Construction Of Dual-function Pab-fms Immune Mixed Micelle And Anti-A549 Cells Effect In Vitro

Objective: The amphiphilic polymer mPEG-DSPE/mPEG-PLA was used as a carrier material to prepare mixed micelles for carrying CDK inhibitor Palbociclib.The anti M-CSFR monoclonal antibody was coupled to the surface to construct an immune mixed micelle,and its biocompatibility and in vitro antitumor activity were initially explored.Methods: 1.Preparation and characterization of mixed micelles: using film hydration method and single factor experiment to optimize the preparation process to obtain mixed micelles with uniform particle size distribution and high drug loading.Use laser particle size analyzer to detect its particle size distribution,Zeta potential and stability.Ultraviolet spectrophotometric determination of drug loading and in vitro release of mixed micelles.Pyrene fluorescence probe method for detecting CMC.2.Preparation and characterization of immune mixed micelles: anti M-CSFR monoclonal antibody was thiolated and then coupled to the surface of mixed micelles to construct immune mixed micelles,and its particle size distribution and Zeta potential were characterized by laser particle size analyzer.Determination of drug loading of immune mixed micelles by ultraviolet spectrophotometry.3.Evaluation of biocompatibility of immune mixed micelles: explore the biocompatibility of immune mixed micelles through MTT experiment,hemolysis experiment,BSA adsorption experiment.4.Evaluation of anti-tumor activity of immune mixed micelles in vitro: The human lung cancer cell A549(M-CSFR high expression)was selected as the cell model,and the proliferation inhibition effect of free drug and immune mixed micelle on the proliferation of A549 lung cancer cell was detected by MTT method.Observe the effect of immune mixed micelles on apoptosis of A549 cells by fluorescence microscope.Using flow cytometry to detect the effect of immune mixed micelles on A549 cell apoptosis.The effect of immune mixed micelles on migration of A549 cells was detected by scratch test.Results: A single factor experiment was used to optimize the preparation of the prescription to obtain a mixed micelle with a particle size of about 155 nm,a uniform distribution,a Zeta potential of 89.22 mV,and a drug load of up to 25.39%.After thiolation of anti M-CSFR monoclonal antibody,it was effectively coupled to the surface of mixed micelles and the coupling amount was determined to be about 1.21?g/mg.The prepared immune mixed micelles have a particle size of about 159 nm and are evenly distributed,with a Zeta potential of 78.33 mv and a drug loading of 24%.The biocompatibility experiment proves that the hemolysis rate of different concentrations of immune mixed micelles is less than 3%.Compared with free drugs,immune mixed micelles have low protein adsorption and little effect on the proliferation of normal cells.Immune mixed micelles have a strong proliferation inhibitory effect on tumor cells A549,and the inhibitory effect on tumor cells is better than free drugs.In addition,immune mixed micelles can also increase the apoptosis of A549 cells and inhibit their migration.Conclusion: Successfully constructed immune mixed micelles mediated by anti M-CSFR monoclonal antibody,with uniform particle size distribution,stable properties,and good biocompatibility.Compared with free drugs,immune mixed micelles have better antitumor activity.