Bioanalysis and Biotransformation of Oligonucleotide Therapeutics by Liquid Chromatography‐mass Spectrometry

Abstract

Since 2016, eight new oligonucleotide therapies have been approved which has led to increased interest in oligonucleotide analysis. There is a particular need for powerful bioanalytical tools to study the metabolism and biotransformation of these molecules. This review provides the background on the biological basis of these molecules as currently used in therapies. The article also reviews the current state of analytical methodology including state of the art sample preparation techniques, liquid chromatography‐mass spectrometry methods, and the current limits of detection/quantitation. Finally, the article summarizes the challenges in oligonucleotide bioanalysis and provides future perspectives for this emerging field. © 2020 John Wiley & Sons Ltd.

Bioanalysis in the Age of New Drug Modalities

“…The advantages of LC-MS-based techniques include high specificity, excellent accuracy and precision with high reproducibility, and wide dynamic ranges. For the quantification of oligonucleotides, a variety of sample preparation approaches such as Trizol extraction (liquid-liquid extraction), proteinase K digestion, solid-phase extraction, immunocapture, or hybridization of these pretreatment strategies have been well studied (21). With the growing popularity of high-resolution mass spectrometry (HRMS), the LC-high-resolution accurate mass (LC-HRAM) assays bring about simultaneous metabolite identification and quantification of oligonucleotide.…”

Regulatory guidelines and preclinical tools to study the biodistribution of RNA therapeutics

“…Indeed, RNA therapeutics in a biological matrix contains high concentrations of endogenous RNA, as well as proteins and other biomolecules that can form strong interactions with RNA. MS-based RNA analysis (or their metabolites) then becomes a challenge of robust and efficient sample preparation and analyte extraction prior to analysis (e.g., through trizol extraction, protein digestion, solid-phase/bead-based extraction, and online trapping columns) [83] .…”

Novel advances in biotransformation and bioactivation research – 2020 year in review

“…Commentary: ASOs mediate the knockdown of specific genes by binding to the complementary sequence of targeted mRNA and, therefore, blocking mRNA translation or inducing degradation by RNase H , Sutton et al 2020). Unmodified…”

“…ASOs mediate the knockdown of specific genes by binding to the complementary sequence of targeted mRNA and, therefore, blocking mRNA translation or inducing degradation by RNase H ( Shen and Corey 2018 ; Sutton et al 2020 ). Unmodified ASOs with phosphodiester (PO) backbones are highly susceptible to nuclease degradation in biological matrices, so that chemical modifications are introduced to improve stability, as well as cellular delivery and target binding ( Figure 9 ).…”