tailieunhanh - Báo cáo khoa học: NovelN,N¢-diacyl-1,3-diaminopropyl-2-carbamoyl bivalent cationic lipids for gene delivery – synthesis,in vitro transfection activity, and physicochemical characterization

NovelN,N¢-diacyl-1,3-diaminopropyl-2-carbamoyl bivalent cationic lipids were synthesized and their physicochemical properties in lamellar assemblies with and without plasmid DNA were evaluated to elucidate the structural requirements of these double-chained pH-sensitive surfactants for potent non-viral gene delivery and expression. | ễFEBS Journal Novel N N -diacyl-1 3-diaminopropyl-2-carbamoyl bivalent cationic lipids for gene delivery - synthesis in vitro transfection activity and physicochemical characterization Michael Spelios and Michalakis Savva Division of PharmaceuticalSciences Arnold Marie Schwartz College of Pharmacy Health Sciences Long Island University Brooklyn NY USA Keywords cationic lipid elasticity FRET gene delivery lipoplex Correspondence M. Savva Division of Pharmaceutical Sciences Arnold Marie Schwartz College of Pharmacy Health Sciences Long Island University 75 DeKalb Avenue Brooklyn NY 11201 USA Fax 1 718 780 4586 Tel 1 718 488 1471 E-mail msavva@ Received 23 September 2007 revised 5 November 2007 accepted 9 November 2007 doi Novel N N -diacyl-1 3-diaminopropyl-2-carbamoyl bivalent cationic lipids were synthesized and their physicochemical properties in lamellar assemblies with and without plasmid DNA were evaluated to elucidate the structural requirements of these double-chained pH-sensitive surfactants for potent non-viral gene delivery and expression. The highest in vitro transfection efficacies were induced at - 4 1 by the dimyristoyl dipalmitoyl and dioleoyl derivatives 1 3lb2 1 3lb3 and 1 3lb5 respectively without inclusion of helper lipids. Transfection activities were reduced in the presence of either 1 2-dioleoyl-sn-glycero-3-phosphoethanolamine alone or in combination with cholesterol for all derivatives except 1 3lb5 which maintained reporter gene expression levels at - 4 1 and yielded increased lipofection activity at a lower charge ratio of - 2 1. Ethidium bromide displacement indicated efficient plasmid DNA binding and compaction by the transfection-competent analogs. Dynamic light-scattering and electrophoretic mobility studies revealed lipoplexes of the active lipids with large particle sizes mean diameter 500 nm and zeta potentials with positive values low ionic strength or below neutrality high ionic strength . .