Preparation method and application of cell membrane bionic polymer nano-drug

Abstract

The invention relates to the technical field of drug research and development, and discloses a cell membrane bionic polymer nano-drug and a preparation method and application thereof.The nano-drug comprises a cell membrane and polymer nanoparticles, and the polymer nanoparticles are composed of polymers, positively charged metal peroxide nanoparticles and chemotherapeutic drugs; the structural formula of the polymer is as follows: # imgabs0 #, wherein the polyethyleneimine in the polymer is polyethyleneimine with the weight-average molecular weight of 1000 to 10000, and n is equal to 3 to 30; the polyethylene glycol in the polymer is polyethylene glycol with the weight-average molecular weight of 2000-10000, and m is equal to 50-250. The nano-drug obtained by the invention can be enriched at a tumor site, and the tumor cell membrane is damaged by generating hydrogen peroxide in situ on the surface of the tumor cell membrane, so that the biological barrier of the tumor cell membrane, which hinders drug transport, is broken through, and the anti-tumor drug effect of the chemotherapeutic drug is enhanced.

Description

Technical Field

[0001] The present invention relates to the technical field of drug research and development, and in particular to a preparation method of a cell membrane bionic polymer nano drug and its application. Background Art

[0002] Cancer is the second leading cause of death worldwide, bringing a heavy economic burden to individuals and society. To this day, chemotherapy is still one of the main ways to treat cancer. The emergence of nanotechnology has brought new breakthroughs in chemotherapy: polymer micelles, liposomes, cell membrane vesicles, and inorganic nanoparticles, etc., can effectively improve the safety and bioavailability of drugs through their unique physicochemical properties. However, the in vivo utilization of nanomedicines is still significantly insufficient. Only less than 1% of nanomedicines can accumulate at the tumor site, and less than 0.01% of nanomedicines can interact with tumor cells and enter cells. Nanomedicines are generally endocytosed by cells when they come into contact with cell membranes, and then transported to lysosomes for degradation. Only some drugs that escape lysosomes can exert their efficacy. Even if nanomedicines successfully escape lysosomes and release chemotherapeutic drugs, they may still be destroyed by tumor cells through the autophagy-lysosome pathway or excreted by drug resistance-related proteins. In addition, the endocytosis efficiency of nanomedicines by tumor cells is also affected by complex factors such as nanocarrier morphology, size, material, surface charge or modification, and cell type, which makes the conditions for efficient endocytosis extremely harsh. These characteristics make the tumor cell membrane the main obstacle for nanomedicines to enter tumor cells, and an important factor affecting the low effective utilization rate of nanomedicines. Therefore, it is necessary to design and construct appropriate nanomedicines to break through the biological barrier of tumor cell membranes, improve drug utilization, and enhance chemotherapy effects. Summary of the invention

[0003] In order to solve the above technical problems, the present invention provides a method for preparing a cell membrane biomimetic polymer nanomedicine and its application. The nanomedicine obtained by the present invention can be enriched in the tumor site, and the tumor cell membrane is damaged by in situ generation of hydrogen peroxide (H2O2) on the surface of the tumor cell membrane, thereby breaking through the tumor cell membrane, which is a biological barrier that hinders drug transport, and promoting the loaded chemotherapy drugs to directly enter the tumor cells through the holes in the cell membrane, thereby increasing the concentration of effective chemotherapy drugs in the tumor cells, thereby enhancing the anti-tumor efficacy of the chemotherapy drugs.

[0004] The present invention provides a cell membrane biomimetic polymer nanomedicine, the nanomedicine comprises a cell membrane and polymer nanoparticles, the polymer nanoparticles are composed of a polymer, positively charged metal peroxide nanoparticles and a chemotherapeutic drug; The structural formula of the polymer is as follows: ; The polyethyleneimine in the polymer is polyethyleneimine with a weight average molecular weight of 1000-10000, and n=3-30; the polyethylene glycol in the polymer is polyethylene glycol with a weight average molecular weight of 2000-10000, and m=50-250.

[0005] Furthermore, the metal peroxide includes one or more of calcium peroxide, copper peroxide, iron oxide and manganese dioxide.

[0006] Furthermore, the chemotherapy drugs include one or more of doxorubicin, camptothecin, paclitaxel and gemcitabine.

[0007] Furthermore, the preparation method of the positively charged metal peroxide nanoparticles comprises: The metal dichloride, ammonia water and pentalysine are dissolved in an ethanol solution, and a hydrogen peroxide solution is added, reacted, and centrifuged to obtain a solid precipitate; the solid matter is then placed in a saturated carbon dioxide environment to obtain positively charged metal peroxide nanoparticles.

[0008] Furthermore, the metal dichloride includes one or more of calcium chloride, cupric chloride, ferrous chloride and manganese chloride.

[0009] Furthermore, the ethanol solution is prepared by mixing anhydrous ethanol with deionized water, wherein the volume ratio of anhydrous ethanol to deionized water is (5-29):1.

[0010] Furthermore, the mass concentration of the metal dichloride in the ethanol solution is 1%-30%.

[0011] Furthermore, the mass concentration of the hydrogen peroxide solution is 3%.

[0012] Furthermore, the ratio of the metal dichloride to pentalysine is (10-100):1 by mass.

[0013] Furthermore, the ratio of the aqueous ammonia, the ethanol solution and the hydrogen peroxide solution is 1:(100-1000):2 by volume.

[0014] Furthermore, the reaction time is 1 min-20 min.

[0015] Furthermore, those skilled in the art should understand that in order to promote sufficient reaction, stirring may be performed during the reaction.

[0016] Furthermore, the standing time is 1.5h-4h.

[0017] Furthermore, the cell membrane is a tumor cell.

[0018] The present invention also provides a method for preparing the cell membrane biomimetic polymer nanomedicine, comprising the following steps: The polymer and positively charged metal peroxide nanoparticles are dissolved and dispersed in deionized water, and an organic solvent containing a chemotherapeutic drug is added, dialyzed, and freeze-dried to obtain polymer nanoparticles; the cell membrane and the polymer nanoparticles are mixed, and centrifuged and ultrafiltered through a micro-extruder with a polycarbonate membrane to obtain a cell membrane biomimetic polymer nanomedicine.

[0019] Furthermore, the organic solvent includes one or more of dimethyl sulfoxide, methanol and dimethylformamide.

[0020] Furthermore, the mass concentration of the chemotherapy drug in the organic solvent is 0.5 mg / mL-5 mg / mL.

[0021] Furthermore, the specific operation of the dialysis includes: placing the target object in a dialysis bag, and then placing the dialysis bag in deionized water for dialysis.

[0022] Furthermore, the dialysis time is 2h-6h.

[0023] Furthermore, the ratio of the polymer to the positively charged metal peroxide nanoparticles is (3-30):1 by mass.

[0024] Furthermore, the mass concentration of the polymer in deionized water after being dissolved and dispersed is 3 mg / mL-30 mg / mL.

[0025] Furthermore, the ratio of the dissolved and dispersed deionized water to the organic solvent containing the chemotherapeutic drug is (10-100):1 by volume.

[0026] Furthermore, the mass ratio of the cell membrane to the polymer nanoparticles is 1:(10-50).

[0027] Furthermore, the specific process of passing through the micro-extruder of the polycarbonate membrane is to sequentially pass through the micro-extruder with polycarbonate membranes with a pore size of 400 nm and a pore size of 200 nm.

[0028] Furthermore, the preparation method of the polymer comprises: Step S1: dissolving 16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)hexadecanoic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mixing well, adding polyethylene glycol-polyethyleneimine and N,N-diisopropylethylamine, reacting, dialyzing, and freeze-drying to obtain compound a; Step S2: dissolving 4-carboxyphenylboronic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mixing evenly, adding compound a and N,N-diisopropylethylamine, reacting, adding diethylamine, dialyzing, and freeze-drying to obtain compound b; Step S3: dissolve 3-(diethylamino)propionic acid hydrochloride, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mix well, add compound b and N,N-diisopropylethylamine, dialyze, and freeze-dry to obtain a polymer.

[0029] Furthermore, in the step S1, the polyethylene glycol in the polyethylene glycol-polyethylene imine is polyethylene glycol with a weight average molecular weight of 2000-10000, and the polyethylene imine is polyethylene imine with a weight average molecular weight of 1000-10000.

[0030] In the present invention, those skilled in the art can select any polyethylene glycol with a weight average molecular weight in the range of 1000-10000 and any polyethyleneimine with a weight average molecular weight in the range of 2000-10000 to combine, and synthesize the polyethylene glycol-polyethyleneimine material by conventional means in the art.

[0031] Furthermore, in step S1, the ratio of 16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)hexadecanoic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is (1-2):(1-2):1 by mass.

[0032] Furthermore, in the step S1, before uniform mixing, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(1000-10000) by volume.

[0033] Furthermore, in step S1, the mass concentration of 1-hydroxybenzotriazole in dimethylformamide is 0.5 mg / mL-5 mg / mL.

[0034] Furthermore, in the step S1, there are many ways to mix evenly. In the present invention, stirring is adopted. As an example, the stirring time is 20 minutes.

[0035] Furthermore, in the step S1, the mass ratio of the polyethylene glycol-polyethyleneimine to 1-hydroxybenzotriazole is (10-100):1.

[0036] Furthermore, in step S1, the volume ratio of N,N-diisopropylethylamine added before and after mixing is (1-2):(1-5).

[0037] Furthermore, in step S1, the reaction time is 1 h-3 h.

[0038] Furthermore, in step S1, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days.

[0039] Furthermore, in the mixed solution of dimethylformamide and deionized water, the volume ratio of dimethylformamide to deionized water is 1:(100-1000).

[0040] Furthermore, in step S2, the ratio of 4-carboxyphenylboronic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is 1:(2-4):(2-4) by mass.

[0041] Furthermore, in the step S2, before uniform mixing, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(50-200) by volume.

[0042] Furthermore, in step S2, the mass concentration of 4-carboxyphenylboronic acid in dimethylformamide is 3 mg / mL-10 mg / mL.

[0043] Furthermore, in step S2, there are many ways to achieve uniform mixing. In the present invention, stirring is used. As an example, the stirring time is 20 minutes.

[0044] Furthermore, in step S2, the volume ratio of N,N-diisopropylethylamine added before and after mixing is 1:1.

[0045] Furthermore, in step S2, the mass concentration of compound a in dimethylformamide is 0.1 mg / mL-1 mg / mL.

[0046] Furthermore, in step S2, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days.

[0047] Furthermore, in the mixed solution of dimethylformamide and deionized water, the volume ratio of dimethylformamide to deionized water is 1:(100-1000).

[0048] Furthermore, in step S3, the ratio of 3-(diethylamino)propionic acid hydrochloride, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is 1:(2-10):(2-10) by mass.

[0049] Furthermore, in step S3, before uniform mixing, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(1000-10000) by volume.

[0050] Furthermore, in step S3, the mass concentration of 3-(diethylamino)propionic acid hydrochloride in dimethylformamide is 0.05 mg / mL-1 mg / mL.

[0051] Furthermore, in step S3, there are many ways to achieve uniform mixing. In the present invention, stirring is used. As an example, the stirring time is 20 minutes.

[0052] Furthermore, in step S3, the volume ratio of N,N-diisopropylethylamine added before and after mixing is (1-2):(1-2).

[0053] Furthermore, in step S3, the mass concentration of compound b in dimethylformamide is 1 mg / mL-10 mg / mL.

[0054] Furthermore, in step S3, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days.

[0055] Furthermore, in the mixed solution of dimethylformamide and deionized water, the volume ratio of dimethylformamide to deionized water is 1:(100-1000).

[0056] The invention also provides application of the cell membrane bionic polymer nanomedicine in treating cancer.

[0057] The embodiments of the present invention have the following technical effects: In the cell membrane biomimetic polymer nanomedicine of the present invention, firstly, the cell membrane can recognize homologous tumor cells, which is conducive to the enrichment of the cell membrane biomimetic polymer nanomedicine in the tumor site; in view of the acidic microenvironment (pH 6.5) in the tumor tissue, the polymer in the cell membrane biomimetic polymer nanomedicine will swell and release the loaded metal peroxide nanoparticles, which then adhere to the surface of the tumor cells, generate H2O2 in situ and cause tumor cell membrane perforation. Subsequently, intracellular adenosine triphosphate (ATP) begins to seep out through these transmembrane pores, further leading to the dissociation of the cell membrane biomimetic polymer nanomedicine, thereby releasing chemotherapy drugs, which directly enter the tumor cells through the damaged cell membrane and induce cell apoptosis. The present invention provides a strategy for efficiently delivering drugs to tumor cells to eliminate tumor cells by breaking through the cell membrane biological barrier.

[0058] 2. In order to ensure that the metal peroxide can stably play a role in the tumor cell membrane, polylysine is adsorbed on the surface of the metal peroxide in the present invention, so that the metal peroxide is positively charged. Since the positively charged metal peroxides have the same charge, they can be evenly dispersed in the polymer nanoparticles to reduce agglomeration; on the other hand, they are conducive to adsorption on the surface of the tumor cell membrane.

[0059] 3. The cell membrane biomimetic polymer nanomedicine of the present invention has a simple preparation process, mild conditions, good biocompatibility and stability, and can effectively release drugs on demand in response to pH and ATP changes in the tumor microenvironment.

[0060] 4. The cell membrane biomimetic polymer nanomedicine provided by the present invention can effectively load water-insoluble pharmacologically active molecules such as doxorubicin. The particle size of the nanomedicine is uniform, ranging from 10nm to 300nm, which greatly improves the solubility of poorly soluble drugs. The nanomedicine can be enriched in tumor lesions through the enhancement and retention (EPR) effect and the homologous targeting ability of the tumor cell membrane, and damage the tumor cell membrane and release drugs in response to changes in the tumor microenvironment. The physically loaded drugs are quickly released from the nanoparticles and directly enter the tumor cells through the transmembrane pores, thereby increasing the effective drug concentration in the tumor, thereby exerting a highly efficient tumor cell killing effect. The cell membrane biomimetic polymer nanomedicine provided by the present invention can effectively avoid capture by the reticuloendothelial system in the body, and has both passive and active targeting, which is beneficial to improving the utilization rate of the drug under intravenous administration. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0062] Figure 1 It is the hydrated particle size diagram and transmission electron microscope TEM image of the intermediate product of Example 1, which is the positively charged calcium peroxide nanoparticles.

[0063] Figure 2 It is a hydrated particle size diagram and TEM diagram of the intermediate product polymer nanoparticles of Example 1 of the present invention.

[0064] Figure 3 It is a hydrated particle size diagram and TEM diagram of the cell membrane biomimetic polymer nanomedicine constructed in Example 1 of the present invention.

[0065] Figure 4 It is a diagram of the Zeta potential change of the positively charged calcium peroxide nanoparticles of the intermediate product of Example 1 of the present invention, Comparative Example 1, the intermediate product polymer nanoparticles in Example 1 and Example 1.

[0066] Figure 5 It is the ultraviolet absorption spectrum of the cell membrane biomimetic polymer nanomedicine constructed by the present invention.

[0067] Figure 6 The method is to detect the complete retention of the tumor cell membrane in the cell membrane biomimetic polymer nanomedicine constructed by the present invention by protein blotting.

[0068] Figure 7 It is the in vitro drug release curve of the cell membrane biomimetic polymer nanomedicine constructed by the present invention.

[0069] Figure 8 This is a laser confocal microscope image of the co-culture of the cell membrane biomimetic polymer nanomedicine constructed by the present invention and mouse 4T1 tumor cells, with a scale of 5 μm. Figure 8 a in the middle is the cell nucleus, Figure 8 Figure b shows the distribution of positively charged calcium peroxide nanoparticles in cell membrane biomimetic polymer nanomedicine in tumor cells. Figure 8 Figure c is the distribution of doxorubicin in the cell membrane biomimetic polymer nanomedicine in tumor cells. Figure 8 Middle (d) is the fluorescence distribution of the entire tumor cells.

[0070] Fig. 9 This is a graph showing the cytotoxicity of the cell membrane biomimetic polymer nanomedicine constructed by the present invention to mouse 4T1 tumor cells. DETAILED DESCRIPTION

[0071] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. 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 creative work belong to the scope of protection of the present invention.

[0072] In a first aspect, some embodiments of the present invention provide a cell membrane biomimetic polymer nanomedicine, wherein the nanomedicine comprises a cell membrane and polymer nanoparticles, wherein the polymer nanoparticles are composed of a polymer, positively charged metal peroxide nanoparticles and a chemotherapeutic drug; The structural formula of the polymer is as follows: ; The polyethyleneimine in the polymer is polyethyleneimine with a weight average molecular weight of 1000-10000, and n=3-30; the polyethylene glycol in the polymer is polyethylene glycol with a weight average molecular weight of 2000-10000, and m=50-250.

[0073] In the polymer obtained by the present invention, the grafting rate of phenylboronic acid structure monomer is 10%-80%.

[0074] In some embodiments, the metal peroxide includes one or more of calcium peroxide, copper peroxide, iron oxide and manganese dioxide.

[0075] In some embodiments, the chemotherapy drug includes one or more of doxorubicin, camptothecin, paclitaxel and gemcitabine.

[0076] In some embodiments, the method for preparing the positively charged metal peroxide nanoparticles comprises: The metal dichloride, ammonia water and pentalysine are dissolved in an ethanol solution, and a hydrogen peroxide solution is added, reacted, and centrifuged to obtain a solid precipitate; the solid matter is then placed in a saturated carbon dioxide environment to obtain positively charged metal peroxide nanoparticles.

[0077] In the present invention, the solid material is placed in a saturated carbon dioxide environment, and calcium carbonate is deposited on the surface of the metal peroxide. The calcium carbonate can improve the stability of the cell membrane biomimetic polymer nanomedicine during the storage process.

[0078] In some embodiments, the metal dichloride includes one or more of calcium chloride, cupric chloride, ferrous chloride, and manganese chloride.

[0079] In some embodiments, the ethanol solution is prepared by mixing anhydrous ethanol with deionized water, wherein the volume ratio of anhydrous ethanol to deionized water is (5-29):1.

[0080] In some embodiments, the mass concentration of the metal dichloride in the ethanol solution is 1%-30%.

[0081] In some embodiments, the mass concentration of the hydrogen peroxide solution is 3%.

[0082] In some embodiments, the ratio of the metal dichloride to pentalysine is (10-100):1 by mass.

[0083] In some embodiments, the ratio of the aqueous ammonia solution, the ethanol solution, and the hydrogen peroxide solution is 1:(100-1000):2 by volume.

[0084] In some embodiments, the reaction time is 1 min-20 min.

[0085] In some embodiments, the standing time is 1.5h-4h.

[0086] In some embodiments, the cell membrane is a tumor cell.

[0087] In a second aspect, some embodiments of the present invention also provide a method for preparing the cell membrane biomimetic polymer nanomedicine, comprising the following steps: The polymer and positively charged metal peroxide nanoparticles are dissolved and dispersed in deionized water, and an organic solvent containing a chemotherapeutic drug is added, dialyzed, and freeze-dried to obtain polymer nanoparticles; the cell membrane and the polymer nanoparticles are mixed, and centrifuged and ultrafiltered through a micro-extruder with a polycarbonate membrane to obtain a cell membrane biomimetic polymer nanomedicine.

[0088] In some embodiments, the organic solvent includes one or more of dimethyl sulfoxide, methanol and dimethylformamide.

[0089] In some embodiments, the mass concentration of the chemotherapy drug in the organic solvent is 0.5 mg / mL-5 mg / mL.

[0090] In some embodiments, the specific operation of the dialysis includes: placing the target object in a dialysis bag, and then placing the dialysis bag in deionized water for dialysis.

[0091] In some embodiments, the dialysis time is 2h-6h.

[0092] In some embodiments, the ratio of the polymer to the positively charged metal peroxide nanoparticles is (3-30):1 by mass.

[0093] In some embodiments, the mass concentration of the polymer in deionized water after being dissolved and dispersed is 3 mg / mL-30 mg / mL.

[0094] In some embodiments, the ratio of the dissolved and dispersed deionized water to the organic solvent containing the chemotherapeutic drug is (10-100):1 by volume.

[0095] In some embodiments, the ratio of the cell membrane to the polymer nanoparticles is 1:(10-50) by mass.

[0096] In some embodiments, the specific process of passing through the micro-extruder with a polycarbonate membrane is to sequentially pass through a micro-extruder with a polycarbonate membrane with a pore size of 400 nm and a polycarbonate membrane with a pore size of 200 nm.

[0097] In some embodiments, the method for preparing the polymer includes: Step S1: dissolving 16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)hexadecanoic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mixing well, adding polyethylene glycol-polyethyleneimine and N,N-diisopropylethylamine, reacting, dialyzing, and freeze-drying to obtain compound a; Step S2: dissolving 4-carboxyphenylboronic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mixing evenly, adding compound a and N,N-diisopropylethylamine, reacting, adding diethylamine, dialyzing, and freeze-drying to obtain compound b; Step S3: dissolve 3-(diethylamino)propionic acid hydrochloride, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mix well, add compound b and N,N-diisopropylethylamine, dialyze, and freeze-dry to obtain a polymer.

[0098] In some embodiments, in step S1, the polyethylene glycol in the polyethylene glycol-polyethylene imine is polyethylene glycol with a weight average molecular weight of 2000-10000, and the polyethylene imine is polyethylene imine with a weight average molecular weight of 1000-10000.

[0099] In some embodiments, in step S1, the ratio of 16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)hexadecanoic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is (1-2):(1-2):1 by mass.

[0100] In some embodiments, in step S1, before uniform mixing, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(1000-10000) by volume.

[0101] In some embodiments, in step S1, the mass concentration of 1-hydroxybenzotriazole in dimethylformamide is 0.5 mg / mL-5 mg / mL.

[0102] In some embodiments, in step S1, there are multiple ways to achieve uniform mixing. In the present invention, stirring is used. As an example, the stirring time is 20 minutes.

[0103] In some embodiments, in step S1, the mass ratio of the polyethylene glycol-polyethyleneimine to 1-hydroxybenzotriazole is (10-100):1.

[0104] In some embodiments, in step S1, the volume ratio of N,N-diisopropylethylamine added before and after mixing is (1-2):(1-5).

[0105] In some embodiments, in step S1, the reaction time is 1 h-3 h.

[0106] In some embodiments, in step S1, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days.

[0107] In some embodiments, in the mixture of dimethylformamide and deionized water, the volume ratio of dimethylformamide to deionized water is 1:(100-1000).

[0108] In some embodiments, in step S2, the ratio of 4-carboxyphenylboric acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is 1:(2-4):(2-4) by mass.

[0109] In some embodiments, in step S2, before mixing, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(50-200) by volume.

[0110] In some embodiments, in step S2, the mass concentration of 4-carboxyphenylboronic acid in dimethylformamide is 3 mg / mL-10 mg / mL.

[0111] In some embodiments, in step S2, there are multiple ways to mix evenly. In the present invention, stirring is used. As an example, the stirring time is 20 minutes.

[0112] In some embodiments, in step S2, the volume ratio of N,N-diisopropylethylamine added before and after mixing is 1:1.

[0113] In some embodiments, in step S2, the mass concentration of compound a in dimethylformamide is 0.1 mg / mL-1 mg / mL.

[0114] In some embodiments, in step S2, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days.

[0115] In some embodiments, in the mixture of dimethylformamide and deionized water, the volume ratio of dimethylformamide to deionized water is 1:(100-1000).

[0116] In some embodiments, in step S3, the ratio of 3-(diethylamino)propionic acid hydrochloride, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is 1:(2-10):(2-10) by mass.

[0117] In some embodiments, in step S3, before uniform mixing, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(1000-10000) by volume.

[0118] In some embodiments, in step S3, the mass concentration of 3-(diethylamino)propionic acid hydrochloride in dimethylformamide is 0.05 mg / mL-1 mg / mL.

[0119] In some embodiments, in step S3, there are multiple ways to mix evenly. In the present invention, stirring is used. As an example, the stirring time is 20 minutes.

[0120] In some embodiments, in step S3, the volume ratio of N,N-diisopropylethylamine added before and after mixing is (1-2):(1-2).

[0121] In some embodiments, in step S3, the mass concentration of compound b in dimethylformamide is 1 mg / mL-10 mg / mL.

[0122] In some embodiments, in step S3, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days.

[0123] In some embodiments, in the mixture of dimethylformamide and deionized water, the volume ratio of dimethylformamide to deionized water is 1:(100-1000).

[0124] In a third aspect, some embodiments of the present invention also provide applications of the cell membrane biomimetic polymer nanomedicine in the treatment of cancer.

[0125] The following is described in conjunction with specific embodiments: Embodiment 1: (1) Synthesis of polymers: 16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)hexadecanoic acid (70 mg), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (60 mg), 1-hydroxybenzotriazole (50 mg) and N,N-diisopropylethylamine (35 μL) were dissolved in dimethylformamide (50 mL). After stirring for 20 minutes, polyethylene glycol-polyethyleneimine (PEG-PEI, PEG molecular weight of 5000, PEI molecular weight of 2000; 1 g) and N,N-diisopropylethylamine (35 μL) were added. After reacting for 2 hours, the mixture was dialyzed in a mixed solution of dimethylformamide and deionized water (wherein the volume ratio of dimethylformamide to deionized water was 1:100) for 2 days, and then dialyzed in deionized water for 2 days, and freeze-dried to obtain compound a. 4-Carboxyphenylboronic acid (300 mg), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (720 mg), 1-hydroxybenzotriazole (600 mg) and N,N-diisopropylethylamine (420 μL) were dissolved in dimethylformamide (50 mL), stirred for 20 minutes, compound a (25 mg) and N,N-diisopropylethylamine (420 μL) were added, and diethylamine (50 mL) was added after stirring for 2 hours. The mixture was dialyzed in a mixture of dimethylformamide and deionized water (wherein the volume ratio of dimethylformamide to deionized water was 1:100) for 2 days, and then dialyzed in deionized water for 2 days. The mixture was freeze-dried to obtain compound b. 3-(Diethylamino)propionic acid hydrochloride (DA, 3 mg), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (10 mg), 1-hydroxybenzotriazole (10 mg) and N,N-diisopropylethylamine (10 μL) were dissolved in dimethylformamide (50 mL), stirred for 20 minutes, and then compound b (250 mg) and N,N-diisopropylethylamine (10 μL) were added, and dialyzed in a mixed solution of dimethylformamide and deionized water (wherein the volume ratio of dimethylformamide to deionized water was 1:100) for 2 days, and then dialyzed in deionized water for 2 days, and freeze-dried to obtain a polymer; (2) Preparation of positively charged calcium peroxide nanoparticles (CaO2): Calcium chloride (3.5 mg), ammonia water (100 μL) and pentalysine (200 μg) were dissolved in 35 mL of ethanol solution (wherein the volume ratio of anhydrous ethanol to deionized water = 29:1), and a 3% mass concentration of hydrogen peroxide solution (200 μL) was added under stirring. After stirring for 5 minutes, the mixture was centrifuged (32000 rpm, 15 min) to collect white nanoparticles. The collected nanoparticles were then placed in saturated carbon dioxide (CO2) and allowed to stand for 2 h to form a calcium carbonate protective layer on the surface, thereby obtaining positively charged calcium peroxide nanoparticles. (3) Preparation of polymer nanoparticles (Poly / CaO2 / Dox): 300 mg of the aforementioned polymer and 10 mg of positively charged calcium peroxide nanoparticles were dispersed in 10 mL of deionized water, and 1 mL of dimethyl sulfoxide solution containing doxorubicin (Dox, 5 mg) was added. The mixture was stirred for 30 min and dialyzed for 2 h. The polymer nanoparticles were then freeze-dried to obtain the polymer nanoparticles. (4) Extraction of 4T1 tumor cell membrane: The cells were disrupted by ultrasonic disruption, and the cell contents were removed by low-speed centrifugation (500-3000 g). The supernatant was purified by high-speed centrifugation (10000 g) to obtain the cell membrane precipitate. (5) Preparation of cell membrane biomimetic polymer nanomedicine (Mem-Poly / CaO2 / Dox): 20 g of isolated and extracted cell membrane was fully mixed with 1 g of polymer nanoparticles, and passed through a micro-extruder with 400 nm and 200 nm polycarbonate membranes in turn. Finally, the excess fused cell membrane was removed by centrifugal ultrafiltration to obtain cell membrane biomimetic polymer nanomedicine.

[0126] Comparative Example 1: Preparation of positively charged calcium peroxide nanoparticles (CaO2-CO2) without deposited calcium carbonate: Calcium chloride (3.5 mg), ammonia water (100 μL) and pentalysine (200 μg) were dissolved in 35 mL of ethanol solution (wherein the volume ratio of anhydrous ethanol to deionized water = 29:1), and a 3% mass concentration of hydrogen peroxide solution (200 μL) was added under stirring. After stirring for 5 minutes, the white nanoparticles were collected by centrifugation (32000 rpm, 15 min).

[0127] Comparative Example 2: Polymer-calcium peroxide nanoparticles (Poly / CaO2): 300 mg of polymer and 10 mg of positively charged calcium peroxide nanoparticles were dispersed in 10 mL of deionized water, stirred for 30 min, dialyzed for 2 hours, and freeze-dried to obtain polymer-calcium peroxide nanoparticles.

[0128] Comparative Example 3: Doxorubicin containing cell membrane biomimetic polymer (Mem-Poly / Dox): The separated and extracted cell membrane was fully mixed with doxorubicin, and passed through a micro-extruder with 400nm and 200nm polycarbonate membranes in turn, and finally the excess fused cell membrane was removed by centrifugal ultrafiltration.

[0129] Results and Analysis: The morphological characteristics of the positively charged metal peroxide nanoparticles (calcium peroxide nanoparticles), polymer nanoparticles and cell membrane biomimetic polymer nanomedicine in Example 1 were imaged using a transmission electron microscope. Each sample was spotted on a copper grid at a concentration of 1 mg / mL, negatively stained with 2% phosphotungstic acid, and observed using a transmission electron microscope after the sample was naturally dried in the air. Figure 1-Figure 3As shown, calcium peroxide nanoparticles, polymer nanoparticles and cell membrane biomimetic polymer nanodrugs all have spherical structures and are evenly distributed. A dynamic light scattering instrument was used to measure the hydrated particle size and Zeta potential of calcium peroxide nanoparticles, polymer nanoparticles and cell membrane biomimetic polymer nanodrugs. The laser was incident at 90° and the incident wavelength was 633nm. Each sample was measured in parallel 3 times, the equilibrium time was set to 2 minutes, and the measurement temperature was 25°C. The concentration of each measured sample was 1mg / mL. The hydrated particle size distribution range of calcium peroxide nanoparticles is 1nm-10nm, and the hydrated particle size distribution range of polymer nanoparticles and cell membrane biomimetic polymer nanodrugs is 10nm-300nm. In addition, due to the external wrapping of the cell membrane, the hydrated particle size of the cell membrane biomimetic polymer nanodrug is slightly larger than that of the polymer nanoparticles. As shown Figure 4 As shown, since polylysine is adsorbed on the surface of calcium peroxide nanoparticles, they are positively charged. After calcium carbonate is deposited on their surface, the potential of the calcium peroxide nanoparticles decreases slightly because the positively charged groups on the surface are covered. Since the polymer has a large negative charge, after the calcium peroxide nanoparticles self-assemble with the polymer and doxorubicin to form polymer nanodrugs, the charge is reversed and the Zeta potential changes from positive to negative. After coating the tumor cell membrane, the Zeta potential of the cell membrane biomimetic polymer nanodrug further decreases because the cell membrane is negatively charged, which is beneficial for it to maintain stability during circulation in the body.

[0130] Take appropriate amounts of doxorubicin, polymer-calcium peroxide nanoparticles (Poly / CaO2) and cell membrane biomimetic polymer nanomedicine, dissolve them in dimethyl sulfoxide (DMSO), prepare sample solutions of appropriate concentrations, and scan the UV absorption spectrum in the wavelength range of 300nm-800nm. The results are as follows Figure 5 As shown, the characteristic peak of doxorubicin appears at 490nm, the characteristic peak of polymer-calcium peroxide nanoparticles appears at 350nm, and the calcium peroxide nanoparticle cell membrane biomimetic polymer nanodrug also has absorption peaks at 490nm and 350nm, confirming that doxorubicin and polymer-calcium peroxide nanoparticles are successfully loaded in the cell membrane biomimetic polymer nanodrug.

[0131] CD44 protein and E-cadherin protein are 4T1 tumor cell membrane surface antigens. The expression of these two proteins in 4T1 tumor cell membrane (4T1 Membrane) and cell membrane biomimetic polymer nanomedicine was detected by Western blotting. Figure 6 It can be seen that CD44 protein and E-cadherin protein are expressed in the cell membrane biomimetic polymer nanomedicine, indicating that the nanomedicine retains the antigen structure on the surface of the tumor cell membrane, and the homologous targeting of the tumor cell membrane is retained in the preparation. The nanomedicine obtained by the present invention can accurately act on the target tumor cells.

[0132] The in vitro release of drug-loaded nanoparticles was evaluated by dialysis method, and the release conditions were as follows: the release medium was PBS buffer with a mass fraction of 0.1% Tween 80, the temperature was 37°C, and the shaker speed was 100 rpm. 1 mL of cell membrane biomimetic polymer nanodrug was put into a dialysis bag (MWCO=10000), and 1 mL of release medium was taken out at the predetermined time (0h, 1h, 2h, 4h, 6h, 12h, 24h, 48h), and fresh blank medium was added. The content of doxorubicin was determined by fluorescence spectrophotometry, and the fluorescence intensity of the sample under excitation at a wavelength of 488nm and emission at a wavelength of 580nm was determined, and the in vitro release curve of doxorubicin in cell membrane biomimetic polymer nanodrug was drawn according to the test results. Figure 7 As shown in the figure, the cell membrane biomimetic polymer nanodrug is relatively stable under systemic circulation conditions (pH=7.4), with a slow drug release rate, reaching a platform at about 20%; under weakly acidic tumor microenvironment conditions, due to the mutual repulsion of protonated hydrophilic end groups of the polymer, the release rate of doxorubicin increases slightly, reaching a platform at about 30%; under the stimulation of weakly acidic tumor microenvironment (pH 6.5) and intratumoral ATP concentration (1μM), doxorubicin is rapidly released from the cell membrane biomimetic polymer nanodrug, indicating that the nanodrug can achieve controllable drug release in response to changes in the tumor microenvironment.

[0133] Alexa Fluor™ 647-labeled calcium peroxide nanoparticles were used to prepare cell membrane biomimetic polymer nanomedicines, which were co-incubated with 4T1 breast cancer cells for 2 h. The cell nuclei were stained with Hoechst 33342 and the cells were observed using confocal laser scanning microscopy (CLSM). Figure 8 As shown in the figure, calcium peroxide adheres to the tumor cell membrane, while the fluorescence of doxorubicin (Dox) is evenly distributed in the cytoplasm and cell membrane at the same time, while doxorubicin that enters cells through endocytosis is generally distributed in the lysosomes of the cytoplasm. This phenomenon shows that cell membrane biomimetic polymer nanomedicine can release calcium peroxide nanoparticles, which create small holes on the surface of tumor cells, promote doxorubicin to enter tumor cells directly through the transmembrane holes, and avoid the degradation of drugs by lysosomes.

[0134] 4T1 cells in the logarithmic growth phase and in good growth condition were digested, resuspended and counted, and the cells were counted at 5×10 3The cells were inoculated into a 96-well plate at a density of 100 μL / well. After the cells adhered to the wall, the original culture medium was aspirated and replaced with a serum-free culture medium containing cell membrane biomimetic polymer doxorubicin (Mem-Poly / Dox) and cell membrane biomimetic polymer nanodrug (Mem-Poly / CaO2 / Dox) (calcium concentration was 2 μg / mL, doxorubicin concentration was 5 μg / mL). The cell-free drug group was used as the control group, and the cell-free drug group was used as the zero adjustment group. Each group had 6 replicate wells and was incubated for 48 hours. After the incubation, 20 μL of CCK8 solution was added to each well. After incubation for another 2 hours, the absorbance (OD) was detected at 490 nm using an ELISA reader, and the cell viability (CellViability) was calculated according to the following formula.

[0135] Cell survival rate = (OD experimental group - OD zero adjustment group) / (OD control group - OD zero adjustment group) × 100%; The results are as follows Fig. 9 As shown, the cell membrane biomimetic polymer doxorubicin can produce limited killing effect on 4T1 cells, and the introduction of calcium peroxide nanoparticles significantly enhances the tumor cell killing effect of doxorubicin (Dox). This is because the cell membrane biomimetic polymer nanomedicine prepared by the present invention can intelligently respond to changes in the tumor microenvironment to release calcium peroxide nanoparticles and cause tumor cell membrane perforation, promote doxorubicin to directly enter the tumor cells through the transmembrane pores, avoid lysosome degradation of the drug, and increase the effective drug concentration of doxorubicin in tumor cells.

[0136] It should be noted that the terms used in the present invention are only for describing specific embodiments, rather than limiting the scope of the present application. As shown in the present specification, unless the context clearly indicates an exception, the words "one", "a", "a kind of" and / or "the" do not specifically refer to the singular, but may also include the plural. The terms "include", "comprise" or any other variant thereof are intended to cover non-exclusive inclusion, so that the process, method or device including a series of elements includes not only those elements, but also includes other elements not explicitly listed, or also includes elements inherent to such process, method or device. In the absence of more restrictions, the elements defined by the sentence "include one..." do not exclude the presence of other identical elements in the process, method or device including the elements.

[0137] It should also be noted that the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. Unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", etc. should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be an indirect connection through an intermediate medium, or it can be a connection between the two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0138] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the technical solutions of the embodiments of the present invention.

Claims

1. A cell membrane biomimetic polymer nanomedicine, characterized in that: A cell membrane biomimetic polymer nanomedicine, the nanomedicine comprises a cell membrane and polymer nanoparticles, the polymer nanoparticles are composed of a polymer, positively charged metal peroxide nanoparticles and a chemotherapeutic drug; The structural formula of the polymer is as follows: ； The polyethyleneimine in the polymer is polyethyleneimine with a weight average molecular weight of 1000-10000, and n=3-30; the polyethylene glycol in the polymer is polyethylene glycol with a weight average molecular weight of 2000-10000, and m=50-250.

2. The nano drug according to claim 1, characterized in that The metal peroxide includes one or more of calcium peroxide, copper peroxide, iron oxide and manganese dioxide; The chemotherapy drugs include one or more of doxorubicin, camptothecin, paclitaxel and gemcitabine; The cell membrane is a tumor cell.

3. The nano drug according to claim 1, characterized in that The preparation method of the positively charged metal peroxide nanoparticles comprises: The metal dichloride, ammonia water and pentalysine are dissolved in an ethanol solution, and a hydrogen peroxide solution is added, reacted, and centrifuged to obtain a solid precipitate; the solid matter is then placed in a saturated carbon dioxide environment to obtain positively charged metal peroxide nanoparticles.

4. The nano drug according to claim 3, characterized in that The metal dichloride includes one or more of calcium chloride, cupric chloride, ferrous chloride, and manganese chloride; The mass concentration of the metal dichloride in the ethanol solution is 1%-30%; The mass concentration of the hydrogen peroxide solution is 3%; The ratio of the metal dichloride to pentalysine is (10-100):1 by mass; In terms of volume, the ratio of the aqueous ammonia, the ethanol solution, and the hydrogen peroxide solution is 1:(100-1000):2; The reaction time is 1min-20min; The standing time is 1.5h-4h.

5. The method for preparing the cell membrane biomimetic polymer nanomedicine according to any one of claims 1 to 4, characterized in that: The following steps are involved: The polymer and positively charged metal peroxide nanoparticles are dissolved and dispersed in deionized water, and an organic solvent containing a chemotherapeutic drug is added, dialyzed, and freeze-dried to obtain polymer nanoparticles; the cell membrane and the polymer nanoparticles are mixed, and centrifuged and ultrafiltered through a micro-extruder with a polycarbonate membrane to obtain a cell membrane biomimetic polymer nanomedicine.

6. The preparation method according to claim 5, characterized in that: The organic solvent includes one or more of dimethyl sulfoxide, methanol and dimethylformamide; The mass concentration of the chemotherapy drug in the organic solvent is 0.5 mg / mL-5 mg / mL; The dialysis time is 2h-6h; The ratio of the polymer to the positively charged metal peroxide nanoparticles is (3-30):1 by mass; The mass concentration of the polymer in deionized water after being dissolved and dispersed is 3 mg / mL-30 mg / mL; The ratio of the deionized water after dissolution and dispersion to the organic solvent containing the chemotherapeutic drug is (10-100):1 by volume; The mass ratio of the cell membrane to the polymer nanoparticles is 1:10-50.

7. The preparation method according to claim 5, characterized in that: The preparation method of the polymer comprises: Step S1: dissolving 16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)hexadecanoic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mixing well, adding polyethylene glycol-polyethyleneimine and N,N-diisopropylethylamine, reacting, dialyzing, and freeze-drying to obtain compound a; Step S2: dissolving 4-carboxyphenylboronic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mixing evenly, adding compound a and N,N-diisopropylethylamine, reacting, adding diethylamine, dialyzing, and freeze-drying to obtain compound b; Step S3: dissolve 3-(diethylamino)propionic acid hydrochloride, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole and N,N-diisopropylethylamine in dimethylformamide, mix well, add compound b and N,N-diisopropylethylamine, dialyze, and freeze-dry to obtain a polymer.

8. The preparation method according to claim 7, characterized in that: In the step S1, the polyethylene glycol in the polyethylene glycol-polyethylene imine is polyethylene glycol with a weight average molecular weight of 2000-10000, and the polyethylene imine is polyethylene imine with a weight average molecular weight of 1000-10000; In the step S1, the ratio of 16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)hexadecanoic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is (1-2):(1-2):1 by mass; In the step S1, before being uniformly mixed, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(1000-10000) by volume; In the step S1, the mass concentration of 1-hydroxybenzotriazole in dimethylformamide is 0.5 mg / mL-5 mg / mL; In the step S1, the mass ratio of the polyethylene glycol-polyethyleneimine to 1-hydroxybenzotriazole is (10-100):

1. In the step S1, the volume ratio of N,N-diisopropylethylamine added before and after mixing is (1-2):(1-5); In the step S1, the reaction time is 1h-3h; In step S1, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days.

9. The preparation method according to claim 7, characterized in that: In the step S2, the ratio of 4-carboxyphenylboronic acid, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is 1:(2-4):(2-4) by mass; In the step S2, before being uniformly mixed, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(50-200) by volume; In the step S2, the mass concentration of 4-carboxyphenylboronic acid in dimethylformamide is 3 mg / mL-10 mg / mL; In the step S2, the volume ratio of N,N-diisopropylethylamine added before and after mixing is 1:1; In the step S2, the mass concentration of compound a in dimethylformamide is 0.1 mg / mL-1 mg / mL; In step S2, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days; In step S3, the ratio of 3-(diethylamino)propionic acid hydrochloride, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 1-hydroxybenzotriazole is 1:(2-10):(2-10) by mass; In the step S3, before being uniformly mixed, the ratio of N,N-diisopropylethylamine to dimethylformamide is 1:(1000-10000) by volume; In step S3, the mass concentration of 3-(diethylamino)propionic acid hydrochloride in dimethylformamide is 0.05 mg / mL-1 mg / mL; In the step S3, the volume ratio of N,N-diisopropylethylamine added before and after mixing is (1-2):(1-2); In the step S3, the mass concentration of the compound b in dimethylformamide is 1 mg / mL-10 mg / mL; In step S3, the specific steps of performing dialysis include: dialysis in a mixture of dimethylformamide and deionized water for 2 days, and then dialysis in deionized water for 2 days.

10. Use of the cell membrane biomimetic polymer nanomedicine according to any one of claims 1 to 4 or the cell membrane biomimetic polymer nanomedicine obtained by the preparation method according to any one of claims 5 to 9 in the preparation of drugs for treating cancer.

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