Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro.

New amphiphilic 1,4-DHP derivative with remoted cationic moieties at positions 2 and 6 was synthesised to study DNA delivery activity. The results were compared with data obtained for cationic 1,4-DHP derivative , which is known to be the most efficient one among the previously tested 1,4-DHP amphiphiles. We analysed the effects of concentration, complexation media, and complex/cell contact time on the gene delivery effectiveness and cell viability. Transmission electron microscopy data confirms that lipoplexes formed by the compound were quite uniform, vesicular-like structures with sizes of about 50 nm, and lipoplexes produced by compound were of irregular shapes, varied in size in the range of 25⁻80 nm. Additionally, confocal microscopy results revealed that both amphiphiles effectively delivered green fluorescent protein expression plasmid into BHK-21 cells and produced a fluorescent signal with satisfactory efficiency, although compound was more cytotoxic to the BHK-21 cells with an increase of concentration. It can be concluded that optimal conditions for lipoplexes delivery in BHK-21 cells were the serum free media without 0.15 M NaCl, at an N/P ratio of 0.9. Compound showed higher transfection efficiency to transfect BHK-21 and Cos-7 cell lines, when transfecting active proliferating cells. Although was not able to transfect all studied cell lines we propose that it could be cell type specific. The compound showed modest delivery activity in all used cell lines, and higher activity was obtained in the case of H2-35 and B16 cells. The transfection efficiency in cell lines MCF-7, HeLa, and Huh-7 appears to be comparable to the reference compound and minimal in the HepG2 cell line.