Copper-iron-sulfur-selenium nanoparticles, and preparation method and use thereof

Abstract

This invention relates to the pharmaceutical field, and particularly to copper-iron-sulfur-selenium nanoparticles, their preparation method, and applications. The preparation method includes the following steps: 1) mixing sulfur with a surfactant to alter the water solubility of sulfur; 2) mixing a selenium-containing compound with 1-octadecene to alter the water solubility of the selenium-containing compound; 3) mixing a copper-containing compound and an iron-containing compound with a surfactant to alter the water solubility of the copper-containing compound and the iron-containing compound; 4) reacting the mixture obtained in step 3) with the mixture obtained in steps 1) and 2) to obtain the copper-iron-sulfur-selenium nanoparticles. The copper-iron-sulfur-selenium nanoparticles of this invention can be used to treat cancers such as melanoma, integrating photothermal, pyroelectric, and chemodynamic therapy. This allows for the simultaneous synergistic effect of the three therapies, thereby enhancing anti-tumor efficacy, supplementing the limitations of single-therapy efficacy, improving tumor killing efficiency, and reducing toxic side effects, exhibiting high biocompatibility and safety.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and in particular to a copper-iron-sulfur-selenium nanoparticle, its preparation method, and its uses. Background Technology

[0002] Melanoma is a major disease threatening human life and health, with a high rate of disability and mortality, accounting for 79% of skin cancer deaths, imposing a huge economic burden on individuals, families, and society. Melanoma is a highly aggressive malignant skin cancer that can occur in the skin, choroid, and mucous membranes (respiratory, digestive, and genitourinary tracts). Melanoma is prone to spread or metastasis; if not removed early, it can rapidly metastasize to distant sites, resulting in extremely low survival rates. Currently, surgery combined with radiotherapy and chemotherapy is the main treatment for melanoma. However, this approach has significant limitations and substantial toxic side effects, such as surgical skin defects, systemic toxicity, tolerability issues, and damage to normal tissues. Therefore, there is an urgent need to develop a new, effective, and safe treatment method.

[0003] Photothermal therapy utilizes nanomaterials to convert absorbed near-infrared light energy into heat energy, effectively killing tumors. It is non-invasive, has high penetrability, and can be spatiotemporally controlled, thus holding great potential in cancer treatment. However, the efficacy of photothermal therapy alone is limited. Cancer cells can secrete heat shock proteins, developing heat tolerance, and excessively high temperatures can damage normal tissue structures. Therefore, developing a treatment that combines photothermal therapy with other therapies to enhance efficacy and reduce toxic side effects is of great significance. Summary of the Invention

[0004] In view of the shortcomings of the prior art described above, the purpose of this invention is to provide copper-iron-sulfur-selenium nanoparticles, their preparation method and uses, in order to solve the problems in the prior art.

[0005] To achieve the above and other related objectives, the present invention provides a method for preparing copper-iron-sulfur-selenium nanoparticles, the method comprising the following steps:

[0006] 1) Mix sulfur with surfactants to change the water solubility of sulfur;

[0007] 2) Mixing selenium-containing compounds with 1-octadecene to alter the water solubility of the selenium-containing compounds;

[0008] 3) Mixing copper-containing and iron-containing compounds with surfactants to alter the water solubility of the copper-containing and iron-containing compounds;

[0009] 4) The mixture obtained in step 3) is mixed with the mixture obtained in steps 1) and 2) and reacted to obtain copper-iron-sulfur-selenium nanoparticles.

[0010] Preferably, the surfactant in steps 1) and 3) is selected from cationic surfactants. Further, the cationic surfactant is selected from amines, more specifically from fatty amines, and even more specifically from oleylamines.

[0011] Preferably, the selenium-containing compound in step 2) is selected from SeO2.

[0012] Preferably, in step 3), the copper-containing compound is selected from CuCl2, and the iron-containing compound is selected from Fe(acac)3.

[0013] Preferably, the preparation method further includes hydrophilic modification of the copper-iron-sulfur-selenium nanoparticles.

[0014] The present invention also provides copper-iron-sulfur-selenium nanoparticles obtained by the preparation method described above.

[0015] The present invention also provides the use of the copper-iron-sulfur-selenium nanoparticles in the preparation of tumor treatment products.

[0016] The tumor is selected from one or more of the following: skin cancer, lung cancer, breast cancer, ovarian cancer, osteosarcoma, liver cancer, pancreatic cancer, prostate cancer, colorectal cancer, non-small cell lung cancer, kidney cancer, head and neck cancer, melanoma, and multiple myeloma.

[0017] The present invention also provides the use of the copper-iron-sulfur-selenium nanoparticles in the preparation of products that induce the generation of reactive oxygen species or hydroxyl radicals.

[0018] The present invention also provides a pharmaceutical composition comprising the copper-iron-sulfur-selenium nanoparticles and a pharmaceutically acceptable carrier or excipient.

[0019] As described above, the copper-iron-sulfur-selenium nanoparticles, their preparation method, and their uses of the present invention have the following beneficial effects:

[0020] 1. It can be administered by injection, which facilitates precise local treatment of tumors, improves anti-tumor efficacy, and has good biocompatibility, reducing toxic side effects.

[0021] 2. For the first time, copper-iron-sulfur-selenium nanosheets were used as pyroelectric catalysts in near-infrared light (NIR-1) for synergistic photothermal-pyroelectric-chemodynamic therapy of tumors. Copper-iron-sulfur-selenium nanosheets possess excellent photothermal conversion properties, generating thermal energy to ablate tumors under 808nm laser irradiation. Furthermore, the temperature difference generated by photothermal heating induces charge separation, forming an electric current and inducing the generation of reactive oxygen species (ROS), which can directly attack tumor cells. Thirdly, copper and iron ions can convert hydrogen peroxide into hydroxyl radicals through the Fenton reaction, thereby killing tumor cells. These three effects synergistically enhance the efficacy of tumor treatment. Modification with dimercaptosuccinic acid increases the water solubility of the copper-iron-sulfur-selenium nanosheets, giving them high hydrophilicity and excellent biocompatibility, allowing for convenient local injection into tumors.

[0022] 3. This invention can be used to treat cancers such as melanoma. Compared with existing treatment strategies, the advantages of this invention lie in its integration of photothermal, pyroelectric, and chemidynamic therapy into a single material. This allows for the simultaneous synergistic effect of the three therapies, thereby enhancing anti-tumor efficacy, overcoming the limitations of single-therapy efficacy, improving tumor-killing efficiency, reducing toxic side effects, and exhibiting high biocompatibility. The synergistic therapy does not solely rely on thermal ablation of tumor cells; it combines thermal ablation with ROS attack on tumor cells, thus reducing the temperature of pure photothermal therapy, minimizing damage to normal tissues caused by high temperatures, and increasing the safety of tumor treatment. Attached Figure Description

[0023] Figure 1 The images shown are scanning electron microscope (SEM) images (a) and transmission electron microscope (TEM) images (b) of copper-iron-sulfur-selenium nanosheets.

[0024] Figure 2 The elemental analysis energy spectrum of the copper-iron-sulfur-selenium nanosheets is shown.

[0025] Figure 3 The photothermal effect and stability of copper-iron-sulfur-selenium nanosheets are shown. (a) is a graph showing the change of solution temperature of copper-iron-sulfur-selenium nanosheets with different concentrations under NIR I region (808nm) laser radiation of 1.0W / cm2 with irradiation time; (b) is a graph showing the change of solution temperature of copper-iron-sulfur-selenium nanosheets with different light intensities with irradiation time; (c) to detect the photothermal stability of copper-iron-sulfur-selenium nanosheets, the material was repeatedly irradiated and then removed from the light source 5 times, and the temperature change value was recorded.

[0026] Figure 4 The image shows the UV characteristic absorption peaks of MB after heat treatment of copper-iron-sulfur-selenium nanosheets with added MB; "Materials" in the figure represents copper-iron-sulfur-selenium nanosheets.

[0027] Figure 5The image shows the UV absorption peaks of copper-iron-sulfur-selenium nanosheets with added DPBF after heat treatment; "Materials" in the figure represents copper-iron-sulfur-selenium nanosheets.

[0028] Figure 6 The image shows the electron spin resonance (ESR) pattern of copper-iron-sulfur-selenium nanosheets mixed with DMPO spin inducer under heating.

[0029] Figure 7 The image shows the electron spin resonance (ESR) pattern at room temperature after copper-iron-sulfur-selenium nanosheets are mixed with hydrogen peroxide and DMPO spin inducer.

[0030] Figure 8 The results show the cell activity of normal cells and tumor cells after 12 hours of incubation with copper, iron, sulfur and selenium nanosheets of different concentrations.

[0031] Figure 9 The figure shows the effect of different treatments on tumor cell activity as detected by CCK8; "Control", "NP" and "nanoparticle + laser group" in the figure represent blank control, copper-iron-sulfur-selenium nanosheet group and copper-iron-sulfur-selenium nanosheet + near-infrared I region 808nm laser irradiation group, respectively.

[0032] Figure 10 The image shows the survival of tumor cells after different treatments using live and dead dyes: green marks live cells and red marks dead cells; "Control", "NP", "NIR", and "NP+NIR" in the image represent blank control, copper-iron-sulfur-selenium nanosheet group, near-infrared I region 808nm laser irradiation group, and copper-iron-sulfur-selenium nanosheet + near-infrared I region 808nm laser irradiation group, respectively.

[0033] Figure 11 The figure shows the ROS level of tumor cells after different treatments detected by the DCFH probe. In the figure, "control group", "nanoparticle group", "laser group" and "nanoparticle + laser group" represent blank control, copper iron sulfur selenium nanosheet group, near-infrared I region 808nm laser irradiation group and copper iron sulfur selenium nanosheet + near-infrared I region 808nm laser irradiation group, respectively.

[0034] Figure 12 The in vivo antitumor properties of copper-iron-sulfur-selenium nanosheets are shown in (a) as the tumor volume changes over time after different treatments, and (b) as the tumor weight after different treatments. In the figure, “Control”, “NP”, “NIR”, and “NP+NIR” represent blank control, copper-iron-sulfur-selenium nanosheet group, near-infrared I region 808nm laser irradiation group, and copper-iron-sulfur-selenium nanosheet + near-infrared I region 808nm laser irradiation group, respectively. Detailed Implementation

[0035] This application provides a method for preparing copper-iron-sulfur-selenium nanoparticles (or copper-iron-sulfur-selenium nanosheets), the preparation method comprising the following steps:

[0036] 1) Mix sulfur with surfactants to change the water solubility of sulfur;

[0037] 2) Mixing selenium-containing compounds with 1-octadecene to alter the water solubility of the selenium-containing compounds;

[0038] 3) Mixing copper-containing and iron-containing compounds with surfactants to alter the water solubility of the copper-containing and iron-containing compounds;

[0039] 4) The mixture obtained in step 3) is mixed with the mixture obtained in steps 1) and 2) and reacted to obtain copper-iron-sulfur-selenium nanoparticles.

[0040] In some embodiments of the present invention, the surfactant in steps 1) and 3) is selected from cationic surfactants. Further, the cationic surfactant is selected from amines, more specifically from fatty amines, and even more specifically from oleylamines.

[0041] In some embodiments of the present invention, step 1) or step 3) is carried out in a solvent. Those skilled in the art can select a suitable type and amount of solvent to ensure that the reactants are sufficiently dispersed in the reaction system. The solvent can be an organic solvent. The solvent can be a solvent already present in commercially available surfactants for dissolving surfactants; it can be directly mixed with sulfur, copper-containing compounds, or iron-containing compounds during the reaction without the need for additional solvents.

[0042] In some embodiments of the present invention, the molar ratio of sulfur to surfactant is 1:5 to 1:15; preferably, it is 1:8 to 1:12; more preferably, it is 1:10.

[0043] In some embodiments of the present invention, step 1) further includes one or more of the following features:

[0044] 1a) The reaction is carried out under vacuum conditions;

[0045] 1b) The reaction temperature is 300–400 K; preferably, the reaction temperature is 330–370 K;

[0046] 1c) The reaction time is 5 to 7 hours; preferably, the reaction time is 5.5 to 6.5 hours.

[0047] In some embodiments of the present invention, the selenium-containing compound in step 2) is selected from SeO2.

[0048] In some embodiments of the present invention, the molar ratio of the selenium-containing compound and 1-octadecene in step 2) is 1:3 to 13; preferably, it is 1:6 to 1:10; more preferably, it is 1:8.

[0049] In some embodiments of the present invention, step 2) further includes one or more of the following features:

[0050] 2a) The reaction is carried out under vacuum conditions;

[0051] 2b) The reaction temperature is 300–400 K; preferably, the reaction temperature is 330–370 K;

[0052] 2c) The reaction time is 5 to 7 hours; preferably, the reaction time is 5.5 to 6.5 hours.

[0053] In step 3), the copper-containing compound is selected from CuCl2. The iron-containing compound is selected from Fe(acac)3 or Fe(NO3)3.

[0054] In some embodiments of the present invention, the molar ratio of the copper-containing compound, the iron-containing compound and the surfactant in step 3) is 5:1:(40-50), preferably 5:1:46.

[0055] In some embodiments of the present invention, step 3) further includes one or more of the following features:

[0056] 3a) The reaction is carried out under an inert gas atmosphere;

[0057] 3b) The reaction temperature is 250–600 K; preferably, the reaction is carried out at 250–350 K for 5–7 hours, and then the temperature is raised to 400–500 K for 5–15 minutes.

[0058] In some embodiments of the present invention, in step 4), the mixture obtained in step 3) is heated to 500-600K and then immediately mixed with the mixture obtained in steps 1) and 2).

[0059] In some embodiments of the present invention, the reaction temperature in step 4) is 500-600K; and / or the reaction time is 20-40 minutes.

[0060] In some embodiments of the present invention, step 4) further includes washing the copper-iron-sulfur-selenium nanoparticle product after cooling following the completion of the reaction. Specifically, the product is washed sequentially with hexane and ethanol.

[0061] In some embodiments of the present invention, the preparation method further includes hydrophilic modification of copper-iron-sulfur-selenium nanoparticles.

[0062] The hydrophilic modification involves mixing a hydrophilic reagent with copper, iron, sulfur, and selenium nanoparticles to conduct a ligand exchange reaction. The hydrophilic reagent is selected from DMSA.

[0063] In some embodiments of the present invention, the molar ratio of the hydrophilic reagent to the copper-iron-sulfur-selenium nanoparticles is 1:5 to 1:15; preferably, it is 1:10.

[0064] In some embodiments of the present invention, the hydrophilic modification step comprises: mixing a hydrophilic reagent with copper-iron-sulfur-selenium nanoparticles for 10–15 hours, preferably 11–13 hours, followed by ultrasonic treatment for 5–7 hours, preferably 5.5–6.5 hours, and then separating and washing to obtain water-soluble copper-iron-sulfur-selenium nanoparticles. In one embodiment, the ultrasonic frequency is 30–50 kHz; preferably 35–45 kHz; more preferably 40 kHz. In one embodiment, the nanoparticles are separated by centrifugation after ultrasonication. In one embodiment, the nanoparticles are repeatedly washed with water. In one embodiment, the obtained water-soluble copper-iron-sulfur-selenium nanoparticles can be dispersed in water for later use.

[0065] In some embodiments of the present invention, the hydrophilic modification is carried out in a solvent. Those skilled in the art can select suitable solvent types and amounts to ensure adequate dispersion of the reactants in the reaction system. The solvent may be an organic solvent. More specifically, the solvent may be selected from one or more of DMSO, hexane, DMF, DMAC, toluene, NMP, or pyridine. In one embodiment, the solvent is selected from DMSO and hexane in a volume ratio of 1:1.

[0066] The present invention also provides copper-iron-sulfur-selenium nanoparticles obtained by the preparation method described above.

[0067] The copper-iron-sulfur-selenium nanoparticles are coated with hydrophilic reagents such as dimercaptosuccinic acid to obtain water-soluble nanoparticles, enabling injection administration for convenient and precise local tumor treatment, improving anti-tumor efficacy, and exhibiting high biocompatibility and reduced toxic side effects. The copper-iron-sulfur-selenium nanoparticles also possess photothermal conversion properties.

[0068] The present invention also provides the use of the copper-iron-sulfur-selenium nanoparticles in the preparation of tumor treatment products.

[0069] The tumor is selected from one or more of the following: skin cancer, lung cancer, breast cancer, ovarian cancer, osteosarcoma, liver cancer, pancreatic cancer, prostate cancer, colorectal cancer, non-small cell lung cancer, kidney cancer, head and neck cancer, melanoma, and multiple myeloma.

[0070] The copper-iron-sulfur-selenium nanoparticles ablate tumors by generating heat energy through photothermal conversion. Secondly, they attack tumor cells by generating reactive oxygen species induced by the temperature difference caused by photothermal heating. Thirdly, the copper and iron ions in the copper-iron-sulfur-selenium nanoparticles themselves convert hydrogen peroxide into hydroxyl radicals that kill tumor cells.

[0071] The tumor treatment product may be a pharmaceutical composition.

[0072] The present invention also provides the use of the copper-iron-sulfur-selenium nanoparticles in the preparation of products that induce the generation of reactive oxygen species or hydroxyl radicals.

[0073] The product that induces the generation of reactive oxygen species or hydroxyl radicals may be a pharmaceutical composition.

[0074] The present invention also provides a pharmaceutical composition comprising the copper-iron-sulfur-selenium nanoparticles and a pharmaceutically acceptable carrier or excipient.

[0075] "Pharmaceutical acceptable" means that when a drug is properly administered to animals or humans, it will not produce adverse, allergic, or other adverse reactions.

[0076] A "pharmaceutically acceptable carrier or excipient" should be compatible with the active ingredient, meaning it can be miscible with it without significantly reducing the drug's efficacy under normal circumstances. Specific examples of substances that can serve as pharmaceutically acceptable carriers or excipients include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium methylcellulose, ethylcellulose, and methylcellulose; tragacanth gum powder; malt; gelatin; talc; solid lubricants such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter; polyols such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers such as Tween; wetting agents such as sodium lauryl sulfate; colorants; flavoring agents; tableting agents; stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic salt solutions; and phosphate buffers, etc. These substances are used as needed to help stabilize the formulation or to improve its activity or bioavailability or to produce an acceptable taste or smell when taken orally.

[0077] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0078] Before further describing specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terminology used in the embodiments of the present invention is for describing specific embodiments and not for limiting the scope of protection of the present invention; in the specification and claims of the present invention, unless otherwise expressly stated in the text, the singular forms "a", "an" and "this" include the plural forms.

[0079] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the present invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art. In addition to the specific methods, apparatus, and materials used in the embodiments, based on the knowledge of the prior art possessed by one of ordinary skill in the art and the description of this invention, any prior art methods, apparatus, and materials similar to or equivalent to those described, apparatus, and materials in the embodiments of this invention may be used to implement the present invention.

[0080] Example 1: Synthesis and Modification of Cu5FeS 3.6 Se 0.4 Nanoparticles

[0081] Synthesis of Cu5FeS 3.6 Se 0.4 Nanoparticles: 45 mmol / L S and 150 mL of oleylamine (OLA, 0.46 mol / L) were added to a 500 mL round-bottom flask. The solution was evacuated for 10 minutes, heated to 353 K, and then maintained at 353 K for 6 hours. Simultaneously, 4 mmol / L SeO2 and 10 mL of 1-octadecene (ODE, 30 mmol / L) were added to a 50 mL round-bottom flask. The solution was evacuated for 10 minutes, heated to 353 K, and maintained at 353 K for 6 hours. Simultaneously, 50 mmol CuCl2 and 10 mmol Fe(acac)3 were added to 100 mL OLA in a 1 L three-necked round-bottom flask. A nitrogen atmosphere was maintained throughout the process in the three-necked round-bottom flask to remove other gases. The solution was stirred at 306 K for 6 hours, heated to 453 K, and held for 10 minutes. After heating the CuCl2-Fe(acac)3-OLA solution to 553 K, the prepared S-OLA and SeO2-ODE solutions were rapidly poured into the OLA solution of the CuCl2-Fe(acac)3-OLA metal salt. The mixed solution was held at 553 K for 30 minutes and then naturally cooled to 333 K. The product was washed five times sequentially with hexane and ethanol, and finally dispersed in 25 mL of hexane.

[0082] Modified Cu5FeS 3.6 Se0.4 Nanoparticles: To improve Cu5FeS 3.6 Se 0.4 The compatibility of nanoparticles in biomedical applications, through hydrophilic DMSA and hydrophobic Cu5FeS 3.6 Se 0.4 Nanoparticles further modify Cu5FeS through ligand exchange reactions. 3.6 Se 0.4 Nanoparticles. 250 mg of DMSA was dissolved in 25 ml of DMSO (dimethyl sulfoxide). This solution was then mixed with a Cu5FeS3.6Se0.4 nanoparticle solution dissolved in hexane. The mixture was magnetically stirred for 12 h, followed by ultrasonication for 6 h. The product was separated by centrifugation and repeatedly washed with deionized water to obtain water-soluble DMSA-coated Cu5FeS3.6Se0.4 nanoparticles. 3.6 Se 0.4 @DMSA nanoparticles were redispersed in water. Cu5FeS 3.6 Se 0.4 @DMSA nanoparticles were examined using scanning electron microscopy and transmission electron microscopy, respectively, and the results are as follows: Figure 1 As shown, icosahedral nanoparticles are visible. Simultaneously, high-angle annular dark-field (HAADF) images and corresponding energy-dispersive X-ray spectra (EDS) were obtained, as shown... Figure 2 As shown, the icosahedral structure in the sample was confirmed, as well as the uniform Se distribution and iron-rich cores in the icosahedral particles.

[0083] Example 2. Detection of photothermal effect and stability

[0084] Cu5FeS 3.6 Se 0.4 @DMSA (800, 400, 200, 100 and 0 μg / mL) nanoparticles were dispersed in 96-well plates and subjected to a power density of 1.0 W / cm². 2 The water was irradiated with a near-infrared laser in the NIR I region (808 nm) for ten minutes, and the water temperature change over time was recorded using an infrared camera. The power density-dependent temperature curves were then obtained by applying different power densities (0.5, 0.75, 1, and 1.25 W / cm²). 2 808nm laser irradiation of Cu5FeS 3.6 Se 0.4 @DMSA (800 μg / mL) was obtained over 10 minutes. To check the photothermal conversion efficiency, Cu5FeS... 3.6 Se 0.4 @DMSA (800 μg / mL) was irradiated with an 808 nm laser for 5 minutes and then cooled. To test the photothermal stability of the material, the irradiation was repeated 5 times, and the temperature change of the water after each irradiation was recorded. The results are as follows: Figure 3 As shown, Cu5FeS 3.6 Se 0.4 @DMSA has good photothermal performance and photothermal stability.

[0085] Example 3. Testing the performance of pyroelectric and chemodynamic therapy

[0086] Cu5FeS 3.6 Se 0.4 @DMSA was dispersed in a centrifuge tube to obtain Cu5FeS 3.6 Se 0.4 The final concentration of DMSA was 800 μg / mL, then MB was added at a concentration of 10 μg / mL, for a total volume of 1 mL. The mixture was then heated to 65 °C, while a comparative experiment was conducted using a pure MB solution under the same conditions. The results are as follows... Figure 4 As shown, this confirms Cu5FeS 3.6 Se 0.4 @DMSA can generate hydroxyl radicals when heated.

[0087] Cu5FeS 3.6 Se 0.4 @DMSA was dispersed in a centrifuge tube to obtain Cu5FeS 3.6 Se 0.4 The final concentration of DMSA was 800 μg / mL, then DPBF was added to a concentration of 10 μg / mL, for a total volume of 1 mL. The mixture was then heated to 65 °C, while a comparative experiment was conducted using a pure DPBF solution under the same conditions. The results are as follows... Figure 5 As shown, this confirms Cu5FeS 3.6 Se 0.4 @DMSA can produce singlet oxygen when heated.

[0088] Further verification of Cu5FeS using electron spin resonance (ESR) technology 3.6 Se 0.4 @DMSA generates reactive oxygen species (superoxide radicals) upon heating. 1 mL of Cu5FeS... 3.6 Se 0.4 @DMSA was mixed with 1 μL of 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and heated at 65°C for 5 minutes. The efficacy of chemodynamic therapy was assessed by ESR, and the results are as follows. Figure 6 As shown, this confirms Cu5FeS 3.6 Se 0.4 @DMSA can generate superoxide radicals upon heating. Cu5FeS 3.6 Se0.4 @DMSA was mixed with 50 μL of hydrogen peroxide (final concentration of hydrogen peroxide was 1 mM), then mixed with 4 μL of DMPO, and incubated at room temperature for ten minutes before detection. The results are as follows. Figure 7 As shown, this confirms Cu5FeS 3.6 Se 0.4 @DMSA can use H2O2 as a reactant to trigger Fenton or Fenton-like reactions in tumor cells, catalyzing the production of hydroxyl radicals from H2O2, thereby mediating chemokinetic therapy.

[0089] Example 4. Detection of biocompatibility of nanoparticles

[0090] The cytotoxicity of different concentrations of copper-iron-sulfur-selenium nanosheets (0, 10, 20, 40, 80, and 160 μg / mL) to cells was evaluated using the CCK-8 assay. B16F10 and NIH / 3T3 cells were seeded in 6-well plates (2 x 10⁶ cells per well). 5 Cells were cultured overnight in a microplate solution containing copper, iron, sulfur, and selenium nanosheets, and then incubated for another 8 hours. CCK-8 solution was then added, and cell viability was assessed 4 hours later using a microplate reader at 450 nm absorbance. Figure 8 It can be seen that different concentrations of copper-iron-sulfur-selenium nanosheets had no significant effect on cell viability after 8 hours of incubation in B16F10 and NIH / 3T3 cells, indicating that the toxicity of copper-iron-sulfur-selenium nanosheets to cells is negligible, and that copper-iron-sulfur-selenium nanosheets exhibit excellent biocompatibility within the experimental concentration range.

[0091] Example 5. Detection of tumor cell killing effect by CCK-8 assay

[0092] B16F10 tumor cells were divided into three groups according to different treatment methods (copper-iron-sulfur-selenium nanosheet group only, copper-iron-sulfur-selenium nanosheet group + near-infrared I region 808nm laser irradiation group). Tumor cells were seeded into 96-well plates (2*10 wells). 4 Cells were cultured overnight in a solution of copper, iron, sulfur, and selenium nanosheets, and then incubated for 2 hours. Afterward, different treatments were performed using the same method. Finally, after co-incubation with CCK-8 solution for 4 hours, cell viability was assessed using a microplate reader with an absorbance of 450 nm. Figure 9 The results showed that copper-iron-sulfur-selenium nanosheets can kill tumor cells when irradiated with an 808nm laser in the near-infrared I region.

[0093] Example 6. Detection of the killing effect of live and dead dyes on tumor cells

[0094] B16F10 tumor cells were divided into four groups (blank control, copper-iron-sulfur-selenium nanosheet group, near-infrared I region 808nm laser irradiation group, and copper-iron-sulfur-selenium nanosheet + near-infrared I region 808nm laser irradiation group). Tumor cells were seeded into 6-well plates at 2*105 cells per well and cultured overnight, and then subjected to different treatments as described above. Figure 10 The results showed that a large number of dead cells could be observed in the group of copper-iron-sulfur-selenium nanosheets irradiated with 808nm laser in the near-infrared I region, proving that copper-iron-sulfur-selenium nanosheets can significantly kill tumor cells under laser irradiation.

[0095] Example 7. DCFH probe detection of ROS levels after different treatments

[0096] B16F10 tumor cells were divided into four groups (blank control, copper-iron-sulfur-selenium nanosheet group, near-infrared I region 808nm laser irradiation group, and copper-iron-sulfur-selenium nanosheet + near-infrared I region 808nm laser irradiation group). Tumor cells were seeded into 6-well plates at a density of 2*10 cells per well. 5 Cells were cultured overnight and then treated in different ways as described above. Figure 11 The results showed that copper-iron-sulfur-selenium nanosheets combined with 808nm near-infrared I region laser irradiation produced strong ROS, demonstrating that copper-iron-sulfur-selenium nanosheets significantly increased the ROS level in tumor cells under laser irradiation.

[0097] Example 8. In vivo anti-tumor assessment

[0098] B16F10 cells (1×10) 6 A melanoma model was established by subcutaneously injecting 100 μL of PBS into Balb / c mice. The tumors were then cultured until they reached approximately 100-150 mm in size. 3 Treatment began at a certain time, and 20 mice were randomly divided into different groups (n=5) for different treatments. The groups were: blank control (PBS), copper-iron-sulfur-selenium nanosheet group, near-infrared I region 808nm laser irradiation group, and copper-iron-sulfur-selenium nanosheet + near-infrared I region 808nm laser irradiation group. Tumor size was measured in each mouse every two days after treatment, and on day 12, tumors in each treatment group were dissected and weighed. Experimental results are as follows: Figure 12 As shown, the results indicate that the growth of tumor volume in tumor-bearing mice in the copper-iron-sulfur-selenium nanosheet + laser irradiation group was significantly inhibited.

[0099] The above embodiments are for illustrating the implementation schemes disclosed in this invention and should not be construed as limiting the invention. Furthermore, various modifications and variations of the methods listed herein will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been specifically described in conjunction with various specific preferred embodiments, it should be understood that the invention should not be limited to these specific embodiments. In fact, various modifications as described above that are obvious to those skilled in the art to obtain the invention should be included within the scope of this invention.

Claims

1. A method for preparing copper-iron-sulfur-selenium nanoparticles, characterized in that, The preparation method includes the following steps: 1) Mix sulfur with surfactants to alter its water solubility; 2) Mixing a selenium-containing compound with 1-octadecene to alter the water solubility of the selenium-containing compound; the selenium-containing compound is selected from SeO2; 3) Mix copper-containing compounds and iron-containing compounds with surfactants to change the water solubility of copper-containing compounds and iron-containing compounds; the molar ratio of copper-containing compounds, iron-containing compounds and surfactants is 5:1:(40~50); 4) The mixture obtained in step 3) is mixed with the mixture obtained in steps 1) and 2) and reacted to obtain copper-iron-sulfur-selenium nanoparticles.

2. The preparation method according to claim 1, characterized in that, The surfactants used in steps 1) and 3) are selected from cationic surfactants.

3. The preparation method according to claim 2, characterized in that, The cationic surfactant is selected from amines.

4. The preparation method according to claim 3, characterized in that, The amines mentioned are fatty amines.

5. The preparation method according to claim 4, characterized in that, The fatty amine is oleylamine.

6. The preparation method according to claim 1, characterized in that, Step 1) also includes one or more of the following features: 1a) The molar ratio of sulfur to surfactant is 1:5 to 1:15; 1b) The reaction temperature is 300~400K; 1c) The reaction time is 5-7 hours; 1d) React under vacuum conditions.

7. The preparation method according to claim 6, characterized in that, The molar ratio of sulfur to surfactant is 1:8 to 1:12; And / or, the reaction temperature is 330~370K; And / or, the reaction time is 5.5 to 6.5 hours.

8. The preparation method according to claim 1, characterized in that, Step 2) also includes one or more of the following features: 2a) The reaction is carried out under vacuum conditions; 2b) The reaction temperature is 300~400K; 2c) The reaction time is 5-7 hours; 2d) The molar ratio of selenium-containing compounds to 1-octadecene is 1:3~13.

9. The preparation method according to claim 8, characterized in that, The reaction temperature is 330~370K; And / or, the reaction time is 5.5 to 6.5 hours; And / or, the molar ratio of selenium-containing compound to 1-octadecene is 1:6 to 1:

10.

10. The preparation method according to claim 1, characterized in that, In step 3), the copper-containing compound is selected from CuCl2, and / or the iron-containing compound is selected from Fe(acac)3.

11. The preparation method according to claim 1, characterized in that, Step 3) also includes one or more of the following features: 3a) The reaction is carried out under an inert gas atmosphere; 3b) The reaction temperature is 250~600K.

12. The preparation method according to claim 11, characterized in that, The reaction temperature is 250~350K for 5~7 hours, and then the temperature is raised to 400~500K for 5~15 minutes.

13. The preparation method according to claim 1, characterized in that, In step 4), the mixture obtained in step 3) is heated to 500-600K and then immediately mixed with the mixture obtained in steps 1) and 2), and / or the reaction time after mixing is 20-40 minutes.

14. The preparation method according to claim 1, characterized in that, Step 4) also includes washing the copper, iron, sulfur, and selenium nanoparticle product after cooling following the completion of the reaction.

15. The preparation method according to claim 14, characterized in that, Wash with hexane and ethanol in sequence.

16. The preparation method according to claim 1, characterized in that, The preparation method also includes hydrophilic modification of copper-iron-sulfur-selenium nanoparticles.

17. The preparation method according to claim 16, characterized in that, The hydrophilic modification involves mixing a hydrophilic reagent with copper, iron, sulfur, and selenium nanoparticles to carry out a ligand exchange reaction.

18. The preparation method according to claim 17, characterized in that, The molar ratio of the hydrophilic reagent to the copper-iron-sulfur-selenium nanoparticles is 1:5 to 1:

15.

19. The preparation method according to claim 18, characterized in that, The hydrophilic reagent is selected from DMSA.

20. The preparation method according to claim 16, characterized in that, The hydrophilic modification steps are as follows: the hydrophilic reagent is stirred and mixed with copper-iron-sulfur-selenium nanoparticles for 10-15 hours, followed by ultrasonic treatment for 5-7 hours, and then separated to obtain water-soluble copper-iron-sulfur-selenium nanoparticles.

21. Copper-iron-sulfur-selenium nanoparticles obtained by the preparation method according to claims 1-20.

22. Use of the copper-iron-sulfur-selenium nanoparticles of claim 21 in the preparation of tumor treatment products.

23. The use according to claim 22, characterized in that, The tumor is selected from one or more of the following: skin cancer, lung cancer, breast cancer, ovarian cancer, osteosarcoma, liver cancer, pancreatic cancer, prostate cancer, colorectal cancer, non-small cell lung cancer, kidney cancer, head and neck cancer, melanoma, and multiple myeloma.

24. Use of the copper-iron-sulfur-selenium nanoparticles of claim 21 in the preparation of products that induce the generation of reactive oxygen species or hydroxyl radicals.

25. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the copper-iron-sulfur-selenium nanoparticles of claim 21 and a pharmaceutically acceptable carrier or excipient.

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