Selective blocking of primary amines in branched polyethylenimine with biocompatible ligand alleviates cytotoxicity and augments gene delivery efficacy in mammalian cells

• Pyridoxyl-polyethylenimine (PyP) polymers have been synthesized without altering the total number of amines and the modifying reagent is biocompatible.
• By selectively targeting primary amines, substitution significantly decreased cytotoxicity of bPEI polymer.
• The projected substitution introduced amphiphilicity in the modified polymers.
• One of the formulations, PyP-3, efficiently carried pDNA and siRNA inside the mammalian cells.
• PyP polymers exhibited considerably higher transfection efficiency advocating that the projected carriers can be used for future gene therapy applications.

Recently, polyethylenimines (PEIs) have emerged as efficient vectors for nucleic acids delivery. However, inherent cytotoxicity has limited their in vivo applications. To address this concern as well as to incorporate hydrophobic domains for improving interactions with the lipid bilayers in the cell membranes, we have tethered varying amounts of amphiphilic pyridoxyl moieties onto bPEI to generate a small series of pyridoxyl-PEI (PyP) polymers. Spectroscopic characterization confirms the formation of PyP polymers, which subsequently form stable complexes with pDNA in nanometric range with positive surface charge. The projected modification not only accounts for a decrease in the density of 1° amines but also allows formation of relatively loose complexes with pDNA (cf. bPEI). Alleviation of the cytotoxicity, efficient interaction with cell membranes and easy disassembly of the pDNA complexes have led to the remarkable enhancement in the transfection efficiency of PyP/pDNA complexes in mammalian cells with one of the formulations, PyP-3/pDNA complex, showing transfection in ∼68% cells compared to ∼16% cells by Lipofectamine/pDNA complex. Further, the efficacy of PyP-3 vector has been established by delivering GFP-specific siRNA resulting in ∼88% suppression of the target gene expression. These results demonstrate the efficacy of the projected carriers that can be used in future gene therapy applications.

Figure optionsDownload as PowerPoint slide