Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles, their preparation method, and applications

Abstract

This invention discloses Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles, their preparation method, and applications, belonging to the field of nanomaterial anti-tumor drug technology. The invention first synthesizes Mxene nanomaterials, then reduces them with AgNO3 to obtain an Ag3PO4 / Mxene complex, which reacts with NH2-PEG-NH2 to obtain Ag3PO4 / Mxene-PEG. Folic acid (FA) and 3-carboxypropyltriphenylphosphine bromide (TPP) are then grafted onto these compounds sequentially to obtain Ag3PO4 / Mxene-PEG-FA-TPP. The Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles prepared by this invention exhibit good biocompatibility. Furthermore, FA can bind to the highly expressed folate receptor on the surface of cervical cancer cells, targeting the nanomaterials to tumor cells. In addition, the strong positive charge on the TPP molecule promotes the binding of the nanoparticles to the negative potential of the mitochondrial membrane, further targeting the nanoparticles to mitochondria and enhancing their tumor-killing effect.

Description

Technical Field

[0001] This invention belongs to the field of nanomaterial anti-tumor drug technology, and specifically relates to anti-tumor nanoparticles that target tumor cell mitochondria and exert anti-tumor effects through photothermal and photodynamic effects, their preparation methods, and their applications in anti-tumor treatment. Background Technology

[0002] Cancer is a leading cause of death worldwide and a significant factor affecting life expectancy. Statistics show that in 2020 alone, there were approximately 19.3 million new cancer cases and 10 million cancer deaths globally. Current cancer treatments primarily include surgery, radiation therapy, chemotherapy, and immunotherapy; however, existing treatments have limited effectiveness and are accompanied by significant toxicity and side effects, making the development of new drugs urgently needed.

[0003] Nanomaterials are a rising star in the field of biomedicine for anti-tumor treatment. Their high stability, safety, large specific surface area, and ease of modification make them a novel alternative for cancer therapy. Nanomaterials can accumulate in tumor tissue through the permeability and retention effect (EPR). Under near-infrared light irradiation, nanomaterials can generate high heat, killing tumor cells through thermal ablation. Simultaneously, the electron transitions in nanomaterials under near-infrared light irradiation produce reactive oxygen species (ROS), which can also inhibit tumor cells. Therefore, the development of novel anti-tumor drugs using nanomaterials shows great promise.

[0004] While nanomaterials offer numerous advantages in anti-tumor therapy, passive targeting alone is insufficient to achieve satisfactory results; their targeting capabilities need to be enhanced. Some nanomaterials possess only photothermal or photodynamic effects, limiting their ability to kill tumor cells. Studies have shown that combined photothermal and photodynamic therapy (PTT / PDT) offers significant therapeutic advantages over single photothermal therapy. The localized high temperature induced by PTT increases tumor blood flow and oxygenation, improving the efficacy of PDT; cytotoxic reactive oxygen species generated during PDT can enhance tumor cell sensitivity to PTT. Furthermore, the poor biocompatibility and toxicity of some nanomaterials also limit their application. Given the clinical demand for more precise and efficient nanomedicines, the research and development of nanomedicines with precise targeting, high photothermal and photodynamic effects, and high safety is a key trend in nanotherapy. Summary of the Invention

[0005] In order to solve the technical problems existing in the background art, the purpose of this invention is to provide Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles with high efficiency and low toxicity targeting cervical cancer cells, their preparation method and application.

[0006] The objective of this invention is achieved through the following means:

[0007] This invention provides a method for preparing Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles, comprising the following steps:

[0008] (1) Synthesis of Mxene nanomaterials: Ti3AlC2 was added to HF aqueous solution, stirred for 2-4 days, centrifuged to collect the precipitate, washed with water and ethanol 1-3 times respectively, centrifuged to collect the precipitate, added to tetrapropylammonium hydroxide aqueous solution, stirred for 2-4 days, and centrifuged to obtain Mxene nanomaterials;

[0009] (2) Synthesis of Ag3PO4 / Mxene nanomaterials: The Mxene nanomaterials obtained in step (1) were added to distilled water, ultrasonically dispersed evenly, AgNO3 was added, and stirred for 5 to 30 minutes. Sodium dihydrogen phosphate solution was added dropwise, and stirred for 100 to 300 minutes. The mixture was filtered, washed 1 to 3 times with water and anhydrous ethanol, and dried to obtain the Ag3PO4 / Mxene complex.

[0010] (3) Synthesis of Ag3PO4 / Mxene-PEG: The Ag3PO4 / Mxene obtained in step (2) and NH2-PEG-NH2 are dissolved in an organic solvent, stirred at 0-4℃ for 1-10h, washed with water, and centrifuged to obtain Ag3PO4 / Mxene-PEG.

[0011] (4) Synthesis of Ag3PO4 / Mxene-PEG-FA: Folic acid (FA) and DCC and NHS are dissolved in an organic solvent in a molar ratio of 1:1~2:1~2. The mixture is stirred at 30~60℃ for 1~10h. Then, Ag3PO4 / Mxene-PEG obtained in step (3) is added, followed by triethylamine. The mixture is stirred at room temperature for 5~20h, centrifuged, and washed 1~3 times with DMSO and deionized water, respectively, to obtain Ag3PO4 / Mxene-PEG-FA.

[0012] (5) Synthesis of Ag3PO4 / Mxene-PEG-FA-TPP: 3-carboxypropyltriphenylphosphine bromide (TPP) and DCC and NHS are dissolved in an organic solvent in a molar ratio of 1:1 to 2:1 to 2. The mixture is stirred at 30 to 60 °C for 1 to 10 h. Then, Ag3PO4 / Mxene-PEG-FA obtained in step (4) is added, followed by triethylamine. The mixture is stirred at room temperature for 5 to 20 h, centrifuged, and washed 1 to 3 times with methanol, DMSO and deionized water, respectively, to obtain Ag3PO4 / Mxene-PEG-FA-TPP.

[0013] Based on the above technical solution, further, in step (1), the concentration of HF in the HF aqueous solution is 10-40%, the concentration of Ti3AlC2 in the solution is 0.1-0.5 g / L, the concentration of tetrapropylammonium hydroxide is 10-40 wt%, and the mass ratio of tetrapropylammonium hydroxide to Ti3AlC2 is 5:1-1:5.

[0014] Based on the above technical solution, further, the centrifugation speed in step (1) is 10000 to 30000 rpm.

[0015] Based on the above technical solution, further, in step (1), the Mxene nanomaterials obtained are washed with water and ethanol 1 to 3 times to remove residual tetrapropylammonium hydroxide.

[0016] Based on the above technical solution, further, the concentration of MXene nanomaterials in step (2) is 0.01 to 1 g / mL, and the mass ratio of AgNO3 to MXene nanomaterials is 1:10 to 1:1.

[0017] Based on the above technical solution, further, the concentration of the sodium dihydrogen phosphate solution in step (2) is 0.01 to 0.05 g / mL, and the mass ratio of NaH2PO4 to AgNO3 is 1:5 to 1:1.

[0018] Based on the above technical solution, further, the organic solvent mentioned in step (3) includes methanol and ethanol, the concentration of Ag3PO4 / Mxene is 0.1 to 1 mg / mL, and the mass ratio of Ag3PO4 / Mxene to NH2-PEG-NH2 is 1:3 to 3:1.

[0019] Based on the above technical solution, the organic solvent mentioned in step (4) further includes methanol and ethanol, and the mass ratio of folic acid to Ag3PO4 / Mxene-PEG is 1:1 to 10:1.

[0020] Based on the above technical solution, further, in step (4), the molar ratio of triethylamine to folic acid is 20:1 to 40:1.

[0021] Based on the above technical solution, further, the organic solvent mentioned in step (5) includes methanol and ethanol, and the mass ratio of 3-carboxypropyltriphenylphosphine bromide to Ag3PO4 / Mxene-PEG-FA is 1:1 to 10:1.

[0022] Based on the above technical solution, further, in step (5), the molar ratio of triethylamine to 3-carboxypropyltriphenylphosphine bromide is 20:1 to 40:1.

[0023] The present invention also provides Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles prepared by the above preparation method.

[0024] The present invention also provides the application of the above-mentioned Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in the preparation of drugs for treating tumors.

[0025] Based on the above technical solution, the tumor further includes cervical cancer.

[0026] The advantages of this invention over the prior art are as follows:

[0027] 1. Mxene nanomaterials have good electrochemical properties and photothermal conversion performance, but they can hardly generate reactive oxygen species. In this invention, Ag3PO4 is reduced onto Mxene nanomaterials to form a heterojunction between the synthesized Ag3PO4 / Mxene nanomaterials, which can promote the separation of electrons and holes, thereby generating a large amount of ROS and high heat.

[0028] 2. The Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles prepared by this invention have good biocompatibility.

[0029] 3. The Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles prepared in this invention have good biocompatibility. At the same time, FA can bind to the folic acid receptor highly expressed on the surface of cervical cancer cells, targeting the nanomaterials to tumor cells. On this basis, the strong positive charge on the TPP molecule promotes the binding of nanoparticles to the negative potential of the mitochondrial membrane, which can further target the nanoparticles to the mitochondria and improve the killing effect of the nanoparticles on tumors. Attached Figure Description

[0030] To more clearly illustrate the embodiments of the present invention, the accompanying drawings involved in the embodiments will be briefly described below.

[0031] Figure 1 The images shown are transmission electron microscope (TEM) images (ac) and scanning electron microscope (SEM) images (d) of the Ag3PO4 / Mxene nanoparticles in Example 1.

[0032] Figure 2 Infrared spectrum of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles prepared in Example 1;

[0033] Figure 3 The image shows the photothermal effect of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles after near-infrared irradiation in Example 2. In the image, a: Ag3PO4 / Mxene nanoparticles with different concentrations, b: near-infrared light experiments with different excitation powers, c: different types of nanoparticles, and d: photothermal stability of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles.

[0034] Figure 4 This is a graph showing the reactive oxygen species generation analysis of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 2;

[0035] Figure 5 The image shows the in vitro biocompatibility analysis of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 3, where a: cytotoxicity assay of nanoparticles, b: hemolysis assay of erythrocytes, and c: Calcein-AM / PI staining results.

[0036] Figure 6 This is a diagram showing the in vivo safety analysis of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 3;

[0037] Figure 7 The image shows the in vitro antitumor proliferation effect of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 4. In the image, a: different types of nanomaterials, b: different concentrations of nanomaterials, and c: Calcein-AM / PI staining results.

[0038] Figure 8 This is a diagram illustrating the lateral migration of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in vitro against tumors, as shown in Example 4.

[0039] Figure 9 This is a diagram showing the in vitro apoptosis-inducing effect of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles on tumor cells in Example 4.

[0040] Figure 10 This image shows the in vivo antitumor effect of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in mice in Example 5. Detailed Implementation

[0041] The present invention will be described in detail below with reference to the embodiments. However, the implementation of the present invention is not limited thereto. Obviously, the embodiments described below are only some embodiments of the present invention. For those skilled in the art, other similar embodiments can be obtained without creative effort and all fall within the protection scope of the present invention.

[0042] Unless otherwise specified, the experimental methods used in this invention are all conventional methods, and the experimental equipment, materials, reagents, etc. used can all be obtained commercially.

[0043] Example 1

[0044] The preparation method of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles includes the following steps:

[0045] (1) Synthesis of Mxene nanomaterials

[0046] Approximately 10 g of Ti3AlC2 powder was added to 60 mL of HF aqueous solution (HF concentration was 40%), and the mixture was magnetically stirred at room temperature for 3 days. The precipitate was collected by centrifugation and washed three times with water and ethanol, respectively. Then, the precipitate was collected by centrifugation (13000 rpm), and 60 mL of TPAOH aqueous solution (tetrapropylammonium hydroxide, 25 wt%) was added. The mixture was magnetically stirred for 3 days, and the precipitate was collected by centrifugation (20000 rpm) to obtain Mxene nanomaterials. The nanomaterials were washed three times with water and ethanol to remove residual TPAOH.

[0047] (2) Synthesis of Ag3PO4 / Mxene nanomaterials:

[0048] Add 10 g of MXene nanomaterials to 50 mL of distilled water and sonicate for 15 min. After sonication, add 3.06 g of AgNO3 and stir the solution for 15 min. Then, dissolve 1.91 g of NaH2PO4 in 50 mL of distilled water and add it dropwise to the above solution. Stir at room temperature for 240 min. Wash the filtered Ag3PO4 / MXene composite with water and anhydrous ethanol, and dry it in an oven at 60 °C for 12 h.

[0049] (3) Synthesis of Ag3PO4 / Mxene-PEG

[0050] 1 mg Ag3PO4 / Mxene and 1 mg NH2-PEG-NH2 were mixed in 2 mL of methanol, stirred at 0 °C for 4 h, washed 3 times with H2O, and centrifuged at 10000 g for 5 min to obtain Ag3PO4 / Mxene-PEG, which was then resuspended in PBS and stored at 4 °C.

[0051] (4) Synthesis of Ag3PO4 / Mxene-PEG-FA

[0052] Folic acid (FA) contains a carboxyl terminus, which can be effectively linked to the amino group of PEG. We activate the carboxyl group on the FA molecule using the DCC / NHS activation system, enabling it to link with PEG. The specific process is as follows:

[0053] Approximately 10 mg of FA was reacted with DCC / NHS in methanol (2.5 mL) at a molar ratio of 1:1.2:1.2, and stirred in the dark at 50 °C for 5 h. The reaction mixture was then combined with 1 mg of the synthesized Ag3PO4 / Mxene-PEG in methanol (2.5 mL), followed by the addition of 0.1 mL of triethylamine. The mixture was stirred at room temperature for 12 h. The resulting solution was centrifuged (10000 g) for 10 min and washed three times with DMSO and deionized water, respectively, to obtain Ag3PO4 / Mxene-PEG-FA, which was resuspended in PBS and stored at 4 °C.

[0054] (5) Synthesis of Ag3PO4 / Mxene-PEG-FA-TPP

[0055] 3-Carboxypropyltriphenylphosphine bromide (TPP) contains a carboxyl terminus, which can be effectively attached to the excess amino group on FA. The specific process is as follows:

[0056] Approximately 10 mg of TPP was reacted with DCC / NHS in methanol (2.5 mL) at a molar ratio of 1:1.2:1.2, and stirred in the dark at 50 °C for 5 h. The reaction mixture was then combined with 1 mg of the synthesized Ag3PO4 / Mxene-PEG-FA in methanol (2.5 mL), followed by the addition of 0.1 mL of triethylamine. The mixture was stirred at room temperature for 12 h. The resulting solution was centrifuged (10000 g) for 10 min and washed three times each with methanol, DMSO, and deionized water to obtain Ag3PO4 / Mxene-PEG-FA-TPP, which was resuspended in PBS and stored at 4 °C.

[0057] Figure 1 a-1c are transmission electron microscopy (TEM) images of the Ag3PO4 / Mxene nanoparticles prepared in Example 1. It can be seen that the Ag3PO4 / Mxene nanoparticles exhibit a sheet-like morphology, consistent with the morphology of Mxene sheet-like two-dimensional nanomaterials. The TEM images show that the Ag3PO4 / Mxene nanomaterials are composed of two different particle structures, namely Ag3PO4 and MXene, with a particle size of approximately 2 μm, which is consistent with the particle size of nanomaterials. Figure 1Image d shows the scanning electron microscope (SEM) morphology of Ag3PO4 / Mxene-PEG-FA-TPP. Compared to the Ag3PO4 / Mxene morphology, a large number of particles, namely FA-PEG-TPP, are doped into the layered structure. This indicates that Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles were successfully prepared.

[0058] Figure 2 The infrared spectrum of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles prepared in Example 1 shows that each component has a characteristic absorption peak, confirming the successful preparation of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles.

[0059] Example 2

[0060] Photothermal effect and reactive oxygen species detection analysis of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles

[0061] (1) Photothermal effect

[0062] a. Aqueous solutions of Ag3PO4 / Mxene nanomaterials at different concentrations (0, 32, 63, 125, 250, 500, 1000 μg / mL) were placed in 48-well plates and then subjected to near-infrared light at 808 nm (1 W / cm²). 2 Irradiate for 10 minutes, and measure the temperature every 2 minutes with a temperature measuring gun to record the temperature of the solution in different wells.

[0063] b. Change the excitation power (0.25, 0.5, 0.75, 1, 1.5 W / cm²) 2 The Ag3PO4 / Mxene nanomaterials were irradiated, and the temperature was measured every 2 minutes using a temperature measuring gun.

[0064] c. 1000 μg / mL aqueous solutions of Ag3PO4 / Mxene, Ag3PO4 / Mxene-PEG, Ag3PO4 / Mxene-FA, and Ag3PO4 / Mxene-PEG-FA-TPP nanomaterials were placed in 48-well plates, respectively, and then subjected to near-infrared light at 808 nm (1 W / cm²). 2 Irradiate for 10 minutes, and measure and record the temperature of the solution in different wells every 2 minutes using a temperature measuring gun.

[0065] d. Place 1000 μg / mL Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in a 48-well plate and perform 4 heating-cooling cycles, measuring the temperature every 3 minutes using a temperature measuring gun.

[0066] Figure 3The photothermal effect of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles after near-infrared irradiation in Example 2. Figure 3 Results a, 3b, and 3c indicate that Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles exhibit excellent photothermal effects that are dose-dependent. Figure 3 The results showed that after four heating-cooling cycles, the temperature-time curves and peak shapes of the Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles did not change significantly, indicating that the nanocomposite material has good photothermal stability.

[0067] (2) Detection and analysis of reactive oxygen species

[0068] An aqueous solution of Ag3PO4 / Mxene, Ag3PO4 / Mxene-PEG, Ag3PO4 / Mxene-FA, and Ag3PO4 / Mxene-PEG-FA-TPP (1000 μg / mL) nanoparticles was placed in a 48-well plate and mixed with 5 mM DPBF. The plate was irradiated with a laser for 10 min, and the absorbance at 410 nm was measured every 2 min.

[0069] Figure 4 The reactive oxygen species (ROS) generation capacity of the Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 2 was tested, and the results showed that the nanomaterials can generate ROS.

[0070] Example 3

[0071] Biocompatibility assessment of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles

[0072] The nanomaterial Ag3PO4 / Mxene-PEG-FA-TPP was co-cultured with cervical cancer SIHA cells and human umbilical vein endothelial cells (HUVECs) for 24 h. The cytotoxicity of the nanomaterial was assessed using the MTT assay. Cell viability analysis using Calcein-AM / PI staining was performed to determine the toxicity of the nanomaterial. The safety of the nanomaterial was evaluated by co-culturing it with erythrocytes for 4 h and detecting hemolysis.

[0073] The mice were injected with the nanomaterial Ag3PO4 / Mxene-PEG-FA-TPP (30 mg / kg) via the tail vein. Four hours after injection, the mice were sacrificed on day 14. The heart, liver, spleen, lungs, and kidneys were stained and the changes in the tissue structure of the important organs were analyzed under a microscope to assess the safety of Ag3PO4 / Mxene-PEG-FA-TPP in vivo.

[0074] Figure 5Biosafety analysis of the Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 3. Figure 5 The results showed that the nanoparticles had very low or no toxicity to SIHA cervical cancer cells and HUVEC human umbilical vein endothelial cells. Figure 5 b. The results showed that the nanoparticles did not cause hemolysis of erythrocytes, indicating that the nanoparticles Ag3PO4 / Mxene-PEG-FA-TPP have high biosafety. Figure 5 Calcein-AM / PI staining results showed that the addition of nanoparticles to in vitro cultured cells did not cause cell death, which also indicates that Ag3PO4 / Mxene-PEG-FA-TPP is biosafe.

[0075] Figure 6 The results of the in vivo safety analysis of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 3 show that the vital organ tissue structures of mice injected with Ag3PO4 / Mxene-PEG-FA-TPP via the tail vein were normal, indicating that Ag3PO4 / Mxene-PEG-FA-TPP is biosafe.

[0076] Example 4

[0077] In vitro antitumor activity of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles

[0078] In vitro studies used the MTT assay, Calcein-AM / PI live / dead cell staining, scratch assay, and flow cytometry to detect the near-infrared light-triggered antitumor effects of nanomaterials.

[0079] MTT Experiment

[0080] (1) 8000 SIHA cells were seeded into each well of a 96-well plate. After 6 hours of adhesion, Mxene-Ag3PO4, Ag3PO4 / Mxene-PEG, Ag3PO4 / Mxene-PEG-FA, and Ag3PO4 / Mxene-PEG-FA-TPP (100 μg / mL) were added. PBS buffer was added to the control group. Each treatment was repeated in three wells, with an additional three blank control wells. After 12 hours, the cells were washed three times with PBS, and 100 μl of fresh complete culture medium was added. Each well was irradiated with laser for 3 minutes. After 12 hours, each well was added with 10 μl of MTT reagent and 100 μl of DMEM complete culture medium. The cells were cultured in a cell culture incubator for 4 hours until purple crystals appeared at the bottom of the wells. All supernatant was carefully aspirated, and 100 μl of dimethyl sulfoxide (DMSO) was added. The cells were then cultured in a cell culture incubator for another 15 minutes to dissolve the crystals. The OD value at 570 nm was measured using a microplate reader, and cell viability was calculated.

[0081] (2) 8000 SIHA cells were seeded in each well of a 96-well plate. After 6 hours of adhesion, 100 μL of Ag3PO4 / Mxene nanomaterial (6, 12.5, 25, 50, 100, 200 μg / mL) was added. After 12 hours, the cells were washed three times with PBS and 100 μL of fresh complete culture medium was added. Each well was irradiated with near-infrared light for 3 minutes. After 12 hours, the cells were detected by MTT assay using the same method as above.

[0082] Calcein-AM / PI live / dead cell staining

[0083] 8000 SIHA cells were seeded into each well of a 96-well plate. After 6 hours of adhesion, Mxene-Ag3PO4, Ag3PO4 / Mxene-PEG, Ag3PO4 / Mxene-PEG-FA, and Ag3PO4 / Mxene-PEG-FA-TPP (100 μg / mL) were added. The control group was treated with PBS buffer. After 12 hours, the cells were washed three times with PBS, and 100 μl of fresh complete culture medium was added. Each well was irradiated with laser for 3 minutes. After 4 hours, Calcein-AM / PI staining was performed for 15 minutes, and the cells were observed under a microscope.

[0084] Scratch test

[0085] (1) 8000 cervical cancer SIHA cells were seeded in a 96-well plate and cultured in a cell culture incubator containing 5% CO2 at 37°C for 6 hours.

[0086] (2) Add 100 μg / mL of nanomaterial to SIHA cells, add PBS buffer to the control group, and culture for 12 h;

[0087] (3) Wash with PBS 3 times, add fresh complete culture medium, irradiate with near-infrared light for 3 minutes, perform scratching operation in well plate, wash with PBS 1-2 times, and add culture medium again.

[0088] (4) Inverted microscope to take pictures and record the migration of SIHA breast cancer cells from 0 to 24 hours.

[0089] Flow cytometry

[0090] (1) SIHA cells were fed at a rate of 2 × 10 5 The concentration of the medium was spread in a 6-well plate, and after adhering to the wall for 24 hours, nanomaterials were added.

[0091] (2) After culturing for 12 hours, the culture medium was aspirated and washed three times with PBS buffer.

[0092] (3) SIHA cells were digested with 0.25% trypsin without EDTA to remove them from the cell wall. After washing the SIHA cells twice in PBS buffer, they were stained with Annexin V-FITC / PI kit.

[0093] (4) Resuspend the cells in 500 μL Annexin V binding buffer, and stain SIHA cells with 5 μL each of Annexin V-FITC and PI. After staining for 30 min in the dark at room temperature, collect and detect cells with >10 cells using a Calibur flow cytometer. 4 SIHA cells.

[0094] Figure 7 This demonstrates the antitumor effect of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 4. Figure 7 The MTT assay results in samples a and 7b showed that the composite nanomaterials loaded with PEG, FA, and TPP exhibited the strongest phototoxicity, with cell viability below 40%, and this effect was concentration-dependent; the higher the concentration, the lower the cell viability. Figure 7 The Calcein-AM / PI staining results in c also confirm this.

[0095] Figure 8 The scratch assay results for the antitumor effect of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in Example 4 show that the lateral migration ability of cells treated with Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles was significantly inhibited.

[0096] Figure 9 The results of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles inducing tumor cell apoptosis in Example 4 show that the PBS group and its light-illuminated group hardly induced cell apoptosis. In the Ag3PO4 / Mxene+ light-illuminated group, the early apoptosis rate was 0.52% and the late apoptosis rate was 12.1%; in the Ag3PO4 / Mxene-PEG+ light-illuminated group, the early apoptosis rate was 1.47% and the late apoptosis rate was 15.3%; in the Ag3PO4 / Mxene-PEG-FA+ light-illuminated group, the early apoptosis rate was 2.05% and the late apoptosis rate was 18.0%; and in the Ag3PO4 / Mxene-PEG-FA-TPP+ light-illuminated group, the early apoptosis rate was 11.4% and the late apoptosis rate was 25.1%. This indicates that the composite nanomaterial Ag3PO4 / Mxene-PEG-FA-TPP can better induce cell apoptosis.

[0097] Example 5

[0098] In vivo antitumor activity of Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles

[0099] In vivo studies were conducted by subcutaneously inoculating female Balb / c mice with SIHA cervical cancer xenografts, followed by tail vein injection of the composite nanomaterial Ag3PO4 / Mxene-PEG-FA-TPP. The xenografts were then irradiated with near-infrared light at a wavelength of 808 nm. Tumor size was analyzed, and the in vivo antitumor effect of Ag3PO4 / Mxene-PEG-FA-TPP was evaluated. The specific procedures are as follows:

[0100] After passage, SIHA cervical cancer cells were centrifuged to obtain a cell pellet. The pellet was mixed with 5% glucose solution and the cells were counted at a rate of 5 × 10⁶ cells per mouse. 7 SIHA cells were collected in 100 μL suspensions and dispensed into EP tubes. Eight-week-old female Balb / c mice were randomly divided into 5 groups (n=5). SIHA cells were extracted from the EP tubes using a sterile syringe and injected into the left back of the mice. When the tumor volume reached 80 mm², the cells were injected into the left back of the mice. 3 Mice were injected with 30 mg / kg of nanomaterial via the tail vein, while the control group was injected with 5% glucose solution. Four hours after nanomaterial injection, mice were exposed to near-infrared light at a wavelength of 808 nm (3 min, 2W cm⁻¹) -2 Balb / c mice were irradiated with irradiated dorsal tumors. The weight of each group of mice was measured daily. Mice were sacrificed on day 14 after nanomaterial treatment, and organs (heart, liver, spleen, lung, kidney) and tumors were collected. The tumors were photographed to evaluate the inhibitory effect on tumor growth.

[0101] Figure 10 The results of the Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles in vivo against xenografts in mice in Example 5 show that the Ag3PO4 / Mxene-PEG-FA-TPP composite nanomaterials significantly inhibited the proliferation of cervical cancer SIHA cells in vivo.

[0102] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles, characterized in that, Includes the following steps: (1) Synthesis of Mxene nanomaterials: Ti3AlC2 was added to HF aqueous solution, stirred for 2-4 days, centrifuged to collect the precipitate, washed, centrifuged to collect the precipitate, added to tetrapropylammonium hydroxide aqueous solution, stirred for 2-4 days, and centrifuged to obtain Mxene nanomaterials; (2) Synthesis of Ag3PO4 / Mxene nanomaterials: The Mxene nanomaterials obtained in step (1) were added to water, ultrasonically dispersed evenly, AgNO3 was added, and stirred for 5 to 30 minutes. Sodium dihydrogen phosphate solution was added dropwise, and stirred for 100 to 300 minutes. The mixture was filtered, washed, and dried to obtain the Ag3PO4 / Mxene complex. (3) Synthesis of Ag3PO4 / Mxene-PEG: The Ag3PO4 / Mxene obtained in step (2) and NH2-PEG-NH2 are dissolved in an organic solvent, stirred at 0-4℃ for 1-10h, washed with water, and centrifuged to obtain Ag3PO4 / Mxene-PEG. (4) Synthesis of Ag3PO4 / Mxene-PEG-FA: Folic acid (FA) and DCC and NHS are dissolved in an organic solvent in a molar ratio of 1:1~2:1~2. The mixture is stirred at 30~60℃ for 1~10h. Then Ag3PO4 / Mxene-PEG obtained in step (3) is added, followed by triethylamine. The mixture is stirred at room temperature for 5~20h, centrifuged, and washed to obtain Ag3PO4 / Mxene-PEG-FA. (5) Synthesis of Ag3PO4 / Mxene-PEG-FA-TPP: 3-carboxypropyltriphenylphosphine bromide (TPP) and DCC and NHS are dissolved in an organic solvent in a molar ratio of 1:1 to 2:1 to 2. The mixture is stirred at 30 to 60 °C for 1 to 10 h. Then, Ag3PO4 / Mxene-PEG-FA obtained in step (4) is added, followed by triethylamine. The mixture is stirred at room temperature for 5 to 20 h, centrifuged, and washed to obtain Ag3PO4 / Mxene-PEG-FA-TPP.

2. The preparation method according to claim 1, characterized in that, In step (1), the concentration of HF in the HF aqueous solution is 10-40%, the concentration of Ti3AlC2 in the solution is 0.1-0.5 g / L, the concentration of tetrapropylammonium hydroxide is 10-40 wt%, and the mass ratio of tetrapropylammonium hydroxide to Ti3AlC2 is 5:1-1:

5. The obtained Mxene nanomaterials are washed with water and ethanol 1-3 times to remove residual tetrapropylammonium hydroxide.

3. The preparation method according to claim 1, characterized in that, The concentration of MXene nanomaterials in step (2) is 0.01 to 1 g / mL, and the mass ratio of AgNO3 to MXene nanomaterials is 1:10 to 1:

1.

4. The preparation method according to claim 1, characterized in that, The concentration of the sodium dihydrogen phosphate solution mentioned in step (2) is 0.01 to 0.05 g / mL, and the mass ratio of NaH2PO4 to AgNO3 is 1:5 to 1:

1.

5. The preparation method according to claim 1, characterized in that, The organic solvents mentioned in step (3) include methanol and ethanol, the concentration of Ag3PO4 / Mxene is 0.1 to 1 mg / mL, and the mass ratio of Ag3PO4 / Mxene to NH2-PEG-NH2 is 1:3 to 3:

1.

6. The preparation method according to claim 1, characterized in that, The organic solvents mentioned in step (4) include methanol and ethanol, the mass ratio of folic acid to Ag3PO4 / Mxene-PEG is 1:1 to 10:1, and the molar ratio of triethylamine to folic acid is 20:1 to 40:

1.

7. The preparation method according to claim 1, characterized in that, The organic solvents mentioned in step (5) include methanol and ethanol, the mass ratio of 3-carboxypropyltriphenylphosphine bromide to Ag3PO4 / Mxene-PEG-FA is 1:1 to 10:1, and the molar ratio of triethylamine to 3-carboxypropyltriphenylphosphine bromide is 20:1 to 40:

1.

8. Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles prepared by the preparation method according to any one of claims 1-7.

9. The use of the Ag3PO4 / Mxene-PEG-FA-TPP nanoparticles according to claim 8 in the preparation of drugs for treating tumors.

10. The application according to claim 9, characterized in that, The tumors mentioned include cervical cancer.

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