Biomimetic nano-delivery system, preparation method therefor, and use thereof

Abstract

Disclosed is a biomimetic nano-delivery system, comprising an inner core formed from a black phosphorus nanosheet and a drug, and a shell formed from an M1-type macrophage membrane. The biomimetic nano-delivery system has an anti-tumor effect, has a good photothermal effect under the irradiation of a near-infrared laser, and can actively recognize tumor cells and release the drug precisely and efficiently.

Description

A biomimetic nano-delivery system and its preparation method and application Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparation, and in particular relates to a bionic nano-delivery system and a preparation method and application thereof. Background Art

[0002] Due to the complexity, diversity, and heterogeneity of tumors, the most commonly used treatments are surgery, radiotherapy, and chemotherapy. However, these traditional approaches all have drawbacks, including the inability of surgery to completely eliminate malignant tumors, the severe toxic side effects of radiotherapy and chemotherapy on normal cells, and the development of multidrug resistance in tumor cells due to continuous chemotherapy. In recent years, nanoparticle-mediated energy conversion therapies have attracted widespread attention. Among them, thermochemotherapy mediated by photothermal conversion nanoparticles represents a new direction for nanomedicine cancer treatment. Compared to conventional materials, ultrathin 2D nanosheets, nearly a single atomic layer thick, possess unique physicochemical properties (such as efficient photothermal conversion and drug loading) and unique biological effects (such as good biocompatibility, endocytosis, and biodegradability) that many other materials lack. A nanoplatform developed based on this approach can simultaneously deliver heat and chemotherapeutic drugs to tumors for thermochemotherapy. However, limitations include short circulation times and poor stability of nanosheets once in the body. Most nanosheets are metabolized and cleared in immune cell-rich organs such as the liver, kidney, and spleen, resulting in less than 5% of nanoparticles successfully entering tumor cells.

[0003] Existing technologies use biomimetic red blood cell membranes to coat perfluorocarbons to alleviate the hypoxic environment at the tumor site. Nanodrugs primarily rely on the EPR effect to passively target and accumulate in tumors, achieving a synergistic effect of improving the hypoxic environment and radiotherapy for tumor treatment. Existing technologies also use cancer cell membranes to modify nanoparticles loaded with paclitaxel, but similar chemotherapy drugs are often highly toxic. Existing technologies also use cancer cell membranes to modify magnetic nanoparticles loaded with chemotherapy drugs, but doxorubicin is also highly toxic, and achieving high loading efficiency with magnetic nanoparticles is difficult.

[0004] Summary of the Invention

[0005] The present invention also provides a bionic nano-delivery system and a preparation method and application thereof. The bionic nano-delivery system significantly promotes cancer cell apoptosis, thereby inhibiting tumor growth.

[0006] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0007] The present invention provides a biomimetic nano-delivery system, which is a core-shell structure, wherein the core includes black phosphorus nanosheets and drugs, and the shell is a cell membrane. The biomimetic nano-delivery system of the present invention has a photothermal effect under near-infrared laser irradiation. The near-infrared light of the present invention is 780nm to 1000nm, preferably 800nm ​​to 850nm, and more preferably 808nm. The power of the near-infrared light of the present invention is 1 to 2.5W / cm 2 , preferably 1.5 to 2.0 W / cm 2 .

[0008] In the present invention, the major diameter of the black-scale nanosheets ranges from 80 to 200 nm, preferably from 80 to 150 nm, more preferably from 100 nm; the minor diameter ranges from 50 to 150 nm, preferably from 80 to 100 nm, more preferably from 90 nm.

[0009] In the present invention, the drug includes one or more of a small molecule compound, a photosensitizer, and an enzyme. The small molecule compound includes one or more of vorinostat, doxorubicin, and paclitaxel; the photosensitizer includes one or more of zinc phthalocyanine, hematoporphyrin monomethyl ether, and a fluorescent dye with good optical properties (Bodipy dye) of the boron dipyrrolidone class; and the enzyme includes one or both of a deubiquitinating enzyme and a protein degradation-related enzyme.

[0010] In the present invention, the cell membrane includes one or more of M1 macrophage membrane, Hela cell membrane, NCI-H1299 cell membrane, LLC cell membrane, B16-F10 cell membrane, 4T1 cell membrane, CAL cell membrane, and HCT cell membrane, preferably M1 macrophage membrane. The M1 macrophage membrane of the present invention is derived from mouse M1 macrophages.

[0011] In the present invention, the mass ratio of the cell membrane: the drug: the black-scale nanosheet is 1:5 to 12:1, preferably 1:6 to 10:1, and most preferably 1:8:1.

[0012] The present invention also provides a method for preparing the biomimetic nanodelivery system, comprising the following steps: preparing black-scale nanosheets, loading the black-scale nanosheets with drugs to obtain drug-loaded black phosphorus nanosheets; extracting cell membranes; and wrapping the drug-loaded black phosphorus nanosheets with the cell membranes. The cell membranes described herein are preferably M1 macrophage membranes. The cell membrane-modified black-scale nanosheets of the present invention possess functions similar to those of the source cells, such as specific recognition, long-term blood circulation, and immune evasion. For cancer treatment, nanoparticles camouflaged with activated M1 macrophage membranes have demonstrated excellent performance in the efficient targeted delivery of small molecule compounds, photosensitizers, or enzymes to tumors.

[0013] In the present invention, the black phosphorus nanosheets serve as the core. Due to the unique folded honeycomb layered structure of the black phosphorus nanosheets, their specific surface area is greatly increased, and they can efficiently and massively load anti-tumor drugs such as vorinostat, and are further included by the M1 macrophage membrane. This new type of bionic nanoparticle not only exhibits highly specific active targeting of tumor cells, but also has immune escape properties, which prolongs the drug's circulation in the body. The bionic nanodelivery system of the present invention uses 808nm near-infrared laser to irradiate the tumor site, which can achieve a high degree of aggregation at the tumor site to achieve specific targeting. Under the action of the superior photothermal conversion performance of the black scale nanosheets, the temperature of the local tumor tissue quickly rises to a threshold of 45-50°C and lasts for a period of time, so that the tumor cell membrane undergoes irreversible damage and protein denaturation, thereby inhibiting tumor growth and even ablating the tumor. The specific preparation process and application process of the bionic nanodelivery system are shown in Figure 1.

[0014] In the present invention, the preparation method of the black scale nanosheets includes the following steps: adding black phosphorus crystal powder to an N-methyl-2-pyrrolidone (NMP) solution supersaturated with sodium hydroxide to obtain a mixed solution; ultrasonically cleaning the mixed solution at an ultrasonic power of 150 to 250 W, an ultrasonic temperature of 10°C or less, and an ultrasonic time of 8 to 12 hours to obtain a brown black phosphorus dispersion; centrifuging the brown black phosphorus dispersion at 2500 to 3500 rpm and 4 to 10°C for 5 to 15 minutes, and collecting the supernatant; centrifuging the collected supernatant again at 15000 to 20000 rpm and 4 to 10°C for 5 to 15 minutes, and then washing the supernatant 2 to 4 times with deoxygenated ultrapure water. The resulting precipitate is resuspended in deoxygenated ultrapure water and stored in the dark at -15 to -20°C. The ultrasonic temperature of the present invention is 4 to 10°C, preferably 10°C. The centrifugation temperature of the present invention is preferably 4°C.

[0015] In the present invention, the preparation method of the drug-loaded black-scale nanosheets includes the following steps: mixing the black-scale nanosheets and the drug at a mass ratio of 1:5-10 and stirring for 0.5-2 days, centrifuging at 10,000-20,000 rpm and 2-6°C for 2-8 minutes to collect the precipitate, resuspending it with 8-12% DMSO solution to obtain black-scale nanosheets loaded with vorinostat, and storing it in the dark at 2-6°C.

[0016] In the present invention, the step of wrapping the drug-loaded black phosphorus nanosheets with a cell membrane comprises the following steps: mixing the cell membrane with the drug-loaded black phosphorus nanosheets, adjusting the total volume to 0.5-1.5 mL with deoxygenated ultrapure water, and stirring evenly. After sonication in an ice bath for 20-40 minutes, stirring continuously for 0.5-2 days, a cell membrane-wrapped biomimetic nanodelivery system is obtained. The drug of the present invention is preferably vorinostat. The cell membrane of the present invention is preferably an M1 macrophage membrane.

[0017] The present invention also provides the use of the biomimetic nano-delivery system or the biomimetic nano-delivery system produced by the method in the preparation of a product for treating and inhibiting tumor growth. The biomimetic nano-delivery system actively targets tumor cells and accumulates in cancer cells. After entering the cells, it rapidly heats the local tumor tissue under 808nm infrared laser irradiation, ablating the cancerous tissue while simultaneously precisely and efficiently releasing the drug, causing cancer cell apoptosis, thereby inhibiting tumor growth.

[0018] In the present invention, the biomimetic nano-delivery system is suitable for thermal therapy of tumors. The biomimetic nano-delivery system has broad application prospects in cancer, genetic diseases, and infectious diseases. The system utilizes biomimetic nanoparticles to integrate thermal therapy, chemotherapy, and tumor imaging, delivering drugs precisely and efficiently to inhibit cancer cell proliferation.

[0019] In the present invention, the biomimetic nano-delivery system has a strong photothermal conversion effect under near-infrared laser irradiation. -2 Under the irradiation conditions of 808nm infrared laser, the temperature of black scale nanosheets with experimental concentrations rapidly increased by 20 to 35°C within 6 minutes.

[0020] In the present invention, unless otherwise specified, all components or reagents are commercially available products well known to those skilled in the art.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] This invention has constructed a precise and efficient biomimetic nano-delivery system that integrates thermal chemotherapy and tumor imaging. This biomimetic nano-delivery system can specifically identify cancer cells and accumulate within them. Once inside the cells, it successfully escapes from lysosomes. Under the action of an 808nm infrared laser, it promotes the release of the drug from the carrier, effectively converting light energy into heat energy, specifically killing cancer cells and ablating cancerous tissue, thereby inhibiting tumor growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 is a flow chart of the preparation and application of the biomimetic nanodelivery system (A: flow chart of the preparation of the biomimetic nanodelivery system; B: flow chart of the biomimetic nanodelivery system inhibiting tumor growth; C: diagram of the application results of the biomimetic nanodelivery system).

[0024] Figure 2 shows the characterization results of the biomimetic nanodelivery system (A: transmission electron microscopy (TEM) images and particle size (DLS) analysis of BPNS, BS, and MBS; B: 7-day stability analysis of MBS in aqueous solution; C: Zeta potential analysis of BPNS, BS, and MBS; D: photothermal stability and photothermal conversion ability of the BP-based delivery system under repeated irradiation of 808 nm at different laser intensities; E: photothermal effect caused by different laser powers; F: photothermal effect caused by different formulations; G: cytotoxicity of MB components at different concentrations).

[0025] FIG3 shows the results of BP-induced loading efficiency analysis.

[0026] FIG4 is the results of cell viability analysis in Experimental Example 3 (A: cell viability results induced by different concentrations of SAHA; B: effects of different drug treatments on cell viability).

[0027] FIG5 shows the results of polarization index analysis of macrophages in Experimental Example 4 (A: results of polarization index detection of cells by western blotting; B: results of polarization index detection by real-time fluorescence quantitative PCR).

[0028] Figure 6 shows the results of cell uptake analysis by laser confocal microscopy and flow cytometry in Experimental Example 5 (A: results of efficient cell uptake of BPNS and MBP as shown by laser confocal microscopy and flow cytometry analysis; B: results of cell uptake of different cell lines as shown by laser confocal microscopy and flow cytometry analysis).

[0029] Figure 7 shows the confocal microscopy results of the dose-dependent and time-dependent uptake of LLC cells delivered by the MBS system in Experimental Example 6 (A: laser confocal microscopy analysis shows the uptake of LLC cells incubated with different concentrations of nanoparticles (0.62, 1.25, 2.5, 5, and 10 μg / mL); B: laser confocal microscopy shows the uptake of LLC cells incubated with BP-containing nanoparticles at 37°C for different times (1, 4, 8, 12, and 24 h)).

[0030] FIG8 is an in vitro evaluation of the inhibitory effect of MBS on tumor cells in Experimental Example 7 (A: Live / Dead assay results; B: FACS analysis results).

[0031] Figure 9 shows the evaluation of the tumor inhibition effect of MBS and the analysis results of changes in serum biochemical indicators in Experimental Example 8 (A: changes in tumor volume under different treatments; B: images of extracted tumors after different treatments; C: TUNEL analysis of tumor tissue; D: analysis of changes in mouse body weight; E: analysis of serum biochemical indicators). DETAILED DESCRIPTION

[0032] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0033] Example 1

[0034] This example is used to illustrate the preparation of the biomimetic nano-delivery system of the present invention.

[0035] (1) Preparation of black scale nanosheets (BPNS). Liquid-phase ultrasonic-assisted exfoliation method was used: First, 25 mL of N-methyl-2-pyrrolidone solution (NMP) was accurately measured with a measuring cylinder and poured into a beaker of appropriate size. Then, an appropriate amount of anhydrous sodium hydroxide was weighed and dissolved in the NMP solution, making sure that the sodium hydroxide was supersaturated. Accurately weigh 25 mg of black phosphorus crystal powder and add it to the above-mentioned sodium hydroxide supersaturated NMP solution. The obtained mixture was placed in an ultrasonic cleaning machine with an ultrasonic power of 300 W and an ultrasonic process temperature maintained below 10 ° C. The ultrasonic process was continued for 10 h. The brown black phosphorus dispersion obtained after 10 h was centrifuged in an ultracentrifuge with the parameters set to 9000 rpm and centrifuged at 4 ° C for 10 min to remove the unexfoliated macroscopic black phosphorus crystals, and the supernatant after centrifugation was carefully collected. The collected supernatant containing BPNS was then centrifuged again at 15,000 rpm and 4°C for 10 min to remove NMP. The supernatant was then washed three times with deoxygenated ultrapure water to remove any residual NMP. The resulting BPNS precipitate was transferred to a brown penicillin bottle and resuspended with an appropriate amount of deoxygenated ultrapure water. The solution was protected from light and stored at -20°C.

[0036] (2) Extraction of M1 macrophage membrane (Mem). The M1 macrophage membrane was obtained by repeated freezing and thawing. First, mouse M1 macrophages (prepared in Test Example 4) were collected. Washed three times with a PBS solution containing protease inhibitors. Add the Biyuntian Cell Membrane Protein Extraction Kit A reagent and 1X protease inhibitor to the cells, lyse the cells at 4°C for 30 minutes, and vortex once every 10 minutes. The cell suspension was frozen at -80°C for 2 hours and thawed at room temperature, repeatedly frozen and thawed four times, and ultrasonically treated at 4°C for 5 minutes at a power of 60W. The lysate was centrifuged at 14,000 rpm and 4°C for 30 minutes. The resulting precipitate was the cell membrane. The cell membrane was suspended in D-hanks solution and stored at -80°C.

[0037] (3) Preparation of M1 macrophage membrane-encapsulated black phosphorus nanosheets (MBS) loaded with vorinostat. M1 macrophage membrane (12.1 μl, 8.24 mg / ml), SAHA (400 μl, 2 mg / ml), and BPNS (50 μl, 2 mg / ml) were mixed at a mass ratio of 1:8:1. After sonication on ice for 30 min, the mixture was stirred at room temperature for 24 h to complete self-assembly. Excess membrane material was removed by centrifugation at 15,000 rpm and 4°C for 10 min. The precipitate was resuspended in an appropriate amount of 10% DMSO solution to obtain MBS. The mixture was stored at 4°C in the dark.

[0038] Example 2

[0039] This example is used to illustrate the preparation of BS and MB.

[0040] (1) Preparation of vorinostat-loaded black scale nanosheets (BS). The BPNS (1 mg / ml) prepared in Example 1 was mixed with the vorinostat antitumor drug (SAHA, 2.5 mg / ml) at a mass ratio of 1:8 and stirred for 24 h. The mixture was centrifuged at 15,000 rpm and 4°C for 5 min to collect the precipitate, which was resuspended in an appropriate amount of 10% DMSO solution to obtain BS. The mixture was protected from light and stored at 4°C.

[0041] (2) Black-scale nanosheets (MBs) coated with M1 macrophage membranes. The M1 macrophage membranes (8.24 mg / ml) prepared in Example 1 and BPNS (2 mg / ml) were mixed at a mass ratio of 1:1. After sonication in an ice bath for 30 min, the mixture was stirred at room temperature for 24 h to complete self-assembly. Excess membrane material was removed by centrifugation at 15,000 rpm and 4°C for 10 min. The precipitate was resuspended in an appropriate amount of 10% DMSO solution to obtain MBs. The mixture was stored at 4°C in the dark.

[0042] Test Example 1

[0043] This experimental example is used to illustrate the characterization and in vitro biocompatibility of the biomimetic nanodelivery system.

[0044] The BPNS, BS and MBS prepared in Example 1 were observed by transmission electron microscopy and subjected to particle size analysis. The MBSs prepared in Example 1 were subjected to a stability test according to conventional methods. The BPNS, BS and MBS prepared in Example 1 were subjected to Zeta potential analysis according to conventional operating methods. The samples prepared in Example 1 were irradiated with 808nm infrared lasers of different powers for different times, and the photothermal stability and photothermal conversion capacity of the bionic nanosystem under repeated irradiation of 808nm lasers of different laser intensities were measured. The photothermal properties of the BPNS, BS, MBS, SAHA and PBS prepared in Example 1 were investigated according to conventional operating methods. The effects of different concentrations of Mem, BPNS and SAHA on LLC cell viability were investigated according to conventional operating methods.

[0045] The results of the above experiments are shown in Figure 2. Figure 2A shows the morphology and particle size density of BPNS, BS, and MBS taken with a projection electron microscope; Figure 2B shows that the MBS aqueous solution has good stability within 7 days; Figure 2C shows the zeta point characteristics of BPNS, BS, and MBS; Figures 2D and 2E show that the use of 1.5W / cm 2 or 2W / cm 2 , the temperature rise after 10 minutes of laser irradiation is less than 45°C; Figure 2F shows that the temperature of the sample containing BPNSs is higher than that of the sample without BPNSs under the same laser power and time; Figure 2G shows that the cell membrane and black phosphorus rice sheets have good biocompatibility with cells.

[0046] Test Example 2

[0047] This experimental example is used to illustrate the BP-induced loading efficiency.

[0048] A 1ml volume of SAHA (0.31-10mg) was added to a 5ml BPNS (1mg / mL) aqueous solution. 4ml of water was then added, and the mixture was stirred in the dark for 24h. The mixture was then centrifuged (10min, 15000rpm, 4°C) to obtain BPNS / SAHA (BS). BS was then suspended in water. The results are shown in Figure 3, indicating that the optimal efficiency of SAHA loading with BPNS at a concentration of 1mg / mL is 2.5mg of SAHA, with an encapsulation efficiency exceeding 80%.

[0049] Test Example 3

[0050] This test example is used to illustrate the analysis of cell viability.

[0051] Groups: NC group (blank control without any drug added to the cells), NCL group (NC+L, blank control without any drug added to the cells), BP-L group (BPNS treatment), SAHA-L group (SAHA treatment), BS-L group (BS treatment), MBS group (5μg / mL MBS treatment), MBS-L (MBS+L, 5μg / mL MBS treatment). L in the group represents 2W / cm 2 The drug was irradiated with 808 nm near-infrared laser light, and the drug used was prepared in Example 1.

[0052] LLC cells were cultured at a rate of 1 × 10 5Cells were seeded at a density of 100 μg / well in confocal culture dishes and cultured for 12 hours. After treatment with different drugs, the treated cells were analyzed using the conventional CCK-8 method. The results are shown in Figure 4. Figure 4A shows that treatment of LLC cells with increasing concentrations of SAHA resulted in a progressive decrease in cell viability. Figure 4B shows that the prepared biomimetic nanodelivery system combined with 808 nm near-infrared laser treatment significantly inhibited cell growth.

[0053] Test Example 4

[0054] This test example is used to illustrate the polarization index of macrophages.

[0055] RAW264.7 cells were seeded in six-well plates at a density of 2 × 10 5 cells / well. After 6 hours, RAW264.7 cells were incubated with 50 ng / mL lipopolysaccharide (LPS) for 3 days to polarize into M1-like macrophages. Cells were assayed by western blotting (iNOS and CD80), and polarization indices (iNOS and TNF-α) were measured by real-time fluorescence quantitative PCR. The results, shown in Figure 5, demonstrate that 50 ng / mL LPS can effectively induce RAW264.7 cell polarization into M1 macrophages.

[0056] Test Example 5

[0057] This experimental example is used to illustrate the cellular uptake of black phosphorus nanosheets (MB) wrapped in the membrane of M1 macrophages and the targeting evaluation.

[0058] NIH3T3, MH-S, MLE-12, and LLC cells were administered (BPNS prepared in Example 1 and MB prepared in Example 2) for 8 hours and then incubated at 37°C under 5% CO2. The cells were washed with PBS, fixed with 4% paraformaldehyde for 15 minutes, stained with DAPI for 15 minutes, and analyzed by CLSM. To quantitatively analyze cellular uptake, flow cytometry was performed after trypsinization and collection of cells. The results are shown in Figure 6. Figure 6A shows that all of the above cells have an uptake effect on MB, with LLC cells having a higher uptake efficiency. Figure 6B shows that MB is more targeted to LLC cells than other cell types.

[0059] Test Example 6

[0060] This study was used to demonstrate the dose-dependent and time-dependent uptake of LLC cells delivered by the MBS system.

[0061] LLC cells were cultured in DMEM supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin and treated with MBS. The cells were washed with PBS, fixed with 4% paraformaldehyde for 15 minutes, stained with DAPI for 15 minutes, and analyzed by CLSM. The results are shown in Figure 7. Figure 7A shows that LLC cells reached optimal uptake of MBS at 5 μg / mL; Figure 7B shows that LLC cells treated with 5 μg / mL MBS reached optimal uptake after 12 hours.

[0062] Test Example 7

[0063] This test example is used to illustrate the in vitro evaluation of the inhibitory effect of MBS on tumor cells.

[0064] Groups: PBS group (buffer solution treatment), PBS-L group (buffer solution combined with laser treatment), BP group (BPNS treatment), SAHA-L (SAHA combined with laser treatment), BS-L (BS combined with laser treatment), MBS (5μg / mL MBS treatment), MBS-L (5μg / mL MBS combined with laser treatment). The laser treatment was 2W / cm 2 The cells were irradiated with 808 nm near-infrared laser for 10 min.

[0065] LLC cells were cultured at a rate of 1 × 10 5 The cells were seeded at a density of 1×10 / well in a confocal culture dish and cultured for 12 h. After treatment with different groups of drugs, the cells were collected and the live / dead cell staining was performed using a conventional detection kit and the apoptosis FACS analysis was performed using the Annexin V-FITC / PI cell apoptosis detection kit. In order to analyze cell apoptosis, LLC cells were seeded at a density of 1×10 5 The cells were plated at a density of 1 / well in a 6-well plate. The results are shown in Figure 8 . Figure 8A shows that MBS combined with 808nm near-infrared laser can more effectively inactivate LLC cells. Figure 8B shows that the MBS-L treatment group effectively inactivated LLC cells by up to 99.3%.

[0066] Test Example 8

[0067] This experimental example is used to illustrate the evaluation of MBS's tumor inhibition effect and the analysis of changes in serum biochemical indicators.

[0068] Groups: Saline group: 200L saline; Saline-L group: 200L saline combined with laser irradiation; BP-L group: 12.5mg / kg BPNS combined with laser irradiation; SAHA-L group: 5mg / kg SAHA combined with laser irradiation; BS-L group: 12.5mg / kg BPNS carrying 5mg / kg SAHA combined with BS combined with laser irradiation; MBS group: 25mg / kg Mem wrapped 12.5mg / kg BPNS carrying 5mg / kg SAHA combined with MBS; MBS-L group: 25mg / kg Mem wrapped 12.5mg / kg BP nanosheets carrying 5mg / kg SAHA combined with MBS combined with laser irradiation. Among them, L in the group represents 2W / cm 2 808nm near-infrared laser irradiation; BS and MBS in the group were prepared according to the above method.

[0069] Based on the above grouping, tumor-bearing mice were intravenously injected with different dosage forms for 12 hours, and the laser irradiation was performed at 2W / cm 2 Irradiation with 808nm near-infrared laser was performed for 10 minutes, with the light source 10cm away from the tumor site, and treatment was performed every 3 days. After 20 days, the changes in tumor volume were detected according to conventional methods, and the tumors of each treatment group were photographed. The tumor tissues of each treatment group were subjected to UNEL analysis according to conventional operating methods. The body weight of mice in each treatment group was detected according to conventional methods. The ALT content in the serum of each treatment group was detected using a mouse serum alanine aminotransferase (ALT) ELISA kit. The results are shown in Figure 9, indicating that the tumor in the MBS-L group was more significantly reduced, and the changes in body weight and serum biochemical indicators were no different from those in the other treatment groups. This shows that MBS-L treatment has better anti-tumor efficacy and safety.

[0070] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A biomimetic nano delivery system, characterized in that: The biomimetic nano delivery system is a core-shell structure, wherein the core comprises black phosphorus nanosheets and drugs, and the shell is a cell membrane.

2. The biomimetic nano delivery system according to claim 1, characterized in that The long diameter of the black-scale nanosheets ranges from 80 to 200 nm, and the short diameter ranges from 50 to 150 nm.

3. The biomimetic nano delivery system according to claim 1, characterized in that The drug includes one or more of a small molecule compound, a photosensitizer, and an enzyme.

4. The biomimetic nano delivery system according to claim 1, characterized in that The cell membrane includes one or more of an M1 macrophage membrane, a Hela cell membrane, an NCI-H1299 cell membrane, an LLC cell membrane, a B16-F10 cell membrane, a 4T1 cell membrane, a CAL cell membrane, and an HCT cell membrane.

5. The biomimetic nano delivery system according to claim 1, characterized in that The mass ratio of the cell membrane: drug: black scale nanosheet is 1:5 to 12:1 6. The method for preparing the biomimetic nano delivery system according to any one of claims 1 to 5, characterized in that: The following steps are involved: Black scale nanosheets are prepared, and drugs are loaded on the black scale nanosheets to obtain drug-loaded black phosphorus nanosheets; cell membranes are extracted; and the drug-loaded black phosphorus nanosheets are wrapped with the cell membranes.

7. The method according to claim 6, characterized in that The preparation method of the black-scale nanosheets comprises the following steps: adding black phosphorus crystal powder to an N-methyl-2-pyrrolidone solution supersaturated with sodium hydroxide to obtain a mixed solution; ultrasonically cleaning the mixed solution to obtain a brown black phosphorus dispersion; centrifuging the brown black phosphorus dispersion for the first time to collect the supernatant; centrifuging the collected supernatant again, washing with deoxygenated ultrapure water, and resuspending the obtained precipitate with deoxygenated ultrapure water, protecting from light, and storing.

8. The method according to claim 7, characterized in that The ultrasonic cleaning has a power of 150 to 250 W, a temperature below 10° C., and a time of 8 to 12 hours.

9. The method according to claim 7, characterized in that The first centrifugation parameters are set to 2500-3500 rpm, 4-10° C., and 5-15 min.

10. The method according to claim 7, characterized in that The re-centrifugation parameters are set to 15000-20000 rpm, 4-10° C., and 5-15 min.

11. The method according to claim 6, characterized in that The preparation method of the drug-loaded black-scale nanosheets comprises the following steps: mixing the black-scale nanosheets with the drug, collecting the precipitate by centrifugation, resuspending the precipitate with a DMSO solution, protecting the precipitate from light, and storing the precipitate.

12. The method according to claim 11, characterized in that The concentration of the black scale nanosheets is 0.2-1.8 mg / ml.

13. The method according to claim 11, characterized in that The concentration of the drug is 1.5-3.5 mg / ml.

14. The method according to claim 11, characterized in that The centrifugation parameters are set to 10,000-20,000 rpm, 2-5° C., and 3-10 min.

15. The method according to claim 6, characterized in that The preparation method of the cell membrane-wrapped drug-loaded black phosphorus nanosheets comprises the following steps: mixing the cell membrane with the drug-loaded black phosphorus nanosheets, adjusting the total volume to 0.5 to 1.5 mL with deoxygenated ultrapure water, and then stirring evenly, performing ice bath ultrasonication for 20 to 40 minutes, and continuously stirring and reacting for 0.5 to 2 days to obtain a biomimetic nano delivery system.

16. Use of the biomimetic nano-delivery system according to any one of claims 1 to 5 or the biomimetic nano-delivery system prepared by the method according to any one of claims 6 to 15 in the preparation of a product for treating and inhibiting tumor growth.

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