Metabolite Profiling and Identification of Oligonucleotide in In Vitro Metabolic System

Oligonucleotides are synthetic single or double strands of modified RNA or DNA, they are less than 100 nucleotides (Nt) and can be used to modulate gene expression by binding to RNA or DNA through the Watson-Crick base pairing principle. In theory, oligonucleotides can target any gene of interest because only the nucleotide sequence of interest along the targeted DNA or RNA needs to be selected. This characteristic makes oligonucleotides an exciting therapeutic option within precision medicine.

Based on the Ion-pairing LC-UV-high resolution mass spectrometry (Ion-pairing LC-UV-HRMS) technology, a serials in vitro metabolism systems were thoroughly investigated and optimized. The results of the internal validation studies exhibit high correlation of liver targeted siRNA metabolites between in in vitro liver S9 fractions and liver homogenate. Additionally, the major metabolites detected in both in vitro systems were consistent with those observed in in vivo liver performed in internal lab, as well as reported research paper. To thoroughly characterize the relevant metabolites for oligonucleotides in in vitro metabolism, a comprehensive strategy was established with the aim of facilitating the evaluation of metabolism, particularly for liver-targeted oligonucleotides.