Tellurium-based nano material capable of efficiently inducing copper death as well as preparation method and application of tellurium-based nano material

Abstract

The invention discloses a tellurium-based nano material capable of efficiently inducing copper death as well as a preparation method and application of the tellurium-based nano material. The preparation method comprises the following steps: by taking water as a medium, mixing soluble copper salt and a sulfydryl-containing compound, and stirring to react, so as to obtain a Cu-S2 solution; dropwise adding the Cu-S2 solution into the TeO2 nano-carrier solution, and carrying out stirring reaction; and after the reaction is finished, carrying out solid-liquid separation, taking the solid, and resuspending with methanol to obtain the TeO2 coated Cu-S2 nano material. The preparation method is simple and rapid, large-scale production can be achieved, and the obtained TeO2 coated Cu-S2 nano material can efficiently induce cancer cells to generate copper death, is effective to various cancer cells and has potential clinical transformation application prospects.

Description

Technical Field

[0001] This invention belongs to the fields of nanomaterial chemistry and biomedicine, and specifically relates to a tellurium-based nanomaterial that efficiently induces copper death, its preparation method, and its application. Background Technology

[0002] Radiotherapy is a crucial treatment for malignant tumors, its mechanism primarily relying on the precise irradiation of diseased tissues by ionizing radiation. On one hand, radiation can directly cause DNA damage in tumor cells; on the other hand, it can indirectly lead to DNA breaks and dysfunction by generating a large number of reactive free radicals through the ionization of water molecules, thereby inhibiting tumor cell proliferation and inducing cell death. However, current radiotherapy still faces significant limitations: firstly, some tumor cells have low radiation sensitivity and are prone to developing radiation tolerance, leading to decreased treatment efficacy; secondly, despite continuous advancements in radiotherapy technology, achieving completely precise tumor-targeted irradiation remains difficult, while protecting surrounding normal tissues remains a challenge. Therefore, developing novel, highly efficient, and low-toxicity radiosensitizers to enhance tumor cell radiosensitivity and reduce toxic side effects has become a critical scientific problem urgently needing to be solved in the field of radiotherapy, possessing significant clinical translational value and broad application prospects.

[0003] Recent studies have shown that copper death (Cuproptosis) is a newly discovered form of copper-dependent programmed cell death, whose molecular mechanism mainly involves a cascade reaction triggered by an imbalance in intracellular copper ion homeostasis. Specifically, excess copper ions can specifically bind to esterified mitochondrial proteins in the tricarboxylic acid cycle (TCA), leading to abnormal oligomerization of esterified proteins and downregulation of iron-sulfur cluster protein expression. Ultimately, this induces severe protein toxicity stress, triggering an irreversible cell death program. In recent years, studies have found that the expression levels of key copper death regulatory proteins FDX1 and LIAS are significantly upregulated in residual lesions after radiotherapy of primary tumors. Therefore, targeting copper death provides a potential therapeutic target for reversing the radioresistance of residual tumor cells after radiotherapy.

[0004] However, copper ion homeostasis within tumor cells is strictly regulated, and their endogenous copper ion concentration is typically low, insufficient to effectively induce copper death. Therefore, the development of a copper ion delivery system capable of simultaneously enhancing intracellular copper ion concentration in tumor cells and improving radiotherapy efficacy is urgently needed. Summary of the Invention

[0005] The primary objective of this invention is to overcome the shortcomings and deficiencies of the prior art and provide a method for preparing tellurium-based nanomaterials that efficiently induce copper death.

[0006] Another object of the present invention is to provide tellurium-based nanomaterials that are highly efficient at inducing copper death, obtained by the above preparation method.

[0007] Another object of the present invention is to provide the application of the above-mentioned tellurium-based nanomaterials that efficiently induce copper death.

[0008] The objective of this invention is achieved through the following technical solution: A method for preparing tellurium-based nanomaterials that efficiently induce copper death includes the following steps: (1) Using water as a medium, a soluble copper salt and a mercapto-containing compound are mixed and stirred to react. The resulting product is named Cu-S2 solution. (2) Cu-S2 solution was added dropwise to TeO2 nanocarrier solution and stirred to react; after the reaction was completed, solid and liquid were separated, the solid was taken and resuspended in methanol to obtain TeO2@Cu-S2 solution.

[0009] The soluble copper salt mentioned in step (1) is preferably copper chloride or copper chloride hydrate.

[0010] The thiol-containing compound mentioned in step (1) is preferably thiol succinic acid.

[0011] The amounts of the thiol-containing compound and the soluble copper salt in step (1) are preferably calculated based on a molar ratio of sulfur in the thiol-containing compound to copper in the soluble copper salt of not less than 2:1; more preferably based on a molar ratio of sulfur in the thiol-containing compound to copper in the soluble copper salt of 2.1 to 2.3:1.

[0012] The stirring speed in step (1) is preferably 200 to 400 rpm; more preferably 300 rpm.

[0013] The stirring reaction time in step (1) is preferably 5 to 15 min; more preferably 10 min.

[0014] The TeO2 nanocarrier solution described in step (2) is preferably prepared by the following steps: the pH of the soluble inorganic tellurium salt aqueous solution under stirring is adjusted to acidic, and the reaction is stirred; after the reaction is completed, the solid and liquid are separated, and the solution is resuspended in methanol to obtain the TeO2 nanocarrier solution.

[0015] The pH is adjusted by an acid; preferably by a weak acid.

[0016] The weak acid is preferably mercaptosuccinic acid.

[0017] The pH value is preferably 4.0 to 6.0; more preferably 4.5 to 5.5.

[0018] The soluble inorganic tellurium salt is preferably sodium tellurite.

[0019] The amount of acid used is preferably calculated as 9-11 mg of acid per 0.011 g sodium tellurite solution; more preferably, it is calculated as 10 mg of acid per 0.011 g sodium tellurite solution.

[0020] The concentration of the soluble inorganic tellurium salt aqueous solution is preferably 8 mmol / mL.

[0021] The reaction time is preferably 10 to 30 minutes; more preferably 20 minutes.

[0022] The preferred method for solid-liquid separation is centrifugation.

[0023] The preferred centrifugation conditions are 10,000–12,000 rpm for 5–15 minutes.

[0024] In step (2), the Cu-S2 and the TeO2 nanocarrier are preferably mixed in a copper-tellurium mass ratio of 75-85:1100-1200; more preferably in a copper-tellurium mass ratio of 80:1120.

[0025] The stirring speed in step (2) is preferably 200 to 400 rpm; more preferably 300 rpm.

[0026] The stirring reaction time in step (2) is preferably 0.5 to 2 h; more preferably 1 h.

[0027] The preferred method for solid-liquid separation in step (2) is centrifugation.

[0028] The preferred centrifugation conditions are 10,000–12,000 rpm for 5–15 minutes.

[0029] The water used in this invention is preferably ultrapure water.

[0030] Unless otherwise specified, the reaction is carried out at room temperature. The room temperature is 15–35 °C; preferably 20–30 °C; more preferably 24–26 °C.

[0031] A tellurium-based nanomaterial that efficiently induces copper death was prepared by the above method, with an average particle size of about 140 nm.

[0032] The aforementioned tellurium-based nanomaterials, which efficiently induce copper death, can simultaneously induce the aggregation of multiple cancer cell-related proteins, leading to copper death and thus inducing a powerful radioimmunotherapy effect. Simultaneously, these tellurium-based nanomaterials, by enhancing X-ray absorption, generate abundant reactive oxygen species (ROS), inducing significant DNA damage and thereby exerting a radiosensitizing effect.

[0033] The above-mentioned tellurium-based nanomaterials that efficiently induce copper death can be used in the preparation of X-ray radiosensitizers or anti-tumor drugs.

[0034] The tumor is preferably a cancer of the reproductive system.

[0035] The reproductive system cancer is preferably cervical cancer; more preferably cervical squamous cell carcinoma.

[0036] The present invention has the following advantages and effects compared with the prior art: 1. The tellurium-based nanomaterials for efficiently inducing copper death provided by this invention can efficiently induce copper death in cancer cells, are effective against a variety of cancer cells, and have versatility. Furthermore, the synthesis process is simple and rapid, enabling large-scale production, which makes these tellurium-based nanomaterials for efficiently inducing copper death even more promising for clinical translational applications.

[0037] 2. The tellurium-based nanomaterials provided by this invention can not only efficiently induce copper death, but also exhibit a more specific response than existing inducers in the tumor microenvironment.

[0038] 3. By utilizing the photoelectric effect and Compton scattering properties of tellurium, the tellurium-based nanomaterials that provide this invention can generate abundant ROS, induce a large amount of DNA damage, and thus exert a radiation sensitizing effect.

[0039] 4. The tellurium-based nanomaterials that provide efficient copper death induction provided by this invention work synergistically with other clinical treatments (radiotherapy, immunotherapy) to improve the efficacy of radioimmunotherapy, effectively inhibit the malignant progression of tumors, and provide a reference for the efficacy and mechanism of existing copper death combined with other therapies. Attached Figure Description

[0040] Figure 1 Transmission electron microscopy (TEM) images of TeO2@Cu and TeO2@Cu-S2.

[0041] Figure 2 TeO2@Cu-S 2、 Cytotoxicity test results for TeO2@Cu and MSA.

[0042] Figure 3 The image shows the test results of the killing ability of different materials against different tumor cells.

[0043] Figure 4 The figure shows the detection results of the ability of different materials to induce copper death in different tumor cells.

[0044] Figure 5 The image shows the test results of the radiosensitization ability of different materials.

[0045] Figure 6This is a graph showing the detection results of the in vivo immune activation capacity of different materials.

[0046] Figure 7 The results of generating reactive oxygen species (ROS) in cells for different materials are shown in the figure. Detailed Implementation

[0047] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0048] Example 1 (1) Weigh 0.0886 g of sodium tellurite and dissolve it in 50 mL of ultrapure water to obtain a sodium tellurite stock solution with a concentration of 8 mM. Weigh 100 mg of mercaptosuccinic acid (MSA) and dissolve it in 10 mL of ultrapure water to obtain an MSA stock solution with a concentration of 0.066 mmol / mL. Weigh 100 mg of CuCl2•2H2O and dissolve it in 10 mL of ultrapure water to obtain a copper chloride stock solution with a concentration of 0.03 mmol / mL. Weigh 50 mg of MSA and dissolve it in 10 mL of ultrapure water to obtain an MSA stock solution with a concentration of 0.033 mmol / mL. All prepared stock solutions were stored at 4 ℃ for later use.

[0049] (2) 1 mL of MSA stock solution with a concentration of 0.066 mmol / mL was slowly added dropwise to 6.25 mL of sodium tellurite stock solution with a concentration of 8 mM. Then, sufficient ultrapure water was added to bring the final volume to 25 mL. After reacting at room temperature for 20 min, the precipitate was collected by centrifugation at 12000 rpm for 10 min and resuspended in 4 mL of anhydrous methanol to obtain the TeO2 stock solution. According to the detection by liquid chromatography-inductively coupled plasma mass spectrometry (ICP), the concentration of Te was 280 µg / mL.

[0050] (3) 2 mL of 0.03 mmol / mL copper chloride stock solution was added dropwise to 8 mL of 0.033 mmol / mL MSA stock solution, and the reaction was carried out at 300 rpm for 10 min at room temperature. The product was named Cu-S2 solution. The concentration of Cu was 0.4 mg / mL according to the detection by liquid chromatography-inductively coupled plasma mass spectrometry (ICP).

[0051] (4) Add 200 µL of Cu-S2 solution prepared in step (3) dropwise to 4 mL of TeO2 solution prepared in step (2), react at 300 rpm at room temperature for 1 h, then centrifuge at 12000 rpm for 10 min to collect the precipitate, and then resuspend it in 4 mL of anhydrous methanol to obtain TeO2@Cu-S2 solution, wherein the concentration of Te is 280 µg / mL and the concentration of Cu is 22 µg / mL.

[0052] (5) Add 42 µL of copper chloride stock solution with a concentration of 0.03 mmol / mL dropwise to 4 mL of TeO2 solution prepared in step (2), react at a stirring speed of 300 rpm for 1 h, remove unreacted impurities by centrifugation at 12000 rpm for 10 min, and then resuspend in 4 mL of anhydrous methanol to obtain TeO2@Cu solution, wherein the concentration of Te is 280 µg / mL and the concentration of Cu is 22 µg / mL.

[0053] Example 1 The TeO2@Cu and TeO2@Cu-S2 prepared in Example 1 were observed by transmission electron microscopy. Figure 1 As shown, TeO2@Cu-S2 has a uniform particle size and good dispersion, with a particle size of about 140 nm; TeO2@Cu has a less uniform particle size and moderate dispersion, with a particle size of about 140 nm.

[0054] Example 2 (1) Human cervical adenocarcinoma cells HeLa, human cervical squamous cell carcinoma cells C33A, human cervical cancer intestinal metastasis cells Caski, and human cervical squamous cell carcinoma cells Siha (purchased from the American Center for Standard Biological Collections, ATCC) in logarithmic growth phase were respectively inoculated at 3×10 4 Cells were seeded at a density of 100 µL / well in 96-well plates and incubated overnight. Then, different concentrations of TeO2, TeO2@Cu, and TeO2@Cu-S2 (0.5, 1, 2, 4, and 8 µg / mL) were added to treat the cells (100 µL per well), and the plates were incubated for another 72 h. After 72 h, 30 µL of 5 mg / mL MTT solution was added to each well, and the plates were incubated for 3-4 hours. Then, 150 µL of DMSO was added, and the plates were shaken for 5 min to fully dissolve the blue-purple formazan crystals. The absorbance was then measured at 570 nm using a microplate reader. Cell viability was calculated using the formula: Cell viability = (Absorbance of drug-treated group) / (Absorbance of control group) × 100%. Finally, the half-maximal inhibitory concentration (IC50) of different drugs and different doses of radiotherapy on different cell types was calculated using Graphpad Prism 10.0 software. 50(Value) to assess the effect of drugs on cell growth.

[0055] The results are as follows Figure 2 As shown, TeO2@Cu-S2 exhibits concentration-dependent toxicity against various cervical cancer cell lines, with the most significant killing effect on Siha cells. Therefore, to enhance the specificity of the research, subsequent experiments can prioritize the use of Siha cells for in-depth investigation.

[0056] (2) The difference between TeO2@Cu and TeO2@Cu-S2 is that the former uses MSA stock solution with a concentration of 5 mg / mL and copper chloride solution to prepare Cu-S2 solution. Therefore, the specific experimental procedure for detecting the toxic effects of MSA on cells is as follows: Using 3×10 4 Siha cells were seeded at a density of 100 µL / mL into 96-well plates and incubated overnight. Cells were then treated with the corresponding MSA concentrations of TeO2@Cu and TeO2@Cu-S2 (100 µL / well) and incubated for another 72 h. After 72 h, 30 µL of 5 mg / mL MTT solution was added to each well, and the plates were incubated for 3–4 h. Then, 150 µL of DMSO was added, and the plates were shaken for 5 min to fully dissolve the blue-purple formazan crystals. The absorbance was then measured at 570 nm using a microplate reader. Cell viability was calculated using the formula: Cell viability = (Absorbance of the treated group) / (Absorbance of the control group) × 100%.

[0057] Figure 3 The results indicate that low concentrations of MSA have no significant toxicity to Siha cells, and the difference in toxicity between TeO2@Cu and TeO2@Cu-S2 is not significantly related to the difference in MSA concentration.

[0058] (3) Take Siha cells in the logarithmic growth phase and use 1×10 6Siha cells were seeded at a density of 10 cells / well in 10 cm culture dishes with 10 mL of culture medium and cultured overnight. After cell adhesion, TeO2@Cu and TeO2@Cu-S2 were added at a concentration of 2 µg / mL, respectively, and cultured for 48 h. Cell proteins were then collected. Cells were incubated with lysis buffer, and total cell protein was extracted. The protein concentration of Siha cells after treatment was detected by bovine serum albumin (BSA). An equal volume (20 µg) of protein was electrophoresed on a 10% SDS-PAGE polyacrylamide gel and then transferred to a polyvinylidene fluoride (PVDF) membrane, which was blocked with 5% skim milk in TBST buffer for 2 h. The membrane was then incubated overnight at 4 °C with the corresponding primary antibodies (rabbit anti-DLAT antibody, rabbit anti-FDX1 antibody, and mouse anti-GAPDH antibody) at a dilution of 1:1000. It was then incubated with secondary antibodies at 4 °C at a dilution of 1:3000 for 2 h and washed three times with TBST. Finally, the proteins were developed using a Kodak X-ray chemiluminescence analyzer. GAPDH was used to confirm the equal loading and transfer of proteins. FDX1 and DLAT are key proteins involved in copper death. The protein blot bands were analyzed using ImageJ software for grayscale analysis.

[0059] The results are as follows Figure 4 As shown, although X-ray irradiation alone induces upregulation of FDX1 protein, thus hindering copper death, the combination of X-rays with TeO2@Cu or TeO2@Cu-S2 significantly inhibits FDX1 expression and induces thioacrylation and oligomerization of the mitochondrial protein DLAT. Notably, the combination of X-rays and TeO2@Cu-S2 is significantly more effective than TeO2@Cu, indicating that it can more effectively activate the copper death pathway, thereby enhancing antitumor activity.

[0060] (4) Take Siha cells in the logarithmic growth phase at 3×10 4Cells were seeded at a density of 100 µL / mL into 96-well plates and incubated overnight. Then, different concentrations of TeO2@Cu-S2 (0.5, 1, 2, 4, 8 µg / mL) were added to each well (100 µL per well) for incubation for 4–6 hours. Afterward, the cells were irradiated at doses of 0, 2, and 4 Gy, and then incubated for another 72 hours. After this time, 30 µL of 5 mg / mL MTT solution was added to each well, and the cells were incubated for 3–4 hours. Then, 150 µL of DMSO was added, and the plate was shaken for 5 minutes to fully dissolve the blue-purple formazan crystals. The absorbance was then measured at 570 nm using a microplate reader. Cell viability was calculated using the formula: Cell viability = (Absorbance of treated group) / (Absorbance of control group) × 100%. Finally, Graphpad Prism 10.0 software was used to calculate the half-maximal inhibitory concentration (IC50) of different drugs and different radiotherapy doses on different cells. 50 (Value) to assess the effect of drugs on cell growth.

[0061] The results are as follows Figure 5 As shown: Figure 5 In this context, A represents the half-maximal inhibitory concentration (IC50) of the drug when TeO2, TeO2@Cu, TeO2@Cu-S2, and X-Ray are used in combination. With increasing TeO2@Cu-S2 concentration and X-Ray dose, cell proliferation is significantly inhibited and cell viability decreases in a dependent manner. Higher drug concentrations result in higher IC50 values ​​with higher radiotherapy doses. 50 The lower the value, the better the radiosensitization effect of TeO2@Cu-S2, indicating that TeO2@Cu-S2 and X-Ray have a significant synergistic killing effect on Siha cells; Figure 5 B in the graph represents the cell viability radar, which shows the cytotoxic effect of the combined treatment of TeO2@Cu-S2 and X-Ray on Siha cells. It demonstrates the dual dependence of cells on the concentration of TeO2@Cu-S2 and the dose of X-Ray irradiation, with the two synergistically enhancing cytotoxicity. Figure 5 In the figure, C represents the synergistic and antagonistic effects of TeO2@Cu-S2 combined with X-Ray. It can be seen that the drug TeO2@Cu-S2 produced a significant synergistic effect with radiotherapy at radiation doses of 2 Gy and 4 Gy, effectively enhancing the anti-tumor effect. This indicates that TeO2@Cu-S2 is a potential radiosensitizer.

[0062] Siha cells in the logarithmic growth phase were harvested at 3 × 10⁻⁶. 5Cells were seeded at a density of 100 cells / mL into laser confocal dishes, with 2 mL added to each well, and cultured for 24 h until adherence. TeO2@Cu-S2 was added to achieve a drug concentration of 1 µg / mL in the dish. After incubation for 5 h, cells requiring radiotherapy were treated with 4 Gy, and then incubated for another 8 h. The original culture medium was then aspirated, and the cells were washed three times with PBS. Mitochondrial dye Mito-Tracker Green and nuclear dye DAPI were then used for staining at a volume ratio of 1:1000. After incubation for 30 min, the dye was aspirated, and the cells were washed three times with PBS before imaging. Results are shown below. Figure 5 As shown in Figure D: Mitochondrial morphological analysis revealed that the mitochondrial structure of the control group cells was intact and clear, while treatment with single drug (TeO2@Cu-S2) or simple irradiation (X-Ray) induced significant mitochondrial fragmentation; the damage was most severe when both were used in combination. Given that mitochondria are the main site of copper death, this morphological result corroborates the results of FDX1 upregulation and DLAT oligomerization in Western blotting analysis, jointly confirming that the combination therapy successfully activated the copper death pathway by significantly disrupting mitochondrial structure, thereby exerting a potent antitumor effect.

[0063] (5) Resuspend Siha cells in PBS at a concentration of 1.5 × 10⁶ cells per mouse. 7 Cells per mL were injected subcutaneously into the hind legs of BALB / c-Nude nude mice (purchased from Guangdong Yaokang Biotechnology Co., Ltd.) at a density of 100 µL of cell solution per mouse to construct a subcutaneous xenograft model. The resulting tumors were then treated in groups as follows: Group G1 received no treatment for 21 days after tumor implantation; Group G2 received TeO2@Cu, Group G3 received TeO2@Cu-S2, Group G4 received radiotherapy, Group G5 received radiotherapy + TeO2@Cu, and Group G6 received radiotherapy + TeO2@Cu-S2. The drugs used were TeO2@Cu and TeO2@Cu-S2 solutions with a Te concentration of 280 µg / mL and a Cu concentration of 22 µg / mL, respectively. Each mouse was diluted with 0.04 µL of the aforementioned drug in 100 µL of PBS for each administration, with dilutions increased according to the number of mice. The tumor was administered via tail vein injection, with the drug given every other day for a total of 21 days, or 11 injections. The radiotherapy dose was 4 Gy per injection, once a week, for a total of three radiotherapy sessions. The tumor mass was then harvested and cryopreserved.

[0064] Remove the frozen tumor fragments and place them in a pre-chilled 50 mL centrifuge tube. Add 5-10 mL of pre-chilled PBS buffer and grind thoroughly on ice. Filter the resulting cell suspension through a 70 µm nylon filter into a new 2 mL centrifuge tube. Add 0.5 mL of 1× erythrocyte lysis buffer to the cell pellet, gently pipette to resuspend, and lyse on ice in the dark for 5 minutes. Then add sufficient PBS to stop lysis, centrifuge at 1500 rpm for 5 minutes, and discard the supernatant. Observe the cell pellet; if it is red, repeat the lysis step until a white cell pellet is obtained, indicating immune cells. Finally, resuspend the cells in 2.5 mL of PBS and perform cell counting. Based on the antibodies shown in the figure (the flow cytometry antibodies required for detecting NK1.1 are CD45 (BV421), NK1.1 (PE), and CD3 (FITC); the flow cytometry antibodies for detecting M1 immune cells are CD45 (BV421), CD11b (FITC), and CD86 (APC-CY7), the analysis of tumor-infiltrating immune cells was performed. According to the above protocol, the cell suspension was thoroughly mixed with the corresponding antibodies and incubated at 4°C in the dark for 30 minutes. After incubation, 1 mL of PBS was added to wash the cells, and the cells were centrifuged at 1500 rpm for 5 minutes, discarding the supernatant. Finally, the cells were resuspended in 300 µL of PBS, transferred through a filter to flow cytometry tubes, and immediately analyzed by flow cytometry for immune cells, including CD86, in tumors of different treatment groups. + T cells, CD4 + T is a detailed expression.

[0065] like Figure 6 As shown, TeO2@Cu-S2 combined with X-rays can effectively enhance the activation of NK cells, effectively induce the maturation of DC cells and promote the transformation of M2 into M1 macrophages, thereby activating the anti-tumor immune response.

[0066] (6) Siha cells (2×10 5 Siha cells were seeded in 96-well plates (cells / mL) and allowed to adhere overnight. Then, 1.0 µg / mL of the drug (100 µL) was added and the cells were incubated for 4 h. The irradiated group was then subjected to radiotherapy (4 Gy), followed by removal of the culture medium and washing with 200 µL of PBS. 10 µM of the DCFH-DA probe (100 µL) was incubated with the cells in the dark for 30 min. Subsequently, ROS levels in the cells were recorded using a multimodal fluorescent reader at excitation and emission wavelengths of 485 and 528 nm, respectively.

[0067] The results are as follows Figure 7As shown, treatment of Siha cells with STeO2, TeO2@Cu, and TeO2@Cu-S2 alone induced an increase in intracellular total reactive oxygen species (ROS) levels. When these treatments were combined with 4 Gy X-ray irradiation, a significant synergistic effect was observed, leading to a sharp increase in ROS levels. This result confirms that our developed drug can synergistically enhance radiosensitivity and strengthen antitumor efficacy by significantly increasing intracellular ROS levels in tumor cells.

[0068] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A method for preparing tellurium-based nanomaterials that efficiently induce copper death, characterized in that... It includes the following steps: (1) Using water as a medium, a soluble copper salt and a mercapto-containing compound are mixed and stirred to react. The resulting product is named Cu-S2 solution. (2) Cu-S2 solution was added dropwise to TeO2 nanocarrier solution and stirred to react; after the reaction was completed, solid and liquid were separated, the solid was taken and resuspended in methanol to obtain TeO2@Cu-S2 solution.

2. The method for preparing tellurium-based nanomaterials with high efficiency inducing copper death according to claim 1, characterized in that: The soluble copper salt mentioned in step (1) is copper chloride or copper chloride hydrate; The thiol-containing compound mentioned in step (1) is mercaptosuccinic acid; The TeO2 nanocarrier solution described in step (2) is prepared by the following steps: the pH of the soluble inorganic tellurium salt aqueous solution under stirring is adjusted to acidic, and the reaction is stirred; after the reaction is completed, the solid and liquid are separated, and the solution is resuspended in methanol to obtain the TeO2 nanocarrier solution.

3. The method for preparing tellurium-based nanomaterials with high efficiency inducing copper death according to claim 2, characterized in that: The pH is adjusted by a weak acid; The pH value is 4.0–6.0; The soluble inorganic tellurium salt is sodium tellurite.

4. The method for preparing tellurium-based nanomaterials with high efficiency inducing copper death according to claim 1, characterized in that: The amounts of the thiol-containing compound and the soluble copper salt mentioned in step (1) are calculated based on the molar ratio of sulfur in the thiol-containing compound to copper in the soluble copper salt being not less than 2:1; The Cu-S2 and the TeO2 nanocarrier mentioned in step (2) are mixed in a mass ratio of 75-85:1100-1200.

5. The method for preparing tellurium-based nanomaterials with high efficiency inducing copper death according to claim 4, characterized in that: The amounts of the thiol-containing compound and the soluble copper salt mentioned in step (1) are calculated based on a molar ratio of sulfur in the thiol-containing compound to copper in the soluble copper salt of 2.1 to 2.3:1; The Cu-S2 and the TeO2 nanocarrier mentioned in step (2) are mixed in a copper-tellurium mass ratio of 80:1120.

6. The method for preparing tellurium-based nanomaterials with high efficiency inducing copper death according to claim 1, characterized in that: The stirring speed mentioned in step (1) is 200-400 rpm; The stirring reaction time described in step (1) is 5 to 15 minutes; The stirring speed mentioned in step (2) is 200-400 rpm; The stirring reaction time in step (2) is 0.5 to 2 hours; The solid-liquid separation method described in step (2) is centrifugation.

7. A tellurium-based nanomaterial that efficiently induces copper death, characterized in that: It is obtained by the preparation method according to any one of claims 1 to 6.

8. The use of the tellurium-based nanomaterials that efficiently induce copper death as described in claim 7 in the preparation of X-ray radiotherapy sensitizers or antitumor drugs.

9. The application according to claim 8, characterized in that: The tumor in question is a cancer of the reproductive system.

10. The application according to claim 9, characterized in that: The reproductive system cancer mentioned is cervical cancer.

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