Robert A. Copeland

Evaluation of Enzyme Inhibitors in Drug Discovery

A Guide for Medicinal Chemists and Pharmacologists

• Gebundenes Buch

Produktbeschreibung

With enzymes being the most valued and common of drug targets, an understanding of their interactions with inhibitors is critical to successful drug discovery. Now in a second edition, this proven work clearly explains the biochemical data and experimental details underlying the science, arming medicinal chemists and pharmacologists with the tools they need to master the art of applied enzymology for drug discovery. With updated material throughout, two new chapters, and five new appendices, Evaluation of Enzyme Inhibitors in Drug Discovery, Second Edition remains the only book available on the topic.

Produktdetails

• Verlag: Wiley & Sons
• Artikelnr. des Verlages: 1W118488130
• 2. Aufl.
• Seitenzahl: 576
• Erscheinungstermin: 18. März 2013
• Englisch
• Abmessung: 240mm x 161mm x 36mm
• Gewicht: 1035g
• ISBN-13: 9781118488133
• ISBN-10: 111848813X
• Artikelnr.: 36729784

Autorenporträt

ROBERT A. COPELAND, PhD, is Executive Vice President and Chief Scientific Officer at Epizyme, Inc., a biopharmaceutical company in Cambridge, Massachusetts. He is on the Editorial Board of The Journal of Biological Chemistry and a member of the Faculty of 1000. Dr. Copeland has contributed more than 175 publications to the scientific literature and holds eight U.S.-issued patents. He has authored several books in protein science and enzymology, including Enzymes: A Practical Introduction to Structure, Mechanism, and Data Analysis, Second Edition (Wiley).

Inhaltsangabe

FOREWORD TO SECOND EDITION BY CHRISTOPHER T. WALSH xvii PREFACE TO SECOND EDITION xix FOREWORD TO FIRST EDITION BY PAUL S. ANDERSON xxiii PREFACE TO FIRST EDITION xxv ACKNOWLEDGMENTS FROM FIRST EDITION xxix 1. WHY ENZYMES AS DRUG TARGETS? 1 Key Learning Points
1 1.1 Enzymes Are Essential for Life
2 1.2 Enzyme Structure and Catalysis
6 1.3 Permutations of Enzyme Structure During Catalysis
12 1.4 Extension to Other Target Classes
17 1.5 Other Reasons for Studying Enzymes
18 1.6 Summary
21 References
22 2. ENZYME REACTION MECHANISMS 25 Key Learning Points
25 2.1 Initial Binding of Substrate
25 2.2 Noncovalent Forces in Reversible Ligand Binding to Enzymes
28 2.3 Transformations of the Bound Substrate
30 2.4 Steady State Analysis of Enzyme Kinetics
39 2.5 Typical Values of Steady State Kinetic Parameters
46 2.6 Graphical Determination of kcat and KM
47 2.7 Reactions Involving Multiple Substrates
49 2.8 Summary
54 References
54 3. REVERSIBLE MODES OF INHIBITOR INTERACTIONS WITH ENZYMES 57 Key Learning Points
57 3.1 Enzyme-Inhibitor Binding Equilibria
58 3.2 Competitive Inhibition
59 3.3 Noncompetitive Inhibition
68 3.4 Uncompetitive Inhibition
82 3.5 Inhibition Modality in Bisubstrate Reactions
86 3.6 Value of Knowing Inhibitor Modality
88 3.7 Enzyme Reactions on Macromolecular Substrates
96 3.8 Summary
118 References
119 4. ASSAY CONSIDERATIONS FOR COMPOUND LIBRARY SCREENING 123 Key Learning Points
123 4.1 Measures of Assay Performance
125 4.2 Measuring Initial Velocity
133 4.3 Balanced Assay Conditions
142 4.4 Order of Reagent Addition
146 4.5 Use of Natural Substrates and Enzymes
148 4.6 Coupled Enzyme Assays
154 4.7 Hit Validation
156 4.8 Summary
166 References
166 5. LEAD OPTIMIZATION AND STRUCTURE-ACTIVITY RELATIONSHIPS FOR REVERSIBLE INHIBITORS 169 Key Learning Points
169 5.1 Concentration-Response Plots and IC50 Determination
170 5.2 Testing for Reversibility
183 5.3 Determining Reversible Inhibition Modality and Dissociation Constant
188 5.4 Comparing Relative Affinity
190 5.5 Associating Cellular Effects with Target Enzyme Inhibition
193 5.6 Summary
200 References
200 6. SLOW BINDING INHIBITORS 203 Key Learning Points
203 6.1 Determining kobs: The Rate Constant for Onset of Inhibition
205 6.2 Mechanisms of Slow Binding Inhibition
207 6.3 Determination of Mechanism and Assessment of True Affi nity
210 6.4 Determining Inhibition Modality for Slow Binding Inhibitors
217 6.5 SAR for Slow Binding Inhibitors
219 6.6 Some Examples of Pharmacologically Interesting Slow Binding Inhibitors
220 6.7 Summary
242 References
243 7. TIGHT BINDING INHIBITION 245 Key Learning Points
245 7.1 Effects of Tight Binding Inhibition on Concentration-Response Data
246 7.2 The IC50 Value Depends on Ki app and [E]T
248 7.3 Morrison's Quadratic Equation for Fitting Concentration-Response Data for Tight Binding Inhibitors
253 7.4 Determining Modality for Tight Binding Enzyme Inhibitors
258 7.5 Tight Binding Inhibitors Often Display Slow Binding Behavior
261 7.6 Practical Approaches to Overcoming the Tight Binding Limit in Determining Ki
263 7.7 Enzyme-Reaction Intermediate Analogues as Examples of Tight Binding Inhibitors
266 7.8 Potential Clinical Advantages of Tight Binding Inhibitors
277 7.9 Determination of [E]T Using Tight Binding Inhibitors
279 7.10 Summary
282 References
282 8. DRUG-TARGET RESIDENCE TIME 287 Key Learning Points
287 8.1 Open and Closed Systems in Biology
288 8.2 The Static View of Drug-Target Interactions
292 8.3 Conformational Adaptation in Drug-Target Interactions
294 8.4 Impact of Residence Time on Natural Receptor-Ligand Function
300 8.5 Impact of Drug-Target Residence Time on Drug Action
304 8.6 Experimental Measures of Drug-Target Residence Time
318 8.7 Drug-Target Residence Time Structure-Activity Relationships
325 8.8 Recent Applications of the Residence Time Concept
334 8.9 Limitations of Drug-Target Residence Time
338 8.10 Summary
340 References
341 9. IRREVERSIBLE ENZYME INACTIVATORS 345 Key Learning Points
345 9.1 Kinetic Evaluation of Irreversible Enzyme Inactivators
346 9.2 Affinity Labels
350 9.3 Mechanism-Based Inactivators
358 9.4 Use of Affi nity Labels as Mechanistic Tools
375 9.5 Summary
380 References
380 10. QUANTITATIVE BIOCHEMISTRY IN THE PHARMACOLOGICAL EVALUATION OF DRUGS 383 Key Learning Points
383 10.1 In Vitro ADMET Properties
384 10.2 In Vivo Pharmacokinetic Studies
426 10.3 Metabolite Identifi cation
453 10.4 Measures of Target Occupancy
454 10.5 Summary
465 References
466 APPENDIX 1 KINETICS OF BIOCHEMICAL REACTIONS 471 A1.1 The Law of Mass Action and Reaction Order
471 A1.2 First-Order Reaction Kinetics
475 A1.3 Second-Order Reaction Kinetics
478 A1.4 Pseudo-First-Order Reaction Conditions
479 A1.5 Approach to Equilibrium: An Example of the Kinetics of Reversible Reactions
480 APPENDIX 2 DERIVATION OF THE ENZYME-LIGAND BINDING ISOTHERM EQUATION 483 APPENDIX 3 SERIAL DILUTION SCHEMES 487 APPENDIX 4 RELATIONSHIP BETWEEN [I ]
IC50 AND PERCENTAGE INHIBITION OF ENZYME ACTIVITY WHEN h = 1 491 APPENDIX 5 PROPAGATION OF UNCERTAINTIES IN EXPERIMENTAL MEASUREMENTS 493 A5.1 Uncertainty Propagation for Addition or Subtraction of Two Experimental Parameters
493 A5.2 Uncertainty Propagation for Multiplication or Division of Two Experimental Parameters
494 A5.3 Uncertainty Propagation for Multiplication or Division of an Experimental Parameter by A Constant
494 A5.4 Uncertainty Propagation for an Experimental Parameter Raised by an Exponent
494 A5.5 Uncertainty Propagation for a General Function of Experimental Parameters
494 Reference
495 APPENDIX 6 USEFUL PHYSICAL CONSTANTS AT DIFFERENT TEMPERATURES 497 APPENDIX 7 COMMON RADIOACTIVE ISOTOPES USED IN STUDIES OF ENZYMES 499 APPENDIX 8 COMMON PREFIXES FOR UNITS IN BIOCHEMISTRY 501 APPENDIX 9 SOME AROMATIC RING SYSTEMS COMMONLY FOUND IN DRUGS 503 APPENDIX 10 RESIDUAL PLOTS 505 INDEX 509