Black phosphorus activated paclitaxel prodrug hydrogel drug delivery system and its preparation and application

Abstract

The present invention relates to the technical field of medical nanomaterials, and provides a hydrogel drug delivery system for paclitaxel prodrug activated by black phosphorus, comprising a sodium alginate hydrogel and black phosphorus nanosheets and paclitaxel dimers loaded on the sodium alginate hydrogel. The present invention also provides a method for preparing the above-mentioned hydrogel drug delivery system and its application in the preparation of anti-tumor drugs. The present invention uses black phosphorus nanosheets as a photothermal converter and a carrier of chemotherapeutic drugs, generates ROS under near-infrared light irradiation, activates paclitaxel prodrug, synergistically fights tumors from two pathways of chemotherapy and photothermal therapy, and reduces the toxic side effects of traditional chemotherapeutic drugs, which is expected to provide a safer and more efficient treatment method for tumor patients.

Description

Technical Field

[0001] The present invention relates to the technical field of medical nanomaterials, and in particular to a hydrogel drug delivery system for paclitaxel prodrug activated by black phosphorus, and the preparation and application thereof. Background Art

[0002] Liver cancer, represented by hepatocellular carcinoma (HCC), is the fifth most common cancer in the world and the second leading cause of cancer death. Chemotherapy is the first-line treatment for advanced liver cancer, but its toxic side effects greatly limit the dosage and often lead to tumor recurrence. The tumor-selective prodrug strategy can specifically activate and release drugs in the tumor targeting area, and has outstanding advantages in improving the physicochemical, biological and pharmacokinetic properties of drugs and reducing the toxic side effects of chemotherapy drugs. Paclitaxel is a widely used chemotherapy drug in clinical practice. Its serious toxic side effects affect the quality of life of patients and even cause patients to terminate chemotherapy due to intolerance. Studies have shown that dimerized paclitaxel prodrugs can significantly reduce its toxicity, release paclitaxel under the activation of reactive oxygen species (ROS), and restore its potent anti-tumor effect. Therefore, paclitaxel dimers have become very promising prodrugs to reduce the side effects of paclitaxel.

[0003] Photothermal therapy is a cancer therapy with few side effects and high efficiency that has been developed in recent years. It mainly uses photothermal converters to convert light energy into heat energy to kill tumor cells under the irradiation of external light sources such as near-infrared lasers. It is a new non-invasive tumor treatment method. Photothermal therapy has the advantages of few adverse reactions, high specificity, and repeatable treatment. Compared with traditional tumor treatment methods, it has shown significant superiority. Black phosphorus has a wide range of light absorption and excellent photothermal conversion efficiency. In particular, it can effectively absorb near-infrared light near 800nm ​​and produce photothermal effect. Therefore, it can be used as a good photothermal material to kill tumors; black phosphorus can effectively produce ROS under 660nm near-infrared light, and then kill tumors through photodynamic therapy. In addition, due to its large specific surface area, two-dimensional black phosphorus can also be used as a carrier material to deliver chemotherapy drugs or gene drugs to the tumor site to kill tumor cells.

[0004] Therefore, we can try to combine prodrugs and photothermal therapy technology to synergistically fight tumors from both chemotherapy and photothermal therapy. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a black phosphorus activated paclitaxel prodrug hydrogel drug delivery system and its preparation and application. The system uses black phosphorus nanosheets as a photothermal converter and a carrier of chemotherapy drugs, generates ROS under near-infrared light irradiation, activates paclitaxel prodrug, synergistically fights tumors through chemotherapy and photothermal therapy, and reduces the toxic side effects of traditional chemotherapy drugs, which is expected to provide a safer and more efficient treatment method for tumor patients.

[0006] The present invention adopts the following technical solutions to solve the above technical problems:

[0007] A black phosphorus activated paclitaxel prodrug hydrogel drug delivery system (PTX2-BP@hydrogel), comprising sodium alginate hydrogel and black phosphorus nanosheets (BP) and paclitaxel dimer (PTX2) loaded on the sodium alginate hydrogel.

[0008] As one of the preferred embodiments of the present invention, the size of the black phosphorus nanosheet (BP) is 200-500 nm. The black phosphorus nanosheet has a small size and a large specific surface area. On the one hand, it has a high photothermal conversion efficiency and produces ROS effect, and on the other hand, it can also load chemotherapeutic drugs.

[0009] As one of the preferred embodiments of the present invention, the paclitaxel dimer (PTX2) is a paclitaxel prodrug reported in literature, which has a thioether bond and can be reduced under reactive oxygen species (ROS) to release paclitaxel monomers.

[0010] As one of the preferred embodiments of the present invention, the sodium alginate hydrogel is prepared from 1% sodium alginate and 1% calcium alginate in a volume ratio of 1:1. The sodium alginate hydrogel can effectively load black phosphorus and paclitaxel prodrugs, so that they can more effectively aggregate near tumor cells to exert anti-tumor effects.

[0011] As one of the preferred embodiments of the present invention, in the hydrogel drug delivery system, the mass ratio of sodium alginate, calcium alginate, black phosphorus nanosheets and paclitaxel prodrug is (1000-2000): (1000-2000): (200-600): 1.

[0012] A method for preparing the above-mentioned black phosphorus activated paclitaxel prodrug hydrogel drug delivery system, the steps are as follows:

[0013] (1) preparing black phosphorus nanosheets: dispersing black phosphorus blocks in N-methylpyrrolidone (NMP); after mixing the black phosphorus blocks, transferring them to a water bath and performing probe ultrasound for 6 to 18 hours at a water bath temperature of 5 to 20° C.; after ultrasound, centrifuging at 5000 to 10000 rpm for 5 to 30 minutes to obtain a dispersion of black phosphorus nanosheets;

[0014] (2) Preparation of sodium alginate hydrogel: 1% sodium alginate aqueous solution and 1% calcium alginate aqueous solution were mixed in a volume ratio of 1:1 and allowed to stand for 5 to 10 minutes to obtain the desired hydrogel;

[0015] (3) Preparation of black phosphorus-paclitaxel prodrug suspension: centrifuge the black phosphorus NMP suspension at 10,000-14,000 rpm for 8-12 min, remove the supernatant NMP solvent; resuspend the dispersible tablets with ddH2O, centrifuge again at 10,000-14,000 rpm for 8-12 min, repeat twice to fully remove the NMP in the suspension; finally, resuspend the black phosphorus dispersible tablets with ddH2O for standby use; mix the paclitaxel prodrug and the black phosphorus dispersible tablets at a mass ratio of 1:(200-600), and stir evenly by ultrasonication;

[0016] (4) adding the black phosphorus-paclitaxel prodrug suspension prepared in step (3) to the hydrogel in step (2), stirring and mixing, so that the suspension is evenly dispersed in the hydrogel, thereby obtaining the desired hydrogel drug delivery system.

[0017] Application of the above-mentioned black phosphorus activated paclitaxel prodrug hydrogel drug delivery system in the preparation of anti-tumor drugs.

[0018] Design principle:

[0019] Paclitaxel is a first-line treatment for patients with malignant tumors, but its toxic side effects greatly limit the dosage, often leading to tumor recurrence. The tumor-selective prodrug strategy can specifically activate and release drugs in the tumor targeting area, and has outstanding advantages in reducing the toxic and side effects of chemotherapy drugs. Black phosphorus nanomaterials are good photothermal conversion agents. Under near-infrared light irradiation, they can not only produce photothermal effects to inhibit tumors, but also release ROS to activate paclitaxel dimers, thereby releasing paclitaxel monomers to exert a strong synergistic anti-tumor effect. The present invention aims to address the problem of large toxic and side effects of paclitaxel drugs, and provide a hydrogel drug delivery system in which paclitaxel prodrugs are adsorbed on black phosphorus nanosheets and its use in the preparation of anti-tumor drugs. When the nano drug delivery system of the present invention is used to prepare anti-tumor drugs, it can achieve the aggregation of black phosphorus nanosheets loaded with paclitaxel prodrugs in tumor tissues under the carrier effect of hydrogels, and under near-infrared light irradiation, black phosphorus produces ROS effects to activate paclitaxel prodrugs to release active paclitaxel monomers, so as to achieve the photothermal effect of black phosphorus to cooperate with chemotherapy drugs to efficiently kill tumor cells and reduce damage to normal tissues (such as Figure 1 shown).

[0020] The advantages of the present invention over the prior art are as follows: the present invention adopts low-toxic paclitaxel prodrug and black phosphorus nanosheets to be co-loaded on sodium alginate hydrogel with good biosafety, and has more beneficial effects: firstly, the chemotherapeutic drug with high toxicity is designed into a prodrug (paclitaxel prodrug), which can effectively reduce its toxicity, thereby reducing the toxicity of the drug to normal cells in vivo; secondly, the paclitaxel prodrug can be activated under the excitation of black phosphorus near-infrared light, release active paclitaxel monomers, and restore its strong anti-tumor effect; moreover, black phosphorus can not only serve as an exciter of paclitaxel prodrug, but also produce a strong photothermal effect to kill tumors; finally, the present invention selects sodium alginate hydrogel as a carrier, which is safe and low-toxic, can effectively slow down the degradation of black phosphorus, and when intratumorally administered, the paclitaxel prodrug and black phosphorus can be effectively enriched near the tumor tissue to exert an anti-tumor effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is the design principle diagram of the present invention;

[0022] Figure 2 1. It is a graph showing the characterization and stability test results of the black phosphorus nanosheets of the present invention (in the graph, graph A is a scanning electron microscope image of the black phosphorus nanosheets; graph B is a transmission electron microscope image of the black phosphorus nanosheets; graph C is an ultraviolet absorption image of the black phosphorus nanosheets; and graph D is a 7-day degradation image of the black phosphorus nanosheets);

[0023] Figure 3 It is a time trend diagram of oxidation degradation of paclitaxel prodrug of the present invention (in the figure, Figure A is a diagram of oxidation-reduction mechanism of paclitaxel prodrug under hydrogen peroxide conditions; Figure B is a HPLC diagram of oxidation activation of paclitaxel prodrug);

[0024] Figure 4 The black phosphorus hydrogel of the present invention, the paclitaxel prodrug loaded on the black phosphorus hydrogel, the paclitaxel prodrug and the toxicity of paclitaxel to liver cancer cells under different conditions (in the figure, Figure A shows the toxicity of paclitaxel and paclitaxel prodrug in Huh7 cells; Figure B shows the toxicity of paclitaxel and paclitaxel prodrug in HepG2 cells; Figure C shows the toxicity of different concentrations of black phosphorus on Huh7 cells with or without 808nm irradiation; Figure D shows the toxicity of different paclitaxel prodrugs and paclitaxel prodrugs loaded on black phosphorus hydrogel on Huh7 cells; Figure E shows the toxicity of different concentrations of black phosphorus on HepG2 cells with or without 808nm irradiation; Figure F shows the toxicity of different paclitaxel prodrugs and paclitaxel prodrugs loaded on black phosphorus hydrogel on HepG2 cells);

[0025] Figure 5 It is a diagram of the ROS effect generated by the paclitaxel prodrug loaded on the black phosphorus hydrogel in liver cancer cells of the present invention (in the figure, Figure A is a flow chart of the ROS generated by the paclitaxel prodrug loaded on the black phosphorus hydrogel in liver cancer cells; Figure B is a statistical chart of the results of Figure A);

[0026] Figure 6 This is a graph showing the effect of paclitaxel prodrug loaded in black phosphorus hydrogel and other control drugs on inducing apoptosis of liver cancer cells;

[0027] Figure 7 It is a photothermal effect diagram of the paclitaxel prodrug loaded on the black phosphorus hydrogel in vivo (in the figure, Figure A is the photothermal effect produced by different drugs under 808nm light at different times; Figure B is a real picture of the photothermal effect produced by mice under different drug treatments);

[0028] Figure 8 This is a diagram showing the effect of the paclitaxel prodrug of the present invention loaded on black phosphorus hydrogel in inhibiting tumor growth in a mouse liver cancer model (in the figure, Figure A is a real picture; Figure B is a statistical chart of the results);

[0029] Fig. 9 It is the HE image of the damage to the mouse internal organs after the mouse liver cancer model was treated with different drugs of the present invention. DETAILED DESCRIPTION

[0030] The following is a detailed description of an embodiment of the present invention. This embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation method and a specific operation process are given, but the protection scope of the present invention is not limited to the following embodiment.

[0031] The black phosphorus activated paclitaxel prodrug hydrogel drug delivery system of the present invention comprises raw materials of paclitaxel prodrug (paclitaxel dimer, PTX2), black phosphorus nanosheets (BP, size 200-500nm), sodium alginate and calcium alginate.

[0032] In terms of mass ratio, the mass ratio of sodium alginate, calcium alginate, black phosphorus nanosheets and paclitaxel prodrug in the hydrogel drug delivery system is (1000-2000): (1000-2000): (200-600): 1.

[0033] Example 1

[0034] A preparation method of a black phosphorus activated paclitaxel prodrug hydrogel drug delivery system PTX2-BP@hydrogel in this embodiment:

[0035] (1) Preparation of black phosphorus nanosheets: black phosphorus blocks are dispersed in NMP; after the black phosphorus blocks are mixed, they are transferred to a water bath for probe ultrasound for 6 hours at a water bath temperature of 20° C. After ultrasound, the mixture is centrifuged at 5000 rpm for 30 minutes to obtain a dispersion of black phosphorus nanosheets.

[0036] (2) Preparation of sodium alginate hydrogel: 1% sodium alginate aqueous solution and 1% calcium alginate aqueous solution were mixed in a volume ratio of 1:1 and allowed to stand for 5 minutes to obtain the desired hydrogel.

[0037] (3) Preparation of black phosphorus-paclitaxel prodrug suspension: The black phosphorus NMP suspension was centrifuged at 10,000 rpm for 12 min to remove the supernatant NMP solvent; the dispersible tablets were resuspended in ddH2O and centrifuged again at 10,000 rpm for 12 min, and this process was repeated twice to fully remove the NMP in the suspension; finally, the black phosphorus dispersible tablets were resuspended in ddH2O for later use; the paclitaxel prodrug and the black phosphorus dispersible tablets were mixed at a mass ratio of 1:200 and ultrasonically stirred to obtain a uniform mixture.

[0038] (4) Add the black phosphorus-paclitaxel prodrug suspension prepared in step (3) to the hydrogel in step (2), stir and mix, and evenly disperse it in the hydrogel to obtain the desired hydrogel drug delivery system. Finally, in this embodiment, the mass ratio of sodium alginate, calcium alginate, black phosphorus nanosheets, and paclitaxel prodrug is 1000:1000:200:1.

[0039] Example 2

[0040] A preparation method of a black phosphorus activated paclitaxel prodrug hydrogel drug delivery system PTX2-BP@hydrogel in this embodiment:

[0041] (1) Preparation of black phosphorus nanosheets: black phosphorus blocks are dispersed in NMP; after the black phosphorus blocks are mixed, they are transferred to a water bath and probe ultrasonicated for 12 h at a water bath temperature of 15° C. After ultrasonication, the mixture is centrifuged at 8000 rpm for 20 min to obtain a dispersion of black phosphorus nanosheets.

[0042] (2) Preparation of sodium alginate hydrogel: 1% sodium alginate aqueous solution and 1% calcium alginate aqueous solution were mixed in a volume ratio of 1:1 and allowed to stand for 8 minutes to obtain the desired hydrogel.

[0043] (3) Preparation of black phosphorus-paclitaxel prodrug suspension: The black phosphorus NMP suspension was centrifuged at 12000 rpm for 10 min to remove the supernatant NMP solvent; the dispersible tablets were resuspended in ddH2O and centrifuged again at 12000 rpm for 10 min, and this process was repeated twice to fully remove the NMP in the suspension; finally, the black phosphorus dispersible tablets were resuspended in ddH2O for later use; the paclitaxel prodrug and the black phosphorus dispersible tablets were mixed at a mass ratio of 1:400 and ultrasonically stirred to obtain a uniform mixture.

[0044] (4) Add the black phosphorus-paclitaxel prodrug suspension prepared in step (3) to the hydrogel in step (2), stir and mix, and evenly disperse it in the hydrogel to obtain the desired hydrogel drug delivery system. Finally, in this embodiment, the mass ratio of sodium alginate, calcium alginate, black phosphorus nanosheets, and paclitaxel prodrug is 1500:1500:400:1.

[0045] Example 3

[0046] A preparation method of a black phosphorus activated paclitaxel prodrug hydrogel drug delivery system PTX2-BP@hydrogel in this embodiment:

[0047] (1) Preparation of black phosphorus nanosheets: black phosphorus blocks are dispersed in NMP; after the black phosphorus blocks are mixed, they are transferred to a water bath and probe ultrasonicated for 18 h at a water bath temperature of 5° C. After ultrasonication, the mixture is centrifuged at 10,000 rpm for 5 min to obtain a dispersion of black phosphorus nanosheets.

[0048] (2) Preparation of sodium alginate hydrogel: 1% sodium alginate aqueous solution and 1% calcium alginate aqueous solution were mixed in a volume ratio of 1:1 and allowed to stand for 10 min to obtain the desired hydrogel.

[0049] (3) Preparation of black phosphorus-paclitaxel prodrug suspension: The black phosphorus NMP suspension was centrifuged at 14,000 rpm for 8 min, and the supernatant NMP solvent was removed; the dispersible tablets were resuspended in ddH2O, and centrifuged again at 14,000 rpm for 8 min, and repeated twice to fully remove the NMP in the suspension; finally, the black phosphorus dispersible tablets were resuspended in ddH2O for standby use; the paclitaxel prodrug and the black phosphorus dispersible tablets were mixed at a mass ratio of 1:600, and ultrasonically stirred to obtain a uniform mixture.

[0050] (4) Add the black phosphorus-paclitaxel prodrug suspension prepared in step (3) to the hydrogel in step (2), stir and mix, and evenly disperse it in the hydrogel to obtain the desired hydrogel drug delivery system. Finally, in this embodiment, the mass ratio of sodium alginate, calcium alginate, black phosphorus nanosheets, and paclitaxel prodrug is 2000:2000:600:1.

[0051] The hydrogel drug delivery system PTX2-BP@hydrogel of the present invention mentioned in the following experimental examples refers to the product obtained by the method of the above-mentioned Example 2 unless otherwise specified.

[0052] Experimental Example 1

[0053] This experimental example is used to verify the stability of the black phosphorus hydrogel of the present invention.

[0054] Studies have shown that unmodified black phosphorus is easily oxidized and degraded in the air. The mechanisms include: 1) Under light, the more reactive superoxide anion (O2 - ) on the surface of black phosphorus; 2) O2 - Adsorption of surface phosphorus atoms and formation of P=O bonds; 3) Under the action of water molecules, the PP bonds on the surface of black phosphorus break and decompose. The present invention uses a hydrogel carrier to wrap black phosphorus, which may reduce its contact with oxygen in the air, thereby slowly degrading it. Figure 2 As shown, black phosphorus loaded in the hydrogel did not show obvious degradation within 7 days, but black phosphorus in the aqueous solution would degrade to a great extent.

[0055] Experimental Example 2

[0056] This experimental example is used to verify the oxidative activation effect of the paclitaxel prodrug of the present invention.

[0057] 0.5 mM paclitaxel prodrug was co-incubated with 10 mM hydrogen peroxide, and reaction samples were taken at different time points (0 min, 30 min, 4 h, 20 h). The peak time and mass spectrometry results of the raw material and the activated product were detected by LC-MS (liquid chromatography-mass spectrometry). Figure 3 As shown, the paclitaxel prodrug can be activated to generate paclitaxel monomer under the oxidative environment provided by hydrogen peroxide.

[0058] Experimental Example 3

[0059] This experimental example is used to verify the toxicity of the paclitaxel prodrug of the present invention loaded on black phosphorus hydrogel to different types of liver cancer cells.

[0060] Human liver cancer cells Huh7 or HepG2 were plated in 96-well plates at a cell density of 8×10 3 Inoculate and culture overnight in a constant temperature incubator at 37°C and 5% CO2. Incubate liver cancer cells with different concentrations of paclitaxel (PTX), paclitaxel prodrug (PTX2), black phosphorus hydrogel (BP@hydrogel), paclitaxel prodrug loaded on black phosphorus hydrogel (PTX2-BP@hydrogel), or paclitaxel prodrug loaded on black phosphorus hydrogel + 808nm laser irradiation (PTX2-BP@hydrogel+808nm) for 48 hours. After sufficient incubation, discard the cell supernatant, wash twice with PBS to remove the hydrogel material, then dissolve CCK8 in the culture medium and add it to each well of cells and continue incubating for 1 hour. The absorbance was measured at 450nm using an enzyme reader. The survival rate of untreated cells was 100%. The toxicity of different drugs to different liver cancer cells is as follows. Figure 4 shown.

[0061] Experimental Example 4

[0062] This experimental example is used to verify the ROS effect generated by the paclitaxel prodrug of the present invention loaded on the black phosphorus hydrogel under near-infrared irradiation.

[0063] The 2',7'-dichlorofluorescein diacetate (DCFDA) intracellular ROS detection kit was used to detect intracellular ROS generation. DCFH-DA is non-fluorescent, but can be rapidly oxidized by ROS into fluorescent molecules (dichlorofluorescein, DCF). Briefly, Huh7 cells were collected and incubated with DCEDA at 37°C for 20 minutes, and then the extracellular DCEDA was washed. The cell suspension was incubated with PTX2-BP@hydrogel at final concentrations of 25ppm, 50ppm, and 100ppm. After 808nm laser irradiation (1.0W cm-2) for 10 minutes, ROS can convert DCFDA into green fluorescent DCF that can be detected by FACS. Figure 5 As shown, black phosphorus can produce obvious ROS benefits.

[0064] Experimental Example 5

[0065] This experimental example is used to verify the effect of paclitaxel prodrug loaded on black phosphorus hydrogel in inducing apoptosis of liver cancer cells.

[0066] FITC Annexin V apoptosis detection kit was used to detect the induction of cancer cell apoptosis by BP-PTX2@hydrogel. Briefly, Huh7 cells were treated with 50 nM PTX2 or BP-PTX2@hydrogel (50 nM PTX2 and 50 ppm BP) and irradiated with 808 nm laser (1.0 W cm -2 ) for 10 min, and then for 24 h. After another 24 h of culture, the cells were washed and collected, stained with Annexin V-FITC and PI detection kits for about 20 min, and finally the proportion of cell apoptosis and necrosis was detected by flow cytometry. Figure 6 As shown, at low concentrations, PTX2 cannot effectively induce apoptosis of liver cancer cells, but under BP near-infrared activation, it can effectively increase the toxicity of PTX2, suggesting that the prodrug paclitaxel is activated to release PTX monomers to exert anti-tumor effects.

[0067] Experimental Example 6

[0068] This experimental example is used to verify the photothermal effect of the paclitaxel prodrug of the present invention loaded on black phosphorus hydrogel in a mouse liver cancer model.

[0069] To evaluate the in vivo photothermal effect of BP-PTX2@hydrogel, we established a nude mouse liver cancer tumor model. Huh7 cells were injected subcutaneously into Balb / c nude mice. When the tumor volume reached approximately 200 mm 3 At the time of the experiment, 15 tumor-bearing mice were randomly divided into 3 groups: (1) PBS group, (2) PTX2@hydrogel (5 mg / kg),

[0070] (3) BP-PTX2@hydrogel (BP 5mg / kg, PTX 25mg / kg). In laser therapy, different drugs were injected into the tumor and the irradiation wavelength was 808nm (1.0W cm -2 ). The infrared thermal imaging camera records the infrared thermal images and monitors the changes in tumor temperature over time. Figure 7 As shown, black phosphorus hydrogel can be effectively enriched near tumor tissue and can produce a rapid and strong photothermal effect under 808nm irradiation.

[0071] Experimental Example 7

[0072] This experimental example is used to verify the tumor inhibition effect of the paclitaxel prodrug of the present invention loaded on black phosphorus hydrogel in a mouse liver cancer model.

[0073] Balb / c nude mice were housed under the required conditions. All animal procedures were approved and managed by the local ethics committee and were performed in accordance with the legal guidelines of China on the protection of animal life. Huh7 subcutaneous xenograft tumors were used as animal models to evaluate the antitumor effect. When the tumor volume reached approximately 100 mm 3 The tumor-bearing mice were randomly divided into 5 groups (n=5): PBS (blank control), PTX2 (5 mg / kg), PTX2@hydrogel (5 mg / kg), BP-PTX2@hydrogel (BP 5 mg / kg, PTX25 mg / kg), BP-PTX2@hydrogel+808nm (BP 5 mg / kg, PTX25 mg / kg). Then, these drugs were injected into the tumor of the mice every two days. After 10 minutes of intratumoral irradiation, 808nm laser irradiation (1.0 W cm -2 The body weight and tumor volume of mice were measured every 2 days. The mice were killed on the 14th day, and the tumors were removed to evaluate the tumor inhibition. To explore the safety of the preparation, the main organs (heart, liver, spleen, lung, kidney) and tumors were collected, fixed with 4% paraformaldehyde solution, embedded in paraffin, sectioned and stained with hematoxylin-eosin (HE) to evaluate the potential toxicity of the main organs. Figure 8 As shown in the figure, the black phosphorus hydrogel loaded with paclitaxel prodrug exhibited a stronger anti-tumor effect under near-infrared light activation (BP-PTX2@hydrogel+808nm), and HE staining showed that the drug exhibited better safety in animals. Fig. 9 shown.

[0074] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A black phosphorus activated paclitaxel prodrug hydrogel drug delivery system, characterized in that: It comprises a sodium alginate hydrogel and black phosphorus nanosheets and paclitaxel dimer PTX2-TK loaded on the sodium alginate hydrogel; The structural formula of the paclitaxel dimer PTX2-TK is shown below: The preparation method of the black phosphorus activated paclitaxel prodrug hydrogel drug delivery system is as follows: (1) Preparation of black phosphorus nanosheets: dispersing black phosphorus blocks in N-methylpyrrolidone (NMP); after mixing the black phosphorus blocks, transferring them to a water bath and performing probe ultrasound for 6 to 18 hours at a water bath temperature of 5 to 20° C.; after ultrasound, centrifuging at 5000 to 10000 rpm for 5 to 30 minutes to obtain a dispersion of black phosphorus nanosheets; (2) Preparation of sodium alginate hydrogel: 1% sodium alginate aqueous solution and 1% calcium alginate aqueous solution were mixed in a volume ratio of 1:1 and allowed to stand for 5 to 10 minutes to obtain the desired hydrogel; (3) Preparation of black phosphorus-paclitaxel prodrug suspension: centrifuge the black phosphorus NMP suspension at 10,000-14,000 rpm for 8-12 min, remove the supernatant NMP solvent; resuspend the dispersible tablets with ddH2O, centrifuge again at 10,000-14,000 rpm for 8-12 min, repeat twice to fully remove the NMP in the suspension; finally, resuspend the black phosphorus dispersible tablets with ddH2O for standby use; mix the paclitaxel prodrug PTX2-TK and the black phosphorus dispersible tablets at a mass ratio of 1:(200-600), and stir evenly by ultrasonication; (4) adding the black phosphorus-paclitaxel prodrug suspension prepared in step (3) to the hydrogel in step (2), stirring and mixing, so that the suspension is evenly dispersed in the hydrogel, thereby obtaining the desired hydrogel drug delivery system.

2. The black phosphorus activated paclitaxel prodrug hydrogel drug delivery system according to claim 1, characterized in that: The size of the black phosphorus nanosheets is 200-500 nm.

3. The black phosphorus activated paclitaxel prodrug hydrogel drug delivery system according to claim 1, characterized in that: In the hydrogel drug delivery system, the mass ratio of sodium alginate, calcium alginate, black phosphorus nanosheets and paclitaxel prodrug is (1000-2000): (1000-2000): (200-600):

1.

4. Use of the hydrogel drug delivery system of black phosphorus activated paclitaxel prodrug according to any one of claims 1 to 3 in the preparation of anti-tumor drugs.

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