Liang Shen, PhD, is the head of DMPK at WuXi AppTec.

• About the Editors ixPreface xiAcknowledgments xiiiSection I Novel Drug Modalities: Advancements and Their Pharmacokinetic Strategies 11 Proteolysis-Targeting Chimeras (PROTACs) 31.1 An Overview of PROTAC Technology and Drug Metabolism and Pharmacokinetic (DMPK) Research Strategies 31.2 Strategies to Improve the Oral Bioavailability of PROTACs 91.3 Research on PROTAC Metabolism: Strategies and Main Approaches 171.4 Metabolic Characteristics and Research Strategies for Metabolite Identification of PROTACs 231.5 Evaluation of Drug-Drug Interactions of PROTACs with Efflux Transporters 401.6 PROTAC Bioanalysis: Challenges and Strategies 442 Antibody–Drug Conjugates (ADCs) 492.1 General Considerations for Pharmacokinetic Studies of ADCs 492.2 Application of Integrated Bioanalysis in ADC DMPK Studies 532.3 Study Models and Bioanalysis Methods for the Payload-Related Metabolite Identification Studies of ADCs 622.4 Strategies and Challenges in the Determination of Drug-to-Antibody Ratio (DAR) Values for ADCs 672.5 In Vitro Stability of ADCs in Blood Matrices 782.6 Application of Radiolabeling Techniques in ADC Pharmacokinetic Studies 863 Peptide–Drug Conjugates (PDCs) 953.1 Introduction to PDCs and DMPK Research Strategies 953.2 Targeted Release Principle and Metabolite Identification Studies of PDCs 1024 Peptide Drugs 1154.1 Development, Characteristics, and Pharmacokinetic Research Strategies of Peptide Drugs 1154.2 Metabolic Characteristics and Optimization Strategies of Peptide Drugs 1234.3 Insights into ADME Studies of 37 Approved Therapeutic Peptides: Strategies and Radiolabeling Considerations 1324.4 Challenges and Strategies in the Bioanalysis of Peptide Drugs 1385 Oligonucleotide (OLIGO) Drugs 1475.1 Oligonucleotide Drugs: An Introduction and Overview of Pharmacokinetic Strategies 1475.2 Delivery Systems of siRNA Drugs and Their Effects on the Pharmacokinetic Profile 1525.3 Bioanalytical Strategies for Oligonucleotide Drugs 1575.4 Oligonucleotide Drugs: Strategies for Metabolism and Metabolite Profiling and Identification 1835.5 ADME Characteristics of Approved siRNA and ASO Drugs 1936 mRNA-Based Vaccines and Therapeutics 2136.1 What Are mRNA Vaccines and Their Future 2136.2 Strategies for Preclinical PK Evaluation of mRNA-Based Therapeutics: mRNA Vaccines, mRNA Therapeutic Drugs, and Novel Excipients 2216.3 FDA-Approved mRNA Vaccines: Interpretation of Preclinical Pharmacokinetic (PK) Data 2306.4 Application of Integrative Bioanalysis in mRNA Drugs DMPK Research 235Section II Pharmacokinetic and Metabolism Studies of Therapeutics for Disease Areas 2437 Ophthalmic Drugs 2457.1 Ophthalmic Drugs: Preclinical Pharmacokinetic (PK) Profiles and Research Strategies 2457.2 In Vitro Methods and Significance of Drug–Melanin Binding 2537.3 Preclinical Pharmacokinetic Research of Ophthalmic Drugs: In Vitro and In Vivo Research Methods 2597.4 In Vivo Bioanalysis in Ophthalmology: Challenges and Strategies 2698 Respiratory Drugs 2758.1 Inhaled Medications: Challenges and Strategies for Preclinical In Vivo Pharmacokinetic Studies 2758.2 Chronic Obstructive Pulmonary Disease (COPD): Analysis of PK Profiles of Approved Inhaled Medications 2859 Transdermal and Topical Drugs 2919.1 Preclinical Pharmacokinetic Strategies for Topical and Transdermal Drug Products 2919.2 Application of In Vitro Permeation Test (IVPT) for the Development of Transdermal and Topical Drugs 3009.3 Evaluation of Transdermal Drugs in In Vivo Pharmacokinetic Studies 3089.4 Challenges and Bioanalytical Strategies for Quantitative Detection of In Vivo Biological Samples from Skin Administration 31310 Central Nervous System Drugs 31710.1 Overview of Pharmacokinetic Research Strategies for Central Nervous System Drugs 31710.2 Application of Microdialysis in Pharmacokinetic Research 32910.3 Central Nervous System Drug Development Techniques: Exploration and Application in Large Animal Pharmacokinetics 337Section III Research Strategies, Methods, and Applications in Pharmacokinetics and Metabolism 34311 In Vitro ADME 34511.1 Rapid Determination of Lipophilicity: Establishment and Application of Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) 34511.2 Plasma Protein Binding Measurement of High Protein-Binding Drugs: Principlesand Advantages of the Flux Dialysis Method 35111.3 How to Measure the Metabolic Clearance of Slowly Metabolized Compounds 35811.4 Non-P450 Enzyme-Mediated Metabolism and In Vitro Assessment Method 36511.5 In Vitro Evaluation of CYP450 Time-Dependent Inhibition (TDI): A Novel Area Under the Curve Shift (AUC Shift) Approach 37711.6 Cytochrome P450 Reaction Phenotyping for Four Additional CYPs: Why and How 38311.7 Overview of the Application of the Na + -Taurocholate Cotransporting Polypeptide (NTCP) in Drug Development 38811.8 The Significance and In Vitro Methods of Lysosomal Trapping in Drug Development 39712 In Vivo Pharmacokinetics 40712.1 Enhancing the Bioavailability of Poorly Soluble Compounds: Strategies for Formulation Optimization in Preclinical Studies 40712.2 Self-Emulsifying Drug Delivery System (SEDDS): Enhancing the Oral Absorption of Lipophilic Compounds 41412.3 Application of Anesthetics in Rodent Pharmacokinetic Studies 42012.4 Application of In Situ Single-Pass Intestinal Perfusion (SPIP) Model in the Study of Intestinal Absorption of Oral Drugs 42812.5 Application of Liver Biopsy in Preclinical Pharmacokinetic Studies 43613 Metabolite Profiling and Identification 44113.1 High-Throughput Identification of Drug Metabolic Soft Spots for Lead Optimization 44113.2 Application of Metabolite Biosynthesis Technology in the Precise Identification of Drug Metabolite Structures 44913.3 Detection of Reactive Metabolites and Its Application in Drug Research and Development 45613.4 UGT Enzymes: Metabolite Identification and Toxicity Assessment Strategies 46613.5 Insights into the R&D Strategies of Deuterated Drugs from the Drug Metabolism Perspective 47214 Radiolabeled In Vivo ADME Studies 47914.1 Placental Transfer and Milk Excretion Studies Based on Radioactive Tracer Technique 47914.2 Visualization of Tissue Distribution: Quantitative Whole-Body Autoradiography (qwba) 48614.3 Preclinical and Clinical Research Strategy for Human Radiolabeled Mass Balance Studies 49014.4 Key Considerations for Conducting Radiolabeled Human Mass Balance Studies 49615 Bioanalysis 50315.1 Application of High-Throughput Mass Spectrometry in In Vitro ADME 50315.2 Application and Detection Techniques of Biomarkers in Drug Development 51015.3 Analysis and Detection Strategies of Liposome Drugs in Biological Samples 51615.4 DMPK Research Strategies of Chiral Drugs Based on UPCC-MS/MS and UPLC-MS/MS Dual Analytical Platforms 52315.5 Integrated Qualitative and Quantitative Bioanalysis of Intact Protein with High-Resolution Mass Spectrometry 53015.6 Nanobodies: Kinetics and Bioanalytical Strategies 540Index 549

Liang Shen, PhD, is the head of DMPK at WuXi AppTec.

• About the Editors ixPreface xiAcknowledgments xiiiSection I Novel Drug Modalities: Advancements and Their Pharmacokinetic Strategies 11 Proteolysis-Targeting Chimeras (PROTACs) 31.1 An Overview of PROTAC Technology and Drug Metabolism and Pharmacokinetic (DMPK) Research Strategies 31.2 Strategies to Improve the Oral Bioavailability of PROTACs 91.3 Research on PROTAC Metabolism: Strategies and Main Approaches 171.4 Metabolic Characteristics and Research Strategies for Metabolite Identification of PROTACs 231.5 Evaluation of Drug-Drug Interactions of PROTACs with Efflux Transporters 401.6 PROTAC Bioanalysis: Challenges and Strategies 442 Antibody–Drug Conjugates (ADCs) 492.1 General Considerations for Pharmacokinetic Studies of ADCs 492.2 Application of Integrated Bioanalysis in ADC DMPK Studies 532.3 Study Models and Bioanalysis Methods for the Payload-Related Metabolite Identification Studies of ADCs 622.4 Strategies and Challenges in the Determination of Drug-to-Antibody Ratio (DAR) Values for ADCs 672.5 In Vitro Stability of ADCs in Blood Matrices 782.6 Application of Radiolabeling Techniques in ADC Pharmacokinetic Studies 863 Peptide–Drug Conjugates (PDCs) 953.1 Introduction to PDCs and DMPK Research Strategies 953.2 Targeted Release Principle and Metabolite Identification Studies of PDCs 1024 Peptide Drugs 1154.1 Development, Characteristics, and Pharmacokinetic Research Strategies of Peptide Drugs 1154.2 Metabolic Characteristics and Optimization Strategies of Peptide Drugs 1234.3 Insights into ADME Studies of 37 Approved Therapeutic Peptides: Strategies and Radiolabeling Considerations 1324.4 Challenges and Strategies in the Bioanalysis of Peptide Drugs 1385 Oligonucleotide (OLIGO) Drugs 1475.1 Oligonucleotide Drugs: An Introduction and Overview of Pharmacokinetic Strategies 1475.2 Delivery Systems of siRNA Drugs and Their Effects on the Pharmacokinetic Profile 1525.3 Bioanalytical Strategies for Oligonucleotide Drugs 1575.4 Oligonucleotide Drugs: Strategies for Metabolism and Metabolite Profiling and Identification 1835.5 ADME Characteristics of Approved siRNA and ASO Drugs 1936 mRNA-Based Vaccines and Therapeutics 2136.1 What Are mRNA Vaccines and Their Future 2136.2 Strategies for Preclinical PK Evaluation of mRNA-Based Therapeutics: mRNA Vaccines, mRNA Therapeutic Drugs, and Novel Excipients 2216.3 FDA-Approved mRNA Vaccines: Interpretation of Preclinical Pharmacokinetic (PK) Data 2306.4 Application of Integrative Bioanalysis in mRNA Drugs DMPK Research 235Section II Pharmacokinetic and Metabolism Studies of Therapeutics for Disease Areas 2437 Ophthalmic Drugs 2457.1 Ophthalmic Drugs: Preclinical Pharmacokinetic (PK) Profiles and Research Strategies 2457.2 In Vitro Methods and Significance of Drug–Melanin Binding 2537.3 Preclinical Pharmacokinetic Research of Ophthalmic Drugs: In Vitro and In Vivo Research Methods 2597.4 In Vivo Bioanalysis in Ophthalmology: Challenges and Strategies 2698 Respiratory Drugs 2758.1 Inhaled Medications: Challenges and Strategies for Preclinical In Vivo Pharmacokinetic Studies 2758.2 Chronic Obstructive Pulmonary Disease (COPD): Analysis of PK Profiles of Approved Inhaled Medications 2859 Transdermal and Topical Drugs 2919.1 Preclinical Pharmacokinetic Strategies for Topical and Transdermal Drug Products 2919.2 Application of In Vitro Permeation Test (IVPT) for the Development of Transdermal and Topical Drugs 3009.3 Evaluation of Transdermal Drugs in In Vivo Pharmacokinetic Studies 3089.4 Challenges and Bioanalytical Strategies for Quantitative Detection of In Vivo Biological Samples from Skin Administration 31310 Central Nervous System Drugs 31710.1 Overview of Pharmacokinetic Research Strategies for Central Nervous System Drugs 31710.2 Application of Microdialysis in Pharmacokinetic Research 32910.3 Central Nervous System Drug Development Techniques: Exploration and Application in Large Animal Pharmacokinetics 337Section III Research Strategies, Methods, and Applications in Pharmacokinetics and Metabolism 34311 In Vitro ADME 34511.1 Rapid Determination of Lipophilicity: Establishment and Application of Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) 34511.2 Plasma Protein Binding Measurement of High Protein-Binding Drugs: Principlesand Advantages of the Flux Dialysis Method 35111.3 How to Measure the Metabolic Clearance of Slowly Metabolized Compounds 35811.4 Non-P450 Enzyme-Mediated Metabolism and In Vitro Assessment Method 36511.5 In Vitro Evaluation of CYP450 Time-Dependent Inhibition (TDI): A Novel Area Under the Curve Shift (AUC Shift) Approach 37711.6 Cytochrome P450 Reaction Phenotyping for Four Additional CYPs: Why and How 38311.7 Overview of the Application of the Na + -Taurocholate Cotransporting Polypeptide (NTCP) in Drug Development 38811.8 The Significance and In Vitro Methods of Lysosomal Trapping in Drug Development 39712 In Vivo Pharmacokinetics 40712.1 Enhancing the Bioavailability of Poorly Soluble Compounds: Strategies for Formulation Optimization in Preclinical Studies 40712.2 Self-Emulsifying Drug Delivery System (SEDDS): Enhancing the Oral Absorption of Lipophilic Compounds 41412.3 Application of Anesthetics in Rodent Pharmacokinetic Studies 42012.4 Application of In Situ Single-Pass Intestinal Perfusion (SPIP) Model in the Study of Intestinal Absorption of Oral Drugs 42812.5 Application of Liver Biopsy in Preclinical Pharmacokinetic Studies 43613 Metabolite Profiling and Identification 44113.1 High-Throughput Identification of Drug Metabolic Soft Spots for Lead Optimization 44113.2 Application of Metabolite Biosynthesis Technology in the Precise Identification of Drug Metabolite Structures 44913.3 Detection of Reactive Metabolites and Its Application in Drug Research and Development 45613.4 UGT Enzymes: Metabolite Identification and Toxicity Assessment Strategies 46613.5 Insights into the R&D Strategies of Deuterated Drugs from the Drug Metabolism Perspective 47214 Radiolabeled In Vivo ADME Studies 47914.1 Placental Transfer and Milk Excretion Studies Based on Radioactive Tracer Technique 47914.2 Visualization of Tissue Distribution: Quantitative Whole-Body Autoradiography (qwba) 48614.3 Preclinical and Clinical Research Strategy for Human Radiolabeled Mass Balance Studies 49014.4 Key Considerations for Conducting Radiolabeled Human Mass Balance Studies 49615 Bioanalysis 50315.1 Application of High-Throughput Mass Spectrometry in In Vitro ADME 50315.2 Application and Detection Techniques of Biomarkers in Drug Development 51015.3 Analysis and Detection Strategies of Liposome Drugs in Biological Samples 51615.4 DMPK Research Strategies of Chiral Drugs Based on UPCC-MS/MS and UPLC-MS/MS Dual Analytical Platforms 52315.5 Integrated Qualitative and Quantitative Bioanalysis of Intact Protein with High-Resolution Mass Spectrometry 53015.6 Nanobodies: Kinetics and Bioanalytical Strategies 540Index 549

Liang Shen, PhD, is the head of DMPK at WuXi AppTec.

• About the Editors ixPreface xiAcknowledgments xiiiSection I Novel Drug Modalities: Advancements and Their Pharmacokinetic Strategies 11 Proteolysis-Targeting Chimeras (PROTACs) 31.1 An Overview of PROTAC Technology and Drug Metabolism and Pharmacokinetic (DMPK) Research Strategies 31.2 Strategies to Improve the Oral Bioavailability of PROTACs 91.3 Research on PROTAC Metabolism: Strategies and Main Approaches 171.4 Metabolic Characteristics and Research Strategies for Metabolite Identification of PROTACs 231.5 Evaluation of Drug-Drug Interactions of PROTACs with Efflux Transporters 401.6 PROTAC Bioanalysis: Challenges and Strategies 442 Antibody–Drug Conjugates (ADCs) 492.1 General Considerations for Pharmacokinetic Studies of ADCs 492.2 Application of Integrated Bioanalysis in ADC DMPK Studies 532.3 Study Models and Bioanalysis Methods for the Payload-Related Metabolite Identification Studies of ADCs 622.4 Strategies and Challenges in the Determination of Drug-to-Antibody Ratio (DAR) Values for ADCs 672.5 In Vitro Stability of ADCs in Blood Matrices 782.6 Application of Radiolabeling Techniques in ADC Pharmacokinetic Studies 863 Peptide–Drug Conjugates (PDCs) 953.1 Introduction to PDCs and DMPK Research Strategies 953.2 Targeted Release Principle and Metabolite Identification Studies of PDCs 1024 Peptide Drugs 1154.1 Development, Characteristics, and Pharmacokinetic Research Strategies of Peptide Drugs 1154.2 Metabolic Characteristics and Optimization Strategies of Peptide Drugs 1234.3 Insights into ADME Studies of 37 Approved Therapeutic Peptides: Strategies and Radiolabeling Considerations 1324.4 Challenges and Strategies in the Bioanalysis of Peptide Drugs 1385 Oligonucleotide (OLIGO) Drugs 1475.1 Oligonucleotide Drugs: An Introduction and Overview of Pharmacokinetic Strategies 1475.2 Delivery Systems of siRNA Drugs and Their Effects on the Pharmacokinetic Profile 1525.3 Bioanalytical Strategies for Oligonucleotide Drugs 1575.4 Oligonucleotide Drugs: Strategies for Metabolism and Metabolite Profiling and Identification 1835.5 ADME Characteristics of Approved siRNA and ASO Drugs 1936 mRNA-Based Vaccines and Therapeutics 2136.1 What Are mRNA Vaccines and Their Future 2136.2 Strategies for Preclinical PK Evaluation of mRNA-Based Therapeutics: mRNA Vaccines, mRNA Therapeutic Drugs, and Novel Excipients 2216.3 FDA-Approved mRNA Vaccines: Interpretation of Preclinical Pharmacokinetic (PK) Data 2306.4 Application of Integrative Bioanalysis in mRNA Drugs DMPK Research 235Section II Pharmacokinetic and Metabolism Studies of Therapeutics for Disease Areas 2437 Ophthalmic Drugs 2457.1 Ophthalmic Drugs: Preclinical Pharmacokinetic (PK) Profiles and Research Strategies 2457.2 In Vitro Methods and Significance of Drug–Melanin Binding 2537.3 Preclinical Pharmacokinetic Research of Ophthalmic Drugs: In Vitro and In Vivo Research Methods 2597.4 In Vivo Bioanalysis in Ophthalmology: Challenges and Strategies 2698 Respiratory Drugs 2758.1 Inhaled Medications: Challenges and Strategies for Preclinical In Vivo Pharmacokinetic Studies 2758.2 Chronic Obstructive Pulmonary Disease (COPD): Analysis of PK Profiles of Approved Inhaled Medications 2859 Transdermal and Topical Drugs 2919.1 Preclinical Pharmacokinetic Strategies for Topical and Transdermal Drug Products 2919.2 Application of In Vitro Permeation Test (IVPT) for the Development of Transdermal and Topical Drugs 3009.3 Evaluation of Transdermal Drugs in In Vivo Pharmacokinetic Studies 3089.4 Challenges and Bioanalytical Strategies for Quantitative Detection of In Vivo Biological Samples from Skin Administration 31310 Central Nervous System Drugs 31710.1 Overview of Pharmacokinetic Research Strategies for Central Nervous System Drugs 31710.2 Application of Microdialysis in Pharmacokinetic Research 32910.3 Central Nervous System Drug Development Techniques: Exploration and Application in Large Animal Pharmacokinetics 337Section III Research Strategies, Methods, and Applications in Pharmacokinetics and Metabolism 34311 In Vitro ADME 34511.1 Rapid Determination of Lipophilicity: Establishment and Application of Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) 34511.2 Plasma Protein Binding Measurement of High Protein-Binding Drugs: Principlesand Advantages of the Flux Dialysis Method 35111.3 How to Measure the Metabolic Clearance of Slowly Metabolized Compounds 35811.4 Non-P450 Enzyme-Mediated Metabolism and In Vitro Assessment Method 36511.5 In Vitro Evaluation of CYP450 Time-Dependent Inhibition (TDI): A Novel Area Under the Curve Shift (AUC Shift) Approach 37711.6 Cytochrome P450 Reaction Phenotyping for Four Additional CYPs: Why and How 38311.7 Overview of the Application of the Na + -Taurocholate Cotransporting Polypeptide (NTCP) in Drug Development 38811.8 The Significance and In Vitro Methods of Lysosomal Trapping in Drug Development 39712 In Vivo Pharmacokinetics 40712.1 Enhancing the Bioavailability of Poorly Soluble Compounds: Strategies for Formulation Optimization in Preclinical Studies 40712.2 Self-Emulsifying Drug Delivery System (SEDDS): Enhancing the Oral Absorption of Lipophilic Compounds 41412.3 Application of Anesthetics in Rodent Pharmacokinetic Studies 42012.4 Application of In Situ Single-Pass Intestinal Perfusion (SPIP) Model in the Study of Intestinal Absorption of Oral Drugs 42812.5 Application of Liver Biopsy in Preclinical Pharmacokinetic Studies 43613 Metabolite Profiling and Identification 44113.1 High-Throughput Identification of Drug Metabolic Soft Spots for Lead Optimization 44113.2 Application of Metabolite Biosynthesis Technology in the Precise Identification of Drug Metabolite Structures 44913.3 Detection of Reactive Metabolites and Its Application in Drug Research and Development 45613.4 UGT Enzymes: Metabolite Identification and Toxicity Assessment Strategies 46613.5 Insights into the R&D Strategies of Deuterated Drugs from the Drug Metabolism Perspective 47214 Radiolabeled In Vivo ADME Studies 47914.1 Placental Transfer and Milk Excretion Studies Based on Radioactive Tracer Technique 47914.2 Visualization of Tissue Distribution: Quantitative Whole-Body Autoradiography (qwba) 48614.3 Preclinical and Clinical Research Strategy for Human Radiolabeled Mass Balance Studies 49014.4 Key Considerations for Conducting Radiolabeled Human Mass Balance Studies 49615 Bioanalysis 50315.1 Application of High-Throughput Mass Spectrometry in In Vitro ADME 50315.2 Application and Detection Techniques of Biomarkers in Drug Development 51015.3 Analysis and Detection Strategies of Liposome Drugs in Biological Samples 51615.4 DMPK Research Strategies of Chiral Drugs Based on UPCC-MS/MS and UPLC-MS/MS Dual Analytical Platforms 52315.5 Integrated Qualitative and Quantitative Bioanalysis of Intact Protein with High-Resolution Mass Spectrometry 53015.6 Nanobodies: Kinetics and Bioanalytical Strategies 540Index 549