Date of Completion
metabolic toxicity, high throughput, LCMS, layer-by-layer
Field of Study
Doctor of Philosophy
Metabolic toxicity is one of main concerns in drug discovery and development. High efficient and low cost metabolic toxicity assessment is still a major challenge for pharmaceutical industry. This thesis focuses on the development of high throughput in vitro metabolic toxicity assay coupling with liquid chromatography tandem mass spectrometry for metabolic adducts generation, characterization and quantitation. The key technology was based on electrostatic fabrication of polyelectrolytes and biomolecules on magnetic beads to form bioreactors in 96 well plate format and acquired metabolites induced toxicity molecular information by LC-MS experiments.
Chapter one will address the goals and significance of the metabolic toxicity assay at the early stage of drug candidate studies. It will also provide the essential background of metabolic toxicity, including metabolic enzymes function mechanism, covalent conjugates formation chemistry and consequences, and liquid chromatography mass spectrometry detection methods.
Chapter two will reveal the first discovery on metabolic genetic adducts of benzo[ghi]perylene by the cytochrome P450 enzyme/ DNA assembled biocolloid reactors generation, and followed by LC-MS/MS analysis.
Chapter three will present the high-throughput screening and quantitation assay for comparing genotoxicity profiles towards hepatic and extra-hepatic tissues, which could reveal the metabolic abilities of different organs for specific drugs. The LC-MS result is validated by comet assay.
Chapter four will describe a new approach to generate drug metabolites-protein conjugates and provide damaged site structural information by liquid chromatography mass spectrometry.
Li, Dandan, "Metabolic Toxicity Studies using High Throughput Bioreactor Assay and Liquid Chromatography Coupled with Mass Spectrometry (LC-MS/ MS)" (2014). Doctoral Dissertations. 604.