (2), and the reaction was followed by dialysis to obtain Ag@MnO2-sis-cL.

[0004] Furthermore, the solvent of the PAH solution in step (1) is ultrapure water, and the fixed volume is 10 mL.

[0005] Furthermore, in step (1), the amount of the PAH solution added is 500 μL, and the amount of the AgNO 3 and KMnO 4 mixed solution added is 4 mL.

[0006] Furthermore, the concentration of the citric acid solution in step (2) is 0.01 M, and the pH is adjusted to 4.0-6.0.

[0007] Furthermore, in the lipid ethanol solution in step (2), the volume of ethanol is 100 μL.

[0008] Furthermore, the amount of DSPE-PEG2000-cRGD added in step (3) is 1 / 10 of the molar amount of DMG-PEG2000, and the reaction time is 30 min.

[0009] Furthermore, the silver / manganese dioxide hybrid nanoparticles (Ag@MnO2-sis-cL) obtained by the preparation method have a core-shell structure, wherein the core is Ag@MnO2 co-precipitated particles and the shell is a phospholipid layer encapsulating siRNA and SM-102, CHO-HP, HSPC, DMG-PEG2000 and DSPE-PEG2000-cRGD.

[0010] Furthermore, the application of the Ag@MnO2-sis-cL in the preparation of an integrated diagnostic and therapeutic preparation having both imaging and drug delivery functions.

[0011] Compared with the prior art, the present invention has the following beneficial effects: 1. The precise targeting capability of the drug of this invention is achieved through the phospholipid-modified cRGD targeting molecule, which can specifically recognize the highly expressed integrin αvβ3 on the surface of tumor cells, thereby achieving the enrichment of the contrast agent in tumor tissue, improving imaging resolution and lesion localization accuracy; 2. The drug of this invention exhibits excellent biocompatibility, specifically due to the fact that the phospholipid material (HSPC) and PEGylated lipids (DMG-PEG2000 and DSPE-PEG2000-cRGD) are both FDA-approved pharmaceutical excipients, which reduce immunogenicity and toxic side effects, ensuring in vivo circulation stability. The phospholipid coating improves particle dispersibility, maintaining a PDI of 0.23-0.25 (Table 1), ensuring in vivo stability. 3. The drug in this invention has the potential to integrate multifunctional diagnosis and treatment. The core is further loaded with siRNA, and the shell's targeted modification and phospholipid encapsulation impart drug delivery capabilities, enabling integrated diagnosis and treatment, providing a new platform for precision medicine. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 SEM images of Ag@MnO2, Ag@MnO2-sis-L, and Ag@MnO2-sis-cL prepared in the present invention; Figure 2 TEM images of Ag@MnO2, Ag@MnO2-sis-L, and Ag@MnO2-sis-cL prepared in the present invention; Table 1 shows the particle size, dispersion coefficient and potential of Ag@MnO2, Ag@MnO2-sis-L and Ag@MnO2-sis-cL prepared by the present invention. DETAILED DESCRIPTION Example 1

[0013] (1) 0.3 M PAH solution (500 μL) was placed in an ice bath, and a AgNO3 / KMnO4 mixture (4 mL) was added dropwise. The mixture was stirred for 15 min and dialyzed to obtain the Ag@MnO2 core (particle size 65.4 nm, PDI = 0.089, potential -45.57 mV). (2) Take 1 mL of dialyzed Ag@MnO2 solution, add 0.01 M citric acid to adjust the pH to acidic, add siRNA, and incubate for 10 min; (3) SM-102 (18 μmol), CHO-HP (16 μmol), HSPC (4 μmol), and DMG-PEG2000 (1 μmol) were dissolved in 100 μL of ethanol, injected into the Ag@MnO2 solution, reacted at 55°C for 30 min, and dialyzed to obtain Ag@MnO2-sis-L (particle size 80.7 nm, PDI = 0.247, potential 6.54 mV); (4) DSPE-PEG2000-cRGD (0.1 μmol) was added and dialyzed to obtain Ag@MnO2-sis-cL (particle size 84.4 nm, PDI = 0.234, potential 4.73 mV); Characterization analysis: Scanning electron microscopy (SEM, Figure 1) and transmission electron microscopy (TEM, Figure 2) showed that the Ag@MnO2 core was spherical with uniform particle size. After phospholipid encapsulation, a smooth shell was formed on the surface of the particles. There was no obvious change in the particle morphology after targeted modification, confirming the structural stability.

[0014] Table 1 preparation Particle size (nm) Potential (mV) PDI <![CDATA[Ag@MnO2]]> 65.44±1.27 -45.57±1.07 0.089±0.02 <![CDATA[Ag@MnO2-sis-L]]> 80.73±0.83 6.54±0.19 0.247±0.01 <![CDATA[Ag@MnO2-sis-c-L]]> 84.40±0.20 4.73±0.07 0.234±0.02

Claims

1. A method for preparing silver / manganese dioxide hybrid nanoparticles, characterized by: The following steps are involved: (1) Synthesis of Ag@MnO2 hybrid core: Prepare 0.3M polyallylamine hydrochloride (PAH) and dissolve it in a volumetric flask. Prepare 0.04M AgNO3 and 0.02M KMnO4, dissolve them, and adjust the volume. Mix them and set aside. Add the mixture dropwise into the PAH solution (v:v = 8:1). React at 900 r / min in an ice bath for 15 min. Dilute the unreacted product in a 20KD dialysis bag for 4 h to obtain the Ag@MnO2 core. (2) Synthesis of Ag@MnO2-sis-L: Take 1 mL of dialyzed Ag@MnO2 solution, add 20 μL of 0.01 M citric acid solution, and adjust the Ag@MnO2 solution to acidic. Add an appropriate volume of siRNA and incubate for 10 min. Mix SM-102, CHO-HP, HSPC and DMG-PEG2000 (molar ratio = 18:16:4:1) and dissolve in an appropriate amount of anhydrous ethanol. Inject the lipid ethanol solution into the Ag@MnO2 solution, control the temperature at 55 ° C, rotate at 600 r / min, react for 30 min, and dialyze in a dialysis bag with a molecular weight cutoff of 20KD for 4 h to obtain Ag@MnO2-sis-L; (3) Synthesis of Ag@MnO2-sis-cL: DSPE-PEG2000-cRGD was added to the solution obtained in step (2), and the reaction was followed by dialysis to obtain Ag@MnO2-sis-cL.

2. The method for preparing the Ag@MnO2 hybrid core according to claim 1, wherein: The solvent of the PAH solution in step (1) is ultrapure water, and the fixed volume is 10 mL.

3. The method for preparing the Ag@MnO2 hybrid core according to claim 1, wherein: In step (1), the amount of PAH solution added is 500 μL, and the amount of AgNO 3 and KMnO 4 mixed solution added is 4 mL.

4. The method for preparing the Ag@MnO2 hybrid core according to claim 1, wherein: The concentration of the citric acid solution in step (2) is 0.01 M, and the pH is adjusted to 4.0-6.

0.

5. The preparation method according to claim 1, wherein: In the lipid ethanol solution in step (2), the volume of ethanol is 100 μL.

6. The preparation method according to claim 1, wherein: The amount of DSPE-PEG2000-cRGD added in step (3) is 1 / 10 of the molar amount of DMG-PEG2000, and the reaction time is 30 min.

7. Silver / manganese dioxide hybrid nanoparticles (Ag@MnO2-sis-cL) obtained by the preparation method according to any one of claims 1 to 6, characterized in that: The contrast agent has a core-shell structure, wherein the core is Ag@MnO2 co-precipitated particles and the shell is a phospholipid layer encapsulating siRNA and SM-102, CHO-HP, HSPC, DMG-PEG2000 and DSPE-PEG2000-cRGD.

8. Use of the Ag@MnO2-sis-cL according to claim 1 in the preparation of an integrated diagnostic and therapeutic preparation with both imaging and drug delivery functions.

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