Simultaneously Assessing Concentration Changes in 20 Biochemical Pathways as a Result of Drug Dosing and Cytochrome P450 and Non-cytochrome P450-Mediated Metabolism: An Untargeted Metabolomics GC/MS Assay

When a drug is dosed into a hepatic cell culture, an animal, or a human, dozens of intracellular metabolites that are structurally related to the drug molecule are produced from a number of cellular metabolism enzymes, including cytochrome P450 (CYP) and non-CYP enzymes. The drug or its metabolites potentially may cause adverse effects on the structure and/or functional properties of one or more cell components, and thus resulting in changes to normal physiology and possibly inducing toxicity. Gas chromatography mass spectrometry (GC/MS) enables the detection and characterization of thousands of small metabolites, and, thus, can be utilized for metabolomics biochemical pathway studies. A GC/MS untargeted metabolomics assay, utilizing an Agilent 7890A/5975 MSD system, is described that can detect metabolite changes in over 20 endogenous biochemical pathways, and thus may be used in conjunction with typical Drug Metabolism and Pharmacokinetics (DMPK) assays to detect off-target and toxicity issues that might arise due to drug/metabolite exposure. Detection of drug/metabolite-induced side effects in preclinical assays will improve the go/no-go selection criteria and drug-design optimization strategies to advance small molecule drug discovery.