Novel Cationic Lipids Incorporating an Acid-Sensitive Acylhydrazone Linker:  Synthesis and Transfection Properties

• Published in: Journal of medicinal chemistry Vol. 47; no. 21; pp. 5210 - 5223
• Main Authors: Aissaoui, Abderrahim, Martin, Benjamin, Kan, Erwan, Oudrhiri, Noufissa, Hauchecorne, Michelle, Vigneron, Jean-Pierre, Lehn, Jean-Marie, Lehn, Pierre
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 07.10.2004; Amer Chemical Soc
• Subjects: Animals; Biological and medical sciences; Biotechnology; Cations - chemistry; Cell Line; Chemistry, Medicinal; Cholestenones - chemical synthesis; Cholestenones - chemistry; Cholestenones - toxicity; DNA - administration & dosage; Female; Fundamental and applied biological sciences. Psychology; Gene therapy; Genes, Reporter; Health. Pharmaceutical industry; Humans; Hydrazones - chemical synthesis; Hydrazones - chemistry; Hydrazones - toxicity; Hydrolysis; Industrial applications and implications. Economical aspects; Instillation, Drug; Life Sciences & Biomedicine; Lipids - chemical synthesis; Lipids - chemistry; Lipids - toxicity; Liposomes; Luciferases - biosynthesis; Luciferases - genetics; Lung - metabolism; Medical sciences; Mice; Mice, Inbred BALB C; Miscellaneous; Pharmacology & Pharmacy; Pharmacology. Drug treatments; Science & Technology; Structure-Activity Relationship; Transfection; IN-VITRO; CHOLESTEROL; MECHANISM; DRUG-DELIVERY; DNA; PH; CYTOPLASMIC EXPRESSION; EFFICIENT VECTORS; MEDIATED GENE-TRANSFER; LIPOSOMES; Steroid; Organic cation; Spacer arm; Rodentia; Drug carrier; Guanidines; Guanidinium compounds; In vitro; Biological activity; Genetic transfer; In vivo; Vertebrata; Biological fixation; Mammalia; Transfection; Mouse; Animal; Hydrazone; Chemical synthesis; Gene therapy
• ISSN: 0022-2623; 1520-4804
• DOI: 10.1021/jm0408159

Cationic lipid-mediated gene transfection involves uptake of the lipid/DNA complexes via endocytosis, a cellular pathway characterized by a significant drop in pH. Thus, in the present study, we aimed to explore the impact on transfection efficiency of the inclusion of an acid-sensitive acylhydrazone function in the cationic lipid structure. We synthesized and evaluated the transfection properties of a series of four cationic steroid derivatives characterized by an acylhydrazone linkage connecting a guanidinium-based headgroup to a saturated cholestanone or an unsaturated cholest-4-enone hydrophobic domain. Acid-catalyzed hydrolysis was confirmed for all lipids, its rate being highest for those with a cholestanone moiety. The compound bis-guanidinium bis(2-aminoethyl)amine hydrazone (BGBH)-cholest-4-enone was found to mediate efficient gene transfection into various mammalian cell lines in vitro and into the mouse airways in vivo. In vitro transfection studies with BGBH-cholest-4-enone formulations also showed that incorporation of a degradable acylhydrazone bond led to low cytotoxicity and impacted the intracellular trafficking of the lipoplexes. Thus, our work allowed us to identify a cationic lipid structure with an acid-cleavable acylhydrazone linker capable of mediating efficient gene transfection in vitro and in vivo and it thereby provides a basis for further development of related acid-sensitive gene delivery systems.