title: Measuring Drug Metabolism Kinetics and Drug-Drug Interactions Using Self-Assembled Monolayers for Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry.
abstract: The competition of two drugs for the same metabolizing enzyme is a common mechanism for drug-drug interactions that can lead to altered kinetics in drug metabolism and altered elimination rates in vivo. With the prevalence of multidrug therapy, there is great potential for serious drug-drug interactions and adverse drug reactions. In an effort to prevent adverse drug reactions, the FDA mandates the evaluation of the potential for metabolic inhibition by every new chemical entity. Conventional methods for assaying drug metabolism (e.g., those based on HPLC) have been established for measuring drug-drug interactions; however, they are low-throughput. Here we describe an approach to measure the catalytic activity of CYP2C9 using the high-throughput technique self-assembled monolayers for matrix-assisted laser desorption-ionization (SAMDI) mass spectrometry. We measured the kinetics of CYP450 metabolism of the substrate, screened a set of drugs for inhibition of CYP2C9 and determined the Ki values for inhibitors. The throughput of this platform may enable drug metabolism and drug-drug interactions to be interrogated at a scale that cannot be achieved with current methods.
authors.affiliation: Department of Pharmacology, Northwestern University Feinberg School of Medicine , Chicago, Illinois 60611, United States.
authors.forename: Lyndsey L
authors.initials: LL
authors.lastname: Anderson
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Berns
authors.forename: Pradeep
authors.initials: P
authors.lastname: Bugga
authors.affiliation: Department of Pharmacology, Northwestern University Feinberg School of Medicine , Chicago, Illinois 60611, United States.
authors.forename: Alfred L
authors.initials: AL
authors.lastname: George
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University Feinberg School of Medicine , Chicago, Illinois 60611, United States.
authors.forename: Milan
authors.initials: M
authors.lastname: Mrksich
mesh_terms: D065729:Cytochrome P-450 CYP2C9 / Q000378:metabolism*; D004347:Drug Interactions; D057166:High-Throughput Screening Assays; D006801:Humans; D007004:Hypoglycemic Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D015394:Molecular Structure; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014044:Tolbutamide / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D007004:Hypoglycemic Agents; D014044:Tolbutamide; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9