Biomimetic nanoparticles modified with cancer cell membranes and their preparation and application

Abstract

The present invention discloses biomimetic nanoparticles modified with cancer cell membranes, their preparation methods, and applications. These nanoparticles, belonging to the field of nano-drug delivery technology, specifically relate to biomimetic nanoparticles modified with cancer cell membranes, comprising: a carrier, an agent loaded on the carrier, and an outer layer of a cancer cell membrane; the carrier contains CeO2 and MoS2, the CeO2 having a hollow structure, and the MoS2 bound to the hollow CeO2 structure. The biomimetic nanoparticles modified with cancer cell membranes prepared by the present invention exhibit excellent targeting, anti-tumor activity, low cytotoxicity, tumor growth inhibition, excellent tumor-targeting accumulation, long circulation time, and can effectively kill tumor cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of nano-drug delivery, and in particular relates to biomimetic nanoparticles based on cancer cell membrane modification, and a preparation method and application thereof. Background Art

[0002] Breast cancer is a common malignancy that poses a serious threat to women's health and life. Treatment options for breast cancer primarily include surgery, radiotherapy, chemotherapy, endocrine therapy, targeted therapy, and immunotherapy, but breast cancer still carries high recurrence and mortality rates. In recent years, research into multi-modality combination therapies has deepened. Chemoimmunotherapy, among others, leverages the cytotoxic and immunomodulatory properties of chemotherapeutic drugs to offer a synergistic anti-tumor treatment approach with broad potential. While chemoimmunotherapy demonstrates significant potential for cancer treatment, a key challenge remains how to enhance the synergistic therapeutic effect while effectively mitigating the side effects of multiple drugs. In recent decades, drug delivery systems have emerged as a cutting-edge strategy to address this challenge. As novel delivery vehicles for chemoimmunotherapy, they can significantly prolong drug action, promote drug accumulation within the lesion, and precisely direct the drug to specific therapeutic targets, minimizing potential harm to non-target tissues. This advancement has opened up a more precise and personalized avenue for chemoimmunotherapy of breast cancer.

[0003] Molybdenum disulfide (MoS2), a layered transition metal sulfide with a large specific surface area, can be modified and loaded with drug molecules or immune adjuvants for targeted delivery and controlled release. Furthermore, MoS2 can generate localized hyperthermia under near-infrared light irradiation, triggering immunogenic cell death (ICD) in tumor cells and enhancing the efficacy of immunotherapy. However, compared with other nanomaterials such as gold nanoparticles, the photothermal conversion efficiency of MoS2 nanosheets is relatively low, which to some extent limits their application in cancer-photothermal therapy. Furthermore, immune clearance, poor antigen presentation in tumor tissue, and the lack of specific targets are currently the greatest obstacles to MoS2 nano-drug delivery systems. Therefore, the development of novel MoS2-based nano-drug delivery and immunotherapy systems is particularly important. Summary of the Invention

[0004] The purpose of the present invention is to provide a biomimetic nanoparticle based on cancer cell membrane modification that has good targeting, good anti-tumor activity, low cytotoxicity, inhibits tumor growth, has good tumor targeting accumulation ability, long circulation time, and can effectively kill tumor cells, as well as its preparation method and application.

[0005] The technical solutions adopted by the present invention to achieve the above-mentioned purpose are:

[0006] A biomimetic nanoparticle modified with a cancer cell membrane comprises: a carrier, an agent loaded on the carrier, and an outer layer coated with a cancer cell membrane; the carrier contains CeO2 and MoS2, with the CeO2 forming a hollow structure and the MoS2 bound to the hollow CeO2 structure. The invention first prepares a hybrid nanostructure containing CeO2 and MoS2 as a carrier through chemical synthesis, then mechanically embeds PTX into the carrier to obtain nanoparticles with controlled drug release. Subsequently, a PD-L1 inhibitor is coupled to the surface of the nanoparticles through an amidation reaction. Finally, extracted 4T1 cancer cell membranes are wrapped around the outer surface of the aforementioned product through mechanical extrusion, resulting in cancer cell membrane-modified biomimetic nanoparticles (CPPNs) capable of targeted drug release. The CPPN nanoparticles provided by the present invention, in which the carrier containing a CeO2 and MoS2 hybrid nanostructure not only provides a drug-carrying framework but also serves as a photothermal agent. PTX, a common chemotherapy drug for breast cancer, is combined with the PD-L1 inhibitor used in immunotherapy for treatment. Coating the nanoparticles with 4T1 cell membranes can enhance the efficacy of the biomimetic nanoparticles prepared in the present invention, which are modified based on cancer cell membranes. This invention provides an application foundation for nanomaterials containing CeO2 and MoS2 hybrid nanostructures as carriers for drug-carrying immunotherapies, and also provides a basis for applying such drug-carrying systems in breast cancer treatment.

[0007] Preferably, the activating agent comprises at least one of PTX and PDL1i, and PDL1i is a PDL1 antibody; or the cancer cell membrane is a 4T1 cell membrane.

[0008] The present invention discloses a method for preparing biomimetic nanoparticles based on cancer cell membrane modification, comprising:

[0009] S1, a carrier is prepared by reacting CeO2 hollow spheres with sodium molybdate in a carrier reaction solution; the carrier reaction solution also contains thioacetamide;

[0010] S2, treating a carrier and an agent solution to prepare a carrier-agent loaded with an agent; the agent solution contains an agent, the agent including at least one of PTX and PDL1i, and PDL1i is a PDL1 antibody;

[0011] S3, a carrier-effective agent is mixed with a cancer cell membrane suspension to prepare biomimetic nanoparticles modified based on the cancer cell membrane, and the cancer cell membrane suspension contains the cancer cell membrane.

[0012] Preferably, the carrier reaction solution is prepared by dispersing CeO2 hollow balls in deionized water, and then adding sodium molybdate and thioacetamide; the amount of CeO2 hollow balls used is 0.05-0.2wt% of deionized water, the amount of sodium molybdate used is 200-300wt% of CeO2 hollow balls, and the amount of thioacetamide used is 400-600wt% of CeO2 hollow balls; or,

[0013] The carrier reaction liquid is prepared by dispersing CeO2 hollow balls in deionized water, and then adding sodium molybdate, thioacetamide and polyethylene glycol monoester, wherein the polyethylene glycol monoester is prepared by reacting polyethylene glycol with fatty acids; the amount of CeO2 hollow balls used is 0.05-0.2wt% of deionized water, the amount of sodium molybdate used is 200-300wt% of the CeO2 hollow balls, the amount of thioacetamide used is 400-600wt% of the CeO2 hollow balls, and the amount of polyethylene glycol monoester used is 20-100wt% of sodium molybdate dihydrate. The present invention introduces a polyethylene glycol monoester prepared by reacting polyethylene glycol with fatty acids during the process of combining MoS2 with CeO2 hollow spheres, so that the resulting carrier has a polyethylene glycol monoester structure. Through the structure of the polyethylene glycol monoester and the structures of MoS2 and CeO2, and under the action of the framework of CeO2 and MoS2 and a photothermal agent, the introduction of the polyethylene glycol monoester structure can better exert the effect of the drug and improve the therapeutic effect on tumors. The amount of polyethylene glycol monoester used should not be too small, as too little will not be effective. At the same time, too much polyethylene glycol monoester used will hinder the effect of drug treatment. Therefore, the amount of polyethylene glycol monoester used needs to be within a certain range to achieve a good effect.

[0014] Preferably, the fatty acid is a C3-12 saturated hydrocarbon acid or an unsaturated hydrocarbon acid.

[0015] More preferably, the fatty acid is hexanoic acid.

[0016] Preferably, the efficacious agent solution includes a PTX solution and a PDL1i-containing solution; in the preparation of the carrier-efficacious agent, the carrier is first mixed with the PTX solution to obtain carrier@PTX, and then the carrier@PTX is mixed with the PDL1i-containing solution to obtain carrier@PTX@PDL1i, that is, carrier-efficacious agent.

[0017] 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 based on PTX, and the amount of the carrier used is 100-300 wt % of PTX.

[0018] Preferably, the cancer cell membrane suspension is prepared by resuspending the cancer cell membrane in a PBS solution containing PMSF, the pH of the PBS solution is 7-7.8; the PBS solution contains 0.5-2 mM PMSF, and the usage ratio of the cancer cell membrane to the PBS solution is 0.1-2 mg / mL.

[0019] Preferably, the amount of cancer cell membrane used is 80-120 wt % of the carrier-effective agent.

[0020] The present invention discloses the use of bionic nanoparticles based on cancer cell membrane modification prepared by the above preparation method in the preparation of anti-tumor drugs.

[0021] Preferably, in the preparation of CeO2 hollow spheres, ammonium hydroxide solution is added to a tetraethyl silicate mixture, stirred at 20-40°C for 12-48h, centrifuged and dried to obtain SiO2, and then SiO2 is dispersed in ethylene glycol, cerium nitrate hexahydrate and deionized water are added, stirred and mixed evenly, transferred to a reactor, and heat treated at 120-160°C for 12-48h, washed and dried after centrifugation to obtain SiO2@CeO2 particles, and finally dispersed in an alkaline solution for 1-4d, washed and dried after centrifugation to obtain CeO2 hollow spheres.

[0022] More preferably, in the preparation of CeO2 hollow spheres, the ammonium hydroxide solution is prepared by mixing ammonium hydroxide and deionized water, and the ammonium hydroxide solution contains 6-18 wt% of ammonium hydroxide.

[0023] More preferably, in the preparation of CeO2 hollow spheres, the tetraethyl silicate mixed solution is prepared by mixing tetraethyl silicate and ethanol, and the tetraethyl silicate mixed solution contains 1-5 wt% of tetraethyl silicate.

[0024] More preferably, in the preparation of CeO2 hollow spheres, when the ammonium hydroxide solution is mixed with the tetraethyl silicate mixture, the ammonium hydroxide solution is measured as ammonium hydroxide, the tetraethyl silicate mixture is measured as tetraethyl silicate, and the amount of ammonium hydroxide used is 80-120wt% of the tetraethyl silicate.

[0025] More preferably, in the preparation of CeO2 hollow spheres, the amount of SiO2 used is 0.3-2.4 wt% of ethylene glycol.

[0026] More preferably, in the preparation of CeO2 hollow spheres, the amount of cerium nitrate hexahydrate used is 400-600 wt% of SiO2.

[0027] More preferably, in the preparation of CeO2 hollow spheres, the amount of deionized water used is 700-800wt% of SiO2, the alkaline solution is a sodium hydroxide solution, and the concentration of sodium hydroxide in the sodium hydroxide solution is 2-6M.

[0028] Preferably, in the preparation of CeO2-MoS2 hybrid nanocarriers, CeO2 hollow spheres are dispersed in deionized water, ultrasonically treated for 10-60 minutes, then sodium molybdate and thioacetamide are added, stirred for 10-60 minutes, then transferred to a reactor, treated at 190-220°C for 12-48 hours, centrifuged, washed and dried to obtain CeO2-MoS2 hybrid nanocarriers.

[0029] More preferably, in the preparation of CeO2-MoS2 hybrid nanocarriers, the amount of CeO2 hollow spheres used is 0.05-0.2 wt% of deionized water.

[0030] More preferably, in the preparation of CeO2-MoS2 hybrid nanocarriers, the amount of sodium molybdate used is 200-300 wt% of the CeO2 hollow spheres.

[0031] More preferably, in the preparation of CeO2-MoS2 hybrid nanocarriers, the amount of thioacetamide used is 400-600 wt% of the CeO2 hollow spheres.

[0032] Preferably, in the preparation of polyethylene glycol monoester, polyethylene glycol and fatty acid are added to xylene, and then a catalyst is added, and the mixture is reacted at 150-170° C. for 3-12 hours. After the reaction is completed, the mixture is washed with toluene, isopropanol and saturated brine in sequence, and vacuum dried to obtain polyethylene glycol monoester.

[0033] More preferably, in the preparation of polyethylene glycol monoester, the molecular weight of polyethylene glycol is 100-500, and the amount of polyethylene glycol used is 10-50 wt % of xylene.

[0034] More preferably, in the preparation of polyethylene glycol monoester, the fatty acid is hexanoic acid, and the molar amount of the fatty acid used is 60-100% of the molar amount of the polyethylene glycol used.

[0035] More preferably, in the preparation of polyethylene glycol monoester, the catalyst is a zirconium n-butoxide solution, which is prepared by mixing zirconium n-butoxide and isopropanol, the zirconium n-butoxide solution has a zirconium n-butoxide content of 0.05-0.3wt%, and the amount of the catalyst used is 0.5-3wt% of the polyethylene glycol.

[0036] Preferably, in the preparation of CeO2-MoS2 hybrid composite nanocarriers, CeO2 hollow spheres are dispersed in deionized water, ultrasonically treated for 10-60 minutes, then sodium molybdate and thioacetamide are added, stirred for 10-60 minutes, then polyethylene glycol monoester is added and mixed, and then transferred to a reactor, treated at 190-220°C for 12-48 hours, centrifuged, washed and dried to obtain CeO2-MoS2 hybrid nanocarriers.

[0037] More preferably, in the preparation of CeO2-MoS2 hybrid composite nanocarrier, the amount of CeO2 hollow spheres used is 0.05-0.2 wt% of deionized water.

[0038] More preferably, in the preparation of CeO2-MoS2 hybrid composite nanocarriers, the amount of sodium molybdate used is 200-300 wt% of the CeO2 hollow spheres.

[0039] More preferably, in the preparation of CeO2-MoS2 hybrid composite nanocarriers, the amount of thioacetamide used is 400-600 wt% of the CeO2 hollow spheres.

[0040] More preferably, in the preparation of CeO2-MoS2 hybrid composite nanocarrier, the amount of polyethylene glycol monoester used is 20-100 wt % of sodium molybdate dihydrate.

[0041] Preferably, in the preparation of CeO2-MoS2@PTX, the CeO2-MoS2 hybrid nanocarrier is added to the PTX solution, stirred for 10-48 hours under light-proof conditions, washed after centrifugation, and freeze-dried to obtain CeO2-MoS2@PTX.

[0042] More preferably, in the preparation of CeO2-MoS2@PTX, the PTX solution is prepared by mixing PTX and deionized water, and the PTX solution contains 0.05-0.2 wt% of PTX.

[0043] More preferably, in the preparation of CeO2-MoS2@PTX, the PTX solution is measured as PTX, and the amount of CeO2-MoS2 hybrid nanocarrier used is 100-300 wt% of PTX.

[0044] Preferably, in the preparation of CeO2-MoS2@PTX, CeO2-MoS2 hybrid nanocarrier and L-rhamnosyl diethyl mercaptan are added to a PTX solution, stirred for 10-48 hours under dark conditions, washed after centrifugation, and freeze-dried to obtain CeO2-MoS2@PTX. When loading PTX, the present invention can also add L-rhamnosyl diethyl mercaptan to the drug, and L-rhamnosyl diethyl mercaptan and PTX are jointly loaded in the CeO2-MoS2 hybrid nanocarrier, so that CeO2-MoS2@PTX contains L-rhamnosyl diethyl mercaptan in addition to PTX. The use of L-rhamnosyl diethyl mercaptan can improve the therapeutic effect of the biomimetic nanoparticles based on the cancer cell membrane on tumors under the action of other components of the biomimetic nanoparticles based on the cancer cell membrane.

[0045] More preferably, in the preparation of CeO2-MoS2@PTX, the PTX solution is prepared by mixing PTX and deionized water, and the PTX solution contains 0.05-0.2 wt% of PTX.

[0046] More preferably, in the preparation of CeO2-MoS2@PTX, the PTX solution is measured as PTX, and the amount of CeO2-MoS2 hybrid nanocarrier used is 100-300 wt% of PTX.

[0047] More preferably, in the preparation of CeO2-MoS2@PTX, the amount of L-rhamnosyl diethylmercaptan used is 3-21 wt % of PTX.

[0048] Preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, CeO2-MoS2@PTX, EDC·HCl and NHS are added to a PBS solution, ultrasonically treated for 10-60 minutes, and then PDL1 antibody is added, and the mixture is stirred in the dark at 20-40°C for 10-48 hours to obtain CeO2-MoS2@PTX@PDL1i.

[0049] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the pH of the PBS solution is 7-7.8.

[0050] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the amount of CeO2-MoS2@PTX used is 0.05-0.4 wt% of the PBS solution.

[0051] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the amount of EDC·HCl used is 100-300 wt% of CeO2-MoS2@PTX.

[0052] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the amount of NHS used is 100-300 wt% of CeO2-MoS2@PTX.

[0053] More preferably, in the preparation of CeO2-MoS2@PTX@PDL1i, the amount of PDL1 antibody used is 1-10 wt % of CeO2-MoS2@PTX.

[0054] Preferably, in the extraction of cancer cell membranes, 4T1 cells are cultured in DMEM medium. After the 4T1 cells are fully grown, the 4T1 cells are removed, resuspended in PBS solution, centrifuged to collect the precipitate, resuspended in hypotonic lysis buffer, and treated in an ice bath for 5-30 minutes. The precipitate is then repeatedly frozen and thawed 2-5 times in liquid nitrogen and a 30-40°C water bath, centrifuged at 500-1000g for 5-15 minutes at 0-10°C, the supernatant is collected, and the supernatant is centrifuged again for 10-60 minutes. The precipitate is collected, resuspended in hypotonic lysis buffer, and centrifuged again at 10000-18000g for 10-60 minutes at 0-10°C to obtain cancer cell membranes.

[0055] More preferably, during the extraction of cancer cell membranes, the DMEM culture medium contains 5-15% FBS, and the DMEM culture medium further contains penicillin and streptomycin, with the amount of penicillin being 20-100 IU / mL and the amount of streptomycin being 20-100 IU / mL.

[0056] More preferably, during the extraction of cancer cell membranes, the pH of the PBS solution is 7-7.8.

[0057] 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.

[0058] More preferably, in the extraction of cancer cell membranes and the resuspension process, the amount of solid matter and liquid reagent used in the resuspension is 0.1-1 mg / mL.

[0059] Preferably, in the preparation of CeO2-MoS2@PTX biomimetic nanoparticles, the cancer cell membrane is added to a PBS solution containing PMSF and resuspended, and then CeO2-MoS2@PTX@PDL1i is added and mixed, and then mechanically extruded after being coated with a polycarbonate film, and then ultrasonicated in a water bath at 0-10°C in the dark for 10-60 min to obtain CeO2-MoS2@PTX biomimetic nanoparticles.

[0060] More preferably, in the preparation of CeO2-MoS2@PTX biomimetic nanoparticles, the pH of the PBS solution is 7-7.8.

[0061] More preferably, in the preparation of CeO2-MoS2@PTX biomimetic nanoparticles, the PBS solution contains 0.5-2 mM PMSF, and the usage ratio of the cancer cell membrane to the PBS solution is 0.1-2 mg / mL.

[0062] More preferably, in the preparation of CeO2-MoS2@PTX biomimetic nanoparticles, the amount of cancer cell membrane used is 80-120 wt % of CeO2-MoS2@PTX@PDL1i.

[0063] The present invention discloses an anti-tumor drug, comprising the above-mentioned bionic nanoparticles based on cancer cell membrane modification.

[0064] The present invention utilizes biomimetic nanoparticles modified based on cancer cell membranes, which are formed by a carrier, an agent loaded on the carrier, and an outer layer of cancer cell membranes. The carrier is composed of CeO2 and MoS2, wherein the CeO2 is a hollow structure, and then MoS2 is bonded to the carrier through synthesis, and then further loaded with agents including PTX and PDL1i, and finally coated with the cancer cell membranes. When preparing the carrier, polyethylene glycol monoester can be added when preparing the carrier bonded with MoS2 from the hollow CeO2. The polyethylene glycol monoester is prepared by reacting polyethylene glycol with fatty acids. Therefore, the biomimetic nanoparticles modified based on cancer cell membranes prepared by the present invention have good targeting, good anti-tumor activity, low cytotoxicity, tumor growth inhibition, tumor-targeted accumulation ability, long circulation time, and can effectively kill tumor cells. Therefore, the present invention is a biomimetic nanoparticle modified based on cancer cell membranes, which has good targeting, good anti-tumor activity, low cytotoxicity, tumor growth inhibition, tumor-targeted accumulation ability, long circulation time, and can effectively kill tumor cells, as well as a preparation method and application thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 This is an electron microscope image of biomimetic nanoparticles.

[0066] Figure 2 This is the result of in vitro homologous targeting detection.

[0067] Figure 3 This is a graph showing the results of in vitro anti-tumor activity testing.

[0068] Figure 4 This figure shows the results of in vitro anti-tumor activity testing of biomimetic nanoparticles modified with cancer cell membranes.

[0069] Figure 5 The figure shows the results of in vitro cytotoxicity test.

[0070] Figure 6 This is a diagram showing the results of in vivo anti-tumor performance testing.

[0071] Figure 7 This is an in vivo fluorescence imaging image. DETAILED DESCRIPTION

[0072] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. 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.

[0073] The following first describes the concepts involved in this application with reference to the accompanying drawings. It should be noted that the following description of each concept is intended only to make the content of this application easier to understand and does not limit the scope of protection of this application. At the same time, the embodiments and features in the embodiments of this application can be combined with each other unless there is a conflict. The following detailed description of this application will be made with reference to the accompanying drawings and in conjunction with the embodiments.

[0074] Example 1: A method for preparing biomimetic nanoparticles based on cancer cell membrane modification

[0075] Preparation of CeO2 hollow spheres: Add ammonium hydroxide solution to a tetraethyl silicate mixture, stir at 25°C for 24 hours, centrifuge 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 reactor, and heat treat at 130°C for 24 hours, wash and dry after centrifugation to obtain SiO2@CeO2 particles, finally disperse in an alkaline solution for 2 days, wash and dry after centrifugation to obtain CeO2 hollow spheres. The ammonium hydroxide solution is formed by mixing ammonium hydroxide and deionized water, and the ammonium hydroxide solution contains 13wt% of ammonium hydroxide. The tetraethyl silicate mixed solution is formed by mixing tetraethyl silicate and ethanol, and the tetraethyl silicate mixed solution contains 2.78wt% of tetraethyl silicate. When the ammonium hydroxide solution and the tetraethyl silicate mixed solution are mixed, the ammonium hydroxide solution is measured with ammonium hydroxide, and the tetraethyl silicate mixed solution is measured with tetraethyl silicate. The amount of ammonium hydroxide used is 105wt% of the tetraethyl silicate; the amount of SiO2 used is 0.77wt% of ethylene glycol, the amount of cerium nitrate hexahydrate used is 500wt% of SiO2, the amount of deionized water used is 750wt% of SiO2, and the alkaline solution is a sodium hydroxide solution, and the concentration of sodium hydroxide in the sodium hydroxide solution is 5M.

[0076] Preparation of CeO2-MoS2 hybrid nanocarriers: CeO2 hollow spheres were dispersed in deionized water and ultrasonically treated for 30 minutes. Sodium molybdate and thioacetamide were then added and stirred for 30 minutes. The mixture was then transferred to a reactor and treated at 200°C for 24 hours. After centrifugation, the mixture was washed and dried to obtain CeO2-MoS2 hybrid nanocarriers. The CeO2 hollow spheres were used in an amount of 0.14 wt% of the deionized water, 250 wt% of the sodium molybdate, and 500 wt% of the thioacetamide.

[0077] Preparation of CeO2-MoS2@PTX: CeO2-MoS2 hybrid nanocarriers were added to a PTX solution, stirred in the dark for 24 hours, centrifuged, washed, and freeze-dried to obtain CeO2-MoS2@PTX. The PTX solution was prepared by mixing PTX and deionized water. The PTX solution contained 0.1 wt% PTX. The PTX solution was measured based on PTX, and the amount of CeO2-MoS2 hybrid nanocarriers used was 200 wt% of the PTX.

[0078] Preparation of CeO2-MoS2@PTX@PDL1i: CeO2-MoS2@PTX, EDC·HCl, and NHS were added to a PBS solution and sonicated for 20 minutes. PDL1 antibody was then added and stirred at 25°C in the dark for 24 hours to obtain CeO2-MoS2@PTX@PDL1i. The pH of the PBS solution was 7.4. The amount of CeO2-MoS2@PTX was 0.3 wt% of the PBS solution, the amount of EDC·HCl was 200 wt% of the CeO2-MoS2@PTX, the amount of NHS was 200 wt% of the CeO2-MoS2@PTX, and the amount of PDL1 antibody was 5 wt% of the CeO2-MoS2@PTX.

[0079] Cancer cell membrane extraction: 4T1 cells were cultured in DMEM medium. Once confluent, the cells were removed and resuspended in PBS. The pellet was collected by centrifugation and resuspended in hypotonic lysis buffer. The pellet was incubated on ice for 100 min. The pellet was then freeze-thawed three times in liquid nitrogen and a 37°C water bath. The pellet was centrifuged at 700 g for 10 min at 4°C, and the supernatant was collected. The pellet was centrifuged again for 30 min, and the pellet was resuspended in hypotonic lysis buffer. The cell membranes were then centrifuged again at 14,000 g for 30 min at 4°C. DMEM medium contained 10% FBS and 50 IU / mL penicillin and 50 IU / mL streptomycin. The pH of the PBS solution was 7.4. The hypotonic lysis buffer consisted of PBS containing 1 mM PMSF. The amount of solid matter and liquid reagents used in the resuspension was 0.5 mg / mL.

[0080] Preparation of CeO2-MoS2@PTX biomimetic nanoparticles: Cancer cell membranes were resuspended in a PBS solution containing PMSF. CeO2-MoS2@PTX@PDL1i was then added and mixed. The mixture was coated with a polycarbonate membrane and mechanically extruded. The mixture was then sonicated in a water bath at 4°C in the dark for 20 minutes to obtain CeO2-MoS2@PTX biomimetic nanoparticles. The PBS solution had a pH of 7.4 and contained 1 mM PMSF. The ratio of cancer cell membrane to PBS solution was 0.5 mg / mL, and the amount of cancer cell membrane to PBS solution was 100 wt% of the CeO2-MoS2@PTX@PDL1i.

[0081] Example 2: A method for preparing biomimetic nanoparticles modified with cancer cell membranes

[0082] Compared with Example 1, this embodiment is different in that the CeO2-MoS2 hybrid nanocarrier is replaced by a CeO2-MoS2 hybrid composite nanocarrier in the preparation of CeO2-MoS2@PTX.

[0083] Preparation of polyethylene glycol monoester: Polyethylene glycol and fatty acid are added to xylene, followed by a catalyst. The mixture is reacted at 160°C for 6 hours. After completion of the reaction, the mixture is washed sequentially with toluene, isopropanol, and saturated brine, and then dried under vacuum to obtain the polyethylene glycol monoester. The molecular weight of the polyethylene glycol is 300, and the amount of polyethylene glycol used is 30% by weight of the xylene. The fatty acid is hexanoic acid, and the molar amount of the fatty acid used is 80% of the molar amount of the polyethylene glycol used. The catalyst is a zirconium n-butoxide solution, which is a mixture of zirconium n-butoxide and isopropanol, with a zirconium n-butoxide content of 0.15% by weight. The amount of catalyst used is 1% by weight of the polyethylene glycol.

[0084] Preparation of CeO2-MoS2 hybrid composite nanocarriers: CeO2 hollow spheres were dispersed in deionized water and ultrasonically treated for 30 minutes. Then, sodium molybdate and thioacetamide were added and stirred for 30 minutes. Polyethylene glycol monoester was then added and mixed. The mixture was then transferred to a reactor and treated at 200°C for 24 hours. After centrifugation, it was washed and dried to obtain CeO2-MoS2 hybrid nanocarriers. The amount of CeO2 hollow spheres used was 0.14wt% of the deionized water, the amount of sodium molybdate used was 250wt% of the CeO2 hollow spheres, the amount of thioacetamide used was 500wt% of the CeO2 hollow spheres, and the amount of polyethylene glycol monoester used was 40wt% of the sodium molybdate dihydrate.

[0085] Example 3: A method for preparing biomimetic nanoparticles based on cancer cell membrane modification

[0086] Compared with Example 2, this example differs in the preparation of CeO2-MoS2 hybrid composite nanocarrier.

[0087] Preparation of CeO2-MoS2 hybrid composite nanocarriers: CeO2 hollow spheres were dispersed in deionized water and ultrasonically treated for 30 minutes. Then, sodium molybdate and thioacetamide were added and stirred for 30 minutes. Polyethylene glycol monoester was then added and mixed. The mixture was then transferred to a reactor and treated at 200°C for 24 hours. After centrifugation, it was washed and dried to obtain CeO2-MoS2 hybrid nanocarriers. The amount of CeO2 hollow spheres used was 0.14wt% of the deionized water, the amount of sodium molybdate used was 250wt% of the CeO2 hollow spheres, the amount of thioacetamide used was 500wt% of the CeO2 hollow spheres, and the amount of polyethylene glycol monoester used was 90wt% of the sodium molybdate dihydrate.

[0088] Example 4: A method for preparing biomimetic nanoparticles modified with cancer cell membranes

[0089] Compared with Example 2, this example is different in that CeO2-MoS2@PTX.

[0090] Preparation of CeO2-MoS2@PTX: CeO2-MoS2 hybrid nanocarriers and L-rhamnosyl diethylenethiol were added to a PTX solution, stirred in the dark for 24 hours, centrifuged, washed, and freeze-dried to obtain CeO2-MoS2@PTX. The PTX solution was prepared by mixing PTX and deionized water. The PTX solution contained 0.1 wt% PTX. The PTX solution was measured based on PTX. The amount of CeO2-MoS2 hybrid nanocarrier used was 200 wt% of PTX, and the amount of L-rhamnosyl diethylenethiol used was 5 wt% of PTX.

[0091] Example 5: A method for preparing biomimetic nanoparticles modified with cancer cell membranes

[0092] Compared with Example 2, this example is different in that in the preparation of CeO2-MoS2@PTX@PDL1i, CeO2-MoS2@PTX is replaced by CeO2-MoS2@PTX-L.

[0093] Preparation of CeO2-MoS2@PTX-L: CeO2-MoS2 hybrid nanocarriers and L-rhamnosyl diethylenethiol are added to a PTX solution, stirred in the dark for 10-48 hours, centrifuged, washed, and freeze-dried to obtain CeO2-MoS2@PTX. The PTX solution is prepared by mixing PTX and deionized water. The PTX solution contains 0.05-0.2wt% PTX. The PTX solution is measured based on PTX. The amount of CeO2-MoS2 hybrid nanocarrier used is 100-300wt% of PTX, and the amount of L-rhamnosyl diethylenethiol used is 15wt% of PTX.

[0094] Comparative Example 1: A method for preparing biomimetic nanoparticles based on cancer cell membrane modification

[0095] Compared with Example 2, this comparative example differs in the preparation of the CeO2-MoS2 hybrid composite nanocarrier.

[0096] Preparation of CeO2-MoS2 hybrid composite nanocarriers: CeO2 hollow spheres were dispersed in deionized water and ultrasonically treated for 30 minutes. Then, sodium molybdate and thioacetamide were added and stirred for 30 minutes. Polyethylene glycol monoester was then added and mixed. The mixture was then transferred to a reactor and treated at 200°C for 24 hours. After centrifugation, it was washed and dried to obtain CeO2-MoS2 hybrid nanocarriers. The amount of CeO2 hollow spheres used was 0.14wt% of the deionized water, the amount of sodium molybdate used was 250wt% of the CeO2 hollow spheres, the amount of thioacetamide used was 500wt% of the CeO2 hollow spheres, and the amount of polyethylene glycol monoester used was 10wt% of the sodium molybdate dihydrate.

[0097] Comparative Example 2: A method for preparing biomimetic nanoparticles based on cancer cell membrane modification

[0098] Compared with Example 2, this comparative example differs in the preparation of the CeO2-MoS2 hybrid composite nanocarrier.

[0099] Preparation of CeO2-MoS2 hybrid composite nanocarriers: CeO2 hollow spheres were dispersed in deionized water and ultrasonically treated for 30 minutes. Then, sodium molybdate and thioacetamide were added and stirred for 30 minutes. Polyethylene glycol monoester was then added and mixed. The mixture was then transferred to a reactor and treated at 200°C for 24 hours. After centrifugation, it was washed and dried to obtain CeO2-MoS2 hybrid nanocarriers. The amount of CeO2 hollow spheres used was 0.14wt% of the deionized water, the amount of sodium molybdate used was 250wt% of the CeO2 hollow spheres, the amount of thioacetamide used was 500wt% of the CeO2 hollow spheres, and the amount of polyethylene glycol monoester used was 120wt% of the sodium molybdate dihydrate.

[0100] Test example:

[0101] The present invention uses CeO2-MoS2@PTX@PD-L1i and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1 as representatives for research and testing.

[0102] The present invention uses scanning electron microscopy to characterize the morphology of CeO2-MoS2@PTX@PD-L1i and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1. The results are as follows: Figure 1 As shown, Figure 1 a is CeO2-MoS2@PTX@PDLi, Figure 1 b is CeO2-MoS2@PTX biomimetic nanoparticles. The nanoparticles CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in the present invention are spherical and have relatively uniform nanoparticle size. The diameter of CeO2-MoS2@PTX biomimetic nanoparticles is about 20~50 nm larger than that of CeO2-MoS2@PTX@PD-L1i. The surface of the CeO2-MoS2@PTX biomimetic nanoparticles can be seen to be wrapped with a membrane, indicating that the 4T1 cancer cell membrane is successfully wrapped on the surface of CeO2-MoS2@PTX@PD-L1i.

[0103] The present invention conducted in vitro homologous targeting detection on CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1. 4T1 cells and HUMSC cells were seeded in D35-20-1-N dishes, respectively, with an initial density of 3×10 5 cells / dish. After the cells adhered, they 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. After treatment, the cells were washed three times with PBS to remove 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, and fresh PBS was added, and observed by CLSM. Finally, Image J was used to quantitatively analyze the cell uptake. The CLSM observation results are shown in Figure 2. Figure 2 a, where CPPN represents the test group using CeO2-MoS2@PTX biomimetic nanoparticles. After 4T1 cells were incubated with CeO2-MoS2@PTX biomimetic nanoparticles for 8 hours, the fluorescence signal in the cells was the strongest. The quantitative results of the fluorescence signal in the cells using Image J software are shown in Figure 2. Figure 2 As shown in Figure b, the uptake of CeO2-MoS2@PTX biomimetic nanoparticles by 4T1 cells was five times that of CeO2-MoS2@PTX@PD-L1i, while the uptake of CeO2-MoS2@PTX biomimetic nanoparticles by HUMSCs was approximately twice that of CeO2-MoS2@PTX@PD-L1i. This result indicates that 4T1 cancer cell membrane-modified CeO2-MoS2@PTX biomimetic nanoparticles primarily increased their uptake by tumor cells, with no significant effect on non-tumor cells. Due to the presence of specific recognition and adhesion molecules on the surface of cancer cells, cancer cell membrane-modified nanoparticles often have the ability to homologously target tumor cells, a property that helps improve targeted drug delivery to tumors.

[0104] The present invention tested the in vitro anti-tumor activity of CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1, and 4T1 cells were seeded in 96-well plates at a density of 5000 cells / well. CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles were used as experimental groups, and a blank control group was set up. Each group was diluted to a specified concentration gradient with RPMI-1640 culture medium and administered to the cells. After culturing for 48 hours, each group of preparations was aspirated, 20 μL of 5 mg / mL MTT solution was added to each well and incubated for 4 hours, and then 100 μL of dimethyl sulfoxide was added to each well and incubated at 37°C in a constant temperature shaker for 30 minutes. Finally, the absorbance was measured at a wavelength of 490 nm. The results are as follows Figure 3 As shown in the figure, CPPN represents the test group using CeO2-MoS2@PTX biomimetic nanoparticles. At a concentration of 10 ng / mL, the lethality of CeO2-MoS2@PTX biomimetic nanoparticles for 4T1 tumor cells is close to 50%, and increases with increasing concentration. In addition, compared with CeO2-MoS2@PTX@PD-L1i, its ability to damage tumor cells is more obvious, indicating that the modification of the 4T1 cell membrane enhances the precise damage of the drug, and that the combination of paclitaxel and PD-L1 inhibitors further enhances the killing power of breast cancer tumor cells.

[0105] The present invention performs in vitro anti-tumor activity detection on the biomimetic nanoparticles based on cancer cell membrane modification prepared in Examples 1-5 and Comparative Examples 1-2, and 4T1 cells are seeded in 96-well plates at a density of 5000 cells / well. The CeO2-MoS2@PTX biomimetic nanoparticles prepared in Examples 1-5 and Comparative Examples 1-2 are used as the example group, and a blank control group is set up. Each group is diluted to 500 ng / L with RPMI-1640 culture medium and administered to the cells. After culturing for 48 hours, each group of preparations is aspirated, 20 μL of 5 mg / mL MTT solution is added to each well and incubated for 4 hours, and then 100 μL of dimethyl sulfoxide is added to each well and incubated at 37°C constant temperature shaker for 30 minutes. Finally, the absorbance is measured at a wavelength of 490 nm. The results are as follows 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 prepared by adding polyethylene glycol monoester in the process of preparing the CeO2-MoS2 hybrid nanocarriers, and the polyethylene glycol monoester and hexanoic acid react to form the CeO2-MoS2 hybrid composite nanocarriers. After the polyethylene glycol monoester is added to prepare the CeO2-MoS2 hybrid composite nanocarriers, the bionic nanoparticles based on cancer cell membrane modification prepared therefrom are better than those prepared by the CeO2-MoS2 hybrid nanocarriers. The bionic nanoparticles modified based on cancer cell membrane obtained, and the bionic nanoparticles modified based on cancer cell membrane prepared by CeO2-MoS2 hybrid composite nanocarriers have better inhibitory effect on tumor cells, indicating that the structure of polyethylene glycol monoester further contributes to the effects of PTX and PD-L1i; while too little or too much polyethylene glycol monoester will lead to a weakened inhibitory effect of bionic nanoparticles modified based on cancer cell membrane on tumor cells. Keeping the usage of polyethylene glycol monoester within a certain range will help improve the inhibitory effect of bionic nanoparticles modified based on cancer cell membrane on tumor cells. Furthermore, when preparing CeO2-MoS2@PTX, the present invention can add L-rhamnose diethyl mercaptan to the PTX solution to prepare CeO2-MoS2@PTX-L, and then further prepare bionic nanoparticles modified based on cancer cell membranes. The resulting product has an improved inhibitory effect on tumor cells, indicating that the use of L-rhamnose diethyl mercaptan together with PTX and PD-L1i can further improve the inhibitory effect of bionic nanoparticles modified based on cancer cell membranes on tumor cells.

[0106] The present invention conducted an in vitro cytotoxicity test on the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1, and NCTC1469 cells were seeded in 96-well plates at a density of 5000 cells / well. CeO2-MoS2@PTX@PD-L1i and CeO2-MoS2@PTX biomimetic nanoparticles were used as experimental groups, and a blank control group was set up. Each group was diluted to a specified concentration gradient with RPMI-1640 culture medium and administered to the cells. After culturing for 48 hours, each group of preparations was aspirated, 20 μL of 5 mg / mL MTT solution was added to each well and incubated for 4 hours, and then 100 μL of dimethyl sulfoxide was added to each well and incubated at 37°C in a constant temperature shaker for 30 minutes. Finally, the absorbance was measured at a wavelength of 490 nm. The results are shown in the figure. Figure 5As shown in the figure, CPPN represents the test group using CeO2-MoS2@PTX biomimetic nanoparticles. MTT assays revealed that neither CeO2-MoS2@PTX@PD-L1i nor CeO2-MoS2@PTX biomimetic nanoparticles had significant cytotoxicity at the tested concentrations. The present invention employed the same method to test the cancer cell membrane-modified biomimetic nanoparticles prepared in Examples 2-5. No significant cytotoxicity was observed within the range of 10-10,000 ng / mL.

[0107] The present invention studied the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1 in breast cancer mice.

[0108] Breast cancer mouse model: 4T1 breast cancer cells were thawed and cultured to the appropriate number, digested with trypsin, centrifuged to obtain cell pellets, resuspended in PBS, and diluted to a cell concentration of 5 × 10 7 100 μL of cell suspension was inoculated into the lower part of the second to last mammary fat pad on the left side of each mouse. The tumor growth at the inoculation site was observed daily. The tumor volume reached 100 mm. 3 When it is about 200, it is used for experimental research.

[0109] The formula for calculating tumor volume is: .

[0110] A (unit: mm) is the long diameter of the tumor, and B (unit: mm) is the short diameter of the tumor.

[0111] The present invention tested the in vivo anti-tumor performance of CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1 in breast cancer mice. 3 Afterwards, the mice were randomly divided into 5 groups, with 5 mice in each group. The mice in each group were divided into PBS group, PBS+L group, CeO2-MoS2@PTX@PDLi group, CeO2-MoS2@PTX biomimetic nanoparticles group, and CeO2-MoS2@PTX biomimetic nanoparticles+L group. The dose of each treatment was 5 mg / mL of CeO2-MoS2@PTX biomimetic nanoparticles. The other groups were converted equally. The drugs were injected through the tail vein. After the drugs circulated in the mice for 24 hours, they were treated with light. The mice in the PBS+L group and the CeO2-MoS2@PTX biomimetic nanoparticles+L group were additionally treated with 808 nm (1 W / cm 2) irradiate the tumor site with light for 5 minutes. Dosage was administered every two days, and changes in body weight and tumor volume were recorded every two days for 14 days. All mice were euthanized, subcutaneous tumors were removed, and solid tumor weights were recorded. H&E staining and TUNEL staining were performed on the five internal organs and solid tumors of the mice. The results are shown in the figure below. Figure 6 As shown in the figure, CPPN represents the test group using CeO2-MoS2@PTX biomimetic nanoparticles. The tumor volumes of different test groups at different times are shown in Figure 6 As shown in a, the sizes of tumors in different test groups after 14 days of growth are as follows: Figure 6 As shown in b, compared with other treatments, the CeO2-MoS2@PTX biomimetic nanoparticles + L group has the smallest tumor volume. This is due to the combined effect of three treatment methods: near-infrared light irradiation of MoS2 to kill the tumor through photothermal treatment, and combined with chemotherapy drug PTX and immunotherapy drug PD-L1 inhibitor. The CeO2-MoS2@PTX biomimetic nanoparticles group is more effective than the CeO2-MoS2@PTX@PDLi group, but the former has a more obvious therapeutic effect, indicating that the nanomedicine modified with 4T1 cell membrane can enter the tumor site more and effectively release PTX to exert its therapeutic effect. 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 modified group, indicating that CeO2-MoS2@PTX@PDLi has a poor anti-cancer effect for this drug-resistant tumor. H&E staining of major organs such as the heart, liver, spleen, lungs and kidneys of mice is shown as follows. Figure 6 c, it can be seen that there is no significant difference between the CeO2-MoS2@PTX biomimetic nanoparticles group and the PBS group, indicating that the CeO2-MoS2@PTX biomimetic nanoparticles have good biosafety. TUNEL staining results show that Figure 6 In the d figure, more green cells, indicating apoptotic cells, were observed in mice treated with CeO2-MoS2@PTX biomimetic nanoparticles and laser. Apoptosis was more pronounced in the CeO2-MoS2@PTX biomimetic nanoparticle + laser treatment group compared to the single-treatment group. These results demonstrate that this carrier can effectively kill tumor cells through chemotherapy, immunotherapy, and photothermal therapy.

[0112] This study used in vivo fluorescence imaging in mice with breast cancer using the CeO2-MoS2@PTX@PDLi and CeO2-MoS2@PTX biomimetic nanoparticles prepared in Example 1. This in vivo fluorescence imaging is primarily due to the loading of the test samples with the fluorescent dye IR820, which is activated by 808nm laser irradiation to release an optical signal. Nine mice were divided into three groups, and the drug circulation time in each group was 4 hours, 8 hours, 12 hours, 24 hours, 48 ​​hours, and 72 hours, respectively. The first group received 100 μL of a CeO2-MoS2@PTX biomimetic nanoparticle solution at different time points. The CeO2-MoS2@PTX biomimetic nanoparticle solution consisted of CeO2-MoS2@PTX biomimetic nanoparticles dispersed in PBS containing IR820 at a concentration of 5 mg / mL. The second group received 100 μL of a CeO2-MoS2@PTX@PDLi solution at different time points. The CeO2-MoS2@PTX@PDLi solution consisted of CeO2-MoS2@PTX@PDLi dispersed in PBS containing IR820 at a concentration of 5 mg / mL. The third group received 100 μL of an IR820 solution at different time points. The IR820 concentration was the same across all test groups, approximately 0.67 mg / mL. Mice were anesthetized intraperitoneally at each time point, and in vivo fluorescence signals were observed using in vivo imaging. The in vivo distribution of CeO2-MoS2@PTX biomimetic nanoparticles in Balb / c mice bearing 4T1 tumors was studied using TR820 fluorescence imaging technology. Figure 7 Results showed that, when the nanoparticles were intravenously injected into mice bearing 4T1 breast cancer, free IR820 was rapidly cleared from the body, with no significant signal detected after 12 hours. Compared to free IR820, CeO2-MoS2@PTX@PDLi exhibited enhanced in vivo retention by enhancing the EPR effect of the NPs, with minimal tumor accumulation observed at 24 and 48 hours. CeO2-MoS2@PTX biomimetic nanoparticles were abundantly distributed at the tumor site at all test times, reaching their highest concentration 24 hours after injection, demonstrating that CeO2-MoS2@PTX biomimetic nanoparticles exhibit enhanced tumor-targeting and accumulation. 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 prolonged circulation time.

[0113] The embodiments and / or implementation methods described above are only used to illustrate the preferred embodiments and / or implementation methods for realizing the technology of the present invention, and do not impose any form of limitation on the implementation methods of the technology of the present invention. Any person skilled in the art may make slight changes or modifications to other equivalent embodiments without departing from the scope of the technical means disclosed in the content of the present invention, but they should still be regarded as technologies or embodiments that are essentially the same as the present invention.

[0114] This article uses specific examples to illustrate the principles and implementation methods of this application. The description of the above embodiments is only used to help understand the method and core ideas of this application. The above is only the preferred implementation method of this application. It should be pointed out that due to the limitations of textual expression, there are objectively infinite specific structures. For ordinary technicians in this technical field, without departing from the principles of this application, they can also make several improvements, modifications or changes, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, should be regarded as the scope of protection of this application.

Claims

1. A method for preparing biomimetic nanoparticles modified with cancer cell membranes, comprising: S1, CeO2 hollow spheres and sodium molybdate react in a carrier reaction solution to prepare a carrier; the carrier reaction solution also contains thioacetamide; S2, a carrier-effective agent loaded with an effective agent is prepared by treating a carrier with an effective agent solution; the effective agent solution contains effective agents, including PTX and PDL1i, where PDL1i is a PDL1 antibody; S3, a carrier-effective agent is mixed with a cancer cell membrane suspension to prepare biomimetic nanoparticles modified based on cancer cell membranes, wherein the cancer cell membrane suspension contains cancer cell membranes; the cancer cell membranes are 4T1 cell membranes; The carrier reaction liquid is prepared by dispersing CeO2 hollow balls in deionized water, and then adding sodium molybdate, thioacetamide, and polyethylene glycol monoester, wherein the polyethylene glycol monoester is prepared by reacting polyethylene glycol with fatty acids; the amount of CeO2 hollow balls used is 0.05-0.2wt% of the deionized water, the amount of sodium molybdate used is 200-300wt% of the CeO2 hollow balls used, the amount of thioacetamide used is 400-600wt% of the CeO2 hollow balls used, and the amount of polyethylene glycol monoester used is 40-90wt% of the sodium molybdate dihydrate; The efficacious agent solution includes a PTX solution and a PDL1i-containing solution. In the preparation of the carrier-efficacious agent, the carrier is first mixed with the PTX solution to obtain the carrier@PTX, and then the carrier@PTX is mixed with the PDL1i-containing solution to obtain the carrier@PTX@PDL1i, i.e., the carrier-efficacious agent.

2. The method for preparing biomimetic nanoparticles modified with cancer cell membranes according to claim 1, 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 based on PTX, and the amount of the carrier used is 100-300 wt % of the PTX.

3. The method for preparing biomimetic nanoparticles modified with cancer cell membranes according to claim 1, characterized in that: The cancer cell membrane suspension is prepared by adding cancer cell membranes into a PBS solution containing PMSF and resuspending the membranes. The pH of the PBS solution is 7-7.

8. The PBS solution contains 0.5-2 mM PMSF. The usage ratio of the cancer cell membranes to the PBS solution is 0.1-2 mg / mL.

4. The method for preparing biomimetic nanoparticles modified with cancer cell membranes according to claim 1, wherein: The usage amount of the cancer cell membrane is 80-120 wt % of the carrier-effective agent.

5. Biomimetic nanoparticles based on cancer cell membrane modification prepared by the preparation method according to any one of claims 1 to 4.

6. Use of the biomimetic nanoparticles modified with cancer cell membranes according to claim 5 in the preparation of anti-tumor drugs; the tumor is breast cancer.

7. An anti-tumor drug comprising the biomimetic nanoparticles modified based on cancer cell membranes according to claim 5; the tumor is breast cancer.

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