CN114601927A - Monoatomic platinum composite cerium dioxide nano particle for tumor electrokinetic force treatment and preparation thereof - Google Patents

Monoatomic platinum composite cerium dioxide nano particle for tumor electrokinetic force treatment and preparation thereof Download PDF

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CN114601927A
CN114601927A CN202210310010.XA CN202210310010A CN114601927A CN 114601927 A CN114601927 A CN 114601927A CN 202210310010 A CN202210310010 A CN 202210310010A CN 114601927 A CN114601927 A CN 114601927A
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陈莹
尹维翰
谢晨
范曲立
况培培
黄逾东
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Nanjing University of Posts and Telecommunications
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Abstract

The invention provides a monatomic platinum composite cerium dioxide nanoparticle for tumor electrokinetic force treatment and a preparation method thereof. Under the action of an electric field with accurately controllable external strength and position, the composite nano particles can generate Reactive Oxygen Species (ROS) with a tumor killing effect in a tumor, so that the tumor treatment effect is achieved. And because the cerium dioxide has peroxidase-like property, the composite nano particles can catalyze hydrogen peroxide over-expressed in tumor cells to generate hydroxyl free radicals, and the effect of chemical dynamic therapy is achieved. Therefore, the composite nano-particles can be used for chemodynamic and electrodynamic combination therapy, and overcomes the defect of poor activity of the conventional chemodynamic therapy. Moreover, the controllability of the external electric field can effectively improve the selective treatment capability and the space precision of the material, and realize the targeted treatment of the tumor.

Description

Single-atom platinum composite cerium dioxide nano-particle for tumor electrokinetic force treatment and preparation thereof
Technical Field
The invention belongs to the technical field of inorganic nano enzyme, and particularly relates to a monatomic platinum composite cerium dioxide nano particle for tumor electrokinetic force treatment and a preparation method thereof.
Background
In recent years, chemodynamic therapy (CDT), which is a new tumor treatment technology based on the transformation reaction of endogenous chemical products of tumors, has been one of the hot research points in the field of cancer treatment. Unlike conventional tumor treatment techniques, chemokinetic therapy utilizes the tumor microenvironment to activate the fenton reaction (fenton-like reaction) to generate strongly oxidizing hydroxyl radicals for tumor-specific therapy, which utilizes the fenton/fenton-like reaction to induce apoptosis and necrosis. In the Fenton reaction, the transition metal ion catalyzes hydrogen peroxide (H)2O2) The decomposition of the active oxygen (ROS) into hydroxyl radicals (OH-), which are the most harmful active oxygen (ROS), destroys important components (such as lipid, protein or DNA) of tumor cells, and further leads to apoptosis or necrosis, which is the key point of the therapy, however, due to the relatively high pH environment (the pH value in the tumor tissue is 6.5-6.8), endogenous H is generated2O2The concentration of (A) is low, and the expression of reducing substances at the tumor part is high, so that the malignant tumor is difficult to treat by simply using CDT, and the conventional CDT therapy has poor targeting property and selectivity and more limited clinical application.
The electrochemical therapy is a treatment method for achieving a tumor cell killing effect by directly inserting a plurality of electrodes into a tumor and applying direct current to the electrodes and utilizing drastic pH change near the inserted electrodes, and has been clinically applied in some countries. Sharp pH changes near the inserted electrode are a major cause of tumor damage. This method has the advantages of simple operation and small wound, but the limited effective treatment area and the complex electrode configuration thereof also prevent the further development and application of the electrochemical therapy. And no relevant research has been carried out for a long time, which indicates that the electric field has the function of promoting the generation of the nano material (ROS).
After the diligent efforts of the technicians in the field, the caucasian african and asahi et al (Advanced Materials,2019.DOI:10.1002/adma.201806803) have proposed a new conceptual novel way for treating tumors with electromotive force in recent years, and it is shown that platinum nanoparticles can promote the decomposition of water molecules on the surface thereof under the assistance of a direct current electric field and chloride ions to generate ROS with cytotoxicity, which can not only degrade methylene blue, but also effectively inhibit the proliferation of tumor cells to induce apoptosis. This mechanism, known as electrokinetic therapy (EDT), is capable of effectively killing cancer cells throughout the electric field, unlike electrochemotherapy, which is limited to the area near the electrodes. The method is realized mainly by depending on platinum nanoparticles with electrocatalysis function, compared with the traditional method, the EDT method has the advantages of minimal invasion, high selectivity and controllability, strong reaction sustainability, no influence of the surrounding environment of the tumor, and capability of providing uniform killing effect on the whole tumor with relatively large volume.
However, the above research focuses mainly on the catalytic effect of platinum nanoparticles, and it is well known to researchers in the field that the surface effect is very important for heterogeneous catalysis, and the reduction of the size of the active component and the increase of the specific surface of the active component can effectively improve the utilization efficiency of the active component material; the monatomic catalyst may have higher activity relative to nanoparticles because the individual atoms in the monatomic catalyst are all surface atoms; and the monoatomic form in the monoatomic catalyst is single, and high reaction specificity can be formed, so that the monoatomic catalyst can also have high selectivity. Because the prior art does not have relevant research and discussion about the effect of the monatomic platinum in the electrokinetic therapy, the monatomic platinum composite cerium dioxide nanoparticle for the electrokinetic tumor therapy is developed, and the material is expected to be capable of killing tumors with high selectivity and high efficiency in a high pH environment, so that a new idea is provided for the field of tumor therapy.
Disclosure of Invention
The invention aims to overcome the defect of poor activity of conventional chemical dynamic therapy and provides a monatomic platinum composite cerium dioxide nanoparticle for tumor electrokinetic therapy and a preparation method thereof.
The specific technical scheme of the invention is as follows: the preparation method of the monatomic platinum composite cerium dioxide nano-particles for tumor electrokinetic force treatment specifically comprises the following steps:
(1) preparation of cerium hydroxide nanoparticles: dissolving a cerium source in a sodium hydroxide solution, stirring and mixing, then putting into a high-pressure reaction kettle, reacting for several hours at a certain temperature, centrifugally separating out bottom sediment, washing the sediment to be neutral, and then carrying out freeze-drying treatment;
(2)CeO2preparing nano particles: dispersing the freeze-dried sample prepared in the step (1) in a surface modifier under the assistance of ultrasonic waves to connect hydrophilic molecules on cerium hydroxide nanoparticles, separating precipitates after aging treatment, performing acid treatment on the precipitates, washing and drying to prepare CeO2Nanoparticles;
(3) monoatomic platinum composite CeO2Preparing nano particles: the CeO prepared in the step (2)2Mixing the nanometer particles and the platinum source solution, stirring and reacting for several hours at 70 ℃, centrifugally separating out precipitate, washing the precipitate to be neutral, and drying to obtain the product.
Further, in the step (1), the cerium source used is preferably cerium nitrate, cerium acetate or cerium chloride, and other cerium sources capable of achieving the same effect can also be suitable for use herein; the temperature of hydrothermal reaction in the autoclave is 105-170 ℃, the reaction time is 4-24 h, the specific reaction conditions can be determined according to the size of the composite nanoparticles to be obtained, and the ceria particles used as the carrier can be rod-shaped, spherical, sheet-shaped and the like, and are not limited herein.
Further, in the step (2), preferably, the surface modifier is hydroxypropyl cellulose or polyvinylpyrrolidone, and other surface modifiers that achieve equivalent effects may be used.
Further, in the step (2), the temperature during the aging treatment is 70-150 ℃, and the aging treatment time is 8-24 hours.
Further, in the step (2), the molar ratio of the surface modifier to the cerium source is 0.01 to 0.05.
Further, in the step (3), the molar ratio of the platinum source to the cerium source is 0.01 to 0.1.
The monatomic platinum composite cerium dioxide nano-particles prepared by the method can generate Reactive Oxygen Species (ROS) with tumor killing effect in the tumor under the action of an electric field with accurately controllable external strength and position in the environment with the pH value less than 6.8, so the nano-particles have obvious tumor catalytic treatment effect.
Compared with the prior art, the invention has the following advantages:
1. the monatomic platinum composite cerium dioxide nanoparticle overcomes the defect of poor activity of conventional chemical power therapy, and can generate active oxygen (ROS) with a tumor killing effect in the tumor under the action of an electric field with external strength and position capable of being accurately controlled, so that the effect of treating the tumor is achieved. Meanwhile, the material can also show strong catalase-like property, and can realize the effect of CDT and EDT combined treatment;
2. the composite nano-particle prepared by the application takes cerium dioxide as a carrier, and the single-atom platinum is loaded on the cerium dioxide, so that compared with a platinum nano-particle, the single-atom catalyst has larger specific surface area, thereby having higher activity and being capable of improving the EDT treatment effect;
3. because the carrier cerium dioxide is connected with hydrophilic molecules, the nano particles have stronger biocompatibility and can adapt to normal cell physiological conditions;
4. the external electric field can accurately control the strength and the spatial position, effectively improve the selective treatment capability and the spatial precision of the material, realize the maximum protection of normal tissues, carry out targeted treatment on tumors and play a role in uniformly killing the whole tumor with relatively large volume.
Drawings
FIG. 1 is an electron microscope image of the monatomic platinum composite cerium oxide nanoparticles prepared in example III;
FIG. 2 is a high-resolution TEM image of the single-atom Pt/Ce oxide nanoparticles prepared in example III;
FIG. 3 is an absorption diagram of different reaction systems measured under different conditions in example III;
FIG. 4 is a graph of the decomposition rate of the ROS detecting agent DPBF measured in example III as a function of time;
Detailed Description
The technical solution of the present invention is further described below with reference to the accompanying drawings, but not limited thereto, and any modification or equivalent replacement of the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention shall be covered by the protection scope of the present invention.
Example one
(1) Preparation of cerium hydroxide nanoparticles: 0.5g of CeCl3·7H2Dissolving O in 30mL of sodium hydroxide (with the concentration of 3mol/L) solution, stirring at room temperature for 1 hour, putting the obtained solution into a high-pressure reaction kettle, reacting in an oven at 125 ℃ for 12 hours, after the reaction is finished, centrifugally separating out bottom precipitate, washing the precipitate with ultrapure water to be neutral, and then carrying out freeze-drying treatment;
(2) cerium oxide (CeO)2) Preparing nano particles: dispersing 2mg of the freeze-dried powder sample prepared in the previous step in a hydroxypropyl cellulose (1mg/mL) aqueous solution under the assistance of ultrasound, carrying out water bath heating aging treatment for 12 hours at 80 ℃, centrifugally separating out light yellow precipitate, washing the precipitate twice with a hydrochloric acid solution (1mol/L), then alternately washing with ethanol and water for three times, drying at 60 ℃, and finally obtaining white powdered CeO2Nanoparticles;
(3) preparation of composite nanoparticles loaded with monatomic platinum: dispersing the white powder obtained in the previous step into chloroplatinic acid (1mg/mL) aqueous solution according to the concentration of 1mg/mL, stirring for 12 hours at 70 ℃, centrifuging (the rotating speed is more than 6000 r/min), washing the precipitate obtained twice with ultrapure water to be neutral, and drying at 70 ℃ to obtain the product.
Example two
(1) Preparation of cerium hydroxide nanoparticles: 0.5g of CeCl3·7H2Dissolving O in 30mL of sodium hydroxide (with the concentration of 3mol/L) solution, stirring at room temperature for 1 hour, putting the obtained solution into a high-pressure reaction kettle, reacting in an oven at 125 ℃ for 12 hours, after the reaction is finished, centrifugally separating out bottom precipitate, washing the precipitate with ultrapure water to be neutral, and then carrying out freeze-drying treatment;
(2) cerium oxide (CeO)2) Preparing nano particles: dispersing 2mg of the freeze-dried powder sample prepared in the previous step in a polyvinylpyrrolidone (PVP, 1mg/mL) aqueous solution under the assistance of ultrasound, carrying out water bath heating aging treatment for 12 hours at 80 ℃, centrifuging to separate a light yellow precipitate, washing the precipitate twice with a hydrochloric acid solution (1mol/L), then alternately washing with ethanol and water for three times, drying at 60 ℃, and finally obtaining white powdered CeO2Nanoparticles;
(3) preparation of composite nanoparticles loaded with monatomic platinum: dispersing the white powder obtained in the previous step into a potassium chloroplatinate (1mg/mL) aqueous solution according to the concentration of 1mg/mL, stirring for 12 hours at 70 ℃, centrifuging (the rotating speed is more than 6000 r/min), washing the bottom precipitate twice with ultrapure water to be neutral, and drying at 70 ℃ to obtain the product.
EXAMPLE III
(1) Preparation of cerium hydroxide nanoparticles: 0.56g of Ce (NO)3)3Dissolving in 30mL of sodium hydroxide (with the concentration of 3mol/L) solution, stirring at room temperature for 1 hour, putting the obtained solution into a high-pressure reaction kettle, reacting in an oven at 125 ℃ for 12 hours, after the reaction is finished, centrifugally separating out bottom precipitate, washing the precipitate with ultrapure water to be neutral, and then carrying out freeze-drying treatment;
(2) cerium oxide (CeO)2) Preparing nano particles: dispersing 2mg of the lyophilized powder sample prepared in the previous step in a hydroxypropyl cellulose (1mg/mL) aqueous solution under the assistance of ultrasound, carrying out heating and aging treatment in a water bath at 80 ℃ for 12 hours, centrifugally separating out light yellow precipitate, washing the precipitate twice with a hydrochloric acid solution (1mol/L), then washing the precipitate three times with ethanol and water alternately, and drying the precipitate at 60 ℃ to finally obtain white powdered CeO2Nanoparticles;
(3) preparation of composite nanoparticles loaded with monatomic platinum: dispersing the white powder obtained in the previous step into chloroplatinic acid (1mg/mL) aqueous solution according to the concentration of 1mg/mL, stirring for 12 hours at 70 ℃, centrifuging (the rotating speed is more than 6000 r/min), washing the bottom precipitate twice with ultrapure water to be neutral, and drying at 70 ℃ to obtain the product.
And (4) relevant performance test:
1. preparing a sample of the material, wherein a transmission electron microscope image of the sample is shown in FIG. 1; FIG. 2 is taken under a high-resolution transmission electron microscope, and it can be seen from FIG. 2 that platinum atoms are present as single atoms on the carrier.
2. Preparing the prepared final product into an aqueous dispersion solution with the concentration of 1mg/mL, and adding quantitative hydrogen peroxide and a color developing agent 3,3',5,5' -Tetramethylbenzidine (TMB) into the solution; after the mixed solution reacts for 30min, a spectrophotometer is used for measuring the absorbance-wavelength curve of the system, the relevant result is shown in figure 3, and the TMB + H and the control group can be seen from figure 32O2Compared with the nanometer particles + TMB + H without the action of the electric field2O2The group absorbance was significantly greater, demonstrating the strong peroxidase-like nature of the material. And the electric field action + nanoparticles + TMB + H2O2The absorbance of the group is the maximum, and the fact that the material can generate ROS to oxidize TMB under the action of an electric field is proved.
3. The prepared final product was prepared as an aqueous dispersion solution with a concentration of 1mg/mL, a buffer and ROS detection agent 1, 3-Diphenylisobenzofuran (DPBF) were added to the solution, the developer reacted with the generated ROS to generate colorless substances, electricity was applied for a certain time after mixing, and the DPBF concentration was converted by measuring the system absorbance using a spectrophotometer to obtain the DPBF decomposition rate, and as a result, as shown in fig. 4, the control group was added with DPBF only in an aqueous dispersion of the same amount of nanoparticles without the application of an electric field, and as shown in fig. 4, when no electric field was applied, the ROS detection agent DPBF was not decomposed substantially when coexisted with the nanoparticles. The decomposition rate of DPBF of the system is increased along with time under the action of an electric field, and the fact that the material can generate ROS under the action of the electric field is proved.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes performed by the present specification and drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims (7)

1. The preparation method of the monatomic platinum composite cerium dioxide nano-particles for tumor electrokinetic force treatment is characterized by comprising the following steps:
(1) preparation of cerium hydroxide nanoparticles: dissolving a cerium source in a sodium hydroxide solution, stirring and mixing, then putting into a high-pressure reaction kettle, reacting for several hours at a certain temperature, centrifugally separating out bottom sediment, washing the sediment to be neutral, and then carrying out freeze-drying treatment;
(2)CeO2preparing nano particles: dispersing the freeze-dried sample prepared in the step (1) in a surface modifier under the assistance of ultrasonic waves to connect hydrophilic molecules on cerium hydroxide nanoparticles, separating precipitates after aging treatment, performing acid treatment on the precipitates, washing and drying to prepare CeO2Nanoparticles;
(3) monoatomic platinum composite CeO2Preparing nano particles: the CeO prepared in the step (2)2Mixing the nanometer particles and the platinum source solution, stirring and reacting for several hours at 70 ℃, centrifugally separating out precipitate, washing the precipitate to be neutral, and drying to obtain the product.
2. The method for preparing the monatomic platinum composite cerium oxide nanoparticles for electrodynamic oncotherapy according to claim 1, wherein in the step (1), the cerium source used is cerium nitrate, cerium acetate or cerium chloride; the temperature of hydrothermal reaction in the autoclave is 105-170 ℃, and the reaction time is 4-24 h.
3. The method for preparing the monatomic platinum composite ceria nanoparticles for electrokinetic tumor therapy according to claim 1, wherein in the step (2), the surface modifier is hydroxypropylcellulose or polyvinylpyrrolidone.
4. The method for preparing the monatomic platinum composite ceria nanoparticles for electrokinetic tumor therapy according to claim 1, wherein the aging treatment in the step (2) is performed at a temperature of 70 to 150 ℃ for 8 to 24 hours.
5. The method of preparing the monatomic platinum composite ceria nanoparticles for electrokinetic tumor therapy according to claim 1, wherein in the step (2), the molar ratio of the surface modifier to the cerium source is 0.01 to 0.05.
6. The method for preparing the monatomic platinum composite cerium oxide nanoparticles for electrokinetic tumor therapy according to claim 1, wherein in the step (3), the molar ratio of the platinum source to the cerium source is 0.01 to 0.1.
7. The nanoparticles prepared by the preparation method of the monatomic platinum composite cerium dioxide nanoparticles for electrokinetic tumor therapy according to any one of claims 1 to 6, wherein the nanoparticles catalyze the generation of ROS in an aqueous solution under the action of an external electric field in an environment with pH < 6.8.
CN202210310010.XA 2022-03-28 2022-03-28 Monoatomic platinum composite cerium dioxide nano particle for tumor electrokinetic force treatment and preparation thereof Pending CN114601927A (en)

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Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HUIJING XIANG等: "Single-Atom Catalysts in Catalytic Biomedicine", ADVANCED MATERIALS *
TONGXU GU等、: "Platinum Nanoparticles to Enable Electrodynamic Therapy for Effective Cancer Treatment", ADVANCED MATERIALS *
张玉涛: "氧化铈基纳米酶的制备及其在肿瘤治疗方面的应用研究", 中国优秀硕士学位论文全文数据库医药卫生科技辑 *

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Application publication date: 20220610