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| Foreword | |
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| Preface | |
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| Acknowledgments | |
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| Why Enzymes as Drug Targets? | |
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| Enzymes Are Essentials for Life | |
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| Enzyme Structure and Catalysis | |
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| Permutations of Enzyme Structure During Catalysis | |
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| Other Reasons for Studying Enzymes | |
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| Summary | |
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| References | |
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| Enzyme Reaction Mechanisms | |
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| Initial Binding of Substrate | |
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| Noncovalent Forces in Reversible Ligand Binding to Enzymes | |
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| Electrostatic Forces | |
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| Hydrogen Bonds | |
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| Hydrophobic Forces | |
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| van der Waals Forces | |
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| Transformations of the Bond Substrate | |
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| Strategies for Transition State Stabilization | |
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| Enzyme Active Sites Are Most Complementary to the Transition State Structure | |
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| Steady State Analysis of Enzyme Kinetics | |
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| Factors Affecting the Steady State Kinetic Constants | |
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| Graphical Determination ofk cat and K M | |
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| Reactions Involving Multiple Substates | |
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| Bisubstrate Reaction Mechanisms | |
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| Summary | |
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| References | |
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| Reversible Modes of Inhibitor Interactions with Enzymes | |
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| Enzyme-Inhibitor Binding Equilibria | |
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| Competitive Inhibition | |
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| Noncompetitive Inhibition | |
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| Mutual Exclusively Studies | |
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| Uncompetitive Inhibition | |
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| Inhibition Modality in Bisubstrate Reactions | |
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| Value of Knowing Inhibitor Modality | |
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| Quantitative Comparisons of Inhibitor Affinity | |
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| RelatingK i to Binding Energy | |
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| Defining Target Selectivity byK i Values | |
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| Potential Advantages and Disadvantages of Different Inhibition Modalities In Vivo | |
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| Knowing Inhibition Modality Is Important for Structure-Based Lead Organization | |
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| Summary | |
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| References | |
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| Assay Considerations for Compound Library Screening | |
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| Defining Inhibition Signal Robustness, and Hit Criteria | |
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| Measuring Initial Velocity | |
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| End-Point and Kinetic Readouts | |
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| Effects of Enzyme Concentration | |
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| Balanced Assay Conditions | |
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| Balancing Conditions for Multisubstrate Reactions | |
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| Order of Reagent Addition | |
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| Use of Natural Substrates and Enzymes | |
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| Coupled Enzyme Assays | |
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| Hit Validation and Progression | |
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| Summary | |
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| References | |
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| Lead Optimization and Structure-Activity Relationships for Reversible Inhibitors | |
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| Concentration-Response Plots and IC 50 Determination | |
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| The Hill Coefficient | |
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| Graphing and Reporting Concentration-Response Data | |
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| Testing for Reversibility | |
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| Determining Reversible Inhibition Modality and Dissociation Constant | |
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| Comparing Relative Affinity | |
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| Compound Selectivity | |
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| Associating Cellular Effects with Target Enzyme Inhibition | |
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| Cellular Phenotype Should Be Consistent with Genetic Knockout or Knockdown of the Target Enzyme | |
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| Cellular Activity Should Require a Certain Affinity for the target Enzyme | |
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| Buildup of Substrate and/or Diminution of Product for the Target Enzyme Should Be Observed in Cells | |
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| Cellular Phenotype Should Be Reversed by Cell-Permeable Product or Downstream Metabolites of the Target Enzyme Activity | |
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| Mutation of the Target Enzyme Should Lead to Resistance or Hypersensitivity to Inhibitors | |
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| Summary | |
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| References | |
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| Slow Binding Inhibitors | |
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| Determiningk obs : The Rate Constant for Onset of Inhibition | |
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| Mechanisms of Slow Binding Inhibition | |
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| Determination of Mechanism and Assessment of True Affinity | |
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| Potential Clinical Advantages of Slow Off-rate Inhibitors | |
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| Determining Inhibition Modality for Slow Binding Inhibitors | |
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| SAR for Slow Binding Inhibitors | |
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| Some Examples of Pharmacologically Interes | |