title: Anchoring cationic amphiphiles for nucleotide delivery: significance of DNA release from cationic liposomes for transfection.
abstract: We have designed and synthesized lithocholic acid-based cationic amphiphile molecules as components of cationic liposomes for gene transfection (lipofection). To study the relationship between the molecular structures of those amphiphilic molecules, particularly the extended hydrophobic appendant (anchor) at the 3-hydroxyl group, and transfection efficiency, we synthesized several lithocholic and isolithocholic acid derivatives, and examined their transfection efficiency. We also compared the physico-chemical properties of cationic liposomes prepared from these derivatives. We found that isolithocholic acid derivatives exhibit higher transfection efficiency than the corresponding lithocholic acid derivatives. This result indicates that the orientation and extension of hydrophobic regions influence the gene transfection process. Isolithocholic acid derivatives showed a high ability to encapsulate DNA in a compact liposome-DNA complex and to protect it from enzymatic degradation. Isolithocholic acid derivatives also facilitated the release of DNA from the liposome-DNA complex, which is a crucial step for DNA entry into the nucleus. Our results show that the transfection efficiency is directly influenced by the ability of the liposome complex to release DNA, rather than by the DNA-encapsulating ability. Molecular modeling revealed that isolithocholic acid derivatives take relatively extended conformations, while the lithocholic acid derivatives take folded structures. Thus, the efficiency of release of DNA from cationic liposomes in the cytoplasm, which contributes to high transfection efficiency, appears to be dependent upon the molecular shape of the cationic amphiphiles.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan. hirashim@phar.nagoya-cu.ac.jp
authors.forename: Naohide
authors.initials: N
authors.lastname: Hirashima
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Minatani
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Hattori
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Ohwada
authors.forename: Mamoru
authors.initials: M
authors.lastname: Nakanishi
mesh_terms: D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002412:Cations / Q000737:chemistry*; D002627:Chemistry, Physical / Q000379:methods; D002522:Chlorocebus aethiops; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D017930:Genes, Reporter; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008081:Liposomes; D008095:Lithocholic Acid / Q000138:chemical synthesis / Q000737:chemistry; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D015394:Molecular Structure; D041681:NIH 3T3 Cells; D011073:Polyamines / Q000138:chemical synthesis / Q000737:chemistry; D013501:Surface-Active Agents / Q000138:chemical synthesis / Q000378:metabolism*; D014162:Transfection
chemical_list: D002412:Cations; D008081:Liposomes; D011073:Polyamines; D013501:Surface-Active Agents; D008095:Lithocholic Acid; D004247:DNA; D008156:Luciferases