A biomimetic delivery system co-loading radionuclide and kdm5b degrader, and a preparation method and application thereof

Abstract

This invention relates to a biomimetic delivery system co-loaded with a radionuclide and a KDM5B degrading agent, its preparation method, and its application. The biomimetic delivery system includes liposomes, KDM5B degrading agent CMATACs encapsulated within the liposomes, and a radionuclide labeled on the surface of the liposomes. 177 Lu, erythrocyte membrane vesicles encapsulating the liposomes, and tumor homing peptide CREKA modified on the surface of the liposomes. Compared with the prior art, this invention achieves for the first time the synergistic delivery of "epigenetic regulation and targeted radionuclide therapy", and the prepared biomimetic delivery system has excellent in vivo anti-tumor effects and biosafety.

Description

Technical Field

[0001] This invention belongs to the field of nanomedicine delivery technology, and relates to a biomimetic delivery system co-loaded with a radionuclide and a KDM5B degrading agent, as well as its preparation method and application. Background Technology

[0002] Immune checkpoint blockade (ICB) therapy has made significant progress in the field of cancer treatment, but widespread drug resistance severely limits its clinical application. Studies have shown that the epigenetic regulator lysine-specific demethylase 5B (KDM5B) is negatively correlated with the expression levels of immune-activating genes and is a key target mediating immune escape.

[0003] Targeted therapy strategies for KDM5B mainly include small molecule inhibitors and protein degraders: (1) KDM5B small molecule inhibitor Several KDM5B small molecule inhibitors have been reported, but they face problems such as poor pharmacokinetic performance, insufficient specificity, and off-target toxicity. Traditional small molecule inhibitors require high doses to maintain effective concentrations, are prone to toxic side effects, and are difficult to overcome tumor drug resistance.

[0004] (2) Targeted protein degradation technology (PROTAC / CMATAC) Protein hydrolysis-targeting chimeras (PROTACs) and molecular chaperone-mediated autophagy-targeting chimeras (CMATACs) are novel protein degradation technologies developed in recent years. CMATACs possess advantages such as catalytic action mode, high selectivity, and high degradation efficiency, and can effectively downregulate KDM5B protein levels. However, CMATACs molecules themselves suffer from poor membrane permeability, insufficient in vivo stability, and low delivery efficiency at tumor sites, severely limiting their further application.

[0005] (3) Targeted radionuclide therapy (TRT) Targeted radionuclide therapy utilizes radiocouplers to specifically deliver radionuclides to tumor cells and their microenvironment, achieving precise irradiation of the tumor. 177Lu, due to its suitable beta-ray energy and half-life, has become a radionuclide with great application potential in the field of radiotherapy (TRT). However, the short residence time of 177Lu in tumor tissue and its severe toxic side effects remain major bottlenecks limiting its wider clinical application.

[0006] (4) Nanodelivery system Traditional liposomes are easily cleared by phagocytes of the reticuloendothelial system (RES), leading to large accumulations in the liver and spleen and reducing drug concentrations at tumor sites. Even after modification with polyethylene glycol (PEG), liposomes still cannot completely escape RES capture, and because they mainly rely on passive targeting, they may accumulate non-specifically in non-tumor tissues such as the liver and spleen, causing potential toxicity.

[0007] In summary, developing a biomimetic delivery system capable of efficiently co-delivering KDM5B degrading agents and radionuclides, with long circulation and active targeting capabilities, is of great significance for overcoming the limitations of existing therapies. Summary of the Invention

[0008] The purpose of this invention is to provide a biomimetic delivery system for co-loading radionuclides and KDM5B degrading agents, as well as its preparation method and application.

[0009] The objective of this invention can be achieved through the following technical solutions: One of the technical solutions of the present invention is to provide a biomimetic delivery system for co-loading a radionuclide and a KDM5B degrading agent, comprising liposomes, KDM5B degrading agent CMATACs encapsulated within the liposomes, and a radionuclide labeled on the surface of the liposomes. 177 Lu, red blood cell membrane vesicles encapsulating the liposomes, and tumor homing peptide CREKA modified on the surface of the liposomes.

[0010] In this invention, the biomimetic delivery system co-loaded with radionuclides and KDM5B degrading agents plays a synergistic role, manifested in two aspects: firstly, the CMATACs components effectively reverse the epigenetic immunosuppressive state of tumors and activate the cGAS-STING signaling pathway by selectively degrading KDM5B; secondly, 177 The β-rays released by Lu directly act on tumor cell DNA, inducing irreversible damage and exerting cytotoxic effects, further activating the cGAS-STING pathway. A significant synergistic effect is formed between the degradation of KDM5B and radiation-induced DNA breaks, both of which strongly activate the cGAS-STING signaling pathway, promote the secretion of type I interferon, and thus systematically enhance the anti-tumor immune response.

[0011] The second technical solution of the present invention is to provide a method for preparing a biomimetic delivery system as described in one of the above technical solutions, comprising the following steps: S1. Distearate phosphatidylethanolamine-polyethylene glycol-maleimide (DSPE-PEG-MAL) and tumor homing peptide CREKA are dissolved in an organic solvent and reacted at room temperature under inert gas protection. The reaction product DSPE-PEG-CREKA is then recovered by dialysis and drying. S2. Ursolic acid, dendritic polyethyleneimine, KDM5-C49, 1-ethyl-(3-dimethylaminopropyl)carbodiimine hydrochloride (EDC) and N-hydroxysuccinimide (NHS) were mixed in dimethyl sulfoxide and reacted with stirring at room temperature. After dialyzing and drying, KDM5B degradation agent CMATACs were obtained. S3, soybean lecithin, cholesterol, phospholipid polyethylene glycol dibenzocyclooctylene (DSPE-PEG-DBCO), DSPE-PEG-CREKA obtained in step S1, and CMATACs obtained in step S2 were dissolved in a mixed solution of dichloromethane and methanol. After removing the solvent, PBS buffer was added for hydration to obtain a nanoparticle solution. The solution was then repeatedly extruded through a liposome extruder to obtain LIPO-C nanoparticles. S4. Add the chelating agent DOTA-N3 to the mixture containing... 177 A sodium acetate solution of LuCl3 was reacted with stirring at high temperature to obtain... 177 Lu-DOTA-N3; The LIPO-C nanoparticles obtained in step S3, 177 The Lu-DOTA-N3 mixture reacts to obtain 177 Lu-labeled liposomes 177 Lu-LIPO-C; S5. Combine the red blood cell membrane vesicle suspension with the solution obtained in step S4. 177 Lu-LIPO-C mixtures are repeatedly extruded through a liposome extruder to obtain hybrid nanovesicles, which are the biomimetic delivery system CELLC.

[0012] In some specific embodiments, in step S1, the mass ratio of distearate phosphatidylethanolamine-polyethylene glycol-maleimide to tumor homing peptide CREKA is (7~10):(1~3). The molecular weight cutoff for dialysis is 1000 Da.

[0013] As a more preferred embodiment, the mass ratio of distearate phosphatidylethanolamine-polyethylene glycol-maleimide to tumor homing peptide CREKA is 8.5:1.78.

[0014] In this invention, the modification amount of the tumor homing peptide CREKA is 1-10% of the total phospholipid content for DSPE-PEG-CREKA.

[0015] In some specific embodiments, in step S2, the mass ratio of ursolic acid, dendritic polyethyleneimine, KDM5-C49, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide is (0.5~3):(0.5~1.5):(0.5~1.5):(15~20):(1~3). The molecular weight cutoff for dialysis is 2000 Da.

[0016] As a more preferred embodiment, the mass ratio of ursolic acid, dendritic polyethyleneimine, KDM5-C49, 1-ethyl-(3-dimethylaminopropyl)carbodiimine hydrochloride and N-hydroxysuccinimide is 1.37:0.9:0.62:19.2:2.3.

[0017] In some specific embodiments, in step S3, the ratio of soybean lecithin, cholesterol, phospholipid polyethylene glycol dibenzocyclooctylene, DSPE-PEG-CREKA, and CMATACs is (10~15) mg: (2~4) mg: (0.5~1.5) mg: (0.5~1.5) mg: (0.1 μM~3 μM). The volume ratio of dichloromethane to methanol is 2:1.

[0018] As a more preferred option, the ratio of soybean lecithin, cholesterol, phospholipid polyethylene glycol dibenzocyclooctylene, DSPE-PEG-CREKA, and CMATACs is 12 mg: 3 mg: 1 mg: 1 mg: (0.1 μM~2 μM).

[0019] In some specific embodiments, in step S3, the temperature of the hydration reaction is (45~55)℃ and the time is (15~30)min; The filter membrane of the liposome extruder has a pore size of 200 nm.

[0020] In some specific embodiments, step S4 involves the chelating agent DOTA-N3, containing... 177 The ratio of LuCl3 to sodium acetate solution is (0.1~0.5) mg : (90~120) μL, in which radioactive nuclides... 177 The radioactive dose of Lu is 50-300 μCi; The temperature of the high-temperature stirring reaction is (85~100)℃ and the time is (10~20)min.

[0021] As a more preferred option, the chelating agent DOTA-N3 contains... 177 The ratio of LuCl3 to sodium acetate solution was 0.25 mg: 100 μL, in which radioactive nuclides were present. 177 The radioactive dose of Lu is 50-300 μCi.

[0022] In some specific embodiments, step S4 involves LIPO-C nanoparticles, 177 The ratio of Lu-DOTA-N3 is (0.1~0.5) mg : (0.5~3) mL; The temperature of the mixed reaction was (35~39)℃ and the time was (1.5~2.5) h.

[0023] As a better alternative, LIPO-C nanoparticles, 177 The ratio of Lu-DOTA-N3 was 0.25 mg: 1 mL.

[0024] In some specific embodiments, in step S5, the red blood cell membrane vesicle suspension and 177 Lu-LIPO-C is mixed at a protein-phospholipid mass ratio of 1:2 to 2:1; The filter membrane of the liposome extruder has a pore size of 400 nm and 200 nm.

[0025] As a more preferred option, the red blood cell membrane vesicle suspension and 177 Lu-LIPO-C is mixed at a protein-phospholipid mass ratio of 1:1.

[0026] The third technical solution of the present invention is to provide an application of the biomimetic delivery system as described in the second technical solution above in the preparation of antitumor drugs.

[0027] In some specific embodiments, the tumor is breast cancer.

[0028] Compared with the prior art, the present invention has the following advantages: (1) For the first time, the synergistic delivery of "epigenetic regulation and targeted radionuclide therapy" was achieved. This invention is the first to combine CMATACs-based epigenetic regulatory strategies with 177 Lu-labeled targeted radionuclide therapy was organically combined. Cell experiments confirmed that CMATACs could degrade KDM5B protein in a concentration-dependent manner. At a treatment concentration of 2 μM, the expression level of KDM5B in the liposome-encapsulated CMATACs group was reduced by approximately 35% compared with that in the free drug group (P<0.05). 177 The Lu-labeled liposomes achieved a radiochemical purity of over 98%, and maintained a labeling rate of over 76.9% after incubation in physiological saline and serum systems for 108 h.

[0029] (2) Excellent in vivo antitumor effect and biosafety In the 4T1 subcutaneous tumor-bearing mouse model, the biomimetic delivery system of this invention achieved a tumor growth inhibition rate of 82.8%, significantly superior to other treatment groups. No significant decrease in body weight was observed in any group of mice during treatment, indicating that the nanocomposite has good in vivo biocompatibility.

[0030] (3) Reduce usage costs and facilitate clinical translation The raw materials used in this invention are all domestically produced, which can significantly reduce the cost of mass cytometry and related technologies. Compared with traditional liposomes, the biomimetic modification of the erythrocyte membrane significantly prolongs the in vivo circulation time of the drug, reduces the frequency of administration, and lowers toxic side effects. Attached Figure Description

[0031] Figure 1 The synthetic route for CMATACs is shown.

[0032] Figure 2 for 177 A schematic diagram of the structure of Lu-DOTA-N3.

[0033] Figure 3 A represents TEM images of liposomes, erythrocyte membrane vesicles, and erythrocyte membrane-liposome hybrid nanovesicles, respectively. Figure 3 B represents the particle size statistics for liposomes, erythrocyte membrane vesicles, and erythrocyte membrane-liposome hybrid nanovesicles, respectively.

[0034] Figure 4 This study validated the fluorescence colocalization and FRET of the CELLC biomimetic liposome nanocomposite for erythrocyte membranes.

[0035] Figure 5 A represents the immunoblotting results of KDM5B protein after treatment of 4T1 cells in the CMATAC group and the LIPO-CMATAC group. Figure 5 B represents the statistical data from graph A.

[0036] Figure 6 A is a radioactive nuclide 177 Lu labeling rate; Figure 6 B is a radioactive nuclide. 177 Lu stability results.

[0037] Figure 7 A is a schematic diagram of the mouse model and drug administration process; Figure 7 B represents the tumor volume in the mouse model after treatment with different drug groups. Detailed Implementation

[0038] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. These embodiments are based on the technical solution of the present invention and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0040] Unless otherwise specified, the materials and processes used in the following embodiments or examples are conventional materials and processes employed in the art to achieve the corresponding functions.

[0041] Example 1: This embodiment provides a biomimetic delivery system CELLC that co-loads a radionuclide and a KDM5B degrading agent, namely a erythrocyte membrane biomimetic liposome nanocomposite, the structure of which includes: (1) Liposome core: Liposomes composed of soybean lecithin and cholesterol, internally encapsulated with KDM5B degrading agent CMATACs; (2) Radionuclide labeling: Radionuclides are chelated by the macrocyclic ligand DOTA-N3. 177 Lu, labeled on the surface of liposomes; (3) Red blood cell membrane shell: Natural red blood cell membrane vesicles coated on the outside of liposomes by co-extrusion fusion method, giving the system immune escape and long circulation characteristics; (4) Targeted modification: The liposome surface is modified with tumor homing peptide CREKA (Cys-Arg-Glu-Lys-Ala) to actively target the fibroin-fibronectin complex that is highly expressed in the tumor microenvironment.

[0042] Example 2: This embodiment provides a method for preparing the CELLC biomimetic delivery system of Embodiment 1 above, including the following steps: (1) Synthesis of DSPE-PEG-CREKA 8.5 mg of distearate-phosphatidylethanolamine-polyethylene glycol-maleimide (DSPE-PEG-MAL) and 1.78 mg of CREKA were dissolved together in a 10% methanol solution, and the mixture was stirred at room temperature for 4 h under nitrogen protection. After the reaction was completed, the mixture was transferred to a dialysis bag with a molecular weight cutoff of 1000 Da, and dialyzed with distilled water for 24 h, changing the distilled water every 1 h. After dialysis, the liquid in the dialysis bag was collected, and the reaction product was recovered by freeze-drying to obtain DSPE-PEG-CREKA.

[0043] (2) Synthesis of CMATACs like Figure 1The synthetic route shown involved weighing appropriate amounts of ursolic acid, dendritic polyethyleneimine, KDM5-C49, 1-ethyl-(3-dimethylaminopropyl)carbodiimine hydrochloride (EDC), and N-hydroxysuccinimide (NHS), each dissolved in dimethyl sulfoxide. The final concentrations of ursolic acid, dendritic polyethyleneimine, KDM5-C49, and NHS were 10 mg / mL, and the concentration of EDC was 100 mg / mL. 137 µL of ursolic acid solution, 90 µL of dendritic polyethyleneimine solution, 62 µL of KDM5-C49 solution, 192 µL of EDC solution, 230 µL of NHS solution, and 152 µL of dimethyl sulfoxide were sequentially added to a reaction flask. The reaction was carried out under light-protected conditions with magnetic stirring at room temperature for 20 h. After the reaction was complete, the mixture was dialyzed against dimethyl sulfoxide for 1 h using a 2000 kDa dialysis bag, followed by dialyzed against ultrapure water overnight. The dialysis product was collected, freeze-dried, and weighed to obtain CMATACs.

[0044] (3) Preparation of liposomes 12 mg of soybean lecithin, 3 mg of cholesterol, 1 mg of phospholipid polyethylene glycol dibenzocyclooctylene (DSPE-PEG-DBCO), 1 mg of DSPE-PEG-CREKA, and different concentrations (0, 0.5, 1, 2 μM) of CMATAC were dissolved in a mixture of 10 mL of dichloromethane and 5 mL of methanol. The solution was transferred to a round-bottom flask, and the solvent was removed by rotary evaporation, forming a white transparent film at the bottom of the flask. 4 mL of PBS buffer was added to the round-bottom flask, and the solution was hydrated at 50 °C for 20 min. The resulting nanoparticle solution was repeatedly extruded 20 times using a liposome extruder with a 200 nm pore size to obtain LIPO-C nanoparticles.

[0045] (4) 177 Lu's mark Weigh 0.25 mg of DOTA-N3 and add it to a container containing... 177 100 μL sodium acetate solution of LuCl3 (pH 5.6, radioactive nuclide) 177 The radioactive dose of Lu was 200 μCi / , and the mixture was placed in a constant temperature mixer and incubated at 95°C and 800 rpm for 15 min to obtain... 177 Lu-DOTA-N3, for example Figure 2 The structure shown.

[0046] Weigh 1 mL of liposome solution (containing DSPE-PEG-DBCO) dispersed in PBS buffer, add 0.25 mg of... 177Lu-labeled DOTA-N3 (177Lu-DOTA-N3) was placed in a thermostatic mixer and incubated with shaking at 37°C and 800 rpm for 2 h to obtain... 177 Lu-labeled liposomes ( 177 Lu-LIPO-C).

[0047] The prepared 177 Lu-LIPO-C was incubated in physiological saline and serum systems for 108 h, respectively. The radiolabeling rate was determined by radioactive thin-layer chromatography, and the results are as follows: Figure 6 As shown, 177 The Lu-labeled liposomes achieved a radiochemical purity of over 98%, and maintained a labeling rate of over 76.9% after incubation in physiological saline and serum systems for 108 h.

[0048] (5) Extraction of red blood cell membranes Blood was collected from the orbital vein or heart of mice using a syringe and transferred to a centrifuge tube containing an appropriate amount of heparin sodium anticoagulant. The blood was gently inverted to mix. The anticoagulated blood was transferred to a new centrifuge tube and centrifuged at 1500 rpm for 10 min at 4°C. After centrifugation, the supernatant was carefully aspirated, retaining the lower red blood cell layer. Five volumes of pre-chilled physiological saline were added to the centrifuge tube containing red blood cells, and the mixture was gently pipetted to mix. The tube was centrifuged at 3000 rpm for 10 min at 4°C, and the supernatant was discarded. This washing step was repeated three times. The washed red blood cells were transferred to another centrifuge tube, and pre-chilled hypotonic 0.25×PBS buffer was added at a ratio of 1:30 (red blood cell volume). The mixture was gently stirred and incubated on ice for 30 min. After hemolysis, the tube was centrifuged at 10000 rpm for 20 min at 4°C, and the supernatant was discarded. The red blood cell membrane precipitate was collected. Add an appropriate amount of pre-cooled PBS to the precipitate, gently resuspend by pipetting, centrifuge at 10000-12000 r / min for 20-30 min at 4℃, discard the supernatant, and repeat the washing process 3 times. Resuspend the red blood cell membrane precipitate after the last wash in an appropriate amount of PBS and store it at -80℃ for later use.

[0049] (6) Membrane fusion fabrication of biomimetic delivery system CELLC Erythrocyte membrane-liposome hybrid nanovesicles were prepared using a co-extrusion fusion method. The prepared erythrocyte membrane vesicle suspension and drug-loaded liposome suspension were mixed at a protein-phospholipid ratio of 1:1 (w / w) and thoroughly mixed using a vortex mixer. The mixture was transferred to a liposome extruder and repeatedly extruded through 400 nm and 200 nm polycarbonate membranes. Co-extrusion cycles were performed at room temperature for at least 10 cycles to allow the two types of vesicles to fuse fully under mechanical force, forming hybrid nanovesicles with uniform particle size. The resulting product, the erythrocyte membrane-liposome hybrid nanovesicle suspension, is the biomimetic delivery system CELLC, which can be stored at 4°C for short-term storage or further characterization.

[0050] The prepared biomimetic delivery system CELLC was characterized and its performance was tested as follows: (1) such as Figure 3 The images shown are TEM images and particle size statistics of liposomes, erythrocyte membrane vesicles, and erythrocyte membrane-liposome hybrid nanovesicles (CELLC).

[0051] (2) For example Figure 4 As shown, this illustrates the fluorescence colocalization of erythrocyte membrane-liposome hybrid nanovesicles (CELLC).

[0052] (3) Verification of KDM5B degradation effect at the cellular level Mouse breast cancer 4T1 cells were injected at a rate of 2 × 10⁻⁶ 5 Cells were seeded at a density of cells / well in six-well plates. After the cells adhered, 0, 0.5, 1, and 2 μM of free drug (CMATAC group) and liposomes containing CMATAC (LIPO-CMATAC group) were added and incubated with the cells for 24 h.

[0053] Cells were lysed on ice using RIPA cell lysis buffer containing protease and phosphatase inhibitors, and protein solutions were collected. Protein samples were separated using 8% polyacrylamide gel electrophoresis, transferred to PVDF membranes, and detected by Western blotting using Anti-KDM5B antibody.

[0054] The results are as follows Figure 5 The results showed that both drugs could reduce the expression level of KDM5B in 4T1 cells in a concentration-dependent manner. At a treatment concentration of 2 μM, the expression level of KDM5B in the LIPO-CMATAC group was reduced by about 35% compared with that in the free drug group.

[0055] (4) In vivo anti-tumor experiment Pre-select 4T1 cells in logarithmic growth phase according to 10 6 Cells were seeded at a density of 1 cell per mouse into the legs of 8-week-old female BALB / c mice. When the 4T1 subcutaneous tumor volume reached 100 mm... 3 When that time is reached, record that time point as day 0.

[0056] The mice were randomly divided into 6 groups (n=5 in each group): G1: Saline group; G2: 177 Lu-LIPO group (radioactive nuclides) 177 Lu-labeled liposomes); G3: 177 Lu-LIPO-C group (radioactive nuclides) 177 Lu-labeled and CREKA-modified liposomes); G4: 177 Lu-LIPO-CR group (radioactive nuclides) 177 G5: LIPO-CCR group (liposomes-erythrocyte membrane fusion loaded with CMATACs and modified with CREKA); G6; 177 Lu-LIPO-CCR group (i.e., CELLC group, 0.5 mg / kg, containing radionuclides) 177 The radioactive dose of Lu is 300 μCi / animal, and the CMATACs loading is 2 μM.

[0057] On days 0 and 2, mice in each group were injected with the corresponding drugs via the tail vein. Starting from day 0, the body weight and tumor length and short diameter of the mice were measured every other day, and the results were recorded according to the formula V=ab. 2 / 2 Calculate tumor volume. When the tumor volume of any group of mice reaches 2000 mm... 3 All mice were euthanized and analyzed.

[0058] The tumor volume was measured, and the results were as follows: Figure 7 As shown, the tumor growth inhibition rate in the G6 group reached 82.8%, which was significantly higher than that in other drug treatment groups.

[0059] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. A biomimetic delivery system for co-loading radionuclides and KDM5B degrading agents, characterized in that, Includes liposomes, KDM5B degrading agents CMATACs encapsulated within the liposomes, and radionuclides labeled on the surface of the liposomes. 177 Lu, red blood cell membrane vesicles encapsulating the liposomes, and tumor homing peptide CREKA modified on the surface of the liposomes.

2. A biomimetic delivery system for co-loading radionuclides and KDM5B degrading agent as described in claim 1, characterized in that, Includes the following steps: S1. Distearate phosphatidylethanolamine-polyethylene glycol-maleimide and tumor homing peptide CREKA are dissolved in an organic solvent and reacted at room temperature under inert gas protection. The reaction product DSPE-PEG-CREKA is then recovered by dialysis and drying. S2. Ursolic acid, dendritic polyethyleneimine, KDM5-C49, 1-ethyl-(3-dimethylaminopropyl)carbodiimine hydrochloride and N-hydroxysuccinimide were mixed in dimethyl sulfoxide, stirred at room temperature, dialyzed and dried to obtain KDM5B degradation agent CMATACs. S3, soybean lecithin, cholesterol, phospholipid polyethylene glycol dibenzocyclooctylene, DSPE-PEG-CREKA obtained in step S1 and CMATACs obtained in step S2 were dissolved in a mixed solution of dichloromethane and methanol. After removing the solvent, PBS buffer was added for hydration reaction to obtain nanoparticle solution. The solution was then repeatedly extruded through a liposome extruder to obtain LIPO-C nanoparticles. S4. Add the chelating agent DOTA-N3 to the mixture containing... 177 A sodium acetate solution of LuCl3 was reacted with stirring at high temperature to obtain... 177 Lu-DOTA-N3; The LIPO-C nanoparticles obtained in step S3, 177 The Lu-DOTA-N3 mixture reacts to obtain 177 Lu-labeled liposomes 177 Lu-LIPO-C; S5. Combine the red blood cell membrane vesicle suspension with the solution obtained in step S4. 177 Lu-LIPO-C mixtures are repeatedly extruded through a liposome extruder to obtain hybrid nanovesicles, which are the biomimetic delivery system CELLC.

3. The preparation method of the biomimetic delivery system for co-loaded radionuclides and KDM5B degrading agent as described in claim 2, characterized in that, In step S1, the mass ratio of distearate phosphatidylethanolamine-polyethylene glycol-maleimide to tumor homing peptide CREKA is (7~10):(1~3). The molecular weight cutoff for dialysis is 1000 Da.

4. The preparation method of the biomimetic delivery system for co-loaded radionuclides and KDM5B degrading agent as described in claim 2, characterized in that, In step S2, the mass ratio of ursolic acid, dendritic polyethyleneimine, KDM5-C49, 1-ethyl-(3-dimethylaminopropyl)carbodiimine hydrochloride, and N-hydroxysuccinimide is (0.5~3):(0.5~1.5):(0.5~1.5):(15~20):(1~3). The molecular weight cutoff for dialysis is 2000 Da.

5. The preparation method of the biomimetic delivery system for co-loaded radionuclides and KDM5B degrading agent as described in claim 2, characterized in that, In step S3, the ratio of soybean lecithin, cholesterol, phospholipid polyethylene glycol dibenzocyclooctylene, DSPE-PEG-CREKA, and CMATACs is (10~15) mg: (2~4) mg: (0.5~1.5) mg: (0.5~1.5) mg: (0.1 μM~2 μM). The volume ratio of dichloromethane to methanol is 2:1; The hydration reaction was carried out at a temperature of (45~55)℃ and for a time of (15~30) min. The filter membrane of the liposome extruder has a pore size of 200 nm.

6. The method for preparing the biomimetic delivery system for co-loaded radionuclides and KDM5B degrading agent as described in claim 2, characterized in that, In step S3, the hydration reaction is carried out at a temperature of (45~55)℃ and a time of (15~30) min. The filter membrane of the liposome extruder has a pore size of 200 nm.

7. The preparation method of the biomimetic delivery system for co-loaded radionuclides and KDM5B degrading agent as described in claim 2, characterized in that, Step S4, chelating agent DOTA-N3, containing... 177 The ratio of LuCl3 to sodium acetate solution is (0.1~0.5) mg : (90~120) μL, in which radioactive nuclides... 177 The radioactive dose of Lu is 50-300 μCi; The temperature of the high-temperature stirring reaction is (85~100)℃ and the time is (10~20)min.

8. The method for preparing the biomimetic delivery system for co-loaded radionuclides and KDM5B degrading agent as described in claim 2, characterized in that, Step S4, LIPO-C nanoparticles, 177 The ratio of Lu-DOTA-N3 is (0.1~0.5) mg : (0.5~3) mL; The temperature of the mixed reaction was (35~39)℃ and the time was (1.5~2.5) h.

9. The method for preparing the biomimetic delivery system for co-loaded radionuclides and KDM5B degrading agent as described in claim 2, characterized in that, Step S5, red blood cell membrane vesicle suspension and 177 Lu-LIPO-C is mixed at a protein-phospholipid mass ratio of 1:2 to 2:1; The filter membrane of the liposome extruder has a pore size of 400 nm and 200 nm.

10. The application of the biomimetic delivery system for co-loaded radionuclides and KDM5B degrading agents as described in claim 1 in the preparation of antitumor drugs.

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