Bionic nanoparticles based on cancer cell membrane modification and preparation method and application thereof

Abstract

The invention discloses a bionic nanoparticle based on cancer cell membrane modification and a preparation method and application thereof, belongs to the technical field of nano drug loading, and particularly relates to a bionic nanoparticle based on cancer cell membrane modification, which comprises a carrier, a utility agent loaded on the carrier and a cancer cell membrane coated on the outer layer, the carrier contains CeO2 and MoS2, the CeO2 is of a hollow structure, and the MoS2 is combined on the hollow structure of the CeO2. The bionic nanoparticles based on cancer cell membrane modification prepared by the invention are good in targeting, good in anti-tumor activity, low in cytotoxicity, good in tumor growth inhibition and tumor targeting accumulation capacity, long in circulation time and capable of effectively killing tumor cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of nano-drug delivery, and particularly relates to biomimetic nanoparticles modified with cancer cell membranes, and a preparation method and application thereof. Background Art

[0002] Breast cancer is a common malignant tumor and the second leading cause of cancer-related deaths, seriously threatening the lives and health of women. The treatment methods for breast cancer mainly include surgery, radiotherapy, chemotherapy, endocrine therapy, targeted therapy, immunotherapy, etc., but breast cancer still has a relatively high recurrence rate and mortality rate. In recent years, the research on multi-modal combination therapy has gradually deepened. Among them, chemoimmunotherapy applies the cytotoxic and immunomodulatory effects of chemotherapeutic drugs, and is a treatment method that combines immunotherapy strategies to synergistically fight tumors, with broad application prospects. Although chemoimmunotherapy shows great potential in cancer treatment, its core challenge still focuses on how to enhance the synergistic treatment effect while effectively reducing the side effects caused by multiple drugs. In the past few decades, the drug delivery system has emerged as a cutting-edge strategy to address this challenge. As a new carrier for chemoimmunotherapy, it can significantly extend the drug action time, promote the accumulation of drugs in the lesion area, and accurately target specific treatment targets, greatly avoiding potential harm to non-target tissues. This progress has opened up a more precise and personalized treatment approach for the chemoimmunotherapy of breast cancer.

[0003] Molybdenum disulfide (MoS2), as a layered transition metal sulfide, has a large specific surface area. After modification and modification, MoS2 can load drug molecules or immune adjuvants to achieve targeted delivery and controlled release. In addition, MoS2 can generate local high temperatures under near-infrared light irradiation, thereby triggering immunogenic cell death (ICD) of tumor cells and enhancing the immunotherapy effect. However, compared with other nanomaterials such as gold nanoparticles, the photothermal conversion efficiency of molybdenum disulfide nanosheets is relatively low, which to a certain extent limits its application in cancer-photothermal therapy; at the same time, the clearance by the immune system, poor antigen presentation in tumor tissues, and the lack of specific targets are also the biggest obstacles restricting the MoS2 nano-drug delivery system at present. Therefore, it is particularly important to develop new MoS2-based nano-drug delivery immunopreparation system materials. Summary of the Invention

[0004] The purpose of the present invention is to provide biomimetic nanoparticles modified with cancer cell membranes, and a preparation method and application thereof, which have good targeting, good anti-tumor activity, low cytotoxicity, can inhibit tumor growth, have good tumor-targeting accumulation ability, long circulation time, and can effectively kill tumor cells.

[0005] The technical solution adopted by the present invention to achieve the above purpose is as follows: A biomimetic nanoparticle based on cancer cell membrane modification, comprising: a carrier, an agent loaded on the carrier, and an outer coating of cancer cell membrane; the carrier contains CeO2 and MoS2, CeO2 has a hollow structure, and MoS2 is combined with the hollow structure of CeO2. In the present invention, a hybrid nanostructure containing CeO2 and MoS2 is first prepared as a carrier by a chemical synthesis method, and then PTX is embedded into the carrier by a mechanical stirring method to further obtain nanoparticles with controllable drug release; subsequently, a PD-L1 inhibitor is coupled to the surface of the nanoparticles through an amidation reaction; finally, the extracted 4T1 cancer cell membrane is wrapped outside the above product by a mechanical extrusion technique to obtain a biomimetic nanoparticle (CPPN) based on cancer cell membrane modification that can target drug release. The CPPN nanoparticles provided by the present invention, in which the carrier containing the CeO2 and MoS2 hybrid nanostructure can not only provide a framework for drug loading but also is a photothermal agent. PTX, as a common chemotherapy drug for breast cancer, is combined with the PD-L1 inhibitor for immunotherapy. Wrapping the 4T1 cell membrane outside the nanoparticles can improve the effect of the biomimetic nanoparticles based on cancer cell membrane modification prepared in the present invention. The present invention provides an application basis for a nano-drug delivery immunopreparation system material containing a CeO2 and MoS2 hybrid nanostructure as a carrier, and also provides a basis for applying the above drug delivery system to the treatment of breast cancer.

[0006] Preferably, the agent includes at least one of PTX and PDL1i, and PDL1i is a PDL1 antibody; or, the cancer cell membrane is a 4T1 cell membrane.

[0007] The present invention discloses a preparation method of a biomimetic nanoparticle based on cancer cell membrane modification, comprising: S1, reacting CeO2 hollow spheres with sodium molybdate in a carrier reaction solution to form a carrier; the carrier reaction solution also contains thioacetamide; S2, treating the carrier with an agent solution to form a carrier-agent loaded with the agent; the agent solution contains the agent, and the agent includes at least one of PTX and PDL1i, and PDL1i is a PDL1 antibody; S3, mixing the carrier-agent with a cancer cell membrane suspension to prepare a biomimetic nanoparticle based on cancer cell membrane modification, and the cancer cell membrane suspension contains a cancer cell membrane.

[0008] Preferably, the carrier reaction solution is prepared by dispersing CeO2 hollow spheres in deionized water, and then adding sodium molybdate and thioacetamide; the usage amount of CeO2 hollow spheres is 0.05-0.2 wt% of deionized water, the usage amount of sodium molybdate is 200-300 wt% of CeO2 hollow spheres, and the usage amount of thioacetamide is 400-600 wt% of CeO2 hollow spheres; or, The carrier reaction solution is prepared by dispersing CeO2 hollow spheres in deionized water, and then adding sodium molybdate, thioacetamide, and polyethylene glycol monoester. The polyethylene glycol monoester is prepared by reacting polyethylene glycol with fatty acid. The usage amount of CeO2 hollow spheres is 0.05 - 0.2 wt% of deionized water, the usage amount of sodium molybdate is 200 - 300 wt% of CeO2 hollow spheres, the usage amount of thioacetamide is 400 - 600 wt% of CeO2 hollow spheres, and the usage amount of polyethylene glycol monoester is 20 - 100 wt% of sodium molybdate dihydrate. In the process of binding MoS2 to CeO2 hollow spheres in the present invention, polyethylene glycol monoester prepared by reacting polyethylene glycol with fatty acid is introduced, so that the obtained carrier has the structure of polyethylene glycol monoester. Through the structure of polyethylene glycol monoester and the structures of MoS2 and CeO2, and under the action of the frameworks of CeO2 and MoS2 and the photothermal agent, the introduction of the structure of polyethylene glycol monoester can better exert the effect of the drug and improve the therapeutic effect on tumors. The usage amount of polyethylene glycol monoester should not be too small, otherwise it will not be able to exert the effect. At the same time, if the usage amount of polyethylene glycol monoester is too high, it will hinder the exertion of the drug therapeutic effect. Therefore, the usage amount of polyethylene glycol monoester needs to be within a certain range to have a good effect.

[0009] Preferably, the fatty acid is a saturated hydrocarbon acid or an unsaturated hydrocarbon acid with C3 - 12.

[0010] More preferably, the fatty acid is caproic acid.

[0011] Preferably, the utility agent solution includes PTX solution and PDL1i-containing solution; in the preparation of the carrier-utility agent, first, the carrier is mixed with the PTX solution to obtain carrier@PTX, and then carrier@PTX is mixed with the PDL1i-containing solution to obtain carrier@PTX@PDL1i, that is, the carrier-utility agent.

[0012] Preferably, the PTX solution is prepared by mixing PTX and deionized water. The PTX solution contains 0.05 - 0.2 wt% of PTX. The PTX solution is measured by PTX, and the usage amount of the carrier is 100 - 300 wt% of PTX.

[0013] Preferably, the cancer cell membrane suspension is prepared by resuspending the cancer cell membrane in PBS solution containing PMSF. The pH of the PBS solution is 7 - 7.8; the PBS solution contains 0.5 - 2 mM of PMSF, and the usage amount relationship between the cancer cell membrane and the PBS solution is 0.1 - 2 mg / mL.

[0014] Preferably, the usage amount of the cancer cell membrane is 80 - 120 wt% of the carrier-utility agent.

[0015] The present invention discloses the use of the biomimetic nanoparticles modified based on cancer cell membranes prepared by the above preparation method in the preparation of anti-tumor drugs.

[0016] Preferably, in the preparation of CeO2 hollow spheres, ammonium hydroxide solution is added to the tetraethyl orthosilicate mixture, stirred at 20 - 40 °C for 12 - 48 h, centrifuged, dried to obtain SiO2. Then, SiO2 is dispersed in ethylene glycol, cerium nitrate hexahydrate and deionized water are added, stirred and mixed evenly, transferred to a reaction kettle, and heat-treated at 120 - 160 °C for 12 - 48 h, centrifuged, washed and dried to obtain SiO2@CeO2 particles. Finally, it is dispersed in an alkaline solution for 1 - 4 d, centrifuged, washed and dried to obtain CeO2 hollow spheres.

[0017] More preferably, in the preparation of CeO2 hollow spheres, the ammonium hydroxide solution is composed of ammonium hydroxide and deionized water, and the ammonium hydroxide solution contains 6 - 18 wt% ammonium hydroxide.

[0018] More preferably, in the preparation of CeO2 hollow spheres, the tetraethyl orthosilicate mixture is composed of tetraethyl orthosilicate and ethanol, and the tetraethyl orthosilicate mixture contains 1 - 5 wt% tetraethyl orthosilicate.

[0019] More preferably, in the preparation of CeO2 hollow spheres, when the ammonium hydroxide solution is mixed with the tetraethyl orthosilicate mixture, the ammonium hydroxide solution is measured by ammonium hydroxide, the tetraethyl orthosilicate mixture is measured by tetraethyl orthosilicate, and the usage amount of ammonium hydroxide is 80 - 120 wt% of tetraethyl orthosilicate.

[0020] More preferably, in the preparation of CeO2 hollow spheres, the usage amount of SiO2 is 0.3 - 2.4 wt% of ethylene glycol.

[0021] More preferably, in the preparation of CeO2 hollow spheres, the usage amount of cerium nitrate hexahydrate is 400 - 600 wt% of SiO2.

[0022] More preferably, in the preparation of CeO2 hollow spheres, the usage amount of deionized water is 700 - 800 wt% of SiO2, the alkaline solution is sodium hydroxide solution, and the concentration of sodium hydroxide in the sodium hydroxide solution is 2 - 6 M.

[0023] Preferably, in the preparation of CeO2-MoS2 hybrid nanocarriers, CeO2 hollow spheres are dispersed in deionized water, ultrasonically treated for 10 - 60 min, then sodium molybdate and thioacetamide are added, stirred for 10 - 60 min, then transferred to a reaction kettle, treated at 190 - 220 °C for 12 - 48 h, centrifuged, washed and dried to obtain CeO2-MoS2 hybrid nanocarriers.

[0024] More preferably, in the preparation of CeO2-MoS2 hybrid nanocarriers, the usage amount of CeO2 hollow spheres is 0.05 - 0.2 wt% of deionized water.

[0025] More preferably, in the preparation of the CeO2-MoS2 hybrid nanocarrier, the usage amount of sodium molybdate is 200-300 wt% of the CeO2 hollow spheres.

[0026] More preferably, in the preparation of the CeO2-MoS2 hybrid nanocarrier, the usage amount of thioacetamide is 400-600 wt% of the CeO2 hollow spheres.

[0027] Preferably, in the preparation of polyethylene glycol monoester, polyethylene glycol and fatty acid are added into xylene, then a catalyst is added, and the reaction is carried out at 150-170 °C for 3-12 h. After the reaction is completed, it is washed successively with toluene, isopropyl alcohol and saturated brine, and vacuum dried to obtain polyethylene glycol monoester.

[0028] More preferably, in the preparation of polyethylene glycol monoester, the molecular weight of polyethylene glycol is 100-500, and the usage amount of polyethylene glycol is 10-50 wt% of xylene.

[0029] More preferably, in the preparation of polyethylene glycol monoester, the fatty acid is hexanoic acid, and the usage molar amount of the fatty acid is 60-100% of the usage molar amount of polyethylene glycol.

[0030] More preferably, in the preparation of polyethylene glycol monoester, the catalyst is zirconium butoxide solution, which is mixed by zirconium butoxide and isopropyl alcohol. The content of zirconium butoxide in the zirconium butoxide solution is 0.05-0.3 wt%, and the usage amount of the catalyst is 0.5-3 wt% of polyethylene glycol.

[0031] Preferably, in the preparation of the CeO2-MoS2 hybrid composite nanocarrier, the CeO2 hollow spheres are dispersed in deionized water, ultrasonically treated for 10-60 min, then sodium molybdate and thioacetamide are added, stirred for 10-60 min, then polyethylene glycol monoester is added and mixed, then transferred to a reaction kettle, treated at 190-220 °C for 12-48 h, centrifuged, washed and dried to obtain the CeO2-MoS2 hybrid nanocarrier.

[0032] More preferably, in the preparation of the CeO2-MoS2 hybrid composite nanocarrier, the usage amount of the CeO2 hollow spheres is 0.05-0.2 wt% of deionized water.

[0033] More preferably, in the preparation of the CeO2-MoS2 hybrid composite nanocarrier, the usage amount of sodium molybdate is 200-300 wt% of the CeO2 hollow spheres.

[0034] More preferably, in the preparation of the CeO2-MoS2 hybrid composite nanocarrier, the usage amount of thioacetamide is 400-600 wt% of the CeO2 hollow spheres.

[0035] More preferably, in the preparation of the CeO2-MoS2 hybrid composite nanocarrier, the usage amount of polyethylene glycol monoester is 20-100 wt% of sodium molybdate dihydrate.

[0036] Preferably, in the preparation of CeO2-MoS2@PTX, the CeO2-MoS2 hybrid nanocarrier is added to the PTX solution, stirred for 10-48 h under light-shielded conditions, centrifuged, washed, and freeze-dried to obtain CeO2-MoS2@PTX.

[0037] More preferably, in the preparation of CeO2-MoS2@PTX, the PTX solution is composed of PTX and deionized water, and the PTX solution contains 0.05-0.2 wt% of PTX.

[0038] More preferably, in the preparation of CeO2-MoS2@PTX, the PTX solution is measured by PTX, and the usage amount of the CeO2-MoS2 hybrid nanocarrier is 100-300 wt% of PTX.

[0039] Preferably, in the preparation of CeO2-MoS2@PTX, the CeO2-MoS2 hybrid nanocarrier and L-rhamnose diethyl mercaptal are added to the PTX solution, stirred for 10-48 h under light-shielded conditions, centrifuged, washed, and freeze-dried to obtain CeO2-MoS2@PTX. When loading PTX in the present invention, L-rhamnose diethyl mercaptal can also be added to the drug, and L-rhamnose diethyl mercaptal and PTX are co-loaded in the CeO2-MoS2 hybrid nanocarrier, so that in addition to PTX, CeO2-MoS2@PTX also contains L-rhamnose diethyl mercaptal. The use of L-rhamnose diethyl mercaptal can improve the therapeutic effect of the cancer cell membrane-based biomimetic nanoparticles on tumors under the action of the components of other cancer cell membrane-based biomimetic nanoparticles.

[0040] More preferably, in the preparation of CeO2-MoS2@PTX, the PTX solution is composed of PTX and deionized water, and the PTX solution contains 0.05-0.2 wt% of PTX.

[0041] More preferably, in the preparation of CeO2-MoS2@PTX, the PTX solution is measured by PTX, and the usage amount of the CeO2-MoS2 hybrid nanocarrier is 100-300 wt% of PTX.

[0042] More preferably, in the preparation of CeO2-MoS2@PTX, the usage amount of L-rhamnose diethyl mercaptal is 3-21 wt% of PTX.

[0043] Preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, CeO2-MoS2@PTX, EDC·HCl and NHS are added to a PBS solution, ultrasonicated for 10 - 60 min, then PDL1 antibody is added, and stirred in the dark at 20 - 40 °C for 10 - 48 h to obtain CeO2-MoS2@PTX@PDL1i.

[0044] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the pH of the PBS solution is 7 - 7.8.

[0045] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the usage amount of CeO2-MoS2@PTX is 0.05 - 0.4 wt% of the PBS solution.

[0046] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the usage amount of EDC·HCl is 100 - 300 wt% of CeO2-MoS2@PTX.

[0047] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the usage amount of NHS is 100 - 300 wt% of CeO2-MoS2@PTX.

[0048] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the usage amount of PDL1 antibody is 1 - 10 wt% of CeO2-MoS2@PTX.

[0049] Preferably, in the extraction of cancer cell membranes, 4T1 cells are cultured in DMEM medium. After the 4T1 cells grow to confluence, the 4T1 cells are taken out, resuspended with a PBS solution, the precipitate is collected by centrifugation, resuspended in a hypotonic lysis buffer, and treated on ice bath for 5 - 30 min. Then the precipitate is repeatedly frozen and thawed 2 - 5 times in liquid nitrogen and a water bath at 30 - 40 °C, centrifuged at 500 - 1000 g at 0 - 10 °C for 5 - 15 min, the supernatant is collected, centrifuged again for 10 - 60 min, the precipitate is collected, resuspended with a hypotonic lysis buffer, and centrifuged again at 10000 - 18000 g at 0 - 10 °C for 10 - 60 min to obtain cancer cell membranes.

[0050] More preferably, in the extraction of cancer cell membranes, the DMEM medium contains 5 - 15% FBS, and the DMEM medium also contains penicillin and streptomycin. The amount of penicillin is 20 - 100 IU / mL, and the amount of streptomycin is 20 - 100 IU / mL.

[0051] More preferably, in the extraction of cancer cell membranes, the pH of the PBS solution is 7 - 7.8.

[0052] More preferably, in the extraction of cancer cell membranes, the hypotonic lysis buffer is a PBS solution containing PMSF, and the content of PMSF in the hypotonic lysis buffer is 0.1 - 5 mM.

[0053] More preferably, in the extraction of cancer cell membranes, during the resuspension treatment, the usage amount of the solid matter and the liquid reagent in the resuspension is 0.1 - 1 mg / mL.

[0054] Preferably, in the preparation of CeO2-MoS2@PTX biomimetic nanoparticles, the cancer cell membranes are resuspended in a PBS solution containing PMSF, then CeO2-MoS2@PTX@PDL1i is added and mixed, and after coating with a polycarbonate membrane and mechanical extrusion, it is then bath sonicated for 10 - 60 min under dark conditions at 0 - 10 °C to obtain CeO2-MoS2@PTX biomimetic nanoparticles.

[0055] More preferably, in the preparation of CeO2-MoS2@PTX biomimetic nanoparticles, the pH of the PBS solution is 7 - 7.8.

[0056] More preferably, in the preparation of CeO2-MoS2@PTX biomimetic nanoparticles, the PBS solution contains 0.5 - 2 mM of PMSF, and the usage amount relationship between the cancer cell membranes and the PBS solution is 0.1 - 2 mg / mL.

[0057] More preferably, in the preparation of CeO2-MoS2@PTX biomimetic nanoparticles, the usage amount of the cancer cell membranes is 80 - 120 wt% of CeO2-MoS2@PTX@PDL1i.

[0058] The present invention discloses an anti-tumor drug, comprising the above-mentioned biomimetic nanoparticles modified based on cancer cell membranes.

[0059] Since the present invention adopts a biomimetic nanoparticle modified based on cancer cell membrane, which is formed by a carrier, an effector loaded on the carrier, and an outer coating of cancer cell membrane, the carrier is composed of CeO2 and MoS2, CeO2 has a hollow structure, and then MoS2 is combined through synthesis, and then an effector including PTX and PDL1i is further loaded, and finally the cancer cell membrane is coated; when preparing the carrier, when preparing the carrier combined with MoS2 from hollow CeO2, polyethylene glycol monoester can also be added, and polyethylene glycol monoester is made by reacting polyethylene glycol with fatty acid, thus having the following beneficial effects: the biomimetic nanoparticle modified based on cancer cell membrane prepared by the present invention has good targeting, good anti-tumor activity, low cytotoxicity, can inhibit tumor growth, has the ability of tumor-targeted accumulation, long circulation time, and can effectively kill tumor cells. Therefore, the present invention is a biomimetic nanoparticle modified based on cancer cell membrane, its preparation method and application, which have good targeting, good anti-tumor activity, low cytotoxicity, can inhibit tumor growth, has the ability of tumor-targeted accumulation, long circulation time, and can effectively kill tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 It is an electron micrograph of the biomimetic nanoparticle.

[0061] Figure 2 It is a result graph of in vitro homologous targeting detection.

[0062] Figure 3 It is a result graph of in vitro anti-tumor activity detection.

[0063] Figure 4 It is a result graph of in vitro anti-tumor activity detection of the biomimetic nanoparticle modified based on cancer cell membrane.

[0064] Figure 5 It is a result graph of in vitro cytotoxicity test.

[0065] Figure 6 It is a result graph of in vivo anti-tumor performance test.

[0066] Figure 7 It is an in vivo fluorescence imaging graph. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0067] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0068] First, the concepts involved in this application will be described in conjunction with the accompanying drawings. It should be noted here that the following descriptions of each concept are only for making the content of this application easier to understand and do not represent a limitation on the protection scope of this application; at the same time, without conflict, the embodiments and features in the embodiments of this application can be combined with each other. This application will be described in detail below with reference to the accompanying drawings and in conjunction with the embodiments.

[0069] Example 1: A preparation method of biomimetic nanoparticles based on cancer cell membrane modification Preparation of CeO2 hollow spheres: Add ammonium hydroxide solution to the tetraethyl orthosilicate mixture, stir at 25 °C for 24 h, centrifuge and separate, and dry to obtain SiO2. Then disperse SiO2 in ethylene glycol, add cerium nitrate hexahydrate and deionized water, stir and mix evenly, transfer to a reaction kettle, and heat-treat at 130 °C for 24 h. After centrifugation, wash and dry to obtain SiO2@CeO2 particles. Finally, disperse in an alkaline solution for 2 d, centrifuge, wash and dry to obtain CeO2 hollow spheres. The ammonium hydroxide solution is formed by mixing ammonium hydroxide and deionized water, and the ammonium hydroxide solution contains 13 wt% ammonium hydroxide. The tetraethyl orthosilicate mixture is formed by mixing tetraethyl orthosilicate and ethanol, and the tetraethyl orthosilicate mixture contains 2.78 wt% tetraethyl orthosilicate. When the ammonium hydroxide solution is mixed with the tetraethyl orthosilicate mixture, the ammonium hydroxide solution is measured by ammonium hydroxide, and the tetraethyl orthosilicate mixture is measured by tetraethyl orthosilicate. The usage amount of ammonium hydroxide is 105 wt% of tetraethyl orthosilicate; the usage amount of SiO2 is 0.77 wt% of ethylene glycol, the usage amount of cerium nitrate hexahydrate is 500 wt% of SiO2, the usage amount of deionized water is 750 wt% of SiO2, the alkaline solution is sodium hydroxide solution, and the concentration of sodium hydroxide in the sodium hydroxide solution is 5 M.

[0070] Preparation of CeO2-MoS2 hybrid nanocarriers: Disperse CeO2 hollow spheres in deionized water, ultrasonically treat for 30 min, then add sodium molybdate and thioacetamide, stir for 30 min, then transfer to a reaction kettle, and treat at 200 °C for 24 h. After centrifugation, wash and dry to obtain CeO2-MoS2 hybrid nanocarriers. The usage amount of CeO2 hollow spheres is 0.14 wt% of deionized water, the usage amount of sodium molybdate is 250 wt% of CeO2 hollow spheres, and the usage amount of thioacetamide is 500 wt% of CeO2 hollow spheres.

[0071] Preparation of CeO2-MoS2@PTX: Add the CeO2-MoS2 hybrid nanocarrier into the PTX solution, stir for 24 h under dark conditions, wash after centrifugation, and freeze-dry to obtain CeO2-MoS2@PTX. The PTX solution is composed of PTX and deionized water. The PTX solution contains 0.1 wt% of PTX. The PTX solution is measured by PTX, and the usage amount of the CeO2-MoS2 hybrid nanocarrier is 200 wt% of PTX.

[0072] Preparation of CeO2-MoS2@PTX@PDL1i: Add CeO2-MoS2@PTX, EDC·HCl, and NHS into the PBS solution, ultrasonically treat for 20 min, then add the PDL1 antibody, and stir under dark conditions at 25 °C for 24 h to obtain CeO2-MoS2@PTX@PDL1i. The pH of the PBS solution is 7.4. The usage amount of CeO2-MoS2@PTX is 0.3 wt% of the PBS solution. The usage amount of EDC·HCl is 200 wt% of CeO2-MoS2@PTX. The usage amount of NHS is 200 wt% of CeO2-MoS2@PTX. The usage amount of the PDL1 antibody is 5 wt% of CeO2-MoS2@PTX.

[0073] Extraction of cancer cell membrane: Culture 4T1 cells in DMEM medium. After the 4T1 cells grow to confluence, take out the 4T1 cells, resuspend them with PBS solution, centrifuge to collect the precipitate, resuspend it in the hypotonic lysis buffer, and treat it in an ice bath for 100 min. Then, repeatedly freeze-thaw the precipitate 3 times in liquid nitrogen and a 37 °C water bath, centrifuge at 700 g for 10 min at 4 °C, collect the supernatant, centrifuge again for 30 min, collect the precipitate, add the hypotonic lysis buffer to resuspend it, and centrifuge again at 14000 g for 30 min at 4 °C to obtain the cancer cell membrane. The DMEM medium contains 10% FBS. The DMEM medium also contains penicillin and streptomycin. The amount of penicillin is 50 IU / mL, and the amount of streptomycin is 50 IU / mL. The pH of the PBS solution is 7.4. The hypotonic lysis buffer is a PBS solution containing PMSF, and the content of PMSF in the hypotonic lysis buffer is 1 mM. In the resuspension treatment, the usage amount of the solid matter and the liquid reagent in the resuspension is 0.5 mg / mL.

[0074] Preparation of CeO2-MoS2@PTX biomimetic nanoparticles: The cancer cell membrane was resuspended in a PBS solution containing PMSF, and then CeO2-MoS2@PTX@PDL1i was added and mixed. After coating with a polycarbonate membrane and mechanical extrusion, it was then bath sonicated for 20 min in the dark at 4 °C to obtain CeO2-MoS2@PTX biomimetic nanoparticles. The pH of the PBS solution was 7.4; the PBS solution contained 1 mM of PMSF, the usage relationship between the cancer cell membrane and the PBS solution was 0.5 mg / mL, and the usage amount of the cancer cell membrane was 100 wt% of CeO2-MoS2@PTX@PDL1i.

[0075] Example 2: A method for preparing a biomimetic nanoparticle modified with a cancer cell membrane Compared with Example 1, the difference in this example is that in the preparation of CeO2-MoS2@PTX, the CeO2-MoS2 hybrid nanocarrier was replaced with a CeO2-MoS2 hybrid composite nanocarrier.

[0076] Preparation of polyethylene glycol monoester: Polyethylene glycol and fatty acid were added to xylene, and then a catalyst was added. The reaction was carried out at 160 °C for 6 h. After the reaction was completed, it was washed successively with toluene, isopropanol and saturated brine, and vacuum dried to obtain polyethylene glycol monoester. The molecular weight of polyethylene glycol was 300, the usage amount of polyethylene glycol was 30 wt% of xylene, the fatty acid was hexanoic acid, the molar usage amount of the fatty acid was 80% of the molar usage amount of polyethylene glycol, the catalyst was a zirconium butoxide solution, the zirconium butoxide solution was composed of zirconium butoxide and isopropanol, the content of zirconium butoxide in the zirconium butoxide solution was 0.15 wt%, and the usage amount of the catalyst was 1 wt% of polyethylene glycol.

[0077] Preparation of CeO2-MoS2 hybrid composite nanocarrier: CeO2 hollow spheres were dispersed in deionized water and sonicated for 30 min, then sodium molybdate and thioacetamide were added, stirred for 30 min, then polyethylene glycol monoester was added and mixed, and then transferred to a reaction kettle and treated at 200 °C for 24 h. After centrifugation, it was washed and dried to obtain a CeO2-MoS2 hybrid nanocarrier. The usage amount of CeO2 hollow spheres was 0.14 wt% of deionized water, the usage amount of sodium molybdate was 250 wt% of CeO2 hollow spheres, the usage amount of thioacetamide was 500 wt% of CeO2 hollow spheres, and the usage amount of polyethylene glycol monoester was 40 wt% of sodium molybdate dihydrate.

[0078] Example 3: A method for preparing a biomimetic nanoparticle modified with a cancer cell membrane Compared with Example 2, the difference lies in the preparation of the CeO2-MoS2 hybrid composite nanocarrier.

[0079] Preparation of CeO2-MoS2 Hybrid Composite Nanocarriers: Disperse CeO2 hollow spheres in deionized water, ultrasonically treat for 30 min, then add sodium molybdate and thioacetamide, stir for 30 min, then add polyethylene glycol monoester and mix, then transfer to a reaction kettle, treat at 200 °C for 24 h, centrifuge, wash and dry to obtain CeO2-MoS2 hybrid nanocarriers. The usage amount of CeO2 hollow spheres is 0.14 wt% of deionized water, the usage amount of sodium molybdate is 250 wt% of CeO2 hollow spheres, the usage amount of thioacetamide is 500 wt% of CeO2 hollow spheres, and the usage amount of polyethylene glycol monoester is 90 wt% of sodium molybdate dihydrate.

[0080] Example 4: A preparation method of biomimetic nanoparticles based on cancer cell membrane modification Compared with Example 2, the difference in this example lies in CeO2-MoS2@PTX.

[0081] Preparation of CeO2-MoS2@PTX: Add CeO2-MoS2 hybrid nanocarriers and L-rhamnose diethyl mercaptal to the PTX solution, stir in the dark for 24 h, centrifuge, wash, and freeze-dry to obtain CeO2-MoS2@PTX. The PTX solution is composed of PTX and deionized water, the PTX solution contains 0.1 wt% of PTX, the PTX solution is measured by PTX, the usage amount of CeO2-MoS2 hybrid nanocarriers is 200 wt% of PTX, and the usage amount of L-rhamnose diethyl mercaptal is 5 wt% of PTX.

[0082] Example 5: A preparation method of biomimetic nanoparticles based on cancer cell membrane modification Compared with Example 2, the difference in this example lies in that in the preparation of CeO2-MoS2@PTX@PDL1i, CeO2-MoS2@PTX is replaced by CeO2-MoS2@PTX-L.

[0083] Preparation of CeO2-MoS2@PTX-L: Add CeO2-MoS2 hybrid nanocarriers and L-rhamnose diethyl mercaptal to the PTX solution, stir in the dark for 10 - 48 h, centrifuge, wash, and freeze-dry to obtain CeO2-MoS2@PTX. The PTX solution is composed of PTX and deionized water, the PTX solution contains 0.05 - 0.2 wt% of PTX, the PTX solution is measured by PTX, the usage amount of CeO2-MoS2 hybrid nanocarriers is 100 - 300 wt% of PTX, and the usage amount of L-rhamnose diethyl mercaptal is 15 wt% of PTX.

[0084] Comparative Example 1: A preparation method of biomimetic nanoparticles based on cancer cell membrane modification This comparative example is different from Example 2 in the preparation of the CeO2-MoS2 hybrid composite nanocarrier.

[0085] Preparation of the CeO2-MoS2 hybrid composite nanocarrier: Disperse CeO2 hollow spheres in deionized water, sonicate for 30 min, then add sodium molybdate and thioacetamide, stir for 30 min, then add polyethylene glycol monoester and mix. Then transfer to a reaction kettle and treat at 200 °C for 24 h. After centrifugation, wash and dry to obtain the CeO2-MoS2 hybrid nanocarrier. The usage amount of CeO2 hollow spheres is 0.14 wt% of deionized water, the usage amount of sodium molybdate is 250 wt% of CeO2 hollow spheres, the usage amount of thioacetamide is 500 wt% of CeO2 hollow spheres, and the usage amount of polyethylene glycol monoester is 10 wt% of sodium molybdate dihydrate.

[0086] Comparative Example 2: A preparation method of biomimetic nanoparticles modified with cancer cell membranes This comparative example is different from Example 2 in the preparation of the CeO2-MoS2 hybrid composite nanocarrier.

[0087] Preparation of the CeO2-MoS2 hybrid composite nanocarrier: Disperse CeO2 hollow spheres in deionized water, sonicate for 30 min, then add sodium molybdate and thioacetamide, stir for 30 min, then add polyethylene glycol monoester and mix. Then transfer to a reaction kettle and treat at 200 °C for 24 h. After centrifugation, wash and dry to obtain the CeO2-MoS2 hybrid nanocarrier. The usage amount of CeO2 hollow spheres is 0.14 wt% of deionized water, the usage amount of sodium molybdate is 250 wt% of CeO2 hollow spheres, the usage amount of thioacetamide is 500 wt% of CeO2 hollow spheres, and the usage amount of polyethylene glycol monoester is 120 wt% of sodium molybdate dihydrate.

[0088] Test example: The CeO2-MoS2@PTX@PD-L1i and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1 of the present invention were used as representatives for research and testing.

[0089] The CeO2-MoS2@PTX@PD-L1i and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1 of the present invention were characterized by scanning electron microscopy, and the results are as Figure 1 shown, Figure 1 a is CeO2-MoS2@PTX@PDLi, Figure 1b is the CeO2-MoS2@PTX biomimetic nanoparticles. The prepared nanoparticles CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles are spherical and have relatively uniform nanoparticle sizes. The diameter of the CeO2-MoS2@PTX biomimetic nanoparticles is increased by about 20-50 nm compared to CeO2-MoS2@PTX@PD-L1i, and a membrane-wrapped state can be seen on its surface, indicating that the 4T1 cancer cell membrane has successfully wrapped on the surface of CeO2-MoS2@PTX@PD-L1i.

[0090] In this invention, in vitro homologous targeting detection was performed on the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1. 4T1 cells and HUMSC cells were respectively seeded in D35-20-1-N dishes, with an initial density of 3×10 5 cells / dish. After the cells adhered, the cells were treated with 1.5 mL of MEM (containing 10% FBS) containing 5 μg / mL of CeO2-MoS2@PTX@PDLi or CeO2-MoS2@PTX biomimetic nanoparticles for 8 h respectively. After the treatment, the cells were washed three times with PBS to remove the free materials, and fixed with 4% paraformaldehyde at room temperature for 20 min. Before imaging, the nuclei were stained with Hoechst for 8 min, washed with PBS, fresh PBS was added, and observed by CLSM. Finally, Image J was used for quantitative analysis of cell uptake. The CLSM observation results are shown in Figure 2 a. Among them, CPPN represents the test group using CeO2-MoS2@PTX biomimetic nanoparticles. After incubating 4T1 cells with CeO2-MoS2@PTX biomimetic nanoparticles for 8 h, the fluorescence signal in the cells is the strongest. The quantitative results of the fluorescence signal in the cells by Image J software are shown in Figure 2 b. The uptake amount of 4T1 cells for CeO2-MoS2@PTX biomimetic nanoparticles is 5 times that of CeO2-MoS2@PTX@PD-L1i, while the uptake amount of HUMSC for CeO2-MoS2@PTX biomimetic nanoparticles is about 2 times that of CeO2-MoS2@PTX@PD-L1i. This result indicates that the CeO2-MoS2@PTX biomimetic nanoparticles modified with 4T1 cancer cell membrane mainly cause an increase in the uptake of tumor cells, and the effect on non-tumor cells is not obvious. Due to the existence of some specific recognition and adhesion molecules on the surface of cancer cells, the nanoparticles modified with cancer cell membrane often have the ability to target tumor cells homologously, and this characteristic helps to improve the targeted delivery of drugs to tumors.

[0091] The present invention detects the in vitro anti-tumor activity of the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1. 4T1 cells were seeded in a 96-well plate at a density of 5000 cells / well. The CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles were used as the experimental groups, and a blank control group was set up. Each group was diluted to a specified concentration gradient with RPMI-1640 medium and administered to the cells. After 48 h of culture, the preparations of each group were aspirated, 20 μL of MTT solution with a concentration of 5 mg / mL was added to each well and incubated for 4 h, and then 100 μL of dimethyl sulfoxide was added to each well and incubated on a constant temperature shaker at 37 °C for 30 min. Finally, the absorbance was measured at a wavelength of 490 nm. The results are as Figure 3 shown. Among them, CPPN represents the test group using the CeO2-MoS2@PTX biomimetic nanoparticles. At a concentration of 10 ng / mL, the lethality rate of the CeO2-MoS2@PTX biomimetic nanoparticles against 4T1 tumor cells is close to 50%, and it increases with the increase in concentration. In addition, compared with CeO2-MoS2@PTX@PD-L1i, its ability to damage tumor cells is more obvious, indicating that the precise damage of the drug is enhanced under the modification of the 4T1 cell membrane, and the combination of paclitaxel and the PD-L1 inhibitor further enhances the killing effect on breast cancer tumor cells.

[0092] The present invention detects the in vitro anti-tumor activity of the biomimetic nanoparticles based on cancer cell membrane modification prepared in Examples 1-5 and Comparative Examples 1-2. 4T1 cells were seeded in a 96-well plate at a density of 5000 cells / well. The CeO2-MoS2@PTX biomimetic nanoparticles prepared in Examples 1-5 and Comparative Examples 1-2 were used as the example groups, and a blank control group was set up. Each group was diluted to 500 ng / L with RPMI-1640 medium and administered to the cells. After 48 h of culture, the preparations of each group were aspirated, 20 μL of MTT solution with a concentration of 5 mg / mL was added to each well and incubated for 4 h, and then 100 μL of dimethyl sulfoxide was added to each well and incubated on a constant temperature shaker at 37 °C for 30 min. Finally, the absorbance was measured at a wavelength of 490 nm. The results are as Figure 4As shown, in the present invention, CeO2 hollow spheres are first prepared, and then CeO2-MoS2 hybrid nanocarriers or CeO2-MoS2 hybrid composite nanocarriers are made from the CeO2 hollow spheres. The CeO2-MoS2 hybrid composite nanocarriers are made by adding polyethylene glycol monoester during the preparation of CeO2-MoS2 hybrid nanocarriers. The polyethylene glycol monoester reacts with hexanoic acid. After adding polyethylene glycol monoester to prepare CeO2-MoS2 hybrid composite nanocarriers, the biomimetic nanoparticles modified with cancer cell membranes prepared therefrom, compared with the biomimetic nanoparticles modified with cancer cell membranes prepared from CeO2-MoS2 hybrid nanocarriers, have a better inhibitory effect on tumor cells, indicating that the structure of polyethylene glycol monoester further helps to exert the effects of PTX and PD-L1i. However, if the amount of polyethylene glycol monoester used is too small or too high, the inhibitory effect of the biomimetic nanoparticles modified with cancer cell membranes on tumor cells will become weak. Keeping the amount of polyethylene glycol monoester used within a certain range helps to improve the inhibitory effect of the biomimetic nanoparticles modified with cancer cell membranes on tumor cells. Further, in the present invention, when preparing CeO2-MoS2@PTX, L-rhamnose bis(ethyl mercaptan) can be added to the PTX solution to prepare CeO2-MoS2@PTX-L, and then the biomimetic nanoparticles modified with cancer cell membranes are further prepared. The resulting product has an improved inhibitory effect on tumor cells, indicating that the use of L-rhamnose bis(ethyl mercaptan) together with PTX and PD-L1i can further improve the inhibitory effect of the biomimetic nanoparticles modified with cancer cell membranes on tumor cells.

[0093] The present invention conducts in vitro cytotoxicity tests on the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1. NCTC1469 cells are seeded in a 96-well plate at a density of 5000 cells / well. The CeO2-MoS2@PTX@PD-L1i and CeO2-MoS2@PTX biomimetic nanoparticles are used as the experimental groups, and a blank control group is set. Each group is diluted to the specified concentration gradient with RPMI-1640 medium and administered to the cells. After culturing for 48 h, the preparations of each group are aspirated. 20 μL of MTT solution with a concentration of 5 mg / mL is added to each well and incubated for 4 h. Then, 100 μL of dimethyl sulfoxide is further added to each well and incubated in a constant temperature shaker at 37 °C for 30 min. Finally, the absorbance is measured at a wavelength of 490 nm. The results are as Figure 5As shown, where CPPN represents the test group using CeO2-MoS2@PTX biomimetic nanoparticles. After MTT detection, it was found that neither CeO2-MoS2@PTX@PD-L1i nor CeO2-MoS2@PTX biomimetic nanoparticles showed obvious cytotoxicity at the detected concentrations. The present invention used the same method to detect the biomimetic nanoparticles prepared in Examples 2-5 based on cancer cell membrane modification, and within the range of 10-10000 ng / mL, there was no obvious cytotoxicity.

[0094] The present invention studied the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1 in breast cancer mice.

[0095] Modeling of breast cancer mice: Resuscitate cryopreserved 4T1 breast cancer cells, culture and passage to an appropriate number, digest with trypsin, centrifuge to obtain cell precipitate, resuspend with PBS, dilute the cell concentration to 5×10 7 cells / mL, place on ice bath, and inoculate 100 μL of cell suspension under the left second pair of mammary fat pads of each mouse. Observe the tumor growth at the inoculation site every day. When the tumor volume reaches about 100 mm 3 or so, it is used for experimental research.

[0096] The calculation formula for tumor volume is: .

[0097] A (unit: mm) is the long diameter of the tumor, and B (unit: mm) is the short diameter of the tumor.

[0098] The present invention tested the in vivo anti-tumor performance of the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1 in breast cancer mice. After the tumor volume of the mice reached about 100 mm 3 , the mice were randomly divided into 5 groups, with 5 mice in each group. The grouping of each group of mice was as follows: PBS group, PBS+L group, CeO2-MoS2@PTX@PDLi group, CeO2-MoS2@PTX biomimetic nanoparticle group, CeO2-MoS2@PTX biomimetic nanoparticle+L group. The dose for each treatment was 5 mg / mL of CeO2-MoS2@PTX biomimetic nanoparticles, and the other groups were equivalently converted. After intravenous injection through the tail vein, light treatment was carried out 24 h after the drug circulated in the mice. The mice in the PBS+L group and the CeO2-MoS2@PTX biomimetic nanoparticle+L group were additionally irradiated with 808 nm (1 W / cm 2)Irradiate the tumor site with light for 5 min. Administer the drug once every two days, and record the changes in body weight and tumor volume every two days for 14 days. Euthanize all mice, remove the subcutaneous tumors of the mice, record the weight of the solid tumors, and perform H&E staining and TUNEL staining on the five internal organs of the mice and the solid tumors. The results are as Figure 6 shown, where CPPN represents the test group using CeO2-MoS2@PTX biomimetic nanoparticles. The volumes of tumors in different test groups at different times are as Figure 6 shown in a. The sizes of tumors in different test groups after 14 days of growth are as Figure 6 shown in b. Compared with other treatments, the CeO2-MoS2@PTX biomimetic nanoparticle + L group has the smallest tumor volume, which is due to the photothermal killing of tumors by near-infrared light irradiation of MoS2, and the combined use of the chemotherapeutic drug PTX and the immunotherapeutic drug PD-L1 inhibitor, the combined action of these three treatment methods. The CeO2-MoS2@PTX biomimetic nanoparticle group has better efficacy than the CeO2-MoS2@PTX@PDLi group, but the former has a more obvious treatment effect, indicating that the nano-drug modified by 4T1 cell membrane can enter the tumor site more and effectively release PTX to exert its efficacy. The CeO2-MoS2@PTX@PDLi group also has a certain tumor inhibitory effect, but the effect is far from that of the 4T1 cell membrane modification group, indicating that CeO2-MoS2@PTX@PDLi has poor anti-cancer effect on this drug-resistant tumor. By performing H&E staining on the main organs such as the heart, liver, spleen, lungs and kidneys of the mice, the results are as Figure 6 shown in c. It can be seen that there is no obvious difference between the CeO2-MoS2@PTX biomimetic nanoparticle group and the PBS group, indicating that the CeO2-MoS2@PTX biomimetic nanoparticles have good biosafety. The results of TUNEL staining are shown as Figure 6 shown in d. In the mice treated with CeO2-MoS2@PTX biomimetic nanoparticles and laser, more green cells, that is, apoptotic cells, were observed. Compared with the single treatment group, apoptosis was more obvious in the CeO2-MoS2@PTX biomimetic nanoparticle + L treatment group. The results prove that this carrier can better kill tumor cells through chemotherapy, immunotherapy and photothermal therapy.

[0099] In this invention, in vivo fluorescence imaging was performed on breast cancer mice using the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1. In vivo fluorescence imaging was mainly due to the loading of the fluorescent dye IR820 on the test samples. Under the irradiation of an 808 nm laser, the fluorescent dye was activated to release optical signals. Nine mice were selected and divided into three groups. The circulation time of each group of drugs in the mice was 4 h, 8 h, 12 h, 24 h, 48 h, and 72 h respectively. In the first group, 100 μL of the CeO2-MoS2@PTX biomimetic nanoparticle solution was injected at different time points. The CeO2-MoS2@PTX biomimetic nanoparticle solution was prepared by dispersing CeO2-MoS2@PTX biomimetic nanoparticles in PBS containing IR820, with a concentration of 5 mg / mL. In the second group, 100 μL of the CeO2-MoS2@PTX@PDLi solution was injected at each time point. The CeO2-MoS2@PTX@PDLi solution was prepared by dispersing CeO2-MoS2@PTX@PDLi in PBS containing IR820, with a concentration of 5 mg / mL. In the third group, 100 μL of the IR820 solution was injected at different time points. The IR820 solution was prepared by dispersing IR820 in PBS. The concentration of IR820 in different test groups was the same, and the concentration of IR820 was approximately 0.67 mg / mL. After anesthetizing the mice intraperitoneally at each time point, the fluorescence signals in the mice were observed by in vivo animal imaging. The in vivo distribution of CeO2-MoS2@PTX biomimetic nanoparticles in Balb / c mice bearing 4T1 tumors was studied by TR820 fluorescence imaging technology. Figure 7 The results showed that, among them, CPPN represents the test group using CeO2-MoS2@PTX biomimetic nanoparticles, CeO2-MoS2@PTX@PDLi represents the test group of CeO2-MoS2@PTX@PDLi, and IR820 represents the test group of IR820. When the nanoparticles were intravenously injected into 4T1 breast cancer-bearing mice, the free IR820 was rapidly cleared from the body, and no obvious signal was detected after 12 h. Compared with free IR820, CeO2-MoS2@PTX@PDLi enhanced the in vivo retention by enhancing the EPR effect of NPs, and a small amount of tumor accumulation was observed at 24 h and 48 h. Throughout all the test times, a large amount of CeO2-MoS2@PTX biomimetic nanoparticles were distributed in the tumor site, and the concentration of CeO2-MoS2@PTX biomimetic nanoparticles in the tumor reached the highest level 24 h after injection, indicating that the CeO2-MoS2@PTX biomimetic nanoparticles exhibited a higher tumor-targeting accumulation ability. It is speculated that the high accumulation of CeO2-MoS2@PTX biomimetic nanoparticles in tumors may be due to the homologous active targeting potential of the cell membrane coating on the nanoparticles and the significantly extended circulation time.

[0100] The above-described embodiments and / or implementation manners are only used to illustrate the preferred embodiments and / or implementation manners for implementing the technology of the present invention, and do not impose any formal restrictions on the implementation manners of the technology of the present invention. Any person skilled in the art, without departing from the scope of the technical means disclosed in the content of the present invention, may make some modifications or changes to other equivalent embodiments, but should still be regarded as the same technology or embodiment as the present invention in essence.

[0101] Specific examples are used in this article to elaborate on the principles and implementation manners of the present application. The description of the above embodiments is only used to help understand the method and its core idea of the present application. The above is only the preferred implementation manner of the present application. It should be noted that due to the limited nature of written expression and the objectively infinite specific structures, for those of ordinary skill in the art, without departing from the principles of the present application, several improvements, refinements or changes can be made, or the above technical features can be combined in an appropriate manner; these improvements, refinements, changes or combinations, or directly applying the concept and technical solution of the invention to other occasions without improvement, should all be regarded as the protection scope of the present application.

Claims

1. A biomimetic nanoparticle modified based on cancer cell membranes, comprising: A carrier, an effector agent loaded on the carrier, and an outer coating of cancer cell membrane; the carrier contains CeO2 and MoS2, CeO2 has a hollow structure, and MoS2 is combined with the hollow structure of CeO2.

2. The biomimetic nanoparticles based on cancer cell membrane modification according to claim 1, wherein: The effector agent includes at least one of PTX and PDL1i, and PDL1i is a PDL1 antibody; or, the cancer cell membrane is a 4T1 cell membrane.

3. A preparation method of a biomimetic nanoparticle modified with a cancer cell membrane, comprising: S1, preparing a carrier by reacting CeO2 hollow spheres with sodium molybdate in a carrier reaction solution; the carrier reaction solution also contains thioacetamide; S2, treating the carrier with an effector agent solution to prepare a carrier-effector agent loaded with the effector agent; the effector agent solution contains the effector agent, and the effector agent includes at least one of PTX and PDL1i, and PDL1i is a PDL1 antibody; S3, mixing the carrier-effector agent with a cancer cell membrane suspension to prepare a biomimetic nanoparticle modified with a cancer cell membrane, and the cancer cell membrane suspension contains a cancer cell membrane.

4. The preparation method of a biomimetic nanoparticle based on cancer cell membrane modification according to claim 3, wherein: The carrier reaction solution is prepared by dispersing CeO2 hollow spheres in deionized water, and then adding sodium molybdate and thioacetamide; the usage amount of CeO2 hollow spheres is 0.05-0.2 wt% of deionized water, the usage amount of sodium molybdate is 200-300 wt% of CeO2 hollow spheres, and the usage amount of thioacetamide is 400-600 wt% of CeO2 hollow spheres; or, The carrier reaction solution is prepared by dispersing CeO2 hollow spheres in deionized water, and then adding sodium molybdate, thioacetamide and polyethylene glycol monoester, and the polyethylene glycol monoester is prepared by reacting polyethylene glycol with fatty acid; the usage amount of CeO2 hollow spheres is 0.05-0.2 wt% of deionized water, the usage amount of sodium molybdate is 200-300 wt% of CeO2 hollow spheres, the usage amount of thioacetamide is 400-600 wt% of CeO2 hollow spheres, and the usage amount of polyethylene glycol monoester is 20-100 wt% of sodium molybdate dihydrate.

5. The preparation method of a biomimetic nanoparticle based on cancer cell membrane modification according to claim 3, characterized in that: The effector agent solution includes a PTX solution and a solution containing PDL1i; in the preparation of the carrier-effector agent, first, the carrier is mixed with the PTX solution to obtain carrier@PTX, and then carrier@PTX is mixed with the solution containing PDL1i to obtain carrier@PTX@PDL1i, that is, the carrier-effector agent.

6. The preparation method of a biomimetic nanoparticle based on cancer cell membrane modification according to claim 5, wherein: The PTX solution is prepared by mixing PTX and deionized water, the PTX solution contains 0.05-0.2 wt% of PTX, the PTX solution is measured by PTX, and the usage amount of the carrier is 100-300 wt% of PTX.

7. The preparation method of a biomimetic nanoparticle based on cancer cell membrane modification according to claim 3, characterized in that: The cancer cell membrane suspension is prepared by resuspending the cancer cell membrane in a PBS solution containing PMSF, and the pH of the PBS solution is 7-7.8; the PBS solution contains 0.5-2 mM of PMSF, and the usage amount relationship between the cancer cell membrane and the PBS solution is 0.1-2 mg / mL.

8. The preparation method of a biomimetic nanoparticle based on cancer cell membrane modification according to claim 3, characterized in that: The usage amount of the cancer cell membrane is 80-120 wt% of the carrier-effector agent.

9. Use of the biomimetic nanoparticle modified with a cancer cell membrane prepared by the preparation method according to any one of claims 3-8 in the preparation of an anti-tumor drug.

10. An anti-tumor drug, comprising the biomimetic nanoparticles modified based on cancer cell membranes according to any one of claims 1-2.

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