Delivery of plasmid DNA for gene therapy often provokes an inflammatory response that reduces transgene expression. Cationic lipids for lipofection lack pharmacological activity despite the hydrophobicity of many drug candidates that could be exploited. We report a one-step synthesis of a water-soluble, dexamethasone-spermine (DS) cationic lipid that has potent gene transfer capability in confluent endothelial cells when used with the neutral lipid, dioleoylphosphatidylethanolamine (DOPE). In contrast, unconjugated mixtures of dexamethasone, spermine, and/or DOPE have essentially no gene transfer activity. DS retains partial corticosteroid character as quantified by the rapid translocation of glucocorticoid receptor to the nucleus and by dose-dependent transactivation from a glucocorticoid response element. DS has anti-inflammatory activity in vivo in the mouse thioglycollate model of inflammation. In a mouse lung model, DS:DOPE resulted in significantly less interferon-gamma production at Day 1 and elevated transgene expression at Days 1 and 7 postintranasal instillation compared to DC-Chol:DOPE (sterol:DOPE:phosphate molar ratio of 1:1:1). Cationic pharmacophores such as DS represent a new approach to gene delivery and localized therapy.