title: Phospholipid-detergent conjugates as novel tools for siRNA delivery.
abstract: One of the potential benefits of drug delivery systems in medicine is the creation of nanoparticle-based vectors that deliver a therapeutic cargo in sufficient quantity to a target site to enable a selective effect, width of the therapeutic window depending on the toxicity of the vector and the cargo. In this work, we intended to improve the siRNA delivery efficiency of a new kind of nucleic acid carrier, which is the result of the conjugation of the membrane phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) to the membrane-active species Triton X-100 (TX100). We hypothesized that by improving the biodegradability the cytotoxicity of the conjugate might by reduced, whereas its original transfection potential would be tentatively preserved. DOPC was conjugated to Triton X-100 through spacers displaying various resistance to chemical hydrolysis and enzyme degradation. The results obtained through in vitro siRNA delivery experiments showed that the initial phosphoester bond can be replaced with a phospho(alkyl)enecarbonate group with no loss in the transfection activity, whereas the associated cytotoxicity was significantly decreased, as assessed by metabolic activity and membrane integrity measurements. The toxicity of the conjugates incorporating a phospho(alkyl)enesuccinnate moiety proved even lower but was clearly balanced with a reduction of the siRNA delivery efficiency. Hydrolytic stability and intracellular degradation of the conjugates were investigated by NMR spectroscopy and mass spectrometry. A general trend was that the more readily degraded conjugates were those with the lower toxicity. Otherwise, the phospho(alkyl)enecarbonate conjugates revealed some hemolytic activity, whereas the parent phosphoester did not. The reason why these conjugates behave differently with respect to hemolysis might be a consequence of unusual fusogenic properties and probably reflects the difference in the stability of the conjugates in the intracellular environment.
authors.affiliation: Laboratoire de Conception et Application de Molécules Bioactives, CNRS-Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Pierrat
authors.forename: Gilles
authors.initials: G
authors.lastname: Laverny
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Creusat
authors.forename: Patrick
authors.initials: P
authors.lastname: Wehrung
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Strub
authors.forename: Alain
authors.initials: A
authors.lastname: VanDorsselaer
authors.forename: Françoise
authors.initials: F
authors.lastname: Pons
authors.forename: Guy
authors.initials: G
authors.lastname: Zuber
authors.forename: Luc
authors.initials: L
authors.lastname: Lebeau
mesh_terms: D045744:Cell Line, Tumor; D003902:Detergents / Q000737:chemistry* / Q000494:pharmacology*; D016503:Drug Delivery Systems; D005997:Glycerylphosphorylcholine / Q000031:analogs & derivatives* / Q000737:chemistry; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D053758:Nanoparticles / Q000737:chemistry*; D009696:Nucleic Acids / Q000737:chemistry*; D010713:Phosphatidylcholines; D010743:Phospholipids / Q000494:pharmacology*; D010767:Phosphorylcholine / Q000737:chemistry* / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D014162:Transfection
chemical_list: D003902:Detergents; D009696:Nucleic Acids; D010713:Phosphatidylcholines; D010743:Phospholipids; D034741:RNA, Small Interfering; D010767:Phosphorylcholine; D005997:Glycerylphosphorylcholine; C017251:1,2-oleoylphosphatidylcholine