Main description:

Recent analyses of drug attrition rates reveal that a significant number of drug candidates fail in the later stage of clinical development owing to absorption, distribution, metabolism, elimination (ADME), and toxicity issues. Lead optimization in drug discovery, a process attempting to uncover and correct these defects of drug candidates, is highly beneficial in lowering the cost and time to develop therapeutic drugs by reducing drug candidate failures in development. At present, parallel synthesis combining with high-throughput screening has made it easier to generate highly potent compounds (i. e. , hits). However, to be a potential drug, a hit must have drug-like characteristics in addition to potency, which include optimal physicochemical properties, reasonable ph- macokinetic parameters, and good safety profiles. Therefore, research tools must be available in drug discovery to rapidly screen for compounds with favorable drug-like properties, and thus adequate resources can be directed to projects with high potential. Optimization in Drug Discovery: In Vitro Methods is a compilation of detailed experimental protocols necessary for setting up a variety of assays important in compound evaluation. A total of 25 chapters, contributed by many experts in their research areas, cover a wide spectrum of subjects including physicochemical properties, abso- tion, plasma binding, metabolism, drug interactions, and toxicity. A good pharmacokinetic profile has long been recognized as an imp- tant drug-like characteristic. Pharmacokinetic parameters are affected by many properties of drug molecules such as physicochemical nature, abso- tion, metabolic stability, and so on.

Contents:

pKa, Solubility and Lipophilicity: Assessing Physicochemical Properties of Lead Compounds
Yushen Guo and Hong Shen

Use of Caco-2 Cell Monolayers to Study Drug Absorption and Metabolism
Ming Hu, Jie Ling, Huimin Lin, and Jun Chen

Absorption Screening Using the PAMPA Approach
Jeffrey A. Ruell and Alex Avdeef

In Situ Single-Pass Perfused Rat Intestinal Model for Absorption and Metabolism
Eun Ju Jeong, Yan Liu, Huimin Lin, and Ming Hu

In Vitro Permeation Study With Bovine Brain Microvessel Endothelial Cells
Seong-Hee Park, Sung-Hack Lee, Yaming Su, and Patrick J. Sinko

An Enzymatic Microplate Assay for Testing P-Glycoprotein Substrates and Inhibitors
S. Orlowski, J. Nugier, and Eric Ezan

Evaluation of Drug-Transporter Interactions Using In Vitro Cell Models
Yaming Su and Patrick J. Sinko

Plasma Protein-Binding Methods in Drug Discovery
Lucinda H. Cohen

Isothermal Titration Calorimetry Characterization of Drug-Binding Energetics to Blood Proteins
Gary W. Caldwell and Zhengyin Yan

Metabolic Stability Assessed by Liver Microsomes and Hepatocytes
David C. Ackley, Kevin T. Rockich, and Timothy R. Baker

In Vitro Drug Metabolite Profiling Using Hepatic S9 and Human Liver Microsomes
Wu-Nan Wu and Linda A. McKown

In Vitro Identification of UDP-glucuronosyltransferases Involved in Drug Metabolism
Michael H. Court

In Vitro CYP Induction in Human Hepatocytes
Daniel R. Mudra and Andrew Parkinson

High-Throughput Screening of Human Cytochrome P450 Inhibitors Using Fluorometric Substrates: Methodology for 25 Enzyme/Substrate Pairs
David M. Stresser

Evaluation of Cytochrome P450 Inhibition in Human Liver Microsomes
Zhengyin Yan and Gary W. Caldwell

Identification of CYP Mechanism-Based Inhibitors
Amin A. Nomeir, Jairam Palamanda, and Leonard Favreau

Detection of DNA Adducts by 32P-Postlabeling Analysis
Naomi Suzuki, Padmaja M. Prabhu, and Shinya Shibutani

Covalent DNA Adduct Formation Mediated by Cytochrome P450
Marie Stiborova

Application of In Vitro Comet Assay for Genotoxicity Testing
Bojana Zegura and Metka Filipic

Assessing DNA Damage Using a Reporter Gene System
Xuming Jia and Wei Xiao

Improvement of the Ames Test Using Human Liver S9 Preparation
Atsushi Hakura, Satoshi Suzuki, and Tetsuo Satoh

Screening for Chemical Mutagens Using the Mouse Lymphoma Assay
Tao Chen and Martha M. Moore

A High-Throughput Binding Assay for HERG
Keith Finlayson and John Sharkey

In Vitro Drug Metabolism: Thiol Conjugation
Wei Tang and Randy R. Miller

In Vitro Screening Assay of the Reactivity of Acyl Glucuronides
Sebastien Bolze

Index