Preparation method and application of nanogel with hunger combined gas therapy anti-tumor function
A nanogel and gas therapy technology, applied in the field of material and biomedical preparation, to achieve the effects of enhancing stability, improving encapsulation efficiency, and being difficult to dissociate
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Embodiment 1
[0034] The synthesis of embodiment 1 PEI-AATA-Arg
[0035] (1) Synthesis of azidated polyethyleneimine PEI-AATA
[0036] The synthetic route of PEI-AATA is as follows:
[0037]
[0038]Dissolve azidoacetic acid (48 μL, 0.64 mmol) in high-purity water, and then add catalyst 1-ethyl-(3-dimethylaminopropyl) carbodiimide N-hydroxysuccinimide to activate the carboxyl group ; At the same time, polyethyleneimine (300mg, 0.16mmol) was dissolved in high-purity water, concentrated hydrochloric acid was added to adjust the pH, and then activated azidoacetic acid solution was added. After 24 hours of reaction, it was dialyzed and freeze-dried to obtain light yellow powder azide Polyethyleneimine, whose structure is designated as PEI-AATA.
[0039] (2) Synthesis of functionalized polyethyleneimine containing azide group (PEI-AATA-Arg)
[0040] The synthetic route of PEI-AATA-Arg is as follows:
[0041]
[0042] Dissolve L-arginine (278mg, 1.6mmol) in high-purity water, add concen...
Embodiment 2
[0043] Synthesis of Example 2 BCN-MPA-PAC-PEG-PAC-MPA-BCN
[0044] (1) Synthesis of triblock copolymer PAC-PEG-PAC containing acrylate functional groups
[0045] In the glove box, 500 mg PEG and 120 mg AC monomer were dissolved in dichloromethane and added to a sealed reactor, then a catalytic amount of bis(bistrimethylsilyl)amine zinc was added, and the reactor was sealed and transferred out In a glove box, put it in a 40°C oil bath to react for 24 hours, stop the reaction with glacial acetic acid, precipitate in glacial ether, and finally centrifuge, discard the supernatant and dry it in vacuum to obtain the product. The NMR results show that AC in the triblock copolymer The proportion of the unit is 16.6%, and its structure is designated as PAC-PEG-PAC.
[0046] (2) The double bonds of the AC units of the triblock copolymer are completely replaced by carboxyl groups
[0047] Dissolve 0.1 g of triblock copolymer PAC-PEG-PAC in dimethylformamide (DMF), add mercaptopropionic...
Embodiment 3
[0050] The preparation of embodiment 3 nanogels
[0051] (1) Preparation of nanozyme complex
[0052] Glucose oxidase Gox and PEI-AATA-Arg were respectively dissolved in high-purity water at a concentration of 1 mg / mL, the two solutions were vortexed and mixed in equal proportions and then left to stand to obtain the nanozyme complex. After fine-tuning the pH of the PEI-AATA-Arg solution with sodium hydroxide, the two solutions in different proportions can be mixed to obtain a nanoenzyme complex with uniform particle size, with an average particle size of 190 nm and a particle size distribution of 0.18. This nanozyme complex is stable on rest, but dissociates in PBS.
[0053] (2) Preparation of nanogel
[0054] Dissolve BCN-MPA-PAC-PEG-PAC-MPA-BCN in dimethyl sulfoxide, and slowly add it dropwise to the nanocomposite obtained in (1) under stirring (equal molar azide group and alkynyl group amount), stirred for a period of time to generate cross-linking, and finally removed ...
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