LC/MS APPLICATIONS IN DRUG DEVELOPMENT Wiley-Interscience Series on Mass Spectrometry Series Editors Dominic M. Desiderio Departments of Neurology and Biochemistry University of Tennessee Health Science Center Nico M. M. Nibbering University of Amsterdam The aim of the series is to provide books written by experts in the various disciplines of mass spectrometry, including but not limited to basic and fundamental research, instrument and methodological developments, and applied research. Books in the Series Michael Kinter, Protein Sequencing and Identification Using Tandem Mass Spectrometry 0-471-32249-0 Mike S. Lee, LC/MS Applications in Drug Development 0-471-40520-5 Forthcoming Books in the Series Chhabil Dass, Principles and Practice of Biological Mass Spectrometry 0-471-33053-1 LC/MS APPLICATIONS IN DRUG DEVELOPMENT Mike S. Lee A JOHN WILEY & SONS, INC., PUBLICATION This book is printed on acid-free paper. • ᭺ Copyright © 2002 by John Wiley & Sons, Inc., New York. 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Library of Congress Cataloging-in-Publication Data: Library of Congress Cataloging-in-Publication Data is available. 0-471-40520-5 Printed in the United States of America. 10987654321 CONTENTS Preface ix Acknowledgments xi 1. Introduction 1 Emerging Analytical Needs / 1 Integration of LC/MS into Drug Development / 3 Partnerships and Acceptance / 6 Overview / 10 2. Drug Development Overview 11 Analysis Perspectives / 11 The Four Stages of Drug Development / 12 Drug Discovery / 14 Preclinical Development / 15 Clinical Development / 16 Manufacturing / 17 3. Accelerated Drug Development 19 Accelerated Development Strategies / 20 Quantitative and Qualitative Process Elements / 20 v Quantitative Process Pipeline / 24 Qualitative Process Pipeline / 25 Motivating Factors / 27 Analysis Opportunities for Accelerated Development / 28 Full-Time Equivalent / 28 Sample Throughput Model / 29 Elimination Model / 29 Rate-Determining Event Model / 31 Accelerated Development Perspectives / 33 4. LC/MS Development 34 The Elements of LC/MS Application / 34 HPLC / 35 Mass Spectrometry / 35 LC/MS Interface / 36 LC/MS Growth / 38 5. Strategies 41 Standard Methods / 43 Template Structure Identification / 46 Databases / 49 Screening / 50 Integration / 53 Miniaturization / 55 Parallel Processing / 56 Visualization / 58 Automation / 61 Summary / 63 6. LC/MS Applications 65 Drug Discovery / 65 Proteomics / 68 Protein Expression Profiling / 70 Quantitation / 76 vi CONTENTS Glycoprotein Mapping / 78 Natural Products Dereplication / 83 Lead Identification Screening / 88 Bioaffinity Screening / 89 Combinatorial Library Screening / 92 Open-Access LC/MS / 96 Structure Confirmation / 97 High Throughput / 100 Purification / 102 Combinatorial Mixture Screening / 103 In Vivo Drug Screening / 106 Pharmacokinetics / 109 In Vitro Drug Screening / 115 Metabolic Stability Screening / 118 Membrane Permeability / 119 Drug-Drug Interaction / 121 Metabolite Identification / 122 Preclinical Development / 123 Metabolite Identification / 125 Impurity Identification / 132 Degradant Identification / 140 Clinical Development / 145 Quantitative Bioanalysis—Selected Ion Monitoring / 148 Quantitative Bioanalysis—Selected Reaction Monitoring / 152 Quantitative Bioanalysis—Automated Solid-Phase Extraction / 156 Quantitative Bioanalysis—Automated On-Line Extraction / 162 Metabolite Identification / 165 Degradant Identification / 168 Manufacturing / 171 Impurity Identification Using Data-Dependent Analysis / 173 CONTENTS vii Peptide Mapping in Quality Control / 176 Patent Protection / 178 7. Future Applications and Prospects 183 Workstations / 183 Multidimensional Analysis / 186 Miniaturization / 187 Information Management / 189 Strategic Outsourcing / 190 Summary / 191 8. Perspectives on the Future Growth of LC/MS 192 9. Conclusions 195 Glossary 197 References 205 Index 235 viii CONTENTS PREFACE The combination of high-performance liquid chromatography and mass spectrometry (LC/MS) has had a significant impact on drug development over the past decade. Continual improvements in LC/MS interface technologies combined with powerful features for structure analysis, qualitative and quantitative, has resulted in a widened scope of application. These improvements coincided with breakthroughs in combinatorial chemistry, molecular biology, and an overall industry trend of accelerated drug development. The inte- gration of new technologies in the pharmaceutical industry created a situation where the rate of sample generation far exceeds the rate of sample analysis. As a result, new paradigms for the analysis of drugs and related substances have been developed. Both pharma- ceutical and instrument manufacturing industries have mutually benefited. The growth in LC/MS applications has been extensive, with re- tention time and molecular weight emerging as essential analytical features from drug target to product. LC/MS-based methodologies that involve automation, predictive or surrogate models, and open- access systems have become a permanent fixture in the drug devel- opment landscape. An iterative cycle of “what is it?” and “how much is there?” continues to fuel the tremendous growth of LC/MS in the pharmaceutical industry. During this time, LC/MS has become widely accepted as an integral part of the drug development process. ix It is clear that significant developments are happening in the analytical sciences and that future innovations will continue to posi- tively impact the ability for industry scientists to create, share, and collaborate. This book, based on an earlier review (Lee and Kerns, 1999), describes the utility of LC/MS techniques for accelerated drug de- velopment and provides perspective on the significant changes in strategies for pharmaceutical analysis. Specific examples of LC/MS innovation and application highlight the interrelation between the drug development activities that generate samples and the activities responsible for analysis. It should be noted that the extent of LC/MS applications within drug development is hardly complete, and there- fore, this book is not intended to be encyclopedic. The goal was to provide an industry perspective on how and why LC/MS became a premier tool for pharmaceutical analysis. Frequently, the review of a specific methodology or technology creates a barrier of interaction with other disciplines. The applications described in this book are organized with regard to current drug development cycles (i.e., drug discovery, preclinical development, clinical development, manu- facturing) to provide an enabling reference for a wide community of chemists and biologists. Future applications of LC/MS technologies for accelerated drug development and emerging industry trends that deal with sample preparation, chromatography, mass spectrometry, and information management are also discussed. Mike S. Lee x PREFACE [...]... of academic, instrument manufacture, and industry 10 INTRODUCTION interests However, the current industry trends highlight the tremendous challenge of drug development and an expanding need for tools that provide for fast, sensitive, and selective analysis of drugs and drug- related compounds OVERVIEW This book focuses on LC/ MS applications in drug development It examines the role of LC/ MS in the pharmaceutical... accelerated drug development featuring analysis 22 ACCELERATED DRUG DEVELOPMENT Lead Compound Lead Candidate Phase I Drug Discovery NDA/MAA IND/CTA Preclinical Development Phase II Phase III Clinical Development Figure 3.1 Hypothetical drug development pipeline, illustrating activities during a 12-month period Three stages of drug development are indicated at the bottom of the figure The corresponding milestones... of intermediates, drug substance, and drug product, out-of-specification results must be immediately addressed LC/ MS Applications in Drug Development Mike S Lee Copyright  2002 John Wiley & Sons, Inc ISBN: 0-471-40520-5 CHAPTER 3 ACCELERATED DRUG DEVELOPMENT An accelerated drug development strategy focuses on producing drug candidates and accomplishing goals in less time than with traditional development. .. acceptance of LC/ MS in the pharmaceutical industry today The ages of an analytical method are translated into stages of LC/ MS events that lead to its routine use in the pharmaceutical industry (Table 1.1) The various stages represent a continuum for LC/ MS advancement, beginning with basic research performed in universities, followed by the design and manufacture of instruments, and concluding with industry... the drug products are also required Clinical Development The clinical development stage comprises three distinct components or phases (I, II, and III), and culminates in the filing of the NDA/MAA Each phase involves process scale-up, pharmacokinetics, drug delivery, and drug safety activities During phase I clinical development, the compound’s safety and pharmacokinetic profile is defined The determination... the primary focus being on accelerating drug development New tools and strategies for analysis combined with technologies such as biomolecular screening, combinatorial chemistry, and genomics have positioned the pharmaceutical industry to harvest discovery and manufacture development opportunities INTEGRATION OF LC/ MS INTO DRUG DEVELOPMENT Liquid chromatography/mass spectrometry (LC/ MS) -based techniques... Mixture X-ray NMR LC/ NMR MS UV IR HPLC/UV LC/ MS Figure 1.1 Structure analysis matrix that illustrates pharmaceutical analysis preferences for four specific sample types: nontrace/pure; nontrace/ mixture; trace/pure; and trace/mixture (Courtesy of Milestone Development Services, Newtown, Pa., USA.) INTEGRATION OF LC/ MS INTO DRUG DEVELOPMENT 5 within the industry, inspiring creativity and generating new perspectives... and/or initiatives that occurred in the pharmaceutical industry during this time In many instances, an analysis is provided to illustrate the result or development situation if LC/ MS was not used In these cases, the impact (number of samples) and value (cost) on drug development is highlighted independent of the technical features of LC/ MS analysis These unique industry perspectives offer an enabling... tremendously due to the increased use of LC/ MS and other information-rich technologies From strictly an analysis perspective, LC/ MS has demonstrated a 6 INTRODUCTION unique capability for maintaining high quality performance and a rapid turnaround of samples Yet, it is the accurate and efficient processing of information that has been essential for LC/ MS use and acceptance As a result, LC/ MS has developed... performance in the pharmaceutical industry will effect relationships with academia and instrument manufacturers is featured These sections extend the initial thesis of accelerated development to include new analysis bottlenecks and perspectives on analysis issues and industry needs LC/ MS Applications in Drug Development Mike S Lee Copyright  2002 John Wiley & Sons, Inc ISBN: 0-471-40520-5 CHAPTER 2 DRUG DEVELOPMENT . 29 Rate-Determining Event Model / 31 Accelerated Development Perspectives / 33 4. LC/ MS Development 34 The Elements of LC/ MS Application / 34 HPLC / 35 Mass Spectrometry / 35 LC/ MS Interface / 36 LC/ MS. Lee, LC/ MS Applications in Drug Development 0-471-40520-5 Forthcoming Books in the Series Chhabil Dass, Principles and Practice of Biological Mass Spectrometry 0-471-33053-1 LC/ MS APPLICATIONS IN DRUG. Four Stages of Drug Development / 12 Drug Discovery / 14 Preclinical Development / 15 Clinical Development / 16 Manufacturing / 17 3. Accelerated Drug Development 19 Accelerated Development Strategies