Preparation Of Tumor Targeting ADM/FA-LNT-OA Nano Drug Delivery System And Its Molecular Mechanism

In an effort to develop a natural polysaccharide targeted nanomicelles drug delivery system loaded with the broad-spectrum anti-tumor drug adriamycin(ADM),FA-LNT-OA polymer was prepared by esterification and acylation reactions,the ADM/FA-LNT-OA polymer nanomicelles drug delivery system was prepared by the dialysis method and optimized by response surface methodology.The structure,morphology,and critical micelle concentration(CMC)of polymer nanomicelles were studied by Fourier infrared spectroscopy,nuclear magnetic resonance spectroscopy,scanning electron microscope,transmission electron microscope,and fluorescence probe.The biocompatibility,pharmacodynamics,and pharmacokinetics of ADM/FA-LNT-OA polymer nanomicelles were investigated by hemolysis,cytotoxicity,in vitro release,stability,molecular docking,in vitro anti-tumor cell proliferation,and high-performance liquid chromatography.The results showed that the optimal conditions for the preparation of polymer nanomicelles ADM/FA-LNT-OA were: the drug loading ratio was ADM:FA-LNT-OA=3:5,the stirring speed was 610 r/min,and the reaction time was 3 h.Under these conditions,the drug loading rate was 14.36±0.12%,and the encapsulation rate was95.17±0.13%.The morphological observation demonstrated that the drug-loaded micelles were uniformly distributed spheres,with a particle size of 180.26±2.44 nm,and the CMC was 0.048 mg/m L as calculated by the pyrene fluorescence probe assay.The hemolysis test and MTT cytotoxicity assay indicated that FA-LNT-OA had good bio-compatibility.The in vitro release results showed that the release of ADM/FA-LNT-OA polymer nanomicelles was sensitive to p H.Stability tests indicated that the particle size,drug release amount and ADM content did not change significantly when the drug-loaded micelles were stored at 0±2℃ and 60±5% humidity for a long time in a light-proof environment,which showed the stability was good.The results of molecular dynamics studies indicated that the ADM/FA-LNT-OA polymeric drug-loaded nanomicelles bound well to the target proteins through hydrogen bonding and hydrophobic interactions.In vitro anti-tumor pharmacodynamic studies proved that the half-inhibitory concentration of ADM/FA-LNT-OA on He La,ES-2 and A549 cells were 48.49±0.25,63.56±0.68 and 105.93±17.03 μg/m L,respectively.Moreover,the pharmacokinetic assays showed that the plasma elimination half-life of ADM/FA-LNT-OA nanomicelles was5.34±0.96 h,the peak time was 0.08±0.03 h,the plasma clearance was 0.02±0.007 L/h/kg,and the maximum plasma concentration was 48.22±1.09 mg/L.Compared with the common ADM formulation,the ADM/FA-LNT-OA nanomicelles prolonged the clearance half-life of ADM,decreased the clearance rate,and extended the retention time of ADM in vivo,which paved the way for an enhancement in the medicinal benefits and a decrease in the therapeutic toxic and adverse effects of ADM.