Polymers inducing tumor immunogenic cell death, methods of making and uses

Abstract

The application belongs to the technical field of polymer medicine synthesis, and particularly relates to a polymer for inducing tumor immunogenic cell death, a preparation method and application. The polymer for inducing tumor immunogenic cell death provided by the application has a structure as shown in formula I: in formula I, m is an integer of 110-120; n is an integer of 70-75; p is an integer of 21-79; and R is one of a tertiary amine group, a sulfide group and a tertiary amine sulfide group. The polymer provided by the application has pH responsiveness, can be self-assembled to obtain a polymer nanovesicle, can load drugs, can induce tumor immunogenic cell death (ICD) after application to form an in-situ tumor vaccine, can realize controllable release of a hydrophobic immunoadjuvant, can polarize tumor-associated macrophages (TAMs), and is suitable for tumor immunotherapy application.

Description

Technical Field

[0001] This invention belongs to the field of polymer drug synthesis technology, specifically relating to polymers that induce tumor immunogenic cell death, their preparation methods, and applications. Background Technology

[0002] Immunogenic cell death (ICD) refers to the transformation of tumor cells from a non-immunogenic state to an immunogenic state after exposure to external stimuli, stimulated by the host's immune response. Typical immunological features of ICD include damage-associated molecular patterns (DAMPs), such as the secretion of calreticulin (CRT), high-mobility group box 1 (HMGB1), adenosine triphosphate (ATP), and the release of tumor-associated antigens. These DAMPs promote dendritic cell (DC) maturation and migration, presenting antigens to T cells and activating the host immune system. Constructing nanocarriers that can directly induce ICD simplifies the composition of nanoplatforms and, due to their own immune function, effectively promotes therapeutic efficacy, making them significant for tumor treatment.

[0003] In situ tumor vaccines utilize antigens released directly from dying tumor cells, overcoming the problem of tumor antigen heterogeneity. By binding immune adjuvants to tumor-associated antigens (TAAs), in situ tumor vaccine formation can be promoted. The most significant advantage of this process is that it eliminates the need for additional antigens, relying instead on the tumor's own antigen library generated after external stimulation, thus saving time and costs. By loading cRGD cyclic peptide-modified nanoparticles with immune adjuvants (such as rethimod R848), they can selectively target tumor cells with high αvβ3 expression on their cell surface (such as melanoma cells B16F10), promoting in situ cancer vaccine formation by enhancing tumor cell internalization.

[0004] Immunosuppressive factors in the tumor microenvironment (TME), such as tumor-associated macrophages (TAMs), significantly hinder immunotherapeutic efficacy. TAMs primarily exist in the immunosuppressive M2 form, accounting for 50% of all immune cells in tumor tissue, thus facilitating tumor evasion of immune surveillance. M1-type TAMs are pro-inflammatory and can eliminate tumors by presenting antigens to T lymphocytes. Therefore, polarizing M2-TAMs to the M1 form using polarizing agents (such as R848) is one method for modulating TAMs to remodel the TME.

[0005] Mannose-modified nanoparticles can target tumor markers (TAMs) and enhance polarization by increasing intracellular uptake. Furthermore, tumor cells highly express differentiation cluster 47 (CD47) on their surface, which participates in tumor resistance to macrophage phagocytosis via the CD47 / SIRPα signaling pathway. Therefore, both TAM polarization and CD47 / SIRPα signaling pathway blockade are essential for remodeling the tumor microenvironment (TME).

[0006] Therefore, how to simplify the platform composition by constructing a polymer nanoplatform that can induce ICD on its own, while loading drugs with immune adjuvant function and TAM polarization ability, and modifying the formulation surface with targeting ligands that target tumor cells and TAMs respectively, can be used to construct tumor vaccines in situ in vivo and regulate TME, achieving the dual purpose of coordinating natural and adaptive immunity to enhance therapeutic efficacy. Summary of the Invention

[0007] To overcome the shortcomings of existing technologies, the primary objective of this invention is to provide a polymer that induces tumor immunogenic cell death. This polymer is pH-responsive, capable of self-assembling into polymer nanovesicles, and can be loaded with drugs. Upon application, it can induce tumor ICDs to form in situ tumor vaccines and achieve controlled release of hydrophobic immune adjuvants, polarizing tumor-associated macrophages (TAMs), making it suitable for tumor immunotherapy.

[0008] A second objective of the present invention is to provide a method for preparing the polymer that induces tumor immunogenic cell death.

[0009] A third objective of this invention is to provide the application of the polymer described above that induces tumor immunogenic cell death.

[0010] The fourth objective of this invention is to provide an anti-tumor nanomedicine that, upon application, can induce tumor ICDs to form in situ tumor vaccines and achieve controlled release of hydrophobic immune adjuvants, polarizing tumor-associated macrophages (TAMs), making it suitable for tumor immunotherapy applications.

[0011] One of the objectives of this invention is achieved through the following technical solution:

[0012] A polymer that induces tumor immunogenic cell death, the structure of which is shown in Formula I:

[0013]

[0014] In Formula I, m is an integer from 110 to 120; n is an integer from 70 to 75; and p is an integer from 21 to 79.

[0015] R is a tertiary amine group. sulfide group tertiary amine sulfide group One of them.

[0016] The polymer for inducing tumor immunogenic cell death provided by this invention is an ABC-type block polymer. Block A is a polyethylene glycol (PEG) segment, with the corresponding repeating unit m being an integer from 110 to 120; block B is a polymethyl methacrylate (PMMA) segment, with the corresponding repeating unit n being an integer from 70 to 75; and block C is a composite segment (P(PPMA-MPA-DEA)) with the corresponding repeating unit p being an integer from 21 to 79. This polymer has a novel chemical structure, can induce tumor immunogenic cell death (ICD), and is responsive to pH. Specifically, at pH 7.4, the molecular weight of the hydrophobic portion of the polymer is 1.5 times or more that of the hydrophilic portion (block A), making it highly suitable for the preparation of polymer nanovesicles (a supramolecular aggregate of amphiphilic molecules self-assembling). Furthermore, the PPMA groups in the polymer can induce tumor immunogenic cell death by grafting active groups. Furthermore, polymer nanovesicles prepared by self-assembly of the polymers of the present invention can be further connected with different target heads to achieve targeting of different cells. Hydrophobic drug adjuvants can also be further encapsulated in the hydrophobic core of the vesicles, thereby not only enriching the functions of the polymer, but also enhancing the anti-tumor effect of the drug.

[0017] To optimize the polymer's immunofunctional structure and application performance, as a preferred embodiment, m is 114, n is 73, and p is 79; R is a tertiary amine thioether group.

[0018] The second objective of this invention is achieved by the following technical solution:

[0019] A method for preparing a polymer that induces tumor immunogenic cell death includes the following steps:

[0020] (1) Polyethylene glycol-amino is subjected to an amidation reaction with 4-cyano-4-(phenylthiocarbonylthio)pentanoic acid N-succinimide ester to obtain polyethylene glycol-4-cyano-4-(phenylthiocarbonylthio)pentanoamide, denoted as PEG-CPPA.

[0021] (2) PEG-CPPA was polymerized with methyl methacrylate to obtain product A, denoted as PEG-PMMA;

[0022] (3) Reaction of PEG-PMMA with N-propargylmethacrylamide yields product B, denoted as PEG-PMMA-PPPMA;

[0023] (4) PEG-PMMA-PPPMA was reacted with mercaptopropionic acid to obtain product E, which was denoted as PEG-PMMA-P (PPMA-MPA);

[0024] (5) PEG-PMMA-P (PPMA-MPA) is esterified with diethylaminoethanol to obtain product F, which is the polymer that induces tumor immunogenic cell death, denoted as PEG-PMMA-P (PPMA-MPA-DEA).

[0025] The polymer preparation method of this invention uses PEG-CPAA as a chain transfer agent to polymerize methyl methacrylate monomer via living RAFT, obtaining a polymer with controllable molecular weight and narrow molecular weight distribution. Further, after polymerizing N-propynemylated methacrylamide via living RAFT, a click reaction is carried out with mercaptopropionic acid, followed by an esterification reaction with diethylaminoethanol, yielding a polymer with controllable molecular weight and narrow molecular weight distribution capable of inducing ICD. This process is simple to operate, has mild process conditions, and facilitates the efficient preparation of polymers.

[0026] As a preferred embodiment, in step (1), the molar ratio of the reaction between polyethylene glycol-amino and 4-cyano-4-(phenylthiocarbonylthio)valerate N-succinimide ester is 2:3; in step (2), the molar ratio of the reaction between PEG-CPPA and methyl methacrylate is 1:100; in step (3), the molar ratio of the reaction between PEG-PMMA and N-propyne methacrylamide is 1:25; in step (4), the molar ratio of the reaction between PEG-PMMA-PPPMA and mercaptopropionic acid is 1:50; and in step (5), the molar ratio of the reaction between PEG-PMMA-P (PPMA-MPA) and diethylaminoethanol is 1:100.

[0027] The third objective of this invention is achieved by the following technical solution:

[0028] The above-mentioned polymers that induce tumor immunogenic cell death are used in the preparation of antitumor nanomedicines.

[0029] The fourth objective of this invention is achieved by the following technical solution:

[0030] An anti-tumor nanomedicine, the preparation method of which includes the following steps: self-assembling the polymer that induces tumor immunogenic cell death to obtain polymer nanovesicles, then modifying the polymer nanovesicles with a target and loading them with drugs that have immune adjuvant function and polarizing macrophage function, thereby obtaining the anti-tumor nanomedicine.

[0031] As a preferred embodiment, the self-assembly process is as follows: the polymer that induces tumor immunogenic cell death is dissolved in tetrahydrofuran, and then phosphate buffer is added to prepare uniformly distributed polymer nanovesicles by solvent exchange.

[0032] As a preferred embodiment, the target is mannose and / or cRGD cyclic peptide. cRGD cyclic peptide, also known as C(RGDFK) cyclic peptide, is a five-membered cyclic peptide composed of arginine, glycine, aspartic acid, D-phenylalanine, and lysine, with a ring formed by amide bonds at the beginning and end. It is a melanoma-targeting peptide that binds to α-cells on the surface of tumor cells. v β3 receptor binding can serve as a tool for peptide targeting in certain types of cancer. Mannose is an active substance that can target TAMs, increasing polarization rate by enhancing intracellular uptake.

[0033] As a preferred option, the drug with immune adjuvant function and macrophage polarization function is rethimod R848.

[0034] As a preferred embodiment, polymer nanovesicles are modified with a target and loaded with drugs possessing both immunoadjuvant and macrophage-polarizing functions. Specifically, the polymer nanovesicles are uniformly mixed with a polymer containing a target and a drug possessing both immunoadjuvant and macrophage-polarizing functions, and then the antitumor nanomedicine is prepared by solvent displacement. Further, the polymer containing the target is prepared by esterifying mannose and / or cRGD cyclic peptides to the carboxyl terminus of polyethylene glycol.

[0035] This invention does not specifically limit the types of tumor cells targeted by the anti-tumor nanomedicines; in specific applications, adaptive designs can be made for different tumor cells. As a preferred embodiment, the anti-tumor nanomedicine is an anti-melanoma drug.

[0036] The antitumor nanomedicine provided by this invention is prepared using a polymer that induces tumor immunogenic cell death. The antitumor nanomedicine prepared by this invention is a pH-responsive nanomedicine, maintaining stability at pH 7.4. After entering the cell, it first enters the lysosome, escapes from the lysosome, and then targets the mitochondria. The nanocarrier self-regulates metabolic oxidative phosphorylation and GSDMD upregulates pyroptosis, directly inducing tumor ICD. The drug is released at pH 5.0. Specifically, nanoparticles modified with a target that target tumor cells release a drug that can act as an immune adjuvant, activating dendritic cells (DCs) to form an in situ tumor vaccine and activating T cells. Nanoparticles modified with a target that target macrophages release a drug that can polarize macrophages and activate the immune system. Animal experiments in melanoma have confirmed that the antitumor nanomedicine of this invention significantly prolongs the median survival of mice and induces significant tumor cell death. Therefore, the polymer that induces tumor immunogenic cell death and the prepared antitumor nanomedicine of this invention can be effectively applied to tumor immunotherapy, providing a new therapeutic strategy for effectively treating cancer by coordinating adaptive and innate immune responses in time and space using immunoactive nanomaterials.

[0037] Compared with the prior art, the comprehensive beneficial effects of the present invention are as follows:

[0038] The polymer for inducing tumor immunogenic cell death provided by this invention can induce immunogenic death in various tumor cells, expose tumor-associated antigens, and activate in vivo immune responses. Furthermore, the polymer can self-assemble into immunologically active nanoparticles, effectively simplifying the nanoplatform. In addition, the polymer for inducing tumor immunogenic cell death provided by this invention can be linked to different targets to achieve targeting of different cells, thereby reducing the toxicity of nanomedicines to normal cells and immune cells, achieving reduced toxicity and enhanced efficacy. Simultaneously, the polymer for inducing tumor immunogenic cell death provided by this invention can self-assemble into vesicles, and the hydrophobic core of the vesicles can be loaded with hydrophobic drugs. These hydrophobic drugs can be immunoadjuvants that polarize macrophages, thus giving a single polymer multiple functions, making it highly suitable for tumor immunotherapy applications, and showing broad application value in cancer immunotherapy, especially in the immunotherapy of melanoma. Attached Figure Description

[0039] Figure 1 The above are the 1H NMR spectrum and structural diagram of the polymer PEG-CPPA involved in Example 2 of this invention.

[0040] Figure 2 The above are the 1H NMR spectrum and structural diagram of the polymer PEG-PMMA involved in Example 2 of this invention.

[0041] Figure 3The above are the 1H NMR spectrum and structural diagram of the polymer PEG-PMMA-PDEA involved in Example 2 of this invention.

[0042] Figure 4 The 1H NMR spectrum and structural diagram of the polymer PEG-PMMA-PPPMA involved in Example 2 of this invention are shown below.

[0043] Figure 5 The 1H NMR spectrum and structural diagram of the polymer PEG-PMMA-P (PPMA-ME) involved in Example 2 of this invention are shown below.

[0044] Figure 6 The 1H NMR spectrum and structural diagram of the polymer PEG-PMMA-P (PPMA-MPA) involved in Example 2 of this invention are shown below.

[0045] Figure 7 The 1H NMR spectrum and structural diagram of the polymer PEG-PMMA-P (PPMA-MPA-DEA) involved in Example 2 of this invention are shown below.

[0046] Figure 8 The particle size distribution diagrams are shown for the three polymer nanovesicles prepared in Example 3 of this invention.

[0047] Figure 9 The image shows the results of ICD induced by the three polymer nanovesicles in Experiment Example 1; among them, Figure 9 A shows the results of CRT exposure on the cell surface; Figure 9 B represents the HMGB1 content in the cell supernatant; Figure 9 C represents the ATP content in the cell supernatant;

[0048] Figure 10 This is a graph showing the in vitro release of R848 under different conditions within 24 hours in Experiment Example 2;

[0049] Figure 11 This is a graph showing the cell uptake detected by flow cytometry in Experiment Example 3;

[0050] Figure 12 For example 4, cRGD-pRNC Thioether+DEA Image showing the results of DC maturation induced by @R848;

[0051] Figure 13 Man-pRNC in Experimental Example 5 Thioether+DEA The result of @R848-induced polarization in RAW264.7 cells;

[0052] Figure 14 cRGD-pRNC in Experimental Example 6 Thioether+DEA mix Man-pRNC Thioether+DEATargeting results in mice;

[0053] Figure 15 cRGD-pRNC in Experimental Example 7 Thioether+DEA mix Man-pRNC Thioether+DEA The graph shows the in vivo antitumor activity of @R848 in the B16-F10 tumor model and its effect on prolonging survival when used in combination with antibodies; among which... Figure 15 A shows the changes in tumor volume in mice after treatment. Figure 15 B is a graph showing the changes in mouse body weight. Figure 15 C shows the mouse body weight after combination with the antibody. Figure 15 D is the mouse survival curve after combination with antibody. Detailed Implementation

[0054] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. It should be understood that the following embodiments are merely further illustrative of the present invention and not intended to limit it. Unless otherwise specified, the raw materials and reagents used in the following embodiments are commercially available. In the following embodiments, the cRGD cyclic peptide involved is from Shanghai Taopu Biotechnology Co., Ltd.

[0055] Example 1

[0056] This embodiment provides a polymer that induces tumor immunogenic cell death, with a structure as shown in Formula I:

[0057]

[0058] Where m is 114; n is 73; p is 79; and R is a tertiary amine sulfide group.

[0059] Example 2

[0060] This embodiment describes the preparation of the polymer (PEG-PMMA-P(PPMA-MPA-DEA)) that induces tumor immunogenic cell death as described in Example 1, and also describes the preparation of polymers PEG-PMMA-PDEA and PEG-PMMA-P(PPMA-ME).

[0061] The specific method includes the following steps:

[0062] I. Synthesis of PEG-PMMA-PDEA

[0063] The synthetic route of PEG-PMMA-PDEA is shown in route ①.

[0064] Route ①:

[0065]

[0066] The specific steps are as follows:

[0067] (1) Synthesis of macromolecular reversible addition fracture transfer radical polymerization (RAFT) reagents

[0068] RAFT reagent, or polyethylene glycol-4-cyano-4-(phenylthiocarbonylthio)pentanamide (PEG-CPPA), is obtained by amidation reaction of polyethylene glycol-amino (PEG-NH2, purchased from Biochempeg, molecular weight 5000, polyvinyl alcohol repeating unit number 114) with N-succinimide 4-cyano-4-(phenylthiocarbonylthio)pentanolic acid (CPPA, TCI (Shanghai) Technology Co., Ltd.).

[0069] The specific process of the amidation reaction was as follows: Under nitrogen atmosphere and an ice-water bath, a 2 mL solution of CPPA (56.4 mg, 0.15 mmol) in dichloromethane was added dropwise to a solution of PEG-NH2 (500 mg, 0.10 mmol) and triethylamine (0.01 mmol, 1.02 mg), and the reaction was carried out at room temperature for 24 h. The product was precipitated with ice-cold ether, filtered, and dried under vacuum to obtain the RAFT reagent PEG-CPPA.

[0070] Figure 1 The 1H NMR spectrum of the PEG-CPPA prepared in this step and the structural diagram of the compound are shown. Figure 1 1H NMR spectrum ( 1 The characterization results by H NMR were as follows: PEG: δ 3.63, CPPA: δ 7.38, 7.57, 7.90. These results indicate that the bonding rate of CPPA in the PEG-CPPA mixture is 98%.

[0071] (2) Synthesis of PEG-PMMA-PDEA

[0072] The macromolecular triblock copolymer polyethylene glycol-polymethyl methacrylate-polydiethylaminoethyl methacrylate (PEG-PMMA-PDEA) was obtained by polymerization with RAFT reagent.

[0073] The specific process is as follows: Under nitrogen atmosphere, PEG-CPPA (100 mg, 0.02 mmol), methyl methacrylate (MMA, 200 mg, 2.00 mmol), and azobisisobutyronitrile (AIBN, 0.492 mg, 0.003 mmol) were dissolved in 1,4-dioxane (2 mL). After stirring at 65 °C for 24 h, the product was precipitated with ice-cold diethyl ether, filtered, and dried under vacuum to obtain the diblock product A, denoted as PEG-PMMA.

[0074] Figure 2The image shows the 1H NMR spectrum of PEG-PMMA and the structural diagram of the compound. Figure 2 The characterization results are as follows: 1 HNMR (400MHz, CDCl3): PEG: δ3.63; PMMA: δ3.65. Depend on Figure 2 It can be seen that in the structural formula of PEG-PMMA, m=114, n=100. The molecular weight of PEG-PMMA is 5.0-10.0 kg / mol, that is, PEG(5.0k)-PMMA(10.0k).

[0075] Further, under nitrogen atmosphere, the diblock PEG-PMMA (64.5 mg, 0.0043 mmol) prepared in the previous step, diethylaminoethyl methacrylate (DEA, 19.96 mg, 0.108 mmol), and azobisisobutyronitrile (AIBN, 0.106 mg, 0.0006 mmol) were dissolved in 1,4-dioxane and reacted under sealed conditions for 48 h. After the reaction, the mixture was precipitated with ice-cold diethyl ether, then filtered and dried under vacuum to obtain product C, denoted as PEG-PMMA-PDEA.

[0076] Figure 3 The image shows the 1H NMR spectrum of PEG-PMMA-PDEA and the structural diagram of the compound. Figure 3 The characterization results are as follows: 1 H NMR (400MHz, CDCl3): PEG: δ 3.63; PDEA: δ 0.86, 1.26. (From...) Figure 3 The characterization results show that in the PEG-PMMA-PDEA structure, m = 114, n = 100, and p = 21. The molecular weight of PEG-PMMA-PDEA is 5.0-10.0-3.8 kg / mol, i.e., PEG(5.0k)-PMMA(10.0k)-PDEA(3.8k).

[0077] II. Synthesis of PEG-PMMA-P (PPMA-ME)

[0078] The synthetic route for PEG-PMMA-P (PPMA-ME) is shown in route ②.

[0079] Route 2:

[0080]

[0081] The specific steps are as follows: N-propynylmethacrylamide (PPMA) can be obtained by amidation reaction. Specifically, 4.68 g (44.8 mmol) of methacrylamide chloride, 2.057 g (37.4 mmol) of propargylamine, and 463.6 mg (3.8 mmol) of 4-dimethylaminopyridine were weighed. The 4-dimethylaminopyridine (DMAP, 463.6 mg, 3.8 mmol) was dissolved in DCM, and the propargylamine (2.057 g, 37.4 mmol) was diluted with 12 mL of DCM. The DMAP solution and methacrylamide chloride (4.68 g, 44.8 mmol) were mixed thoroughly, and then added dropwise to the propargylamine solution under ice-water bath and nitrogen atmosphere. After the addition was complete, the mixture was allowed to return to room temperature, and the reaction was carried out at room temperature for 24 h. After the reaction was complete, the solution was purified by silica gel column chromatography (mobile phase ratio of n-hexane / ethyl acetate = 5 / 1) to obtain a pale yellow oily liquid PPMA with a yield of 25%.

[0082] The diblock product A obtained above, namely PEG-PMMA (100 mg, 0.0067 mmol), PPMA (34 mg, 0.276 mmol), and AIBN (0.164 mg, 0.001 mmol), was dissolved in 1,4-dioxane and reacted under sealed conditions for 48 h. After the reaction, the mixture was precipitated with ice-cold diethyl ether, then filtered and dried under vacuum to obtain the triblock product B, denoted as PEG-PMMA-PPPMA.

[0083] Figure 4 The image shows the 1H NMR spectrum of PEG-PMMA-PPPMA and the structural diagram of the compound. Figure 4 The characterization results are as follows: 1 ¹H NMR (400MHz, CDCl₃): PEG: δ 3.63; PMMA: δ 3.65; PPPMA: δ 1.81. From this characterization result, the PEG-PMMA-PPPMA structure has m = 114, n = 100, and p = 51. The molecular weight of PEG-PMMA-PPPMA is 5.0-10.0-6.3 kg / mol, i.e., PEG(5.0kJ)-PMMA(10.0kJ)-PPPMA(6.3kJ).

[0084] Further, PEG-PMMA-P (PPMA-ME) was obtained by a click reaction of PEG-PMMA-PPPMA with mercaptoethanol. In short, PEG-PMMA-PPPMA (41 mg, 0.00195 mmol), mercaptoethanol (7.59 mg, 0.0973 mmol), and dimethyl benzoate (DMPA, 6.23 mg, 0.0243 mmol) were dissolved in DMF. Subsequently, the solution was added sequentially to a closed UV reactor and irradiated with UV light for 2 hours (350 nm). After the reaction was complete, the product was precipitated with ice-cold diethyl ether, filtered, and dried under vacuum to obtain product D, denoted as PEG-PMMA-P (PPMA-ME).

[0085] Figure 5 The image shows the 1H NMR spectrum of PEG-PMMA-P (PPMA-ME) and the structural diagram of the compound. Figure 5 The characterization results are as follows: 1 ¹H NMR (400MHz, CDCl₃): PEG: δ 3.63; PMMA: δ 3.65; P(PPMA-ME): δ 2.89, 2.97. From this characterization result, the structural formula of PEG-PMMA-P(PPMA-ME) shows m = 114, n = 100, and p = 51. The molecular weight of PEG-PMMA-P(PPMA-ME) is 5.0-10.0-10.4 kg / mol, i.e., PEG(5.0k)-PMMA(10.0k)-P(PPMA-ME)(10.4k).

[0086] III. Synthesis of PEG-PMMA-P (PPMA-MPA-DEA)

[0087] The synthetic route for PEG-PMMA-P (PPMA-MPA-DEA) is shown in route ② above.

[0088] The specific steps are as follows: Triblock PEG-PMMA-PPPMA (154 mg, 0.0070 mmol), mercaptopropionic acid (MPA) (37.4 mg, 0.353 mmol), and dimethyl benzoate (DMPA, 22.60 mg, 0.088 mmol) were dissolved in DMF and irradiated with ultraviolet light (350 nm) for 2 hours under sealed conditions. After the reaction was completed, the product was precipitated with ice-cold diethyl ether, filtered, and dried under vacuum to obtain product E, denoted as PEG-PMMA-P (PPMA-MPA).

[0089] Figure 6 The image shows the 1H NMR spectrum of PEG-PMMA-P (PPMA-MPA) and the structural diagram of the compound. Figure 6 The characterization results are as follows: 1¹H NMR (400MHz, CDCl₃): PEG: δ 3.63; PMMA: δ 3.65; P(PPMA-MPA): δ 2.89, 2.97. From this characterization result, the structure of PEG-PMMA-P(PPMA-MPA) shows m = 114, n = 73, and p = 79. The molecular weight of PEG-PMMA-P(PPMA-MPA) is 5.0-7.3-14.5 kg / mol, i.e., PEG(5.0k)-PMMA(7.3k)-P(PPMA-MPA)(14.5k).

[0090] PEG-PMMA-P (PPMA-MPA) and diethylaminoethanol were esterified to obtain PEG-PMMA-P (PPMA-MPA-DEA). Specifically, PEG-PMMA-P (PPMA-MPA) (112.5 mg, 0.0042 mmol), N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC·HCl, 30.19 mg, 0.157 mmol), and 4-dimethylaminopyridine (DMAP, 12.8 mg, 0.105 mmol) were dissolved in DMF. First, EDC and DMAP were added to PEG-PMMA-P (PPMA-MPA), and nitrogen gas was introduced for 30 min. Then, diethylaminoethanol (49.2 mg, 0.42 mmol) was added, and nitrogen gas was introduced for another 15 min. The mixture was sealed and reacted at room temperature for 24 h. After the reaction was completed, the product was dialyzed with ultrapure water for 24 hours and then freeze-dried to obtain product F, denoted as PEG-PMMA-P (PPMA-MPA-DEA). This compound F is the polymer that induces tumor immunogenic cell death provided in Example 1 of this invention.

[0091] Figure 7 The image shows the 1H NMR spectrum of PEG-PMMA-P (PPMA-MPA-DEA) and the structural diagram of the corresponding compound. Figure 7 The characterization results are as follows: 1 ¹H NMR (400MHz, CDCl₃): PEG: δ 3.63; PMMA: δ 3.65; P(PPMA-MPA-DEA): δ 0.86, 1.25. From this characterization result, the structure of PEG-PMMA-P(PPMA-MPA-DEA) shows m = 114, n = 73, and p = 79. The molecular weight of PEG-PMMA-P(PPMA-MPA-DEA) is 5.0-7.3-18.4 kg / mol, i.e., PEG(5.0kJ)-PMMA(7.3kJ)-P(PPMA-MPA-DEA(18.4kJ).

[0092] Example 3

[0093] This example demonstrates the application of the tumor immunogenic cell death-inducing polymer (PEG-PMMA-P(PPMA-MPA-DEA)) prepared in Example 2 in the preparation of antitumor nanomedicines. Antitumor nanomedicines are prepared by self-assembling the tumor immunogenic cell death-inducing polymer into polymer nanovesicles, then modifying the polymer nanovesicles with a target and loading them with drugs possessing immune adjuvant and macrophage polarization functions. The target is mannitol or cRGD. The drug with immune adjuvant and macrophage polarization functions is retimote R848.

[0094] The specific preparation method is as follows:

[0095] I. Synthesis of cRGD-PEG-PMMA-PPPMA and Man-PEG-PMMA-PPPMA

[0096] The synthesis route is shown in route ③.

[0097] Route 3:

[0098]

[0099] The specific steps are as follows:

[0100] Man-PEG-PMMA-PPPMA was obtained by amidation reaction of COOH-PEG-PMMA-PPPMA with amino-modified mannose (Man-NH2). The structural formula of COOH-PEG-PMMA-PPPMA is shown in route diagram ③, and its molecular weight was also determined by... 1 HNMR characterization. Specifically, to obtain Man-PEG-PMMA-PPPMA, NHS (0.489 mg, 0.0043 mmol) and EDC·HCl (0.815 mg, 0.0043 mmol) were first added to a mixed solution of COOH-PEG-PMMA-PPPMA (35 mg, 0.0028 mmol) and triethylamine (TEA), and stirred for 1.5 h under N2 protection to obtain NHS-PEG-PMMA-PPPMA. NHS-PEG-PMMA-PPPMA was then added dropwise to a Man-NH2 solution containing TEA under an ice-water bath and N2 atmosphere. The reaction was then placed in a 30°C water bath and stirred in the dark for 24 h. The product was dialyzed and lyophilized to obtain Man-PEG-PMMA-PPPMA.

[0101] The synthesis of cRGD-PEG-PMMA-PPPMA is similar to that of Man-PEG-PMMA-PPPMA, except that cRGD-NH2 is substituted for Man-NH2. The prepared cRGD-PEG-PMMA-PPPMA and Man-PEG-PMMA-PPPMA were analyzed by nuclear magnetic resonance. 1 H NMR characterization and verification of the structure 1 H NMR (400MHz, CDCl3): PEG: δ3.63; cRGD: δ1.84; Man: δ2.28).

[0102] II. Preparation of Polymer Nanovesicles

[0103] Three polymer nanovesicles were prepared using a solvent exchange method. Specifically, the polymers PEG-PMMA-PDEA, PEG-PMMA-P (PPMA-ME), and PEG-PMMA-P (PPMA-MPA-DEA) prepared in Example 2 were dissolved in tetrahydrofuran (100 μL, 5 mg / mL) and slowly added to phosphate buffer (PB, 900 μL, pH 7.4) to obtain three uniformly distributed polymer nanovesicles, which were designated pRNC. DEA NC Thioether ,pRNC Thioether+DEA The particle size distribution and transmission electron microscopy images of the three polymer nanovesicles are shown below. Figure 8 As shown.

[0104] Combination Figure 8 The results show that pRNC DEA The hydrodynamic dimensions are 126.5 ± 3.6 nm, with a distribution of 0.22 ± 0.012; NC Thioether The hydrodynamic dimensions are 132.8 ± 5.6 nm, with a distribution of 0.22 ± 0.025; pRNC Thioether+DEA The hydrodynamic size of the polymer is 140.7 ± 3.5 nm, and the distribution is 0.129 ± 0.010. This indicates that the above polymers can all be obtained as uniformly distributed nanoparticles through solvent exchange.

[0105] III. cRGD-pRNC Thioether+DEA @R848 and Man-pRNC Thioether+DEA Preparation of @R848

[0106] Two types of drug-loaded polymer nanovesicles, cRGD-pRNC, were prepared using a solvent exchange method. Thioether+DEA @R848 and Man-pRNC Thioether+DEA@R848. The preparation method is as follows: PEG-PMMA-P (PPMA-MPA-DEA), cRGD-PEG-PMMA-PPPMA, and Man-PEG-PMMA-PPPMA prepared in step one are dissolved in tetrahydrofuran (5 mg / mL). Then, PEG-PMMA-P (PPMA-MPA-DEA) (80 μL), cRGD-PEG-PMMA-PPPMA or Man-PEG-PMMA-PPPMA (20 μL), and rethimod R848 (10 mg / mL, 5 μL) are mixed and slowly added to phosphate buffer (PBS, 900 μL, pH 7.4) to obtain uniformly distributed drug-loaded polymer nanovesicles, denoted as cRGD-pRNC. Thioether+DEA @R848、Man-pRNC Thioether+DEA @R848.

[0107] Experimental Example 1 pRNC DEA NC Thioether pRNC Thioether+DEA Assay for mediating immunogenic cell death

[0108] The polymer nanovesicles pRNC prepared in Example 3 of this invention DEA NC Thioether pRNC Thioether+DEA and NC MMA NC yne The ability of polymer nanocarriers to mediate immunogenic cell death in tumor cells was assessed. Among them, NC... MMA Referring to step two in Example 3, the polymer nanocarrier prepared from PEG-PMMA via solvent exchange was used; NC yne Referring to step two in Example 3, the polymer nanocarrier prepared from PEG-PMMA-PPPMA by solvent exchange method.

[0109] The ability of tumor cells to mediate immunogenic death was characterized by detecting CRT exposure, ATP release, and HMGB1 release. For characterizing CRT exposure: B16F10 cells were seeded in six-well plates and cultured overnight, then NC cells were harvested. MMA pRNC DEA NC yne NC Thioether and pRNC Thioether+DEA (Concentration of 100 μg / mL) were added to each well and incubated for 48 h. After washing with PBS (×3), digestion, and centrifugation, the solution was used with Alexa. 488 anti-CRT staining, washing with PBS, and centrifugation were performed. Cells were then resuspended in PBS (0.5 mL per sample), stained with PI, incubated at room temperature for 10 min, and analyzed by flow cytometry. HMGB1 release characterization: Cells were cultured overnight in 24-well plates, then NC was added. MMA pRNC DEA NC yne NC Thioether and pRNC Thioether+DEA (All concentrations were 100 μg / mL). After treatment for 48 h, the cells were washed with PBS (×3), digested, and centrifuged. Cells were then processed using Alexa... 488 anti HMGB1 staining (200×, 40 min), washed with PBS, fixed with 4% paraformaldehyde, Alexa staining. 647 anti-HMGB1 staining and nuclear staining. Cell images were acquired using laser confocal microscopy. Supernatants from different treatments were collected, and HMGB1 release was detected using ELISA. ATP release assay: ATP secretion from B16F10 cells after different treatments was detected using the ATP Assay Kit (S0026). B16F10 cells were seeded on six-well plates and cultured overnight. MMA pRNC DEA NC yne NC Thioether and pRNC Thioether+DEA Add the solution to each well and incubate for 24 h. Add 30 μL of cell culture supernatant to 70 μL of ATP assay working solution. Measure the RLU value using enzyme-linked immunosorbent assay (ELISA) within 30 min. Measure the total protein concentration in the six-well plate using a BCA kit. Compare the ATP release per unit protein concentration in each group.

[0110] The results are as follows Figure 9 As shown

[0111] Figure 9 The results showed that the cells were treated with pRNC DEA NC Thioether pRNC Thioether+DEA All treatments mediated increased exposure of CRT on the surface of tumor cells, with pRNC showing the most significant improvement. Thioether+DEA The CRT signal was strongest. In the cell supernatant, the content of HMGB1 increased, and pRNC... Thioether+DEA The group that released the most HMGB1 also released the most ATP. Based on the above results, it can be concluded that the polymer nanovesicles pRNC... Thioether+DEA Compared to nanovesicle pRNC DEA and NC Thioether Its CRT exposure, ATP release, and HMGB1 release effects are all the strongest, therefore this invention selects pRNCThioether+DEA It will be used as the target polymer for subsequent experimental studies.

[0112] Experimental Example 2 cRGD-pRNC Thioether+DEA @R848 and Man-pRNC Thioether+DEA Characterization and release test of @R848

[0113] The drug-loaded nanoparticles cRGD-pRNC prepared in Example 3 of this invention Thioether+DEA @R848 and Man-pRNC Thioether+DEA @R848 was characterized and its in vitro release was studied. To obtain the leak conditions, cRGD-pRNC was separately... Thioether+DEA @R848 and Man-pRNC Thioether+DEA The R848 (0.5 mL) release bag was placed in 25 mL of release medium and then placed in a shaker (37℃, 200 rpm). At pre-set time points, 5 mL of release medium was removed and the same volume of fresh medium was added. The release media were PBS (pH 7.4, 10 mM, 150 mM NaCl) and acetate / sodium acetate buffer (pH 5.0, 10 mM, 150 mM NaCl). The release amount of R848 at different time points was determined using a fluorescence spectrophotometer. The results are as follows. Figure 10 As shown.

[0114] Depend on Figure 10 In vitro drug release behavior indicates that cRGD-pRNC Thioether+DEA @R848、Man-pRNC Thioether+DEA In acetate buffer (pH 5.0, 10 mM, 150 mM NaCl), the cumulative release of R848 within 24 h reached 61% and 60%, respectively, while in PBS (pH 7.4, 10 mM, 150 mM NaCl), the cumulative release was only 23% and 24%. These results indicate that cRGD(Man)-pRNC Thioether+DEA @R848 has good pH sensitivity.

[0115] Experimental Example 3 cRGD-pRNC Thioether+DEA @R848 and Man-pRNC Thioether+DEA @R848's in vitro targeting

[0116] cRGD-pRNC was detected using flow cytometry and CLSM. Thioether+DEA The targeting of the cells was investigated. FITC was used instead of R848 as a fluorescence model encapsulated in nanovesicles. B16-F10 cells (mouse skin melanoma cells) were seeded in six-well plates and cultured overnight using flow cytometry, followed by the addition of free FITC and cRGD-pRNC. Thioether+DEA@FITC and pRNC Thioether+DEA @FITC. After incubation for 10 min, 30 min, and 1 h, cells were digested with trypsin, centrifuged, resuspended in PBS, and analyzed by flow cytometry.

[0117] Using cRGD-pRNC Thioether+DEA Man-pRNC was prepared using a similar solvent exchange method. Thioether+DEA It was characterized using DLS and TEM. (Compared with cRGD-pRNC) Thioether+DEA Similarly, targeting was assessed using flow cytometry, with RAW264.7 cells (macrophages) replacing B16F10 cells. Results were as follows... Figure 11 As shown.

[0118] Figure 11 The results showed that, compared with pRNC Thioether+DEA Compared to @FITC, cRGD-pRNC Thioether+DEA @FITC-treated B16F10 cells showed more pronounced fluorescence shifts, indicating that cRGD has good targeting properties. Man-pRNC Thioether+DEA @FITC treatment resulted in a fluorescence shift in RAW264.7 cells compared to pRNC. Thioether+DEA The @FITC group showed stronger targeting, demonstrating significant Man's targeting ability. Therefore, cRGD and Man modifications enhanced targeting of B16F10 and RAW264.7 cells, respectively.

[0119] Experimental Example 4: cRGD-pRNC Thioether+DEA @R848 induces DC cell maturation

[0120] DC maturation was assessed by flow cytometry. B16-F10 cells were seeded in six-well plates and cultured overnight. PBS and NC were then added. MMA cRGD-pRNC Thioether+DEA cRGD-pRNC Thioether+DEA @R848、cRGD-pRNC Thioether+DEA @R848 mixMan-pRNC Thioether+DEA @R848 (concentration 100 μg / mL) was added to each well. DC cells were seeded in six-well plates overnight. After 48 h of incubation with the drug, 1 mL of cell supernatant was mixed with 1 mL of fresh culture medium and added to the DC cells. After 24 h of culture, the cells were washed with PBS (×3), digested, and centrifuged. They were then stained with APC-antiCD86 and Percp cy5.5-antiCD80, washed with PBS, and centrifuged again. Finally, the cells were resuspended in PBS (0.5 mL per sample) and analyzed by flow cytometry. Results are as follows: Figure 12 As shown.

[0121] Figure 12 The results showed that B16-F10 cells, after undergoing cRGD-pRNC treatment... Thioether+DEA The DAMP released after treatment can effectively mature DC cells.

[0122] Experimental Example 5: Man-pRNC Thioether+DEA @R848 induces macrophage polarization

[0123] Macrophage polarization is primarily characterized by cell surface markers via flow cytometry. RAW 264.7 cells were seeded in 12-well plates and cultured overnight. IL-4 (10 ng / mL) and recombinant mouse macrophage colony-stimulating factor 1 (MCS-F1, 10 ng / mL) were added to polarize M0 cells to the M2 phenotype. PBS and Man-pRNC were then added. Thioether+DEA R848, Man-pRNC Thioether+DEA @R848, cRGD-pRNC Thioether+DEA @R848 mix Man-pRNC Thioether+DEA @R848 was added to M2 phenotype RAW264.7 (R848: 1 μg / mL) and incubated for 4 h. The original culture medium was aspirated, and fresh culture medium was added and incubated for 20 h. After washing with PBS, trypsin digestion, and centrifugation, cells were stained with Percp cy5.5-anti CD80, FITC-anti CD206, and PE-anti F4 / 80 (40 min), resuspended in PBS, and analyzed by flow cytometry. Cells treated with LPS (10 ng / mL) and IFN-γ (10 ng / mL) polarized M0 to the M1 phenotype, serving as a positive control and a CD80 single-staining group. Results are as follows: Figure 13 As shown.

[0124] Figure 13 The results showed that Man-pRNC Thioether+DEA @R848 treatment increased CD80 expression and decreased CD206 expression on the cell surface. Therefore, Man-pRNC Thioether+DEA @R848 can effectively polarize M2 RAW264.7 cells to the M1 phenotype.

[0125] Experimental Example 6: cRGD-pRNC Thioether+DEA @R848 and Man-pRNC Thioether+DEA @R848 tumor enrichment and in vivo targeting

[0126] In in vivo tumor accumulation studies, lipophilic cell membrane fluorescent probes DID (1,1'-bis(octadecyl-3,3,3',3'-tetramethylindole-dicarbocyanine perchlorate) and DIR (1,1-octadecyl-3,3,3,3-tetramethylindolecyanine iodide) were used to replace R848 as fluorescent models and were respectively coated onto cRGD-pRNC. Thioether+DEA and Man-pRNC Thioether+DEA In this study, LLC cells (small cell lung cancer cells) were used instead of B16-F10 cells to inoculate the backs of mice. When the tumor volume reached 150 mm², [the tumor was successfully treated]. 3 When left and right, inject free DID (G1), free DID mix DIR (G2), and pRNC into the tail vein respectively. Thioether+DEA @DID(G3), cRGD-pRNC Thioether+DEA @DID(G4) and cRGD-pRNC Thioether+DEA @DID mix Man-pRNC Thioether+DEA @DIR(G5) (n=3). Fluorescence intensity of tumor tissue was detected by IVIS imaging system at 2, 4, 8, 12, and 24 h post-injection. 24 h later, tumor tissue was digested with collagenase IV (50 U / mL) in 1640 medium containing 2% FBS for 2 hours (37°C). The supernatant was filtered through a 70 μm filter and centrifuged (1200 rpm, 5 min), and cells were collected from the bottom. Cells were stained with FITC antiPD-L1, washed with PBS, centrifuged, resuspended in PBS, and detected by flow cytometry. The targeting of TAM was similar to that of the tumor tissue described above, using free DIR (G1′), free DIR mix DID (G2), and pRNC. Thioether+DEA @DIR(G3′), Man-pRNC Thioether+DEA @DID(G4′) and cRGD-pRNC Thioether+DEA @DID mix Man-pRNCThioether+DEA@DIR(G5) alternative formulation, with PE antiF4 / 80 as the antibody replacement. Results are as follows. Figure 14 As shown.

[0127] Figure 14 The results showed that after intravenous injection, the in vivo fluorescence intensity of cRGD and Man nanoformations initially increased and then decreased within 24 hours, reaching its maximum at 8 hours. The in vivo fluorescence intensity of the cRGD and Man modified groups was higher than that of pRNC. Thioether+DEA @DID indicates cRGD-pRNC Thioether+DEA and Man-pRNC Thioether+DEA It exhibits good in vivo targeting. Further, based on in vitro results, cRGD-pRNCT showed good targeting in tumor cells. hioether+DEA @DID and cRGD-pRNCThioether+DEA @DID mix Man-pRNC Thioether+DEA The proportion of @DIR is relatively high in tumor-associated macrophages with Man-pRNC. Thioether+DEA @DIR、cRGD-pRNC Thioether+DEA @DID mix Man-pRNC Thioether+DEA The proportion of @DIR is relatively high. This indicates that cRGD-pRNC Thioether+DEA Man-pRNC can effectively target tumor cells in vivo. Thioether+DEA It can effectively target TAM in vivo.

[0128] Experimental Example 7 cRGD-pRNC Thioether+DEA @R848 mix Man-pRNC Thioether+DEA Antitumor activity of @R848 in melanoma models and prolongation of survival when used in combination with antibodies

[0129] A melanoma model was established by inoculating the right side of C57BL / 6 mice with B16-F10 cells. When the tumor grew to approximately 50 mm... 3 Mice were injected via tail vein on days 6, 9, and 12 post-inoculation, and tumor size and body weight were observed and recorded. Mice treated with antibodies (anti-CD47 and anti-PD-1) were subcutaneously injected with B16-F10 cells; tumors reached 120 mm² in size. 3 The nanoformulation was administered via tail vein injection on days 8, 11, and 14, and the antibody was administered intravenously on days 9, 12, and 15. Mice survival was observed and recorded. Experimental results are as follows: Figure 15 As shown.

[0130] Figure 15 The results show that cRGD-pRNC Thioether+DEA @R848 mix Man-pRNC Thioether+DEA @R848 treatment significantly inhibited tumor growth in mice. Further combination with antibody therapy significantly prolonged median survival in mice.

[0131] Based on the above experimental results, the polymer providing this invention, which induces tumor immunogenic cell death, can self-assemble into a pH-responsive polymer nanoplatform. This nanoplatform itself can induce the ICD effect in melanoma B16-F10 cells. Further target modification of this nanoplatform and loading of drugs with immune adjuvant and macrophage polarization functions to construct a composite nanoplatform yields novel antitumor nanomedicines. Thioether+DEA / R848 mix Man-pRNC Thioether+DEA / R848, this drug can target tumor cells to form an in situ cancer vaccine and target TAMs for polarization, thereby achieving spatiotemporal coordinated activation of adaptive and innate immune responses, demonstrating the advantage of this platform in utilizing a single polymer backbone for multiple functional combinations. Furthermore, animal model experiments with melanoma have confirmed that the anti-tumor nanomedicine provided by this invention can effectively enhance the anti-tumor effect of drugs and prolong the median survival of animals, indicating that this drug has the potential to exert immune checkpoint blockade effects. Therefore, this invention provides a new strategy for forming multifunctional nanocarriers using immunofunctional polymers, possessing powerful cancer immunomodulatory effects. Moreover, the polymers and anti-tumor nanomedicines of this invention, due to their simple synthesis ideas and design processes, have strong potential for clinical translation and application, and have broad application prospects in the immunotherapy of melanoma.

[0132] The above embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention shall fall within the scope of protection claimed by the present invention.

Claims

1. A polymer inducing immunogenic cell death of a tumor, characterized in that, The structure of the polymer inducing tumor immunogenic cell death is shown as formula I: In formula I, m is an integer of 110-120; n is an integer of 70-75; p is an integer of 21-79; R is a tertiary amine group sulfide groups tertiary amine sulfide groups one of the following 2. The polymer inducing tumor immunogenic cell death according to claim 1, characterized in that, m is 114, n is 73, p is 79; R is a tertiary amine sulfide group 3. A method for preparing a polymer inducing immunogenic cell death of a tumor according to claim 1, characterized by, The method comprises the following steps: (1) Amide reaction of polyethylene glycol-amino and 4-cyano-4-(phenylthiocarbonylthio) pentanoic acid N-succinimidyl ester to obtain polyethylene glycol-4-cyano-4-(phenylthiocarbonylthio) pentanamide, denoted as PEG-CPPA; (2) Polymerization reaction of PEG-CPPA and methyl methacrylate to obtain product A, denoted as PEG-PMMA; (3) Reaction of PEG-PMMA and N-propargyl methacrylamide to obtain product B, denoted as PEG-PMMA-PPPMA; (4) Click reaction of PEG-PMMA-PPPMA and mercaptopropionic acid to obtain product E, denoted as PEG-PMMA-P(PPMA-MPA); (5) Esterification reaction of PEG-PMMA-P(PPMA-MPA) and diethylaminoethanol to obtain product F, which is the polymer inducing tumor immunogenic cell death, denoted as PEG-PMMA-P(PPMA-MPA-DEA).

4. The method for preparing the polymer that induces tumor immunogenic cell death according to claim 3, characterized in that, In step (1), the reaction molar ratio of polyethylene glycol-amino and 4-cyano-4-(phenylthiocarbonylthio) pentanoic acid N-succinimidyl ester is 2:3; in step (2), the reaction molar ratio of PEG-CPPA and methyl methacrylate is 1:100; in step (3), the reaction molar ratio of PEG-PMMA and N-propargyl methacrylamide is 1:25; in step (4), the reaction molar ratio of PEG-PMMA-PPPMA and mercaptopropionic acid is 1:50; and in step (5), the reaction molar ratio of PEG-PMMA-P(PPMA-MPA) and diethylaminoethanol is 1:

100.

5. Use of a polymer inducing immunogenic cell death of a tumor according to claim 1 or 2, characterized in that, The polymer inducing tumor immunogenic cell death is used in the preparation of an anti-tumor nano-drug.

6. An antitumor nano-drug, characterized in that, The preparation method of the anti-tumor nano-drug comprises the following steps: self-assembly of the polymer inducing tumor immunogenic cell death to obtain a polymer nano-vesicle, target head modification of the polymer nano-vesicle, and loading of a drug having the functions of an immune adjuvant and polarized macrophages, so that the anti-tumor nano-drug is obtained. 7.The anti-tumor nano-drug according to claim 6, characterized in that, The self-assembly process is as follows: the polymer inducing tumor immunogenic cell death is dissolved in tetrahydrofuran, then phosphate buffer is added, and a uniformly distributed polymer nano-vesicle is prepared by a solvent exchange method. 8.The anti-tumor nano-drug of claim 6, characterized in that, The target head is mannose and / or cRGD cyclic peptide. 9.The anti-tumor nano-drug of claim 6, characterized in that, The drug having the functions of an immune adjuvant and polarized macrophages is resiquimod R848.

10. The antitumor nanomedicine according to any one of claims 6 to 9, characterized in that, The anti-tumor nano-drug is an anti-melanoma drug.

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