Comparative Metabolite Profiling and Identification of a GalNAc-Conjugated siRNA, siRNA01, in Plasma Prepared with Various Anticoagulants, Serum, and In Vivo Plasma Using LC-UV-HRMS

This research uses minipigs, chosen for their skin’s similarity to human skin, as a focus of our study. This poster evaluates the bead milling and traditional homogenization methods for processing pig skin tissues, with  an emphasis on efficiency and cross-contamination prevention. We highlight the importance of accurate skin layer separation and uniform, high-quality homogenization samples for successful biological analysis. The detailed process of isolating various skin layers underscores the crucial role of homogenization in ensuring accurate results.Hepatocytes are recommended as a good in vitro metabolism model for oligonucleotides because they closely mimic the uptake into specific compartments of hepatocytes in vivo. However, there are some challenges in using hepatocytes to study the metabolism of GalNAc-conjugated siRNAs compared to that of conventional small molecules. In this study, we selected siRNA01, a commercially available GalNAc-conjugated siRNA, as a model compound for a comprehensive investigation into both in vitro and in vivo metabolism. A detailed comparison of in vivo and in vitro metabolite profiles in rats was conducted.