Zhengyin Yan, Caldwell G.W. (eds.) Cytochrome P450: In Vitro Methods and Protocols

Humana, 2021. — 376 p.This collection explores detailed experimental protocols necessary for setting up a variety of in vitro cytochrome P450 (CYP) assays that are vital in selecting drug candidates in a drug discovery pipeline. Major factors affecting drug metabolism include CYP expression levels, kinetic parameters for individual CYP enzymes, CYP inhibition and induction, time-dependent inhibition (TDI), CYP stability, non-CYP stability, UDP-glucuronosyltransferases (UGT) stability, excretion mechanisms, and drug-drug interactions (DDI), all addressed in this volume. Written for the Methods in Pharmacology and Toxicology series, chapters include helpful background information on the in vitro assay, a list of all the materials, reagents, and equipment necessary to carry out the assay, a step-by-step protocol, notes containing common and unexpected experimental problems in the assay, as well as references containing important supplementary reading.
Authoritative and practical, Cytochrome P450: In Vitro Methods and Protocols serves as a key guide for researchers in the area of discovery and development of new medicines.Preface
Contributors
Cytochrome P450: In Vitro Methods and Protocols
Characteristics of Major Drug Metabolizing Cytochrome P450 Enzymes
Quantitative Determination of Cytochrome P450 Using LC-MS/MS
Spectral Quantitation of Total Cytochrome P450 in Microsomes Using a Single-Beam Spectrophotometer
Cytochrome P450 Enzyme Kinetics: Km and Vmax Determinations Using Metabolite Formation and Substrate Depletion Meth...
Cytochrome P450 Inhibition Assay Using Human Liver Microsomes
Evaluation of Time-Dependent Cytochrome P450 Inhibition Using the Area Under the Curve Shift Method
Assessing Cytochrome P450 Time-Dependent Inhibition (IC50 Shift Assay) Using Both Diluted and Non-Diluted Methods
Cytochrome P450 Gene Regulation: Reporter Assays to Assess Pregnane X Receptor (PXR, NR1I2) Activation
Cytochrome P450 Gene Regulation: Reporter Assays to Assess Aryl Hydrocarbon Receptor (HLHE76, AhR) Activation and...
Cytochrome P450 Gene Regulation: Reporter Assays to Assess Direct and Indirect Activation of Constitutive Androsta...
Determination of In Vitro Cytochrome P450 Induction Potential Using Cryopreserved Human Hepatocytes
Assessing Cytochrome P450 3A (CYP3A) Induction and Suppression Using Primary Human Hepatocyte Spheroids
Cytochrome P450-Mediated Metabolic Stability Assay in Liver Microsomes
Assessment of Cytochrome P450 Metabolic Clearance Using Hepatocyte Suspension
Cytochrome P450 Knock-Out Assay with Nonselective Inhibitors
Differentiation of Cytochrome P450-Mediated from Non-CYP-Mediated Metabolism: Aldehyde Oxidase and Xanthine Oxidor...
In Vitro Reaction Phenotyping of Cytochrome P450 Enzymes
Cytochrome P450-Mediated Drug Bioactivation Assay: An Untargeted High Resolution Accurate Mass LC/MS Assay
Simultaneously Assessing Concentration Changes in 20 Biochemical Pathways as a Result of Drug Dosing and Cytochrom...
Simultaneously Assessing Concentration Changes in 17 Biochemical Pathways as a Result of Drug Dosing and Cytochrom...
Assessing Amino Acid Concentration Changes as a Result of Drug Dosing and Cytochrome P450 and Non-cytochrome P450-...
Index