Abstract
Mono-antennary galacto derivatives of cholesterol are being actively developed to direct lipoplexes to the asialoglycoprotein receptor (ASGP-R) on hepatocytes. Here we report on a novel ASGP-R ligand cholest-5-en-3-yl [1-(β-d-galactopyranosyl)-1H-1,2,3-triazol-4-yl]methylcarbamate (4), assembled by a copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry), and compare it with cholest-5-en-3-yl-β-d-galactopyranoside (2) and cholest-5-en-3-yl [1-(β-d-galactopyranosyl-1′-oxy)phen-4-yl]carbamate (3), in liposome formulations with or without 5 mol% distearoylphosphatidylethanolamine poly(ethylene glycol)2000, intended for DNA delivery to ASGP-R-positive hepatocyte-derived HepG2 cells and the ASGP-R-negative embryo kidney cell line HEK293. Transfection levels attained with lipoplex 4 were 100 and 300 % greater than those for lipoplexes 2 and 3 respectively in HepG2 cells, while competition assays reduced transfection levels by up to 98 %. Transfection activities achieved in HEK293 cells were up to three orders of magnitude lower. Therefore, 4 is representative of a new class of promising hepatotropic ligands for gene delivery.
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Acknowledgments
This study was supported by the National Research Foundation (NRF), Pretoria, the University of the Witwatersrand (University and Science Faculty Research Councils) and Stellenbosch University (Faculty and Departmental funding), South Africa. RUI thanks the University of the Witwatersrand Research Office and the NRF for a postdoctoral fellowship. The authors also gratefully acknowledge Mr R. Mampa (University of the Witwatersrand) and Mr D. Jagjivan (University of KwaZulu-Natal) for the NMR spectroscopy service. Ms J. Schneider and Ms M. Ismail (Mass Spectroscopy Service, University of Dortmund and Max Planck Institute for Molecular Physiology, Dortmund, Germany) and Dr James Wesley-Smith (Electron Microscopy, Westville campus University of KwaZulu-Natal) are also acknowledged.
Supporting information
Online resource 1: Spectral data.
Online resource 2: Plasmid amplification.
Supplementary Fig. 1 Cryo-TEM micrographs of unPEGylated (a) and PEGylated (b) liposomes and unPEGylated (c) and PEGylated (d) lipoplexes prepared at endpoint ratios.
Supplementary Fig. 2 Cell viability assays on HepG2 and HEK293 cells were conducted under transfection conditions.
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Govender, D., Islam, R.U., De Koning, C.B. et al. Stealth lipoplex decorated with triazole-tethered galactosyl moieties: a strong hepatotropic gene vector. Biotechnol Lett 37, 567–575 (2015). https://doi.org/10.1007/s10529-014-1729-5
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DOI: https://doi.org/10.1007/s10529-014-1729-5