Cationic lipids bearing succinic-based, acyclic and macrocyclic hydrophobic domains: Synthetic studies and in vitro gene transfer.
In this communication we describe the construction of four succinic-based cationic lipids, their formulation with plasmid DNA (pDNA), and an evaluation of their in vitro gene delivery into Chinese hamster ovarian (CHO-K1) cells. The cationic lipids employed in this work possess either a dimethylamine or trimethylamine headgroup, and a macrocyclic or an acyclic hydrophobic domain composed of, or derived from two 16-atom, succinic-based acyl chains. The synthesized lipids and a co-lipid of neutral charge, either cholesterol or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), were formulated in an overall 3:2 cationic-to-neutral lipid molar ratio, then complexed with plasmid DNA (pDNA). The relative transfection performance was evaluated via a comparison between matched versus mismatched formulations defined by the rigidity relationship between the lipids employed. Gel electrophoresis was used to characterize the binding of the lipid formulations with plasmid DNA and the relative degree of plasmid degradation using a DNase I degradation assay. Small angle X-ray diffraction (SAXD) was employed to characterize the packing morphology of the lipid-DNA complexes. In general, the succinic unit embedded within the hydrophobic domain of the cationic lipids was found to improve lipid hydration. The transfection assays revealed a general trend in which mismatched formulations that employed a rigid lipid combined with a non-rigid (or flexible) lipid, outperformed the matched formulations. The results from this work suggest that the design of the cationic lipid structure and the composition of the lipoplex formulation play key roles in governing the transfection performance of nonviral gene delivery agents.
Université; Paris-Sud, EA 401, IFR 141, Faculté de pharmacie, F-92296, Châtenay Malabry, France.
Emile
E
Jubeli
School of Chemistry and Chemical Engineering, Queen's University Belfast, David Keir Building, Stranmillis Road, Belfast, Northern Ireland, BT9 5AG, United Kingdom.
Amanda B
AB
Maginty
Research Division, Weill Cornell Medicine in Qatar, Education City, P.O. Box 24144, Doha, Qatar.
Nada Abdul
NA
Khalique
Research Division, Weill Cornell Medicine in Qatar, Education City, P.O. Box 24144, Doha, Qatar.
Liji
L
Raju
Department of Chemistry, Norwegian University of Science and Technology (NTNU), 7491 Trondheim, Norway.
David G
DG
Nicholson
Department of Physics, University of Stavanger, 4036, Stavanger, Norway.
Helge
H
Larsen
Premedical Unit, Weill Cornell Medicine in Qatar, Education City, P.O. Box 24144, Doha, Qatar. Electronic address: mdp2001@qatar-med.cornell.edu.
Michael D
MD
Pungente
School of Chemistry and Chemical Engineering, Queen's University Belfast, David Keir Building, Stranmillis Road, Belfast, Northern Ireland, BT9 5AG, United Kingdom. Electronic address: goldring@ruc.dk.
William P D
WP
Goldring
D000818:Animals; D016466:CHO Cells; D002412:Cations / Q000737:chemistry; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D018014:Gene Transfer Techniques / Q000592:standards; D006839:Hydrocarbons, Acyclic; D057927:Hydrophobic and Hydrophilic Interactions; D008055:Lipids / Q000737:chemistry*; D047028:Macrocyclic Compounds; D010957:Plasmids; D013386:Succinates / Q000737:chemistry* / Q000378:metabolism; D014162:Transfection / Q000379:methods* / Q000592:standards
D002412:Cations; D006839:Hydrocarbons, Acyclic; D008055:Lipids; D047028:Macrocyclic Compounds; D013386:Succinates; D004247:DNA
Cationic; In vitro DNA transfer; Lipoplex formulation; Macrocyclic lipids; Succinic