This study reports the development of a polymeric carrierfor pulmonarysiRNA delivery via inhalation. Poly(α,β-N-(2-hydroxyethyl)-d,l-asparamide) (PHEA) was first functionalized withdivinyl sulfone (DV) to form PHEA-VS, enabling controlled graftingof 1,2-bis(3-aminopropylamino)ethane (bAPAE, of about ∼25 mol%) and poly(2-methyl-2-oxazoline) (PMeOx, of about ∼5 mol %).The resulting PHEA-VS-g-(PMeOx;bAPAE) copolymer containedprotonable amines for efficient siRNA complexation. Potentiometrictitration confirmed strong buffering capacity, while fluorescencestudies indicated pH-responsive membrane interaction, suggesting improvedendosomal escape. Polyplexes formed starting from a polymer/siRNAratio of 5, with diameters below 40 nm, showing stability in mucinsand pulmonary surfactant and protection against RNase degradation.Surface decoration with the cell-penetrating peptide penetratin (Pen)via a terminal alkyne on PMeOx enhanced cellular uptake and siRNArelease. Biocompatibility tests on 16-HBE cells showed viability over80% at high polymer concentrations. Functional assays in MDA-MB-231-LUCcells demonstrated effective gene silencing, particularly at a polymer/siRNAratio of 5. Combined with the favorable aerosolization propertiesof aqueous dispersions, these results highlight PHEA-VS-g-(PMeOx;bAPAE) as a versatile platform for pulmonary siRNA delivery,offering stability, biocompatibility, and targeted intracellular releasefor potential treatment of respiratory diseases.
Read more