1. Small Molecule Formulation Screening Strategies in Drug Discovery
Gary W. Caldwell, Becki Hasting, John Masucci, and Zhengyin Yan
2. Assessment of Drug Plasma Protein Binding in Drug Discovery
Dennis Kalamaridis and Nayan Patel
3. Drug Partition in Red Blood Cells
Dennis Kalamaridis and Karen DiLoreto
4. Permeability Assessment Using 5-Day Cultured Caco-2 Cell Monolayers
Gary W. Caldwell, Chrissa Ferguson, Robyn Buerger, Lovonia Kulp, and Zhengyin Yan
5. In Situ Single Pass Perfused Rat Intestinal Model
Maria Markowska and L. Mark Kao
6. Metabolic Stability Assessed by Liver Microsomes and Hepatocytes
Kevin J. Coe and Tatiana Koudriakova
7. Metabolic Assessment in Alamethicin-Activated Liver Microsomes: Co-Activating CYPs and UGTs
Gary W. Caldwell and Zhengyin Yan
8. Phenotyping UDP-Glucuronosyltransferases (UGTs) Involved in Human Drug Metabolism: An Update
Michael H. Court
9. In Vitro CYP/FMO Reaction Phenotyping
Carlo Sensenhauser
10. Human Pregnane X Receptor (hPXR) Activation Assay in Stable Cell Lines
Judy L. Raucy
11. Characterization of Constitutive Androstane Receptor (CAR) Activation
Caitlin Lynch, Haishan Li, and Hongbing Wang
12. DNA Binding (Gel Retardation Assay) Analysis for Identification of Aryl Hydrocarbon (Ah) Receptor Agonists and Antagonists
Anatoly A. Soshilov and Michael S. Denison
13. Cell-Based Assays for Identification of Aryl Hydrocarbon Receptor (AhR) Activators
Guochun He, Jing Zhao, Jennifer C. Brennan, Alessandra A. Affatato, Bin Zhao, Robert H. Rice, and Michael S. Denison
14. In Vitro CYP Induction Using Human Hepatocytes
Monica Singer, Carlo Sensenhauser, and Shannon Dallas
15. Assessment of CYP3A4 Time-Dependent Inhibition in Plated and Suspended Human Hepatocytes
J. George Zhang and David M. Stresser
16. Evaluation of Time-Dependent CYP3A4 Inhibition Using Human Hepatocytes
Yuan Chen and Adrian J. Fretland
17. Rapidly Distinguishing Reversible and Time-Dependent CYP450 Inhibition Using Human Liver Microsomes, Co-Incubation, and Continuous Fluorometric Kinetic Analyses
Gary W. Caldwell and Zhengyin Yan
18. Identification of Time-Dependent CYP Inhibitors Using Human Liver Microsomes (HLM)
Kevin J. Coe, Judith Skaptason, and Tatiana Koudriakova
19. CYP Time-Dependent Inhibition (TDI) Using an IC50 Shift Assay with Stable Isotopic Labeled Substrate Probes to Facilitate Liquid Chromatography/Mass Spectrometry Analyses
Gary W. Caldwell and Zhengyin Yan
20. Screening for P-Glycoprotein (Pgp) Substrates and Inhibitors
Qing Wang and Tina M. Sauerwald
21. In Vitro Characterization of Intestinal Transporter, Breast Cancer Resistance Protein (BCRP)
Chris Bode and Li-Bin Li
22. In Vitro Characterization of Intestinal and Hepatic Transporters: MRP2
Ravindra Varma Alluri, Peter Ward, Jeevan Kunta, Brian C. Ferslew, Dhiren R. Thakker, and Shannon Dallas
23. In Vitro Characterization of Hepatic Transporters OATP1B1 and OATP1B3
Blair Miezeiewski and Allison McLaughlin
24. In Vitro Characterization of Renal Transporters OAT1, OAT3, and OCT2
Ying Wang and Nicole Behler
25. General Guidelines for Setting Up an In Vitro LC/MS/MS Assay
John A. Masucci and Gary W. Caldwell
26. Metabolite Identification in Drug Dis
Thoroughly revised and updated, Optimization in Drug Discovery: In Vitro Methods, Second Edition presents a wide spectrum of in vitro assays including formulation, plasma binding, absorption and permeability, cytochrome P450 (CYP) and UDP-glucuronosyltransferases (UGT) metabolism, CYP inhibition and induction, drug transporters, drug-drug interactions via assessment of reactive metabolites, genotoxicity, and chemical and photo-mutagenicity assays. Written for the Methods in Pharmacology and Toxicology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Expert authors have developed and utilized these in vitro assays to achieve “drug-like” characteristics in addition to efficacy properties and good safety profiles of drug candidates.
Comprehensive and up-to-date,
Optimization in Drug Discovery: In Vitro Methods, Second Edition aims to guide researchers down the difficult path to successful drug discovery and development.
