Quantification of Biomarkers in Discovery: Probing PK/PD Effect Using Multi-component Analysis and High Resolution Mass Spectrometry

摘要

TCA/methanol yielded the best overall extraction efficiency and chromatographic behavior for both endogenous metabolites and API compounds with different hydrophobicity index (log P). Ultra HR-TOF combined with UPLC was demonstrated to be an effective tool in early drug discovery for obtaining qualitative and quantitative information on both endogenous and API related analytes under chromatographic time scales compatible with typical discovery PK assays. Detection limits of 5 nM were achieved for several model API compounds with a dynamic range of 2.5 to 3 orders of magnitude, with the ability to extend the dynamic range up to 4 orders of magnitude by combining isotopes with different ion intensities. PD modeling with the help of semi-quantitative data of tryptophan and kynurenine together with the in-vivo PK data was successfully employed in API screening in early drug discovery.