Random polymer and preparation method thereof, nano adjuvant and nano vaccine

Abstract

The invention discloses a random polymer and a preparation method thereof, a nano adjuvant and a nano vaccine, and relates to the field of biomedicine. The random polymer disclosed by the invention has a hydrophilic structure, a first repetitive unit and a second repetitive unit, and can be self-assembled into nano particles, when the random polymer is used as a nano adjuvant, the nano particles are protonated in an antigen presenting cell endosome acidic environment, positive ions and hydrophobic structures in the random polymer are exposed, and the positive ions and the hydrophobic structures in the random polymer are exposed. And the lipid membrane is destroyed in a membrane disturbance manner, so that the endosome lipid membrane is quickly penetrated, and efficient antigen cytoplasm delivery is realized. Experimental results show that when the random polymer provided by the invention is used as a nano adjuvant, antigen-presenting cells can be effectively activated, the antigen cross-presenting efficiency is improved, and cellular immune response is stimulated.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a random polymer and a preparation method thereof, a nano adjuvant and a nano vaccine. Background Art

[0002] Vaccine injection is a fast, effective and low-cost immunotherapy. Vaccine adjuvant is an important ingredient in vaccines. It can enhance the intensity, breadth and durability of the immune response, and solve the problems of inability to effectively stimulate the body's immune response and unsatisfactory treatment effect caused by the low immunogenicity of the antigens in the vaccine.

[0003] In recent years, the rapid development of nanotechnology has provided new ideas for upgrading vaccine adjuvants. Nanoadjuvants prepared by nanotechnology or combining nanomaterials have the characteristics of good biocompatibility, good targeting, safety and high efficiency. However, current nanoadjuvants still have problems such as low efficiency and inability to induce effective immune responses. Therefore, it is necessary to develop new nanoadjuvants to induce sufficiently strong antigen-specific immune responses. Summary of the invention

[0004] In view of this, the present invention provides a random polymer and a preparation method thereof, a nanoadjuvant and a nanovaccine. The random polymer provided by the present invention can effectively enhance the antigen-specific immune response when used as a nanoadjuvant, increase the level of antigen-specific T cells in the body, and help the vaccine to exert its effect.

[0005] The present invention provides a random polymer comprising a hydrophilic structure represented by formula (I), a first repeating unit represented by formula (II) and a second repeating unit represented by formula (III).

[0006]

[0007] wherein n is selected from an integer of 2 to 500, preferably an integer of 30 to 300, more preferably an integer of 50 to 200; m is selected from an integer of 1 to 20, preferably an integer of 1 to 10, more preferably an integer of 1 to 5; the molar ratio of the first repeating unit to the second repeating unit is (1 to 150):(1 to 150), preferably 0.1 to 63:1, more preferably 0.8 to 40:1;

[0008] R5 is selected from C1~C 30 Alkyl, C1~C 30 Alkyl, C2~C 30 Alkenyl, C2-C 30 Alkenyl, C2~C 30 Alkynyl, C2-C 30 Alkynyl, C3~C30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C3~C 30 cycloalkenyl, C3~C 30 Cycloalkenyl, C2~C 30 heterocyclic alkenyl, C2-C 30 Heterocycloalkenyl, C3~C 30 cycloalkynyl, C3~C 30 Cycloalkynyl, C2~C 30 heterocycloalkynyl, C2-C 30 Heterocycloalkynyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 Alkenyl, C2~C 30 Alkynyl, C2~C 30At least one of an amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl group;

[0009] R2 and R3 are independently selected from hydrogen, C1-C 30 Alkyl, C1~C 30 Alkyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 at least one of amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl; or R2 and R3 and the N atom to which they are connected together form a 3-30 membered heterocyclic ring;

[0010] R1 and R4 are independently selected from hydrogen, C1-C 30 Alkyl, C1~C 30Alkyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 At least one of amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl groups.

[0011] Specifically, the random polymer of the present invention may have the following structure:

[0012]

[0013] wherein x and y are independently selected from integers ranging from 2 to 300;

[0014] R6 is selected from hydrogen, C1~C 30 Alkyl, C1~C 30 Alkyl, C3~C 30cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 At least one of an amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl group;

[0015] The selection ranges of m, n and R1 to R5 are the same as those described above, and the present invention will not be repeated here.

[0016] The present invention provides a random polymer having a hydrophilic structure, a first repeating unit and a second repeating unit. Under a neutral pH condition, the random polymer can be assembled into nanoparticles. When used as a nanoadjuvant, it is protonated in the acidic environment of the endosome of an antigen presenting cell, exposing the cations and hydrophobic structures in the random polymer, destroying the lipid membrane in a membrane perturbation manner, thereby quickly passing through the lipid membrane of the endosome, achieving efficient antigen cytoplasmic delivery, and enhancing antigen cross-presentation and specific immune response.

[0017] In some specific implementations, the R5 is selected from C1 to C 25 Alkyl, C1~C 25 Alkyl, C2~C 25 Alkenyl, C2-C 25 Alkenyl, C2~C 25 Alkynyl, C2-C 25 Alkynyl, C3~C 25 cycloalkyl, C3~C 25 Cycloalkyl, C3~C 25 cycloalkenyl, C3~C 25 Cycloalkenyl, C3~C 25 cycloalkynyl, C3~C 25 Cycloalkynyl, C6~C 25 C6~C 25 The aromatic group, the substituent is selected from C1 to C 25 Alkyl, C2~C 25 Alkenyl, C2~C 25 Alkynyl and C6~C 25 At least one of the aromatic groups.

[0018] In some specific implementations, the R5 is preferably C1 to C 20 Alkyl, C1~C 20 Alkyl, C2~C 20 or at least one substituent substituted with C2-C 20 The substituent is selected from C1 to C 20 Alkyl, C2~C 20 Alkenyl, C3~C 20 Cycloalkyl or C6~C 20 An aromatic group; more preferably the following structure:

[0019]

[0020] Wherein, f is selected from an integer of 0 to 20;

[0021] R d Selected from hydrogen, C2~C 20 Alkenyl, C3~C 20 Cycloalkyl or C6~C 20 Aromatic base.

[0022] In some specific implementations, R2 and R3 are independently selected from C1-C5 alkyl groups or C1-C5 alkyl groups substituted with at least one substituent, wherein the substituent is selected from at least one of amino, hydroxyl, carboxyl, cyano and thiol; or R2, R3 and the N atom where they are located together form a ring having the following structure:

[0023]

[0024] wherein a, b, c, d and e are independently selected from integers of 1 to 5;

[0025] R a and R b Independently selected from hydrogen or C1-C5 alkyl;

[0026] —X—is selected from —O—,

[0027] R c Selected from hydrogen or C1-C5 alkyl.

[0028] In some specific implementations, the R2, R3 and the N atom to which they are commonly connected are preferably a ring having the following structure:

[0029]

[0030] Here, a is selected from integers of 1 to 5.

[0031] In some specific implementations, the R2 and R3 are more preferably formed together with the nitrogen atom where they are located to form a ring having the following structure:

[0032]

[0033] In some specific implementations, R1 and R4 are independently selected from hydrogen, C1-C 25 Alkyl, C3~C 25 Cycloalkyl, C6~C 25 Aromatic or C6~C 25 The heteroaryl group is preferably C1 to C 20 The alkyl group is more preferably a methyl group.

[0034] In some specific implementations, the random polymer has the following structure:

[0035]

[0036] wherein x and y are independently selected from integers ranging from 2 to 300;

[0037] f is an integer selected from 0 to 20;

[0038] Rd is selected from hydrogen or phenyl.

[0039] In some specific implementations, the number average molecular weight of the random polymer is 1,000 to 200,000, preferably 2,000 to 150,000, and more preferably 3,500 to 100,000.

[0040] The present invention also provides a method for preparing a random polymer, comprising:

[0041] A macromolecular chain transfer agent having a structure of formula (IV), a monomer having a structure of formula (V), a monomer having a structure of formula (VI) and an initiator undergo a polymerization reaction to obtain the random polymer;

[0042]

[0043] wherein n is selected from an integer of 2 to 500; m is selected from an integer of 1 to 20;

[0044] R5 is selected from C1~C 30 Alkyl, C1~C 30 Alkyl, C2~C 30 Alkenyl, C2-C 30 Alkenyl, C2~C 30 Alkynyl, C2-C 30 Alkynyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C3~C 30 cycloalkenyl, C3~C 30 Cycloalkenyl, C2~C 30 heterocyclic alkenyl, C2-C 30 Heterocycloalkenyl, C3~C 30 cycloalkynyl, C3~C 30 Cycloalkynyl, C2~C 30 heterocycloalkynyl, C2-C 30 Heterocycloalkynyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 Alkenyl, C2~C 30 Alkynyl, C2~C 30 At least one of an amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl group;

[0045] R2 and R3 are independently selected from hydrogen, C1-C 30 Alkyl, C1~C 30 Alkyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 at least one of amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl; or R2 and R3 and the N atom to which they are connected together form a 3-30 membered heterocyclic ring;

[0046] R1, R4 and R6 are independently selected from hydrogen, C1-C 30 Alkyl, C1~C 30 Alkyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 At least one of amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl groups.

[0047] The present invention uses a hydrophilic macromolecular chain transfer agent having a structure of formula (IV), a (meth)acrylate monomer containing a tertiary amine having a structure of formula (V), and a (meth)acrylate monomer containing a hydrophobic structure having a structure of formula (VI) as raw materials, and under the action of an initiator, a reversible addition-fragmentation chain transfer polymerization (RAFT) reaction is performed to obtain a random polymer. Specifically, the random polymer can be synthesized according to the route shown in the following scheme:

[0048]

[0049] In the present invention, the hydrophilic macromolecular chain transfer agent having the structure of formula (IV) provides easily breakable chemical bonds, so that an exchange reaction occurs between the polymerized monomer and the macromolecular chain transfer agent during the reaction, thereby interrupting or extending the random polymer chain, thereby achieving regulation of the random polymer molecular chain and molecular weight distribution.

[0050] In some specific implementations, the hydrophilic macromolecular chain transfer agent having the structure of formula (IV) is selected from the following structures:

[0051]

[0052] Among them, g is selected from an integer of 0 to 20, preferably an integer of 1 to 15, and more preferably an integer of 1 to 11; n is selected from an integer of 2 to 500, preferably an integer of 30 to 300, and more preferably an integer of 50 to 200.

[0053] In some specific implementations, the hydrophilic macromolecular chain transfer agent having the structure of formula (IV) is synthesized by the following method:

[0054] A carboxylic acid chain transfer agent, monomethyl ether polyethylene glycol, 4-dimethylaminopyridine (DMAP) and N,N'-diisopropylcarbodiimide (DIC) react in a solvent to obtain a hydrophilic macromolecular chain transfer agent having a structure of formula (IV).

[0055] In some specific implementations, the solvent is selected from dichloromethane (DCM), preferably anhydrous DCM. In some specific implementations, the carboxylic acid chain transfer agent is selected from 4-cyano-4-(dodecylsulfanylthiocarbonyl)sulfanylpentanoic acid (CDTPA), 4-cyano-4-(n-hexylsulfanylthiocarbonyl)sulfanylpentanoic acid (CHTPA), 4-cyano-4-(ethylsulfanylthiocarbonyl)sulfanylpentanoic acid (CETPA) or 4-cyano-4-(phenylcarbonylthiothiothiothio)pentanoic acid (CPTPA). In some specific implementations, the reaction temperature is 0 to 50°C, preferably 10 to 40°C, and more preferably 20 to 30°C. In some specific implementations, the reaction time is 6 to 72h, preferably 15 to 60h, and more preferably 24 to 48h. In some specific implementations, the molar ratio of the monomethyl ether polyethylene glycol to the carboxylic acid chain transfer agent is 1:2-5, preferably 1:3-4. In some specific implementations, the molar ratio of the monomethyl ether polyethylene glycol to DMAP is 1:0.1-0.5, preferably 1:0.2-0.4, more preferably 1:0.2-0.3. In some specific implementations, the molar ratio of the monomethyl ether polyethylene glycol to DIC is 1:3-8, preferably 1:4-7, more preferably 1:4-6.

[0056] After obtaining the hydrophilic macromolecular chain transfer agent having the structure of formula (IV), it is preferably purified to obtain a pure compound. In some specific implementations, the purification includes sedimenting the reaction product through ice anhydrous ether, and obtaining a crude product by suction filtration, and then dissolving the crude product with deionized water and transferring it to a dialysis bag (3500MWCO) for dialysis for 72 hours, and obtaining a pure hydrophilic macromolecular chain transfer agent having the structure of formula (IV) after freeze-drying. The present invention has no particular limitation on the purification method.

[0057] In the present invention, the monomer having the structure of formula (V) provides a hydrophobic group containing a tertiary amine structure for the random polymer, which enables the random polymer to pass through the lipid membrane through the hydrophobic effect, thereby enhancing the transmembrane delivery ability.

[0058] In some specific implementations, the monomer having the structure of formula (V) is synthesized by the following method:

[0059] The tertiary amine compound, triethylamine (TEA) and the inhibitor are dissolved in DCM, cooled to 0°C in an ice bath, and the DCM solution of the acyl chloride compound is slowly added dropwise, and then the mixed solution is transferred to room temperature and stirred for 6 to 12 hours. After the reaction is completed, the insoluble impurities are removed to obtain a filtrate, which is purified to obtain a monomer having a structure of formula (V). Specifically, the monomer having a structure of formula (V) can be synthesized according to the route shown in the following scheme:

[0060]

[0061] Among them, the selection ranges of R1, R2, R3 and m are the same as those described above, and the present invention will not be repeated here.

[0062] In some specific implementations, the DCM is preferably dry DCM. In some specific implementations, the tertiary amine compound is selected from N-(2-hydroxyethyl) hexamethylenediamine. In some specific implementations, the inhibitor is selected from hydroquinone. In some specific implementations, the acyl chloride compound is selected from acryloyl chloride or methacryloyl chloride. In some specific implementations, the purification includes concentrating the filtrate and purifying it by column chromatography or vacuum distillation, preferably vacuum distillation purification. The present invention has no particular restrictions on the purification method.

[0063] In some specific implementations, the molar ratio of the tertiary amine compound to the acyl chloride compound is 1:1-2, preferably 1:1-1.5, and more preferably 1:1-1.1. In some specific implementations, the molar ratio of the tertiary amine compound to TEA is 1:2-5, preferably 1:2-4, and more preferably 1:2-3. In some specific implementations, the molar ratio of the tertiary amine compound to the inhibitor is 1:0.005-0.02, preferably 1:0.008-0.02, and more preferably 1:0.01-0.02.

[0064] In the present invention, the monomer having the structure of formula (VI) provides a hydrophobic group for the random polymer. Such a structure can act as a detergent to rupture the lipid membrane of the endosomal membrane, thereby enhancing the ability of the random polymer to mediate the delivery of cargo to the cell cytoplasm, achieving efficient endosomal escape and antigen cytoplasmic delivery, and enhancing the ability of antigen cross-presentation.

[0065] In some specific implementations, the monomer having the structure of formula (VI) is synthesized by the following method:

[0066] The alcohol compound, TEA and inhibitor are dissolved in DCM, cooled to 0°C in an ice bath, and the DCM solution of the acyl chloride compound is slowly added dropwise, and then the mixed solution is transferred to room temperature and stirred for 6 to 12 hours. After the reaction is completed, the insoluble impurities are removed and the monomer having the structure of formula (VI) is obtained after purification. Specifically, the monomer having the structure of formula (VI) can be synthesized according to the route shown in the following scheme:

[0067]

[0068] The selection ranges of R4 and R5 are the same as those described above, and the present invention will not be repeated here.

[0069] In some specific implementations, the DCM is preferably dry DCM. In some specific implementations, the alcohol compound is selected from n-hexanol. In some specific implementations, the inhibitor is selected from hydroquinone. In some specific implementations, the acyl chloride compound is selected from acryloyl chloride or methacryloyl chloride. In some specific implementations, the purification includes concentrating the filtrate and then purifying it by column chromatography or vacuum distillation, preferably by vacuum distillation. The present invention has no particular restrictions on the purification method.

[0070] In some specific implementations, the molar ratio of the alcohol compound to the acyl chloride compound is 1:1-2, preferably 1:1-1.5, and more preferably 1:1-1.1. In some specific implementations, the molar ratio of the alcohol compound to TEA is 1:2-5, preferably 1:2-4, and more preferably 1:2-3. In some specific implementations, the molar ratio of the alcohol compound to the inhibitor is 1:0.005-0.02, preferably 1:0.008-0.02, and more preferably 1:0.01-0.02.

[0071] The present invention firstly dissolves a hydrophilic macromolecular chain transfer agent having a structure of formula (IV), a (meth)acrylate monomer containing a tertiary amine structure having a structure of formula (V), a (meth)acrylate monomer containing a hydrophobic group having a structure of formula (VI), and an initiator in a solvent, deoxygenates the reaction system, and causes a RAFT polymerization reaction under a sealed condition. After the reaction is completed, the reaction is quenched to obtain the random polymer.

[0072] In some specific implementations, the solvent of the RAFT polymerization reaction is one or more of dioxane, N,N-dimethylacetamide and N,N-dimethylformamide, preferably N,N-dimethylacetamide or N,N-dimethylformamide, more preferably N,N-dimethylacetamide. In some specific implementations, the initiator is selected from azobisisobutyronitrile (AIBN) or azobiscyanovaleric acid (V-501), preferably AIBN, more preferably AIBN after recrystallization. In some specific implementations, the deoxygenation operation includes deoxygenation using a high-purity inert gas, preferably nitrogen or argon, more preferably argon. The present invention has no special restrictions on the deoxygenation method. In some specific implementations, the temperature of the RAFT polymerization reaction is 35 to 70°C, preferably 40 to 70°C, more preferably 45 to 70°C. In some specific implementations, the time of the RAFT polymerization reaction is 12 to 48h, preferably 15 to 40h, more preferably 20 to 30h. In some specific implementations, the quenching reaction is to place the reaction system directly under low temperature after the reaction is completed. In some specific implementations, the low temperature is -10 to -50°C, preferably -10 to -40°C, and more preferably -10 to -30°C.

[0073] After obtaining the random polymer, the present invention preferably purifies it to obtain a pure random polymer. In some specific implementations, the purification includes placing the reaction solution in a dialysis bag, dialyzing with methanol and deionized water, and freeze-drying after the dialysis to obtain a pure random polymer. In some specific implementations, the molecular weight cutoff of the dialysis bag is 2500-4000MWCO, preferably 2800-3800MWCO, and more preferably 3200-3600MWCO. In some specific implementations, the dialysis time is 48-96h, preferably 60-84h, and more preferably 65-80h. The present invention has no particular restrictions on the specific method of purification.

[0074] In some specific implementations, the random polymer can also be obtained by atom transfer radical polymerization (ATRP), which will not be described in detail in the present invention.

[0075] The present invention provides a nanoparticle, comprising the random polymer. In some specific implementations, the nanoparticle has a particle size of 20 to 100 nm.

[0076] The present invention provides a nano adjuvant, comprising the above-mentioned random polymer or nanoparticle.

[0077] The present invention provides a method for preparing a nano adjuvant, comprising:

[0078] The random polymer is dissolved in a sodium dihydrogen phosphate solution to make the pH value of the solution reach 5.4, and then the sodium dihydrogen phosphate solution is added to make the pH value reach 7.4 to obtain a nano adjuvant.

[0079] In some specific implementations, the concentration of the sodium dihydrogen phosphate solution is 0.1-0.5M, preferably 0.15-0.3M, and more preferably 0.15-0.25M.

[0080] In some specific implementations, the nanoadjuvant can also be prepared by a nanoprecipitation method, as follows:

[0081] The random polymer is dissolved in N,N-dimethylformamide to obtain a solution with a concentration of 10 to 150 mg / mL, preferably 10 to 100 mg / mL, and more preferably 20 to 80 mg / mL. Deionized water is added under stirring, and the N,N-dimethylformamide is dialyzed in deionized water to obtain a nano adjuvant.

[0082] The present invention provides a nano vaccine, comprising the above-mentioned nano adjuvant, antigen and an adjuvant acceptable in the vaccine. In some specific implementations, the antigen includes a model antigen, a tumor-associated antigen, a viral antigen, a bacterial antigen, a malarial antigen, and the genetic material of the antigen such as RNA and DNA. In some specific implementations, the nano vaccine provided by the present invention can prevent or treat different disease models according to the different co-mixed antigens.

[0083] The present invention provides a random polymer having a hydrophilic structure, a first repeating unit and a second repeating unit. The structure of the polymer can be precisely controlled by controlling the type of monomers fed and the feed ratio. When used together with an antigen as a nanoadjuvant, the polymer can be rapidly protonated and respond to expose the cation and hydrophobic structure in the acidic endosomal environment of antigen presenting cells. As a detergent, the lipid membrane of the endosomal membrane is broken, and efficient endosomal escape and antigen cytoplasmic delivery are achieved, thereby enhancing the ability of antigen cross-presentation and enhancing the inherent immune response and adaptive immune response in the body. The random polymer provided by the present invention is of great significance to the development of polymer nanoadjuvants. BRIEF DESCRIPTION OF THE DRAWINGS

[0084] Figure 1 The random polymer P(N7-methyl) provided by the present invention 10 )~P(N7-A 40 ) structure and H NMR spectrum;

[0085] Figure 2 The random polymer P(N7-propylene) provided by the present invention 10 )~P(N7- 40 ) structure and H NMR spectrum;

[0086] Figure 3 The random polymer P(N7-hexane) provided by the present invention 10 )~P(N7-H 40 ) structure and H NMR spectrum;

[0087] Figure 4 The random polymer P(N7-benzyl 10 )~P(N7-benzyl 40 ) structure and H NMR spectrum;

[0088] Figure 5 The random polymer P(N7-octyl 10 )~P(N7- 40 ) structure and H NMR spectrum;

[0089] Figure 6 The random polymer P(N7-decyl) provided by the present invention 10 )~P(N7-decene 40 ) structure and H NMR spectrum;

[0090] Figure 7 The random polymer P(N7-12) provided by the present invention 10 )~P(N7-Twelve 40 ) structure and H NMR spectrum;

[0091] Figure 8 The random polymer P(N7-18) provided by the present invention 10 )~P(N7-Eighteen 40 ) structure and H NMR spectrum;

[0092] Fig. 9 The average particle size of the nanoparticles formed by the random polymer provided by the present invention;

[0093] Fig.10 The experimental results of specific immune response in mice after injection of the nano vaccine provided by the present invention;

[0094] Fig.11 It is the proportion of antigen-specific T cells in mice after injection of the nano vaccine provided by the present invention. DETAILED DESCRIPTION

[0095] It should be understood that the expression "one or more of..." includes each of the items recited after the expression individually and in various different combinations of two or more of the recited items, unless otherwise understood from the context and usage. The expression "and / or" in combination with three or more recited items should be understood to have the same meaning, unless otherwise understood from the context.

[0096] The use of the terms "comprising", "having" or "containing", including their grammatical synonyms, should generally be understood as open and non-restrictive, for example not excluding other unrecited elements or steps, unless otherwise specifically stated or otherwise understood from the context.

[0097] It should be understood that the order of steps or the order in which certain actions are performed is not important as long as the present invention remains operable. In addition, two or more steps or actions may be performed simultaneously.

[0098] The use of any and all examples or exemplary language, such as "for example" or "including", herein is intended only to better illustrate the invention and does not limit the scope of the invention unless otherwise claimed. No language in this specification should be construed as indicating that any non-claimed element is essential to the practice of the invention.

[0099] In addition, the numerical ranges and parameters used to define the present invention are approximate values, and the relevant values ​​in the specific embodiments have been presented as accurately as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in this disclosure are modified by "about". Here, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a specific value or range.

[0100] The present invention is further described below in conjunction with the following examples. The protection scope of the present invention is not limited by the following examples.

[0101] Example 1

[0102] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer methyl methacrylate (0.0216 g, 215.8 μmol), initiator azobisisobutyronitrile (AIBN) (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Use a soft rubber stopper to seal the single-necked flask, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-methyl) 10), a pure light yellow powder (0.471 g, 88.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:10. Figure 1 In the H NMR spectrum shown in the figure, the line corresponding to number 1 on the right is the random polymer P(N7- 10 ) H NMR spectrum; the peak labeled 1 in the structure shown is the hydrogen area peak corresponding to the hydrophobic polymerization unit, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chain and other polymerization units.

[0103]

[0104] Example 2

[0105] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer methyl methacrylate (0.0433 g, 430.6 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-methyl) 20 ), a pure light yellow powder (0.479 g, 86.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:20. Figure 1 In the H NMR spectrum shown in the figure, the line corresponding to number 2 on the right is the random polymer P(N7- 20 ) H NMR spectrum; the peak labeled 1 in the structure shown is the hydrogen area peak corresponding to the hydrophobic polymerization unit, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chain and other polymerization units.

[0106]

[0107] Example 3

[0108] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer methyl methacrylate (0.0647 g, 645.9 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-methyl) 30 ), a pure light yellow powder (0.503 g, 87.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:30. Figure 1 In the H NMR spectrum shown in the figure, the line corresponding to number 3 on the right is the random polymer P(N7- 30 ) H NMR spectrum; the peak labeled 1 in the structure shown is the hydrogen area peak corresponding to the hydrophobic polymerization unit, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chain and other polymerization units.

[0109]

[0110] Example 4

[0111] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer methyl methacrylate (0.0862 g, 861.1 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-methyl) 40), pure light yellow powder (0.510 g, 85.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:40. Figure 1 In the H NMR spectrum shown in the figure, the line corresponding to number 4 on the right is the random polymer P(N7- 40 ) H NMR spectrum; the peak labeled 1 in the structure shown is the hydrogen area peak corresponding to the hydrophobic polymerization unit, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chain and other polymerization units.

[0112]

[0113] Example 5

[0114] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer propyl methacrylate (0.0261 g, 203.6 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7- 10 ), a pure light yellow powder (0.448 g, 83.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:10. Figure 2 In the H NMR spectrum shown in the figure, the line corresponding to number 1 on the right is the random polymer P(N7- 10 ) H NMR spectrum; the peaks labeled 1 and 2 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0115]

[0116] Example 6

[0117] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer propyl methacrylate (0.0522 g, 407.2 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7- 20 ), a pure light yellow powder (0.458 g, 81.5%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:20. Figure 2 In the H NMR spectrum shown in the figure, the line corresponding to number 2 on the right is the random polymer P(N7- 20 ) H NMR spectrum; the peaks labeled 1 and 2 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0118]

[0119] Example 7

[0120] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer propyl methacrylate (0.0783 g, 610.8 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7- 30), a pure light yellow powder (0.458 g, 81.5%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:30. Figure 2 In the H NMR spectrum shown in the figure, the line corresponding to number 3 on the right is the random polymer P(N7- 30 ) H NMR spectrum; the peaks labeled 1 and 2 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0121]

[0122] Example 8

[0123] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer propyl methacrylate (0.1045 g, 814.4 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7- 40 ), a pure light yellow powder (0.507 g, 82.5%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:40. Figure 2 In the H NMR spectrum shown in the figure, the line corresponding to number 4 on the right is the random polymer P(N7- 40 ) H NMR spectrum; the peaks labeled 1 and 2 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0124]

[0125] Example 9

[0126] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-hexyl methacrylate (0.0341 g, 200.2 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-hexane) 10 ), a pure light yellow powder (0.438 g, 80.5%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:10. Figure 3 In the H NMR spectrum shown in the figure, the line corresponding to number 1 on the right is the random polymer P(N7-hexane 10 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0127]

[0128] Example 10

[0129] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-hexyl methacrylate (0.0681 g, 400.4 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-hexane) 20), a pure light yellow powder (0.471 g, 81.5%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:20. Figure 3 In the H NMR spectrum shown in the figure, the line corresponding to number 2 on the right is the random polymer P(N7-hexane 20 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0130]

[0131] Embodiment 11

[0132] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-hexyl methacrylate (0.1022 g, 600.6 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-hexane) 30 ), a pure light yellow powder (0.499 g, 81.5%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:30. Figure 3 In the H NMR spectrum shown in the figure, the line corresponding to number 3 on the right is the random polymer P(N7-hexane 30 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0133]

[0134] Example 12

[0135] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-hexyl methacrylate (0.1362 g, 800.8 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-hexane) 40 ), pure light yellow powder (0.520 g, 80.5%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:40. Figure 3 In the H NMR spectrum shown in the figure, the line corresponding to number 4 on the right is the random polymer P(N7-hexane 40 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0136]

[0137] Example 13

[0138] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer benzyl methacrylate (0.0339 g, 192.4 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-benzyl) 10), a pure light yellow powder (0.455 g, 83.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:10. Figure 4 In the H NMR spectrum shown in the figure, the line corresponding to number 1 on the right is the random polymer P(N7-benzyl 10 ) H NMR spectrum; the peaks labeled 1 and 2 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0139]

[0140] Embodiment 14

[0141] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer benzyl methacrylate (0.0679 g, 385.4 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-benzyl) 20 ), a pure light yellow powder (0.466 g, 80.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:20. Figure 4 In the H NMR spectrum shown in the figure, the line corresponding to number 2 on the right is the random polymer P(N7-benzyl 20 ) H NMR spectrum; the peaks labeled 1 and 2 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0142]

[0143] Embodiment 15

[0144] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer benzyl methacrylate (0.1017 g, 577.2 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-benzyl) 30 ), a pure light yellow powder (0.497 g, 81.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:30. Figure 4 In the H NMR spectrum shown in the figure, the line corresponding to number 3 on the right is the random polymer P(N7-benzyl 30 ) H NMR spectrum; the peaks labeled 1 and 2 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0145]

[0146] Example 16

[0147] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer benzyl methacrylate (0.1356 g, 769.6 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-benzyl) 40), a pure light yellow powder (0.518 g, 80.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:40. Figure 4 In the H NMR spectrum shown in the figure, the line corresponding to number 4 on the right is the random polymer P(N7-benzyl 40 ) H NMR spectrum; the peaks labeled 1 and 2 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks labeled a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0148]

[0149] Embodiment 17

[0150] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-octyl methacrylate (0.0383 g, 193.1 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500 MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7- 10 ), a pure light yellow powder (0.462 g, 84.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:10. Figure 5 In the H NMR spectrum shown in the figure, the line corresponding to number 1 on the right is the random polymer P(N7- 10 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0151]

[0152] Embodiment 18

[0153] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-octyl methacrylate (0.0767 g, 386.2 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500 MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7- 20 ), a pure light yellow powder (0.489 g, 83.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:20. Figure 5 In the H NMR spectrum shown in the figure, the line corresponding to number 2 on the right is the random polymer P(N7- 20 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0154]

[0155] Embodiment 19

[0156] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-octyl methacrylate (0.1150 g, 579.3 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500 MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7- 30), pure light yellow powder (0.521 g, 83.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:30. Figure 5 In the H NMR spectrum shown in the figure, the line corresponding to number 3 on the right is the random polymer P(N7- 30 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0157]

[0158] Embodiment 20

[0159] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-octyl methacrylate (0.1534 g, 772.4 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500 MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7- 40 ), pure light yellow powder (0.546 g, 82.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:40. Figure 5 In the H NMR spectrum shown in the figure, the line corresponding to number 4 on the right is the random polymer P(N7- 40 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0160]

[0161] Embodiment 21

[0162] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-decyl methacrylate (0.0435 g, 192.1 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-decyl 10 ), a pure light yellow powder (0.472 g, 85.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:10. Figure 6 In the H NMR spectrum shown in the figure, the line corresponding to number 1 on the right is the random polymer P(N7-decene 10 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0163]

[0164] Embodiment 22

[0165] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-decyl methacrylate (0.0870 g, 384.2 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-decyl 20), pure light yellow powder (0.505 g, 84.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:20. Figure 6 In the H NMR spectrum shown in the figure, the line corresponding to number 2 on the right is the random polymer P(N7-decene 20 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0166]

[0167] Embodiment 23

[0168] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-decyl methacrylate (0.1304 g, 576.3 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-decyl 30 ), a pure light yellow powder (0.535 g, 83.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:30. Figure 6 In the H NMR spectrum shown in the figure, the line corresponding to number 3 on the right is the random polymer P(N7-decene 30 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0169]

[0170] Embodiment 24

[0171] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-decyl methacrylate (0.1739 g, 768.4 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-decyl 40 ), a pure light yellow powder (0.558 g, 81.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:40. Figure 6 In the H NMR spectrum shown in the figure, the line corresponding to number 4 on the right is the random polymer P(N7-decene 40 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0172]

[0173] Embodiment 25

[0174] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-dodecyl methacrylate (0.0435 g, 192.1 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-12 10), a pure light yellow powder (0.446 g, 80.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:10. Figure 7 In the H NMR spectrum shown in the figure, the line corresponding to number 1 on the right is the random polymer P(N7-12 10 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0175]

[0176] Embodiment 26

[0177] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-dodecyl methacrylate (0.0870 g, 384.2 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-12 20 ), a pure light yellow powder (0.487 g, 81.5%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:20. Figure 7 In the H NMR spectrum shown in the figure, the line corresponding to number 2 on the right is the random polymer P(N7-12 20 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0178]

[0179] Embodiment 27

[0180] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-dodecyl methacrylate (0.1304 g, 576.3 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-12 30 ), a pure light yellow powder (0.527 g, 82.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:30. Figure 7 In the H NMR spectrum shown in the figure, the line corresponding to number 3 on the right is the random polymer P(N7-12 30 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0181]

[0182] Embodiment 28

[0183] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer n-dodecyl methacrylate (0.1739 g, 768.1 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-12 40), a pure light yellow powder (0.549 g, 80.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:40. Figure 7 In the H NMR spectrum shown in the figure, the line corresponding to number 4 on the right is the random polymer P(N7-12 40 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0184]

[0185] Embodiment 29

[0186] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer methacrylate n-octadecyl (0.0627 g, 185.2 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500 MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-18 10 ), a pure light yellow powder (0.460 g, 80.3%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:10. Figure 8 In the H NMR spectrum shown in the figure, the line corresponding to number 1 on the right is the random polymer P(N7-18 10 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0187]

[0188] Embodiment 30

[0189] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer methacrylate n-octadecyl (0.1254 g, 370.4 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500 MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-18 20 ), pure light yellow powder (0.518 g, 81.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:20. Figure 8 In the H NMR spectrum shown in the figure, the line corresponding to number 2 on the right is the random polymer P(N7-18 20 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0190]

[0191] Embodiment 31

[0192] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer methacrylate n-octadecyl (0.1881 g, 555.5 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500 MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-18 30), a pure light yellow powder (0.563 g, 80.6%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:30. Figure 8 In the H NMR spectrum shown in the figure, the line corresponding to number 3 on the right is the random polymer P(N7-18 30 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0193]

[0194] Embodiment 32

[0195] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), monomer methacrylate n-octadecyl (0.2508 g, 740.7 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500 MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7-18 40 ), a pure light yellow powder (0.615 g, 80.8%), the structure of which is shown below, wherein the relationship between the degree of polymerization x and the degree of polymerization y is 70:40. Figure 8 In the H NMR spectrum shown in the figure, the line corresponding to number 4 on the right is the random polymer P(N7-18 40 ) H NMR spectrum; the peaks numbered 1, 2 and 3 in the structure shown are the hydrogen area peaks corresponding to the hydrophobic polymerization units, and the peaks numbered a to j correspond to the hydrogen areas of the molecular chains and other polymerization units.

[0196]

[0197] Comparative Example 1

[0198] Weigh poly(ethylene glycol) monomethyl ether (4-cyano-4-pentanoic acid dodecyl trithiocarbonate) (0.100 g, 18.5 μmol), monomer 2-(azepan-1-yl) ethyl methacrylate (0.410 g, 1943.1 μmol), initiator AIBN (1.35 mg, 8.23 ​​μmol) into a single-necked flask with a volume of 5 mL, and add 2 mL of N,N-dimethylacetamide. Seal the single-necked flask with a soft rubber stopper, then insert two needles, one end of which is below the liquid surface and the other end is above the liquid surface, blow argon gas below the liquid surface for 30 minutes, and then seal the bottle mouth with a sealing film. The flask was placed in an oil bath at 70°C for 24 hours. After the reaction, the flask was directly placed in a refrigerator at -20°C to quench the reaction. The reaction solution was then directly transferred to a dialysis bag (3500MWCO) and dialyzed with deionized water for 72 hours. After the dialysis, the product was freeze-dried to obtain a random polymer P(N7), a pure light yellow powder (0.432 g, 84.8%).

[0199] Embodiment 33

[0200] Weigh 10 mg of the random polymer P(N7-methyl) prepared in the above examples and comparative examples respectively. 10 )~P(N7-Eighteen 40 ) and P(N7) were added to a clean glass vial, and 4 mL of 0.2 M sodium dihydrogen phosphate solution was added to dissolve the mixture to prepare a 2.5 mg / mL solution. 40 μL of the solution was added with 160 μL of 0.2 M sodium dihydrogen phosphate under ultrasonic conditions to adjust the pH value to around 7.4 to form nanoparticles. The average particle size of the nanoparticles formed by each random polymer is as follows: Fig. 9 shown.

[0201] Embodiment 34

[0202] Weigh equal amounts of the random polymers described in Examples 1 to 32 and Comparative Example 1 and add them to a clean glass vial. Taking Comparative Example 1 as an example, 10 mg of the random polymer was dissolved in 4 mL of 0.2 M sodium dihydrogen phosphate solution to prepare a 2.5 mg / mL random polymer solution. Then take 400 μL of the random polymer solution, add 20 μL of an aqueous solution of the antigen OVA (10 mg / mL), and then add 1600 μL of 0.2 M sodium dihydrogen phosphate under ultrasonic conditions to make the pH value reach about 7.4 to obtain a random polymer P(N7-methyl)-1,2-dihydrogen phosphate solution. 10 )~P(N7-Eighteen 40) and P(N7) nanovaccines. Take 400μL of 0.2M sodium dihydrogen phosphate solution, add 20μL of antigen OVA aqueous solution (10mg / mL), and then add 1600μL of 0.2M disodium hydrogen phosphate under ultrasonic conditions to obtain the OVA group nanovaccine; take 400μL of 0.2M sodium dihydrogen phosphate solution, add 1600μL of 0.2M sodium dihydrogen phosphate solution under ultrasonic conditions, and record it as the Control group.

[0203] On day 0, mice were subcutaneously injected with 200 μL of the random polymer nanovaccine, which was labeled as P(N7- 10 )~P(N7-Eighteen 40 ) and P(N7) groups; the OVA group was injected with nanovaccine, recorded as the OVA group; the Control group was injected with solution, recorded as the Control group. On the second day, the inguinal lymph nodes of the mice were dissected and the mass of the lymph nodes was weighed. After mechanical disruption at 37°C, the samples were passed through a 200-mesh nylon mesh filter to obtain a single-cell suspension, and the cells were counted using a cell counting plate. Then they were incubated with anti-CD11c, anti-CD86 and anti-SIINFKEL-H2Kb antibodies at a concentration of 1:100 for 30 minutes at 4°C to analyze the antigen cross-presentation and activation effects of DCs in the lymph nodes. The samples acquired flow data on a flow cytometer (BD FACSCelesta) and were analyzed using FlowJo software. The results are shown in Figure 2. Fig.10 shown.

[0204] On day 0 and day 14, mice in the second group were subcutaneously injected with random polymers P(N7), P(N7-hexane 20 )~P(N7-H 40 )、P(N7-benzyl 10 )~P(N7-benzyl 30 ), P(N7- 10 ) and P(N7- 20 ) nanovaccine, OVA nanovaccine and control solution, the mice were dissected and spleens were removed on the 21st day. The samples were mechanically crushed at 37°C, and then filtered through a 200-mesh nylon filter to obtain a single-cell suspension, and the cells were counted using a cell counting plate. 2×10 6The cells were plated into a 48-well plate, and then the SIINFEKL polypeptide was added to the experimental system at a final concentration of 10 μg / mL. After 1 hour, the protein transport inhibitor Brefeldin A was added to reach a final concentration of 10 μg / mL. After 5 hours, the spleen cells were incubated with the dead and live dye Aqua, anti-CD3, and anti-CD8 for 30 minutes, and then the cells were stained with anti-IFN-γ for 30 minutes according to the intracellular factor staining method to analyze the antigen-specific immune response. The samples were used to obtain flow data on a flow cytometer (BD FACSCelesta) and analyzed by FlowJo software. The results are shown in Fig.11 shown.

[0205] Fig.10 middle, Fig.10 (a) The mass of draining lymph nodes after mice were injected with different nanovaccines. Fig.10 (b) Antigen cross-presentation ratio of DC cells in draining lymph nodes after mice were injected with different nanovaccines. Fig.10 (c) The activation ratio of DC cells in the draining lymph nodes of mice after injection of different nanovaccines. Fig.10 (a) It can be seen that mice injected with random polymer P(N7-methyl 10 )~P(N7-Eighteen 40 ) and P(N7) nanovaccines, the weight of the draining lymph nodes of mice more than doubled, from 2.7 mg and 2.6 mg in the Control group and OVA group to 5.4 mg-7.8 mg.

[0206] like Fig.10 As shown in (b), mice were injected with random polymer P(N7- 10 )~P(N7-Eighteen 40 ) and P(N7) nanovaccines, the antigen cross-presentation ratio increased significantly compared with the Control group and OVA group, and the injection of random polymer P(N7- 10 )~P(N7-Eighteen 40 ) nano-vaccine, the antigen cross-presentation ratio showed a clear structure-activity relationship. 10 )~P(N7-A 40 ) nanovaccine, the hydrophobic group is methyl, and as the proportion of methyl-containing polymer units increases, the antigen cross-presentation ratio gradually increases. 10 )~P(N7-H 40 ) nanovaccine, the hydrophobic group is hexyl. As the proportion of hexyl-containing polymer units increases, the antigen cross-presentation ratio increases first and then decreases. When the degree of polymerization is 20, the antigen cross-presentation ratio is the largest. 10 )~P(N7-Eighteen40 ) in the nanovaccine, the hydrophobic group is octadecyl. As the proportion of octadecyl-containing polymer units increases, the antigen cross-presentation ratio gradually decreases.

[0207] like Fig.10 (c) shows that mice were injected with random polymer P(N7-methyl 10 )~P(N7-Eighteen 40 ) and P(N7) nanovaccines, there was no significant difference in the activation ratio of DC cells, but it was significantly increased compared with the Control group and OVA group. 10 )~P(N7-A 40 ) nano-vaccine, the hydrophobic group is methyl, and as the proportion of methyl-containing polymer units increases, the activation ratio of DC cells tends to increase. 10 )~P(N7-H 40 ) in the nanovaccine, the hydrophobic group is hexyl. As the proportion of hexyl-containing polymer units increases, there is no significant difference in the activation ratio of DC cells.

[0208] In the random polymer P(N7-18 10 )~P(N7-Eighteen 40 ) in the nanovaccine, the hydrophobic group is octadecyl, and the hydrophobic group with a longer carbon chain will affect the activation function of the N7 unit. Therefore, as the proportion of octadecyl-containing polymer units increases, the activation ratio of DC cells tends to decrease.

[0209] Fig.11 (a) shows the proportion of antigen-specific T cells in mice after injection of different nanovaccines. Fig.11 (b) is the corresponding cell heat map. Compared with the Control group and the OVA group, the proportion of antigen-specific T cells increased significantly after the injection of the random polymer nanovaccine described in the embodiment and the comparative example. Compared with the P(N7) group, the proportion of antigen-specific T cells increased significantly after adding nanovaccines with different types and proportions of hydrophobic units, and showed a structure-activity relationship similar to that of the antigen cross-presentation experiment. When the hydrophobic group is hexyl and the polymerization ratio of the hydrophobic group-containing polymer unit is 20, the proportion of antigen-specific T cells is the highest (PN7-hexyl). 20 When the polymerization ratio of the polymerized units containing hexyl hydrophobic groups increases to 30 and 40 (PN7-hexyl 30 and PN7-hexadecene 40 ), the proportion of antigen-specific T cells decreased. When the hydrophobic group is octyl, the polymerization ratio of the hydrophobic group-containing polymerization unit increases from 10 to 20, and the proportion of antigen-specific T cells increases (PN7-octyl). 10 and PN7-xin 20 ).

[0210] In summary, the random polymer modified with hydrophobic groups provided by the present invention has a significant structure-activity relationship between the type and quantity of hydrophobic groups and the DC cell activation, antigen cross-presentation ratio and antigen-specific T cell ratio induced by nanovaccines. By increasing the level of antigen cross-presentation, the level of antigen-specific immune response can be further improved and the immune efficacy can be enhanced.

Claims

1. A random polymer comprising a hydrophilic structure represented by formula (I), a first repeating unit represented by formula (II) and a second repeating unit represented by formula (III), in, n is an integer selected from 2 to 500; m is an integer selected from 1 to 20; the molar ratio of the first repeating unit to the second repeating unit is (1 to 150):(1 to 150); R5 is selected from C1~C 30 Alkyl, C1~C 30 Alkyl, C2~C 30 Alkenyl, C2-C 30 Alkenyl, C2~C 30 Alkynyl, C2-C 30 Alkynyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C3~C 30 cycloalkenyl, C3~C 30 Cycloalkenyl, C2~C 30 heterocyclic alkenyl, C2-C 30 Heterocycloalkenyl, C3~C 30 cycloalkynyl, C3~C 30 Cycloalkynyl, C2~C 30 heterocycloalkynyl, C2-C 30 Heterocycloalkynyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 Alkenyl, C2~C 30 Alkynyl, C2~C 30 At least one of an amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl group; R2 and R3 are independently selected from hydrogen, C1-C 30 Alkyl, C1~C 30 Alkyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 at least one of amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl; or R2 and R3 and the N atom to which they are connected together form a 3-30 membered heterocyclic ring; R1 and R4 are independently selected from hydrogen, C1-C 30 Alkyl, C1~C 30 Alkyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 At least one of amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl groups.

2. The random polymer according to claim 1, characterized in that The R5 is selected from C1~C 25 Alkyl, C1~C 25 Alkyl, C2~C 25 Alkenyl, C2-C 25 Alkenyl, C2~C 25 Alkynyl, C2-C 25 Alkynyl, C3~C 25 cycloalkyl, C3~C 25 Cycloalkyl, C3~C 25 cycloalkenyl, C3~C 25 Cycloalkenyl, C3~C 25 cycloalkynyl, C3~C 25 Cycloalkynyl, C6~C 25 C6~C 25 The aromatic group, the substituent is selected from C1 to C 25 Alkyl, C2~C 25 Alkenyl, C2~C 25 Alkynyl and C6~C 25 At least one of the aromatic groups; The R2 and R3 are independently selected from C1-C5 alkyl groups or C1-C5 alkyl groups substituted with at least one substituent, wherein the substituent is selected from at least one of amino, hydroxyl, carboxyl, cyano and thiol; or R2, R3 and the N atom where they are located together form a ring having the following structure: wherein a, b, c, d and e are independently selected from integers of 1 to 5; R a and R b Independently selected from hydrogen or C1-C5 alkyl; —X—is selected from —O—, R c Selected from hydrogen or C1-C5 alkyl; The R1 and R4 are independently selected from hydrogen, C1-C 25 Alkyl, C3~C 25 Cycloalkyl, C6~C 25 Aromatic or C6~C 25 The heteroaryl group.

3. The random polymer according to claim 2, characterized in that The R5 is selected from C1~C 20 alkyl, C1~C 20 Alkyl, C2~C 20 or at least one substituent substituted with C2-C 20 The substituent is selected from C1 to C 20 Alkyl, C2~C 20 Alkenyl, C3~C 20 Cycloalkyl or C6~C 20 Aromatic groups; The R2, R3 and the N atom to which they are connected together form a ring having the following structure: Wherein, a is selected from an integer of 1 to 5; R1 and R4 are independently selected from C1 to C 20 of alkyl.

4. The random polymer according to claim 3, characterized in that The R5 is selected from the following structures: Wherein, f is selected from an integer of 0 to 20; R d Selected from hydrogen, C2~C 20 Alkenyl, C3~C 20 Cycloalkyl or C6~C 20 Aromatic base.

5. The random polymer according to claim 1, characterized in that It has the following structure: wherein x and y are independently selected from integers of 2 to 300; f is selected from integers of 0 to 20; R d is selected from hydrogen or phenyl.

6. A method for preparing a random polymer, characterized in that: include: A macromolecular chain transfer agent having a structure of formula (IV), a monomer having a structure of formula (V), a monomer having a structure of formula (VI) and an initiator undergo a polymerization reaction to obtain the random polymer; wherein n is selected from an integer of 2 to 500; m is selected from an integer of 1 to 20; R5 is selected from C1~C 30 alkyl, C1~C 30 Alkyl, C2~C 30 Alkenyl, C2-C 30 Alkenyl, C2~C 30 Alkynyl, C2-C 30 Alkynyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C3~C 30 cycloalkenyl, C3~C 30 Cycloalkenyl, C2~C 30 heterocyclic alkenyl, C2-C 30 Heterocycloalkenyl, C3~C 30 cycloalkynyl, C3~C 30 Cycloalkynyl, C2~C 30 heterocycloalkynyl, C2-C 30 Heterocycloalkynyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 Alkenyl, C2~C 30 Alkynyl, C2~C 30 At least one of an amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl group; R2 and R3 are independently selected from hydrogen, C1-C 30 alkyl, C1~C 30 Alkyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 at least one of amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl; or R2 and R3 and the N atom to which they are connected together form a 3-30 membered heterocyclic ring; R1, R4 and R6 are independently selected from hydrogen, C1-C 30 alkyl, C1~C 30 Alkyl, C3~C 30 cycloalkyl, C3~C 30 Cycloalkyl, C2~C 30 heterocycloalkyl, C2-C 30 Heterocycloalkyl, C6~C 30 aromatic group, C6~C 30 Aromatic groups, C6~C 30 heteroaryl, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, at least one substituent substituted C1~C 30 Alkoxy, C1~C 30 alkylthio, C1~C 30 Alkylthio, C2~C 30 ether, C2~C 30 Ether, C1~C 30 esters, C1~C 30 Esters, C2~C 30 sulfide, C2~C 30 Sulfide, C2~C 30 amine or at least one substituent substituted C2~C 30 The amine, the substituent is selected from C1 to C 30 Alkyl, C6~C 30 Aromatic groups, C6~C 30 Heteroaryl, C1~C 30 Alkoxy, C1~C 30 Alkylthio, C2~C 30 Ether, C1~C 30 Esters, C2~C 30 Sulfide, C2~C 30 At least one of amine, hydroxyl, carboxyl, aldehyde, carbonyl, amino, cyano, thiol and thiocarbonyl groups.

7. The preparation method according to claim 6, characterized in that: The solvent for the polymerization reaction is selected from one or more of dioxane, N,N-dimethylacetamide and N,N-dimethylformamide, and the mass ratio of the solvent to the macromolecular chain transfer agent is 10 to 30:

1.

8. The preparation method according to claim 6, characterized in that: The initiator is selected from azobisisobutyronitrile or azobiscyanovaleric acid, and the molar ratio of the initiator to the macromolecular chain transfer agent is 1:2-5.

9. The preparation method according to claim 6, characterized in that: The polymerization reaction is carried out under anaerobic conditions; The polymerization reaction temperature is 35-70° C. and the reaction time is 12-48 hours.

10. A nanoparticle, characterized in that: The random polymer is formed by the random polymer described in any one of claims 1 to 5 or the random polymer prepared by the preparation method of any one of claims 6 to 9.

11. A nanoadjuvant, characterized in that: The invention comprises the random polymer described in any one of claims 1 to 5, the random polymer prepared by the preparation method of any one of claims 6 to 9, or the nanoparticles described in claim 10.

12. A nano vaccine, characterized in that: The invention comprises the nano adjuvant according to claim 11, an antigen and an adjuvant acceptable to the vaccine.

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