title: A Low Protein Binding Cationic Poly(2-oxazoline) as Non-Viral Vector.
abstract: Developing safe and efficient non-viral gene delivery systems remains a major challenge. We present a new cationic poly(2-oxazoline) (CPOx) block copolymer for gene therapy that was synthesized by sequential polymerization of non-ionic 2-methyl-2-oxazoline and a new 2-oxazoline monomer, 2-(N-methyl, N-Boc-amino)-methyl-2-oxazoline, followed by deprotection of the pendant secondary amine groups. Upon mixing with plasmid DNA (pDNA), CPOx forms small (diameter ≈80 nm) and narrowly dispersed polyplexes (PDI <0.2), which are stable upon dilution in saline and against thermal challenge. These polyplexes exhibited low plasma protein binding and very low cytotoxicity in vitro compared to the polyplexes of pDNA and poly(ethylene glycol)-b-poly(L-lysine) (PEG-b-PLL). CPOx/pDNA polyplexes at N/P = 5 bound considerably less plasma protein compared to polyplexes of PEG-b-PLL at the same N/P ratio. This is a unique aspect of the developed polyplexes emphasizing their potential for systemic delivery in vivo. The transfection efficiency of the polyplexes in B16 murine melanoma cells was low after 4 h, but increased significantly for 10 h exposure time, indicative of slow internalization of polyplexes. Addition of Pluronic P85 boosted the transfection using CPOx/pDNA polyplexes considerably. The low protein binding of CPOx/pDNA polyplexes is particularly interesting for the future development of targeted gene delivery.
authors.affiliation: Center for Nanotechnology in Drug Delivery, Division of Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, NC 27599, USA.
authors.forename: Zhijian
authors.initials: Z
authors.lastname: He
authors.affiliation: Center for Nanotechnology in Drug Delivery, Division of Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, NC 27599, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Miao
authors.affiliation: Department Chemie, Technische Universität Dresden, Mommsenstr. 4, 01069 Dresden, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Jordan
authors.affiliation: Center for Nanotechnology in Drug Delivery, Division of Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, NC 27599, USA.
authors.forename: Devika
authors.initials: D
authors.lastname: S-Manickam
authors.affiliation: Functional Polymer Materials, Chair for Chemical Technology of Materials Synthesis, Universität Würzburg, 97070 Würzburg, Germany. robert.luxenhofer@uni-wuerzburg.de.
authors.forename: Robert
authors.initials: R
authors.lastname: Luxenhofer
authors.affiliation: Laboratory for Chemical Design of Bionanomaterials, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow 119899, Russia. kabanov@email.unc.edu.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Kabanov
mesh_terms: D000818:Animals; D018014:Gene Transfer Techniques; D008545:Melanoma / Q000235:genetics*; D051379:Mice; D010080:Oxazoles / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry*; D011060:Poloxalene; D011485:Protein Binding; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D010080:Oxazoles; C422648:pluronic block copolymer p85; C577913:poly(2-oxazoline); D011060:Poloxalene
keywords: biocompatibility; click chemistry; gene delivery; macrophage transfection; non-fouling
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