Polymer capable of being assembled with proteins and hydrophobic small molecule drugs and preparation method thereof

Abstract

The invention discloses a polymer that can be assembled with proteins and hydrophobic small molecule drugs and a preparation method thereof. Any one of an ammonium functional group-containing monomer, an amine functional group-containing monomer and an oxirane functional group-containing monomer is selected as a main material, and a diblock polymer with a cavity functional group is obtained through a secondary ATRP reaction or a RAFT reaction. For the main material of the monomer containing an oxirane functional group, the diblock polymer with a cavity functional group is grafted with an organic molecule containing an amine functional group through organic modification, and the amine radical group is cationized to obtain a polymer that can be assembled with proteins and hydrophobic small molecule drugs. The polymer that can be assembled with proteins and hydrophobic small molecule drugs prepared by the present invention is a degradable multi-block polymer, which can be combined with hydrophobic small molecule targeted drugs, macromolecular immune and targeted drugs through host-guest self-assembly, so that the existing drug-loaded microspheres can effectively load small molecule targeted drugs, macromolecular targeted drugs and macromolecular immune drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of medical materials, and in particular relates to a polymer that can be assembled with proteins and hydrophobic small molecule drugs and a preparation method thereof. Background Art

[0002] According to the "Guidelines for the Diagnosis and Treatment of Primary Liver Cancer (2022 Edition)", the first-line drugs currently recommended for the treatment of primary liver cancer are atezolizumab combined with bevacizumab, sintilimab combined with bevacizumab, donafenib, lenvatinib, sorafenib and oxaliplatin, etc. At present, drug-loaded microsphere arterial chemoembolization (D-TACE) can load chemotherapy drugs to embolize tumor blood vessels, and at the same time, it can treat tumors by sustained release of chemotherapy drugs.

[0003] At present, the first-line drugs for clinical treatment of primary liver cancer are oxaliplatin as a chemotherapy drug, and the rest of the drugs, such as sorafenib, lenvatinib, donafenib, bevacizumab, atezolizumab and sintilimab, are targeted and immune checkpoint inhibitor drugs. First- and second-line targeted drugs, due to their poor water solubility, low bioavailability, and toxic side effects, cause many harms to patients in clinical treatment. First- and second-line immune checkpoint inhibitors can cause varying degrees of immune-related adverse reactions in the patient's lungs, liver, kidneys, gastrointestinal system, endocrine system and skin. Therefore, releasing the above drugs at the tumor site through physical targeting can effectively reduce the circulation of excessive drugs in human blood and reduce toxic side effects on the one hand, and effectively improve the utilization rate of drugs acting on tumors on the other hand. At present, the clinically used drug-loaded microsphere arterial chemoembolization (D-TACE) is to combine chemotherapy drugs with embolic microspheres, and fill the drug-loaded microspheres into tumor blood vessels through embolization, so as to continuously release drugs at the tumor site. However, the hydrophobic properties of targeted drugs and the macromolecular properties of immunosuppressant drugs mean that neither of these two types of drugs can be loaded by the drug-loaded embolic microspheres currently available on the market. Summary of the invention

[0004] In view of the technical problems existing in the above-mentioned prior art, the present invention proposes a polymer that can be assembled with proteins and hydrophobic small molecule drugs and a preparation method thereof, aiming to enable the degradable multi-block polymer to be combined with hydrophobic small molecule targeted drugs or macromolecular immune and targeted drugs through host-guest self-assembly, so that the existing drug-loaded microspheres can effectively load small molecule targeted drugs, macromolecular targeted drugs and macromolecular immune drugs.

[0005] In a first aspect of the present invention, there is provided a polymer that can be assembled with proteins and hydrophobic small molecule drugs, characterized in that it has any one of the following structural formulas shown in Formula I to Formula VI:

[0006] , , , , , , wherein the R group is a functional group containing cyclodextrin or calixarene, n is the number of ammonium blocks, m is the number of cavity-containing molecular blocks, and n and m are each independently selected from an integer greater than or equal to 2.

[0007] Wherein, the R group is a functional group containing cyclodextrin or calixarene, n is the number of ammonium blocks, m is the number of cavity-containing molecular blocks, and n and m are each independently selected from an integer greater than or equal to 2.

[0008] Preferably, the ratio of n to m is in the range of (0.25-4):1.

[0009] In some embodiments of the present invention, the polymer is selected from the following structures:

[0010] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

[0011] The second aspect of the present invention provides a method for preparing the above-mentioned polymer that can be assembled with proteins and hydrophobic small molecule drugs, comprising:

[0012] Any one of an ammonium functional group-containing monomer, an amine functional group-containing monomer and an alkylene oxide functional group-containing monomer is selected as a main material, a polymer intermediate having a grafting site is obtained through a first ATRP reaction or a RAFT reaction, a main molecule polymer containing a cavity structure or a polymer containing hydroxyl groups and ester groups is grafted to the polymer intermediate through a second ATRP reaction or a RAFT reaction, and the polymer containing hydroxyl groups and ester groups is modified by a cavity-containing organic molecule, so as to finally obtain a diblock polymer with a cavity functional group;

[0013] When the main material is a monomer containing an ammonium functional group, the diblock polymer with a cavity functional group is the polymer that can be assembled with proteins and hydrophobic small molecule drugs;

[0014] When the main material is a monomer containing an amine functional group, the diblock polymer with a cavity functional group is cationized with an amine group to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs;

[0015] When the main material is a monomer containing an alkylene oxide functional group, the diblock polymer with a cavity functional group is organically modified and grafted with an organic molecule containing an amine functional group to obtain an aminated polymer, and the amine group is cationized to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0016] In some embodiments of the present invention, the ammonium functional group-containing monomer includes any one of N-(3-aminopropyl)methacrylamide hydrochloride, 2-aminoethyl methacrylate hydrochloride, allylamine hydrochloride, 2-methylallylamine hydrochloride, O-allylhydroxylamine hydrochloride, 3-butenylamine hydrochloride, 3-methylbut-2-en-1-amine hydrochloride, 2-allylaniline hydrochloride, and N-(3-aminopropyl)methacrylamide hydrochloride.

[0017] In some embodiments of the present invention, the amino functional group-containing monomer includes any one of 2-methylallylamine, acrylamide, N-(2-amino-2-oxoethyl)acrylamide, 2-buteneamide, 4-aminostilbene, 3-aminostyrene, 4-aminostyrene, methacrylamide, 2-methylallylamine, and 4-vinylbenzylamine.

[0018] In some embodiments of the present invention, the epoxy functional group-containing monomer includes any one of glycidyl methacrylate, allyl glycidyl ether, 3,4-epoxy-1-butene, (3,4-epoxycyclohexyl) methyl acrylate, 1,2-epoxy-9-decene, 1,2-epoxy-5-hexene, 1,2-epoxy-4-vinylcyclohexane, glycidyl acrylate, glycidyl methacrylate, and 3,4-epoxy-1-butene.

[0019] In some embodiments of the present invention, the molar ratio of any one of the ammonium functional group-containing monomer, the amine functional group-containing monomer, and the alkylene oxide functional group-containing monomer to the cavity functional group-containing monomer or the cavity-containing organic molecule is (0.25-4):1.

[0020] In some embodiments of the present invention, the main molecular polymer with a cavity structure includes any one of poly(propyl methacrylate)-2-hydroxy-3-methylamino-α-cyclodextrin, poly(propyl methacrylate)-2-hydroxy-3-methylamino-β-cyclodextrin, and poly(propyl methacrylate)-2-hydroxy-3-methylamino-γ-cyclodextrin;

[0021] Optionally, the preparation method of the main molecular polymer with a cavity structure, polypropyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, polypropyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin, polypropyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin, comprises:

[0022] Any one of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin is selected as a raw material of a monomer with a cavity functional group and dissolved in an alkaline solution, and a modifier tosyl chloride is added to react for several hours, and then the filtrate is collected and subjected to reduced pressure distillation and sedimentation to obtain a solid sample I;

[0023] The solid sample I and the modifier 1,3-propylenediamine are mixed and reacted, and then distilled and precipitated under reduced pressure to obtain a solid sample II;

[0024] Solid sample II and modifier glycidyl methacrylate are dissolved in an organic solvent, nitrogen is introduced, the temperature is increased, reaction is performed, and then precipitation is performed to obtain a solid sample, which is polypropyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, polypropyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin or polypropyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin corresponding to the raw material.

[0025] In some embodiments of the present invention, the polymer containing hydroxyl groups and ester groups is any one of hydroxyethyl methacrylate, allyl 4-hydroxybenzoate, 6-(allyloxycarbonylamino)-1-hexanol, 3-hexenol lactate, hydroxyethyl acrylate, 6-hydroxyhexyl methacrylate, and hydroxypropyl acrylate;

[0026] The cavity-containing organic molecule is modified from 4-sodium azosulfonate calix[4]arene or 4-sodium azosulfonate calix[6]arene;

[0027] Optionally, the method for preparing the cavity-containing organic molecule comprises: selecting 4-azosodium calix[4]arene or 4-azosodium calix[6]arene, dissolving it in an alkaline solution, adding a modifier epibromopropane, mixing, and then precipitating with ether to obtain the cavity-containing organic molecule.

[0028] In some embodiments of the present invention, the organic molecules containing amino functional groups include diethylamine, 1,3-propylenediamine, p-phenylenediamine, adipic acid diamine, N,N'-bis(2-aminoethyl)-1,3-propylenediamine, 1,3-bis(aminomethyl)cyclohexane, N,N'-bis(3-aminopropyl)ethylenediamine, 6,6'-iminodihexylamine, 1,4-butanediol bis(3-aminopropyl) ether, 2-chloro-5-methyl-1,4-phenylenediamine, 2-chloro-5-nitro-1,4 -phenylenediamine, 2-chloro-1,4-phenylenediamine, 1,3-cyclohexanediamine, 1,4-cyclohexanediamine, 2,6-diaminoanthraquinone, 6,6'-diamino-2,2'-bipyridine, 1,4-butanediamine, 3,6-diaminocarbazole, 1,10-diaminodecane, 3,3'-diaminodipropylamine, 1,12-diaminododecane, 2,7-diaminofluorene, 1,7-heptanediamine, 1,6-hexanediamine, 2,2'-diamino-N-methyldiethylamine, 3, 3'-diamino-N-methyldipropylamine, 1,2-diamino-2-methylpropane, 1,8-diaminooctane, 1,3-diaminopentane, 1,5-diaminopentane, 1,2-diaminopropane, 1,3-propylenediamine, 2-hydroxy-1.3-diaminopropane, 1,14-diamino-3,6,9,12-tetraoxatetradecane, 2,5-diaminotoluene, 1,11-diamino-3,6,9-trioxaundecane, 1,11-diaminoundecane, Any one of 4,4'-diamino-3,3'-dimethoxybiphenyl, 2,6-dibromo-1,4-phenylenediamine, 2,5-dichloro-1,4-phenylenediamine, 2,2'-diaminodiethylamine, 3,3'-dihydroxybenzidine, 2,5-dimethyl-1,4-phenylenediamine, 2-methyl-1,5-diaminopentane, 2-methyl-1,3-propylenediamine, naphthalene-2,6-diamine, 2-nitro-1,4-phenylenediamine, oxalyl diamine, and 1,4-phenylenediamine.

[0029] In some embodiments of the present invention, the method for preparing a RAFT agent comprises: making a nitrogen heterocyclic organic molecule negatively charged under alkaline conditions, mixing the negatively charged nitrogen heterocyclic organic molecule with carbon disulfide, forming a thioester molecule with a negatively charged group by attacking the carbon-sulfur double bond in the carbon disulfide, mixing the thioester molecule with a benzyl halide-containing molecule, and allowing the negatively charged group of the thioester molecule to attack the benzyl halide site of the benzyl halide-containing molecule to form a RAFT agent;

[0030] Preferably, the nitrogen heterocyclic organic molecule includes any one of pyrrole, carbazole, and indole;

[0031] Preferably, the benzyl halide-containing molecule includes any one of benzyl chloride, benzyl bromide and benzyl iodide.

[0032] The third aspect of the present invention provides a method for improving the drug loading performance of drug-loaded microspheres, wherein the drug-loaded microspheres are added to a mixed solution of the polymer that can be assembled with proteins and hydrophobic small molecule drugs and the drug to be loaded.

[0033] The diblock polymer prepared by the method of the present invention that can be assembled with hydrophobic small molecules and protein drugs is in a dissolved state for hydrophobic small molecule drugs such as sorafenib and lenvatinib in a DMSO solution. When water is added to the system, the insoluble drug is quickly assembled with the cavity-containing molecules in the diblock polymer, and after assembly, the assembly block is made hydrophobic, and finally forms a vesicle structure in an aqueous solution under the action of the cationic block. The outer layer of the vesicle structure has a large number of hydrophilic amino cations, which interact with the negatively charged sulfonic acid groups on the microsphere polymer skeleton, so that the microspheres can load hydrophobic small molecule drugs such as sorafenib and lenvatinib.

[0034] For water-soluble protein drugs such as atelizumab, the diblock polymer is attached to the protein surface through multiple cavity structures in the cyclodextrin block in the aqueous solution, thereby forming a layer of diblock polymer on the surface of the protein molecule. The cationic groups in the polymer interact with the negatively charged sulfonic acid groups on the microsphere polymer skeleton, so that the microspheres can load protein drugs such as atelizumab.

[0035] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present disclosure. DETAILED DESCRIPTION

[0036] The following will be combined with the embodiments to clearly and completely describe the concept of the present invention and the technical effects produced, so as to fully understand the purpose, characteristics and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present invention.

[0037] The experimental methods without specific conditions in the examples are usually carried out according to conventional conditions in the art or conditions recommended by the manufacturers; the raw materials and reagents used, unless otherwise specified, are all raw materials and reagents that can be obtained from commercial channels such as conventional markets.

[0038] In the embodiment, the main molecular polymer containing the cavity structure is polymerized by a monomer containing a cavity functional group. In a specific embodiment, the preparation of poly(propyl methacrylate)-2-hydroxy-3-methylamino-α-cyclodextrin comprises the following steps:

[0039] Step 1: Take 15g (13.2mmol) of α-cyclodextrin monomer with cavity functional group and put it into 125mL of purified water, slowly add NaOH solution (5mL, 41mmol) and stir vigorously until α-cyclodextrin is completely dissolved. Dissolve 2.5g (13.2mmol) of p-toluenesulfonyl chloride in 7.5mL of acetonitrile solution and slowly add it dropwise into the reaction system. o C for 3 h. After the reaction was completed, the solution was filtered under reduced pressure to remove solids and the filtrate was collected. The filtrate was distilled under reduced pressure and acetone was added for sedimentation. After the sedimentation was completed, the solid was collected by filtration under reduced pressure. The solid was placed at 40 o C in a vacuum drying oven overnight.

[0040] Step 2: Mix 3 g of the product from step 1 and 62.5 mL of 1,3-propylenediamine at 80 °C. o The reaction was stirred at 40 °C for 6 h. The reaction solution was distilled under reduced pressure and then acetone was added for precipitation. After the precipitation was completed, the solid was collected by vacuum filtration. The solid was placed at 40 o C in a vacuum drying oven overnight.

[0041] Step 3: Dissolve 0.88 g of the product from step 2 and 1.64 mL of glycidyl methacrylate in 10 mL of N,N-dimethylformamide, introduce N2 for 30 min, and heat the system to 60 o After the reaction was completed, the reaction solution was added dropwise to excess acetone for precipitation. After the precipitation was completed, the solid was collected by vacuum filtration. The solid was placed at 40 o C in a vacuum drying oven overnight to obtain propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin.

[0042] The difference between the preparation method of poly(propyl methacrylate)-2-hydroxy-3-methylamino-β-cyclodextrin and poly(propyl methacrylate)-2-hydroxy-3-methylamino-γ-cyclodextrin and the preparation method of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin is that β-cyclodextrin or γ-cyclodextrin is used instead of α-cyclodextrin in the same molar amount.

[0043] The molar ratio of any one of the ammonium functional group-containing monomer, the amine functional group-containing monomer, and the epoxy functional group-containing monomer to the cavity functional group-containing monomer or the cavity-containing organic molecule is (0.25-4):1.

[0044] Example 1: Preparation of polymer A1 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0045]

[0046] Step 1: Take 40 μL of benzyl chloride, 0.34 mg of copper bromide, 6.5 μL of pentamethyldiethyltriamine, 400 mg of N-(3-aminopropyl)methacrylamide hydrochloride and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg of cuprous bromide and continue to pass high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain a cationic polymer.

[0047] Step 2: Take 0.34 mg of copper bromide, 6.5 μL of pentamethyldiethyltriamine, 4.5 mg of cationic polymer, 56 mg of 2-hydroxy-3-methylamino-α-cyclodextrin propyl methacrylate and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg of cuprous bromide and continue to pass high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0048] Example 2-Example 9: Preparation of polymers A2-A9 that can be assembled with proteins and hydrophobic small molecule drugs by ATRP reaction

[0049] The difference between the preparation method of Example 2-Example 9 and that of Example 1 is that the same molar amounts of 2-aminoethyl methacrylate hydrochloride, allylamine hydrochloride, 2-methylallylamine hydrochloride, O-allylhydroxylamine hydrochloride, 3-butenylamine hydrochloride, 3-methylbut-2-en-1-amine hydrochloride, 2-allylaniline hydrochloride, and N-(3-aminopropyl)methacrylamide hydrochloride are used in sequence instead of N-(3-aminopropyl)methacrylamide hydrochloride;

[0050] Example 10: Preparation of polymer A10 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0051] The difference between the preparation method and Example 1 is that the same molar amount of polypropyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, and the molar amount of the monomer containing the ammonium functional group to the molar amount of the monomer with the cavity functional group is 0.25:1, that is, 2.3 mg of cationic polymer and 64 mg of propyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin are used instead of 4.5 mg of cationic polymer and 56 mg of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin in Example 1.

[0052] Example 11: Preparation of polymer A11 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0053] The preparation method is different from that in Example 1 only in that the same molar amount of polypropyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, and the ratio of the molar amount of the monomer containing the ammonium functional group to the molar amount of the monomer with the cavity functional group is 1:0.25, that is, 9 mg of cationic polymer and 18 mg of propyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin are used instead of 4.5 mg of cationic polymer and 56 mg of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin in Example 1.

[0054] Example 12: Preparation of polymer A12 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0055]

[0056] Step 1: Take 40 μL of benzyl chloride, 0.34 mg of copper bromide, 6.5 μL of pentamethyldiethyltriamine, 400 mg of N-(3-aminopropyl)methacrylamide hydrochloride and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg of cuprous bromide and continue to pass high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain a cationic polymer.

[0057] Step 2: Take 4.5 mg of the above product, 5 mg of hydroxyethyl methacrylate, 2.5 mg of azobisisobutyl, and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 1 hour. Heat to 65 oC for 8 hours. The resulting solution was added with n-hexane to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven and dried for 12 hours to obtain a diblock polymer.

[0058] Step 3: Dissolve 60 mg of 4-azosulfonic acid sodium calix[4]arene in 10 mL of anhydrous DMF, and add 1 g of sodium carbonate (100 mg / mL) and 30 g of epibromopropane in sequence. o After stirring at 25 °C for 24 h, the modified 4-azosulfonic acid sodium calix[4]arene was obtained by ether precipitation. 8 mg of the product of step 2 was dissolved in 5 mL of sodium carbonate buffer (100 mM, pH = 8.5), 30 mg of the modified 4-azosulfonic acid sodium calix[4]arene was added, and the mixture was stirred at 25 °C for 24 h. o The mixture was stirred at C for 24 h. Unreacted calixarene was removed by dialysis against deionized water, and polymers that can be assembled with proteins and hydrophobic small molecule drugs were obtained by freeze drying.

[0059] Example 13: Preparation of polymer A13 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0060] The difference between the preparation method of Example 13 and Example 12 is that the same molar amount of 4-azosulfonato sodium calix[6]arene is used to replace the 4-azosulfonato sodium calix[4]arene in Example 12.

[0061] Example 14: Preparation of polymer A14 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0062]

[0063] Step 1: Take 40 μL benzyl chloride, 0.34 mg copper bromide, 6.5 μL pentamethyldiethyltriamine, 160 mg 2-methylallylamine and 1 mL N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg cuprous bromide and continue passing high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain an amine-containing polymer.

[0064] Step 2: Take 0.34 mg of copper bromide, 6.5 μL of pentamethyldiethyltriamine, 2 mg of amine-containing polymer, 56 mg of 2-hydroxy-3-methylamino-α-cyclodextrin propyl methacrylate and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg of cuprous bromide and continue to pass high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven and dried for 12 hours to obtain a diblock polymer.

[0065] Step 3: Weigh 0.5 g of diblock polymer and dissolve it in 10 mL of dimethyl sulfoxide (or a corresponding solvent that can dissolve the compound), stir and dissolve it with a magnetic stirrer (500 rpm, 30 min) to completely dissolve it. Under ice bath conditions, slowly add hydrochloric acid (38%) or acetic acid, continuously monitor the pH of the reaction environment, and control the pH value at 1-3. After reaching the predetermined pH range, continue to stir the reaction for 1 h. After the reaction is completed, use anhydrous ether to precipitate and centrifuge the polymer. After repeating 2 to 5 times, vacuum dry the obtained solid sample to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0066] Example 15-Example 23: Preparation of polymers A14-A22 that can be assembled with proteins and hydrophobic small molecule drugs by ATRP reaction

[0067] The difference between the preparation methods of Examples 15 to 23 and Example 14 is that the same molar amounts of acrylamide, N-(2-amino-2-oxoethyl)acrylamide, 2-buteneamide, 4-aminostilbene, 3-aminostyrene, 4-aminostyrene, methacrylamide, 2-methylallylamine, and 4-vinylbenzylamine are used in sequence instead of 2-methylallylamine;

[0068] Example 24: Preparation of polymer A24 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0069] The preparation method is different from that of Example 14 only in that the same molar amount of polypropyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, and the ratio of the molar amount of the monomer containing the amino functional group to the molar amount of the monomer with the cavity functional group is 0.25:1, that is, 1 mg of the amino-containing polymer and 64 mg of propyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin are used instead of 2 mg of the amino-containing polymer and 56 mg of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin in Example 14.

[0070] Example 25: Preparation of polymer A25 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0071] The preparation method is different from that of Example 14 only in that the same molar amount of polypropyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, and the ratio of the molar amount of the monomer containing the ammonium functional group to the molar amount of the monomer with the cavity functional group is 1:0.25, that is, 4 mg of the amino-containing polymer and 18 mg of propyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin are used instead of 2 mg of the amino-containing polymer and 56 mg of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin in Example 14.

[0072] Example 26: Preparation of polymer A26 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0073]

[0074] Step 1: Take 40 μL benzyl chloride, 0.34 mg copper bromide, 6.5 μL pentamethyldiethyltriamine, 160 mg 2-methylallylamine and 1 mL N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg cuprous bromide and continue passing high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain an amine-containing polymer.

[0075] Step 2: Take 2 mg of the above solid product, 5 mg of allyl 4-hydroxybenzoate, 2.5 mg of azobisisobutyl, and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle, and pass high-purity nitrogen for 1 hour. Heat to 65 oC for 8 hours. The resulting solution was added with n-hexane to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain a diblock polymer precursor.

[0076] Step 3: Dissolve 60 mg of 4-azosulfonic acid sodium calix[4]arene in 10 mL of anhydrous DMF, and add 1 g of sodium carbonate (100 mg / mL) and 30 g of epibromopropane in sequence. o After stirring at 25 °C for 24 h, poly (4-azosulfonato sodium calix[4]arene) was obtained by ether precipitation. 8 mg of the product of step 2 was dissolved in 5 mL of sodium carbonate buffer (100 mM, pH = 8.5), 30 mg of epibromohydrin-modified poly (4-azosulfonato sodium calix[4]arene) was added, and the mixture was stirred at 25 °C for 24 h. o C for 24 h. The unreacted calixarene was removed by dialysis against deionized water, and the diblock polymer was obtained by freeze-drying.

[0077] Step 4: Weigh 0.5 g of diblock polymer and dissolve it in 10 mL of dimethyl sulfoxide (or a corresponding solvent that can dissolve the compound), stir and dissolve it with a magnetic stirrer (500 rpm, 30 min) to completely dissolve it. Under ice bath conditions, slowly add hydrochloric acid (38%) or acetic acid, continuously monitor the pH of the reaction environment, and control the pH value at 1-3. After reaching the predetermined pH range, continue to stir the reaction for 1 h. After the reaction is completed, use anhydrous ether to precipitate and centrifuge the polymer. After repeating 2 to 5 times, vacuum dry the obtained solid sample to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0078] Example 27: Preparation of polymer A27 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0079] The only difference between the preparation method of Example 27 and Example 26 is that the same molar amount of 4-azosulfonato sodium calix[6]arene is used to replace the 4-azosulfonato sodium calix[4]arene in Example 12.

[0080] Example 28: Preparation of polymer A28 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0081]

[0082] Step 1: Take 40 μL of benzyl chloride, 0.34 mg of copper bromide, 6.5 μL of pentamethyldiethyltriamine, 320 mg of glycidyl methacrylate and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg of cuprous bromide and continue passing high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain an epoxy group-containing polymer.

[0083] Step 2: Take 0.34 mg of copper bromide, 6.5 μL of pentamethyldiethyltriamine, 3.5 mg of epoxy-containing polymer, 56 mg of 2-hydroxy-3-methylamino-α-cyclodextrin propyl methacrylate and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg of cuprous bromide and continue to pass high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven and dried for 12 hours to obtain a diblock polymer.

[0084] Step 3: Take 0.5 mL of propylene diamine and 5 mL of N,N-dimethylformamide or N,N-dimethylacetamide and add them to a 10 mL reaction bottle and stir for 10 minutes. Then add 50 mg of the above product and o The reaction was stirred at 40 °C for 7 hours. The resulting solution was precipitated with ether, and the precipitated solution was centrifuged to obtain a solid product. The solid product was purified by dialysis in purified water. The purified solid product was dried by freeze drying.

[0085] Step 4: Weigh 0.5 g of diblock polymer and dissolve it in 10 mL of dimethyl sulfoxide (or a corresponding solvent that can dissolve the compound), stir and dissolve it with a magnetic stirrer (500 rpm, 30 min) to completely dissolve it. Under ice bath conditions, slowly add hydrochloric acid (38%) or acetic acid, continuously monitor the pH of the reaction environment, and control the pH value at 1-3. After reaching the predetermined pH range, continue to stir the reaction for 1 h. After the reaction is completed, use anhydrous ether to precipitate and centrifuge the polymer. After repeating 2 to 5 times, vacuum dry the obtained solid sample to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0086] Example 29-Example 37: Preparation of polymers A29-A37 that can be assembled with proteins and hydrophobic small molecule drugs by ATRP reaction

[0087] The difference between the preparation methods of Examples 29 to 37 and Example 28 is that the same molar amounts of allyl glycidyl ether, 3,4-epoxy-1-butene, (3,4-epoxycyclohexyl) methyl acrylate, 1,2-epoxy-9-decene, 1,2-epoxy-5-hexene, 1,2-epoxy-4-vinylcyclohexane, glycidyl acrylate, glycidyl methacrylate, and 3,4-epoxy-1-butene allyl glycidyl ether are used in sequence instead of glycidyl methacrylate;

[0088] Example 38-Example 85: Preparation of polymers A38-A85 that can be assembled with proteins and hydrophobic small molecule drugs by ATRP reaction

[0089] The difference between the preparation methods of Examples 38 to 85 and those of Example 28 is that the same molar amounts of 1,3-propylenediamine, p-phenylenediamine, adipic acid diamine, N,N'-bis(2-aminoethyl)-1,3-propylenediamine, 1,3-bis(aminomethyl)cyclohexane, N,N'-bis(3-aminopropyl)ethylenediamine, 6,6'-iminodihexylamine, 1,4-butanediol bis(3-aminopropyl) ether, 2-chloro-5-methyl-1,4-phenylenediamine, 2-chloro-5-nitro 1,4-phenylenediamine, 2-chloro-1,4-phenylenediamine, 1,3-cyclohexanediamine, 1,4-cyclohexanediamine, 2,6-diaminoanthraquinone, 6,6'-diamino-2,2'-bipyridine, 1,4-butanediamine, 3,6-diaminocarbazole, 1,10-diaminodecane, 3,3'-diaminodipropylamine, 1,12-diaminododecane, 2,7-diaminofluorene, 1,7-heptanediamine, 1,6-hexanediamine, 2,2'-diamino-N-methyldiethylamine Amine, 3,3'-diamino-N-methyldipropylamine, 1,2-diamino-2-methylpropane, 1,8-diaminooctane, 1,3-diaminopentane, 1,5-diaminopentane, 1,2-diaminopropane, 1,3-propylenediamine, 2-hydroxy-1.3-diaminopropane, 1,14-diamino-3,6,9,12-tetraoxatetradecane, 2,5-diaminotoluene, 1,11-diamino-3,6,9-trioxaundecane, 1,11-diaminodecane oxadiazole, 4,4'-diamino-3,3'-dimethoxybiphenyl, 2,6-dibromo-1,4-phenylenediamine, 2,5-dichloro-1,4-phenylenediamine, 2,2'-diaminodiethylamine, 3,3'-dihydroxybenzidine, 2,5-dimethyl-1,4-phenylenediamine, 2-methyl-1,5-diaminopentane, 2-methyl-1,3-propylenediamine, naphthalene-2,6-diamine, 2-nitro-1,4-phenylenediamine, oxalyl diamine, 1,4-phenylenediamine instead of diethylamine;

[0090] Example 86: Preparation of polymer A86 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0091] The preparation method differs from that of Example 28 only in that the same molar amount of polypropyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, and the ratio of the molar amount of the monomer containing the alkylene oxide functional group to the molar amount of the monomer with the cavity functional group is 0.25:1, that is, 1.8 mg of epoxy-containing polymer and 64 mg of propyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin are used instead of 3.5 mg of epoxy-containing polymer and 56 mg of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin in Example 28.

[0092] Example 87: Preparation of polymer A87 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0093] The difference between the preparation method and Example 28 is that the same molar amount of polypropyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, and the ratio of the molar amount of the monomer containing the alkylene oxide functional group to the molar amount of the monomer with the cavity functional group is 1:0.25, that is, 7.2 mg of epoxy-containing polymer and 18 mg of propyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin are used instead of 3.5 mg of epoxy-containing polymer and 56 mg of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin in Example 28.

[0094] Example 88: Preparation of polymer A88 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0095]

[0096] Step 1: Take 40 μL of benzyl chloride, 0.34 mg of copper bromide, 6.5 μL of pentamethyldiethyltriamine, 320 mg of glycidyl methacrylate and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. Pass high-purity nitrogen for 30 minutes. Add 4.28 mg of cuprous bromide and continue passing high-purity nitrogen for 30 minutes. Heat to 65 o C, stirred and reacted for 14 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain an epoxy group-containing polymer.

[0097] Step 2: Take 3.5 mg of the above solid product, 5 mg of 6-(allyloxycarbonylamino)-1-hexanol, 2.5 mg of azobisisobutyl, and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle, and pass high-purity nitrogen for 1 hour. Heat to 65 o C for 8 hours. The resulting solution was added with n-hexane to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain a diblock polymer precursor.

[0098] Step 3: Dissolve 60 mg of 4-azosulfonic acid sodium calix[4]arene in 10 mL of anhydrous DMF, and add 1 g of sodium carbonate (100 mg / mL) and 30 g of epibromopropane in sequence. o After stirring at 25 °C for 24 h, the modified sodium 4-azosulfonate calix[4]arene was obtained by ether precipitation. 8 mg of the product of step 2 was dissolved in 5 mL of sodium carbonate buffer (100 mM, pH = 8.5), 30 mg of epibromohydrin-modified 4-azosulfonate calix[4]arene was added, and the mixture was stirred at 25 °C for 24 h. o C for 24 h. The unreacted calixarene was removed by dialysis against deionized water, and the diblock polymer was obtained by freeze-drying.

[0099] Step 4: Take 0.5 mL of propylene diamine and 5 mL of N,N-dimethylformamide or N,N-dimethylacetamide and add them to a 10 mL reaction bottle and stir for 10 minutes. Then add 50 mg of the above diblock polymer and stir at 60 °C. o The reaction was stirred at 40 °C for 7 hours. The resulting solution was precipitated with ether, and the precipitated solution was centrifuged to obtain a solid product. The solid product was purified by dialysis in purified water. The purified solid product was dried by freeze drying.

[0100] Step 5: Weigh 0.5 g of diblock polymer and dissolve it in 10 mL of dimethyl sulfoxide (or a corresponding solvent that can dissolve the compound), stir and dissolve it with a magnetic stirrer (500 rpm, 30 min) to completely dissolve it. Under ice bath conditions, slowly add hydrochloric acid (38%) or acetic acid, continuously monitor the pH of the reaction environment, and control the pH value at 1-3. After reaching the predetermined pH range, continue to stir the reaction for 1 h. After the reaction is completed, use anhydrous ether to precipitate and centrifuge the polymer. After repeating 2 to 5 times, vacuum dry the obtained solid sample to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0101] Example 89: Preparation of polymer A89 capable of assembling with proteins and hydrophobic small molecule drugs by ATRP reaction

[0102] The difference between the preparation method of Example 89 and that of Example 88 is that the same molar amount of 4-sodium azosulfonate calix[6]arene is used to replace the 4-sodium azosulfonate calix[4]arene in Example 12.

[0103] Example 90: Preparation of RAFT agent

[0104] Take 117.8 mg of potassium hydroxide and add anhydrous dimethyl sulfoxide and stir vigorously until the potassium hydroxide dissolves. Continue to add 145 μL of pyrrole and stir for 2 hours. Slowly drop 150 μL of carbon disulfide and continue stirring for 5 hours. Add 242 μL of benzyl chloride and stir at room temperature for 36 hours. Add excess acetone to the resulting red solution to obtain a suspension. Filter the solid and precipitate at 40 o C vacuum drying oven to obtain the final product.

[0105] Preferably, the nitrogen heterocyclic organic molecule includes any one of pyrrole, carbazole, and indole;

[0106] Preferably, the benzyl halide-containing molecule includes any one of benzyl chloride, benzyl bromide and benzyl iodide.

[0107] Example 91: Preparation of polymer A91 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0108]

[0109] Step 1: Take 3.5 mg of 2-phenyl-2-propylbenzodisulfide, 0.7 g of N-(3-aminopropyl)methacrylamide hydrochloride, 2.5 mg of azobisisobutyronitrile and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. o C for 8 hours. After the obtained solution is cooled, tetrahydrofuran is added for precipitation, and N,N-dimethylformamide / tetrahydrofuran is used to repeat the dissolution / precipitation for 2 times. The obtained solid is separated from the solution by centrifugation and dried in a vacuum drying oven.

[0110] Step 2: Take 4.5 mg of the above solid product, 56 mg of 2-hydroxy-3-methylamino-α-cyclodextrin propyl methacrylate, 2.5 mg of azobisisobutyl, and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle, and pass high-purity nitrogen for 1 hour. Heat to 65 o C for 8 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0111] Example 92-Example 99: Preparation of polymers A92-A99 that can be assembled with proteins and hydrophobic small molecule drugs by RAFT reaction

[0112] The difference between the preparation method of Example 92-Example 99 and that of Example 88 is that: the same weight of 2-aminoethyl methacrylate hydrochloride, allylamine hydrochloride, 2-methylallylamine hydrochloride, O-allylhydroxylamine hydrochloride, 3-butenylamine hydrochloride, 3-methylbut-2-en-1-amine hydrochloride, 2-allylaniline hydrochloride, and N-(3-aminopropyl)methacrylamide hydrochloride are used in sequence instead of N-(3-aminopropyl)methacrylamide hydrochloride;

[0113] Example 100: Preparation of polymer A100 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0114] The preparation method differs from that of Example 91 only in that the same molar amount of poly(propyl methacrylate)-2-hydroxy-3-methylamino-β-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin;

[0115] Example 101: Preparation of polymer A101 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0116] The preparation method differs from that of Example 91 only in that the same molar amount of poly(propyl methacrylate)-2-hydroxy-3-methylamino-γ-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin.

[0117] Example 102: Preparation of polymer A102 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0118]

[0119] Step 1: Take 3.5 mg of 2-phenyl-2-propylbenzodisulfide, 0.7 g of N-(3-aminopropyl)methacrylamide hydrochloride, 2.5 mg of azobisisobutyronitrile and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle. o C for 8 hours. After the obtained solution is cooled, tetrahydrofuran is added for precipitation, and N,N-dimethylformamide / tetrahydrofuran is used to repeat the dissolution / precipitation for 2 times. The obtained solid is separated from the solution by centrifugation and dried in a vacuum drying oven.

[0120] Step 2: Take 4.5 mg of the above solid product, 5 mg of 3-hexenol lactate, 2.5 mg of azobisisobutyl, and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle, and pass high-purity nitrogen for 1 hour. Heat to 65 o C for 8 hours. The resulting solution was added with n-hexane to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven and dried for 12 hours to obtain a diblock polymer.

[0121] Step 3: Dissolve 60 mg of 4-azosulfonic acid sodium calix[4]arene in 10 mL of anhydrous DMF, and add 1 g of sodium carbonate (100 mg / mL) and 30 g of epibromopropane in sequence. o After stirring at 25 °C for 24 h, the modified sodium 4-azosulfonate calix[4]arene was obtained by ether precipitation. 8 mg of the product of step 2 was dissolved in 5 mL of sodium carbonate buffer (100 mM, pH = 8.5), 30 mg of epibromohydrin-modified 4-azosulfonate calix[4]arene was added, and the mixture was stirred at 25 °C for 24 h. o The mixture was stirred at C for 24 h. Unreacted calixarene was removed by dialysis against deionized water, and the product was obtained by lyophilization.

[0122] Example 103: Preparation of polymer A103 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0123] The difference between the preparation method of Example 103 and that of Example 102 is that the same molar amount of 4-sodium azosulfonate calix[6]arene is used to replace the 4-sodium azosulfonate calix[4]arene in Example 12.

[0124] Example 104: Preparation of polymer A104 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0125]

[0126] Step 1: Add 3.5 mg of 2-phenyl-2-propylbenzodisulfide, 160 mg of 2-methylallylamine, 2.5 mg of azobisisobutyronitrile and 1 mL of N,N-dimethylformamide into a 10 mL reaction bottle. o C for 8 hours. After the obtained solution is cooled, tetrahydrofuran is added for precipitation, and N,N-dimethylformamide / tetrahydrofuran is used to repeat the dissolution / precipitation for 2 times. The obtained solid is separated from the solution by centrifugation and dried in a vacuum drying oven to obtain an amine-containing polymer.

[0127] Step 2: Take 2 mg of the above solid product, 56 mg of 2-hydroxy-3-methylamino-α-cyclodextrin propyl methacrylate, 2.5 mg of azobisisobutyl, and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle, and pass high-purity nitrogen for 1 hour. Heat to 65 o C for 8 hours. Tetrahydrofuran was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven and dried for 12 hours to obtain a diblock polymer.

[0128] Step 3: Weigh 0.5 g of diblock polymer and dissolve it in 10 mL of dimethyl sulfoxide (or a corresponding solvent that can dissolve the compound), stir and dissolve it with a magnetic stirrer (500 rpm, 30 min) to completely dissolve it. Under ice bath conditions, slowly add hydrochloric acid (38%) or acetic acid, continuously monitor the pH of the reaction environment, and control the pH value at 1-3. After reaching the predetermined pH range, continue to stir the reaction for 1 h. After the reaction is completed, use anhydrous ether to precipitate and centrifuge the polymer. After repeating 2 to 5 times, vacuum dry the obtained solid sample to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0129] Example 105-Example 113: Preparation of polymers A101-A109 capable of being assembled with proteins and hydrophobic small molecule drugs by RAFT reaction

[0130] The difference between the preparation methods of Examples 105 to 113 and Example 104 is that the same molar amounts of acrylamide, N-(2-amino-2-oxoethyl)acrylamide, 2-buteneamide, 4-aminostilbene, 3-aminostyrene, 4-aminostyrene, methacrylamide, 2-methylallylamine, and 4-vinylbenzylamine are used in sequence to replace 2-methylallylamine;

[0131] Example 114: Preparation of polymer A114 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0132] The preparation method is different from Example 104 only in that the same molar amount of poly(propyl methacrylate)-2-hydroxy-3-methylamino-β-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin.

[0133] Example 115: Preparation of polymer A115 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0134] The preparation method is different from Example 104 only in that the same molar amount of poly(propyl methacrylate)-2-hydroxy-3-methylamino-γ-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin.

[0135] Example 116: Preparation of polymer A116 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0136]

[0137] Step 1: Add 3.5 mg of 2-phenyl-2-propylbenzodisulfide, 160 mg of 2-methylallylamine, 2.5 mg of azobisisobutyronitrile and 1 mL of N,N-dimethylformamide into a 10 mL reaction bottle. o C for 8 hours. After the obtained solution is cooled, tetrahydrofuran is added for precipitation, and N,N-dimethylformamide / tetrahydrofuran is used to repeat the dissolution / precipitation for 2 times. The obtained solid is separated from the solution by centrifugation and dried in a vacuum drying oven to obtain an amine-containing polymer.

[0138] Step 2: Take 2 mg of the above solid product, 5 mg of hydroxyethyl acrylate, 2.5 mg of azobisisobutyl, and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle, and introduce high-purity nitrogen for 1 hour. Heat to 65 o C for 8 hours. The resulting solution was added with n-hexane to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven for 12 hours to obtain a diblock polymer precursor.

[0139] Step 3: Dissolve 60 mg of 4-azosulfonic acid sodium calix[4]arene in 10 mL of anhydrous DMF, and add 1 g of sodium carbonate (100 mg / mL) and 30 g of epibromopropane in sequence. o After stirring at 25 °C for 24 h, the modified sodium 4-azosulfonate calix[4]arene was obtained by ether precipitation. 8 mg of the product of step 2 was dissolved in 5 mL of sodium carbonate buffer (100 mM, pH = 8.5), 30 mg of epibromohydrin-modified 4-azosulfonate calix[4]arene was added, and the mixture was stirred at 25 °C for 24 h. o C for 24 h. The unreacted calixarene was removed by dialysis against deionized water, and the diblock polymer was obtained by freeze-drying.

[0140] Step 4: Weigh 0.5 g of diblock polymer and dissolve it in 10 mL of dimethyl sulfoxide (or a corresponding solvent that can dissolve the compound), stir and dissolve it with a magnetic stirrer (500 rpm, 30 min) to completely dissolve it. Under ice bath conditions, slowly add hydrochloric acid (38%) or acetic acid, continuously monitor the pH of the reaction environment, and control the pH value at 1-3. After reaching the predetermined pH range, continue to stir the reaction for 1 h. After the reaction is completed, use anhydrous ether to precipitate and centrifuge the polymer. After repeating 2 to 5 times, vacuum dry the obtained solid sample to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0141] Example 117: Preparation of polymer A117 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0142] The difference between the preparation method of Example 117 and Example 116 is that the same molar amount of 4-azosulfonato sodium calix[6]arene is used to replace the 4-azosulfonato sodium calix[4]arene in Example 12.

[0143] Example 118: Preparation of polymer A118 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0144]

[0145] Step 1: Add 3.5 mg of 2-phenyl-2-propylbenzodisulfide, 320 mg of glycidyl methacrylate, 2.5 mg of azobisisobutyronitrile and 1 mL of N,N-dimethylformamide into a 10 mL reaction bottle. o C for 8 hours. After the obtained solution is cooled, acetone is added for precipitation, and dimethyl sulfoxide / acetone is used to repeat the dissolution / precipitation for 2 times. The obtained solid is separated from the solution by centrifugation and dried in a vacuum drying oven to obtain an epoxy-containing polymer.

[0146] Step 2: Take 3.5 mg of the above solid product, 56 mg of 2-hydroxy-3-methylamino-α-cyclodextrin propyl methacrylate, and 2.5 mg of diisobutyl azodicarbonyl, add them into a 10 mL reaction bottle, and pass high-purity nitrogen for 1 hour. Heat to 65 o C for 8 hours. Acetone was added to the obtained solution to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed in a vacuum oven and dried for 12 hours to obtain a diblock polymer.

[0147] Step 3: Take 0.1 mL of diethylamine and 5 mL of N,N-dimethylformamide or N,N-dimethylacetamide and add them to a 10 mL reaction bottle. Stir for 10 minutes. Then add 50 mg of the above product at 60o The reaction was stirred at 40 °C for 6 hours. The obtained solution was precipitated with acetone, and the precipitated solution was separated by centrifugation to obtain a solid product. The solid product was dried by freeze drying to obtain an aminated diblock polymer;

[0148] Step 4: Weigh 0.5 g of the aminated diblock polymer and dissolve it in 10 mL of dimethyl sulfoxide (or a corresponding solvent that can dissolve the compound), stir and dissolve it with a magnetic stirrer (500 rpm, 30 min) to completely dissolve it. Under ice bath conditions, slowly add hydrochloric acid (38%) or acetic acid, continuously monitor the pH of the reaction environment, and control the pH value at 1-3. After reaching the predetermined pH range, continue to stir the reaction for 1 h. After the reaction is completed, use anhydrous ether to precipitate and centrifuge the polymer. After repeating 2 to 5 times, vacuum dry the obtained solid sample to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0149] Examples 119 to 127: Preparation of polymers A119 to A127 capable of being assembled with proteins and hydrophobic small molecule drugs by RAFT reaction

[0150] The difference between the preparation methods of Examples A119 to 127 and those of Example 118 is that the same molar amounts of allyl glycidyl ether, 3,4-epoxy-1-butene, (3,4-epoxycyclohexyl) methyl acrylate, 1,2-epoxy-9-decene, 1,2-epoxy-5-hexene, 1,2-epoxy-4-vinylcyclohexane, glycidyl acrylate, glycidyl methacrylate, and 3,4-epoxy-1-butene allyl glycidyl ether are used in sequence instead of glycidyl methacrylate;

[0151] Example 128-Example 175: Preparation of polymers A128-A175 that can be assembled with proteins and hydrophobic small molecule drugs by RAFT reaction

[0152] The difference between the preparation methods of Examples A128 to 175 and those of Example 118 is that the same molar amounts of 1,3-propylenediamine, p-phenylenediamine, adipic acid diamine, N,N'-bis(2-aminoethyl)-1,3-propylenediamine, 1,3-bis(aminomethyl)cyclohexane, N,N'-bis(3-aminopropyl)ethylenediamine, 6,6'-iminodihexylamine, 1,4-butanediol bis(3-aminopropyl) ether, 2-chloro-5-methyl-1,4-phenylenediamine, 2-chloro- 5-nitro-1,4-phenylenediamine, 2-chloro-1,4-phenylenediamine, 1,3-cyclohexanediamine, 1,4-cyclohexanediamine, 2,6-diaminoanthraquinone, 6,6'-diamino-2,2'-bipyridine, 1,4-butanediamine, 3,6-diaminocarbazole, 1,10-diaminodecane, 3,3'-diaminodipropylamine, 1,12-diaminododecane, 2,7-diaminofluorene, 1,7-heptanediamine, 1,6-hexanediamine, 2,2'-diamino-N-methyl Diethylamine, 3,3'-diamino-N-methyldipropylamine, 1,2-diamino-2-methylpropane, 1,8-diaminooctane, 1,3-diaminopentane, 1,5-diaminopentane, 1,2-diaminopropane, 1,3-propylenediamine, 2-hydroxy-1.3-diaminopropane, 1,14-diamino-3,6,9,12-tetraoxatetradecane, 2,5-diaminotoluene, 1,11-diamino-3,6,9-trioxaundecane, 1,11-diamino Undecane, 4,4'-diamino-3,3'-dimethoxybiphenyl, 2,6-dibromo-1,4-phenylenediamine, 2,5-dichloro-1,4-phenylenediamine, 2,2'-diaminodiethylamine, 3,3'-dihydroxybenzidine, 2,5-dimethyl-1,4-phenylenediamine, 2-methyl-1,5-diaminopentane, 2-methyl-1,3-propylenediamine, naphthalene-2,6-diamine, 2-nitro-1,4-phenylenediamine, oxalyl diamine, 1,4-phenylenediamine instead of diethylamine;

[0153] Example 176: Preparation of polymer A176 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0154] The preparation method differs from that of Example 118 only in that the same molar amount of poly(propyl methacrylate)-2-hydroxy-3-methylamino-β-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin;

[0155] Example 177: Preparation of polymer A177 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0156] The preparation method is different from that of Example 113 only in that the same molar amount of poly(propyl methacrylate)-2-hydroxy-3-methylamino-γ-cyclodextrin is used instead of propyl methacrylate-2-hydroxy-3-methylamino- α-Cyclodextrins.

[0157] Example 178: Preparation of polymer A178 that can be assembled with proteins and hydrophobic small molecule drugs by RAFT reaction

[0158]

[0159] Step 1: Take 3.5 mg of 2-phenyl-2-propylbenzodisulfide, 320 mg of glycidyl methacrylate, and 2.5 mg of azobisisobutyronitrile and add them to a 10 mL reaction bottle. o C for 8 hours. After the obtained solution is cooled, tetrahydrofuran is added for precipitation, and N,N-dimethylformamide / tetrahydrofuran is used to repeat the dissolution / precipitation for 2 times. The obtained solid is separated from the solution by centrifugation and dried in a vacuum drying oven to obtain an epoxy-containing polymer.

[0160] Step 2: Take 3.5 mg of the above solid product, 5 mg of 6-hydroxyhexyl methacrylate or hydroxypropyl acrylate, 2.5 mg of azobisisobutyl, and 1 mL of N,N-dimethylformamide and add them to a 10 mL reaction bottle, and pass high-purity nitrogen for 1 hour. Heat to 65 o C for 8 hours. The resulting solution was added with n-hexane to obtain a suspension, and the suspension was centrifuged at 5000 rpm to obtain a solid sample. The above steps were repeated 3 times, and the solid was placed at 40 o C vacuum oven and dried for 12 hours to obtain a diblock polymer.

[0161] Step 3: Dissolve 60 mg of 4-azosulfonic acid sodium calix[4]arene in 10 mL of anhydrous DMF, and add 1 g of sodium carbonate (100 mg / mL) and 30 g of epibromopropane in sequence. o After stirring at 25 °C for 24 h, the modified sodium 4-azosulfonate calix[4]arene was obtained by ether precipitation. 8 mg of the product of step 2 was dissolved in 5 mL of sodium carbonate buffer (100 mM, pH = 8.5), 30 mg of epibromohydrin-modified 4-azosulfonate calix[4]arene was added, and the mixture was stirred at 25 °C for 24 h. o The mixture was stirred at C for 24 h. Unreacted calixarene was removed by dialysis against deionized water, and the product was obtained by lyophilization.

[0162] Step 4: Take 0.5 mL of diethylamine and 5 mL of N,N-dimethylformamide or N,N-dimethylacetamide and add them to a 10 mL reaction bottle. Stir for 10 minutes. Then add 50 mg of the above product at 60 oThe reaction was stirred at 40 °C for 7 hours. The resulting solution was precipitated with ether, and the precipitated solution was centrifuged to obtain a solid product. The solid product was purified by dialysis in purified water. The purified solid product was dried by freeze drying to obtain an aminated diblock polymer;

[0163] Step 5: Weigh 0.5 g of the aminated diblock polymer and dissolve it in 10 mL of dimethyl sulfoxide (or a corresponding solvent that can dissolve the compound), stir and dissolve it with a magnetic stirrer (500 rpm, 30 min) to completely dissolve it. Under ice bath conditions, slowly add hydrochloric acid (38%) or acetic acid, continuously monitor the pH of the reaction environment, and control the pH value at 1-3. After reaching the predetermined pH range, continue to stir the reaction for 1 h. After the reaction is completed, use anhydrous ether to precipitate and centrifuge the polymer. After repeating 2 to 5 times, vacuum dry the obtained solid sample to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

[0164] Example 179: Preparation of polymer A179 capable of assembling with proteins and hydrophobic small molecule drugs by RAFT reaction

[0165] The difference between the preparation method of embodiment 179 and embodiment 179 is that the same molar amount of 4-sodium azosulfonate calix[6]arene is used to replace the 4-sodium azosulfonate calix[4]arene in embodiment 12.

[0166] Comparative Example 1: This comparative example provides a commercially available embolic microsphere product Callispheres, which is blue in model and has a specification of 100-300 μm.

[0167] Test Example 1: Using polymers A1~A89, A91~A179 to improve the drug loading performance of the commercially available embolic microsphere product Callispheres;

[0168] The following tests were performed using polymer Ax (x ranged from 1 to 89, 91 to 179):

[0169] Dissolve 68 mg of sorafenib and 100 mg of polymer Ax in 1 mL of DMSO to form a clear and transparent solution. Add 50 mL of purified water and shake vigorously for 5 min to form a milky white translucent solution. Add 1 mL of drug-loaded embolic microspheres (Callispheres), continue shaking for 5 min, let stand for 10 min, and then continue shaking for 1 min. Repeat the above steps until 1 h. After the microspheres settle, take the upper solution for UV absorbance test at a wavelength of 280 nm.

[0170] Table 1 shows the performance test data of the commercially available embolic microsphere product Callispheres loaded with targeted drugs (sorafenib)

[0171] polymer Average drug loading rate after 10 min drug loading time Average drug loading rate at 30 min loading time Average drug loading rate after 60 min drug loading time A1 74% 86% 94% A2~A9 71% 84% 91% A10 61% 74% 80% A11 59% 68% 73% A12 42% 51% 57% A13 41% 52% 57% A14 76% 89% 96% A15~A23 72% 85% 93% A24 71% 86% 92% A25 52% 63% 68% A26 44% 53% 59% A27 45% 55% 60% A28 72% 83% 90% A29~A37 71% 82% 89% A38~A85 70% 83% 90% A86 63% 76% 82% A87 48% 59% 64% A88 46% 53% 61% A89 45% 52% 61% A91 74% 85% 92% A92~A99 75% 87% 93% A100 76% 89% 93% A101 72% 85% 90% A102 42% 51% 56% A103 43% 52% 58% A104 76% 88% 94% A105~A113 75% 86% 91% A114 79% 91% 98% A115 69% 81% 89% A116 45% 54% 61% A117 46% 54% 61% A118 75% 87% 92% A119~A127 74% 88% 93% A128~A175 72% 87% 93% A176 78% 90% 94% A177 69% 81% 87% A178 47% 52% 63% A179 46% 52% 62% Comparative Example 1 1% 2% 4%

[0172] Test Example 2:

[0173] Using polymers A1~A89, A91~A179 to improve the drug loading performance of the commercially available embolic microsphere product Callispheres;

[0174] The following tests were performed using polymer Ax (x ranged from 1 to 89, 91 to 179):

[0175] Dissolve 62 mg of lenvatinib and 100 mg of polymer Ax in 1 mL of DMSO to form a clear and transparent solution. Add 50 mL of purified water and shake vigorously for 5 min to form a milky white translucent solution. Add 1 mL of drug-loaded embolic microspheres (Callispheres), continue shaking for 5 min, let stand for 10 min, and continue shaking for 1 min. Repeat the above steps until 1 h. After the microspheres settle, take the upper solution for UV absorbance test at a wavelength of 280 nm.

[0176] Table 2 shows the performance test data of the commercially available embolic microsphere product Callispheres loaded with targeted drugs (lenvatinib)

[0177] polymer Average drug loading rate after 10 min drug loading time Average drug loading rate at 30 min loading time Average drug loading rate after 60 min drug loading time A1 52% 65% 71% A2~A9 50% 61% 68% A10 43% 55% 62% A11 36% 47% 51% A12 30% 35% 38% A13 32% 36% 39% A14 53% 65% 72% A15~A23 53% 63% 74% A24 51% 63% 68% A25 34% 44% 48% A26 23% 31% 36% A27 25% 35% 38% A28 50% 61% 70% A29~A37 52% 61% 67% A38~A85 50% 64% 69% A86 43% 56% 62% A87 24% 36% 41% A88 29% 38% 45% A89 30% 35% 44% A91 54% 62% 67% A92~A99 53% 64% 70% A100 51% 64% 74% A101 52% 66% 70% A102 33% 42% 52% A103 30% 40% 50% A104 54% 64% 72% A105~A113 53% 66% 71% A114 62% 74% 79% A115 44% 58% 66% A116 36% 48% 59% A117 35% 50% 62% A118 55% 68% 72% A119~A127 51% 65% 70% A128~A175 50% 65% 72% A176 58% 69% 74% A177 48% 59% 65% A178 32% 44% 51% A179 33% 46% 53% Comparative Example 1 1% 1% 2%

[0178] Test Example 3:

[0179] Using polymers A1~A89, A91~A179 to improve the drug loading performance of the commercially available embolic microsphere product Callispheres;

[0180] The following tests were carried out in sequence using polymers A1~A89 and A91~A179:

[0181] Take 1 mL of atelizumab injection (containing 60 mg of atelizumab), add 50 mg of polymer Ax (x ranges from 1 to 89, 91 to 179) to dissolve, shake vigorously for 5 min to form a clear and transparent solution. Add 1 mL of drug-loaded embolic microspheres (Callispheres), continue shaking for 5 min, let stand for 10 min, and then continue shaking for 1 min. Repeat the above steps until 1 h. After the microspheres settle, take the upper solution for UV absorbance test at a wavelength of 280 nm.

[0182] Table 3 shows the performance test data of the commercially available embolic microsphere product Callispheres loaded with immune drug (atelizumab)

[0183] polymer Average drug loading rate after 10 min drug loading time Average drug loading rate at 30 min loading time Average drug loading rate after 60 min drug loading time A1 41% 53% 60% A2~A9 34% 52% 63% A10 43% 55% 62% A11 34% 44% 52% A12 27% 31% 35% A13 30% 35% 38% A14 49% 62% 69% A15~A23 50% 58% 72% A24 52% 61% 65% A25 32% 46% 51% A26 19% 28% 32% A27 22% 30% 36% A28 48% 62% 74% A29~A37 44% 59% 68% A38~A85 51% 68% 72% A86 39% 51% 59% A87 27% 40% 48% A88 21% 30% 34% A89 22% 33% 38% A91 38% 47% 55% A92~A99 46% 55% 63% A100 54% 64% 71% A101 48% 61% 68% A102 30% 40% 45% A103 30% 39% 45% A104 51% 59% 62% A105~A113 46% 54% 61% A114 58% 66% 70% A115 44% 55% 60% A116 30% 41% 48% A117 30% 40% 48% A118 50% 62% 70% A119~A127 42% 54% 64% A128~A175 44% 56% 64% A176 50% 61% 67% A177 42% 59% 62% A178 27% 39% 44% A179 30% 45% 48 / % Comparative Example 1 5% 5% 6%

[0184] Comparative Example 2: This comparative example provides a commercially available drug-loaded embolic microsphere DC Bead, which is blue in model and has a specification of 100-300 μm.

[0185] Test Example 1: Using polymers A1~A89, A91~A179 to improve the drug loading performance of commercially available embolic microsphere product DC Bead;

[0186] The following tests were carried out in sequence using polymers A1~A89 and A91~A179:

[0187] Take 68 mg of sorafenib and 100 mg of polymer Ax (x ranges from 1 to 89, 91 to 179) and dissolve them in 1 mL of DMSO to form a clear and transparent solution. Add 50 mL of purified water and shake vigorously for 5 min to form a milky white translucent solution. Add 1 mL of commercially available drug-loaded embolic microspheres (DC Beads), continue shaking for 5 min, let stand for 10 min, and then continue shaking for 1 min. Repeat the above steps until 1 h. After the microspheres settle, take the upper solution for UV absorbance test at a wavelength of 280 nm.

[0188] Table 4 shows the performance test data of commercially available drug-loaded embolic microspheres (DC Bead) loaded with targeted drugs (sorafenib)

[0189] polymer Average drug loading rate after 10 min drug loading time Average drug loading rate at 30 min loading time Average drug loading rate after 60 min drug loading time A1 72% 83% 91% A2~A9 68% 79% 88% A10 63% 73% 82% A11 57% 69% 76% A12 44% 52% 59% A13 45% 53% 62% A14 72% 85% 94% A15~A23 73% 87% 92% A24 68% 85% 90% A25 53% 65% 67% A26 41% 50% 56% A27 42% 52% 58% A28 74% 84% 93% A29~A37 72% 84% 91% A38~A85 67% 82% 92% A86 60% 74% 84% A87 42% 56% 65% A88 44% 51% 60% A89 45% 53% 63% A91 74% 86% 95% A92~A99 71% 86% 94% A100 77% 92% 96% A101 70% 81% 90% A102 44% 56% 61% A103 45% 60% 65% A104 77% 90% 96% A105~A113 72% 84% 92% A114 77% 90% 99% A115 71% 84% 92% A116 42% 51% 56% A117 40% 50% 54% A118 77% 90% 94% A119~A127 68% 84% 89% A128~A175 71% 86% 93% A176 80% 90% 95% A177 71% 83% 90% A178 42% 51% 60% A179 41% 52% 61% Comparative Example 1 2% 4% 5%

[0190] Test Example 2:

[0191] Using polymers A1~A89, A91~A179 to improve the drug loading performance test of commercially available drug-loaded embolic microspheres (DC Bead);

[0192] The following tests were performed using polymer Ax (x ranged from 1 to 89, 91 to 179):

[0193] Take 62 mg of lenvatinib and 100 mg of polymer Ax (x ranges from 1 to 89, 91 to 179) and dissolve them in 1 mL of DMSO to form a clear and transparent solution. Add 50 mL of purified water and shake vigorously for 5 min to form a milky white translucent solution. Add 1 mL of drug-loaded embolic microspheres (DC Beads), continue shaking for 5 min, let stand for 10 min, and continue shaking for 1 min. Repeat the above steps until 1 hour. After the microspheres settle, take the upper solution for UV absorbance test at a wavelength of 280 nm.

[0194] Table 5 shows the performance test data of commercially available drug-loaded embolic microspheres (DC Bead) loaded with targeted drugs (lenvatinib)

[0195] polymer Average drug loading rate after 10 min drug loading time Average drug loading rate at 30 min loading time Average drug loading rate after 60 min drug loading time A1 50% 61% 69% A2~A9 52% 64% 72% A10 44% 54% 65% A11 31% 46% 52% A12 36% 40% 46% A13 35% 39% 45% A14 54% 68% 78% A15~A23 54% 66% 77% A24 49% 63% 67% A25 36% 45% 47% A26 20% 29% 32% A27 22% 30% 35% A28 54% 62% 68% A29~A37 50% 62% 69% A38~A85 52% 67% 71% A86 40% 55% 63% A87 22% 34% 42% A88 25% 33% 39% A89 30% 35% 42% A91 56% 64% 70% A92~A99 55% 68% 74% A100 50% 61% 70% A101 50% 64% 68% A102 34% 41% 46% A103 34% 42% 48% A104 52% 66% 72% A105~A113 48% 60% 66% A114 64% 75% 83% A115 44% 59% 68% A116 34% 46% 58% A117 36% 48% 60% A118 56% 66% 74% A119~A127 48% 62% 69% A128~A175 52% 68% 75% A176 56% 66% 71% A177 48% 61% 69% A178 33% 42% 46% A179 30% 40% 45% Comparative Example 1 1% 2% 2%

[0196] Test Example 3:

[0197] Using polymers A1~A89, A91~A179 to improve the drug loading performance test of commercially available drug-loaded embolic microspheres (DC Bead);

[0198] The following tests were carried out in sequence using polymers A1~A89 and A91~A179:

[0199] Take 1 mL of atelizumab injection (containing 60 mg of atelizumab), add 50 mg of polymer Ax (x ranges from 1 to 89, 91 to 179) to dissolve, shake vigorously for 5 min to form a clear and transparent solution. Add 1 mL of commercially available drug-loaded embolic microspheres (DCBead), continue shaking for 5 min, let stand for 10 min, and continue shaking for 1 min. Repeat the above steps until 1 h. After the microspheres settle, take the upper solution for ultraviolet absorbance test at a wavelength of 280 nm.

[0200] Table 6 shows the performance test data of commercially available drug-loaded embolic microspheres (DC Bead) loaded with immune drugs (atelizumab)

[0201] polymer Average drug loading rate after 10 min drug loading time Average drug loading rate at 30 min loading time Average drug loading rate after 60 min drug loading time A1 39% 51% 58% A2~A9 32% 50% 60% A10 40% 51% 58% A11 30% 42% 50% A12 24% 29% 33% A13 26% 32% 38% A14 52% 65% 72% A15~A23 51% 60% 74% A24 48% 58% 64% A25 33% 45% 52% A26 16% 25% 29% A27 18% 26% 30% A28 50% 61% 72% A29~A37 46% 61% 71% A38~A85 49% 67% 73% A86 35% 50% 56% A87 25% 41% 47% A88 18% 28% 32% A89 16% 27% 30% A91 34% 47% 53% A92~A99 44% 56% 64% A100 53% 62% 70% A101 46% 62% 69% A102 28% 42% 48% A103 30% 40% 46% A104 48% 60% 65% A105~A113 43% 56% 62% A114 56% 67% 73% A115 44% 53% 57% A116 31% 44% 51% A117 30% 42% 50% A118 48% 63% 71% A119~A127 40% 52% 60% A128~A175 45% 56% 66% A176 45% 58% 64% A177 41% 54% 63% A178 23% 36% 42% A179 28% 39% 45% Comparative Example 1 4% 5% 6%

[0202] It can be seen from the above test examples that, compared with the comparative examples, the diblock polymers A1~A86, A88~A173 prepared in the examples and capable of assembling with hydrophobic small molecules and protein drugs can effectively load small molecule targeted drugs, macromolecular targeted drugs and macromolecular immune drugs with existing drug-loaded microspheres Callispheres, DC Beads, etc. by combining with hydrophobic small molecule targeted drugs, macromolecular targeted drugs and macromolecular immune drugs through host-guest self-assembly. The reason is that:

[0203] For hydrophobic small molecule drugs such as sorafenib and lenvatinib, in DMSO solution, the diblock polymer and hydrophobic small molecule drugs prepared by the present invention are both in a dissolved state. When water is added to the system, the insoluble drug quickly assembles with the cavity-containing molecules in the diblock polymer, and after assembly, the assembly blocks are made hydrophobic, and finally form a vesicle structure in the aqueous solution under the action of the cationic blocks. The outer layer of the vesicle structure has a large number of hydrophilic amino cations, which interact with the negatively charged sulfonic acid groups on the microsphere polymer skeleton, so that the microspheres can load hydrophobic small molecule drugs such as sorafenib and lenvatinib;

[0204] For water-soluble protein drugs such as atelizumab, the diblock polymer is attached to the protein surface through multiple cavity structures in the cyclodextrin block in the aqueous solution, thereby forming a layer of diblock polymer on the surface of the protein molecule. The cationic groups in the polymer interact with the negatively charged sulfonic acid groups on the microsphere polymer skeleton, so that the microspheres can load protein drugs such as atelizumab.

[0205] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0206] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A block polymer that can be assembled with proteins and hydrophobic small molecule drugs, characterized in that: It has any one of the following structural formulas: ， ， ， , , , wherein the R group is a functional group containing cyclodextrin or calixarene, n is the number of ammonium blocks, m is the number of cavity-containing molecular blocks, and n and m are each independently selected from an integer greater than or equal to 2.

2. The block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 1, characterized in that: The polymer is selected from the following structures: ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， 。 3. A method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs as claimed in any one of claims 1 or 2, characterized in that: include: Any one of an ammonium functional group-containing monomer, an amine functional group-containing monomer and an alkylene oxide functional group-containing monomer is selected as a main material, and a polymer intermediate having a grafting site is obtained through a first ATRP reaction or a RAFT reaction, and the polymer intermediate is subjected to a second ATRP reaction or a RAFT reaction to graft a main molecule polymer containing a cavity structure or a polymer containing hydroxyl groups and ester groups, and the polymer containing hydroxyl groups and ester groups is modified by a cavity-containing organic molecule to obtain a diblock polymer with a cavity functional group, wherein the main molecule polymer containing a cavity structure is any one of poly(propyl methacrylate)-2-hydroxy-3-methylamino-α-cyclodextrin, poly(propyl methacrylate)-2-hydroxy-3-methylamino-β-cyclodextrin and poly(propyl methacrylate)-2-hydroxy-3-methylamino-γ-cyclodextrin, and the cavity-containing organic molecule is modified from 4-sodium azosulfonate calix[4]arene or 4-sodium azosulfonate calix[6]arene; When the main material is a monomer containing an ammonium functional group, the diblock polymer with a cavity functional group is the polymer that can be assembled with proteins and hydrophobic small molecule drugs; When the main material is a monomer containing an amine functional group, the diblock polymer with a cavity functional group is cationized with an amine group to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs; When the main material is a monomer containing an alkylene oxide functional group, the diblock polymer with a cavity functional group is organically modified and grafted with an organic molecule containing an amine functional group to obtain an aminated polymer, and the amine group is cationized to obtain the polymer that can be assembled with proteins and hydrophobic small molecule drugs.

4. The method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 3, characterized in that: The ammonium functional group-containing monomer includes any one of N-(3-aminopropyl)methacrylamide hydrochloride, 2-aminoethyl methacrylate hydrochloride, allylamine hydrochloride, 2-methylallylamine hydrochloride, O-allylhydroxylamine hydrochloride, 3-butenylamine hydrochloride, 3-methylbut-2-en-1-amine hydrochloride, 2-allylaniline hydrochloride, and N-(3-aminopropyl)methacrylamide hydrochloride.

5. The method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 3, characterized in that: The amino functional group-containing monomer includes any one of 2-methylallylamine, acrylamide, N-(2-amino-2-oxoethyl)acrylamide, 2-buteneamide, 4-aminostilbene, 3-aminostyrene, 4-aminostyrene, methacrylamide, 2-methylallylamine, and 4-vinylbenzylamine.

6. The method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 3, characterized in that: The epoxy functional group-containing monomer includes any one of glycidyl methacrylate, allyl glycidyl ether, 3,4-epoxy-1-butene, (3,4-epoxycyclohexyl) methyl acrylate, 1,2-epoxy-9-decene, 1,2-epoxy-5-hexene, 1,2-epoxy-4-vinylcyclohexane, glycidyl acrylate, and glycidyl methacrylate.

7. The method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 3, characterized in that: The main molecular polymer containing the cavity structure is polymerized by a monomer containing a cavity functional group, and the monomer containing a cavity functional group includes any one of propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, propyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin, and propyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin.

8. The method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 7, characterized in that: The method for preparing the monomer containing a cavity functional group comprises: Any one of α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin is selected as a raw material of a monomer containing a cavity functional group and dissolved in an alkaline solution, and a modifier, toluenesulfonyl chloride, is added to react for several hours, and then the filtrate is collected and subjected to reduced pressure distillation and sedimentation to obtain a solid sample I; The solid sample I and the modifier 1,3-propylenediamine are mixed and reacted, and then distilled and precipitated under reduced pressure to obtain a solid sample II; Solid sample II and modifier glycidyl methacrylate are dissolved in an organic solvent, nitrogen is introduced, the temperature is increased, reaction is performed, and then precipitation is performed to obtain a solid sample, which is propyl methacrylate-2-hydroxy-3-methylamino-α-cyclodextrin, propyl methacrylate-2-hydroxy-3-methylamino-β-cyclodextrin or propyl methacrylate-2-hydroxy-3-methylamino-γ-cyclodextrin corresponding to the raw material.

9. The method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 3, characterized in that: The polymer containing hydroxyl and ester groups is obtained by polymerizing any one of hydroxyethyl methacrylate, allyl 4-hydroxybenzoate, 6-(allyloxycarbonylamino)-1-hexanol, 3-hexenol lactate, hydroxyethyl acrylate, 6-hydroxyhexyl methacrylate, and hydroxypropyl acrylate.

10. The method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 3, characterized in that: The method for preparing the cavity-containing organic molecule comprises: selecting 4-azosulfonic acid sodium calix[4]arene or 4-azosulfonic acid sodium calix[6]arene, dissolving it in an alkaline solution, adding a modifier epibromopropane, mixing the mixture, and then precipitating the mixture with ether to obtain the cavity-containing organic molecule.

11. The method for preparing a block polymer that can be assembled with proteins and hydrophobic small molecule drugs according to claim 7 or 10, characterized in that: The molar ratio of any one of the monomers containing ammonium functional groups, amine functional groups and alkylene oxide functional groups to the monomer containing cavity functional groups or the organic molecule containing cavity is (0.25-4):

1.

12. A method for improving the drug loading performance of drug-loaded microspheres, characterized in that: The drug-loaded microspheres are added into a mixed solution of the drug to be loaded and the block polymer as claimed in any one of claims 1 or 2.

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