Main description:

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.

Contents:

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 Discovery

Wing W. Lam, Jie Chen, Rongfang Fran Xu, Jose Silva, and Heng-Keang Lim

27. Drug, Lipid, and Acylcarnitine Profiling Using Dried Blood Spot (DBS) Technology in Drug Discovery

Wensheng Lang, Jenson Qi, and Gary W. Caldwell

28. In Vitro Trapping and Screening of Reactive Metabolites Using Liquid Chromatography-Mass Spectrometry

Zhengyin Yan and Gary W. Caldwell

29. Quantitative Assessment of Reactive Metabolites

Jie Chen, Rongfang Fran Xu, Wing W. Lam, Jose Silva, and Heng Keang Lim

30. In Vitro Assessment of the Reactivity of Acyl Glucuronides

Rongfang Fran Xu, Wing W. Lam, Jie Chen, Michael McMillian, Jose Silva, and Heng Keang Lim

31. In Vitro COMET Assay for Testing Genotoxicity of Chemicals

Haixia Lin, Nan Mei, and Mugimane G. Manjanatha

32. Assessing DNA Damage Using a Reporter Gene System

Michael Biss and Wei Xiao

33. Improved AMES Test for Genotoxicity Assessment of Drugs: Preincubation Assay Using a Low Concentration of Dimethyl Sulfoxide

Atsushi Hakura

34. Methods for Using the Mouse Lymphoma Assay to Screen for Chemical Mutagenicity and Photo-Mutagenicity

Nan Mei, Xiaoqing Guo, and Martha M. Moore