Preparation method and application of thorny structure nanoparticles

Abstract

The invention discloses a preparation method and application of thorny structure nanoparticles, and relates to the technical field of biological medicines. The nano particle comprises a multi-thorn carbon nano particle, a manganese dioxide shell layer deposited on the surface of the multi-thorn carbon nano particle, and a fluorescence imaging molecule and a targeting ligand molecule of which the surfaces are modified with the manganese dioxide shell layer, the obtained multi-thorn-structure nano particles have uniform particle size distribution, the photothermal conversion efficiency and the capability of being endocytosed by cells are remarkably improved through the thorn-shaped structures with the dense surfaces, and the multi-thorn-structure nano particles have the synergistic enhancement treatment effect on various types of tumors.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a preparation method and application of nanoparticles with a spiny structure. Background Art

[0002] Nanoscale diagnostic and therapeutic agents have greatly promoted the development of modern medicine. These nanosystems can maximize therapeutic effects and reduce side effects by delivering therapeutic drugs, proteins or imaging agents to the lesion area. Therefore, many studies are devoted to designing diagnostic and therapeutic nanosystems with high enrichment ability and specific targeting to enhance their interaction with biological systems, specific organs or target cells. A large number of studies have shown that the surface chemical properties of nanoparticles, including hydrophilicity, charge, targeting ligands, protein corona and functional group density, have an important influence on their cellular uptake, intracellular accumulation, biological response and toxicity. For example, some work focuses on the effect of the size and geometry of nanomaterials on the above behaviors. Studies have shown that spherical nanoparticles with a diameter of about 100 nm are easily engulfed by cells, and rod-shaped nanoparticles with a high aspect ratio also show great advantages in endocytosis. However, there are relatively few studies on the endocytosis of nanomaterials with more complex geometries. Due to the key role of cellular uptake efficiency in the field of nanomedicine, exploring how to optimize the internalization mechanism of nanomaterials with complex geometries is of great significance to promoting the advancement of nanotechnology.

[0003] Some viruses, such as coronaviruses, have many spike-like structures on their surfaces. By combining these structures with cell membranes, receptor-specific interactions can be multiplied, resulting in high viral infectivity. Tumor photothermal therapy is an emerging, non-invasive cancer treatment method that uses photothermal conversion materials to absorb light energy and convert it into heat energy, thereby generating a high temperature effect locally in the tumor, leading to tumor cell necrosis or apoptosis. This method has a high degree of spatial selectivity and controllability, and can act precisely on tumor tissues while causing relatively little damage to surrounding normal tissues. Ideal photothermal conversion materials should have good biocompatibility, high photothermal conversion efficiency, and stable physical and chemical properties to ensure that they can function safely and effectively in vivo. With the rapid development of nanotechnology, common photothermal conversion materials, such as gold nanoparticles, carbon nanotubes, graphene and its derivatives, have shown great application potential in the field of tumor photothermal therapy due to their unique physical and chemical properties. However, the existing photothermal conversion materials still have certain limitations, such as insufficient cell uptake rate, insufficient photothermal effect, or biocompatibility needs to be improved. Therefore, the development of new, efficient, safe and reliable photothermal conversion materials remains a key issue that needs to be urgently addressed in the current field of tumor photothermal therapy.

[0004] Based on this, the present invention provides a thorny hollow nanoparticle, which can achieve a higher cellular uptake rate and better photothermal anti-tumor effect compared with similar spherical particles. Summary of the invention

[0005] In view of the shortcomings of the above-mentioned background technology, the present invention mainly aims at the limitations of the existing photothermal conversion materials, such as insufficient cell uptake rate, insufficient photothermal effect or biocompatibility to be improved. The present invention provides a preparation method and application of spiny structured nanoparticles. The nanoparticles are spiny hollow nanoparticles with high cell uptake efficiency and stronger photothermal conversion efficiency. By coupling the surface with near-infrared (NIR) dyes, the cell uptake of nanoparticles can be quantified by measuring the intracellular manganese concentration and fluorescence intensity. Depending on the cell type, the cell uptake rate of spiny particles is 2.3 to 8.8 times higher than that of spherical particles. In addition, due to the cone structure of the nanospikes, the spiny particles show a better photothermal effect than spherical particles. After further modification of the targeting ligand, the spiny particles can effectively accumulate in the tumor site, and degrade and release signal molecules with magnetic resonance activity under the mild acidic and reducing microenvironment inside the tumor cells. Under the guidance of magnetic resonance imaging (MRI) and near-infrared fluorescence imaging, the nanoparticles can achieve enhanced photothermal therapy to ablate tumors.

[0006] The first object of the present invention is to provide a spiny structure nanoparticle, which comprises a spiny carbon nanoparticle, a manganese dioxide shell layer deposited on the surface of the spiny carbon nanoparticle, and a fluorescent imaging molecule and a targeting ligand molecule that surface-modify the manganese dioxide shell layer; The spiny carbon nanoparticles are hollow structures; The fluorescent imaging molecule contains a carboxyl group; The targeting ligand molecule is a tumor cell-specific targeting molecule.

[0007] Preferably, the spiny carbon nanoparticles are prepared according to the following steps: Using silicon dioxide nanoparticles as core templates, dopamine hydrochloride and tetraethyl silicate were copolymerized on the particle surface to form a shell layer, followed by a carbonization process to form a carbon-silicon composite shell layer, and then etching to remove the silicon element to obtain hollow-structured spiny carbon nanoparticles; The mass ratio of dopamine hydrochloride to tetraethyl silicate is 1:5-10.

[0008] Preferably, the particle size of the spiny carbon nanoparticles is 150-200 nm; and the particle size of the silicon dioxide is 50-120 nm.

[0009] Preferably, the thickness of the manganese dioxide shell is 20-30 nm, and the loading amount is 5-15%; The modification amount of the fluorescent imaging molecule accounts for 1-2% of the mass of the spiny carbon nanoparticles; The modification amount of the targeting ligand molecule accounts for 2-4% of the mass of the spiny carbon nanoparticles.

[0010] Preferably, the fluorescent imaging molecules include molecules with emission wavelengths in the near-infrared region such as Cypate, Cyanine7 or IR820; The targeting ligand molecule is an antibody, polysaccharide, P-selectin or a protein that can specifically target tumor cells including transferrin, folic acid or RGD polypeptide.

[0011] The second object of the present invention is to provide a method for preparing spiny structure nanoparticles, comprising the following steps: Preparation of spiny carbon nanoparticles with hollow structures; The spiny carbon nanoparticles are uniformly dispersed in a water solvent, potassium permanganate is added, and the mixture is reacted at 75-85°C for 8-12 hours to obtain particles with a manganese dioxide shell layer deposited on the surface; The particles with a manganese dioxide shell layer deposited on the surface are uniformly dispersed in ethanol, 3-aminopropyltriethoxysilane is added, and the mixture is reacted at 55-65° C. for 12-24 hours to obtain particles with amino groups modified on the surface; The particles with amino groups modified on the surface are uniformly dispersed in DMF, fluorescent imaging molecules and polyethylene glycol are added, and then targeting ligand molecules are added, and mixed to react to obtain spiny structure nanoparticles.

[0012] Preferably, the mass ratio of the potassium permanganate to the spiny carbon nanoparticles is 1:3-7.

[0013] Preferably, the polyethylene glycol molecules include two types, one of which has an NHS ester at one end of the molecular end group and a maleimide at the other end, and a molecular weight of 2000; the other has an NHS ester at one end of the molecular end group and a methoxy group at the other end, and a molecular weight of 2000; the mass ratio of the two polyethylene glycols is 1:4.

[0014] Preferably, the amount of 3-aminopropyltriethoxysilane added is 5-15% of the mass of manganese dioxide deposited on the surface. The third object of the present invention is to provide a use of spiny structure nanoparticles in the preparation of tumor drugs.

[0015] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a preparation method and application of spiny structure nanoparticles. The obtained multi-thorn structure nanoparticles have a uniform particle size distribution, and the dense thorn-like structure on the surface significantly improves the photothermal conversion efficiency and the ability to be internalized by cells, and has a synergistically enhanced therapeutic effect on various types of tumors.

[0016] The present invention modifies polyethylene glycol molecules with long blood circulation function and protein molecules with tumor cell targeting by a simple method. The modification of polyethylene glycol molecules significantly improves the circulation stability of nanoparticles in the blood, effectively reduces the possibility of being recognized and cleared by the immune system, thereby extending its half-life in the body. This not only allows the drug to remain in the effective concentration range for a longer time, but also improves the bioavailability of the drug. The introduction of protein molecules with tumor cell targeting function, such as antibody fragments, receptor ligands, etc., enables nanoparticles to specifically recognize and bind to the surface of tumor cells. This high degree of targeting reduces the exposure of the drug to non-target tissues, reduces side effects, and enhances the therapeutic effect.

[0017] The present invention connects manganese dioxide and near-infrared fluorescent molecules to the surface of particles through simple chemical modification, giving the system dual-mode imaging functions of magnetic resonance imaging and fluorescence imaging. Under the guidance of imaging, the distribution of particles in the body and the enrichment of lesion areas can be accurately judged, thereby providing more comprehensive and accurate tumor information, helping to achieve precise positioning of tumors and improve the accuracy and safety of surgical resection. The integrated diagnosis and treatment design of the spiny structured nanoparticles can diagnose the disease in real time and accurately and treat it simultaneously, so that the efficacy can be monitored during the treatment process and the dosage regimen can be adjusted at any time, which is conducive to achieving the best treatment effect. Especially for the treatment of diseases such as tumors, this real-time and accurate monitoring and regulation capability can significantly improve the treatment effect and reduce the pain of patients.

[0018] The present invention adopts a simple and efficient preparation and modification method to prepare hollow nanoparticles with a spiny structure, realize magnetic resonance / fluorescence dual-mode imaging as well as enhanced cellular endocytosis effect and synergistic photothermal therapeutic effect. The particle preparation process can be scaled up for production, which is of great significance for promoting the clinical transformation and wide application of nanomedicines.

[0019] The photothermal conversion ability of the nanoparticles provided by the present invention in the photothermal treatment of tumors largely determines its therapeutic effect. In addition, the efficacy of the nanoagent is also determined by the effect of being internalized by tumor cells. The present invention has found that carbon nanoparticles with a thorny structure on the surface have a stronger photothermal conversion effect. Under 808 nm laser irradiation, the dispersion temperature can be raised to about 60 degrees Celsius within 5 minutes. At the same time, the thorny structure significantly increases the content of particles taken up by tumor cells, and the enrichment in tumor cells is increased by about 5-8 times compared with the same type of particles without thorny structure. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 These are transmission electron microscopy images of spiny / spineless hollow nanoparticles; among them, (a) is a transmission electron microscopy image of spiny hollow nanoparticles; (b) is a transmission electron microscopy image of thornless hollow nanoparticles.

[0021] Figure 2 X-ray diffraction patterns of spiny / spineless hollow nanoparticles.

[0022] Figure 3 Thermogravimetric analysis of spiny / spineless hollow nanoparticles.

[0023] Figure 4 The magnetic resonance T1 signal of the spiny / spineless hollow nanoparticles changes over time under pH 5 or 10 mM GSH.

[0024] Figure 5 The graph shows the near-infrared fluorescence (810 nm) signal of the spiny / spineless hollow nanoparticles changing over time at pH 5 or 10 mM GSH.

[0025] Figure 6 The figure shows the content of spiny / spineless hollow nanoparticles in tumor cells A549 at different concentrations.

[0026] Figure 7 The spiny / spineless hollow nanoparticle dispersion was exposed to near-infrared light (808 nm, 1 W / cm -2） The dispersion temperature changes with irradiation time under irradiation.

[0027] Figure 8 This is a graph showing the temperature changes in the tumor area of ​​mice as a function of irradiation time after injection of the spiny / spineless hollow nanoparticle dispersion sample.

[0028] Fig. 9 These are pictures of mouse tumors and tumor mass removed 20 days after photothermal therapy with spiny / spineless hollow nanoparticles.

[0029] Fig.10 This is a graph showing weight changes during photothermal therapy of mouse tumors. DETAILED DESCRIPTION

[0030] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below in conjunction with specific embodiments and drawings, but the embodiments are not intended to limit the present invention.

[0031] The present invention realizes the controllable preparation of multi-spiny nanoparticles through an innovative chemical reaction pathway, combined with the regulation of specific reactant ratios and modified molecule ratios. This method is not only simple to operate and has mild conditions, but also can accurately control the size of the particles and the density and length of the surface thorn-like structures. The obtained multi-spiny nanoparticles have a uniform particle size distribution, and the dense thorn-like structures on the surface significantly improve the photothermal conversion efficiency and the ability to be internalized by cells, and have a synergistically enhanced therapeutic effect on various types of tumors.

[0032] In order to achieve the above-mentioned object, the first aspect of the present invention provides a spiny structure nanoparticle, which comprises a spiny carbon nanoparticle, a manganese dioxide shell layer deposited on the surface of the spiny carbon nanoparticle, and a fluorescent imaging molecule and a targeting ligand molecule that surface-modify the manganese dioxide shell layer; The spiny carbon nanoparticles are hollow structures; the fluorescent imaging molecules contain carboxyl groups; and the targeting ligand molecules are tumor cell-specific targeting molecules.

[0033] The spiny structured nanoparticles provided by the present invention are mainly used to enhance cellular uptake and photothermal therapy effects, including spiny carbon nanoparticles for converting light energy into thermal energy, a manganese dioxide shell layer deposited on the surface of the carbon nanoparticles, a fluorescent imaging molecule modified with the manganese dioxide shell layer, and a targeting ligand molecule.

[0034] The spiny carbon nanoparticles are prepared according to the following steps: Using silicon dioxide nanoparticles as core templates, dopamine hydrochloride and tetraethyl silicate were copolymerized on the particle surface to form a shell layer, followed by a carbonization process to form a carbon-silicon composite shell layer, and then etching to remove the silicon element to obtain hollow-structured spiny carbon nanoparticles; The mass ratio of dopamine hydrochloride to tetraethyl silicate is 1:5-10.

[0035] During etching, a sodium hydroxide solution is used to remove the silicon element.

[0036] The particle size of the spiny carbon nanoparticles is 150-200 nm, and the particle size of the silicon dioxide is 50-120 nm. The spiny structure morphology can be controlled by adjusting the ratio of dopamine hydrochloride and tetraethyl silicate.

[0037] The thickness of the manganese dioxide shell is 20-30 nm, and the loading is 5-15%; The modification amount of the fluorescent imaging molecule accounts for 1-2% of the mass of the spiny carbon nanoparticles; The modification amount of the targeting ligand molecule accounts for 2-4% of the mass of the spiny carbon nanoparticles.

[0038] Preferably, the thickness of the manganese dioxide shell is 25 nm, and the loading amount is 10%; the modification amount of the fluorescent imaging molecule accounts for 1.5% of the mass of the nanoparticle; and the modification amount of the targeting ligand molecule accounts for 3% of the mass of the nanoparticle.

[0039] The fluorescent imaging molecules include: Cypate, Cyanine7, IR820 and other molecules with emission wavelengths in the near-infrared region.

[0040] The targeting ligand molecules include: transferrin, folic acid, RGD polypeptide and other proteins, antibodies, polysaccharides, and P-selectin that can specifically target tumor cells.

[0041] The particle size of the silicon dioxide nanoparticles is about 100 nm, and silicon dioxide nanoparticles of different particle sizes can be used as the core to adjust the particle size of the prepared spiny carbon nanoparticles.

[0042] The manganese dioxide shell deposition specifically includes the following steps: adding potassium permanganate to a fully dispersed suspension of spiny nanoparticles, stirring vigorously, and reacting at 80 degrees Celsius for 10 hours to obtain composite manganese dioxide spiny hollow nanoparticles. The mass ratio of potassium permanganate to spiny carbon nanoparticles can be adjusted to adjust the content of manganese dioxide on the surface of the spiny nanoparticles, wherein the optimal ratio is 1:5.

[0043] The fluorescent molecule is a near-infrared fluorescent imaging molecule with an excitation wavelength between 600-1000 nm and an emission wavelength above 800 nm, and contains a carboxyl group.

[0044] The targeting ligand molecule is a tumor cell-specific targeting molecule, including antibodies, membrane proteins, polypeptides, polysaccharides, and the like.

[0045] The manganese dioxide shell is modified with fluorescent molecules and targeting molecules by the following method: firstly, the prickly hollow nanoparticles of composite manganese dioxide are dispersed in an ethanol solution, 3-aminopropyltriethoxysilane is added, and the reaction is carried out at 60 degrees Celsius for 24 hours to modify the surface of the particles with amino groups, and the fluorescent molecules are connected by the reaction between the amino groups and the activated carboxyl groups, and then the polyethylene glycol molecules are connected to the surface of the particles, and finally the targeting molecules are added to obtain the prickly hollow nanoparticles carrying the fluorescent molecules and the targeting molecules.

[0046] A second aspect of the present invention provides a method for preparing spiny structure nanoparticles, comprising the following steps: Preparation of spiny carbon nanoparticles with hollow structures; The spiny carbon nanoparticles are uniformly dispersed in a water solvent, potassium permanganate is added, and the mixture is reacted at 75-85°C for 8-12 hours to obtain particles with a manganese dioxide shell layer deposited on the surface; The particles with a manganese dioxide shell layer deposited on the surface are uniformly dispersed in ethanol, 3-aminopropyltriethoxysilane is added, and the mixture is reacted at 55-65° C. for 12-24 hours to obtain particles with amino groups modified on the surface; The particles with amino groups modified on the surface are uniformly dispersed in DMF, fluorescent imaging molecules and polyethylene glycol are added, and then targeting ligand molecules are added, and mixed to react to obtain spiny structure nanoparticles.

[0047] Wherein, the mass ratio of the potassium permanganate to the spiny carbon nanoparticles is 1:3-7.

[0048] The polyethylene glycol molecules include two types, one of which has an NHS ester at one end of the molecular end group and a maleimide at the other end, and has a molecular weight of 2000; the other has an NHS ester at one end of the molecular end group and a methoxy group at the other end, and has a molecular weight of 2000; the mass ratio of the two polyethylene glycols is 1:4.

[0049] The amount of 3-aminopropyltriethoxysilane added is 10% of the mass of manganese dioxide particles deposited on the surface.

[0050] A third aspect of the present invention provides a use of spiny structure nanoparticles in the preparation of tumor drugs.

[0051] It should be noted that the experimental methods used in the present invention are all conventional methods unless otherwise specified; the reagents and materials used are all commercially available unless otherwise specified.

[0052] Example 1 This example is a preparation method and application verification of hollow nanoparticles with a spiny structure, which mainly includes the following steps: (1) Preparation of silica core template: 1.34 mL of tetraethyl silicate and 1.2 mL of ammonia water were dissolved in 25% ethanol, stirred at room temperature for 24 hours, and then centrifuged and washed with water to obtain silica nanoparticles.

[0053] (2) Preparation of hollow carbon nanoparticles with a spiny structure. 100 mg of the above-mentioned silica nanoparticles, 1.5 mL of ammonia water and 5 mL of deionized water were added to 35 mL of ethanol, followed by 100 mg of dopamine hydrochloride and 1 mL of tetraethyl silicate. The reaction was carried out at room temperature for 24 hours to obtain nanoparticles with a shell layer of dopamine-composite silica. After centrifugal washing, the particles were placed in a nitrogen atmosphere and the ambient temperature was raised to 550 degrees Celsius at a rate of 1 degree Celsius per minute for 5 hours. This step can decompose and carbonize the polydopamine on the surface. Then, the carbonized nanoparticles were placed in a sodium hydroxide solution with a sodium hydroxide concentration of 2M. After stirring at room temperature for a period of time, hollow carbon nanoparticles with a spiny structure were obtained.

[0054] (3) Preparation of hollow carbon nanoparticles without spiny structures. The steps are basically the same as (2), except that 100 mg of dopamine hydrochloride and 1 mL of tetraethyl silicate are added instead of only 200 mg of dopamine hydrochloride.

[0055] (4) Manganese dioxide deposition and surface modification of spiny structured nanoparticles. 100 mg of the above-mentioned spiny structured hollow carbon nanoparticles or non-spiny structured hollow carbon nanoparticles were dispersed in 50 mL of deionized water, and 20 mg of potassium permanganate was added under vigorous stirring. The mixture was reacted at 80 degrees Celsius for 10 hours to obtain nanoparticles with manganese dioxide deposited on the surface, such as Figure 1 As shown in the transmission electron microscope image, the particle size of the prepared particles is about 180 nm. Take 100 mg of the above particles and disperse them in 100 mL of ethanol, add 50 μL of 3-aminopropyltriethoxysilane, and react at 60 degrees Celsius for 24 hours to obtain particles with surface modified with amino groups. Dissolve 20 mg of the above particles in 20 mL of DMF, add 5 mg of NHS ester-activated Cypate (a near-infrared fluorescent molecule), 4 mgMal-PEG-NHS and 12 mg mPEG-NHS, and stir at room temperature for 24 hours to obtain PEG and fluorescent modified spiky structure nanoparticles, that is, spiky structure nanoparticles (spiky-C@MnO2) and spherical structure nanoparticles (spherical-C@MnO2) are obtained respectively.

[0056] (5) Performance verification of spiny nanoparticles. The prepared spiny / non-spiny nanoparticles were characterized by XRD. Figure 2 It can be seen that there is a manganese dioxide shell on the surface of the particles; the particles were tested using a thermogravimetric analyzer. Figure 3 It can be seen that the manganese content in the two particles is about 10%; the T1 signal intensity of the two particles was detected using magnetic resonance equipment. Figure 4 It can be seen that under acidic conditions, the deposited manganese dioxide of the two particles will degrade to produce divalent manganese ions, which significantly enhances the T1 signal. In addition, the magnetic resonance signals of the nanoparticles with and without thorns are similar, indicating that the endocytosis of the two particles can be determined by detecting the strength of the T1 signal in the cells. The near-infrared fluorescence intensity signals of the two particles were detected using a fluorescence spectrometer. Figure 5 It can be seen that under acidic conditions, the fluorescence intensity increases due to particle degradation. At the same time, the fluorescence signals of nanoparticles with / without thorns are similar, indicating that the endocytosis of the two particles can be determined by detecting the strength of the fluorescence signal in the cells.

[0057] (5) Application of spiny nanoparticles in improving cell endocytosis. The nanoparticles with or without spiny structures prepared in (1)-(4) were added to cells at a density of 2×10 5In each well of the culture plate, the cells are human lung cancer A549 cells, and the particle concentrations are 5, 10, 20, and 50 μg / mL, respectively. The intracellular manganese ion concentration is measured to quantitatively analyze the particle endocytosis. Figure 6 As shown in , at the same particle concentration, the amount of nanoparticles with a spiny structure internalized by A549 is about 8 times that of nanoparticles without a spiny structure, indicating that the particles have a good function of enhancing cell endocytosis.

[0058] Example 2 This example is a method for optimizing spiny nanoparticles and evaluating the synergistic enhancement of photothermal therapy effects. 20 mg of the PEG and fluorescently modified spiny nanoparticles obtained in Example 1 and 2 mg of thiolated transferrin were mixed at room temperature for 24 hours to obtain spiny nanoparticles with tumor cell targeting function.

[0059] (1) Photothermal performance verification of spiny nanoparticles. The dispersion of spiny and non-spiny nanoparticles with a concentration of 50 μg / mL was placed at 808 nm and 1 W / cm -2 The temperature of the dispersion was monitored and recorded in real time using a thermal imager. Figure 7 As shown, the nanoparticles with a thorny structure exhibit better photothermal effects than the nanoparticles without a thorny structure, and can raise the temperature of the dispersion to about 60 degrees Celsius within 5 minutes.

[0060] Among them, PBS (physiological saline) is Figure 7-10 The control group of mice was injected with PBS via the tail vein. Spiky-C@MnO2-Tf (spiky nanoparticles with fluorescent molecules, PEG and transferrin on the surface) was attached Figure 7-10 The spherical-C@MnO2-Tf (spherical-free nanoparticles with fluorescent molecules, PEG and transferrin on the surface) was injected into the tail vein of the mice. Figure 7-10 The middle represents the experimental group in which the mice were injected with spikeless nanoparticles through the tail vein.

[0061] (2) Verification of the photothermal therapeutic effect of spiny nanoparticles in small animal tumor models. A549 mouse transplant tumor model was constructed. When the tumor volume grew to about 100 mm 3 PBS solution, thornless nanoparticles and thorny nanoparticle dispersions were injected through the tail vein at a dose of 5 mg / kg. Four hours later, the tumor area was irradiated with 808 nm near-infrared light for 5 minutes at a laser power of 1 W / cm -2 , using a thermal imager to record temperature changes in the tumor area in real time, such as Figure 8As shown in the figure, the temperature of the tumor area of ​​mice injected with thorny nanoparticles reached about 50 degrees Celsius after irradiation, while that of the thornless group was only 30 degrees Celsius, indicating that the thorny structure on the surface of the particles not only increased the enrichment in the tumor area, but also had a better photothermal conversion effect, thereby obtaining a higher temperature in the lesion area. Subsequently, a vernier caliper was used to measure the tumor volume of the mice every other day and record it. On the 20th day, the mice were euthanized and the tumor tissue was removed, photographed, weighed and recorded. Fig. 9 It can be seen that the tumors of mice injected with the spiny structure nanoparticle samples can be completely eliminated, proving that the particles have excellent photothermal therapeutic effects.

[0062] (3) Biosafety evaluation of spiny nanoparticles. In the mouse tumor model, in addition to measuring the tumor volume every other day, the weight of the mice was also weighed and recorded. Fig.10 It can be seen that the weight changes of mice in the experimental group are not significantly different from those in the control group, indicating that the spiny structured nanoparticles have good biosafety.

[0063] The present invention describes preferred embodiments and their effects. However, those skilled in the art may make additional changes and modifications to these embodiments once they are aware of the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0064] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A spiny nanoparticle, characterized in that: The nanoparticles include prickly carbon nanoparticles, a manganese dioxide shell layer deposited on the surface of the prickly carbon nanoparticles, and fluorescent imaging molecules and targeting ligand molecules that modify the surface of the manganese dioxide shell layer; The spiny carbon nanoparticles are hollow structures; The fluorescent imaging molecule contains a carboxyl group; The targeting ligand molecule is a tumor cell-specific targeting molecule.

2. The spiny structure nanoparticle according to claim 1, characterized in that: The spiny carbon nanoparticles are prepared according to the following steps: Using silicon dioxide nanoparticles as core templates, dopamine hydrochloride and tetraethyl silicate were copolymerized on the particle surface to form a shell layer, followed by a carbonization process to form a carbon-silicon composite shell layer, and then etching to remove the silicon element to obtain hollow-structured spiny carbon nanoparticles; The mass ratio of dopamine hydrochloride to tetraethyl silicate is 1:5-10.

3. The spiny structure nanoparticle according to claim 2, characterized in that: The particle size of the spiny carbon nanoparticles is 150-200 nm; the particle size of the silicon dioxide is 50-120 nm.

4. The spiny structure nanoparticle according to claim 1, characterized in that: The thickness of the manganese dioxide shell is 20-30 nm, and the loading is 5-15%; The modification amount of the fluorescent imaging molecule accounts for 1-2% of the mass of the spiny carbon nanoparticles; The modification amount of the targeting ligand molecule accounts for 2-4% of the mass of the spiny carbon nanoparticles.

5. The spiny structure nanoparticle according to claim 1, characterized in that: The fluorescent imaging molecules include molecules with emission wavelengths in the near-infrared region, such as Cypate, Cyanine7 or IR820; The targeting ligand molecule is an antibody, polysaccharide, P-selectin or a protein that can specifically target tumor cells including transferrin, folic acid or RGD polypeptide.

6. A method for preparing the spiny structure nanoparticles according to any one of claims 1 to 5, characterized in that: The following steps are involved: Preparation of spiny carbon nanoparticles with hollow structures; The spiny carbon nanoparticles are uniformly dispersed in a water solvent, potassium permanganate is added, and the mixture is reacted at 75-85°C for 8-12 hours to obtain particles with a manganese dioxide shell layer deposited on the surface; The particles with a manganese dioxide shell layer deposited on the surface are uniformly dispersed in ethanol, 3-aminopropyltriethoxysilane is added, and the mixture is reacted at 55-65° C. for 12-24 hours to obtain particles with amino groups modified on the surface; The particles with amino groups modified on the surface are uniformly dispersed in DMF, fluorescent imaging molecules and polyethylene glycol are added, and then targeting ligand molecules are added, and mixed to react to obtain spiny structure nanoparticles.

7. The method for preparing the spiny structure nanoparticles according to claim 6, characterized in that: The mass ratio of the potassium permanganate to the spiny carbon nanoparticles is 1:3-7.

8. The method for preparing spiny structure nanoparticles according to claim 6, characterized in that: The polyethylene glycol molecules include two types, one of which has an NHS ester at one end of the molecular end group and a maleimide at the other end, and has a molecular weight of 2000; the other has an NHS ester at one end of the molecular end group and a methoxy group at the other end, and has a molecular weight of 2000; the mass ratio of the two polyethylene glycols is 1:

4.

9. The method for preparing spiny structure nanoparticles according to claim 6, characterized in that: The amount of 3-aminopropyltriethoxysilane added is 5-15% of the mass of manganese dioxide deposited on the surface.

10. Use of the spiny structure nanoparticles according to any one of claims 1 to 5 in the preparation of tumor drugs.

Images