Preparation method and application of cuprous selenide photo-thermal chemical power nanometer platform loaded with calcium peroxide

Abstract

The invention belongs to the field of nano material preparation, and discloses a preparation method and application of a cuprous selenide photo-thermal chemical power nano platform loaded with calcium peroxide, and a nano compound is prepared from cuprous selenide nano particles, calcium peroxide nano particles carrying hyaluronic acid and an organic phase change material. The surfaces of the cuprous selenide nano-particles are loaded with the hyaluronic acid calcium peroxide nano-particles, and the outer layers of the cuprous selenide nano-particles are coated with the organic phase change material. The preparation method of the nano-composite is simple, large-scale production is easy, a new preparation mode is adopted, the prepared Cu2Se structure is more stable and can be conveniently combined with subsequent calcium peroxide, and the quality and the using effect of a nano-platform are improved; meanwhile, the preparation mode of calcium peroxide is improved, combined operation steps are simplified, the used raw materials, the feeding proportion, the feeding sequence and the reaction temperature are limited and optimized, the preparation efficiency and quality of the nano platform are greatly improved, and large-scale production and use of medical materials are facilitated.

Description

Technical Field

[0001] The invention relates to the field of nano material preparation, and in particular to a preparation method and application of a cuprous selenide photothermochemical power nano platform loaded with calcium peroxide. Background Art

[0002] At present, the treatment of cancer patients is mainly based on a comprehensive treatment of radical resection combined with postoperative adjuvant chemotherapy. However, traditional chemotherapy drugs (such as oxaliplatin, paclitaxel, etc.) are prone to cause systemic multi-system toxic side effects such as hair loss, nausea and vomiting, and bone marrow suppression, and are also prone to drug resistance, which reduces the anti-tumor effect of chemotherapy drugs to a certain extent. In addition to traditional chemotherapy, more and more new tumor prevention and treatment methods are being used in tumor treatment. Among them, chemodynamic therapy (CDT) is one of the more effective emerging cancer treatment strategies. The so-called chemodynamic therapy (CDT) refers to a new treatment mode in which endogenous hydrogen peroxide (H2O2) in cells undergoes Fenton or Fenton-like reactions in the presence of metal catalysts under the acidic microenvironment of the tumor and produces a large number of hydroxyl free radicals (·OH), which can then oxidatively damage lipids, proteins, and DNA, causing cancer cell death. So far, iron-based nanomaterials, such as FeS2, Fe2P, and Fe5C2@MnO2, are the main drugs used for CDT to kill tumors. However, the Fenton effect based on iron materials is only effective under strong acidic conditions (pH = 3-5), while the Fenton reaction efficiency is not high under weakly acidic conditions (pH = 6.8) in tumor cells. In addition, even at a suitable pH value, the Fenton reaction efficiency of iron-based nanomaterials is relatively low, resulting in slow generation of ·OH. Therefore, how to improve the efficiency of the Fenton reaction at the tumor site will be the key to further improving the efficacy of CDT.

[0003] In recent years, photothermal therapy (PTT) has attracted more and more attention. It is an emerging tumor treatment method that uses photosensitizers to convert near-infrared light energy into heat energy to destroy cancer cells. It has the advantages of wide application range, non-invasiveness, strong selectivity, simple process, and little damage to normal tissues. It has shown great application value in the field of tumor treatment. Interestingly, studies have found that photothermal therapy (PTT) can increase the local temperature of the tumor site, and a slight increase in temperature can significantly enhance the Fenton or Fenton-like effect. For example, FePS3 nanosheets are efficient photothermal agents with Fenton catalytic activity. The Fenton catalytic effect can be significantly enhanced under photothermal action, but it is still a material based on iron, so the Fenton effect enhancement is limited. At present, the nanomaterials commonly used in photothermal therapy, such as carbon nanorods, graphene oxide (GO), gold nanorods (Au), copper sulfide (CuS), etc., are mainly excited by near-infrared region I (NIRI) lasers to generate heat under the action of alternating magnetic fields to treat tumors. Compared with near-infrared zone 1 nanomaterials, near-infrared zone 2 nanomaterials have two obvious advantages: on the one hand, they have a stronger tissue penetration depth; on the other hand, they allow for higher irradiation doses (according to the national standard for safe use of lasers in the United States, the power density of 1064nm wavelength lasers that the skin can be safely exposed to is 1W / cm2, while for 808nm lasers, this value is only 0.33W / cm2). In order to increase the concentration of hydrogen peroxide in tumor cells and enhance the efficacy of chemodynamics, a related patent has successfully constructed ultra-small calcium peroxide (CaO2), which can react with water to generate hydrogen peroxide on the one hand; on the other hand, it can generate calcium ions in an acidic environment. The enrichment of calcium ions in cells can cause calcium overload in tumor cells and secondary cell apoptosis. However, CaO2 has poor stability and is easily decomposed in water. At present, there is no material that can carry CaO2 and can be excited by near-infrared zone 2 lasers to generate heat under the action of an alternating magnetic field to treat tumors. Summary of the invention

[0004] The present invention is intended to provide a method for preparing a cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide to solve the problems existing in the above-mentioned background technology.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] A method for preparing a cuprous selenide photothermochemical power nano-platform loaded with calcium peroxide, the preparation method comprising the following steps:

[0007] S1. Preparation of cuprous oxide:

[0008] Prepare a first reaction container, add deionized water into the first reaction container, then add copper chloride dihydrate and polyvinyl pyrrolidone, stir for 1h to 2h, then add sodium hydroxide solution, stir again to dissolve the sodium hydroxide solution, introduce nitrogen into the first reaction container so that the first reaction container is in a nitrogen environment, then slowly add ascorbic acid, stir again for 1h to 2h, centrifuge the reactants, purify and dry the obtained solid to obtain cuprous oxide;

[0009] S2. Preparation of cuprous selenide:

[0010] Prepare a second reaction container, add deionized water into the second reaction container, introduce nitrogen into the second reaction container so that the second reaction container is in a nitrogen environment, add selenium powder and sodium borohydride, stir for 23h to 25h, then add cuprous oxide, stir for a second time, centrifuge the reactants after the reaction is completed, purify and dry the obtained solid to obtain cuprous selenide;

[0011] S3, modified cuprous selenide:

[0012] Prepare a third reaction container, add deionized water into the third reaction container, then add cuprous selenide and SH-PEG, stir, centrifuge the reactants after the reaction is completed, purify and dry the obtained solid to obtain modified cuprous selenide (SH-PEG-Cu2Se);

[0013] S4, preparation of calcium peroxide:

[0014] Prepare a fourth reaction container, add hyaluronic acid into the fourth reaction container, stir the hyaluronic acid, add calcium chloride solution while stirring the hyaluronic acid, continue stirring for 23h to 25h, then add ammonia water and hydrogen peroxide in sequence, wait for the reaction for 2h to 4h, then place the reaction system in an ultrasonic environment, add sodium hydroxide to obtain a white emulsion containing HA-CaO2 nanoparticles, wash the corresponding HA-CaO2 nanoparticles with sodium hydroxide, water and ethanol in sequence, and then store the HA-CaO2 nanoparticles in ethanol at 2°C to 6°C;

[0015] S5. Preparation of cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide:

[0016] Prepare a fifth reaction container, add ethanol into the fifth reaction container, then add modified cuprous selenide and HA-CaO2, stir for 23h to 25h, prepare a dissolution container, add ethanol into the dissolution container, then add lauric acid into the dissolution container, dissolve lauric acid in the ethanol in the dissolution container, then add the lauric acid ethanol mixture into the reaction container, stir for a second time for 4h to 6h, after the reaction is completed, separate the reactants to obtain a precipitate, wash the precipitate with ethanol for 2 to 4 times, and you can get a cuprous selenide photothermal chemical power nanocomposite loaded with calcium peroxide.

[0017] Preferably, in step S1, the mass ratio of cupric chloride dihydrate, polyvinyl pyrrolidine and sodium hydroxide is 80-90:30:30-34.

[0018] Preferably, in step S2, the mass ratio of selenium powder to sodium borohydride is 1 to 2:1.

[0019] Preferably, in step S3, the mass ratio of cuprous selenide to SH-PEG is 1 to 2:1.

[0020] Preferably, in step S4, the concentration of the calcium chloride solution is 0.1 g / mL, the concentration of the hyaluronic acid solution is 15 mg / mL, and the volume ratio of the calcium chloride solution to the hyaluronic acid solution is 1:4-6.

[0021] Preferably, in step S5, the mass ratio of modified cuprous selenide, HA-CaO2 and lauric acid is 1:0.8-1.2:98-102.

[0022] Preferably, in step S4, aqueous ammonia is added dropwise to make the pH value of the reaction system be 8.5-10.

[0023] The present invention also provides the use of the cuprous selenide photothermochemical power nanocomposite loaded with calcium peroxide as described in any of the above schemes for tumor treatment.

[0024] The beneficial effects of this technical solution are:

[0025] (1) The present invention adopts a new preparation method to prepare a cuprous selenide nanocomposite as a copper-based nanomaterial. The cuprous selenide nanocomposite has better photothermal properties and is easy to modify. It can be used in the near infrared light region (NIR II) emits light and generates heat under excitation, thereby enhancing its Fenton-like effect in cells; a new preparation method is adopted, Cu2O is used as a template, and an ion replacement method is used to synthesize Cu2Se with a hollow structure. The prepared Cu2Se structure is more stable and is also convenient for combining with subsequent calcium peroxide, thereby improving the quality and use effect of the nano-platform; at the same time, the present application improves the preparation method of calcium peroxide, and reacts in a hyaluronic acid system to generate HA-CaO2 nanoparticles. Under the action of hyaluronic acid, the particle size of the nanocomposite can be controlled to form nanoparticles with ultra-small particle size, thereby improving the preparation efficiency and quality of HA-CaO2 nanoparticles; at the same time, the calcium peroxide nanoparticles interact with hyaluronic acid and cuprous selenide, so that calcium peroxide can be stably adsorbed on cuprous selenide, which greatly simplifies the operation steps of combining the two and also improves the stability of the combination of the two; the preparation efficiency and quality of a cuprous selenide photothermal chemical power nano-platform loaded with calcium peroxide are greatly improved by improving the reaction raw materials and optimizing the preparation method.

[0026] (2) The surface of the nano-platform of the present invention is coated with lauric acid, which makes the nano-platform have a thermosensitive effect. When the temperature rises to a specific temperature range under the action of photothermal, the lauric acid mold of the nano-particles is broken, and the nano-particles are released at a fixed point at the tumor site. The melting point of lauric acid is 42°C to 45°C, which is adapted to the optimal treatment environment temperature, and can improve the use effect of the nano-platform.

[0027] (3) The present invention has made limited optimization on the raw materials used, the proportion of the raw materials, the order of adding the materials, and the reaction temperature, which greatly improves the efficiency and quality of the nano-platform preparation. The raw materials used are low in cost and the reaction conditions are mild, which is convenient for the large-scale production of medical materials.

[0028] (4) Under the irradiation of near infrared light of 1064 nm in the near infrared II region (NIR II), the nanocomplex of the present invention can produce a synergistic effect with the chemical dynamic effect to generate a large amount of ROS, damage mitochondria, and improve the therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Physical and chemical characterization diagram of cuprous selenide and calcium peroxide nanomaterials provided by the present invention;

[0030] Figure 2 A schematic diagram of the chemical kinetic effect of cuprous selenide provided by the present invention;

[0031] Figure 3 A schematic diagram of the ability of calcium peroxide provided by the present invention to produce hydrogen peroxide under different conditions;

[0032] Figure 4 This is a diagram of the anti-tumor effect of Cu2Se-CaO2@LA provided by the present invention;

[0033] Figure 5 This is a diagram showing the damage to mitochondrial function of Cu2Se-CaO2@LA provided by the present invention; DETAILED DESCRIPTION

[0034] The applicant will further explain the present invention in detail below in conjunction with specific examples, in order to enable those skilled in the art to more clearly understand the present invention, but the following content should not be understood in any way as limiting the scope of protection claimed in the claims of this application. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used can all be obtained from commercial channels;

[0035] Embodiment 1:

[0036] The method for preparing a cuprous selenide photothermochemical power nano-platform loaded with calcium peroxide in this embodiment comprises the following steps:

[0037] S1. Preparation of cuprous oxide:

[0038] Prepare a first reaction container, add 20 mL of deionized water into the first reaction container, then add 85 mg of cupric chloride dihydrate and 300 mg of polyvinyl pyrrolidone (MW=6000), stir for 1 hour, then add 320 mg of sodium hydroxide solution, stir again to dissolve the sodium hydroxide solution, introduce nitrogen into the first reaction container so that the first reaction container is in a nitrogen environment, then slowly add ascorbic acid, stir again for 1 hour, react to generate reactants, centrifuge the reactants, purify and dry the obtained solid, and obtain cuprous oxide (Cu2O);

[0039] S2. Preparation of cuprous selenide:

[0040] Prepare a second reaction container, add 20 mL of deionized water into the second reaction container, introduce nitrogen into the second reaction container so that the second reaction container is in a nitrogen environment, add 50 mg of selenium powder and 50 mg of sodium borohydride into the second reaction container, stir for 24 hours, then add cuprous oxide, the cuprous oxide added here is obtained in step S1, stir twice, centrifuge the reactants after the reaction is completed, purify and dry the obtained solid to obtain cuprous selenide;

[0041] S3, modified cuprous selenide:

[0042] Prepare a third reaction container, add 20 mL of deionized water into the third reaction container, then add 10 mg of cuprous selenide and 10 mg of SH-PEG, stir for 24 h, and after the reaction is completed, centrifuge the reactants, purify and dry the obtained solid to obtain modified cuprous selenide (SH-PEG-Cu2Se);

[0043] S4, preparation of calcium peroxide:

[0044] Prepare a fourth reaction container, add 10 mL of a hyaluronic acid solution with a concentration of 15 mg / mL into the fourth reaction container, stir the hyaluronic acid, add 2 mL of a calcium chloride solution with a concentration of 0.1 g / mL while stirring the hyaluronic acid solution, continue stirring for 24 hours, then add 1 mL of ammonia water (1M) and 1.5 mL of hydrogen peroxide (30 wt%) in sequence, add ammonia water dropwise to make the pH value of the reaction system 8.5, wait for the reaction for 3 hours, then place the reaction system in an ultrasonic environment, add 0.5 mL of sodium hydroxide (1M), and obtain a white emulsion containing HA-CaO2 nanoparticles, wash the corresponding HA-CaO2 nanoparticles with sodium hydroxide, water and ethanol in sequence, and then store the HA-CaO2 nanoparticles in ethanol at 4°C;

[0045] S5. Preparation of cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide:

[0046] Prepare a fifth reaction container, add 5 mL of ethanol into the fifth reaction container, then add 1 mg of modified cuprous selenide and HA-CaO2, and stir for 24 hours at a time, prepare a dissolution container, add 1 mL of ethanol into the dissolution container, and then add 0.1 g of lauric acid into the dissolution container to dissolve lauric acid in the ethanol in the dissolution container, then add the lauric acid ethanol mixture into the fifth reaction container, stir for a second time for 5 hours, and after the reaction is completed, separate the reactants to obtain a precipitate, wash the precipitate with ethanol 3 times to obtain a cuprous selenide photothermal chemical power nanoplatform loaded with calcium peroxide (Cu2Se-CaO2@LA).

[0047] Embodiment 2:

[0048] The method for preparing a cuprous selenide photothermochemical power nano-platform loaded with calcium peroxide in this embodiment comprises the following steps:

[0049] S1. Preparation of cuprous oxide:

[0050] Prepare a first reaction container, add 20 mL of deionized water into the first reaction container, then add 85 mg of cupric chloride dihydrate and 300 mg of polyvinyl pyrrolidone (MW=6000), stir for 1 hour, then add 320 mg of sodium hydroxide solution, stir again to dissolve the sodium hydroxide solution, introduce nitrogen into the first reaction container so that the first reaction container is in a nitrogen environment, then slowly add ascorbic acid, stir again for 1 hour, react to generate reactants, centrifuge the reactants, purify and dry the obtained solid, and obtain cuprous oxide (Cu2O);

[0051] S2. Preparation of cuprous selenide:

[0052] Prepare a second reaction container, add 20 mL of deionized water into the second reaction container, introduce nitrogen into the second reaction container so that the second reaction container is in a nitrogen environment, add 50 mg of selenium powder and 50 mg of sodium borohydride into the second reaction container, stir for 24 hours, then add cuprous oxide, the cuprous oxide added here is obtained in step S1, stir twice, centrifuge the reactants after the reaction is completed, purify and dry the obtained solid to obtain cuprous selenide;

[0053] S3, modified cuprous selenide:

[0054] Prepare a third reaction container, add 20 mL of deionized water into the third reaction container, then add 10 mg of cuprous selenide and 10 mg of SH-PEG, stir for 24 h, and after the reaction is completed, centrifuge the reactants, purify and dry the obtained solid to obtain modified cuprous selenide (SH-PEG-Cu2Se);

[0055] S4, preparation of calcium peroxide:

[0056] Prepare a fourth reaction container, add 10 mL of a hyaluronic acid solution with a concentration of 15 mg / mL into the fourth reaction container, stir the hyaluronic acid, add 2 mL of a calcium chloride solution with a concentration of 0.1 g / mL while stirring the hyaluronic acid solution, continue stirring for 24 hours, then add 1 mL of ammonia water (1M) and 1.5 mL of hydrogen peroxide (30 wt%) in sequence, add ammonia water dropwise to make the pH value of the reaction system 8.5, wait for the reaction for 3 hours, then place the reaction system in an ultrasonic environment, add 0.5 mL of sodium hydroxide (1M), and obtain a white emulsion containing HA-CaO2 nanoparticles, wash the corresponding HA-CaO2 nanoparticles with sodium hydroxide, water and ethanol in sequence, and then store the HA-CaO2 nanoparticles in ethanol at 4°C;

[0057] S5. Preparation of cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide:

[0058] Prepare a fifth reaction container, add 5 mL of ethanol into the fifth reaction container, then add 1 mg of modified cuprous selenide and HA-CaO2, stir for 24 hours at a time, and after the reaction is completed, separate the reactants to obtain a precipitate, wash the precipitate with ethanol three times to obtain a cuprous selenide photothermochemical power nanocomposite (Cu2Se-CaO2) loaded with calcium peroxide and not coated with lauric acid.

[0059] Test Example 1: Physical and Chemical Characterization

[0060] 1. Observe the external morphology of the prepared Cu2Se and CaO2 by transmission electron microscopy (TEM) scanning experiment. First, collect and appropriately dilute the sample solution, and then use a pipette to draw 10uL of the sample and drop it on the nickel mesh. After the solution evaporates at room temperature, use an ion sputtering instrument to plate a conductive gold film, and use TEM to observe the morphology of the nanocomposite at an accelerating voltage of 200kV.

[0061] 2. The prepared Cu2Se-CaO2@LA sample was freeze-dried, then ground into powder, mixed with potassium bromide (KBr) and pressed into a granular mixture, and finally analyzed using a Nicolet 7000-C FTIR analyzer to obtain an infrared spectrum of the nanocomposite.

[0062] 3. The prepared Cu2Se-CaO2@LA was freeze-dried and then ground into powder. The X-ray diffraction pattern was then recorded and observed on a PANalyticalX'Pert PRO X-ray diffractometer. At the same time, the X-ray photoelectron spectrum of the nanosample was observed and recorded using a PHI5000VersaProbe spectrometer.

[0063] The above test results can be found in Figure 1 Among them, Figure A from left to right is the transmission electron microscope image of cuprous selenide; transmission electron microscope grayscale image of cuprous selenide, copper element analysis in cuprous selenide electron microscope mapping; selenium element analysis in cuprous selenide electron microscope mapping; Figure B is the XPS analysis of cuprous selenide and calcium peroxide; Figure C is the XRD analysis of cuprous selenide and calcium peroxide; Figure D is the Fourier infrared analysis of cuprous selenide and calcium peroxide.

[0064] Analysis of results: Physical and chemical properties such as infrared absorption spectrum, scanning electron microscopy, X-ray fluorescence diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) all indicate that cuprous selenide and calcium peroxide nanomaterials have been successfully synthesized; cuprous selenide contains copper and selenium elements, in which the copper element is in the cuprous (Cu+) valence state, while calcium peroxide contains calcium and oxygen elements, and no other impurities are found in the two substances.

[0065] Experimental Example 2: Detection of Enhanced Fenton Effect

[0066] Classical colorimetric analysis of ·OH generation based on the degradation of methylene blue (MB) after selective ·OH capture.

[0067] 1: PBS (pH = 6.5); 4mM H2O2; MB solution (10μg / mL), at 25°C, different concentrations of Cu2Se (0μg / mL; 20μg / mL; 40μg / mL; 60μg / mL; 80μg / mL; 100μg / mL) were added respectively;

[0068] 2. PBS (pH = 6.5); 4mM H2O2; MB solution (10μg / mL), at 25℃, add different concentrations of Cu2 Se@CaO2 (0μg / mL; 20μg / mL; 40μg / mL; 60μg / mL; 80μg / mL; 100μg / mL); detect its absorbance at λ = 664nm. Then raise the temperature to 45℃ to simulate the heat during photothermal therapy and evaluate the effect of temperature on ·OH generation. The results are shown in Figure 2 A is the MB degradation curve of Cu2 Se at different concentrations, and the curves from top to bottom correspond to 0μg / mL; 20μg / mL; 40μg / mL; 60μg / mL; 80μg / mL; 100μg / mL of Cu2 Se; B is the MB degradation curve of Cu2 Se@CaO2 at different concentrations, and the curves from top to bottom correspond to 0μg / mL; 20μg / mL; 40μg / mL; 60μg / mL; 80μg / mL; 100μg / mL of Cu2 Se@CaO2; C is the quantitative analysis of A and B at a wavelength of 664nm. Under the same concentration conditions, the left bar graph corresponds to Cu2Se, and the right bar graph corresponds to Cu2Se@CaO2 (i.e., Cu2 Se-CaO2). *** represents P<0.001.

[0069] Result analysis: From Figure 2 From A, we can see that Cu2Se can degrade MB, proving that Cu2Se produces a Fenton effect. At the same time, from B, we can see that Cu2Se@CaO2 produces a stronger Fenton effect. From C, we can see that after quantitative analysis, Cu2Se@CaO2 has a stronger ability to produce a Fenton effect than Cu2Se.

[0070] Test Example 3: Hydrogen Peroxide Generation Detection

[0071] The standard curve was obtained by measuring hydrogen peroxide with potassium permanganate, and the standard curve can be used to quantitatively analyze the concentration of hydrogen peroxide in the solution. First, potassium permanganate (1 mg / mL) was titrated with different concentrations of hydrogen peroxide (0 mM; 20 mM; 40 mM; 60 mM; 80 mM; 100 mM; 200 mM) to obtain a standard curve. According to the standard curve, the corresponding hydrogen peroxide content was calculated. CaO2 was divided into three groups (n=3): (i) dispersed in deionized water (pH=7.0, 100 μg / mL) at 37°C or 50°C. (ii) CaO2 nanoparticles dispersed in PBS buffer (pH=6.5, 100 μg / mL) at 37°C or 50°C. (iii) CaO2 nanoparticles dispersed in PBS buffer (pH=6.0, 100 μg / mL) at 37°C or 50°C. 20 μL of KI (0.1 M) aqueous solution was added to each group. Measure the UV-visible absorption spectrum of the solution and calculate the concentration of generated H2O2 after 5 minutes. The results are shown in Figure 3 , Figure 3 A is the UV absorption graph of potassium permanganate and different concentrations of hydrogen peroxide. After different concentrations of hydrogen peroxide react with potassium permanganate, the UV absorption will change due to the decomposition of potassium permanganate. Therefore, by measuring the UV absorption spectrum after the reaction, the corresponding hydrogen peroxide concentration can be obtained. The curves correspond to the UV absorption spectra of 0mM, 20mM, 40mM, 60mM, 80mM, 100mM, and 200mM hydrogen peroxide titration potassium permanganate from top to bottom; B is the ability of CaO2 to produce hydrogen peroxide under different conditions. Under the same pH conditions, the left bar graph corresponds to 50°C, and the right bar graph corresponds to 37°C.

[0072] Result analysis: From Figure 3 It can be seen that CaO2 has a good ability to produce hydrogen peroxide. At different pH and temperatures, calcium peroxide exhibits good temperature and pH response characteristics.

[0073] Experimental Example 4: Antitumor effect test of Cu2Se@CaO2@LA nanocomposite

[0074] In the CCK-8 cytotoxicity experiment, HGC-27 cells in the logarithmic growth phase were first digested with trypsin to obtain a single cell suspension, and 5000 cells / well were inoculated on a 96-well plate and cultured in a constant temperature incubator for 24 hours; then, after the cells adhered to the wall, different concentrations of Cu2 Se and Cu2 Se-CaO2@LA nanocomplexes were added to each microwell and treated with light or no light, and then continued to be cultured in the incubator for 24 hours; then, the culture medium was removed, and 100 μL of pre-prepared 10% CCK-8 reagent was added to each microwell to be tested; finally, the absorbance of each measuring well at 450 nm was measured using an ELISA reader, and the results were recorded for comparative analysis. The results are shown in Figure 4 .

[0075] Result analysis: From Figure 4 It can be seen that under the action of Cu2Se alone, a certain anti-tumor effect can be produced, and the anti-tumor effect is further increased after NIR II (1064nm) light irradiation; after loading ultra-small CaO2 nanoparticles, the anti-tumor effect is significantly enhanced. Under the protection of the organic phase change material lauric acid LA, no cytotoxic effect is produced.

[0076] Experimental Example 5: Cu2Se@CaO2@LA nanocomplex induced mitochondrial dysfunction test

[0077] Tumor cells were inoculated in 6-well plates and treated with Cu2Se@CaO2@LA nanocomplexes and Cu2Se nanoparticles, respectively. Then, the cells were collected and added with JC-1 detection kit, ROS detection kit, and mPTP detection kit to measure the intracellular mitochondrial membrane potential (MTP), the level of ROS generation, and the degree of opening of mPTP. The results are shown in Table 1. Figure 5 , A is the detection of ROS generation level; B is the detection of intracellular mPTP opening degree; C is the detection of intracellular mitochondrial membrane potential (MTP); *** represents P < 0.001.

[0078] Results: Under the action of Cu2Se, cells can produce reactive oxygen species (ROS) and mitochondrial dysfunction, such as opening of mPTP pore channels and decreased membrane potential; after NIR II (1064nm) light irradiation, the amount of reactive oxygen species produced further increased; after loading ultra-small CaO2 nanoparticles, the amount of ROS production increased significantly. Under the non-light and organic phase change material lauric acid LA restriction, the effect of cells producing ROS was weak.

[0079] The above is only an embodiment of the present invention, and the common knowledge such as the known specific technical solutions and / or characteristics in the solution is not described in detail here. It should be pointed out that for those skilled in the art, without departing from the technical solution of the present invention, several modifications and improvements can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. A method for preparing a cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide, characterized in that: The preparation method comprises the following steps: S1. Preparation of cuprous oxide: Prepare a first reaction container, add deionized water into the first reaction container, then add copper chloride dihydrate and polyvinyl pyrrolidone, stir for 1h to 2h, then add sodium hydroxide solution, stir again to dissolve the sodium hydroxide solution, introduce nitrogen into the first reaction container so that the first reaction container is in a nitrogen environment, then slowly add ascorbic acid, stir again for 1h to 2h, centrifuge the reactants, purify and dry the obtained solid to obtain cuprous oxide; S2. Preparation of cuprous selenide: Prepare a second reaction container, add deionized water into the second reaction container, introduce nitrogen into the second reaction container so that the second reaction container is in a nitrogen environment, add selenium powder and sodium borohydride, stir for 23h to 25h, then add cuprous oxide, stir for a second time, centrifuge the reactants after the reaction is completed, purify and dry the obtained solid to obtain cuprous selenide; S3, modified cuprous selenide: Prepare a third reaction container, add deionized water into the third reaction container, then add cuprous selenide and SH-PEG, stir, centrifuge the reactants after the reaction is completed, purify and dry the obtained solid to obtain modified cuprous selenide (SH-PEG-Cu2Se); S4. Preparation of calcium peroxide: Prepare a fourth reaction container, add hyaluronic acid into the fourth reaction container, stir the hyaluronic acid, add calcium chloride solution while stirring the hyaluronic acid, continue stirring for 23h to 25h, then add ammonia water and hydrogen peroxide in sequence, wait for the reaction for 2h to 4h, then place the reaction system in an ultrasonic environment, add sodium hydroxide to obtain a white emulsion containing HA-CaO2 nanoparticles, wash the corresponding HA-CaO2 nanoparticles with sodium hydroxide, water and ethanol in sequence, and then store the HA-CaO2 nanoparticles in ethanol at 2°C to 6°C; S5. Preparation of cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide: Prepare a fifth reaction container, add ethanol into the fifth reaction container, then add modified cuprous selenide and HA-CaO2, stir for 23h~25h, prepare a dissolution container, add ethanol into the dissolution container, then add lauric acid into the dissolution container, dissolve lauric acid in the ethanol in the dissolution container, then add the lauric acid ethanol mixture into the reaction container, stir for a second time for 4h~6h, after the reaction is completed, separate the reactants to obtain a precipitate, wash the precipitate with ethanol for 2~4 times, and obtain a cuprous selenide photothermochemical power nanocomposite loaded with calcium peroxide (Cu2Se-CaO2@LA).

2. The method for preparing a cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide according to claim 1, characterized in that: In step S1, the mass ratio of cupric chloride dihydrate, polyvinyl pyrrolidine and sodium hydroxide is 80-90:30:30-34.

3. The method for preparing a cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide according to claim 1, characterized in that: In the step S2, the mass ratio of selenium powder to sodium borohydride is 1-2:

1.

4. The method for preparing a cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide according to claim 1, characterized in that: In the step S3, the mass ratio of cuprous selenide to SH-PEG is 1 to 2:

1.

5. The method for preparing a cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide according to claim 1, characterized in that: In step S4, the concentration of the calcium chloride solution is 0.1 g / mL, the concentration of the hyaluronic acid solution is 15 mg / mL, and the volume ratio of the calcium chloride solution to the hyaluronic acid solution is 1:4-6.

6. The method for preparing a cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide according to claim 1, characterized in that: In step S5, the mass ratio of the modified cuprous selenide, HA-CaO2 and lauric acid is 1:0.8-1.2:98-102.

7. The method for preparing a cuprous selenide photothermochemical power nanoplatform loaded with calcium peroxide according to claim 1, characterized in that: In step S4, aqueous ammonia is added dropwise to adjust the pH value of the reaction system to 8.5-10.

8. Application of a cuprous selenide photothermochemical power nano-platform loaded with calcium peroxide prepared by the method according to any one of claims 1 to 6, characterized in that: Used for tumor treatment.

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