Hybrid cell membrane modified bionic nanoparticles with anti-tumor effect as well as preparation method and application of hybrid cell membrane modified bionic nanoparticles

Abstract

The invention discloses a hybrid cell membrane modified bionic nanoparticle with an anti-tumor effect and a preparation method and application thereof, and belongs to the technical field of pharmaceutical preparations, the bionic nanoparticle comprises a nanoparticle core and a hybrid cell membrane coating the nanoparticle core; the nanoparticle core comprises an amphiphilic polymer drug carrier and an active drug, and the active drug comprises a photo-thermal agonist TPT and an STING agonist DMXAA; the hybrid cell membrane is formed by co-extruding and hybridizing a cancer cell membrane and a macrophage membrane. The STING agonist DMXAA and the photo-thermal agonist TPT synergistically play a role, heat can be generated under near-infrared laser irradiation to kill cancer cells and induce immunogenic death of the cancer cells, and an injury-related molecular pattern is released, so that DMXAA-mediated immunotherapy is activated, and the anti-tumor effect is achieved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical preparations, and particularly relates to a hybrid cell membrane modified biomimetic nanoparticle with an anti-tumor effect, a preparation method thereof and application thereof. BACKGROUND

[0002] Cancer is a malignant disease caused by abnormal regulation of cell growth and proliferation, and its essence is the process of normal cells gradually transforming into tumor cells through multiple steps. A decisive feature of cancer is the rapid generation of abnormal cells, which grow beyond their usual boundaries and can invade nearby parts of the body and spread to other organ parts.

[0003] Photothermal therapy has become a non-invasive treatment method in the field of local tumor treatment due to its high precision, non-invasiveness, and small side effects. It usually relies on photothermal conversion agents to generate enough heat under near-infrared light irradiation to inhibit or even ablate tumors. Combination therapy can combine the advantages of different cancer treatment methods to maximize treatment effectiveness. Tumor antigen recognition and phagocytosis are key steps in immunotherapy. If photothermal therapy and immunotherapy are combined, it is expected to significantly improve anti-tumor efficacy through a two-way synergistic mechanism of "heat therapy sensitization-immune activation".

[0004] Encapsulating anticancer drugs into nanoparticles can enhance drug accumulation in diseased areas through retention and penetration, thereby enhancing treatment effectiveness. However, nanoparticles are exogenous substances that can be easily recognized and eliminated by the immune system. Inspired by the composition and transport process of cell membranes, modifying the surface of nanoparticles with endogenous cell membranes can endow them with excellent biological interface properties and long circulation ability, and further enhance immune evasion and targeting ability.

[0005] Chinese patent document CN109078183A discloses a biomimetic multifunctional nanoparticle based on cancer cell membranes. The nanoparticle is composed of a nanoparticle drug particle core and an outer layer of cancer cell membranes. The core is formed by the chemotherapeutic drug desalted doxorubicin and the photothermal therapy drug indocyanine green, and the outer layer is formed by a shell structure of human cervical cancer cell membranes.

[0006] Chinese patent document CN119185578A discloses a lung cancer cell membrane biomimetic targeted noble metal-based PLGA nanoparticle drug-loaded particle. It includes a drug-loaded core and a lung cancer cell membrane coated on the outside of the core. The drug-loaded core structure includes a core inner layer and a core intermediate layer in sequence. The core inner layer is a chemotherapeutic drug cisplatin, and the core intermediate layer is PLGA with good biocompatibility. The outer surface of PLGA is modified with a noble metal, which is one or two of gold, ruthenium, and platinum. The nanoparticle drug-loaded particle can achieve the effect of chemotherapy-photothermal synergistic therapy.

[0007] Although the above inventions all involve the preparation of biomimetic nanoparticles, they all use single cell membranes, and the stability and targeting need to be further improved. SUMMARY

[0008] In order to solve the above-mentioned problems in the prior art, the present application provides a hybrid cell membrane modified biomimetic nanoparticle with anti-tumor effect, which has good targeting and biocompatibility, immune escape ability and good anti-tumor effect.

[0009] The specific technical solutions adopted are as follows:

[0010] A hybrid cell membrane modified biomimetic nanoparticle with anti-tumor effect, comprising a nanoparticle core and a hybrid cell membrane covering the nanoparticle core.

[0011] The nanoparticle core comprises an amphiphilic polymer drug carrier and an active drug, and the active drug comprises a photothermal agonist TPT and a STING agonist DMXAA; the mass ratio of the photothermal agonist TPT, the STING agonist DMXAA and the amphiphilic polymer drug carrier is 1:10:10-15.

[0012] The structural formula of the photothermal agonist TPT is:

[0013]

[0014] The structural formula of the STING agonist DMXAA is:

[0015]

[0016] The hybrid cell membrane is formed by co-extrusion of cancer cell membranes and macrophage membranes.

[0017] The photothermal agonist TPT can generate local high heat in tumor cells under the irradiation of near-infrared laser, thereby directly killing tumor cells, inducing immunogenic cell death (ICD), releasing damage-associated molecular patterns (DAMPs), such as exposing calreticulin (CRT), releasing high mobility group protein 1 (HMGB1), and releasing ATP, which promotes the maturation of dendritic cells, enhances tumor antigen presentation, and further activates T cell-mediated anti-tumor immunity. As a STING pathway agonist, DMXAA can be taken up by antigen-presenting cells in the tumor microenvironment, promote the secretion of type I interferons, enhance CD8 +T cell and NK cell tumor killing ability; promote M1 type macrophage polarization, and then improve the immune suppression microenvironment. The ICD induced by the photothermal agonist TPT provides tumor antigens, and the STING pathway activated by DMXAA can enhance the immune system's response to tumor antigens, achieving the enhancement of the systemic anti-tumor immunity of "antigen release + immune activation", forming a positive feedback cycle for photothermal activated immunotherapy induced by the biomimetic nanoparticles. Not only improves the local tumor control rate, but also can inhibit the distant metastasis of tumors through systemic anti-tumor immunity.

[0018] Single cell membranes usually only have a single function, and hybrid cell membranes can integrate inflammation chemotaxis, homologous targeting, in vivo long circulation and other biological functions by fusing cell membranes from different sources, and can also balance immune regulation to avoid excessive activation or inhibition of immune response, providing a more precise and efficient solution for nanomedicine delivery.

[0019] Preferably, the amphiphilic polymer drug carrier is distearoyl phosphatidylethanolamine-polyethylene glycol, and further preferably DSPE-PEG 2000 .

[0020] Preferably, the hybrid cell membrane is obtained by co-extruding 4T1 cell membrane and RAW cell membrane, the 4T1 cell membrane is the cell membrane of 4T1 mouse breast cancer cells, and the RAW cell membrane is the cell membrane of RAW264.7 mouse monocyte macrophages.

[0021] Further, the hybrid cell membrane is obtained by co-extruding 4T1 cell membrane and RAW cell membrane at a ratio of 1-3:1 of the mass of membrane proteins.

[0022] The application also provides a preparation method of the hybrid cell membrane modified biomimetic nanoparticles with anti-tumor effect, comprising the following steps:

[0023] Step S1: under the action of ice bath and ultrasonic, drop the organic solution of photothermal agonist TPT and STING agonist DMXAA into the aqueous solution of amphiphilic polymer, so that the amphiphilic polymer coats the photothermal agonist TPT and the STING agonist DMXAA, remove the organic solvent and moisture, and obtain the nanoparticle inner core DDT;

[0024] Step S2: co-extrude the cancer cell membrane and the macrophage membrane in a liposome extruder to obtain the hybrid cell membrane;

[0025] Step S3: co-extrude the nanoparticle inner core and the hybrid cell membrane to obtain the hybrid cell membrane modified biomimetic nanoparticles with anti-tumor effect.

[0026] Further, the organic solvent in the organic solution of the photothermal agonist TPT and the STING agonist DMXAA is tetrahydrofuran.

[0027] Further, the preparation method of the cancer cell membrane is as follows: collecting cell precipitate by centrifugation, then resuspending by adding a hypotonic lysis buffer, incubating in an ice bath, repeatedly freezing and thawing, collecting supernatant by low-speed centrifugation, and collecting precipitate by high-speed centrifugation of the obtained supernatant to obtain the cancer cell membrane.

[0028] The application further provides application of the hybrid cell membrane modified biomimetic nanoparticle with the antitumor effect in preparation of a photothermal activation immunotherapy preparation for tumors.

[0029] Further, the tumor is a breast tumor.

[0030] Compared with the prior art, the application has the following beneficial effects:

[0031] (1) The hybrid cell membrane gives the biomimetic nanoparticle a synergistic targeting ability. The RAW cell membrane migrates to the tumor microenvironment through receptor orientation, and the 4T1 cell membrane homologously recognizes tumor cells, realizes double-stage targeting of "chemotaxis + recognition", and can accurately deliver active drugs to tumors.

[0032] (2) The hybrid cell membrane gives the biomimetic nanoparticle an immune regulation ability. The RAW cell membrane promotes M1 type macrophage polarization and enhances antigen presentation; the CD47 on the surface of the 4T1 cell membrane gives the nanoparticle a partial escape ability, and realizes synergistic activation of innate-adaptive immunity.

[0033] (3) The hybrid cell membrane gives the biomimetic nanoparticle excellent drug delivery ability. The RAW cell membrane improves vascular penetration ability, so that the biomimetic nanoparticle penetrates into the tumor microenvironment; the 4T1 cell membrane enhances tumor cell uptake, and realizes synergistic release of the photothermal agonist TPT and the STING agonist DMXAA in space.

[0034] (4) The photothermal agonist TPT and the STING agonist DMXAA synergistically activate immunity. The photothermal agonist TPT induces ICD, releases damage-associated molecular patterns, and promotes dendritic cell maturation; and the DMXAA activates the STING pathway, promotes type I interferon secretion, and enhances the tumor killing ability of CD8 + T cells and NK cells. The responses of the two immune pathways form a closed loop of "antigen release-antigen presentation-immune response", which further enhances systemic anti-tumor immunity. BRIEF DESCRIPTION OF DRAWINGS

[0035] Figure 1Particle size distribution of DDT-Se nanoparticle core and DDT-S nanoparticle core was determined by dynamic light scattering instrument (DLS).

[0036] Figure 2 Particle size distribution of DDT-HM biomimetic nanoparticle of Example 1 was determined by dynamic light scattering instrument (DLS).

[0037] Figure 3 Transmission electron microscope images of DDT-Se nanoparticle core, DDT-S nanoparticle core and DDT-HM biomimetic nanoparticle.

[0038] Figure 4 Confocal images of mixed and co-extruded DiD-labeled RAW264.7 cell membrane and DiO-labeled 4T1 cell membrane (scale bar: 50 μm).

[0039] Figure 5 Flow cytometry histograms of L929, U87, MCF-7, RAW264.7 and 4T1 cells co-cultured with DDT-HM biomimetic nanoparticles with different membrane protein ratios for 4 h.

[0040] Figure 6 Uptake of L929, U87, MCF-7, RAW264.7 and 4T1 cells by DDT-CM, DDT-MM and DDT-HM biomimetic nanoparticles (membrane protein mass ratio of HM is 1:1).

[0041] Figure 7 Optimal fusion ratio of vesicles was determined by using CD44 and CD206 to label cancer cell membrane and macrophage membrane respectively and FACS analysis.

[0042] Figure 8 Cell survival rate histograms of 4T1 cells treated with different concentrations (0.05, 0.1, 0.15, 0.2, 0.25 μg / mL) of DDT-HM biomimetic nanoparticles with or without 808 nm laser (0.8 W / cm 2 ) irradiation for 3 min.

[0043] Figure 9 Flow cytometry detection of CRT exposure on the surface of 4T1 cells treated with different drugs.

[0044] Figure 10 Immunofluorescence images of CRT exposure on the surface of 4T1 cells treated with different materials (scale bar: 100 μm).

[0045] Figure 11 Flow cytometry detection of HMGB1 efflux on the surface of 4T1 cells treated with different materials.

[0046] Figure 12 The primary tumor volume of 4T1 tumor-bearing mice after treatment by different materials. DETAILED DESCRIPTION

[0047] In order to make the objects, technical solutions and advantages of the present application clearer, further detailed description will be made to the present application in combination with the accompanying drawings and examples. It should be understood that the specific examples described herein are only used to explain the present application, and are not used to limit the present application. In addition, the technical features involved in each embodiment of the present application described below can be combined with each other as long as they do not conflict with each other.

[0048] Photo-thermal conversion agent TPT It can be synthesized according to the description in the literature Zhou, X.; Zhang, K.; Yang, C.; Pei, Y.; Zhao, L.; Kang, X.; Li, Z.; Li, F.; Qin, Y.; Wu, L., Ultrabright and Highly Polarity-Sensitive NIR-I / NIR-II Fluorophores for the Tracking of Lipid Droplets and Staging of Fatty Liver Disease. Adv. Funct. Mater. 2021, 32, 2109929.

[0049] STING agonist DMXAA The CAS number is 117570-53-3, and it is purchased from Sinoreagent Co., Ltd.

[0050] Example 1

[0051] Step 1, preparation of DDT nanoparticle core:

[0052] 1 mg of photo-thermal conversion agent TPT-Se and 10 mg of STING agonist DMXAA were weighed and dissolved in 3 mL of tetrahydrofuran (THF) solution to obtain an organic phase solution. 15 mg of DSPE-PEG 2000 was dissolved in 10 mL of water to obtain an aqueous solution. The DSPE-PEG 2000 aqueous solution was placed in an ice bath, and the TPT and DMXAA mixed organic solution was added dropwise into the DSPE-PEG 2000 aqueous solution under ultrasonic at a power of 26 W using a 1 mL syringe to obtain a uniformly dispersed micellar dispersion.

[0053] The obtained micellar dispersion liquid was transferred into a sample bottle with a magnet, placed on a stirring table to stir for 24 h to remove THF solvent, and then freeze-dried to remove water to obtain DDT nanoparticle core (DDT-Se, the preparation method of DDT-S nanoparticle core is the same as the above steps, and TPT-Se is replaced by TPT-S).

[0054] Step 2, preparation of DDT-HM biomimetic nanoparticles:

[0055] 4T1 cells (4T1 mouse breast cancer cells) and RAW cells (RAW264.7 mouse mononuclear macrophages) were incubated in DMEM medium supplemented with 10% FBS, 1% penicillin (50.0 IU mL -1 ), streptomycin (50.0 IU mL -1 ), and gentamicin solution (50.0 IU mL -1 ) (temperature: 37°C, saturated humidity: 5% CO2). When the cells were fully grown in the culture dish, the supernatant was removed, the adherent cells were gently washed with PBS, and the cells were scraped off with a cell scraper, centrifuged to collect the cells, the supernatant was removed, and the cell pellet was left. The cell pellet was resuspended with pre-cooled PBS and centrifuged for 5 minutes to collect the pure pellet. Then resuspend with hypotonic lysis buffer (membrane protein lysis buffer and protease inhibitor PMSF). After incubation in an ice bath for 10-15 min, the 4T1 cells and RAW cells were repeatedly frozen and thawed three times in liquid nitrogen and at 37°C, respectively. The frozen and thawed mixture was centrifuged at 700g for 10 minutes at 4°C, the supernatant was collected, and then centrifuged at 14000g for 30 minutes at 4°C, the supernatant was discarded, and the final 4T1 cell membrane (4T1 CM) pellet and RAW cell membrane (RAW MM) were collected. The mass of the cell membrane was determined by BCA protein content detection kit.

[0056] Then the 4T1 CM and RAW MM were co-extruded in a liposome extruder to obtain hybrid cell membranes, and the hybrid cell membranes were mixed with the DDT nanoparticle core (the mass ratio of membrane protein of 4T1 CM, membrane protein of RAW MM, and DDT nanoparticle core was 3:1:40, DDT-Se) to coat the hybrid cell membranes on the surface of DDT by repeated mechanical extrusion through a 200 nm polycarbonate porous membrane to form hybrid cell membrane modified biomimetic nanoparticles DDT-HM with anti-tumor effect.

[0057] Example 2

[0058] (1) Characterization of particle size and morphology of DDT-HM biomimetic nanoparticles:

[0059] The DDT-HM biomimetic nanoparticle dispersion prepared in Example 1 was diluted to 1 mg / mL, and 2 mL of the dispersion was used to measure the particle size and zeta potential by dynamic light scattering (DLS). After measuring the particle size, an appropriate amount of the solution was added dropwise to a 200-mesh carbon film copper mesh, which was dried in an oven at 55°C, and then the sample was observed under a transmission electron microscope (TEM) to select the morphology of the nanoparticles.

[0060] Meanwhile, two kinds of nanoparticle cores DDT-S or DDT-Se can be prepared according to the method of Example 1 due to the difference of the photothermal stimulant TPT. The particle size and morphology of DDT-S and DDT-Se were also characterized.

[0061] As shown in Figures 1-3 : The particle size of DDT is slightly different due to the difference of the encapsulated molecules, the particle size of DDT-Se is about 87 nm, the particle size of DDT-S is about 90 nm, and the particle size of DDT-HM biomimetic nanoparticles of Example 1 is 113 nm. It is observed that all of them are uniform spherical morphology in TEM, and it is also observed that the surface of DDT-HM biomimetic nanoparticles is 2-3 nm of cell membrane.

[0062] (2) Characterization of hybrid cell membrane HM:

[0063] The extracted RAW cell membrane was specifically labeled with a far-red fluorescent probe for cell membrane (DiD), and the extracted 4T1 cell membrane was specifically labeled with a green fluorescent probe for cell membrane (DiO). After incubation for 1 h, the labeled hybrid cell membrane HM was prepared by a liposome extruder, and finally observed by a laser confocal microscope.

[0064] As shown in Figure 4 : The red fluorescence of RAW cell membrane and the green fluorescence of 4T1 cell membrane overlap, and form yellow fluorescence, which proves the successful fusion of hybrid cell membrane HM.

[0065] (3) Immune escape ability of DDT-HM biomimetic nanoparticles:

[0066] When preparing DDT-HM biomimetic nanoparticles according to the scheme of Example 1, control DDT-CM and DDT-MM were set. DDT-CM and DDT-MM nanoparticles were respectively formed by DDT (DDT-Se) nanoparticles coated with 4T1 cell membrane alone and RAW cell membrane alone.

[0067] L929 cells, U87 cells, MCF-7 cells, RAW cells and 4T1 cells were cultured in 6-well plates for 18 hours. Then the culture medium was replaced with a DMEM solution containing DDT-HM, DDT-CM or DDT-MM. After incubation for 4 h, free materials were removed by PBS washing three times, and finally the cell uptake was quantitatively analyzed by FACS.

[0068] Results are shown in Figure 6 : The fluorescence intensity of DDT-HM and DDT-CM containing RAW membrane proteins in RAW cells is weak, which proves that the use of RAW membrane makes DDT-HM have the ability of immune escape.

[0069] (4) Optimal fusion membrane protein ratio of hybrid cell membrane:

[0070] RAW cell membranes labeled with CD206-specific markers and 4T1 cell membranes labeled with CD44-specific markers were extracted, and their membrane protein ratios were changed. After 1h of incubation, the labeled hybrid cell membranes HM were prepared by a liposome extruder, and finally FACS was used for quantitative analysis of cell uptake.

[0071] Results are shown in Figure 7 : When the mass ratio of 4T1 cell membrane protein: RAW cell membrane protein is 1:2, the fusion effect is best.

[0072] (5) Homologous targeting ability of DDT-HM biomimetic nanoparticles and optimal application membrane protein ratio:

[0073] L929 cells, U87 cells, MCF-7 cells, RAW cells and 4T1 cells were cultured in 6-well plates for 18 hours. Then the culture medium was replaced with DMEM solution containing DDT-HM (prepared according to the scheme of Example 1) with different membrane protein ratios. After 4h of incubation, PBS was washed three times to remove free materials, and finally FACS was used for quantitative analysis of cell uptake.

[0074] As shown in Figure 5 : The fluorescence intensity of DDT-HM biomimetic nanoparticles under any membrane protein ratio is the strongest in 4T1 cells, which proves that DDT-HM has the ability of homologous targeting. At the same time, the fluorescence intensity in RAW cells is moderate, which proves that DDT-HM has the ability of immune escape and homologous targeting at the same time, and the effect is best when the mass ratio of 4T1: RAW membrane protein is 3:1.

[0075] (6) Cell cytotoxicity detection of DDT-HM biomimetic nanoparticles

[0076] 4T1 cells were seeded in 96-well plates and incubated for 18 h. The culture medium was replaced with fresh DMEM containing free DDT-HM nanoparticles (DDT-HM concentration: 0.05, 0.10, 0.15, 0.20, 0.25 pg / mL, prepared as in Example 1). After 4 h of incubation, the cells were then irradiated with or without 808 nm NIR laser for 3 min. The cells were further incubated for 18 h, and the culture medium was replaced with MTT solution and incubated for another 4 h. Subsequently, the culture medium was removed and DMSO was added to each well to lyse the formazan crystals for 15 min. After the formazan crystals were completely dissolved, the absorbance (OD) at 570 nm was measured on a microplate reader. Cell survival rate was calculated according to the following formula, and the cell growth inhibition curve was plotted.

[0077]

[0078] The results are shown in Figure 8 : DDT-HM nanoparticles have good biocompatibility without laser irradiation and show excellent photothermal properties under 808 nm laser irradiation, effectively killing cancer cells.

[0079] (7) DDT-HM nanoparticles induce the expression of cell surface calreticulin (CRT)

[0080] 4T1 cells were incubated in 24-well plates for 24 h. DMEM solution containing DDT-HM nanoparticles was added to 4T1 cells and incubated for 4 h. Then, the cells were irradiated with or without 808 nm laser. Next, 4T1 cells were harvested, washed with pre-cooled PBS for 3 times, and then fixed in paraformaldehyde for 5 min. After washing in cold PBS, the cells were incubated with CRT primary antibody. After 30 min, the cells were washed again and incubated with Alexa Fluor 48-conjugated monoclonal secondary antibody for 30 min. Finally, the cells were stained with DAPI and analyzed by FACS.

[0081] The results are shown in Figure 9 : The relative fluorescence intensity of cells treated with DDT-HM nanoparticles + L (laser irradiation) was significantly higher than that of other groups, indicating that DDT-HM nanoparticles can induce high expression of CRT in cancer cells under laser irradiation. In the figure, DDT refers to DDT-Se.

[0082] (8) DDT-HM nanoparticles induce high mobility group box-1 (HMGB-1) efflux

[0083] 4T1 cells were seeded on cover slips placed in 24-well plates and incubated for 24 hours. DMEM solution containing DDT-HM biomimetic nanoparticles was added to 4T1 cells and incubated for 4 hours. Then irradiated with or without 808 nm laser. Next, intracellular HMGB-1 was stained with antibody. Cells were fixed with paraformaldehyde for 20 minutes, then permeabilized with Triton X-100. Non-specific binding sites were blocked by pre-incubation with 5% fetal bovine serum in PBS for 30 minutes, then incubated with HMGB-1 primary antibody for 1 hour, then incubated with Alexa Fluor 488 conjugated secondary antibody for 30 minutes, PBS washed. Finally, cells were stained with DAPI, observed by fluorescence microscopy and analyzed by FACS.

[0084] As shown in Figures 10-11 : obvious green fluorescence was observed in the cytoplasm of DDT-HM biomimetic nanoparticle + laser treated cells, proving that under the irradiation of laser CFIN can induce cancer cells to expel HMGB-1 on the nucleus to the cytoplasm, DDT in the figure is DDT-Se. At the same time, the relative fluorescence intensity of FACS analysis can prove this point.

[0085] (9) Anti-tumor efficacy of DDT-HM biomimetic nanoparticles in mice

[0086] Balb / c mice were subcutaneously implanted with 4T1 cancer cells, and dosing began seven days later (wherein DDT-HM, DDT is DDT-Se, the concentration is 5 μg / mL, the preparation means is as described in Example 1), dosing once every other day, a total of 6 times. The primary tumor of each mouse was irradiated with 808 nm laser for 10 minutes 4 hours after dosing. The maximum diameter and minimum diameter of the tumor of the tumor-bearing mouse were measured every two days from the start of dosing to record the tumor size.

[0087] As shown in Figure 12 : under the irradiation of laser, the tumor growth of DDT-HM biomimetic nanoparticle treated mice was significantly inhibited.

[0088] The above examples have described the technical solutions of the present application in detail, it should be understood that the above description is only a specific embodiment of the present application, and is not used to limit the present application, any modification, supplement or similar way of substitution made within the principle range of the present application, should be included in the protection scope of the present application.

Claims

1. A hybrid cell membrane-modified biomimetic nanoparticle having an antitumor effect, characterized by, The hybrid cell membrane comprises a nanoparticle core and a hybrid cell membrane covering the nanoparticle core. The nanoparticle core comprises an amphiphilic polymer drug carrier and an active drug, and the active drug comprises a photothermal agonist TPT and a STING agonist DMXAA; the mass ratio of the photothermal agonist TPT, the STING agonist DMXAA and the amphiphilic polymer drug carrier is 1:10:10-15. The photothermal agonist TPT has the following structural formula: The STING agonist DMXAA has the following structural formula: The hybrid cell membrane is obtained by co-extrusion of a cancer cell membrane and a macrophage cell membrane.

2. The hybrid cell-membrane modified, bi-nanoparticle having an anti-tumor effect according to claim 1, wherein, The amphiphilic polymer drug carrier is distearoyl phosphatidyl ethanolamine-polyethylene glycol.

3. The hybrid cell-membrane modified, bi-nanoparticle having an anti-tumor effect according to claim 1, wherein, The amphiphilic polymeric drug carrier is DSPE-PEG 2000 .

4. The hybrid cell-membrane modified, bi-nanoparticle with anti-tumor effect according to claim 1, wherein, The hybrid cell membrane is obtained by co-extrusion of a 4T1 cell membrane and a RAW cell membrane, the 4T1 cell membrane is the cell membrane of 4T1 mouse breast cancer cells, and the RAW cell membrane is the cell membrane of RAW264.7 mouse mononuclear macrophages.

5. The hybrid cell-membrane modified, bi-nanoparticle with anti-tumor effect according to claim 4, wherein, The co-extrusion is performed according to the mass ratio of the membrane proteins of the 4T1 cell membrane and the RAW cell membrane being 1-3:

1.

6. The method of claim 1-5, wherein the preparation method of the hybrid cell membrane modified bionanoparticle having an antitumor effect is characterized by, The method comprises the following steps: Step S1: under the action of ice bath and ultrasonic, an organic solution of the photothermal agonist TPT and the STING agonist DMXAA is added dropwise into an aqueous solution of the amphiphilic polymer, so that the amphiphilic polymer coats the photothermal agonist TPT and the STING agonist DMXAA, and then the organic solvent and water are removed to obtain a nanoparticle core DDT; Step S2: the cancer cell membrane and the macrophage cell membrane are co-extruded in a liposome extruder to obtain a hybrid cell membrane; Step S3: the nanoparticle core is co-extruded with the hybrid cell membrane to obtain the hybrid cell membrane modified bionanoparticle with anti-tumor effect.

7. The method for preparing hybrid cell membrane-modified biomimetic nanoparticles with antitumor activity according to claim 6, characterized in that, The organic solvent in the organic solution of the photothermal agonist TPT and the STING agonist DMXAA is tetrahydrofuran.

8. The method for preparing hybrid cell membrane-modified biomimetic nanoparticles with antitumor activity according to claim 6, characterized in that, The cancer cell membrane is prepared by centrifuging to collect cell precipitate, resuspending in a hypotonic lysis buffer, incubating in an ice bath, repeatedly freezing and thawing, collecting the supernatant by low-speed centrifugation, and collecting the precipitate by high-speed centrifugation of the obtained supernatant; and the macrophage cell membrane is obtained by treating macrophages in the same way.

9. The use of the hybrid cell membrane modified bionanoparticle with anti-tumor effect according to any one of claims 1-5 in the preparation of a photothermal activation immunotherapy preparation for tumors.

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