CN108752541B - Amphiphilic block polymer with hypoxia and temperature dual responsiveness and taking azo bond as connecting bond and preparation method thereof - Google Patents
Amphiphilic block polymer with hypoxia and temperature dual responsiveness and taking azo bond as connecting bond and preparation method thereof Download PDFInfo
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- 206010021143 Hypoxia Diseases 0.000 title claims abstract description 22
- 230000004043 responsiveness Effects 0.000 title claims abstract description 21
- 230000009977 dual effect Effects 0.000 title claims abstract description 16
- 229920000642 polymer Polymers 0.000 title claims abstract description 15
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 230000007954 hypoxia Effects 0.000 title claims description 16
- -1 2-methoxyethoxy Chemical group 0.000 claims abstract description 19
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000001301 oxygen Substances 0.000 claims abstract description 17
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 17
- 239000002202 Polyethylene glycol Substances 0.000 claims abstract description 15
- 229920001223 polyethylene glycol Polymers 0.000 claims abstract description 15
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- 238000000034 method Methods 0.000 claims abstract description 8
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- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 36
- 229910052757 nitrogen Inorganic materials 0.000 claims description 36
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 35
- 239000000047 product Substances 0.000 claims description 31
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- 239000000243 solution Substances 0.000 claims description 29
- 239000002904 solvent Substances 0.000 claims description 28
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 27
- 238000006243 chemical reaction Methods 0.000 claims description 21
- 229910021589 Copper(I) bromide Inorganic materials 0.000 claims description 18
- NKNDPYCGAZPOFS-UHFFFAOYSA-M copper(i) bromide Chemical compound Br[Cu] NKNDPYCGAZPOFS-UHFFFAOYSA-M 0.000 claims description 18
- 239000000178 monomer Substances 0.000 claims description 18
- 239000003054 catalyst Substances 0.000 claims description 17
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 14
- 239000006227 byproduct Substances 0.000 claims description 14
- BEYOBVMPDRKTNR-UHFFFAOYSA-N chembl79759 Chemical compound C1=CC(O)=CC=C1N=NC1=CC=CC=C1 BEYOBVMPDRKTNR-UHFFFAOYSA-N 0.000 claims description 14
- 239000005457 ice water Substances 0.000 claims description 14
- 238000003828 vacuum filtration Methods 0.000 claims description 14
- 229910021591 Copper(I) chloride Inorganic materials 0.000 claims description 13
- OXBLHERUFWYNTN-UHFFFAOYSA-M copper(I) chloride Chemical compound [Cu]Cl OXBLHERUFWYNTN-UHFFFAOYSA-M 0.000 claims description 13
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- KAKZBPTYRLMSJV-UHFFFAOYSA-N Butadiene Chemical group C=CC=C KAKZBPTYRLMSJV-UHFFFAOYSA-N 0.000 claims description 8
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- 238000003756 stirring Methods 0.000 claims description 6
- ROFVEXUMMXZLPA-UHFFFAOYSA-N Bipyridyl Chemical group N1=CC=CC=C1C1=CC=CC=N1 ROFVEXUMMXZLPA-UHFFFAOYSA-N 0.000 claims description 4
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 claims description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 4
- 150000001412 amines Chemical class 0.000 claims description 4
- 125000000524 functional group Chemical group 0.000 claims description 4
- DWFKOMDBEKIATP-UHFFFAOYSA-N n'-[2-[2-(dimethylamino)ethyl-methylamino]ethyl]-n,n,n'-trimethylethane-1,2-diamine Chemical compound CN(C)CCN(C)CCN(C)CCN(C)C DWFKOMDBEKIATP-UHFFFAOYSA-N 0.000 claims description 4
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 claims description 3
- 229920001577 copolymer Polymers 0.000 claims description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 2
- 239000000693 micelle Substances 0.000 abstract description 10
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- PEXGTUZWTLMFID-UHFFFAOYSA-N 2-phenyldiazenylphenol Chemical compound OC1=CC=CC=C1N=NC1=CC=CC=C1 PEXGTUZWTLMFID-UHFFFAOYSA-N 0.000 abstract 1
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- 239000003999 initiator Substances 0.000 abstract 1
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- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 16
- 238000005303 weighing Methods 0.000 description 10
- 230000001146 hypoxic effect Effects 0.000 description 5
- 230000035484 reaction time Effects 0.000 description 4
- 229940079593 drug Drugs 0.000 description 3
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- 239000002861 polymer material Substances 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- ORILYTVJVMAKLC-UHFFFAOYSA-N adamantane Chemical compound C1C(C2)CC3CC1CC2C3 ORILYTVJVMAKLC-UHFFFAOYSA-N 0.000 description 2
- DMLAVOWQYNRWNQ-UHFFFAOYSA-N azobenzene Chemical compound C1=CC=CC=C1N=NC1=CC=CC=C1 DMLAVOWQYNRWNQ-UHFFFAOYSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229920005604 random copolymer Polymers 0.000 description 2
- HVXBOLULGPECHP-WAYWQWQTSA-N Combretastatin A4 Chemical compound C1=C(O)C(OC)=CC=C1\C=C/C1=CC(OC)=C(OC)C(OC)=C1 HVXBOLULGPECHP-WAYWQWQTSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- SUAKHGWARZSWIH-UHFFFAOYSA-N N,N‐diethylformamide Chemical compound CCN(CC)C=O SUAKHGWARZSWIH-UHFFFAOYSA-N 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229940009456 adriamycin Drugs 0.000 description 1
- 238000011319 anticancer therapy Methods 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 description 1
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- 229920001400 block copolymer Polymers 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
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- 229960005537 combretastatin A-4 Drugs 0.000 description 1
- HVXBOLULGPECHP-UHFFFAOYSA-N combretastatin A4 Natural products C1=C(O)C(OC)=CC=C1C=CC1=CC(OC)=C(OC)C(OC)=C1 HVXBOLULGPECHP-UHFFFAOYSA-N 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
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- 238000012377 drug delivery Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
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- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000010550 living polymerization reaction Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- QNILTEGFHQSKFF-UHFFFAOYSA-N n-propan-2-ylprop-2-enamide Chemical class CC(C)NC(=O)C=C QNILTEGFHQSKFF-UHFFFAOYSA-N 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F293/00—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule
- C08F293/005—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule using free radical "living" or "controlled" polymerisation, e.g. using a complexing agent
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F2438/00—Living radical polymerisation
- C08F2438/01—Atom Transfer Radical Polymerization [ATRP] or reverse ATRP
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- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Polyethers (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention belongs to the field of biological materials and nano materials, and relates to an amphiphilic block polymer which takes azo bonds as connecting bonds and has dual responsiveness of oxygen lack and temperature and a preparation method thereof. The method comprises the following specific steps: the method comprises the following steps of initiating atom transfer radical polymerization of 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) and oligo (ethylene glycol methyl ether) methacrylate by taking a hydroxy azobenzene and polyethylene glycol copolymer with one end esterified as an initiator to obtain the amphiphilic block copolymer material with an azo bond as a connecting bond. The amphiphilic block copolymer material prepared by the invention has dual responsiveness of oxygen deficiency and temperature, can be self-assembled in aqueous solution to form nano micelle, and can be applied to aspects of biomedicine, nano drug carriers, diagnosis and imaging and the like. The method is simple and feasible, and the raw materials can be industrially produced, so the method has wide application prospect.
Description
Technical Field
The invention belongs to the fields of high molecular materials and biomedical engineering, and particularly relates to an amphiphilic block polymer which takes an azo bond as a connecting bond and has dual responsiveness of oxygen lack and temperature and a preparation method thereof.
Background
The p-hydroxyazobenzene (4, 4' -Azobenzenedioxy) has a highly symmetrical chemical structure, the hydroxyl groups at two ends are used as two functional groups to respectively connect macromolecules with required performance, and the nitrogen-nitrogen double bond at the symmetrical center can be broken in an environment with low oxygen content. The novel amphiphilic block copolymer material prepared by taking azo bonds as hypoxic response functional groups draws great attention of people due to the unique preparation method and excellent performance. Kulkarni P and the like (Kulkarni P, Haldar M K, You S, et al. biomacromolecules,2016,17(8):2507.) synthesize a polymer which takes azo bonds as connecting bonds and has two ends respectively connected with polyethylene glycol and polylactic acid, and the polymer has hypoxia responsiveness and cell targeting recognition capability, can be used as a drug carrier and has wide application prospect in the field of biomedicine.
When the external environment changes slightly, some polymers can correspondingly and rapidly change relatively obviously physically or chemically, and the polymers are called environment-responsive polymers and are also called environment-sensitive polymers or stimulus-responsive polymers. In the past, general research on stimulus-responsive polymers has been put on stimulus-responsive factors such as temperature, pH, ultraviolet, magnetism, and the like. Patent application 201410209753.3 discloses a preparation method of a temperature-responsive magnetic resonance contrast supramolecular nano-micelle, wherein a supramolecular nano-micelle construction unit comprises three components, namely bridging trimeric b-cyclodextrin, polyethylene glycol modified metallomanganese porphyrin and adamantane modified poly-N-isopropylacrylamide, and the three components construct a supramolecular assembly through host-guest inclusion coordination interaction; the supermolecule nano micelle can be used for loading and controlling release of hydrophobic dye or drug molecules. Hypoxia, a stimulus response factor, is the most important indicator in tumor cells as a new stimulus, because only the change of hypoxia represents the growth process of tumor. Some new response materials developed for hypoxia, a new stimulus factor, have been reported. Liu H, etc. (Liu H, Zhang R, Niu Y, et al. RscAdvances,2015,5(27):20848 20857.) the polyethylene glycol-hexanethiol and combretastatin A-4 are connected through azobenzene to form a nano micelle, and the nano micelle wraps the adriamycin to construct a hypoxia response drug delivery system for anticancer therapy.
In summary, a single stimulus cannot be used as a characteristic index of a complex physiological system.
Disclosure of Invention
In order to solve the problems, the invention discloses an amphiphilic block polymer which takes azo bonds as connecting bonds and has dual responsiveness of oxygen lack and temperature and a preparation method thereof by combining with a tumor microenvironment, and the amphiphilic block polymer is used as a nano carrier for delivering anticancer drugs. Based on the advanced living polymerization method, azo bonds are used as hypoxic response groups, and atom transfer radical polymerization is combined, so that random copolymers of polyethylene glycol and 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) and oligoethylene glycol methyl ether methacrylate are respectively introduced into two ends of azobenzene, the random copolymers have low critical solution temperature, and therefore the amphiphilic block copolymer material with hypoxic and temperature dual responsiveness is prepared, can be self-assembled in an aqueous solution to form a nano micelle, and can be widely applied to the fields of biomedicine, nano drug carriers, diagnostic imaging and the like.
The amphiphilic block polymer with double responses of oxygen lack and temperature and taking an azo bond as a connecting bond is characterized by having the structural formula shown in the specification, wherein the molecular weight is 10000-30000, the left end is monomethoxy polyethylene glycol, the middle part is an azo bond oxygen lack response functional group, and the right end is a copolymer of temperature response 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) and oligoethylene glycol methyl ether methacrylate.
The invention provides a preparation method of amphiphilic block polymer with hypoxia and temperature dual responsiveness by taking azo bonds as connecting bonds, which is characterized by comprising the following steps in sequence: :
(1) dissolving 2-bromoisobutyric acid in a solvent A, gradually dropwise adding the solution into a solvent A solution of p-hydroxyazobenzene, N' -dicyclohexylcarbodiimide and a catalyst B, reacting at-10-5 ℃ for 10-60 minutes, stirring the obtained solution at room temperature for 12-36 hours, removing reaction byproducts through vacuum filtration, removing the solvent through rotary evaporation, purifying the product through column chromatography, selecting a mixed solution of diethyl ether and hexane as an eluent, collecting the product, and drying in a vacuum oven at 20-70 ℃ for 15-30 hours;
(2) dissolving the product obtained in the step (1), N' -dicyclohexylcarbodiimide and 4-dimethylaminopyridine in dichloromethane and dimethylformamide 1: 1-2, dropwise adding carboxylated monomethyl polyethylene glycol into the solution, reacting at room temperature for 16-26 hours under the protection of nitrogen, removing by-products by vacuum filtration, collecting the product, precipitating in a precipitator C, collecting the precipitate, and vacuum-drying;
(3) dissolving the product obtained in the step (2), 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) monomer, oligoethylene glycol methyl ether methacrylate monomer, complexing agent N, N, N' -pentamethyldiethylenetriamine and catalyst cuprous bromide in solvent anhydrous dimethylformamide, vacuumizing and removing oxygen in an ice water bath, introducing nitrogen, removing the ice water bath, adjusting the reaction temperature to be 60-100 ℃, reacting for 15-30 hours, cooling to room temperature, dialyzing and purifying the crude product, and freeze-drying to obtain the final product.
In the invention, the solvent A is one or more of dichloromethane, tetrahydrofuran, N-dimethylformamide, N-diethylformamide, N-dimethylacetamide and dimethyl sulfoxide; the catalyst B is one or more of cuprous bromide/bipyridyl, cuprous chloride/bipyridyl, cuprous bromide/pentamethyl divinyl triamine, cuprous chloride/pentamethyl divinyl triamine, cuprous bromide/tri (2-methylaminoethyl) amine, cuprous chloride/tri (2-methylaminoethyl) amine, cuprous bromide/hexamethyl triethylene tetramine, cuprous chloride/hexamethyl triethylene tetramine, cuprous bromide/2-pyridine formaldehyde normal propyl amine and cuprous chloride/2-pyridine formaldehyde normal propyl amine; the precipitator C is one or more of cyclohexane, n-hexane and diethyl ether.
Compared with the prior art, the invention has the beneficial effects that: the raw materials adopted by the invention are wide in source, the used p-hydroxyazobenzene, polyethylene glycol, 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) monomer, oligoethylene glycol methyl ether methacrylate monomer, catalyst, solvent and the like can be industrially produced, and the synthesis method is simple and easy to implement. In addition, all the raw materials have biocompatibility and biological nontoxicity. The synthesized amphiphilic block polymer material with double responsiveness of oxygen lack and temperature, which takes azo bonds as connecting bonds, has double responsiveness of oxygen lack and temperature, azo bonds are broken under the action of a hypoxic environment and enzyme catalysis, the responsiveness of oxygen lack is reflected, different molecular chain morphologies are presented along with the difference of environmental temperature, and the critical dissolution temperature can be regulated and controlled by changing the feeding ratio of a 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) monomer and an oligoethylene glycol methyl ether methacrylate monomer during synthesis. Due to the fact that polyethylene glycol has hydrophilicity and poly 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) -oligo (ethylene glycol methyl ether) methacrylate has hydrophobicity, the block copolymer material can self-assemble to form micelles in water, and the particle size of the micelles can be adjusted by changing the chain segment length of the copolymer. The amphiphilic block polymer with double responsiveness of oxygen lack and temperature, which takes azo bonds as connecting bonds, can be widely applied to the fields of biomedicine, nano drug carriers, diagnosis and imaging and the like.
Description of the drawings:
FIG. 1: the structural schematic diagram of an amphiphilic block polymer with hypoxia and temperature dual responsiveness, which is prepared in example 1 and takes azo bonds as connecting bonds.
FIG. 2: H-NMR chart of amphiphilic block polymer with hypoxia and temperature dual responsiveness prepared in example 1 and using azo bond as a connecting bond.
FIG. 3: the drug release efficiency chart of the amphiphilic block polymer with hypoxia and temperature dual responsiveness, which is prepared by the method in example 1 and takes azo bonds as connecting bonds.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to limit the scope of the present invention.
The molecular structure of the amphiphilic block polymer material with the azo bond as the connecting bond and with the hypoxia and temperature dual responsiveness is determined by a nuclear magnetic resonance analyzer (NMR).
Example 1:
(1) 5.4g of 2-bromoisobutyric acid was weighed out and dissolved in methylene chloride as a solvent, and this solution was gradually added dropwise to a methylene chloride solution of 7.5g of p-hydroxyazobenzene, 7.05g N, N' -dicyclohexylcarbodiimide, and 0.32g of dimethylformamide as a catalyst at a reaction temperature of 0 ℃ for 30 minutes, and the resulting solution was stirred at room temperature. The reaction by-products were removed by vacuum filtration and the solvent was removed by rotary evaporation. Purifying the product by column chromatography, selecting a mixed solution of diethyl ether and hexane as an eluent, collecting the product, and drying in a vacuum oven at 50 ℃ for 15 h;
(2) 1.12g of p-hydroxyazobenzene, 0.65g N, N' -dicyclohexylcarbodiimide, 0.13g of 4-dimethylaminopyridine were weighed out and dissolved in dichloromethane and dimethylformamide 1: 1, 2.004g of carboxylated monomethoxy polyethylene glycol is weighed and added into the solution dropwise, the reaction is carried out under the protection of nitrogen, the reaction temperature is room temperature, and the reaction time is 24 hours. Removing the by-product by vacuum filtration, collecting the product, precipitating in ether as a precipitator, collecting the precipitate, and vacuum drying;
(3) weighing 2.5g of the product obtained in the step (2), 7.9g of 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) monomer, 1.76g of oligoethylene glycol methyl ether methacrylate monomer, 0.283g of complexing agent N, N, N' -pentamethyldiethylenetriamine and 0.19g of cuprous bromide catalyst, dissolving in anhydrous dimethylformamide as a solvent, vacuumizing and removing oxygen in an ice-water bath, introducing nitrogen, removing the ice-water bath, adjusting the reaction temperature to 100 ℃, reacting for 30 hours, cooling to room temperature, dialyzing and purifying the crude product, and freeze-drying to obtain the final product.
The H-NMR chart of an amphiphilic block polymer with hypoxia and temperature dual responsiveness by taking an azo bond as a connecting bond is shown in figure 2.
Example 2:
(1) weighing 5.0g of 2-bromoisobutyric acid, dissolving in tetrahydrofuran solvent, gradually dropwise adding the solution into tetrahydrofuran solvent solution of 7.2g of p-hydroxyazobenzene, 6.98g N, N' -dicyclohexylcarbodiimide and 0.29g of cuprous chloride catalyst, stirring the obtained solution at room temperature for 12 hours at the reaction temperature of 5 ℃, removing reaction byproducts through vacuum filtration, removing the solvent through rotary evaporation, purifying the product through column chromatography, selecting a mixed solution of diethyl ether and hexane as eluent, collecting the product, and drying in a vacuum oven at the temperature of 70 ℃ for 15 hours;
(2) 0.93g of p-hydroxyazobenzene, 0.54g N, N' -dicyclohexylcarbodiimide, 0.09g of 4-dimethylaminopyridine were weighed out and dissolved in dichloromethane and dimethylformamide 1: 2, weighing 1.84g of carboxylated monomethoxy polyethylene glycol, dropwise adding the weighed monomethoxy polyethylene glycol into the solution, reacting at room temperature under the protection of nitrogen for 16 hours, removing by-products by vacuum filtration, collecting the product, precipitating in a precipitator, namely diethyl ether, collecting the precipitate, and drying in vacuum;
(3) weighing 2.0g of the product obtained in the step (2), 7.54g of 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) monomer, 1.72g of oligoethylene glycol methyl ether methacrylate monomer, 0.376g of complexing agent N, N, N' -pentamethyldiethylenetriamine and 0.17g of cuprous bromide catalyst, dissolving the raw materials in anhydrous dimethylformamide as a solvent, vacuumizing and removing oxygen in an ice-water bath, introducing nitrogen, removing the ice-water bath, adjusting the reaction temperature to 80 ℃, reacting for 15 hours, cooling to room temperature, dialyzing and purifying the crude product, and freeze-drying to obtain the final product.
Example 3:
(1) weighing 4.8g of 2-bromoisobutyric acid, dissolving the 2-bromoisobutyric acid in a solvent N, N-dimethylformamide, gradually dropwise adding the solution into a solvent N, N-diethylformamide solution of 6.8g of p-hydroxyazobenzene, 6.65g N, N' -dicyclohexylcarbodiimide and 0.26g of cuprous chloride serving as a catalyst, wherein the reaction temperature is 5 ℃, the reaction time is 40 minutes, stirring the obtained solution at room temperature for 26 hours, removing reaction byproducts through vacuum filtration, removing the solvent through rotary evaporation, purifying the product through column chromatography, selecting a mixed solution of diethyl ether and hexane as an eluent, collecting the product, and drying the product in a vacuum oven at 50 ℃ for 24 hours;
(2) 1.58g of p-hydroxyazobenzene, 0.72g N, N' -dicyclohexylcarbodiimide, 0.21g of 4-dimethylaminopyridine were weighed out and dissolved in dichloromethane and dimethylformamide 1: 2, dropwise adding 2.14g of carboxylated monomethoxy polyethylene glycol into the solution, reacting at room temperature for 20 hours under the protection of nitrogen, removing by-products by vacuum filtration, collecting the product, precipitating in a precipitator cyclohexane, collecting the precipitate, and drying in vacuum;
(3) weighing 2.4g of the product obtained in the step (2), 6.89g of 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) monomer, 1.65g of oligoethylene glycol methyl ether methacrylate monomer, 0.32g of complexing agent N, N, N' -pentamethyldiethylenetriamine and 0.18g of cuprous bromide catalyst, dissolving in solvent anhydrous dimethylformamide, vacuumizing and removing oxygen in an ice-water bath, introducing nitrogen, removing the ice-water bath, adjusting the reaction temperature to 100 ℃, reacting for 15 hours, cooling to room temperature, dialyzing and purifying the crude product, and freeze-drying to obtain the final product.
Example 4:
(1) weighing 4.6g of 2-bromoisobutyric acid, dissolving in a solvent N, N-dimethylacetamide, gradually dropwise adding the solution into a solvent N, N-dimethylacetamide solution of p-hydroxyazobenzene, N' -dicyclohexylcarbodiimide and cuprous chloride serving as a catalyst, wherein the reaction temperature is 5 ℃, the reaction time is 30 minutes, stirring the obtained solution at room temperature for 20 hours, removing reaction byproducts through vacuum filtration, removing the solvent through rotary evaporation, purifying the product through column chromatography, selecting a mixed solution of diethyl ether and hexane as an eluent, collecting the product, and drying in a vacuum oven at 45 ℃ for 30 hours;
(2) 1.84g of p-hydroxyazobenzene, 0.82g N, N' -dicyclohexylcarbodiimide, 0.24g of 4-dimethylaminopyridine were weighed out and dissolved in dichloromethane and dimethylformamide 1: 1.5, dropwise adding 2.23g of carboxylated monomethoxy polyethylene glycol into the solution, reacting at room temperature under the protection of nitrogen for 26 hours, removing by-products by vacuum filtration, collecting products, precipitating in a precipitator, namely diethyl ether, collecting precipitates, and drying in vacuum;
(3) weighing 2.87g of the product obtained in the step (2), 8.04g of 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) monomer, 2.14g of oligoethylene glycol methyl ether methacrylate monomer, 0.412g of complexing agent N, N, N' -pentamethyldiethylenetriamine and 0.20g of cuprous bromide catalyst, dissolving in anhydrous dimethylformamide as a solvent, vacuumizing and removing oxygen in an ice-water bath, introducing nitrogen, removing the ice-water bath, adjusting the reaction temperature to 100 ℃, reacting for 24 hours, cooling to room temperature, dialyzing and purifying the crude product, and freeze-drying to obtain the final product.
Example 5:
(1) weighing 5.8g of 2-bromoisobutyric acid, dissolving in a solvent dichloromethane, gradually dropwise adding the solution into a solvent dimethyl sulfoxide solution of p-hydroxyazobenzene, N' -dicyclohexylcarbodiimide and cuprous bromide serving as a catalyst, wherein the reaction temperature is 0 ℃, the reaction time is 60 minutes, stirring the obtained solution at room temperature for 36 hours, removing reaction byproducts through vacuum filtration, removing the solvent through rotary evaporation, purifying the product through column chromatography, selecting a mixed solution of diethyl ether and hexane as an eluent, collecting the product, and drying in a vacuum oven at 70 ℃ for 20 hours;
(2) 1.94g of p-hydroxyazobenzene, 0.91g N, N' -dicyclohexylcarbodiimide, 0.36g of 4-dimethylaminopyridine were weighed out and dissolved in dichloromethane and dimethylformamide 1: 1, dropwise adding 2.4g of carboxylated monomethyl polyethylene glycol into the solution, reacting at room temperature under the protection of nitrogen for 26 hours at room temperature under the protection of nitrogen, removing by-products by vacuum filtration, collecting the product, precipitating in a precipitator, namely n-hexane, collecting the precipitate, and drying in vacuum;
(3) weighing 2.61g of the product obtained in the step (2), 8.12g of 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) monomer, 2.26g of oligoethylene glycol methyl ether methacrylate monomer, 0.45g of complexing agent N, N, N' -pentamethyldiethylenetriamine and 0.24g of cuprous bromide catalyst, dissolving in anhydrous dimethylformamide as a solvent, vacuumizing and removing oxygen in an ice-water bath, introducing nitrogen, removing the ice-water bath, adjusting the reaction temperature to 70 ℃, reacting for 30 hours, cooling to room temperature, dialyzing and purifying the crude product, and freeze-drying to obtain the final product.
The structural formula of the amphiphilic block polymer material with hypoxia and temperature dual responsiveness, which takes azo bonds as connecting bonds, is shown in figure 1. In a hypoxic environment with the temperature of lower than 33-37 ℃, the micelle is dissociated, the drug is effectively released, and the release rate is 90-96% after 12 hours, as shown in figure 3.
The above description is only illustrative of the preferred embodiments of the present invention and should not be taken as limiting the scope of the invention in any way. Any changes or modifications made by those skilled in the art based on the above disclosure should be considered as equivalent effective embodiments, and all the changes or modifications should fall within the protection scope of the technical solution of the present invention.
Claims (3)
1. An amphiphilic block polymer with hypoxia and temperature dual responsiveness by taking an azo bond as a connecting bond is characterized in that the structural formula is shown as follows, the molecular weight is 10000-30000, the left end is monomethoxy polyethylene glycol, the middle part is an azo bond hypoxia response functional group, and the right end is a copolymer of temperature responsiveness 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) ethyl ester and oligoethylene glycol monomethyl ether methacrylate;
2. a method for preparing the amphiphilic block polymer with hypoxia and temperature dual responsiveness using azo bond as a connecting bond according to claim 1, which is characterized by comprising the following steps in sequence:
(1) dissolving 2-bromoisobutyric acid in a solvent A, gradually dropwise adding the solution into a solvent A solution of p-hydroxyazobenzene, N' -dicyclohexylcarbodiimide and a catalyst B, reacting at-10-5 ℃ for 10-60 minutes, stirring the obtained solution at room temperature for 12-36 hours, removing reaction byproducts through vacuum filtration, removing the solvent through rotary evaporation, purifying the product through column chromatography, selecting a mixed solution of diethyl ether and hexane as an eluent, collecting the product, and drying in a vacuum oven at 20-70 ℃ for 15-30 hours;
(2) dissolving the product obtained in the step (1), N' -dicyclohexylcarbodiimide and 4-dimethylaminopyridine in dichloromethane and dimethylformamide 1: 1-2, dropwise adding carboxylated monomethyl polyethylene glycol into the solution, reacting at room temperature for 16-26 hours under the protection of nitrogen, removing by-products by vacuum filtration, collecting the product, precipitating in a precipitator C, collecting the precipitate, and vacuum-drying;
(3) dissolving the product obtained in the step (2), a 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) ethyl ester monomer, an oligoethylene glycol methyl ether methacrylate monomer, a complexing agent N, N, N' -pentamethyldiethylenetriamine and a catalyst cuprous bromide in a solvent of anhydrous dimethylformamide, vacuumizing and removing oxygen in an ice-water bath, introducing nitrogen, removing the ice-water bath, adjusting the reaction temperature to be 60-100 ℃, reacting for 15-30 hours, cooling to room temperature, dialyzing and purifying the crude product, and freeze-drying to obtain the final product.
3. The preparation method according to claim 2, wherein the solvent A is one or more of dichloromethane, tetrahydrofuran, N-dimethylformamide, N-diethylformamide, N-dimethylacetamide and dimethyl sulfoxide; the catalyst B is one or more of cuprous bromide/bipyridyl, cuprous chloride/bipyridyl, cuprous bromide/pentamethyl divinyl triamine, cuprous chloride/pentamethyl divinyl triamine, cuprous bromide/tri (2-methylaminoethyl) amine, cuprous chloride/tri (2-methylaminoethyl) amine, cuprous bromide/hexamethyl triethylene tetramine, cuprous chloride/hexamethyl triethylene tetramine, cuprous bromide/2-pyridine formaldehyde normal propyl amine and cuprous chloride/2-pyridine formaldehyde normal propyl amine; the precipitator C is one or more of cyclohexane, n-hexane and diethyl ether.
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