Synthesis And Application Of New Protein Delivery Material Based On ?-cyclodextrin

Cyclodextrin(CD)is an oligosaccharide consisting of seven D-glucose units produced by enzymatic degradation of starch.It has a unique cylindrical hydrophobic cavity and a hydrophilic outer structure,For a variety of hydrophobic molecules have a good package and performance.?-cyclodextrin is the second generation of supramolecular macrocyclic compounds following crown ethers.In the field of pharmacy,?-cyclodextrin can contain a variety of hydrophobic drugs through the host-guest interaction,and improve the solubility of the drug,stability,and bioavailability.However,the natural ?-cyclodextrin is poor in water solubility and has a single functional group,which limits its application in the field of drug delivery.In this study,?-cyclodextrin was used to synthesize the structure of ?-cyclodextrin,which was structured by "Click reaction".The ability of self-assembly of complex proteins in water was studied,and the ability to deliver proteins into cells as a protein transport material was studied.The main work is as follows:1.?-Cyclodextrin Main Modification: The ?-cyclodextrin is modified with a synthetic hydrophobic single strand linked by a "Click reaction".The macromolecule with amphiphilic can be self-assembly in water,compound protein.The macromolecule can be combined with the target designed by the host-guest response to serve as a transport protein.The terminal of the macromolecule contains an azide group which can react with the triple bond to contain a crosslinking agent containing a specific reactive functional group.2.Targeted synthesis: Studies have shown that adamantane(AMD)is the best molecule with ?-cyclodextrin host-guest inclusion.For the adamantane modification,the multi-targeting intermediates were synthesized by simple amide reaction and Michael addition reaction.Synthesis of AMD-PEG-FA,AMD-PEG-Gal,water-soluble target molecules,and ?-cyclodextrin host molecules can be self-assembly in the water host-guest effect inclusion.3.Preparation of cross-linking agent: First,a compound molecule capable of reacting with an oxygen free radical is synthesized and then a special triple bond compound molecule is synthesized.It can be combined with the self-assembly of the modified reaction of the azo-containing molecule with the modified reaction of the ?-cyclodextrin.A water-soluble model cross-linker molecule was synthesized by synthesizing a specific triple bond compound with PEG 1000 for subsequent cell experiments.4.Biological properties of the experiment: The different mass of the cyclodextrin host material was combined with the quantitative BSA protein in water and then loaded onto the agarose gel electrophoresis.The results showed that the material could be compounded with the protein and found the lowest mass ratio.The synthesized compounds were self-assembly in water to form protein transport materials,and the fluorescent labeled BSA protein was combined with electrostatic interaction to carry out in vitro endocytosis experiments.Inverted fluorescence microscopy,it was demonstrated that the protein delivery material could target transport proteins into cells.In this study,we prepared a protein delivery material consisting mainly of ?-cyclodextrin as the main body,and encapsulated the target,positive and negative electrons by the host-guest interaction.It has a good application prospect in protein delivery system.In fact,this study has a certain degree of versatility,changes in the different targets,and the cross-linking agent on the different stimulus response to the functional group,the protein delivery material can transport protein targeting specific cells,and release the protein.Improve the therapeutic effect,reduce the side effects,in the protein,delivery system has a wide range of applications.