Preparation method and application of spiny structured nanoparticles

Abstract

The present invention discloses a method for preparing and applying spiny nanoparticles, which relate to the field of biomedicine. The nanoparticles comprise spiny carbon nanoparticles, a manganese dioxide shell deposited on the surface of the spiny carbon nanoparticles, and fluorescent imaging molecules and targeting ligand molecules that modify the manganese dioxide shell. The spiny nanoparticles have a uniform particle size distribution, and the densely packed spiny surface structures significantly enhance photothermal conversion efficiency and cellular endocytosis, resulting in synergistically enhanced therapeutic effects on various tumor types.

Description

Technical Field

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

[0002] Nanoscale diagnostic and therapeutic agents have significantly advanced modern medicine. These nanosystems can deliver therapeutic drugs, proteins, or imaging agents to diseased areas, maximizing therapeutic efficacy and minimizing side effects. Consequently, extensive research is focused on designing diagnostic and therapeutic nanosystems with high enrichment and specific targeting capabilities to enhance their interactions with biological systems, specific organs, or target cells. Numerous studies have demonstrated that the surface chemical properties of nanoparticles, including hydrophilicity, charge, targeting ligands, protein corona, and density of functional groups, significantly influence their cellular uptake, intracellular accumulation, biological responses, and toxicity. For example, some research has focused on the influence of nanomaterial size and geometry on these behaviors. Studies have shown that spherical nanoparticles with diameters of approximately 100 nm are readily endocytosed, while rod-shaped nanoparticles with high aspect ratios also exhibit significant advantages in endocytosis. However, relatively little research has been conducted on the endocytosis of nanomaterials with more complex geometries. Given the critical role of cellular uptake efficiency in nanomedicine, exploring how to optimize the internalization mechanisms of nanomaterials with complex geometries is crucial for advancing nanotechnology.

[0003] Some viruses, such as coronaviruses, have numerous spike-like structures on their surfaces. These structures bind to cell membranes, multiplying receptor-specific interactions and leading to high viral infectivity. Tumor photothermal therapy is an emerging, non-invasive cancer treatment. It utilizes photothermal conversion materials to absorb light energy and convert it into heat, generating a localized hyperthermia in the tumor, leading to tumor cell necrosis or apoptosis. This approach offers high spatial selectivity and controllability, enabling precise targeting of tumor tissue with minimal damage to surrounding normal tissue. Ideal photothermal conversion materials should possess excellent biocompatibility, high photothermal conversion efficiency, and stable physicochemical properties to ensure safe and effective in vivo function. With the rapid development of nanotechnology, common photothermal conversion materials, such as gold nanoparticles, carbon nanotubes, graphene, and their derivatives, have shown great potential for application in tumor photothermal therapy due to their unique physical and chemical properties. However, currently available photothermal conversion materials still have limitations, such as insufficient cellular uptake, insignificant photothermal effects, and the need for improved biocompatibility. Therefore, the development of new, efficient, safe and reliable photothermal conversion materials remains a key issue that needs to be urgently addressed in the field of tumor photothermal therapy.

[0004] Based on this, the present invention provides a spiny 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] To address the shortcomings of the aforementioned background technologies, the present invention primarily addresses the limitations of existing photothermal conversion materials, such as insufficient cellular uptake, insignificant photothermal effects, and the need for improved biocompatibility. The present invention provides a method for preparing and applying spiny nanoparticles. These hollow nanoparticles exhibit high cellular uptake efficiency and enhanced photothermal conversion efficiency. By surface-coupling with a near-infrared (NIR) dye, cellular uptake of the nanoparticles can be quantified by measuring intracellular manganese concentration and fluorescence intensity. Depending on the cell type, the cellular uptake of the spiny nanoparticles is 2.3 to 8.8 times higher than that of their spherical counterparts. Furthermore, due to the conical structure of the nanospikes, the spiny nanoparticles exhibit superior photothermal effects compared to spherical nanoparticles. After further modification with targeting ligands, the spiny nanoparticles can effectively accumulate in tumors and, in the mildly acidic and reducing microenvironment within tumor cells, degrade and release magnetic resonance-active signaling molecules. Guided by magnetic resonance imaging (MRI) and near-infrared fluorescence imaging, the nanoparticles can achieve enhanced photothermal therapy for tumor ablation.

[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;

[0007] The spiny carbon nanoparticles are hollow structures;

[0008] The fluorescent imaging molecule contains a carboxyl group;

[0009] The targeting ligand molecule is a tumor cell-specific targeting molecule.

[0010] Preferably, the spiny carbon nanoparticles are prepared according to the following steps:

[0011] Using silica nanoparticles as core templates, dopamine hydrochloride and tetraethyl silicate are copolymerized on the particle surface to form a shell layer. This is followed by a carbonization process to form a carbon-silicon composite shell layer. The silicon element is then etched away to obtain hollow-structured spiny carbon nanoparticles.

[0012] The mass ratio of dopamine hydrochloride to tetraethyl silicate is 1:5-10.

[0013] Preferably, the particle size of the spiny carbon nanoparticles is 150-200 nm; the particle size of the silicon dioxide is 50-120 nm.

[0014] Preferably, the thickness of the manganese dioxide shell is 20-30 nm, and the loading amount is 5-15%;

[0015] The modified amount of the fluorescent imaging molecule accounts for 1-2% of the mass of the spiny carbon nanoparticles;

[0016] The modified amount of the targeting ligand molecule accounts for 2-4% of the mass of the spiny carbon nanoparticles.

[0017] Preferably, the fluorescent imaging molecules include molecules with emission wavelengths in the near-infrared region such as Cypate, Cyanine7 or IR820;

[0018] 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.

[0019] A second object of the present invention is to provide a method for preparing spiny structured nanoparticles, comprising the following steps:

[0020] Preparation of spiny carbon nanoparticles with hollow structures;

[0021] The spiny carbon nanoparticles are uniformly dispersed in an aqueous 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 deposited on the surface.

[0022] The particles with a manganese dioxide shell deposited on the surface were uniformly dispersed in ethanol, 3-aminopropyltriethoxysilane was added, and the mixture was reacted at 55-65°C for 12-24 hours to obtain particles with amino groups modified on the surface;

[0023] The obtained particles modified with amino groups 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.

[0024] Preferably, the mass ratio of the potassium permanganate to the spiny carbon nanoparticles is 1:3-7.

[0025] Preferably, the polyethylene glycol molecules include two types, one of which has an NHS ester at one end and a maleimide at the other end, and has a molecular weight of 2000; the other has an NHS ester at one end 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.

[0026] Preferably, the amount of 3-aminopropyltriethoxysilane added is 5 to 15% of the mass of manganese dioxide deposited on the surface.

[0027] The third object of the present invention is to provide an application of spiny structure nanoparticles in the preparation of tumor drugs.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention provides a preparation method and application of spiny structure nanoparticles.

[0030] The spiny nanoparticles obtained by the present invention have a uniform particle size distribution, and the dense spiny structures on the surface significantly improve the photothermal conversion efficiency and the ability to be endocytosed by cells, and have a synergistically enhanced therapeutic effect on various types of tumors.

[0031] The present invention uses a simple method to modify polyethylene glycol molecules with long-circulation blood function and protein molecules with tumor cell targeting. The polyethylene glycol modification significantly improves the circulation stability of nanoparticles in the blood, effectively reducing the possibility of recognition and clearance by the immune system, thereby extending their half-life in the body. This not only allows the drug to remain within the effective concentration range for a longer period of time, but also improves the drug's bioavailability. The introduction of protein molecules with tumor cell targeting function, such as antibody fragments and receptor ligands, enables the nanoparticles to specifically recognize and bind to the surface of tumor cells. This high degree of targeting reduces drug exposure to non-target tissues, reduces side effects, and enhances therapeutic efficacy.

[0032] The present invention connects manganese dioxide and near-infrared fluorescent molecules to the surface of the particles through simple chemical modification, giving the system dual-mode imaging capabilities of magnetic resonance imaging and fluorescence imaging. Under the guidance of imaging, the distribution of particles in the organism and the enrichment of lesion areas can be accurately determined, 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 accurately diagnose the disease in real time and simultaneously perform treatment, making it possible to monitor the efficacy during treatment and adjust the dosage regimen at any time, thereby facilitating the optimal treatment effect. Especially for the treatment of diseases such as tumors, this real-time and accurate monitoring and control capability can significantly improve the treatment effect and alleviate patient suffering.

[0033] The present invention adopts a simple and efficient preparation and modification method to prepare hollow nanoparticles with a spiny structure, achieving 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.

[0034] 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 nanoreagent is also determined by the effect of being internalized by tumor cells. The research of the present invention 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. Compared with the same type of particles without thorny structure, the enrichment in tumor cells is increased by about 5-8 times. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] 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 spiny hollow nanoparticles.

[0036] Figure 2 X-ray diffraction patterns of spiny / spineless hollow nanoparticles.

[0037] Figure 3 Thermogravimetric analysis of spiny / spineless hollow nanoparticles.

[0038] Figure 4 The magnetic resonance T1 signal of the spiny / spineless hollow nanoparticles changes over time at pH 5 or 10 mM GSH.

[0039] Figure 5 The graph shows the change of near-infrared fluorescence (810 nm) signal of spiny / spineless hollow nanoparticles at pH 5 or 10 mM GSH over time.

[0040] Figure 6 The graph shows the content of spiny / spineless hollow nanoparticles in tumor cells A549 at different concentrations.

[0041] Figure 7 The spiny / spineless hollow nanoparticle dispersion was exposed to near-infrared light (808 nm, 1 W / cm -2） Variation of dispersion temperature with irradiation time under irradiation.

[0042] 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 a dispersion sample of spiny / spineless hollow nanoparticles.

[0043] Figure 9 Images of mouse tumors and tumor mass were taken 20 days after photothermal therapy with spiny / spineless hollow nanoparticles.

[0044] Figure 10 This figure shows the changes in body weight during photothermal therapy of mouse tumors. DETAILED DESCRIPTION

[0045] 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 with reference to specific embodiments and drawings, but the embodiments are not intended to limit the present invention.

[0046] This invention achieves the controlled preparation of spiny nanoparticles through an innovative chemical reaction pathway, combined with specific reactant and modifier ratios. This method is not only simple to operate and requires mild conditions, but also allows for precise control of particle size and the density and length of the surface spiny structures. The resulting spiny nanoparticles have a uniform size distribution, and the dense surface spiny structures significantly improve photothermal conversion efficiency and cellular endocytosis, demonstrating synergistically enhanced therapeutic effects on various tumor types.

[0047] 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;

[0048] 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.

[0049] 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.

[0050] The spiny carbon nanoparticles are prepared according to the following steps:

[0051] Using silica nanoparticles as core templates, dopamine hydrochloride and tetraethyl silicate are copolymerized on the particle surface to form a shell layer. This is followed by a carbonization process to form a carbon-silicon composite shell layer. The silicon element is then etched away to obtain hollow-structured spiny carbon nanoparticles.

[0052] The mass ratio of dopamine hydrochloride to tetraethyl silicate is 1:5-10.

[0053] During etching, sodium hydroxide solution is used to etch and remove silicon elements.

[0054] The spiny carbon nanoparticles have a particle size of 150-200 nm, and the silicon dioxide has a particle size of 50-120 nm. The spiny structure morphology can be controlled by adjusting the ratio of dopamine hydrochloride and tetraethyl silicate.

[0055] The thickness of the manganese dioxide shell is 20-30 nm, and the loading is 5-15%;

[0056] The modified amount of the fluorescent imaging molecule accounts for 1-2% of the mass of the spiny carbon nanoparticles;

[0057] The modified amount of the targeting ligand molecule accounts for 2-4% of the mass of the spiny carbon nanoparticles.

[0058] Preferably, the thickness of the manganese dioxide shell is 25 nm, and the loading amount is 10%; the modified amount of the fluorescent imaging molecule accounts for 1.5% of the mass of the nanoparticle; and the modified amount of the targeting ligand molecule accounts for 3% of the mass of the nanoparticle.

[0059] The fluorescent imaging molecules include: Cypate, Cyanine7, IR820 and other molecules with emission wavelengths in the near-infrared region.

[0060] The targeting ligand molecules include: transferrin, folic acid, RGD polypeptide and other proteins that can specifically target tumor cells, antibodies, polysaccharides, and P-selectin.

[0061] The particle size of the silicon dioxide nanoparticles is about 100 nm. Silica nanoparticles of different particle sizes can be used as the core to control the particle size of the prepared spiny carbon nanoparticles.

[0062] The manganese dioxide shell deposition process specifically includes adding potassium permanganate to a fully dispersed suspension of spiny nanoparticles, stirring vigorously, and reacting at 80 degrees Celsius for 10 hours to produce spiny hollow nanoparticles of composite manganese dioxide. The mass ratio of potassium permanganate to spiny carbon nanoparticles can be adjusted to adjust the manganese dioxide content on the surface of the spiny nanoparticles, with an optimal ratio of 1:5.

[0063] 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.

[0064] The targeting ligand molecule is a tumor cell-specific targeting molecule, including antibodies, membrane proteins, polypeptides, polysaccharides, etc.

[0065] The manganese dioxide shell is modified with fluorescent molecules and targeting molecules by the following method: first, the spiny 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 attached by the reaction between the amino groups and the activated carboxyl groups, and then the polyethylene glycol molecules are attached to the particle surface, and finally the targeting molecules are added to obtain spiny hollow nanoparticles carrying fluorescent molecules and targeting molecules.

[0066] A second aspect of the present invention provides a method for preparing spiny structure nanoparticles, comprising the following steps:

[0067] Preparation of spiny carbon nanoparticles with hollow structures;

[0068] The spiny carbon nanoparticles are uniformly dispersed in an aqueous 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 deposited on the surface.

[0069] The particles with a manganese dioxide shell deposited on the surface were uniformly dispersed in ethanol, 3-aminopropyltriethoxysilane was added, and the mixture was reacted at 55-65°C for 12-24 hours to obtain particles with amino groups modified on the surface;

[0070] The obtained particles modified with amino groups 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.

[0071] Wherein, the mass ratio of the potassium permanganate to the spiny carbon nanoparticles is 1:3-7.

[0072] The polyethylene glycol molecules include two types, one of which has an NHS ester at one end and a maleimide at the other end, and has a molecular weight of 2000; the other has an NHS ester at one end and a methoxy group at the other end, and has a molecular weight of 2000; the weight ratio of the two polyethylene glycols is 1:4.

[0073] The amount of 3-aminopropyltriethoxysilane added is 10% of the mass of the manganese dioxide particles deposited on the surface.

[0074] A third aspect of the present invention provides a use of spiny structured nanoparticles in the preparation of tumor drugs.

[0075] 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.

[0076] Example 1

[0077] This example is a preparation method and application verification of spiny hollow nanoparticles, which mainly includes the following steps:

[0078] (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.

[0079] (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 the addition of 100 mg of dopamine hydrochloride and 1 mL of tetraethyl silicate. The mixture was reacted 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.

[0080] (3) Preparation of hollow carbon nanoparticles without spiny structures. The steps are basically the same as (2), except that only 200 mg of dopamine hydrochloride is added instead of 100 mg of dopamine hydrochloride and 1 mL of tetraethyl silicate.

[0081] (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°C for 10 hours to obtain nanoparticles with manganese dioxide deposited on the surface, as shown in FIG. Figure 1 Transmission electron microscopy images show that the prepared particles have a particle size of approximately 180 nm. 100 mg of the above particles were dispersed in 100 mL of ethanol, and 50 μL of 3-aminopropyltriethoxysilane was added. The mixture was reacted at 60°C for 24 hours to obtain particles with surface modifications of amino groups. 20 mg of the above particles were dissolved in 20 mL of DMF, and 5 mg of NHS ester-activated Cypate (a near-infrared fluorescent molecule), 4 mg of Mal-PEG-NHS, and 12 mg of mPEG-NHS were added. After stirring at room temperature for 24 hours, PEG- and fluorescent-modified spiny nanoparticles were obtained, namely, spiny-C@MnO2 and spherical-C@MnO2, respectively.

[0082] (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, and the results showed that 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 by magnetic resonance equipment. Figure 4It 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 spicules are similar, indicating that the endocytosis of the two particles can be judged by detecting the strength of the fluorescence signal in the cells.

[0083] (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 5 The cells in each well of the culture plate 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.

[0084] Example 2

[0085] This example describes a method for optimizing spiny nanoparticles and evaluating the efficacy of synergistically enhanced photothermal therapy. 20 mg of the PEG- and fluorescent-modified spiny nanoparticles obtained in Example 1 were mixed with 2 mg of thiolated transferrin at room temperature for 24 hours to obtain spiny nanoparticles with tumor cell targeting capabilities.

[0086] (1) Photothermal performance verification of spiny nanoparticles. The dispersion of spiny / non-spiny nanoparticles with a concentration of 50 μg / mL was placed under the conditions of 808 nm and 1 W / cm -2 The temperature of the dispersion was monitored and recorded in real time using a thermal imager under near-infrared light. Figure 7 As shown, nanoparticles with a spiny structure exhibit better photothermal effects than nanoparticles without a spiny structure, and can raise the temperature of the dispersion to about 60 degrees Celsius within 5 minutes.

[0087] Among them, PBS (physiological saline) is attached Figure 7-10 The group of mice in the control group were injected with PBS via the tail vein. Spiky-C@MnO2-Tf (spiky nanoparticles with fluorescent molecules, PEG and transferrin on the surface) were injected into the adjacent Figure 7-10The experimental group in which the mice were injected with spiny nanoparticles via the tail vein. Figure 7-10 The middle represents the experimental group in which the mice were injected with spikeless nanoparticles through the tail vein.

[0088] (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 ,Use a thermal imager to record the temperature changes of the tumor area in real time, such as Figure 8 As shown in the figure, the temperature of the tumor area of ​​mice injected with nanoparticles with thorny structures reached about 50 degrees Celsius after irradiation, while that of the group without thorny structures was only 30 degrees Celsius. This shows that the thorny structure on the surface of the particles not only increases the amount of enrichment in the tumor area but also has a better photothermal conversion effect, thereby obtaining a higher temperature in the lesion area. Subsequently, a vernier caliper was used to measure the volume of the mouse tumor every other day and record it. On the 20th day, the mice were euthanized and the tumor tissue was removed, photographed, weighed and recorded. Figure 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.

[0089] (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. Figure 10 It can be seen that the weight changes of mice in the experimental group were not significantly different from those in the control group, indicating that the spiny structured nanoparticles have good biosafety.

[0090] The present invention describes preferred embodiments and their effects. However, those skilled in the art, once informed of the basic inventive concept, may make additional changes and modifications to these embodiments. Therefore, it is intended that the appended claims be interpreted to include the preferred embodiments as well as all changes and modifications that fall within the scope of the invention.

[0091] While 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 these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

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

2. The spiny structure nanoparticle according to claim 1, 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.

3. 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 modified amount of the fluorescent imaging molecule accounts for 1-2% of the mass of the spiny carbon nanoparticles; The modified amount of the targeting ligand molecule accounts for 2-4% of the mass of the spiny carbon nanoparticles.

4. The spiny structure nanoparticle according to claim 1, characterized in that The fluorescent imaging molecule is Cypate, a molecule having an emission wavelength in the near-infrared region; The targeting ligand molecule is an antibody, transferrin, folic acid or RGD polypeptide.

5. A method for preparing the spiny structure nanoparticles according to any one of claims 1 to 4, characterized in that: The following steps are involved: Preparation of spiny carbon nanoparticles with hollow structures; The spiny carbon nanoparticles are uniformly dispersed in an aqueous 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 deposited on the surface. The particles with a manganese dioxide shell deposited on the surface were uniformly dispersed in ethanol, 3-aminopropyltriethoxysilane was added, and the mixture was reacted at 55-65°C for 12-24 hours to obtain particles with amino groups modified on the surface; The obtained particles modified with amino groups 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.

6. The method for preparing spiny structure nanoparticles according to claim 5, characterized in that: The mass ratio of the potassium permanganate to the spiny carbon nanoparticles is 1:3-7.

7. The method for preparing spiny structure nanoparticles according to claim 5, characterized in that: The polyethylene glycol molecules include two types, one of which has an NHS ester at one end and a maleimide at the other end, and has a molecular weight of 2000; the other has an NHS ester at one end 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.

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

9. Use of the spiny nanoparticles according to any one of claims 1 to 4 in preparing a drug for photothermal therapy of tumors.

Images