Identification of GalNAc-Conjugated Antisense Oligonucleotide Metabolites Using an Untargeted and Generic Approach Based on High Resolution Mass Spectrometry

Analytical Chemistry
Christophe HusserSimone Schadt

Abstract

Antisense oligonucleotides linked by phosphorothioates are an important class of therapeutics under investigation in various pharmaceutical companies. Antisense oligonucleotides may be coupled to high-affinity ligands (triantennary N-acetyl galactosamine = GalNAc) for hepatocyte-specific asialoglycoprotein receptors (ASGPR) to enhance uptake to hepatocytes and to increase potency. Since disposition and biotransformation of GalNAc-conjugated oligonucleotides is different from unconjugated oligonucleotides, appropriate analytical methods are required to identify main cleavage sites and degradation products of GalNAc conjugated and unconjugated oligonucleotides in target cells. A highly sensitive method was developed to identify metabolites of oligonucleotides using capillary flow liquid chromatography with column switching coupled to a high resolution Orbitrap Fusion mass spectrometer. Detection of GalNAc-conjugated oligonucleotides and their metabolites was achieved by combining full scan MS with two parallel MS2 experiments, one data-dependent scan and an untargeted MS2 experiment (all ion fragmentation) applying high collision energy. In the all ion fragmentation scan, a diagnostic fragment originating from the phosphorothioat...Continue Reading

References

Sep 2, 2004·Nature Reviews. Molecular Cell Biology·Hanno Steen, Matthias Mann
Apr 20, 2005·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Guowei DaiGuido Marcucci
Mar 7, 2007·Journal of Pharmaceutical and Biomedical Analysis·Zhongping John LinGuowei Dai
May 13, 2008·Rapid Communications in Mass Spectrometry : RCM·Yan ZouJerome Hochman
Oct 27, 2009·Journal of the American Society for Mass Spectrometry·Teng-yi HuangScott A McLuckey
Mar 11, 2011·Bioanalysis·William D van Dongen, Wilfried M A Niessen
Jun 3, 2011·The Journal of Biological Chemistry·Mingshe ZhuW Griffith Humphreys
Apr 1, 1997·Analytical Chemistry·A ApffelW S Hancock
Oct 24, 2013·Nature Materials·Rosemary KanastyDaniel Anderson
Apr 1, 1995·Journal of the American Society for Mass Spectrometry·M W SenkoF W McLaffertycor
Feb 18, 2015·Advanced Drug Delivery Reviews·Xin Ming, Brian Laing
Jul 15, 2015·Human Gene Therapy·Karin E LundinC I Edvard Smith
Apr 17, 2016·Nucleic Acids Research·Rudolph L Juliano

❮ Previous
Next ❯

Citations

May 31, 2019·Physical Chemistry Chemical Physics : PCCP·Baptiste SchindlerIsabelle Compagnon
Jun 27, 2020·Mass Spectrometry Reviews·James Michael SuttonMichael G Bartlett
Mar 7, 2019·Scientific Reports·Mai B ThayerBrooke M Rock
Sep 16, 2020·Pharmacological Reviews·Ai-Ming YuMei-Juan Tu
Aug 20, 2019·Molecular Therapy. Nucleic Acids·Jaeah KimMichael G Bartlett
Dec 26, 2018·Molecular Therapy. Nucleic Acids·Sabine SewingAnnie Moisan
Feb 4, 2021·Nucleic Acid Therapeutics·William F KiesmanGary Held
Apr 17, 2021·Drug Discovery Today·Lars WeidolfShalini Andersson
Jun 2, 2021·Toxicologic Pathology·Fernando Romero-PalomoAnnamaria Braendli-Baiocco
Apr 30, 2021·Drug Metabolism Reviews·S Cyrus KhojastehCong Wei
Aug 15, 2021·Journal of Pharmaceutical and Biomedical Analysis·Mario ThevisAndreas Thomas
Aug 20, 2020·Journal of the American Society for Mass Spectrometry·J Michael SuttonMichael G Bartlett
Sep 3, 2021·Bioanalysis·Jennifer M NguyenMatthew A Lauber

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antisense Oligonucleotides: ND

This feed focuses on antisense oligonucleotide therapies such as Inotersen, Nusinursen, and Patisiran, in neurodegenerative diseases including amyotrophic lateral sclerosis.