High-entropy sub-nanowire compound for catalytic treatment of tumors as well as preparation method and application of high-entropy sub-nanowire compound

Abstract

The invention relates to the technical field of biomedical materials, and discloses a high-entropy sub-nanowire compound for catalytic treatment of tumors as well as a preparation method and application of the high-entropy sub-nanowire compound, and the high-entropy sub-nanowire compound comprises a PtFeMoCoNiRu high-entropy sub-nanowire and aldehyde polyether F127 modified on the surface of the PtFeMoCoNiRu high-entropy sub-nanowire. The high-entropy sub-nanowire compound is good in water solubility and high in biocompatibility; under ultrasonic stimulation, enzyme activity of oxidase-like enzyme, peroxidase and catalase can be further enhanced so as to generate various ROS (reactive oxygen species), rising GSH (glutathione) in a tumor microenvironment can be consumed at the same time, anti-oxidation capacity of tumor cells is reduced, and efficient anti-tumor performance is realized.

Description

Technical Field

[0001] This invention relates to the technical field of biomedical materials, and in particular to a high-entropy subnanowire composite for tumor catalytic therapy, its preparation method, and its application. Background Technology

[0002] Cancer treatment remains one of the major challenges facing the medical field today; with the continuous advancement of medical technology, cancer treatment methods are also constantly evolving and innovating. Different treatment methods all have certain limitations. For example, traditional surgery carries a significant risk of intraoperative bleeding and postoperative infection, while radiotherapy and chemotherapy cause considerable damage to surrounding normal tissues, leading to a series of adverse reactions. Therefore, cancer treatment still requires the exploration of new treatment methods to provide patients with the most efficient and safe treatment options.

[0003] The application of nanomaterials in cancer treatment has attracted widespread attention, especially biocatalytic materials that generate reactive oxygen species (ROS) for tumor therapy. Biocatalysts utilize excess H₂O₂ in the tumor microenvironment to activate multiple enzymatic reactions, generating ROS that kill tumor cells. Biocatalysts are central to the therapeutic effect, determining the pathway, selectivity, and overall therapeutic efficiency of the biocatalytic reaction. Although nanocatalytic materials have expanded to include noble metals, carbon materials, metal oxides, and metal-organic frameworks, they still suffer from structural instability in biological environments and insufficient catalytic efficiency, making it difficult to meet practical application requirements.

[0004] High entropy alloys (HEAs) are a class of materials with broad potential for catalytic applications. Through characteristics such as high entropy effect, lattice distortion, slow diffusion effect and "cocktail effect", they significantly improve the thermal stability of materials, optimize reaction energy barriers, and create diverse adsorption sites suitable for multi-step tandem reactions. The electronic hybridization effect between constituent elements forms quasi-continuous d-bands and generates unique active sites widely distributed on the material surface, thereby expanding the design space for regulating catalytic activity and selectivity. Summary of the Invention

[0005] To address the current situation where existing biocatalytic materials suffer from structural instability and insufficient catalytic efficiency in biological environments, this invention provides a PtFeMoCoNiRu-F high-entropy subnanowire complex (HESNW-F) with multiple enzymatic activities suitable for tumor catalytic therapy, along with its preparation method and applications. This high-entropy subnanowire complex exhibits good stability and biocompatibility. Under ultrasonic stimulation, it can further enhance the activities of oxidase-like enzymes (OXD), peroxidase (POD), and catalase (CAT) to generate various ROS, and simultaneously consume the elevated glutathione (GSH) in the tumor microenvironment, reducing the antioxidant capacity of tumor cells and achieving highly efficient anti-tumor performance.

[0006] In a first aspect, the present invention provides a high-entropy subnanowire composite, which adopts the following technical solution: A high-entropy subnanowire composite comprising PtFeMoCoNiRu high-entropy subnanowires and aldehyde-modified polyether F127 modified on the surface of the PtFeMoCoNiRu high-entropy subnanowires.

[0007] Optionally, the high-entropy subnanowire composite has a width ≤1 nm and a lattice width of 0.2 nm.

[0008] Optionally, the proportions of Pt, Fe, Mo, Co, Ni, and Ru atoms in the high-entropy subnanowire composite are all 5%-35%.

[0009] Optionally, the atomic ratio of Pt, Fe, Mo, Co, Ni and Ru in the high-entropy subnanowire composite is 33.8:21.7:22.5:9.0:7.8:5.12.

[0010] Optionally, the mass ratio of the aldehyde-modified polyether F127 to the high-entropy subnanowire is (4-5):1.

[0011] By adopting the above technical solution, the alloying of the PtFeMoCoNiRu-F high-entropy subnanowire composite caused its d-band center to shift significantly downward. Through the charge redistribution of multiple metals and the strong multi-orbital hybridization effect between alloying elements, the catalytic effect was significantly improved. Furthermore, F127 was used to significantly increase its biocompatibility, resulting in a novel and highly efficient tumor catalytic therapy material.

[0012] Secondly, the present invention provides a method for preparing a high-entropy subnanowire composite, employing the following technical solution: A method for preparing a high-entropy subnanowire composite includes the following steps: S1: Platinum acetylacetonate, iron acetylacetonate, molybdenum hexacarbonyl, cobalt acetylacetonate, nickel acetylacetonate, and ruthenium acetylacetonate were added to oleylamine and mixed at room temperature to obtain a first mixture; the first mixture was heated to react, cooled, centrifuged, and then washed with a mixed solvent of cyclohexane and ethanol to obtain high-entropy subnanowires; S2: Mix high-entropy subnanowires with aldehyde-modified polyether F127 evenly to obtain a second mixture; evaporate, dissolve, and wash the second mixture to obtain the final product.

[0013] Optionally, in step S1, the molar ratio of platinum acetylacetone, iron acetylacetone, molybdenum hexacarbonyl, cobalt acetylacetone, nickel acetylacetone, and ruthenium acetylacetone is 0.25:0.20:0.38:0.27:0.15:0.13.

[0014] Optionally, in step S2, the mass ratio of the aldehyde-modified polyether F127 to the high-entropy subnanowire is (4-5):1.

[0015] Thirdly, the present invention provides the application of a high-entropy subnanowire composite in the preparation of a drug for tumor catalytic therapy.

[0016] Optionally, the tumor catalytic therapy is performed in combination with ultrasound therapy.

[0017] Optionally, the drug is administered via intratumoral injection.

[0018] By employing the above-mentioned technical solution, under the action of ultrasound, electron transfer in high-entropy sub-nanowire composite materials is promoted, which significantly enhances the activity of various enzymes in nanomaterials, significantly promotes the generation of ROS and the consumption of GSH, and further improves the therapeutic effect of tumor treatment.

[0019] The beneficial effects of the technical solution of this invention are as follows: 1. This invention provides a high-entropy subnanowire composite for tumor catalytic therapy, which is obtained by modifying high-entropy subnanowires with aldehyde-modified polyether F127; the high-entropy material with subnanostructure exposes more active sites, and the surface-modified aldehyde-modified polyether F127 improves water solubility and biocompatibility; 2. The high-entropy sub-nanowire composite of the present invention can be used for catalytic therapy of tumors by intratumoral injection. It has a long residence time at the tumor site, can generate more ROS in the tumor area, enhance the tumor killing effect, and at the same time reduce the damage to normal cells of the body. 3. The high-entropy subnanowire complex of the present invention can significantly enhance the activity of oxidase-like enzymes (OXD), peroxidase (POD), and catalase (CAT) in tumor sites under ultrasonic stimulation, generating a large amount of toxic ROS to kill tumor cells. At the same time, it can also effectively remove the high concentration of GSH in tumor cells, further killing tumor cells. 4. The high-entropy subnanowire composite of the present invention has a simple preparation method with good reproducibility and is suitable for large-scale preparation. Attached Figure Description

[0020] Figure 1 This is a transmission electron microscope (TEM) image of the high-entropy subnanowire composite in Example 1 of the present invention; Figure 2 This is a high-resolution transmission electron microscope (HRTEM) image of the high-entropy subnanowire composite in Example 1 of the present invention; Figure 3 The X-ray powder diffraction (XRD) pattern of the high-entropy subnanowire composite in Example 1 of this invention; Figure 4 This is the Fourier transform infrared (FT-IR) spectrum of the high-entropy subnanowire composite in Example 1 of the present invention; Figure 5 The experimental results for verifying the ROS generation capability of the ultrasound-enhanced high-entropy subnanowire composite in Example 2 of this invention are as follows: Figure 5 a represents the high-entropy subnanowire complex that generates superoxide anions (O2·) during ultrasonic stimulation. - The result image, Figure 5 Figure b shows the results of hydroxyl radical (·OH) generation from the high-entropy subnanowire composite during ultrasonic stimulation. Figure 5 c is the graph showing the GSH consumption results of the high-entropy subnanowire composite. Figure 5 d is a graph showing the results of oxygen production from the high-entropy subnanowire composite. Figure 6 This is a graph showing the in vitro cytotoxicity test results of the high-entropy sub-nanowire complex in Example 3 of the present invention; Figure 7 This is a graph showing the biocompatibility evaluation effect of the high-entropy sub-nanowire composite in Example 4 of the present invention; Figure 8 This is a graph showing the metabolic results of the high-entropy subnanowire complex in 4T1 tumor-bearing mice in Example 5 of the present invention. Figure 9 This is a diagram showing the in vivo treatment results of the high-entropy subnanowire composite in Example 6 of the present invention, wherein... Figure 9 a is a graph showing the change in tumor volume over time in 4T1 tumor-bearing mice after treatment with different methods. Figure 9 Figure b shows the tumor inhibition rate of different treatments on mouse tumors. Detailed Implementation

[0021] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in this field or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0022] The embodiments of the present invention are described below to further illustrate the invention in detail. It should be noted that the following examples are merely examples within a suitable scope, and the specific process parameters, etc., are for reference only and are not intended to limit the specific values ​​in the examples below. Those skilled in the art can select parameters within a suitable range according to actual needs based on the description herein.

[0023] Example 1: Preparation of high-entropy subnanowire composites A PtFeMoCoNiRu-F high-entropy subnanowire composite (HESNW-F) was prepared by the following steps: S1. High-entropy subnanowires were obtained via a thermal solvent method. 98 mg of platinum acetylacetone, 70 mg of iron acetylacetone, 100 mg of molybdenum hexacarbonyl, 70 mg of cobalt acetylacetone, 38 mg of nickel acetylacetone, 50 mg of ruthenium acetylacetone, 393 mg of cetyltrimethylammonium bromide, and 600 mg of glucose were weighed and dissolved in 50 mL of oleylamine. The mixture was ultrasonically treated at room temperature for 4 h to obtain the first mixture. The first mixture was then heated to 210 °C in an oil bath and reacted for 6 h. After the reaction was completed, the mixture was cooled to room temperature, and the product was collected by centrifugation. The product was then washed 5-7 times with a cyclohexane / ethanol (volume ratio of 9:1) mixed solvent to obtain high-entropy subnanowires. The above-mentioned platinum acetylacetonate, CAS: 15170-57-7; The above-mentioned iron acetylacetone, CAS: 14024-18-1; The above-mentioned molybdenum hexacarbonyl, CAS: 13939-06-5; The above-mentioned cobalt acetylacetonate, CAS: 14024-48-7; The above-mentioned nickel acetylacetonate, CAS: 3264-82-2; The aforementioned ruthenium acetylacetone, CAS: 14284-93-6; High-entropy subnanowire composites were obtained by modification with S2 and aldehyde-modified polyether F127. 50 mg of aldehyde-modified polyether F127 and 10 mg of high-entropy subnanowires were dissolved in 20 mL of chloroform and treated in an ultrasonic water bath for 3 h to obtain a second mixture. The second mixture was rotary evaporated, and after the solvent evaporated, the resulting solid was dissolved in deionized water, thoroughly mixed in an ultrasonic water bath, and washed 3 times with deionized water to obtain the high-entropy subnanowire composite. The aforementioned aldehyde-modified polyether F127 has a relative molecular mass of 13,000, product number: P822479, and is manufactured by Shanghai Maclean Biochemical Technology Co., Ltd.

[0024] The performance of the obtained high-entropy subnanowire composite was tested, and the results are as follows: 1. Transmission electron microscopy (TEM) characterization: such as Figure 1 As shown, the obtained high-entropy subnanowire composite exhibits a linear distribution with a diameter of approximately 1 nm. 2. High-resolution transmission electron microscopy (HRTEM) characterization: such as Figure 2 As shown, the lattice width of the prepared high-entropy subnanowire composite is approximately 0.2 nm; 3. X-ray powder diffraction (XRD) test: The test results are shown in the figure below. Figure 3 As shown, a comparison with the standard card demonstrates that the high-entropy subnanowire composite is based on platinum. 4. Fourier Transmission Infrared Spectroscopy (FT-IR) Test: Results are as follows Figure 4 As shown, this demonstrates that aldehyde-modified polyether F127 was successfully loaded onto high-entropy subnanowires. 5. ICP test: The atomic ratio of Pt, Fe, Mo, Co, Ni and Ru in the prepared high-entropy subnanowire composite was 33.8:21.7:22.5:9.0:7.8:5.12.

[0025] Example 2: Verification of the ability of ultrasound-enhanced high-entropy subnanowire composites to generate ROS The high-entropy subnanowire composite prepared in Example 1 was used as the experimental material, and the superoxide anion (O2·) was monitored using electron spin resonance (ESR) technology. - The generation of hydroxyl radicals (·OH) is shown in the following figures. Figure 5 As shown in ab; oxygen production was monitored using a dissolved oxygen meter, and GSH consumption was monitored by ultraviolet analysis of absorbance changes at 412 nm. The results are as follows. Figure 5 cd shown.

[0026] The results showed that, compared with the absence of ultrasonic stimulation, the high-entropy subnanowire composite could generate O2· more efficiently under ultrasonic stimulation. - The high-entropy subnanowire complex generates more oxygen and consumes more GSH as the content of the high-entropy subnanowire complex increases. This phenomenon indicates that the cavitation effect of ultrasound accelerates electron transfer and transmission within the high-entropy subnanowire complex, further promoting ROS generation, reducing the antioxidant capacity of tumor cells, and achieving catalytic therapy.

[0027] Example 3: In vitro cytotoxicity test of high-entropy subnanowire complexes under different treatment methods The high-entropy subnanowire complex prepared in Example 1 was used as the experimental material. The cytotoxicity of the high-entropy subnanowire complex under different treatments was evaluated at the cellular level using the CCK-8 assay. The experiment was divided into four groups: control group, ultrasound group, high-entropy subnanowire complex group, and high-entropy subnanowire complex + ultrasound group.

[0028] 4T1 cells (purchased from Shanghai Fanxin Future Biotechnology Co., Ltd.) were cultured at 5 × 10⁶ cells per well. 3 The cells were seeded at a density in 96-well plates and cultured for 24 hours. The complete culture medium was then discarded. The control group and the sonication group were replaced with new complete culture medium, while the high-entropy subnanowire complex group and the high-entropy subnanowire complex + sonication group were replaced with complete culture medium containing 200 μg / mL high-entropy subnanowire complex. The cells were then incubated in a cell culture incubator (5% carbon dioxide, 37°C) for 8 hours.

[0029] Next, the ultrasound group and the high-entropy subnanowire complex + ultrasound group were subjected to intermittent ultrasound (1.0 MHz, 1 W / cm², 50% empty space, 5 min). After ultrasounding, they were placed in a cell culture incubator (5% carbon dioxide, 37 ℃) and cultured for 12 h. After culture, the culture medium was discarded and the cells were washed twice with PBS.

[0030] In each of the four experiments, 100 μL of CCK-8 reagent (10% CCK-8 diluted with serum-free medium) was added to each well. The 96-well plates were then wrapped with aluminum foil and incubated in a cell culture incubator (5% carbon dioxide, 37°C) for 15 min. The absorbance at 450 nm was recorded using a microplate reader to assess cell viability.

[0031] The results are as follows Figure 6 As shown, the high-entropy subnanowire complex exhibits different killing effects on tumor cells under different treatment methods. The high-entropy subnanowire complex group shows a killing effect on cells, while the high-entropy subnanowire complex + ultrasound group further enhances the cell killing effect. This indicates that in the tumor microenvironment, the high-entropy subnanowire complex can react with excess H2O2 in the microenvironment, and this effect is further enhanced under the action of ultrasound, significantly improving its ability to kill tumor cells.

[0032] Example 4: Biocompatibility Evaluation of High-Entropy Subnanowire Composites 1. Reagent preparation High-entropy subnanowire complex solutions: The high-entropy subnanowire complex was dissolved in PBS buffer to prepare solutions with concentrations of 50, 100, 200, and 400 μg / mL, respectively. Diluting red blood cell suspension: Add heparin sodium to mouse venous blood for anticoagulation to obtain fresh anticoagulated blood; dilute the fresh anticoagulated blood with PBS at a volume ratio of 1:10 and mix well to obtain the solution. PBS buffer (negative control); Ultrapure water (positive control).

[0033] 2. Experimental group setup Negative control group: 1 mL PBS + 0.2 mL diluted red blood cell suspension; Positive control group: 1 mL ultrapure water + 0.2 mL diluted red blood cell suspension; Experimental group: 1 mL of high-entropy subnanowire complex solution of different concentrations (50 / 100 / 200 / 400 μg / mL) + 0.2 mL of diluted red blood cell suspension.

[0034] 3. Incubation and centrifugation Place all centrifuge tubes in a constant temperature of 37°C for 1 hour, then centrifuge at 1000-1500 rpm for 10 minutes to precipitate the unlysaturated red blood cells.

[0035] 4. Hemolysis rate detection The supernatant after centrifugation was collected separately, and the absorbance (OD value) was measured at a wavelength of 545 nm using an ELISA reader. The hemolysis rate was then calculated. The formula for calculating the hemolysis rate is:

[0036] 5. Experimental Results The results are as follows Figure 7 As shown in the bar chart of color and hemolysis rate of the supernatant in the centrifuge tube, it can be seen that the hemolysis rate of the high-entropy subnanowire complex is less than 5% in the concentration range of 50-400 μg / mL; this indicates that the high-entropy subnanowire complex has almost no hemolytic effect at the tested concentration and exhibits good biocompatibility.

[0037] Example 5: In vivo metabolic experiment of high-entropy subnanowire complex Cy5.5 fluorescent molecules were electrostatically adsorbed onto a high-entropy subnanowire complex. After stirring under photoprotection for 24 h, the high-entropy subnanowire complex was washed multiple times with deionized water to remove unbound Cy5.5. Subsequently, the high-entropy subnanowire complex loaded with Cy5.5 fluorescent molecules (10 mg / kg) was injected intratumorally into a BALB / c 4T1 mammary cancer orthotopic tumor model (mice purchased from Shanghai Paijielu Technology Co., Ltd.). Fluorescence imaging was performed using an in vivo imaging system (IVIS) under oxygen anesthesia with 2% isoflurane at 6, 12, 24, and 48 h.

[0038] The results are as follows Figure 8As shown, after injection of Cy5.5-labeled HESNW-F, a significant fluorescence signal was observed in the 4T1 tumor region by IVIS, reaching a peak at 6 h. After 48 h, the major organs (heart, liver, spleen, lungs and kidneys) and the tumor were removed for ex vivo imaging, and a weak fluorescence signal was found only in the tumor region, indicating that HESNW-F undergoes normal physiological metabolism in vivo.

[0039] Example 6: Evaluation of in vivo therapeutic effect When the tumor volume in the BALB / C mouse 4T1 mammary cancer orthotopic tumor model was 50 mm 3 At approximately 10:00 AM, 4T1 tumor-bearing mice were randomly divided into four groups: a control group, an ultrasound group, a high-entropy subnanowire complex group, and a high-entropy subnanowire complex + ultrasound group; each group consisted of five mice. Treatment was administered three times consecutively, with a one-day interval between each treatment. The high-entropy subnanowire complex (prepared according to Example 1) was injected intratumorally at a dose of 10 mg / kg using PBS as the solvent. The ultrasound treatment power was 1.0 MHz, 1 W / cm², 50% vacancy, and the duration was 5 minutes.

[0040] Starting from the day after the first treatment, the size of the tumor was measured using electronic calipers and recorded every two days. Then, on the 14th day after the first treatment, the mouse tumors were removed, and the tumor volume and weight were recorded to calculate the tumor inhibition rate. Tumor inhibition rate (%) = [(average tumor weight of control group - average tumor weight of ultrasound group / high-entropy subnanowire complex group / high-entropy subnanowire complex + ultrasound group) / average tumor weight of control group] × 100%.

[0041] Statistical results are as follows Figure 9 As shown in Figure 9a, the tumor volume of 4T1 tumor-bearing mice after different treatments changes over time. Figure 9b shows the tumor inhibition rate under different treatments. The tumor inhibition rate of the high-entropy subnanowire complex group was 38.9%, while the tumor inhibition rate of the high-entropy subnanowire complex + ultrasound group was 86.0%. This indicates that the high-entropy subnanowire complex can inhibit the growth of in situ breast cancer tumors in mice and has a therapeutic effect on tumors. Furthermore, the tumor inhibition effect is significantly enhanced when the high-entropy subnanowire complex is combined with ultrasound.

[0042] The specific embodiments described herein are merely illustrative and not intended to limit the scope of the application. Those skilled in the art can make modifications to these embodiments without contributing any inventive step after reading this specification, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A high-entropy subnanowire composite, characterized in that, This includes PtFeMoCoNiRu high-entropy subnanowires and aldehyde-modified polyether F127 on the surface of PtFeMoCoNiRu high-entropy subnanowires.

2. The high-entropy subnanowire composite as described in claim 1, characterized in that, The high-entropy subnanowire composite has a width ≤1nm and a lattice width of 0.2nm.

3. The high-entropy subnanowire composite as described in claim 1, characterized in that, The atomic ratio of Pt, Fe, Mo, Co, Ni and Ru in the high-entropy subnanowire composite is 33.8:21.7:22.5:9.0:7.8:5.

12.

4. The high-entropy subnanowire composite as described in claim 1, characterized in that, The mass ratio of the aldehyde-modified polyether F127 to the high-entropy subnanowires is (4-5):

1.

5. A method for preparing a high-entropy subnanowire composite, characterized in that, Includes the following steps: S1: Platinum acetylacetonate, iron acetylacetonate, molybdenum hexacarbonyl, cobalt acetylacetonate, nickel acetylacetonate, and ruthenium acetylacetonate were added to oleylamine and mixed at room temperature to obtain a first mixture; the first mixture was heated to react, cooled, centrifuged, and then washed with a mixed solvent of cyclohexane and ethanol to obtain high-entropy subnanowires; S2: Mix high-entropy subnanowires with aldehyde-modified polyether F127 evenly to obtain a second mixture; evaporate, dissolve, and wash the second mixture to obtain the final product.

6. The method for preparing a high-entropy subnanowire composite as described in claim 5, characterized in that, In step S1, the molar ratio of platinum acetylacetone, iron acetylacetone, molybdenum hexacarbonyl, cobalt acetylacetone, nickel acetylacetone, and ruthenium acetylacetone is 0.25:0.20:0.38:0.27:0.15:0.

13.

7. The method for preparing a high-entropy subnanowire composite as described in claim 5, characterized in that, In step S2, the mass ratio of the aldehyde-modified polyether F127 to the high-entropy subnanowire is (4-5):

1.

8. The use of a high-entropy subnanowire composite as described in any one of claims 1-4 in the preparation of a medicament for tumor catalytic therapy.

9. The application as described in claim 8, characterized in that, The tumor catalytic therapy is combined with ultrasound therapy.

10. The application as described in claim 8, characterized in that, The drug is administered via intratumoral injection.

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