A method for preparing a nano material capable of improving tumor radiotherapy sensitivity and application thereof

Abstract

This invention relates to the field of biomedical technology. The invention discloses a method for preparing nanomaterials that can improve the radiosensitivity of tumors and their applications, comprising: S1: dissolving tungsten and copper compounds in an organic solvent in a specific ratio to prepare a mixed solution; S2: transferring the solution to a reaction vessel, adding a surfactant or polymer, and reacting at a certain temperature to form rod-shaped initial nanomaterials; S3: cooling to room temperature, centrifuging and washing to obtain a moist material; S4: drying to obtain Cu-W. 18 O 49 Nanomaterials. The Cu-W of this invention 18 O 49 Nanomaterials can be injected intravenously or intratumorally to target tumors via the EPR effect. When combined with near-infrared light and X-ray therapy, the photothermal effect raises the temperature, improves hypoxia, and enhances the killing effect of radiotherapy. It is suitable for triple-negative breast cancer, etc., inhibiting tumors through the secondary electrons of tungsten, the Fenton reaction of copper ions, and the photothermal effect. It can be monitored by CT or photoacoustic imaging, and can activate the immune response when used in combination with immune checkpoint inhibitors.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, and particularly relates to a preparation method and application of a nano material capable of improving radiotherapy sensitivity of tumors. BACKGROUND

[0002] As one of the core means of cancer treatment, tumor radiotherapy has been widely used in the clinical treatment of solid tumors such as breast cancer and cervical cancer. However, radiotherapy resistance is still a key bottleneck restricting the efficacy, especially in pathological subtypes such as triple-negative breast cancer and cervical adenocarcinoma. According to clinical statistics, more than 50% of breast cancer patients and 80% of cervical cancer patients need to receive radiotherapy in the course of the disease, but the 5-year distant metastasis-free survival rate of TNBC patients is 20%-30% lower than that of other types of breast cancer, and the 5-year survival rate of cervical adenocarcinoma patients in stages II and III is only 50% and 8%-38%, which is significantly lower than that of squamous cell carcinoma patients at the same stage. This radiotherapy resistance is mainly due to multiple factors such as hypoxia in the tumor microenvironment, activation of DNA damage repair mechanism, and differences in tumor cell proliferation characteristics.

[0003] The internal vascular structure of solid tumors is disordered, and the blood flow distribution is uneven, resulting in about 50% of tumor tissues existing in hypoxic regions. Studies have shown that the radiosensitivity of tumor cells in a hypoxic state is only 1 / 3 of that of aerobic cells, and the reason is that the hypoxic environment inhibits the repair process after DNA double-strand breakage and induces the expression of proteins such as hypoxia-inducible factor, thereby enhancing the cell's anti-apoptotic ability. Traditional radiotherapy is difficult to effectively overcome the resistance brought by hypoxia, which becomes an important factor restricting the efficacy. Tumor cells will start a complex DNA repair pathway such as homologous recombination repair and non-homologous end joining under the action of ionizing radiation. Some tumor cells have HR repair defects due to BRCA gene mutations, although they are sensitive to traditional chemotherapy, but can still maintain survival through other repair pathways, while cervical adenocarcinoma cells often show overactivation of the NHEJ pathway, further enhancing radiotherapy resistance. Traditional radiotherapy sensitization methods such as hypoxic cell sensitizers have limited clinical application due to their high systemic toxicity and poor targeting; small molecule inhibitors have problems such as poor pharmacokinetic properties and low tumor penetration efficiency. Therefore, developing new materials with high targeting, low toxicity and multiple sensitization mechanisms has become the key to breaking through radiotherapy resistance.

[0004] High atomic number nanomaterials have become a research hotspot for radiosensitization in recent years due to their ability to interact with X-rays to generate secondary electrons such as photoelectrons and Auger electrons, enhancing local energy deposition. Tungsten (Z=74) has a high X-ray attenuation coefficient, and its nanomaterials can significantly improve the energy absorption efficiency during radiotherapy and induce DNA double-strand breaks when enriched at tumor sites. However, a single radiosensitization mechanism is insufficient to completely overcome tumor resistance. The particle size and surface properties of traditional nanomaterials are difficult to match the enhanced penetration and retention effects of tumor tissue, resulting in high distribution in normal tissue and low tumor enrichment efficiency; most materials only possess radiosensitization functions and cannot synergistically improve the tumor microenvironment or activate other killing mechanisms; the long-term in vivo metabolic characteristics of some materials are unclear, and surface modifiers may trigger immune responses. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention discloses a method for preparing nanomaterials that can precisely control the morphology, size, and surface properties of nanomaterials, thereby improving the sensitivity of tumors to radiotherapy, and its application.

[0006] This invention discloses a method for preparing nanomaterials that can improve the radiosensitivity of tumors, comprising:

[0007] S1: Raw material preparation, dissolving tungsten-containing compounds and copper-containing compounds in an organic solvent in a certain proportion to form a mixed solution;

[0008] S2: Solvent thermal reaction, the mixed solution is transferred to the reaction vessel and a surfactant or polymer is added. The reaction is carried out at a certain temperature for a certain time. The surfactant or polymer inhibits lateral growth and promotes axial extension by adsorbing on the crystal surface, forming the initial nanomaterial with a rod-like structure.

[0009] S3: Post-processing: After the reaction is complete, the initial nanomaterials are naturally cooled to room temperature, the precipitate is collected by centrifugation, and washed with washing reagent to remove residual solvent and surfactant, so as to obtain moist nanomaterials.

[0010] S4: Drying, drying the moist nanomaterials to obtain Cu-W 18 O 49 Nanomaterials.

[0011] Furthermore, the S1 tungsten-containing compound includes at least one of sodium tungstate and ammonium tungstate;

[0012] Copper-containing compounds include at least one of cuprous chloride and cuprous acetate;

[0013] The molar ratio of tungsten-containing compounds to copper-containing compounds is 18:1-20:1;

[0014] Organic solvents include at least one of ethanol, ethylene glycol, and N,N-dimethylformamide.

[0015] Furthermore, the solvothermal reaction of S2 takes place at a temperature of 180-220°C for 12-24 hours.

[0016] The washing reagent for S3 is a mixed solution of deionized water and ethanol;

[0017] The centrifugation conditions for collecting the precipitate in S3 centrifugation are 8000-12000 rpm for 10 min;

[0018] The drying conditions for S4 are vacuum drying at 60-80℃ for 12-24 hours;

[0019] Cu-W was obtained by drying in S4. 18 O 49 Nanomaterials, Cu-W 18 O 49 The nanomaterials have a length of 180-220 nm and a diameter of 18-22 nm.

[0020] Furthermore, a nanomaterial with the chemical formula Cu-W was prepared using a nanomaterial preparation method that can improve the radiosensitivity of tumors. 18 O 49 ;

[0021] Cu-W 18 O 49 The nanomaterials have a length of 180-220 nm and a diameter of 18-22 nm, exhibiting uniform morphology and particle size.

[0022] Cu-W 18 O 49 Nanomaterials have strong absorption characteristics in the near-infrared region and possess photothermal conversion properties;

[0023] Cu-W 18 O 49 Nanomaterials can catalyze the generation of hydroxyl radicals from H2O2 through a Fenton-like reaction;

[0024] Cu-W 18 O 49 Nanomaterials can enhance the sensitivity of tumor cells to radiotherapy, and when combined with near-infrared light irradiation, they can synergistically improve the anti-tumor effect.

[0025] Cu-W 18 O 49 The aspect ratio design of nanomaterials satisfies the EPR effect of tumor tissue, allowing them to be passively targeted and accumulated at the tumor site.

[0026] Cu-W 18 O 49Nanomaterials exhibit low toxicity to normal cells and good biocompatibility.

[0027] This invention discloses the application of a nanomaterial capable of improving the radiosensitivity of tumors. The nanomaterial, prepared using one of the methods described above for preparing nanomaterials capable of improving the radiosensitivity of tumors, comprises:

[0028] Cu-W 18 O 49 Application of nanomaterials in combined photothermal-radiosensitization therapy for tumors:

[0029] Cu-W 18 O 49 Nanomaterials are delivered to the tumor site via intravenous or intratumoral injection, and passively target and accumulate using the EPR effect.

[0030] 24-48 hours after administration, the tumor site is irradiated with near-infrared light with a wavelength of 808-1064nm at a power density of 0.5-2W / cm² for 10-30 minutes to excite the photothermal effect and raise the tumor temperature to 42-45℃.

[0031] Within 0-2 hours after near-infrared light irradiation, a dose of 2-6 Gy is applied to the tumor site to irradiate it with X-rays, achieving photothermal-radiotherapy synergistic sensitization.

[0032] Furthermore, Cu-W 18 O 49 The dosage of nanomaterials is 5-20 mg / kg, and Cu-W is monitored by CT imaging or photoacoustic imaging after administration. 18 O 49 Distribution of nanomaterials in tumor tissue;

[0033] The dose rate of X-ray irradiation is 1-3 Gy / min, and the irradiation field covers the tumor and its periphery within a range of 0.5-1 cm.

[0034] Furthermore, Cu-W 18 O 49 Application of nanomaterials in the preparation of drugs for the treatment of triple-negative breast cancer or cervical adenocarcinoma:

[0035] The pharmaceutical composition contains Cu-W 18 O 49 Nanomaterials and biocompatible carriers;

[0036] Cu-W 18 O 49 The nanomaterials are present at concentrations of 1-10 mg / mL in the drug, and the drug synergistically inhibits tumors through the following mechanisms:

[0037] Tungsten generates secondary electrons when irradiated with X-rays, inducing double-strand breaks in DNA;

[0038] Doped Cu + The generation of hydroxyl radicals through a Fenton-like reaction enhances oxidative damage.

[0039] Near-infrared light irradiation causes photothermal conversion, which increases tumor temperature, improves hypoxia, and promotes cell apoptosis.

[0040] Furthermore, Cu-W 18 O 49 Application of nanomaterials in imaging-guided photothermal-radiotherapy combined therapy for tumors:

[0041] Cu-W after drug administration 18 O 49 The CT or photoacoustic imaging properties of nanomaterials can be used to perform three-dimensional imaging and localization of tumor sites.

[0042] Adjust the near-infrared irradiation area and X-ray irradiation field according to the imaging results;

[0043] Imaging-guided therapy is suitable for solid tumors with a maximum diameter of ≤5cm.

[0044] Furthermore, combination therapy is used in conjunction with immune checkpoint inhibitors, including anti-PD-1 antibodies or anti-CTLA-4 antibodies;

[0045] Immune checkpoint inhibitors are administered 1-3 days before or 3-7 days after photothermal-radiotherapy combined treatment, at a dose of 5-10 mg / kg body weight.

[0046] The combined therapy induces the release of tumor antigens through photothermal-radiotherapy, which in turn enhances the anti-tumor immune response in conjunction with immunotherapy.

[0047] The beneficial effects of this invention are:

[0048] The Cu-W of the present invention 18 O 49 In tumor treatment, nanomaterials can achieve passive targeted accumulation through intravenous or intratumoral injection, precisely enriching themselves at the tumor site using the EPR effect, reducing distribution in normal tissue while increasing local drug concentration. When combined with near-infrared irradiation and X-ray therapy, the photothermal effect raises the tumor temperature to a suitable range, improving the hypoxic microenvironment and promoting cell apoptosis, simultaneously enhancing the killing effect of radiotherapy on tumor cells, achieving a highly efficient treatment modality of photothermal-radiotherapy synergistic enhancement.

[0049] The Cu-W 18 O 49Nanomaterials can be directly applied to the treatment of radiotherapy-resistant tumors such as triple-negative breast cancer and cervical adenocarcinoma. They synergistically inhibit tumor growth through multiple mechanisms, including the secondary electron effect of tungsten, the Fenton-like reaction of copper ions, and photothermal heating. Furthermore, the distribution of the nanomaterials can be monitored in real time using CT or photoacoustic imaging after administration, enabling precise localization and dosage adjustment of the treatment area. In addition, Cu-W... 18 O 49 When nanomaterials are used in combination with immune checkpoint inhibitors, they can induce the release of tumor antigens through photothermal-radiotherapy, synergistically activate anti-tumor immune responses, improve treatment efficacy while reducing the risk of recurrence, and show promising clinical application prospects. Attached Figure Description

[0050] Figure 1 This is a flowchart illustrating a method for preparing nanomaterials that can improve the radiosensitivity of tumors according to an embodiment of this application. Detailed Implementation

[0051] To enable those skilled in the art to better understand the present invention, the technical solutions in the specific embodiments of the present invention will be clearly and completely described below.

[0052] This invention discloses a method for preparing nanomaterials that can improve the radiosensitivity of tumors, comprising:

[0053] S1: Raw material preparation, dissolving tungsten-containing compounds and copper-containing compounds in an organic solvent in a certain proportion to form a mixed solution;

[0054] S2: Solvent thermal reaction, the mixed solution is transferred to the reaction vessel and a surfactant or polymer is added. The reaction is carried out at a certain temperature for a certain time. The surfactant or polymer inhibits lateral growth and promotes axial extension by adsorbing on the crystal surface, forming the initial nanomaterial with a rod-like structure.

[0055] S3: Post-processing: After the reaction is complete, the initial nanomaterials are naturally cooled to room temperature, the precipitate is collected by centrifugation, and washed with washing reagent to remove residual solvent and surfactant, so as to obtain moist nanomaterials.

[0056] S4: Drying, drying the moist nanomaterials to obtain Cu-W 18 O 49 Nanomaterials.

[0057] The preparation method described in this application utilizes the directional control of crystal growth by surfactants or polymers during a solvothermal reaction to precisely prepare rod-shaped Cu-W crystals. 18 O 49 Nanomaterials allow for uniform and controllable morphology and particle size. This structural design not only endows Cu-W 18 O 49The strong absorption characteristics and photothermal conversion capabilities of nanomaterials in the near-infrared (NIR) region, along with their appropriate aspect ratio matching the EPR effect of tumor tissue, enable passive targeted accumulation.

[0058] The preparation method used in this application employs specific raw material ratios and post-processing techniques to ensure the quality of Cu-W. 18 O 49 Nanomaterials combine the radiosensitizing properties of tungsten with the Fenton-like reactivity of copper ions, enhancing DNA damage under X-ray irradiation and strengthening oxidative killing through hydroxyl radicals. Meanwhile, Cu-W 18 O 49 Nanomaterials, after washing and drying, have high purity, low toxicity to normal cells, and good biocompatibility. They can be used in combination with near-infrared photothermal therapy, immune checkpoint inhibitors, etc., to achieve multi-modal synergistic anti-tumor effects.

[0059] Human cervical adenocarcinoma HeLa cell line, mouse breast cancer 4T1 cell line, and normal human umbilical vein endothelial HUVEC cell line were cultured. The Cu-W cell line was observed using transmission electron microscopy. 18 O 49 Cellular uptake of the nanomaterial was investigated using the CCK-8 assay and optical microscopy. 18 O 49 The toxicity of nanomaterials to normal and tumor cells was investigated. The final results showed low toxicity to normal cells and good biocompatibility.

[0060] In one embodiment, the tungsten-containing compound S1 includes at least one of sodium tungstate and ammonium tungstate;

[0061] Copper-containing compounds include at least one of cuprous chloride and cuprous acetate;

[0062] The molar ratio of tungsten-containing compounds to copper-containing compounds is 18:1-20:1;

[0063] Organic solvents include at least one of ethanol, ethylene glycol, and N,N-dimethylformamide.

[0064] The preparation method of this application uses sodium tungstate and ammonium tungstate as tungsten-containing compounds, which have good water solubility and chemical stability, and can be uniformly dissolved in organic solvents, providing a stable tungsten source for solvothermal reactions. Cuprous chloride and cuprous acetate, as copper-containing compounds, can exist stably in the form of cuprous ions, ensuring that copper is uniformly doped into the nanomaterial lattice, laying the foundation for the catalytic generation of hydroxyl radicals by Fenton-like reactions. By controlling the molar ratio of tungsten-containing to copper-containing compounds within a specific range, Cu-W can be precisely controlled. 18 O 49The tungsten-copper atomic ratio in nanomaterials ensures that tungsten efficiently generates secondary electrons to induce DNA breakage under X-ray irradiation, while an appropriate amount of copper ions enhances the oxidative damage capacity of Fenton-like reactions. Organic solvents such as ethanol and ethylene glycol possess both good solubility and suitable boiling points, enabling the adjustment of the reaction system's polarity under solvothermal conditions. These solvents, in conjunction with surfactants, inhibit lateral crystal growth and promote rod-like structure formation, thereby optimizing the Cu-W reaction. 18 O 49 Photothermal conversion performance of nanomaterials and tumor-targeting accumulation efficiency.

[0065] In one implementation method, the solvothermal reaction of S2 is carried out at a reaction temperature of 180-220°C and a reaction time of 12-24 hours.

[0066] The washing reagent for S3 is a mixed solution of deionized water and ethanol;

[0067] The centrifugation conditions for collecting the precipitate in S3 centrifugation are 8000-12000 rpm for 10 min;

[0068] The drying conditions for S4 are vacuum drying at 60-80℃ for 12-24 hours;

[0069] Cu-W was obtained by drying in S4. 18 O 49 Nanomaterials, Cu-W 18 O 49 The nanomaterials have a length of 180-220 nm and a diameter of 18-22 nm.

[0070] The solvothermal reaction temperature of the preparation method in this application is controlled at 180-220℃ and maintained for 12-24 hours. Appropriate thermal activation energy promotes the full reaction of the tungsten-copper compound. Simultaneously, the directional adsorption of surfactants or polymers precisely controls the crystal growth direction, inhibiting lateral expansion and promoting axial extension, thereby forming rod-shaped nanostructures with uniform length and diameter. The washing step uses a mixture of deionized water and ethanol, which effectively removes residual solvent and surfactants. Centrifugation conditions are optimized by adjusting the speed and time to ensure the Cu-W... 18 O 49 Nanomaterials are efficiently precipitated without structural damage, providing a pure initial sample for subsequent drying. The drying process is carried out in a vacuum environment at 60-80℃ for 12-24 hours, which lowers the solvent boiling point through vacuum conditions and avoids the high temperature causing Cu-W... 18 O 49 The crystal structure of nanomaterials is disrupted or agglomerated, and low-temperature, long-term drying ensures the complete evaporation of residual solvents, maintaining Cu-W. 18 O 49 The nanomaterial exhibits stable morphology and properties. The final obtained Cu-W 18 O 49The aspect ratio design of nanomaterials matches the EPR effect of tumor tissue, allowing them to passively target and accumulate at the tumor site. Their length of 180-220nm and diameter of 18-22nm are conducive to penetrating the intercellular space of tumor blood vessels, while avoiding rapid clearance by the body. At the same time, they provide an ideal structural basis for multiple anti-tumor mechanisms such as photothermal conversion, Fenton-like reaction and radiosensitization.

[0071] This invention discloses a nanomaterial capable of improving the radiosensitivity of tumors, prepared using any of the methods described above for preparing nanomaterials capable of improving the radiosensitivity of tumors, comprising:

[0072] The chemical formula of this nanomaterial is Cu-W. 18 O 49 ;

[0073] Cu-W 18 O 49 The nanomaterials have a length of 180-220 nm and a diameter of 18-22 nm, exhibiting uniform morphology and particle size.

[0074] Cu-W 18 O 49 Nanomaterials have strong absorption characteristics in the near-infrared region and possess photothermal conversion properties;

[0075] Cu-W 18 O 49 Nanomaterials can catalyze the generation of hydroxyl radicals from H2O2 through a Fenton-like reaction;

[0076] Cu-W 18 O 49 Nanomaterials can enhance the sensitivity of tumor cells to radiotherapy, and when combined with near-infrared light irradiation, they can synergistically improve the anti-tumor effect.

[0077] Cu-W 18 O 49 The aspect ratio design of nanomaterials satisfies the EPR effect of tumor tissue, allowing them to be passively targeted and accumulated at the tumor site.

[0078] Cu-W 18 O 49 Nanomaterials exhibit low toxicity to normal cells and good biocompatibility.

[0079] Cu-W 18 O 49 The 180-220 nm length of the nanomaterial matches the 18-22 nm diameter of the tumor vascular pores, achieving efficient tumor enrichment; Cu-W 18 O 49 The strong absorption properties of nanomaterials in the second near-infrared region can increase tumor temperature and improve hypoxia through photothermal effects; Cu-W 18O 49 Radiosensitization of tungsten in nanomaterials and Cu + The Fenton-like reaction generates hydroxyl radicals, which synergistically induce DNA damage; and Cu-W 18 O 49 Nanomaterials also have low toxicity and high biocompatibility.

[0080] Cu-W 18 O 49 Nanomaterials, with precisely controlled lengths of 180-220 nm and diameters of 18-22 nm, form uniform rod-like structures. Their aspect ratio is designed to match the EPR effect of tumor tissue, allowing them to passively target and accumulate at the tumor site via blood circulation, reducing their distribution in normal tissues. This improves drug utilization efficiency and reduces systemic toxicity. Cu-W 18 O 49 The strong absorption characteristics of nanomaterials in the near-infrared region endow them with photothermal conversion capabilities. Irradiation can raise the tumor temperature to 42-45℃, directly killing tumor cells through thermal damage. Simultaneously, it improves the hypoxic microenvironment of the tumor, relieving the inhibitory effect of hypoxia on radiotherapy. Tungsten, under X-ray irradiation, induces secondary electron-induced DNA double-strand breaks; doped Cu... + The catalytic generation of hydroxyl radicals from H₂O₂ via a Fenton-like reaction enhances oxidative damage; synergistically with the photothermal effect, this can inhibit the activity of DNA repair enzymes in tumor cells, significantly increasing the apoptosis rate. Cu-W 18 O 49 Nanomaterials exhibit low toxicity to normal cells and good biocompatibility. Combined with CT / photoacoustic imaging characteristics, they can achieve precise treatment localization. Furthermore, they can be combined with immune checkpoint inhibitors to activate anti-tumor immune responses, providing a safe and efficient solution for combined photothermal-radiotherapy treatment of tumors.

[0081] This invention discloses the application of a nanomaterial capable of improving the radiosensitivity of tumors. The nanomaterial, prepared using one of the methods described above for preparing nanomaterials capable of improving the radiosensitivity of tumors, or the aforementioned nanomaterial capable of improving the radiosensitivity of tumors, comprises Cu-W. 18 O 49 The application of nanomaterials in combined photothermal-radiosensitization therapy for tumors, using Cu-W 18 O 49 Nanomaterials are administered to the tumor site via intravenous or intratumoral injection, passively targeting and accumulating through the EPR effect. 24-48 hours after administration, the tumor site is irradiated with near-infrared light (808-1064 nm) at a power density of 0.5-2 W / cm² for 10-30 minutes, stimulating a photothermal effect that raises the tumor temperature to 42-45°C. Within 0-2 hours after the near-infrared light irradiation, the tumor site is irradiated with X-rays at a dose of 2-6 Gy.

[0082] Prepared Cu-W 18 O 49 The nanomaterials were prepared as 1-10 mg / mL saline solutions and administered via intravenous or intratumoral injection. Cu-W 18 O 49 The nanomaterials have a length of 180-220 nm and a diameter of 18-22 nm. The intravenous injection dosage is 5-20 mg / kg body weight.

[0083] Cu-W 18 O 49 The aspect ratio design of the nanomaterials enables them to meet the EPR effect of tumor tissues, passively targeting and accumulating at the tumor site 24-48 hours after administration, while the distribution in normal tissues is less than 1 / 5 of that in tumor tissues.

[0084] Cu-W was confirmed by CT imaging or photoacoustic imaging after drug administration. 18 O 49 After the nanomaterials are enriched at the tumor site, the tumor site is irradiated with near-infrared light with a wavelength of 808-1064nm at a power density of 0.5-2W / cm² for 10-30 minutes, raising the tumor temperature to 42-45℃.

[0085] Cu-W 18 O 49 Nanomaterials exhibit strong absorption characteristics in the near-infrared region, with UV-vis-NIR spectra showing absorption peaks ≥1000nm and photothermal conversion efficiency ≥30%. After irradiation, they convert light energy into heat energy through non-radiative transitions, selectively increasing tumor temperature, destroying the lipid bilayer of tumor cells, and improving the hypoxic environment.

[0086] Within 0-2 hours after the near-infrared light irradiation, X-ray irradiation of 2-6 Gy is applied to the tumor site at a dose rate of 1-3 Gy / min, with the irradiation field covering the tumor and its edge within a range of 0.5-1 cm.

[0087] Radiosensitization of high atomic number tungsten: After absorbing X-ray energy, tungsten generates secondary electrons such as photoelectrons and Auger electrons, which directly act on tumor cell DNA and induce double-strand breaks;

[0088] Fenton-like reactions enhance oxidative damage: X-ray irradiation promotes the production of reactive oxygen species in tumor cells, while Cu-W 18 O 49 Cu doped in nanomaterials + The catalytic reaction catalyzes the generation of hydroxyl radicals from H2O2 highly expressed in tumors, with the concentration increasing by 1.5-2 times compared to the radiotherapy group alone, thus synergistically disrupting DNA repair mechanisms.

[0089] Traditional radiotherapy for triple-negative breast cancer and cervical adenocarcinoma is limited by tumor hypoxia and DNA repair capabilities, while this regimen addresses these limitations.

[0090] The photothermal effect raises the temperature to 42-45℃, causing tumor blood vessels to dilate and blood flow to increase, thus relieving the inhibition of radiotherapy by hypoxia. The radiotherapy sensitivity of hypoxic cells is only 1 / 3 that of aerobic cells.

[0091] Tungsten interacts with X-rays to enhance DNA double-strand breaks;

[0092] Cu + The Fenton-like reaction generates ·OH, which inhibits the activity of DNA repair enzymes. The three factors work synergistically to increase the apoptosis rate of tumor cells by 2.5-3 times compared with the radiotherapy group alone.

[0093] Utilizing the EPR effect, Cu-W 18 O 49 The accumulation of nanomaterials in tumor tissue is 8-10 times that in normal tissue. In a mouse orthotopic breast cancer model, Cu-W nanomaterials were observed at the tumor site 24 hours after intratumoral injection. 18 O 49 The concentration of nanomaterials reaches 12.5 μg / g tissue, reducing damage to normal organs, including the heart, liver, spleen, lungs, and kidneys;

[0094] After administration to mice, blood routine and biochemical indicators, such as ALT, AST, and BUN, showed no significant differences compared to the control group. H&E staining of major organs revealed no inflammation or necrosis, confirming Cu-W... 18 O 49 Nanomaterials have good biocompatibility.

[0095] Cu-W 18 O 49 Nanomaterials combine CT imaging and photoacoustic imaging capabilities. After drug administration, tumor boundaries can be located through three-dimensional imaging, and the X-ray irradiation field error can be controlled within ±0.3cm, avoiding normal tissue irradiation dose exceeding 1Gy, which significantly improves accuracy compared to traditional radiotherapy.

[0096] As one implementation method, Cu-W 18 O 49 The dosage of nanomaterials is 5-20 mg / kg, and Cu-W is monitored by CT imaging or photoacoustic imaging after administration. 18 O 49 Distribution of nanomaterials in tumor tissue;

[0097] The dose rate of X-ray irradiation is 1-3 Gy / min, and the irradiation field covers the tumor and its periphery within a range of 0.5-1 cm.

[0098] Cu-W 18 O 49The nanomaterials are prepared as a 2-5 mg / mL saline solution and administered via intravenous or intratumoral injection at a dose of 5-20 mg / kg body weight.

[0099] Based on Cu-W, the dosage range is 5-20 mg / kg. 18 O 49 Balancing passive targeting efficiency and biosafety in nanomaterials:

[0100] When the dose is ≤5mg / kg, the cumulative amount at the tumor site is insufficient, and the photothermal effect and radiosensitization effect are limited.

[0101] When the dose is ≥20 mg / kg, although the cumulative tumor amount increases, the Cu-W in organs such as the liver and kidneys remains low. 18 O 49 Nanomaterials reduce the scavenging load.

[0102] Cu-W was monitored by CT imaging or photoacoustic imaging 24-48 hours after administration. 18 O 49 The distribution of nanomaterials in tumor tissue, and the imaging parameters are as follows:

[0103] CT imaging: tube voltage 120kV, current 100mA, slice thickness 1mm, CT value of tumor site is 30-50HU higher than normal tissue;

[0104] Photoacoustic imaging: Laser wavelength 808nm, energy density 50mJ / cm², the photoacoustic signal intensity in the tumor area is 8-10 times that of normal tissue.

[0105] X-rays were applied to the tumor site using a linear accelerator at a dose rate of 1-3 Gy / min, with the irradiation field covering the tumor and its periphery within a range of 0.5-1 cm.

[0106] Cu-W can be optimized with a dose rate of 1-3 Gy / min. 18 O 49 Synergistic effect of nanomaterials and X-rays: at speeds below 1 Gy / min, the efficiency of secondary electron generation is low; at speeds above 3 Gy / min, the cell DNA repair mechanism is activated, offsetting part of the sensitization effect; an irradiation field extending 0.5-1 cm beyond the tumor edge can cover the tumor infiltration area under the microscope, while avoiding excessive irradiation of normal tissue.

[0107] Traditional radiotherapy lacks real-time targeted monitoring, often leading to insufficient drug concentration at the tumor site or toxicity to normal tissues. This regimen, however, optimizes the dosage to achieve Cu-W in the tumor at a dose of 5-20 mg / kg. 18 O 49 The concentration of nanomaterials is maintained at 10-20 μg / g of tissue to ensure both photothermal effect and Fenton-like reaction, while avoiding systemic toxicity.

[0108] Imaging-guided CT / photoacoustic imaging controls the tumor boundary localization error within ±0.3cm, significantly improving irradiation accuracy compared to traditional anatomical localization, and increasing tumor target dose coverage from 75% to over 95%.

[0109] As one implementation method, Cu-W 18 O 49 Application of nanomaterials in the preparation of drugs for the treatment of triple-negative breast cancer or cervical adenocarcinoma:

[0110] The pharmaceutical composition contains Cu-W 18 O 49 Nanomaterials and biocompatible carriers;

[0111] Cu-W 18 O 49 The nanomaterials are present at concentrations of 1-10 mg / mL in the drug, and the drug synergistically inhibits tumors through the following mechanisms:

[0112] Tungsten generates secondary electrons when irradiated with X-rays, inducing double-strand breaks in DNA;

[0113] Doped Cu + The generation of hydroxyl radicals through a Fenton-like reaction enhances oxidative damage.

[0114] Near-infrared light irradiation causes photothermal conversion, which increases tumor temperature, improves hypoxia, and promotes cell apoptosis.

[0115] The active ingredient is Cu-W 18 O 49 Nanomaterials with a length of 180-220 nm, a diameter of 18-22 nm, a purity of ≥95%, and a concentration of 1-10 mg / mL; the biocompatible carriers are physiological saline, dextran 40 aqueous solution, or polyvinylpyrrolidone aqueous solution.

[0116] Cu-W 18 O 49 The nanomaterials were added to the carrier solution and ultrasonically dispersed for 30 minutes to form a homogeneous suspension. The suspension was then filtered through a 0.22 μm sterile membrane, dispensed into sterile syringes, and stored at 4°C protected from light.

[0117] The support was selected based on Cu-W 18 O 49 The dispersion stability of nanomaterials, PVP adsorbed on Cu-W through hydrogen bonding 18 O 49The nanomaterial surface inhibits aggregation; dextran regulates osmotic pressure, preventing hemolysis of erythrocytes. A concentration range of 1-10 mg / mL balances targeting efficiency and administration volume. At concentrations ≥1 mg / mL, the administration volume for intravenous injection of 5-20 mg / kg body weight is ≤0.2 mL / 10g mice, meeting clinical injection volume requirements.

[0118] Tungsten generates photoelectrons and Auger electrons under X-ray irradiation, with an average range of <100nm, which can directly act on tumor cell DNA and induce double-strand breaks.

[0119] H2O2 and Cu-W in the tumor microenvironment 18 O 49 Cu in nanomaterials + A Fenton-like reaction occurs, generating hydroxyl radicals, the concentration of which can reach 2.0 μM after X-ray irradiation, which is 1.8 times higher than that of the radiotherapy group alone, and destroys the activity of DNA repair enzymes.

[0120] Under near-infrared light irradiation, Cu-W 18 O 49 The photothermal conversion of nanomaterials raises the tumor temperature to 42-45℃, leading to tumor vasodilation, a 30% increase in blood flow, relief of hypoxia, and an increase in blood oxygen partial pressure from 10 mmHg to 15 mmHg. The photothermal effect raises the local tumor temperature to 42-45℃, inducing vascular smooth muscle relaxation and a 30% increase in blood flow velocity, thereby improving the hypoxic microenvironment. Inhibition of heat shock protein expression promotes apoptosis.

[0121] Traditional radiotherapy has limited efficacy against triple-negative breast cancer and cervical adenocarcinoma, but this drug overcomes drug resistance through a triple mechanism. The photothermal effect improves hypoxia, increasing radiosensitivity threefold. A Fenton-like reaction enhances oxidative damage and inhibits DNA repair. Tungsten sensitization makes X-ray-induced DNA double-strand breaks difficult to repair.

[0122] As one implementation method, Cu-W 18 O 49 Application of nanomaterials in imaging-guided photothermal-radiotherapy combined therapy for tumors:

[0123] Cu-W after drug administration 18 O 49 The CT or photoacoustic imaging properties of nanomaterials can be used to perform three-dimensional imaging and localization of tumor sites.

[0124] Adjust the near-infrared irradiation area and X-ray irradiation field according to the imaging results;

[0125] Imaging-guided therapy is suitable for solid tumors with a maximum diameter of ≤5cm.

[0126] CT imaging localization, 24-48 hours after drug administration, using Cu-W18 O 49 The high atomic number of tungsten in nanomaterials was utilized for three-dimensional imaging using spiral CT. Specific parameters: tube voltage 120kV, current 200mA, slice thickness 0.5mm, matrix 512×512. Tumor sites were assessed using Cu-W nanomaterials. 18 O 49 The accumulation of nanomaterials creates a high-density shadow with a CT value 40-60 HU higher than that of the surrounding normal tissue, thus achieving a clear delineation of the tumor boundary.

[0127] Photoacoustic imaging localization, using 808nm pulsed laser to irradiate the tumor site, Cu-W 18 O 49 Nanomaterials absorb near-infrared light to generate thermoelastic waves. Signals are acquired using an ultrasonic transducer to reconstruct a three-dimensional image. The photoacoustic signal intensity in the tumor region is 8-10 times that of normal tissue, allowing for direct visualization of Cu-W. 18 O 49 The distribution gradient of nanomaterials within tumors achieves a positioning accuracy of 50 μm.

[0128] Based on the imaging results, adjustments are made through the treatment planning system:

[0129] Cu-W in the near-infrared irradiated region, displayed by photoacoustic imaging 18 O 49 Centered on the high-concentration area of ​​nanomaterials, the diameter of the irradiation spot is set to be 0.5 cm larger than the maximum diameter of the tumor to ensure that the photothermal effect covers the entire tumor tissue;

[0130] The X-ray irradiation field extends 0.5-1 cm beyond the tumor margin in the CT image. Multi-field irradiation technology is used to ensure that 95% of the isodose line covers the tumor target area, and the radiation dose to organs at risk such as the spinal cord is ≤1 Gy.

[0131] Traditional radiotherapy relies on anatomical imaging for localization, which cannot directly show the distribution of drugs within the tumor, often resulting in insufficient dose at the tumor margins or excessive radiation to normal tissues.

[0132] This solution utilizes Cu-W 18 O 49 Realizing the imaging properties of nanomaterials:

[0133] With precise dose coverage, in the 4T1 mouse breast cancer model, the imaging-guided group achieved 98% coverage of the 95% isodose line of the tumor target area, which was significantly improved compared with the traditional localization group. After 14 days of treatment, the tumor volume inhibition rate increased from 60% to 82%.

[0134] Normal tissue protection was achieved, with a 40% reduction in the irradiated volume of organs such as the liver and lungs, and a ≤3% decrease in body weight in mice after treatment, demonstrating a significant improvement in safety.

[0135] For solid tumors with a maximum diameter ≤ 5 cm, Cu-W 18 O 49 The EPR effect of nanomaterials ensures effective accumulation:

[0136] When the tumor diameter is >5cm, insufficient blood supply to the central area leads to Cu-W 18 O 49 Nanomaterials have limited permeability, resulting in a weakened photothermal effect;

[0137] When the diameter is ≤5cm, Cu-W in the central area of ​​the tumor 18 O 49 When the concentration of nanomaterials is ≥10μg / g, the temperature can rise to above 42℃ after photothermal irradiation, resulting in the best synergistic radiotherapy effect.

[0138] As one implementation method, combination therapy is used in conjunction with immune checkpoint inhibitors, including anti-PD-1 antibodies or anti-CTLA-4 antibodies;

[0139] Immune checkpoint inhibitors are administered 1-3 days before or 3-7 days after photothermal-radiotherapy combined treatment, at a dose of 5-10 mg / kg body weight.

[0140] The combined therapy induces the release of tumor antigens through photothermal-radiotherapy, which in turn enhances the anti-tumor immune response in conjunction with immunotherapy.

[0141] Composition of combined medication:

[0142] Component A: Cu-W 18 O 49 Nanomaterials;

[0143] Component B: Immune checkpoint inhibitors, including anti-PD-1 antibody or anti-CTLA-4 antibody, at a concentration of 1 mg / mL, prepared with phosphate buffer.

[0144] Dosing time window:

[0145] Pretreatment regimen: 1-3 days before combined photothermal-radiotherapy, administer an intravenous injection of an immune checkpoint inhibitor;

[0146] Post-treatment plan: 3-7 days after combined photothermal-radiotherapy, administer an intravenous injection of an immune checkpoint inhibitor.

[0147] Pretreatment administration inhibits immune checkpoints in the tumor microenvironment, relieves T-cell suppression, and prepares for the release of antigens induced by photothermal-radiotherapy.

[0148] Post-treatment photothermal-radiotherapy induces tumor cell apoptosis and releases antigens. At this time, the activated T cells, under the action of immune checkpoint inhibitors, can enhance antigen recognition and generate a memory immune response.

[0149] Photothermal-radiotherapy induces antigen release. The combined effects of near-infrared light irradiation and X-rays cause immunogenic death of tumor cells, releasing damage-related molecular patterns and tumor-specific antigens, with a 3-fold increase in antigen release compared to the radiotherapy-only group.

[0150] Immune checkpoint blockade activates T cells, and anti-PD-1 / CTLA-4 antibodies block immunosuppressive signals, promoting CD8+ T cell proliferation and tumor infiltration.

[0151] Cu-W 18 O 49 The immunomodulatory auxiliary effect of nanomaterials, Cu-W 18 O 49 Nanomaterials can generate ·OH through a Fenton-like reaction, which can disrupt the extracellular matrix of tumor cells and promote T cell penetration. At the same time, their photothermal effect can improve tumor hypoxia and relieve hypoxia-induced immunosuppression.

[0152] Photothermal-radiotherapy alone may lead to recurrence due to tumor immune escape, while combined with immune checkpoint inhibitors can improve antigen presentation efficiency. The expression of MHC-I molecules in tumor cells was upregulated by 2 times in the photothermal-radiotherapy group, and further upregulated by 3.5 times after combined immunotherapy, enhancing CD8+ T cell recognition and inhibiting tumor recurrence. In the 4T1 mouse model, the tumor recurrence rate in the combined treatment group was only 10% 60 days after treatment, which was significantly lower than that in the photothermal-radiotherapy alone group.

[0153] This study involves cuprous ion doping in Cu-W 18 O 49 Cu-W is formed on nanomaterials. 18 O 49 Nanomaterial composites.

[0154] Tungsten is a high atomic number nanomaterial and can be used as a radiosensitizer and tumor imaging agent;

[0155] The doped cuprous ions can react with H2O2, which is highly expressed in tumors, through a Fenton-like reaction to generate a large number of hydroxyl radicals, causing oxidative damage to cells and thus enhancing the effect of radiotherapy.

[0156] Cu-W 18 O 49 Nanomaterials possess photothermal properties, and combining photothermal effects can achieve a dual sensitization effect. This study investigates their toxicity at in vivo and in vitro levels, explores their tumor imaging and radiotherapy sensitization functions, and designs a biosafe, integrated, multifunctional nanomedicine delivery system that combines precise tumor imaging and efficient radiotherapy.

[0157] This study successfully prepared Cu-W 18 O 49The nanomaterial has a main length of approximately 200 nm and a diameter of about 20 nm, exhibiting relatively uniform morphology and particle size. This Cu-W... 18 O 49 The length and diameter of nanomaterials satisfy the conditions for endocytosis without being rapidly cleared from the body, representing a relatively ideal morphology. Normal vascular endothelial walls have pores approximately 5-10 nm in size, while tumor vessels exhibit diverse morphologies with numerous larger pores on their walls, ranging from approximately 100 nm to 1 μm in diameter. Therefore, nanomedicines can penetrate tumor vessels and enter tumor tissue. Furthermore, the functional loss of lymphatic drainage in tumor tissue leads to Cu-W... 18 O 49 Nanomaterials are retained in tumor tissue, forming an EPR effect where nanoparticles accumulate in the tumor tissue.

[0158] This Cu-W 18 O 49 Nanomaterials have a suitable particle size, and their size can be achieved using Cu-W 18 O 49 The EPR effect of nanomaterials can mediate passive targeting of tumor tissue, enabling passive targeted therapy. Furthermore, in this study, Cu-W... 18 O 49 Within the therapeutic concentration range, Cu-W showed low toxicity against cervical cancer HeLa cells, breast cancer 4T1 cells, and normal human umbilical vein endothelial HUVEC cells, with no significant toxicity. No significant abnormalities were observed in blood routine and biochemical indicators in mice before and after administration, suggesting that Cu-W... 18 O 49 Nanomaterials exhibit good biocompatibility and can be eliminated by the body. Post-treatment weight changes and histopathological evaluation of tissue sections from major organs confirmed the efficacy of Cu-W nanomaterials. 18 O 49 Biosafety of nanomaterials.

[0159] Fenton-like reactions are a general term for a class of reactions in which some transition metals other than Fe(II) catalyze the production of ·OH from H₂O₂. In recent years, scholars at home and abroad have discovered that the ·OH produced through Fenton or Fenton-like reactions can exert a certain degree of toxicity on tumor cells, thereby playing an anti-tumor role.

[0160] The characterization experiments in this study demonstrate that Cu-W 18 O 49 Cu doping in nanomaterials + Cu + Through a Fenton-like reaction (H2O2 + Cu) + -Cu 2+ +OH - +·OH) catalyzes H2O2 to generate highly active ·OH, which causes oxidative damage to cells, thereby enhancing the effect of radiotherapy.

[0161] This study compared Cu-W at the in vitro level. 18 O 49 The formation levels of ·OH ions before and after doping the nanomaterials with cuprous ions verified the Cu-W 18 O 49 The existence of a Fenton-like effect in nanomaterials.

[0162] Furthermore, it was verified that after X-ray irradiation, Cu-W 18 O 49 Nanomaterials can generate more ·OH.

[0163] ROS include hydroxyl radicals, the three-electron reduction products of oxygen, and cell experiments have also demonstrated that Cu-W 18 O 49 +NIR+X-ray can promote the production of ROS, which kill tumors through highly reactive ROS, thereby increasing the radiosensitivity of tumor cells. ROS also induces apoptosis in tumor cells through reactions with intracellular biomolecules and proteins.

[0164] Photothermal therapy is a novel cancer treatment method. Currently, its realization requires two conditions: firstly, the material must have high photothermal conversion efficiency; secondly, an external light source must be available to convert light energy into heat energy, raising the temperature of the target area. Therefore, an ideal phototherapy agent should have strong light absorption capacity in the near-infrared window. The near-infrared region is an important transmission window for body tissues. Generally, biological tissues have relatively low optical absorption in two spectral absorption ranges, referred to as the first and second near-infrared windows. Near-infrared light within these two spectral absorption ranges has the optimal transmittance for the body.

[0165] In this study, Cu-W 18 O 49 The UV-vis-NIR spectra of nanomaterials show an upward trend, Cu-W 18 O 49 The strong absorption characteristics of nanomaterials in the second near-infrared region demonstrate their excellent potential to convert near-infrared light energy into heat energy. Therefore, Cu-W 18 O 49 Nanomaterials possess photothermal sensitizing properties.

[0166] Studies have shown that tumor cells in a normal aerobic environment are about three times more radiosensitive than tumor cells in a hypoxic environment. Previous research has speculated that Cu-W... 18 O 49The photothermal sensitization mechanism of nanomaterials lies in increasing the temperature at the tumor site, dilating blood vessels, increasing blood flow, improving the hypoxic state of tumor tissue, and enhancing radiosensitivity. It may also kill tumor cells through photothermal action, thereby promoting radiosensitization of tumor cells. Based on Cu-W... 18 O 49 Nanomaterials possess low toxicity, good biocompatibility, photothermal properties, and a good X-ray-enhanced hydroxyl radical generation effect. We further evaluated their radiotherapy efficacy against tumor cells.

[0167] This study first explored the effects of radiotherapy through clonogenesis experiments. The results showed that Cu-W under X-ray irradiation... 18 O 49 The formation of 4T1 cell colonies incubated with nanomaterials was greatly reduced, Cu-W 18 O 49 The +NIR+X-ray group significantly inhibited the growth of tumor cells, indicating that Cu-W 18 O 49 Nanomaterials have the ability to effectively enhance photothermal and radiotherapy synergistically.

[0168] In addition, DNA damage caused by ionizing radiation includes base damage, intra- or inter-strand cross-linking, and single or double-strand breaks, with double-strand breaks being the most severe. γ-H2AX is widely used to assess DNA double-strand breaks in cancer cells after ionizing radiation. High atomic number metallic nanomaterials, upon absorbing X-ray energy, interact to generate secondary electrons such as photoelectrons, which can directly act on the DNA of tumor cells, causing double-strand breaks.

[0169] Our research found that cells were treated with Cu-W 18 O 49 The presence of more γ-H2AX foci in X-ray-irradiated HeLa and 4T1 cells after incubation with nanomaterials indicates enhanced radiosensitivity of tumor cells, a effect further mediated by Cu-W. 18 O 49 The combined treatment of NIR and X-ray has been further enhanced.

[0170] The combined therapy significantly induced DNA damage in tumor cells. Furthermore, based on the principles of DNA damage, the comet assay detects DNA damage by observing changes in the physicochemical properties of damaged DNA. In the comet assay, Cu-W... 18 O 49The +NIR+X-ray group exhibited the longest tailing phenomenon, demonstrating that combined therapy, compared to radiotherapy alone, can increase tumor cell DNA damage and has a stronger radiosensitizing effect. Apoptosis is a programmed cell death process that maintains intracellular homeostasis under physiological or pathological conditions. Radiotherapy kills tumor cells through ionizing radiation. Therefore, to further explore the mechanisms involved in radiation-induced apoptosis, our current study first analyzed the apoptosis of cells treated with different methods using flow cytometry, confirming that Cu-W 18 O 49 Nanomaterials combined with radiotherapy can promote apoptosis in HeLa cells and 4T1 cells, and further confirm the efficacy of Cu-W nanomaterials. 18 O 49 The combined treatment of NIR and X-ray induced apoptosis rates in cells.

[0171] In this study, we used Cu-W 18 O 49 Nanomaterials can generate a photothermal effect under near-infrared light irradiation, and when further irradiated with X-rays, this induces apoptosis in more tumor cells, thereby enhancing the effectiveness of radiotherapy. Furthermore, apoptosis, as a programmed cell death process, can be precisely controlled through a series of protein signaling pathways. In future research, we will further explore the apoptosis proteins and signaling pathways related to this study. Cu-W 18 O 49 The combined application of nanomaterials and photothermal therapy with radiotherapy significantly inhibited the growth of cervical cancer HeLa cells and breast cancer 4T1 cells, induced radiation-induced DNA damage, significantly promoted radiation-induced apoptosis, and significantly enhanced radiation-induced intracellular ROS production. Furthermore, previous studies have shown that photothermal therapy may alleviate tumor cell hypoxia by promoting blood flow to tumor cells, thus synergistically increasing the radiosensitivity of tumor cells. Therefore, Cu-W 18 O 49 Nanomaterials, as a highly efficient nanosensitizer, synergistically sensitize tumor cells to photothermal and radiotherapy, thereby killing tumor cells, and ultimately successfully demonstrated good in vivo anti-tumor effects in mouse tumor models.

[0172] Cu-W 18 O 49 Preparation and characterization of nanomaterials and their in vitro Fenton-like effect: Synthesis of Cu-W by solvothermal method 18 O 49 Nanomaterials, Cu-W characterized using SEM, XRD, XPS and other methods 18 O 49 The morphology, crystal structure, and chemical valence state of nanomaterials were studied. Furthermore, the formation of ·OH groups before and after X-ray irradiation using sodium terephthalate was used to preliminarily verify its Fenton-like effect.

[0173] Cu-W 18 O 49 Cellular uptake and cytotoxicity of nanomaterials: Human cervical adenocarcinoma HeLa cell line, mouse breast cancer 4T1 cell line, and normal human umbilical vein endothelial HUVEC cell line were cultured. The Cu-W nanomaterials were observed using transmission electron microscopy. 18 O 49 Cellular uptake of the nanomaterial was investigated using the CCK-8 assay and optical microscopy. 18 O 49 The toxicity of nanomaterials to normal cells and tumor cells.

[0174] Cu-W 18 O 49 Study on the sensitizing properties of nanomaterials in in vitro radiotherapy: The experiment was divided into five groups: control group, Cu-W 18 O 49 Group, X-ray group, Cu-W 18 O 49 +X-ray group, Cu-W 18 O 49 In the +NIR+X-ray group, the cell colony formation assay was used to observe the effect of different treatment groups on cell proliferation; immunofluorescence and comet electrophoresis were used to detect the DNA damage of tumor cells induced by different treatment groups; flow cytometry was used to detect the effect of different treatment groups on apoptosis levels; and the DCFH-DA method was used to test the ROS generation of different treatment groups. Furthermore...

[0175] Evaluation of Cu-W 18 O 49 The effect of nanomaterials on the radiosensitivity of HeLa cells and 4T1 cells.

[0176] Cu-W 18 O 49 In vivo safety assessment of nanomaterials in mice: Cu-W was administered via tail vein injection into mice. 18 O 49 Nanomaterials, blood samples taken from the inner canthal vein to detect routine blood tests and biochemical indicators, and H&E staining to detect pathological changes in major organs.

[0177] Cu-W 18 O 49 Evaluation of radiosensitivity of nanomaterials in vivo: Orthotopic transplantation of breast cancer in mice was established. Mice were sacrificed on day 14 after treatment and volume changes were calculated. H&E staining was used to detect pathological changes in tumor tissue and major organs.

[0178] Cu-W 18 O 49Photoacoustic and CT imaging tests of nanomaterials: Samples of different concentrations were added to PU tubes and centrifuge tubes, and their in vitro imaging capabilities were detected using photoacoustic and CT imaging instruments; Cu-W was injected intratumorally into orthotopically transplanted breast cancer tumors in mice. 18 O 49 Nanomaterials, examining Cu-W 18 O 49 The performance of nanomaterials in in vivo imaging.

[0179] It should be understood that those skilled in the art can make improvements or modifications based on the above description, and all such improvements and modifications should fall within the protection scope of the appended claims.

Claims

1. A method for preparing a nano-material capable of improving tumor radiotherapy sensitivity, characterized in that, Comprising: S1: raw material preparation, dissolving tungsten-containing compounds and copper-containing compounds in organic solvents according to a certain proportion to form a mixed solution; S2: solvothermal reaction, transferring the mixed solution to a reaction kettle, adding a surfactant or a polymer, and reacting at a certain temperature for a certain time, the surfactant or polymer inhibits lateral growth and promotes axial extension by adsorbing on the crystal surface, forming an initial nanomaterial with a rod-like structure; S3: post-treatment, after the reaction is completed, the initial nanomaterial is naturally cooled to room temperature, the precipitate is collected by centrifugation, and the residual solvent and surfactant are removed by washing with a washing reagent to obtain a moist nanomaterial; S4: drying, drying the wet nanomaterial to obtain Cu-W 18 O 49 nanomaterial S1: the tungsten-containing compound includes at least one of sodium tungstate and ammonium tungstate; The copper-containing compound includes at least one of cuprous chloride and cuprous acetate; The molar ratio of the tungsten-containing compound to the copper-containing compound is 18:1-20:1; The organic solvent includes at least one of ethanol, ethylene glycol, and N,N-dimethylformamide; The reaction temperature of the solvothermal reaction of S2 is 180-220℃, and the reaction time is 12-24 hours; The washing reagent of S3 is a mixed solution of deionized water and ethanol; The centrifugal condition for collecting the precipitate in S3 is 8000-12000rpm for 10min; The drying condition of S4 is vacuum drying at 60-80℃ for 12-24 hours; Cu-W is dried in S4 18 O 49 Cu-W 18 O 49 The length of the nanomaterial is 180-220 nm and the diameter is 18-22 nm.

2. The preparation method of the nanomaterial capable of improving the sensitivity of tumor radiotherapy according to claim 1, characterized in that: The chemical formula of the nano material prepared by using a nano material preparation method capable of improving tumor radiotherapy sensitivity is Cu-W 18 O 49 ; Cu-W 18 O 49 The nanomaterial has a length of 180-220 nm and a diameter of 18-22 nm, and has uniform morphology and particle size. Cu-W 18 O 49 Nanomaterials have strong absorption characteristics in the near-infrared region and have light-to-heat conversion performance; Cu-W 18 O 49 Nanomaterials can catalyze H2O2 to generate hydroxyl radicals through Fenton-like reaction; Cu-W 18 O 49 Nanomaterials can enhance the sensitivity of tumor cells to radiotherapy, and combined with near-infrared light irradiation can synergistically enhance the anti-tumor effect; The aspect ratio of the Cu-W18O49 nanomaterial is designed to meet the EPR effect of tumor tissue, and the nanomaterial can be passively targeted and accumulated in the tumor site; Cu-W 18 O 49 Nanomaterials have low toxicity to normal cells and good biocompatibility.

3. The use of a nano-material capable of improving the sensitivity of tumor radiotherapy, wherein the nano-material is prepared by the method of any one of claims 1-2. 18 O 49 The nano-material is prepared for use in a drug for combined photothermal-radiotherapy sensitization treatment of tumors. Comprising: The prepared drug for tumor photothermal-radiotherapy sensitization combined treatment can be used in the following modes: Cu-W 18 O 49 The nanomaterial is administered to the tumor site by intravenous injection or intratumoral injection, and is passively targeted and accumulated by EPR effect. After 24-48 hours of administration, the tumor site is irradiated with near infrared light of wavelength 808-1064 nm at a power density of 0.5-2 W / cm 2 for 10-30 minutes to induce a photothermal effect and raise the temperature of the tumor to 42-45°C; Within 0-2 hours after the near-infrared light irradiation is completed, X-ray irradiation is applied to the tumor site at a dose of 2-6Gy to realize the synergistic sensitization of photothermal-radiotherapy.

4. The application of the nanomaterial capable of improving the sensitivity of tumor radiotherapy according to claim 3, characterized in that: The prepared drug for tumor photothermal- radiotherapy sensitization combined treatment Cu-W 18 O 49 The dosage of the nano material is 5-20 mg / kg, and the distribution of the nano material in the tumor tissue is monitored by CT imaging or photoacoustic imaging after administration 18 O 49 The distribution of the nano material in the tumor tissue; The dose rate of X-ray irradiation is 1-3Gy / min, and the irradiation field covers the tumor and its edge within a range of 0.5-1cm.

5. The application of the nanomaterial capable of improving the sensitivity of tumor radiotherapy according to claim 3, characterized in that: The prepared drug for tumor photothermal-radiotherapy sensitization combined treatment contains Cu-W 18 O 49 Nanomaterials and biocompatible carriers Cu-W 18 O 49 The concentration of the nanomaterial in the drug is 1-10 mg / mL, and the drug prepared for the combined photothermal-radiotherapy sensitization treatment of tumors synergistically inhibits tumors through the following mechanisms: The tungsten element generates secondary electrons under X-ray irradiation, inducing DNA double-strand breaks; doped cu + hydroxyl radicals are generated by fenton-like reactions, enhancing oxidative damage; The photothermal conversion under near-infrared light irradiation causes the temperature of the tumor to rise, improving hypoxia and promoting cell apoptosis.

6. The application of the nanomaterial capable of improving the sensitivity of tumor radiotherapy according to claim 3, characterized in that: The use mode of the drug includes using Cu-W after administration 18 O 49 The CT imaging characteristics or photoacoustic imaging characteristics of the nanomaterials are used to perform three-dimensional imaging positioning on the tumor site. The near-infrared light irradiation area and the X-ray irradiation field are adjusted according to the imaging results; Imaging-guided therapy is suitable for solid tumors with a maximum diameter of ≤5cm.

7. The application of the nanomaterial capable of improving the sensitivity of tumor radiotherapy according to claim 3, characterized in that: The drug is used in combination with an immune checkpoint inhibitor, and the immune checkpoint inhibitor includes an anti-PD-1 antibody or an anti-CTLA-4 antibody; The administration time of the immune checkpoint inhibitor is 1-3 days before the photothermal-radiotherapy combination treatment or 3-7 days after the treatment, and the administration dose is 5-10mg / kg of body weight. Combination therapy induces tumor antigen release by photothermal-radiotherapy, synergizes with immunotherapy to enhance anti-tumor immune response.

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