CN111378146A - Polymer, nanogel for carrying protein drug and application of nanogel - Google Patents
Polymer, nanogel for carrying protein drug and application of nanogel Download PDFInfo
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- CN111378146A CN111378146A CN202010053145.3A CN202010053145A CN111378146A CN 111378146 A CN111378146 A CN 111378146A CN 202010053145 A CN202010053145 A CN 202010053145A CN 111378146 A CN111378146 A CN 111378146A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
- C08G83/008—Supramolecular polymers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
- A61K47/6951—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Abstract
The invention provides a polymer which has a structure shown in a formula (I). The polymer designed by the invention not only has the characteristics ofA fixed structure and can have a group of compounds having a hydrophobic inner cavity or a group of compounds which can be inserted into a hydrophobic inner cavity. The polymers respectively containing the two groups form a network-shaped nano polymer composite material through the supermolecule crosslinking effect, and when the network-shaped nano polymer composite material is used as a nano gel carrier for loading protein drugs, the protein drugs are encapsulated in a crosslinking grid while the crosslinking network is formed through the supermolecule crosslinking effect, so that the protein drugs effectively play a role in vivo. The nanogel does not modify proteins, does not influence the activity of the proteins, can be realized in a mild aqueous solution, effectively protects protein drugs, maintains the stability of the protein drugs and improves the in vivo circulation time of the protein drugs.
Description
Technical Field
The invention belongs to the technical field of protein drug loading, and relates to a polymer, a polymer material, a protein drug-loaded composite material and application, in particular to a polymer, nanogel for loading a protein drug and application.
Background
Since recombinant insulin was the first FDA-approved recombinant protein, protein drugs have been used in disease therapy, such as endostatin, cytokines, monoclonal antibodies, cytochrome C, etc., which are widely used in cancer, metabolic disorders, autoimmune diseases, etc. The wide application of protein drugs is mainly due to the advantages of good biological activity, high specificity, low toxicity and the like of the protein drugs. Despite the growth of protein drugs, there are still many challenges to the use of protein drugs. The protein medicine has poor stability and is easy to deteriorate under the influence of temperature, pH, organic solvent and the like. The circulation time of the protein drug in the body is short, and the cell membrane penetrability of the protein drug is poor. Therefore, the carrier is used for coating the protein medicine, and the improvement of the bioavailability of the protein medicine is of great significance.
The main strategy currently adopted by researchers to solve the problems of the drawbacks of protein drugs is polyethylene glycol (PEG) conversion of protein drugs. Gaertner et al oxidized the end of interleukin 8(IL-8) with sodium periodate to form an aldehyde group, then reacted with the end hydroxylamino group of PEG to form an oxime bond, to construct PEGylated IL-8, to prolong the in vivo circulation time of IL-8, to enhance the enrichment of IL-8 in tumor sites, to reduce the toxic and side effects of IL-8, and to greatly improve the in vivo utilization of IL-8 [ Gaertner, H.F.; offord, R.E.bioconjugate chem.1996,7,38 ]. Abuchowski et al, by using a method of covalently bonding bovine serum albumin with PEG, greatly increase the circulation time of bovine serum albumin in vivo, reduce the immunogenicity in vivo, improve the in vivo stability, and reduce the renal clearance rate, thereby prolonging the half-life of the protein drug [ Abuchowski, a.; vanes, t.; palczuk, n.c.; davis, f.f.j.biol.chem.1977,252,3578 ], but the method has serious disadvantages that firstly, single-point modified pegylated protein is difficult to separate from unmodified and multi-point modified protein, and simultaneously, some complex chemical reactions are introduced to cause unpredictable safety problems, and most importantly, the protein modification method is easy to cause the change of the spatial structure of the protein so as to reduce the activity of the protein medicament.
Therefore, how to find a suitable way and material to carry protein drugs, which solves the above-mentioned drawbacks of the existing carrier-coated protein drugs, has become one of the focuses of research of many leading-edge scholars in the technical field of tumor detection.
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide a polymer, a polymer material, a protein drug-loaded composite material, and an application thereof, and particularly to a polymer material for loading a protein drug, wherein the polymer material provided by the present invention is prepared into a nanogel as a drug carrier, which does not undergo any modification on a protein, does not affect the activity of the protein, can be realized in a mild aqueous solution, can effectively protect the protein drug, maintain the stability thereof, improve the in vivo circulation time of the protein drug, and is beneficial to the targeted enrichment of the protein drug at a tumor site and greatly improve the endocytosis efficiency of cells to the protein drug. And the non-covalent protein loading can be realized under mild conditions, the method is simple, the conditions are controllable, and the method is suitable for large-scale popularization and application.
The invention provides a polymer, which has a structure shown in a formula (I);
wherein R includes R1Or R2；
The R is1Selected from compounds having a hydrophobic internal cavity;
the R is2Selected from compounds that can intercalate into hydrophobic lumens;
L1、L2and L3Each independently selected from-CH2-or-CH2CH2-；
x, y and z are integers, x is more than or equal to 0, y is more than 0, z is more than 0, and x + y + z is more than or equal to 10 and less than or equal to 5000;
n is an integer, and n is more than or equal to 10 and less than or equal to 500.
Preferably, the compound having a hydrophobic inner cavity comprises one or more of cyclodextrin, calixarene and cucurbituril;
the compound capable of being embedded into the hydrophobic inner cavity comprises one or more of azobenzene, adamantane and biphenyl;
x + y + z is more than or equal to 30 and less than or equal to 300;
the R is1And R2Has the function of realizing the cross-linking of supermolecule effect.
The invention provides a polymer material, which is obtained by crosslinking a polymer with a structure shown in a formula (II) and a polymer with a structure shown in a formula (III);
wherein R is1Selected from compounds having a hydrophobic internal cavity;
R2selected from compounds that can intercalate into hydrophobic lumens;
L1、L2and L3Each independently selected from-CH2-or-CH2CH2-；
x, y and z are integers, x is more than or equal to 0, y is more than 0, z is more than 0, and x + y + z is more than or equal to 10 and less than or equal to 5000;
n is an integer, and n is more than or equal to 10 and less than or equal to 500.
Preferably, the crosslinking is R in a polymer having a structure represented by formula (II)1And a polymer having a structure represented by the formula (III)R in (1)2Cross-linking by supramolecular interaction;
the compound with a hydrophobic inner cavity comprises one or more of cyclodextrin, calixarene and cucurbituril;
the compound which can be embedded into the hydrophobic inner cavity comprises one or more of azobenzene, adamantane and biphenyl.
Preferably, after the crosslinking, the compound with a hydrophobic inner cavity in the polymer with the structure shown in the formula (II) and the compound capable of being embedded into the hydrophobic inner cavity in the polymer with the structure shown in the formula (III) are physically crosslinked to form a supramolecular network structure through host-guest interaction;
x + y + z is more than or equal to 30 and less than or equal to 300;
the polymer material has the function of carrying the drug.
The invention provides a composite material for carrying protein drugs, which comprises the polymer material and the protein drugs in any one of the technical schemes.
Preferably, the protein drug comprises one or more of cytokines, chemokines, monoclonal antibodies and fragments thereof, and therapeutic protein drugs;
the molar ratio of the protein drug to the polymer material is 1: (0.05-20);
the protein drug is encapsulated in a cross-linked network of cross-linked meshes formed by the polymer material;
the composite material includes a nanomaterial.
Preferably, the size of the composite material is 50-1000 nm;
the composite material comprises a nanogel;
the protein drug is encapsulated inside the nanogel;
the protein medicine comprises one or more of RNase, interleukin 2, interleukin 12 and cytochrome C;
the protein drug-loaded composite material is obtained by mixing and reacting a polymer solution with a structure shown in a formula (II), a polymer solution with a structure shown in a formula (III) and a protein drug.
Preferably, the solution comprises a buffer solution;
the mixing reaction time is 2-48 h;
the molar ratio of the polymer having the structure represented by the formula (II) to the polymer having the structure represented by the formula (III) is 1: (0.1 to 20);
the molar ratio of the protein drug to the polymer having the structure represented by formula (II) is 1: (0.1 to 20);
the concentration of the polymer solution with the structure shown in the formula (II) is 0.01-10 mM;
the concentration of the polymer solution with the structure shown in the formula (III) is 0.01-10 mM.
The invention provides the application of the polymer in any one of the technical schemes, the polymer material in any one of the technical schemes or the protein-carrying medicine composite material in any one of the technical schemes in the fields of tumor treatment, metabolic disorder and autoimmune system diseases.
The invention provides a polymer, which has a structure shown in a formula (I); wherein R includes R1Or R2(ii) a The R is1Selected from compounds having a hydrophobic internal cavity; the R is2Selected from compounds that can intercalate into hydrophobic lumens; l is1、L2And L3Each independently selected from-CH2-or-CH2CH2-; x, y and z are integers, x is not less than 0 and y>0，z>0, 10 is less than or equal to x + y + z is less than or equal to 5000; n is an integer, and n is more than or equal to 10 and less than or equal to 500. Compared with the prior art, the invention aims at the problems of difficult estimated safety of carrier-coated protein drugs in the application field of the existing protein drugs and the reduction of the activity of the protein drugs caused by the change of the spatial structure of the protein.
The invention selects the nano-gel as the direction of improvement and application among various carrier materials based on the advantages of the nano-gel, wherein the nano-gel is an intramolecular cross-linked polymer gel existing in the form of nano-particles (the particle diameter is 1-1000 nm), the internal structure of the nano-gel is a typical grid structure, and the nano-gel can be dispersed into nano-sized hydrogel particles in an aqueous solution. Like conventional polymer hydrogels, such soft materials of nanometer dimensions can undergo many physicochemical changes such as porosity, rheology, refractive index, surface charge density and colloidal stability according to the external environment such as temperature, pH, solvent, applied stress, light intensity, electromagnetic field or various chemical and ionic strength stimuli. The nano gel has wide application prospect in the aspects of drug controlled release, biomedical engineering, diagnostic analysis, enzyme immobilization, enzyme activity regulation and control and the like due to the intelligent response performance, and is particularly suitable for encapsulating protein drugs due to the characteristics of high water content, high biocompatibility and large specific surface area. For example, Deng et al, constructed nanogel by ultraviolet light-induced click reaction using hyaluronic acid grafted cysteamine methacrylate and hyaluronic acid grafted lysine tetrazole, carried cytotoxic drugs telomerase B and cytochrome C. The nanogel effectively increases the endocytosis of the protein drug by the cells, and the nano telomerase B has longer circulation time in vivo and better enrichment at a tumor part. However, there is a corresponding problem in that the method of constructing nanogel by ultraviolet light easily induces inactivation and denaturation of proteins.
The invention is based on the finding that polymers having the structure of formula (I) are designed, which not only have a specific structure, but also are capable of having groups of compounds having a hydrophobic inner cavity or groups of compounds which can be inserted into a hydrophobic inner cavity. The polymer (II) and the polymer (III) respectively containing the groups can form a network-shaped nano polymer composite material through supermolecular crosslinking. Furthermore, the polymer composite material can be used as a nano gel carrier for loading protein drugs, and when nano gel is crosslinked through the supermolecule effect, the protein drugs are encapsulated in a crosslinked grid while a crosslinked network is formed, so that the protein drugs effectively play a role in vivo. Effectively solves the problems that protein drugs are easy to deteriorate and inactivate, have short in-vivo circulation time and are difficult to be endocytosed by cells, and has wide development prospect in the aspect of realizing in-vivo application of the protein drugs. The nanogel provided by the invention is used as a drug carrier, is not modified on protein, does not influence the activity of the protein, can be realized in a mild aqueous solution, can effectively protect the protein drug, maintains the stability of the protein drug, improves the in vivo circulation time of the protein drug, is beneficial to the targeted enrichment of the protein drug at a tumor part, and greatly improves the endocytosis efficiency of cells to the protein drug.
The polymer material provided by the invention can be used for preparing nanogel, a cross-linked grid is formed through the interaction of a host and an object, and a protein drug is encapsulated in the nanogel, so that the protein drug can effectively circulate in vivo for a long time, is enriched at a tumor part and slowly releases the protein drug, and the capability of the protein drug taken up by cells is greatly enhanced. Most importantly, in the process of loading the protein drug by the nanogel, the protein drug is carried out under the conditions of aqueous solution, proper temperature, pH and the like, and the biological activity of the protein drug is efficiently maintained. The method of loading the protein drug by the nanogel effectively solves the problems of short circulation time, poor endocytosis and easy inactivation and deterioration of the protein drug in vivo, and has wide development prospect in the field of tumor treatment. Moreover, the preparation method provided by the invention is simple, the raw material source is wide, the batch production can be realized, and the industrialization can be realized.
The experimental result shows that the polyglutamic acid grafted polyethylene glycol grafted aminoazobenzene prepared by the invention and the polyglutamic acid grafted polyethylene glycol grafted amino β cyclodextrin carry the RNA enzyme, the size of the obtained RNA enzyme loaded nanogel is between 100 and 200nm, on a cell level, the endocytosis of the RNA enzyme loaded nanogel or pure RNA enzyme by the cell is compared, and the result shows that the endocytosis efficiency of the RNA enzyme is improved by more than two times by the nanogel.
Drawings
FIG. 1 is a diagram showing the preparation of polyethylene glycol-grafted poly-L-glutamic acid obtained in example 31A HNMR map;
FIG. 2 is the NMR spectrum of polyglutamic acid grafted PEG-grafted azobenzene prepared in example 4;
FIG. 3 is the NMR chart of the polyglutamic acid grafted PEG-grafted aminocyclodextrin prepared in example 5;
FIG. 4 shows the results of dynamic light scattering in water at a concentration of 0.2mg/mL in the RNase-supported nanogel prepared in example 6 of the invention;
FIG. 5 is a graph comparing the flow results measured in example 8 of the present invention;
FIG. 6 is a pharmacokinetic profile measured in example 9 of the present invention.
Detailed Description
For a further understanding of the invention, preferred embodiments of the invention are described below in conjunction with the examples, but it should be understood that these descriptions are included merely to further illustrate the features and advantages of the invention and are not intended to limit the invention to the claims.
All of the starting materials of the present invention, without particular limitation as to their source, may be purchased commercially or prepared according to conventional methods well known to those skilled in the art.
All the raw materials of the present invention are not particularly limited in purity, and the present invention preferably adopts a purity which is conventional in analytical purification, the medical field or the field of application of protein drug loading thereof.
All the noun expressions and acronyms of the invention belong to the conventional noun expressions and acronyms in the field, each noun expression and acronym is clearly and definitely clear in the relevant application field, and a person skilled in the art can clearly, exactly and uniquely understand the noun expressions and acronyms.
The invention provides a polymer, which has a structure shown in a formula (I);
wherein R includes R1Or R2；
The R is1Selected from compounds having a hydrophobic internal cavity;
the R is2Is selected fromCompounds that can intercalate into hydrophobic lumens;
L1、L2and L3Each independently selected from-CH2-or-CH2CH2-；
x, y and z are integers, x is more than or equal to 0, y is more than 0, z is more than 0, and x + y + z is more than or equal to 10 and less than or equal to 5000;
n is an integer, and n is more than or equal to 10 and less than or equal to 500.
In the present invention, the substituent R includes R1Or R2. Preferably, said R is1And R2Has the function of realizing the cross-linking of supermolecule effect.
The invention in principle applies to said substituents R1The specific choice of the compound with a hydrophobic inner cavity is not particularly limited, and can be adjusted by a person skilled in the art according to application conditions, application requirements or product performance requirements, the invention is used for improving the capability of the polymer as a protein drug carrier, not modifying protein and not influencing protein activity, and can more effectively protect the protein drug, maintain the stability of the protein drug, and further improve the endocytosis efficiency of cells to the protein drug, and the compound with the hydrophobic inner cavity preferably comprises one or more of cyclodextrin, calixarene and cucurbituril, more preferably cyclodextrin, calixarene or cucurbituril, more preferably cyclodextrin or calixarene of C3-C8, and more preferably cyclodextrin.
The invention in principle applies to said substituents R2The specific choice of the compound capable of being embedded into the hydrophobic cavity is not particularly limited, and can be adjusted by a person skilled in the art according to application conditions, application requirements or product performance requirements.
In the present invention, said L1、L2And L3Each independently selected from-CH2-or-CH2CH2-, i.e. L1Is selected from-CH2-or-CH2CH2-。L2Is selected from-CH2-or-CH2CH2-。L3Is selected from-CH2-or-CH2CH2-。
In the present invention, x, y and z are polymerization degrees and are selected from integers, and preferably, x, y and z are in a random form. Wherein x is more than or equal to 0, y is more than 0, z is more than 0, 10 is more than or equal to x + y + z is less than or equal to 5000, more preferably 15 is more than or equal to x + y + z is less than or equal to 3000, more preferably 20 is more than or equal to x + y + z is less than or equal to 1000, more preferably 25 is more than or equal to x + y + z is less than or equal to 500, and more preferably 30 is more than or equal to x + y + z is less than or equal. Specifically, x + y + z is more than or equal to 50 and less than or equal to 250, specifically, x + y + z is more than or equal to 75 and less than or equal to 200, and specifically, x + y + z is more than or equal to 100 and less than or equal to 150. Wherein y is greater than 0, z is greater than 0, x is greater than or equal to 0, preferably, x is greater than 10; y > 20; z > 4.
Preferably, n is 20. ltoreq. n.ltoreq.400, more preferably 50. ltoreq. n.ltoreq.260, and most preferably 80. ltoreq. n.ltoreq.180.
In the present invention, n is a polymerization degree and is selected from integers. Wherein 10. ltoreq. n.ltoreq.500, more preferably 20. ltoreq. n.ltoreq.400, more preferably 30. ltoreq. n.ltoreq.300, more preferably 50. ltoreq. n.ltoreq.260, most preferably 80. ltoreq. n.ltoreq.180.
The preparation process of the polymer having the structure shown in formula (I) is not particularly limited in the present invention, and the preparation method of the polymer having such a structure is well known to those skilled in the art, and those skilled in the art can adjust the preparation process according to the application situation, application requirements or product performance requirements, and the preparation process of the polymer having the structure shown in formula (I) is preferably performed by referring to the methods in the published documents of the corresponding structures or based on the preparation processes in the embodiments of the present invention.
The invention provides a polymer material, which is obtained by crosslinking a polymer with a structure shown in a formula (II) and a polymer with a structure shown in a formula (III);
wherein R is1Selected from compounds having a hydrophobic internal cavity;
R2selected from compounds that can intercalate into hydrophobic lumens;
L1、L2and L3Each independently selected from-CH2-or-CH2CH2-；
x, y and z are integers, x is more than or equal to 0, y is more than 0, z is more than 0, and x + y + z is more than or equal to 10 and less than or equal to 5000;
n is an integer, and n is more than or equal to 10 and less than or equal to 500.
The structure and group selection and parameters in the structural formula of the raw materials ((II) and (III)) of the polymer material of the invention and the corresponding preferred principles thereof preferably correspond to the structure and group selection and parameters in the structural formula of the polymer and the corresponding preferred principles thereof one by one, and are not described in detail herein.
The invention is not particularly limited in principle to the specific mode of the cross-linked compound, and can be adjusted by those skilled in the art according to the application condition, application requirement or product performance requirement, in order to improve the ability of the polymer as a protein drug carrier, not modifying the protein and not affecting the activity of the protein, and can more effectively protect the protein drug, maintain the stability thereof, and further improve the endocytosis efficiency of the protein drug by the cell, the cross-linking is preferably R in the polymer having the structure shown in formula (II)1And R in the polymer having a structure represented by the formula (III)2Cross-linking by supramolecular interaction.
Preferably, the polymer having the structure represented by the formula (II) may specifically be of the following structure:
preferably, the polymer having the structure represented by the formula (III) may specifically be of the following structure:
specifically, the azobenzene-cyclodextrin supramolecular polymer material is obtained after the two polymers are crosslinked.
The specific structure of the cross-linked compound is not particularly limited in principle, and can be adjusted by a person skilled in the art according to application conditions, application requirements or product performance requirements.
The specific ratio of the polymer having the structure shown in formula (II) and the polymer having the structure shown in formula (III) is not particularly limited in the present invention, and can be adjusted by those skilled in the art according to the application situation, application requirement or product performance requirement, and in order to improve the ability of the polymer as a protein drug carrier, not modify the protein and not affect the activity of the protein, and to protect the protein drug more effectively, maintain its stability, and further improve the endocytosis efficiency of the protein drug by the cell, the molar ratio of the polymer (II) and the polymer (III) is preferably 1: (0.1 to 20), more preferably 1: (0.2 to 15), more preferably 1: (0.3 to 10), more preferably 1: (0.4 to 5), more preferably 1: (0.5-2).
The preparation process of the polymer material is preferably referred to the preparation process of the protein-drug-loaded composite material, and no protein drug is added in the preparation process of the polymer material.
The objective state of the polymer material is not particularly limited in principle, and can be adjusted by a person skilled in the art according to application conditions, application requirements or product performance requirements. The polymer material of the present invention preferably has a drug-supporting function, and more preferably, a nanogel having a drug-supporting function.
The preparation process of the polymer having the structure shown in formula (II) or the polymer having the structure shown in formula (III) is not particularly limited, and the preparation method of the polymer having such a structure, which is well known to those skilled in the art, can be adjusted by those skilled in the art according to the application situation, application requirements or product performance requirements, and the preparation method of the polymer having the structure shown in formula (II) is preferably referred to in the published literature of the corresponding structure or is based on the preparation process in the embodiment of the present invention.
The polymer with the structure shown in the formula (I), the polymer (II) containing the group of the compound with the hydrophobic inner cavity and the polymer (III) containing the group of the compound capable of being embedded into the hydrophobic inner cavity are provided in the steps of the invention, the polymer (II) and the polymer (III) can form a network-shaped nano polymer composite material through supermolecule cross-linking action, and further can selectively obtain nano gel, and a cross-linked grid is formed through the interaction of a host and an object, so that the protein drug can be encapsulated in the nano gel subsequently, the protein drug can effectively circulate in vivo for a long time, is enriched at a tumor part and slowly releases the protein drug, and the capability of the protein drug taken up by cells is greatly enhanced. The polymer composite material provided by the invention is used as a drug carrier, is not modified on protein, does not influence the activity of the protein, can be realized in a mild aqueous solution, can effectively protect the protein drug, maintains the stability of the protein drug, improves the in vivo circulation time of the protein drug, is beneficial to the targeted enrichment of the protein drug at a tumor part, and greatly improves the endocytosis efficiency of cells to the protein drug.
The invention also provides a composite material for carrying the protein drug, which comprises the polymer material and the protein drug in any one of the technical schemes.
The structure, the selection and the parameters of the groups of the polymer material in the composite material of the present invention, and the corresponding preferred principles thereof preferably correspond to the structure, the selection and the parameters of the groups of the polymer material, and the corresponding preferred principles thereof one by one, and are not described in detail herein.
The specific choice of the protein drug (protein) is not particularly limited in principle, and can be adjusted by those skilled in the art according to the application, application requirements or product performance requirements, and the invention is to improve the ability of the polymer as a protein drug carrier, not to modify the protein and not to affect the activity of the protein, and to protect the protein drug more effectively, maintain the stability thereof, and further improve the endocytosis efficiency of the protein drug by the cells, wherein the protein drug, i.e. the protein, preferably comprises one or more of cytokines, chemokines, monoclonal antibodies and fragments thereof, and therapeutic protein drugs, more preferably comprises one or more of cytokines, chemokines, monoclonal antibodies and fragments thereof, or therapeutic protein drugs, and specifically comprises one or more of rnase, interleukin 2, interleukin 12 and cytochrome C, more preferably RNase, interleukin 2, interleukin 12, and cytochrome C, Interleukin 12 or cytochrome C, more preferably RNase or interleukin 2.
In order to improve the ability of the polymer as a protein drug carrier, not modifying the protein and not affecting the activity of the protein, and more effectively protecting the protein drug, maintaining the stability of the protein drug, and further improving the endocytosis efficiency of the protein drug by cells, the molar ratio of the protein drug to the polymer material is preferably 1: (0.05-20), more preferably 1: (0.1 to 18), more preferably 1: (0.2 to 15), more preferably 1: (0.3 to 10), more preferably 1: (0.4 to 5), more preferably 1: (0.5-2).
The invention has no special limitation on the loading state of the protein drug in principle, and a person skilled in the art can adjust the loading state according to application conditions, application requirements or product performance requirements.
The invention has no particular limitation on the specific form of the protein drug-loaded composite material in principle, and can be adjusted by a person skilled in the art according to the application condition, application requirements or product performance requirements. Further, the composite material preferably comprises a nanogel, namely a nanogel carrying a protein drug, and particularly, the protein drug is preferably encapsulated in the nanogel.
The specific parameters of the protein drug-loaded composite material are not particularly limited in principle, and can be adjusted by a person skilled in the art according to application conditions, application requirements or product performance requirements, in order to improve the capacity of a polymer used as a protein drug carrier, not modifying protein and not influencing protein activity, and can more effectively protect a protein drug, maintain the stability of the protein drug, and further improve the endocytosis efficiency of cells to the protein drug, the size of the composite material is preferably 50-1000 nm, more preferably 100-800 nm, more preferably 200-700 nm, more preferably 300-600 nm, and more preferably 400-500 nm.
The preparation process of the protein drug-loaded composite material is not particularly limited in principle, and can be adjusted by a person skilled in the art according to application conditions, application requirements or product performance requirements.
The ratio of the polymer having the structure shown in formula (II) to the polymer having the structure shown in formula (III) is not particularly limited in the present invention, and can be adjusted by those skilled in the art according to the application, application requirements or product performance requirements, and in the present invention, in order to improve the ability of the polymer as a protein drug carrier, not modify the protein and not affect the activity of the protein, and to protect the protein drug more effectively, maintain its stability, and further improve the endocytosis efficiency of the protein drug by the cell, the molar ratio of the polymer having the structure shown in formula (II) to the polymer having the structure shown in formula (III) is preferably 1: (0.1 to 20), more preferably 1: (0.2 to 15), more preferably 1: (0.3 to 10), more preferably 1: (0.4 to 5), more preferably 1: (0.5-2).
In order to improve the ability of the polymer as a protein drug carrier, not modifying the protein and not affecting the activity of the protein, and more effectively protecting the protein drug, maintaining the stability thereof, and further improving the endocytosis efficiency of the protein drug by the cell, the molar ratio of the protein drug to the polymer having the structure shown in formula (II) is preferably 1: (0.1 to 20), more preferably 1: (0.2 to 15), more preferably 1: (0.3 to 10), more preferably 1: (0.4 to 5), more preferably 1: (0.5-2).
The solution selection and parameters of the polymer solution having the structure shown in formula (II) are not particularly limited in the present invention, and may be adjusted by those skilled in the art according to the application situation, application requirement or product performance requirement. Such as PB phosphate buffer. The concentration of the polymer solution having the structure represented by the formula (II) is preferably 0.01 to 10mM, more preferably 0.02 to 5mM, more preferably 0.02 to 1mM, more preferably 0.03 to 0.5mM, and more preferably 0.05 to 0.15 mM.
The solution selection and parameters of the polymer solution having the structure shown in formula (III) are not particularly limited in the present invention, and may be adjusted by those skilled in the art according to the application situation, application requirement or product performance requirement. Such as PB phosphate buffer. The concentration of the polymer solution having the structure represented by the formula (III) is preferably 0.01 to 10mM, more preferably 0.02 to 5mM, more preferably 0.02 to 1mM, more preferably 0.03 to 0.5mM, and more preferably 0.05 to 0.15 mM.
The specific parameters of the mixing reaction (crosslinking reaction) are not particularly limited in principle, and can be adjusted by a person skilled in the art according to application conditions, application requirements or product performance requirements, in order to improve the ability of the polymer as a protein drug carrier, not modify protein and not influence the activity of the protein, and can more effectively protect the protein drug, maintain the stability of the protein drug, and further improve the endocytosis efficiency of the protein drug by cells, the time of the mixing reaction is preferably 2-48 h, more preferably 8-42 h, more preferably 14-36 h, and more preferably 20-30 h. The temperature of the mixing reaction is preferably normal temperature, and specifically can be 5-40 ℃, or 10-35 ℃, or 15-30 ℃, or 20-25 ℃.
The invention also provides the application of the polymer in any one of the technical schemes, the polymer material in any one of the technical schemes or the protein-carrying medicine composite material in any one of the technical schemes in the fields of tumor treatment, metabolic disorder and autoimmune system diseases.
The steps of the invention provide a polymer, nanogel for carrying protein drugs and application.
The present invention provides a polymer having a structure represented by formula (I), which has not only a specific structure but also a group of compounds capable of having a hydrophobic inner cavity or a group of compounds capable of being intercalated into a hydrophobic inner cavity. The polymer (II) and the polymer (III) respectively containing the groups can form a network-shaped nano polymer composite material through supermolecular crosslinking. Furthermore, the polymer composite material is prepared into a nano gel carrier for loading protein drugs, the protein drugs are added into the nano gel carrier during preparation, and the protein drugs are encapsulated in a cross-linked grid through supermolecule effect, so that the obtained nano protein drugs can effectively circulate in vivo for a long time, are enriched at tumor sites and slowly release the protein drugs, and the capability of the protein drugs to be taken by cells is greatly enhanced. Most importantly, in the process of loading the protein drug by the nanogel, the protein drug is carried out under the conditions of aqueous solution, proper temperature, pH and the like, and the biological activity of the protein drug is efficiently maintained. The method of loading the protein drug by the nanogel effectively solves the problems of short circulation time, poor endocytosis and easy inactivation and deterioration of the protein drug in vivo, and has wide development prospect in the aspect of realizing the in vivo application of the protein drug, such as the fields of tumor treatment and the like.
According to the preparation method of the protein drug-loaded composite material, the cross-linked grid is formed through the interaction of the host and the guest, and the protein drug is encapsulated in the nanogel, so that the protein drug can effectively circulate in vivo for a long time, is enriched at a tumor part and slowly releases the protein drug, and the capability of the protein drug taken up by cells is greatly enhanced. The preparation method provided by the invention is simple, the raw material source is wide, the batch production can be realized, and the industrialization can be realized.
The experimental result shows that the polyglutamic acid grafted polyethylene glycol grafted aminoazobenzene prepared by the invention and the polyglutamic acid grafted polyethylene glycol grafted amino β cyclodextrin carry the RNA enzyme, the size of the obtained RNA enzyme loaded nanogel is between 100 and 200nm, on a cell level, the endocytosis of the RNA enzyme loaded nanogel or pure RNA enzyme by the cell is compared, and the result shows that the endocytosis efficiency of the RNA enzyme is improved by more than two times by the nanogel.
For further illustration of the present invention, the polymer, polymer material, protein drug-loaded composite material and application provided by the present invention will be described in detail with reference to the following examples, but it should be understood that these examples are carried out on the premise of the technical solution of the present invention, and the detailed embodiments and specific procedures are given only for further illustration of the features and advantages of the present invention, not for limitation of the claims of the present invention, and the scope of protection of the present invention is not limited to the following examples.
Example 1
42.1g (160.0mmol) of gamma-benzyl-L-glutamate-N-dicarboxylic anhydride monomer (BLG-NCA) is dissolved in 270mL of anhydrous N, N-Dimethylformamide (DMF), and after stirring and dissolving, 1.0mL (1.0mmol/L DMF solution) of N-hexylamine (N-HA) is added, sealed, and stirred and reacted for 72 hours at the temperature of 25 ℃. After the reaction is finished, the obtained reaction solution is settled into 2.0L of diethyl ether, and the intermediate product poly (gamma-benzyl-L-glutamate) (PBLG) is obtained after the reaction solution is sequentially filtered, washed by the diethyl ether and dried for 24 hours in vacuum at room temperature.
10.0g of the poly (. gamma. -benzyl-L-glutamate) prepared above was dissolved in 100mL of dichloroacetic acid, and 30mL of a 33% hydrogen bromide/glacial acetic acid solution was added under stirring, followed by reaction under stirring at 30 ℃ for 1 hour. Then, the obtained reaction solution was settled in 1.0L of diethyl ether, centrifuged, and the obtained precipitate was redissolved with DMF, dialyzed with deionized water, and lyophilized to obtain poly (L-glutamic acid) homopolymer (PLG).
The poly (L-glutamic acid) homopolymer prepared in example 1 of the present invention was subjected to NMR analysis using deuterated water as a deuterated reagent.
The results showed that chemical shifts 4.43ppm were the signal peak for the last methyl group on the backbone, 2.21ppm was the signal peak for the methylene group attached to the carbonyl group on the pendant group, and chemical shifts 1.91ppm and 1.71ppm were the signal peaks for the methylene group attached to the backbone on the pendant group. According to nuclear magnetic calculation, the polymerization degree of the obtained poly (L-glutamic acid) was 150, and the overall yield was 80.7%.
Example 2
42.1g (160.0mmol) of gamma-benzyl-L-glutamate-N-dicarboxylic anhydride monomer (BLG-NCA) is dissolved in 270mL of anhydrous N, N-Dimethylformamide (DMF), and after stirring and dissolving, 1.0mL (1.0mmol/L DMF solution) of N-hexylamine (N-HA) is added, sealed, and stirred and reacted for 72 hours at the temperature of 25 ℃. Thereafter, 2.0g (20.0mmol) of acetic anhydride was added to the above reaction system, and the reaction was continued for 6 hours. After the reaction is finished, the obtained reaction solution is settled into 2.0L of diethyl ether, and the intermediate product poly (gamma-benzyl-L-glutamate) (PBLG) is obtained after the reaction solution is sequentially filtered, washed by the diethyl ether and dried for 24 hours in vacuum at room temperature.
10.0g of the poly (. gamma. -benzyl-L-glutamate) prepared above was dissolved in 100mL of dichloroacetic acid, and 30mL of a 33% hydrogen bromide/glacial acetic acid solution was added under stirring, followed by reaction under stirring at 30 ℃ for 1 hour. Then, the resulting reaction solution was settled into 1.0L of diethyl ether, centrifuged, and the resulting precipitate was redissolved with DMF, dialyzed with deionized water, and lyophilized to give acetyl-terminated poly (L-glutamic acid) homopolymer (PLG).
Example 3
To a dry reaction flask, poly (L-glutamic acid) (1.7g, 13.2mmol of glutamic acid unit) prepared in example 1, 3.5g (79.5mmol of ethylene glycol unit) of polyethylene glycol monomethyl ether (5000Da) were added, and 150mL of DMF was further added to dissolve the mixture. Then, 178mg (1.4mmol) of N, N-Diisopropylcarbodiimide (DIC) and 196mg (1.6mmol) of 4-Dimethylaminopyridine (DMAP) were added thereto, the mixture was sealed at 25 ℃ for reaction, after 48 hours, the resulting reaction solution was precipitated with 1.0L of diethyl ether, the resulting solid was redissolved with DMF, dialyzed with deionized water for 3 days, and lyophilized to obtain poly-L-glutamic acid to which polyethylene glycol was grafted.
The poly-L-glutamic acid grafted with polyethylene glycol obtained in example 3 of the present invention was subjected to nuclear magnetic resonance analysis using deuterated water as a solvent.
Referring to FIG. 1, FIG. 1 shows the preparation of polyethylene glycol grafted poly-L-glutamic acid prepared in example 31HNMR map.
As can be seen from the figure, the peak positions include: delta 4.25ppm (t, -CH)<),3.63ppm(t,-CH2CH2O-),3.31ppm(s,-OCH3),2.18ppm(m,-CH2COOH),1.96and1.83ppm(m,>CHCH2-),1.10–1.02ppm(m,-CH2CH2-),0.78ppm(t,-CH2-CH3)。
Example 4
Preparation of polyglutamic acid grafted polyethylene glycol grafted azobenzene
To three dry reaction bottles, polyglutamic acid-grafted polyethylene glycol (1000mg) prepared in example 3 was added, dissolved in 50ml of dry N, N-dimethylformamide, followed by addition of N, N-diisopropylethylamine (933mg) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (550mg), and the mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours in an oil bath at 40 ℃. Thereafter, aminoazobenzene (285mg) was added to the reaction solution, and the reaction was continued at room temperature for 48 hours. After the reaction is finished, the reaction solution is settled by ether, filtered, the solid is collected and dried in vacuum at room temperature, DMF redissolved solid is ultrafiltered by deionized water and freeze-dried to obtain polyglutamic acid grafted polyethylene glycol grafted azobenzene, and the yield is weighed and calculated.
The azobenzene polymer obtained in example 4 of the present invention was subjected to nuclear magnetic resonance analysis, and trifluoroacetic acid was used as a deuteration reagent.
See fig. 2. FIG. 2 is the NMR spectrum of polyglutamic acid grafted PEG-grafted azobenzene prepared in example 4.
From FIG. 2, significant characteristic azobenzene peaks (7.90ppm,8.00ppm,8.30ppm,8.50ppm) were observed, indicating that azobenzene was successfully bonded to the polymer.
Example 5
Preparation of polyglutamic acid grafted polyethylene glycol grafted amino beta-cyclodextrin
1000mg of polyglutamic acid grafted polyethylene glycol prepared in example 4 was added to each of the dried reaction bottles, and dissolved in 50ml of dried N, N-dimethylformamide, followed by addition of N, N-diisopropylethylamine (933mg) and 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (550mg), and the resulting mixture was placed in an oil bath at 40 ℃ and stirred at room temperature under a nitrogen atmosphere for 2 hours. Aminocyclodextrin (1776mg) was then added and the reaction was continued at room temperature for 48 h. And (3) settling the ethyl ether, filtering, collecting a solid, drying the solid at room temperature in vacuum, redissolving the solid by using N, N-dimethylformamide, performing ultrafiltration by using deionized water, and performing freeze-drying to obtain the polyglutamic acid grafted polyethylene glycol grafted amino cyclodextrin.
The polymer obtained in example 5 of the present invention was analyzed by nmr with deuterated water as a deuterated reagent.
See fig. 3. FIG. 3 is the NMR spectrum of polyglutamic acid grafted PEG-aminocyclodextrin prepared in example 5.
By comparison with FIG. 1, distinct peaks characteristic of cyclodextrins (3.39ppm,3.72ppm,4.86ppm) were observed, indicating that aminocyclodextrins were successfully bound to macromolecules.
Examples 6 to 7
Protein drug RNAse and interleukin 2 carrying
To 2 dry bottles were added glutamic acid-grafted polyethylene glycol-grafted aminoazobenzene prepared in example 4 (25mg) and glutamic acid-grafted polyethylene glycol-grafted aminocyclodextrin prepared in example 5 (25mg), respectively, and dissolved with 2ml of a PB (pH 7.4) solution, respectively.
The fluorescein-labeled protein drug rnase (15mg) or interleukin 2(15mg) was dissolved in 1ml of PB (pH 7.4) solution, and the three were mixed and vortexed for 20 seconds, followed by stirring at room temperature for 24 hours. The free RNase or interleukin 2 was removed by dialysis. The content of the supported RNase or interleukin 2 was measured by a fluorescence spectrophotometer method. The results showed that the excitation wavelength of the fluorescence spectrophotometer was 488nm, and the absorption wavelength was 518 nm.
And (3) carrying out dynamic light scattering analysis on the polymer RNA enzyme loaded nanogel obtained in the step, and determining the hydrodynamic radius of the nano particles formed by self-assembly in water.
Referring to FIG. 4, FIG. 4 is the result of dynamic light scattering in water at a concentration of 0.2mg/mL in the RNase-supported nanogel prepared in example 6 of the invention.
As can be seen from FIG. 4, the hydrodynamic radius of the self-assembled micelle is between 100 nm and 120nm, and the particle size distribution is uniform.
Example 8
Comparison of cellular endocytosis of RNAse-loaded nanogels with that of pure RNAse
The ability of the nanogel-loaded rnase to endocytosis of pure rnase by cells was examined by flow analysis. Six-well plates were plated with 40 million 4T1 cells in log phase growth per well and after cells were attached, the medium was replaced with a solution containing rnase or pure rnase loaded by nanogels. After endocytosis for 3h or 8h, the cells were washed 3 times with 1mL PBS solution, then digested for 1min with pancreatin without EDTA, and then blown to collect the cells. After centrifugation at 1000r for 5min, the supernatant was discarded to retain the cell pellet, and after repeated washing three times with 1mL of water, the cells were resuspended in 0.3mL of PBS. The sample was examined using a BDFACSVerse flow meter.
Referring to FIG. 5, FIG. 5 is a graph comparing the flow results obtained in example 8 of the present invention.
As can be seen in fig. 5, the nanogel significantly enhanced the ability of rnase to be endocytosed by cells.
Example 9
Pharmacokinetics of nanogel-loaded rnases and pure rnases
Wistar rats (weight 200-220 g) were taken 6 and randomly divided into two groups. The RNase and the pure RNase (the RNase is labeled with Cy5, and the concentration of the RNase and the pure RNase are 4mg kg) loaded by the nanogel through tail vein injection-1Based on RNase-Cy 5). Blood samples were collected from the orbital cavity at pre-designed time points (5, 30, 60, 120, 240, 480, 720 minutes). The samples were heparinized, serum was collected by centrifugation, and the concentration of RNase-Cy5 was measured by a fluorescence spectrophotometer (λ ex 649nm,λ em 670 nm). The half-life of the drug was calculated using PKSolver.
Referring to FIG. 6, FIG. 6 is a pharmacokinetic profile measured in example 9 of the present invention.
From FIG. 6 it can be seen that the nanogel greatly prolongs the circulation time of RNase in vivo. The result shows that the nanogel provided by the invention greatly improves the in vivo application rate of protein drugs, and the in vivo half-life period of the RNase is improved from 3.17h of the half-life period of pure RNase to 6.69h of the half-life period of the nanoRNase, so that the nanogel has great potential in the aspect of in vivo delivery of the protein drugs.
While the present invention has been described in detail with reference to a polymer, a nanogel for supporting protein drugs, and applications thereof, the principles and embodiments of the present invention are illustrated herein using specific examples, which are set forth merely to facilitate an understanding of the methods of the present invention and their core concepts, including the best mode, and to enable any person skilled in the art to practice the invention, including making and using any devices or systems and performing any combination thereof. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention. The scope of the invention is defined by the claims and may include other embodiments that occur to those skilled in the art. Such other embodiments are intended to be within the scope of the claims if they have structural elements that do not differ from the literal language of the claims, or if they include equivalent structural elements with insubstantial differences from the literal languages of the claims.
Claims (10)
1. A polymer, characterized in that the polymer has a structure represented by formula (I);
wherein R includes R1Or R2；
The R is1Selected from compounds having a hydrophobic internal cavity;
the R is2Selected from compounds that can intercalate into hydrophobic lumens;
L1、L2and L3Each independently selected from-CH2-or-CH2CH2-；
x, y and z are integers, x is more than or equal to 0, y is more than 0, z is more than 0, and x + y + z is more than or equal to 10 and less than or equal to 5000;
n is an integer, and n is more than or equal to 10 and less than or equal to 500.
2. The polymer of claim 1, wherein the compound having a hydrophobic internal cavity comprises one or more of a cyclodextrin, a calixarene, and a cucurbituril;
the compound capable of being embedded into the hydrophobic inner cavity comprises one or more of azobenzene, adamantane and biphenyl;
x + y + z is more than or equal to 30 and less than or equal to 300;
the R is1And R2Has the function of realizing the cross-linking of supermolecule effect.
3. A polymer material is characterized in that the polymer material is obtained by crosslinking a polymer with a structure shown in a formula (II) and a polymer with a structure shown in a formula (III);
wherein R is1Selected from compounds having a hydrophobic internal cavity;
R2selected from compounds that can intercalate into hydrophobic lumens;
L1、L2and L3Each independently selected from-CH2-or-CH2CH2-；
x, y and z are integers, x is more than or equal to 0, y is more than 0, z is more than 0, and x + y + z is more than or equal to 10 and less than or equal to 5000;
n is an integer, and n is more than or equal to 10 and less than or equal to 500.
4. The polymeric material of claim 3, wherein the cross-link is R in a polymer having a structure represented by formula (II)1And R in the polymer having a structure represented by the formula (III)2Cross-linking by supramolecular interaction;
the compound with a hydrophobic inner cavity comprises one or more of cyclodextrin, calixarene and cucurbituril;
the compound which can be embedded into the hydrophobic inner cavity comprises one or more of azobenzene, adamantane and biphenyl.
5. The polymeric material of claim 3, wherein after the cross-linking, the compound having a hydrophobic inner cavity in the polymer having the structure of formula (II) and the compound capable of being inserted into the hydrophobic inner cavity in the polymer having the structure of formula (III) are physically cross-linked by host-guest interaction to form a supramolecular network;
x + y + z is more than or equal to 30 and less than or equal to 300;
the polymer material has the function of carrying the drug.
6. A protein drug-loaded composite material, comprising the polymer material according to any one of claims 3 to 5 and a protein drug.
7. The composite of claim 6, wherein the protein drug comprises one or more of a cytokine, a chemokine, a monoclonal antibody and fragments thereof, and a therapeutic protein drug;
the molar ratio of the protein drug to the polymer material is 1: (0.05-20);
the protein drug is encapsulated in a cross-linked network of cross-linked meshes formed by the polymer material;
the composite material includes a nanomaterial.
8. The composite material of claim 6, wherein the composite material has a size of 50 to 1000 nm;
the composite material comprises a nanogel;
the protein drug is encapsulated inside the nanogel;
the protein medicine comprises one or more of RNase, interleukin 2, interleukin 12 and cytochrome C;
the protein drug-loaded composite material is obtained by mixing and reacting a polymer solution with a structure shown in a formula (II), a polymer solution with a structure shown in a formula (III) and a protein drug.
9. The composite material of claim 8, wherein the solution comprises a buffer solution;
the mixing reaction time is 2-48 h;
the molar ratio of the polymer having the structure represented by the formula (II) to the polymer having the structure represented by the formula (III) is 1: (0.1 to 20);
the molar ratio of the protein drug to the polymer having the structure represented by formula (II) is 1: (0.1 to 20);
the concentration of the polymer solution with the structure shown in the formula (II) is 0.01-10 mM;
the concentration of the polymer solution with the structure shown in the formula (III) is 0.01-10 mM.
10. Use of a polymer according to any one of claims 1 to 2, a polymer material according to any one of claims 3 to 5 or a protein-loaded drug composite according to any one of claims 6 to 9 in the field of tumor therapy, metabolic disorders and autoimmune diseases.
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