{"product_id":"oligonucleotide-based-drugs-and-therapeutics-isbn-9781118537336","title":"Oligonucleotide-Based Drugs and Therapeutics","description":"\u003cp\u003e\u003cb\u003eA comprehensive review of contemporary antisense oligonucleotides drugs and therapeutic principles, methods, applications, and research\u003c\/b\u003e \u003c\/p\u003e \u003cp\u003eOligonucleotide-based drugs, in particular antisense oligonucleotides, are part of a growing number of pharmaceutical and biotech programs progressing to treat a wide range of indications including cancer, cardiovascular, neurodegenerative, neuromuscular, and respiratory diseases, as well as other severe and rare diseases. Reviewing fundamentals and offering guidelines for drug discovery and development, this book is a practical guide covering all key aspects of this increasingly popular area of pharmacology and biotech and pharma research, from the basic science behind antisense oligonucleotides chemistry, toxicology, manufacturing, to safety assessments, the design of therapeutic protocols, to clinical experience.\u003c\/p\u003e \u003cp\u003eAntisense oligonucleotides are single strands of DNA or RNA that are complementary to a chosen sequence. While the idea of antisense oligonucleotides to target single genes dates back to the 1970's, most advances have taken place in recent years. The increasing number of antisense oligonucleotide programs in clinical development is a testament to the progress and understanding of pharmacologic, pharmacokinetic, and toxicologic properties as well as improvement in the delivery of oligonucleotides. This valuable book reviews the fundamentals of oligonucleotides, with a focus on antisense oligonucleotide drugs, and reports on the latest research underway worldwide.\u003c\/p\u003e \u003cp\u003e•    Helps readers understand antisense molecules and their targets, biochemistry, and toxicity mechanisms, roles in disease, and applications for safety and therapeutics\u003c\/p\u003e \u003cp\u003e•    Examines the principles, practices, and tools for scientists in both pre-clinical and clinical settings and how to apply them to antisense oligonucleotides\u003c\/p\u003e \u003cp\u003e•    Provides guidelines for scientists in drug design and discovery to help improve efficiency, assessment, and the success of drug candidates\u003c\/p\u003e \u003cp\u003e•    Includes interdisciplinary perspectives, from academia, industry, regulatory and from the fields of pharmacology, toxicology, biology, and medicinal chemistry\u003c\/p\u003e \u003cp\u003e\u003ci\u003eOligonucleotide-Based Drugs and Therapeutics\u003c\/i\u003e belongs on the reference shelves of chemists, pharmaceutical scientists, chemical biologists, toxicologists and other scientists working in the pharmaceutical and biotechnology industries. It will also be a valuable resource for regulatory specialists and safety assessment professionals and an important reference for academic researchers and post-graduates interested in therapeutics, antisense therapy, and oligonucleotides.\u003c\/p\u003e \u003cp\u003eList of Contributors xvii\u003c\/p\u003e \u003cp\u003ePreface xxi\u003c\/p\u003e \u003cp\u003eAcknowledgments xxii\u003c\/p\u003e \u003cp\u003e\u003cb\u003e1 Mechanisms of Oligonucleotide Actions 1\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eAnnemieke Aartsma‐Rus, Aimee L. Jackson, and Arthur A. Levin\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e1.1 Introduction\u003c\/p\u003e \u003cp\u003e1.2 Antisense Oligonucleotide Therapeutics 2\u003c\/p\u003e \u003cp\u003e1.2.1 Antisense Activity Mediated by RNase H 2\u003c\/p\u003e \u003cp\u003e1.2.2 The RNase H Mechanism 2\u003c\/p\u003e \u003cp\u003e1.2.3 Chemical Modifications to Enhance RNase H‐mediated Antisense Activity 3\u003c\/p\u003e \u003cp\u003e1.3 Oligonucleotides that Sterically Block Translation 5\u003c\/p\u003e \u003cp\u003e1.4 Oligonucleotides that Act Through the RNAi Pathway 5\u003c\/p\u003e \u003cp\u003e1.4.1 The RISC Pathway 5\u003c\/p\u003e \u003cp\u003e1.4.2 Mechanisms of RISC‐mediated Gene Silencing 8\u003c\/p\u003e \u003cp\u003e1.5 Chemical Modification of siRNAs and miRNAs 10\u003c\/p\u003e \u003cp\u003e1.5.1 Delivery of Therapeutic siRNAs or miRNAs 12\u003c\/p\u003e \u003cp\u003e1.6 Clinical Use of Oligonucleotides that Act through the RNAi Pathway 14\u003c\/p\u003e \u003cp\u003e1.7 Oligonucleotides that Modulate Splicing 17\u003c\/p\u003e \u003cp\u003e1.7.1 Pre‐mRNA Splicing and Disease 17\u003c\/p\u003e \u003cp\u003e1.7.2 Mechanisms of Oligonucleotide‐mediated Splicing Modulation 17\u003c\/p\u003e \u003cp\u003e1.7.3 Chemical Modifications that Enhance Activity of Oligonucleotidebased Splicing Modulators 21\u003c\/p\u003e \u003cp\u003e1.7.4 Clinical Applications of Splicing Modulators 22\u003c\/p\u003e \u003cp\u003e1.8 Conclusions 22\u003c\/p\u003e \u003cp\u003eReferences 22\u003c\/p\u003e \u003cp\u003e\u003cb\u003e2 The Medicinal Chemistry of Antisense Oligonucleotides 39\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eJonathan K. Watts\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e2.1 Introduction:The Antisense Approach and the Need for Chemical Modification 39\u003c\/p\u003e \u003cp\u003e2.1.1 How Does Medicinal Chemistry Apply to Oligonucleotides? 40\u003c\/p\u003e \u003cp\u003e2.1.2 Chemistry and Toxicity 41\u003c\/p\u003e \u003cp\u003e2.2 Why Chemically Modify an Oligonucleotide? 42\u003c\/p\u003e \u003cp\u003e2.2.1 Medicinal Chemistry Can Increase Nuclease Stability 42\u003c\/p\u003e \u003cp\u003e2.2.2 Medicinal Chemistry Can Tune Binding Affinity and Specificity 43\u003c\/p\u003e \u003cp\u003e2.2.3 Medicinal Chemistry Can Change Interactions with Cellular Factors 44\u003c\/p\u003e \u003cp\u003e2.2.4 Medicinal Chemistry Can Modulate Immunostimulation 45\u003c\/p\u003e \u003cp\u003e2.2.5 Medicinal Chemistry Can Improve RNase H Cleavage Specificity 46\u003c\/p\u003e \u003cp\u003e2.2.6 Medicinal Chemistry Can Improve Cellular Uptake and Subcellular Trafficking 47\u003c\/p\u003e \u003cp\u003e2.3 Chemical Modifications of Current Importance by Structural Class 48\u003c\/p\u003e \u003cp\u003e2.3.1 Sugar Modifications 48\u003c\/p\u003e \u003cp\u003e2.3.1.1 2′‐Modified Ribose Sugars 48\u003c\/p\u003e \u003cp\u003e2.3.1.2 2′‐Modified Arabinose Sugars 50\u003c\/p\u003e \u003cp\u003e2.3.1.3 2′,4′‐Difluorinated Nucleosides 50\u003c\/p\u003e \u003cp\u003e2.3.1.4 Constrained Nucleotides 50\u003c\/p\u003e \u003cp\u003e2.3.1.5 Sugars with Expanded Ring Size 53\u003c\/p\u003e \u003cp\u003e2.3.2 Phosphate Modifications 54\u003c\/p\u003e \u003cp\u003e2.3.2.1 Phosphorothioate 54\u003c\/p\u003e \u003cp\u003e2.3.2.2 Other Charged Phosphate Analogues 58\u003c\/p\u003e \u003cp\u003e2.3.2.3 Neutral Mimics of the Phosphate Linkage 58\u003c\/p\u003e \u003cp\u003e2.3.2.4 Metabolically Stable 5′‐Phosphate Analogues 60\u003c\/p\u003e \u003cp\u003e2.3.3 Total Replacement of the Sugar‐Phosphate Backbone 61\u003c\/p\u003e \u003cp\u003e2.3.4 Nucleobase Modifications 62\u003c\/p\u003e \u003cp\u003e2.3.4.1 Sulfur‐Modified Nucleobases 63\u003c\/p\u003e \u003cp\u003e2.3.4.2 5‐Modified Pyrimidines 63\u003c\/p\u003e \u003cp\u003e2.3.4.3 Nucleobases with Expanded Hydrogen Bonding Networks 65\u003c\/p\u003e \u003cp\u003e2.3.5 Assembly of Oligonucleotides into Multimeric Structures 66\u003c\/p\u003e \u003cp\u003e2.4 Conclusion 67\u003c\/p\u003e \u003cp\u003eReferences 69\u003c\/p\u003e \u003cp\u003e\u003cb\u003e3 Cellular Pharmacology of Antisense Oligonucleotides 91\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eXin Ming\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e3.1 Introduction91\u003c\/p\u003e \u003cp\u003e3.2 Molecular Mechanisms of Antisense Oligonucleotides 92\u003c\/p\u003e \u003cp\u003e3.2.1 Classic Antisense Oligonucleotides 92\u003c\/p\u003e \u003cp\u003e3.2.2 siRNA 94\u003c\/p\u003e \u003cp\u003e3.2.3 Splice Switching Oligonucleotides 94\u003c\/p\u003e \u003cp\u003e3.2.4 microRNA Antagomirs 95\u003c\/p\u003e \u003cp\u003e3.2.5 lncRNAs Antagomirs 95\u003c\/p\u003e \u003cp\u003e3.3 Cellular Pharmacology of Antisense Oligonucleotides 96\u003c\/p\u003e \u003cp\u003e3.3.1 Endocytosis of Free Oligonucleotides 98\u003c\/p\u003e \u003cp\u003e3.3.2 Endocytosis of Oligonucleotide Conjugates 98\u003c\/p\u003e \u003cp\u003e3.3.3 Uptake and Trafficking of Oligonucleotides Incorporated into Nanocarriers 100\u003c\/p\u003e \u003cp\u003e3.4 Conclusion 101\u003c\/p\u003e \u003cp\u003eReferences 101\u003c\/p\u003e \u003cp\u003e\u003cb\u003e4 Pharmacokinetics and Pharmacodynamics of Antisense Oligonucleotides 107\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eHelen Lightfoot, Anneliese Schneider, and Jonathan Hall\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e4.1 Introduction 107\u003c\/p\u003e \u003cp\u003e4.2 Pharmacokinetic Properties of Antisense Oligonucleotides 108\u003c\/p\u003e \u003cp\u003e4.2.1 Protein Binding 109\u003c\/p\u003e \u003cp\u003e4.2.2 Dose Dependency of ASO Pharmacokinetics 110\u003c\/p\u003e \u003cp\u003e4.2.3 Absorption 110\u003c\/p\u003e \u003cp\u003e4.2.4 Distribution 111\u003c\/p\u003e \u003cp\u003e4.2.5 Metabolism and Excretion 112\u003c\/p\u003e \u003cp\u003e4.3 Pharmacodynamic Properties of Antisense Oligonucleotides 113\u003c\/p\u003e \u003cp\u003e4.3.1 ASO Target Selection and Validation 114\u003c\/p\u003e \u003cp\u003e4.3.2 Mechanisms of Action 117\u003c\/p\u003e \u003cp\u003e4.3.3 Biomarkers and PD Endpoints 118\u003c\/p\u003e \u003cp\u003e4.4 PD and PK Results and Strategies of ASOs in Clinical Development 119\u003c\/p\u003e \u003cp\u003e4.4.1 Genetic Diseases 122\u003c\/p\u003e \u003cp\u003e4.4.1.1 Mipomersen, Apolipoprotein B‐100, and Hypercholesterolemia 122\u003c\/p\u003e \u003cp\u003e4.4.1.2 Drisapersen, Dystrophin, and Duchenne Muscular Dystrophy (DMD) 123\u003c\/p\u003e \u003cp\u003e4.4.2 Infectious Diseases 125\u003c\/p\u003e \u003cp\u003e4.4.2.1 Miravirsen, miR‐122, and Hepatitis C Virus (HCV) 125\u003c\/p\u003e \u003cp\u003e4.4.3 Cancer 126\u003c\/p\u003e \u003cp\u003e4.4.3.1 Custirsen, Clusterin, and Cancer 126\u003c\/p\u003e \u003cp\u003e4.4.3.2 LY2181308 (ISIS‐23722), Survivin, and Cancer 127\u003c\/p\u003e \u003cp\u003e4.5 Summary and Conclusions 128\u003c\/p\u003e \u003cp\u003eReferences 130\u003c\/p\u003e \u003cp\u003e\u003cb\u003e5 Tissue Distribution, Metabolism, and Clearance 137\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eMehrdad Dirin and Johannes Winkler\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e5.1 Introduction137\u003c\/p\u003e \u003cp\u003e5.2 Tissue Distribution 138\u003c\/p\u003e \u003cp\u003e5.2.1 Dermal Delivery 138\u003c\/p\u003e \u003cp\u003e5.2.2 Ocular Delivery 139\u003c\/p\u003e \u003cp\u003e5.2.3 Oral Administration 139\u003c\/p\u003e \u003cp\u003e5.2.4 Intrathecal Delivery 141\u003c\/p\u003e \u003cp\u003e5.2.5 Intravesical Administration 142\u003c\/p\u003e \u003cp\u003e5.2.6 Pulmonary Administration 142\u003c\/p\u003e \u003cp\u003e5.2.7 Distribution to Muscular Tissue 143\u003c\/p\u003e \u003cp\u003e5.2.8 Intravenous Administration 144\u003c\/p\u003e \u003cp\u003e5.3 Cellular Uptake 146\u003c\/p\u003e \u003cp\u003e5.4 Metabolism and Clearance 148\u003c\/p\u003e \u003cp\u003e5.4.1 Phosphorothioates Including 2′‐Modifications 148\u003c\/p\u003e \u003cp\u003e5.4.2 Phosphorodiamidate Morpholino Oligonucleotides 149\u003c\/p\u003e \u003cp\u003e5.5 Conclusion 150\u003c\/p\u003e \u003cp\u003eReferences 151\u003c\/p\u003e \u003cp\u003e\u003cb\u003e6 Hybridization‐Independent Effects: Principles and Specific Considerations for Oligonucleotide Drugs 161\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eNicolay Ferrari\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e6.1 Background 161\u003c\/p\u003e \u003cp\u003e6.2 Mechanisms of Hybridization‐independent Toxicities 162\u003c\/p\u003e \u003cp\u003e6.2.1 Effects Related to Oligonucleotide Sequence 162\u003c\/p\u003e \u003cp\u003e6.2.1.1 Unmethylated CpG Motifs 162\u003c\/p\u003e \u003cp\u003e6.2.1.2 Poly‐G Sequences 163\u003c\/p\u003e \u003cp\u003e6.2.1.3 DNA Triplex‐forming Oligonucleotides 164\u003c\/p\u003e \u003cp\u003e6.2.1.4 Other Motifs 164\u003c\/p\u003e \u003cp\u003e6.2.2 Effects Related to Oligonucleotide Chemistry 164\u003c\/p\u003e \u003cp\u003e6.2.2.1 Phosphorothioate Oligonucleotides 165\u003c\/p\u003e \u003cp\u003e6.2.2.2 Effects of Other Chemical Modifications 171\u003c\/p\u003e \u003cp\u003e6.3 Hybridization‐independent Effects Following Local Delivery of Oligonucleotides 171\u003c\/p\u003e \u003cp\u003e6.3.1 Pulmonary Toxicity of Inhaled Oligonucleotides 171\u003c\/p\u003e \u003cp\u003e6.3.1.1 Specific Considerations for Inhaled Oligonucleotides 173\u003c\/p\u003e \u003cp\u003e6.3.2 Approaches to Reduce Hybridization‐independent Class Effects of Inhaled Oligonucleotides 175\u003c\/p\u003e \u003cp\u003e6.3.2.1 Mixed Phosphorothioate\/Phosphodiester Oligonucleotides 175\u003c\/p\u003e \u003cp\u003e6.4 Conclusion 180\u003c\/p\u003e \u003cp\u003eReferences 180\u003c\/p\u003e \u003cp\u003e\u003cb\u003e7 Hybridization‐Dependent Effects: The Prediction, Evaluation,and Consequences of Unintended Target Hybridization 191\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eJeremy D. A. Kitson, Piotr J. Kamola, and Lauren Kane\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e7.1 Introduction 191\u003c\/p\u003e \u003cp\u003e7.1.1 Scope of this Review: RNase H1‐dependent ASOs 192\u003c\/p\u003e \u003cp\u003e7.2 Specificity Studies with ASOs 192\u003c\/p\u003e \u003cp\u003e7.3 Implications of the Nuclear Site of Action of RNase H1 194\u003c\/p\u003e \u003cp\u003e7.3.1 Confirmation of Unintended Targets within Introns 195\u003c\/p\u003e \u003cp\u003e7.4 Mechanism of OTE 196\u003c\/p\u003e \u003cp\u003e7.5 Determining the Extent that Accessibility, Affinity and, Mismatch Tolerance Contribute to Off‐target Activity 198\u003c\/p\u003e \u003cp\u003e7.5.1 Accessibility 198\u003c\/p\u003e \u003cp\u003e7.5.2 Affinity 199\u003c\/p\u003e \u003cp\u003e7.5.3 The Interaction of RNase H1 with the RNA\/ASO Duplex 200\u003c\/p\u003e \u003cp\u003e7.5.4 Mismatch Tolerance 202\u003c\/p\u003e \u003cp\u003e7.6 Consequences of Unintended Transcript Knockdown: \u003ci\u003eIn Vivo\u003c\/i\u003e and \u003ci\u003eIn Vitro\u003c\/i\u003e Toxicity 203\u003c\/p\u003e \u003cp\u003e7.7 Identification and Evaluation of Putative OTEs 207\u003c\/p\u003e \u003cp\u003e7.7.1 Computational Prediction of Unintended Targeting 207\u003c\/p\u003e \u003cp\u003e7.7.1.1 Database Creation 209\u003c\/p\u003e \u003cp\u003e7.7.1.2 Sequence Alignments 209\u003c\/p\u003e \u003cp\u003e7.7.1.3 Cross‐species Off‐target Homology 210\u003c\/p\u003e \u003cp\u003e7.7.1.4 Results Filtering and Annotation 211\u003c\/p\u003e \u003cp\u003e7.7.1.5 RNA Structure and Target Accessibility 211\u003c\/p\u003e \u003cp\u003e7.7.1.6 ASO–Target Duplex Thermodynamics 213\u003c\/p\u003e \u003cp\u003e7.7.1.7 Computational Framework for OTEs 214\u003c\/p\u003e \u003cp\u003e7.7.1.8 \u003ci\u003eIn Vitro\u003c\/i\u003e Screening for OTEs 214\u003c\/p\u003e \u003cp\u003e7.7.1.9 Methods for Measuring Gene Expression 216\u003c\/p\u003e \u003cp\u003e7.8 Summary 216\u003c\/p\u003e \u003cp\u003eAcknowledgments 217\u003c\/p\u003e \u003cp\u003eReferences 218\u003c\/p\u003e \u003cp\u003e\u003cb\u003e8 Class‐Related Proinflammatory Effects 227\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eRosanne Seguin\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e8.1 Introduction 227\u003c\/p\u003e \u003cp\u003e8.2 Proinflammatory Effects of ASO for Consideration in Drug Development 228\u003c\/p\u003e \u003cp\u003e8.2.1 Activation of the Complement Cascade in Monkeys 228\u003c\/p\u003e \u003cp\u003e8.2.2 Cytokine Release 229\u003c\/p\u003e \u003cp\u003e8.2.3 Mononuclear Cellular Infiltrate 232\u003c\/p\u003e \u003cp\u003e8.2.4 Hematological Changes 236\u003c\/p\u003e \u003cp\u003e8.2.5 Immunogenicity 237\u003c\/p\u003e \u003cp\u003e8.3 Conclusions 238\u003c\/p\u003e \u003cp\u003eReferences 239\u003c\/p\u003e \u003cp\u003e\u003cb\u003e9 Exaggerated Pharmacology 243\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eAlain Guimond and Doug Kornbrust\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e9.1 Introduction 243\u003c\/p\u003e \u003cp\u003e9.2 Regulatory Expectations 244\u003c\/p\u003e \u003cp\u003e9.3 Scope of EP Assessment 245\u003c\/p\u003e \u003cp\u003e9.3.1 Species Selection 245\u003c\/p\u003e \u003cp\u003e9.3.2 Determination of Pharmacologic Relevance 247\u003c\/p\u003e \u003cp\u003e9.4 EP Evaluation Strategies 248\u003c\/p\u003e \u003cp\u003e9.4.1 Concerns About the Use of Animal‐active Analogues 248\u003c\/p\u003e \u003cp\u003e9.4.2 Animal‐active Analogues in Reproductive and\/or Carcinogenicity Studies 250\u003c\/p\u003e \u003cp\u003e9.4.3 Other Considerations for Use of Animal Analogues 250\u003c\/p\u003e \u003cp\u003e9.4.4 The Use of Inactive Analogues as Control Articles 250\u003c\/p\u003e \u003cp\u003e9.4.5 The Role of Formulations 251\u003c\/p\u003e \u003cp\u003e9.4.6 Aptamer Oligonucleotides 251\u003c\/p\u003e \u003cp\u003e9.4.7 Immunostimulatory Oligonucleotides 252\u003c\/p\u003e \u003cp\u003e9.4.8 MicroRNA 253\u003c\/p\u003e \u003cp\u003e9.5 Conclusions 254\u003c\/p\u003e \u003cp\u003eReferences 255\u003c\/p\u003e \u003cp\u003e\u003cb\u003e10 Genotoxicity Tests for Novel Oligonucleotide‐Based Therapeutics 257\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eCindy L. Berman, Scott A. Barros, Sheila M. Galloway, Peter Kasper, Frederick B. Oleson, Catherine C. Priestley, Kevin S. Sweder, Michael J. Schlosser, and Zhanna Sobol\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e10.1 Introduction 257\u003c\/p\u003e \u003cp\u003e10.1.1 History of Regulatory Guidance on Genotoxicity Testing 259\u003c\/p\u003e \u003cp\u003e10.1.2 Relevance of the Standard Genotoxicity Test Battery to ONs 260\u003c\/p\u003e \u003cp\u003e10.2 Experience with ONs in the Standard Battery 262\u003c\/p\u003e \u003cp\u003e10.2.1 ON Chemical Classes Tested for Genotoxicity 264\u003c\/p\u003e \u003cp\u003e10.2.2 Conclusions Based on the Database 265\u003c\/p\u003e \u003cp\u003e10.3 OSWG Recommendation for Genotoxicity Testing of ONs 266\u003c\/p\u003e \u003cp\u003e10.3.1 Recommended Test Battery 266\u003c\/p\u003e \u003cp\u003e10.3.2 Requirement for Evidence for Uptake 270\u003c\/p\u003e \u003cp\u003e10.3.3 Need for Testing of ONs 271\u003c\/p\u003e \u003cp\u003e10.3.3.1 Nonconjugated ONs in Simple Aqueous Formulations 271\u003c\/p\u003e \u003cp\u003e10.3.3.2 ONs in Complex Formulations or Conjugates 272\u003c\/p\u003e \u003cp\u003e10.3.4 Recommended Test Conditions 273\u003c\/p\u003e \u003cp\u003e10.3.4.1 Top Concentration for \u003ci\u003eIn Vitro\u003c\/i\u003e Tests 273\u003c\/p\u003e \u003cp\u003e10.3.4.2 Use of S‐9 in \u003ci\u003eIn Vitro\u003c\/i\u003e Tests 273\u003c\/p\u003e \u003cp\u003e10.3.4.3 In Vivo Tests 274\u003c\/p\u003e \u003cp\u003e10.4 Triplex Formation 275\u003c\/p\u003e \u003cp\u003e10.4.1 Biochemical Requirements for Triplex Formation 275\u003c\/p\u003e \u003cp\u003e10.4.2 Assessment of New ONs for Triplex Formation 277\u003c\/p\u003e \u003cp\u003e10.5 Impurities 278\u003c\/p\u003e \u003cp\u003e10.5.1 ON‐Related Impurities 278\u003c\/p\u003e \u003cp\u003e10.5.2 Potentially Mutagenic Impurities 278\u003c\/p\u003e \u003cp\u003e10.6 Conclusions 279\u003c\/p\u003e \u003cp\u003eAcknowledgments 280\u003c\/p\u003e \u003cp\u003eReferences 280\u003c\/p\u003e \u003cp\u003e\u003cb\u003e11 Reproductive and Developmental Toxicity Testing Strategies for Oligonucleotide‐Based Therapeutics 287\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eTacey E.K. White and Joy Cavagnaro\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e11.1 Introduction 287\u003c\/p\u003e \u003cp\u003e11.2 General Design of Reproductive and Developmental Toxicity Studies 289\u003c\/p\u003e \u003cp\u003e11.3 Product Attributes of Oligonucleotide Drugs 291\u003c\/p\u003e \u003cp\u003e11.4 The Role of Intended Pharmacology in Reproductive and Developmental Effects 293\u003c\/p\u003e \u003cp\u003e11.5 Selection of Animal Species 294\u003c\/p\u003e \u003cp\u003e11.5.1 Design and Use of Animal‐active Analogues 294\u003c\/p\u003e \u003cp\u003e11.6 Justification of Dosing Regimen 296\u003c\/p\u003e \u003cp\u003e11.7 Exposure Assessment 297\u003c\/p\u003e \u003cp\u003e11.8 Subclass‐ specific Considerations 298\u003c\/p\u003e \u003cp\u003e11.8.1 Single‐stranded DNA Antisense Oligonucleotides 299\u003c\/p\u003e \u003cp\u003e11.8.2 CpG and Immunostimulatory (IS) Oligonucleotides 300\u003c\/p\u003e \u003cp\u003e11.8.3 microRNA Mimetics\/Antagonists and siRNAs 301\u003c\/p\u003e \u003cp\u003e11.8.4 Aptamer Oligonucleotides 303\u003c\/p\u003e \u003cp\u003e11.9 Conclusions 304\u003c\/p\u003e \u003cp\u003eAcknowledgments 305\u003c\/p\u003e \u003cp\u003eReferences 305\u003c\/p\u003e \u003cp\u003e\u003cb\u003e12 Specific Considerations for Preclinical Development of Inhaled Oligonucleotides 311\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eNicolay Ferrar \u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e12.1 Background 311\u003c\/p\u003e \u003cp\u003e12.2 Oligonucleotide Delivery Systems 312\u003c\/p\u003e \u003cp\u003e12.2.1 Inhalation Exposure Systems 312\u003c\/p\u003e \u003cp\u003e12.2.2 Intratracheal Aerosol Instillation 313\u003c\/p\u003e \u003cp\u003e12.3 Repeat‐dose Toxicity 314\u003c\/p\u003e \u003cp\u003e12.3.1 General Principles 314\u003c\/p\u003e \u003cp\u003e12.3.2 Recovery Phase 317\u003c\/p\u003e \u003cp\u003e12.4 Toxicokinetics 319\u003c\/p\u003e \u003cp\u003e12.5 Safety Pharmacology 322\u003c\/p\u003e \u003cp\u003e12.5.1 Respiratory System 323\u003c\/p\u003e \u003cp\u003e12.5.2 Cardiovascular and Central Nervous Systems 324\u003c\/p\u003e \u003cp\u003e12.6 Additional Testing 326\u003c\/p\u003e \u003cp\u003e12.6.1 Complement Activation 326\u003c\/p\u003e \u003cp\u003e12.6.2 Proinflammatory Effects 327\u003c\/p\u003e \u003cp\u003e12.7 Conclusion 328\u003c\/p\u003e \u003cp\u003eReferences 328\u003c\/p\u003e \u003cp\u003e\u003cb\u003e13 Lessons Learned in Oncology Programs 331\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eCindy Jacobs, Monica Krieger, Patricia S. Stewart, Karen D. Wisont,and Scott Cormack\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e13.1 Introduction 331\u003c\/p\u003e \u003cp\u003e13.2 Clinical Development of First‐generation ASOs 332\u003c\/p\u003e \u003cp\u003e13.2.1 Aprinocarsen 332\u003c\/p\u003e \u003cp\u003e13.2.2 Oblimersen 334\u003c\/p\u003e \u003cp\u003e13.2.3 Challenges Associated with First‐generation ASOs 335\u003c\/p\u003e \u003cp\u003e13.3 Clinical Development of Second‐generation ASOs 336\u003c\/p\u003e \u003cp\u003e13.3.1 Custirsen 337\u003c\/p\u003e \u003cp\u003e13.3.2 Lessons Learned from Custirsen Clinical Development 343\u003c\/p\u003e \u003cp\u003e13.3.3 Apatorsen 344\u003c\/p\u003e \u003cp\u003e13.3.4 Bladder Cancer 346\u003c\/p\u003e \u003cp\u003e13.3.5 Lung Cancer 346\u003c\/p\u003e \u003cp\u003e13.3.6 Pancreatic Cancer 347\u003c\/p\u003e \u003cp\u003e13.3.7 Prostate Cancer 347\u003c\/p\u003e \u003cp\u003e13.4 Regulatory Considerations 348\u003c\/p\u003e \u003cp\u003e13.5 Future Opportunities for ASOs as Therapeutic Agents for Cancer Treatment 349\u003c\/p\u003e \u003cp\u003eReferences 349\u003c\/p\u003e \u003cp\u003e\u003cb\u003e14 Inhaled Antisense for Treatment of Respiratory Disease 355\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eGail M. Gauvreau, Beth E. Davis, and John Paul Oliveria\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e14.1 Introduction 355\u003c\/p\u003e \u003cp\u003e14.2 Atopic Asthma 355\u003c\/p\u003e \u003cp\u003e14.2.1 Pharmacotherapy of Asthma 356\u003c\/p\u003e \u003cp\u003e14.2.2 Anti‐IL‐5 Monoclonal Antibodies 357\u003c\/p\u003e \u003cp\u003e14.2.3 Anti‐IL‐4\/13 Monoclonal Antibodies 359\u003c\/p\u003e \u003cp\u003e14.3 Antisense Oligonucleotides in Animal Models 361\u003c\/p\u003e \u003cp\u003e14.3.1 CpG Immunostimulatory Sequences 361\u003c\/p\u003e \u003cp\u003e14.3.2 Antisense to Receptors on Eosinophils 366\u003c\/p\u003e \u003cp\u003e14.3.3 Antisense to IL‐4 and IL‐13 Receptors 368\u003c\/p\u003e \u003cp\u003e14.3.4 Summary of Antisense Oligonucleotides in Animal Models 368\u003c\/p\u003e \u003cp\u003e14.4 Clinical Data 369\u003c\/p\u003e \u003cp\u003e14.4.1 Allergen Challenge: A Model of Asthma Exacerbation 369\u003c\/p\u003e \u003cp\u003e14.4.2 Allergen Challenge for Evaluation of Efficacy 369\u003c\/p\u003e \u003cp\u003e14.4.3 1018 Immunostimulatory Sequence 370\u003c\/p\u003e \u003cp\u003e14.4.3.1 Study Design for 1018 ISS 370\u003c\/p\u003e \u003cp\u003e14.4.3.2 Results for 1018 ISS 371\u003c\/p\u003e \u003cp\u003e14.4.4 AIR645 372\u003c\/p\u003e \u003cp\u003e14.4.4.1 Study Design for AIR645 373\u003c\/p\u003e \u003cp\u003e14.4.4.2 Results for AIR645 373\u003c\/p\u003e \u003cp\u003e14.4.5 TPI ASM8 374\u003c\/p\u003e \u003cp\u003e14.4.5.1 Mechanism of TPI ASM8 374\u003c\/p\u003e \u003cp\u003e14.4.5.2 Study #1 for TPI ASM8 375\u003c\/p\u003e \u003cp\u003e14.4.5.3 Study #2 for TPI ASM8 377\u003c\/p\u003e \u003cp\u003e14.5 General\u003c\/p\u003e \u003cp\u003eConclusion 378\u003c\/p\u003e \u003cp\u003eReferences 378\u003c\/p\u003e \u003cp\u003e\u003cb\u003e15 Antisense Oligonucleotides for Treatment of Neurological Diseases 389\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eRosanne Seguin\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e15.1 Introduction 389\u003c\/p\u003e \u003cp\u003e15.1.1 Delivery of ASO to Central Nervous System 389\u003c\/p\u003e \u003cp\u003e15.2 Potential ASO Therapies in Neurodegenerative Diseases 390\u003c\/p\u003e \u003cp\u003e15.2.1 Spinal Muscular Atrophy (SMA) 390\u003c\/p\u003e \u003cp\u003e15.2.2 Amyotrophic Lateral Sclerosis (ALS) 393\u003c\/p\u003e \u003cp\u003e15.2.3 Huntington’s Disease (HD) 396\u003c\/p\u003e \u003cp\u003e15.2.4 Muscular Sclerosis (MS) 399\u003c\/p\u003e \u003cp\u003e15.2.5 Alzheimer’s Disease (AD) 401\u003c\/p\u003e \u003cp\u003e15.3 Conclusion 403\u003c\/p\u003e \u003cp\u003eReferences 403\u003c\/p\u003e \u003cp\u003e\u003cb\u003e16 Nucleic Acids as Adjuvants 411\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eKevin Brown, Montserrat Puig, Lydia Haile, Derek Ireland, John Martucci, and Daniela Verthelyi\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e16.1 Introduction 411\u003c\/p\u003e \u003cp\u003e16.1.1 TLR as Nucleic Acid‐Sensing Pathogen Recognition Receptors (PRR) 412\u003c\/p\u003e \u003cp\u003e16.2 Categories of Nucleic Acid Adjuvants 413\u003c\/p\u003e \u003cp\u003e16.2.1 DNA‐Based Adjuvants and Vaccine Studies in Mice 417\u003c\/p\u003e \u003cp\u003e16.2.2 Classes of CpG ODN that Activate Human TLR9 421\u003c\/p\u003e \u003cp\u003e16.2.3 Preclinical Studies with Human CpG ODN 422\u003c\/p\u003e \u003cp\u003e16.2.4 Safety Issues Raised in Animal Models 424\u003c\/p\u003e \u003cp\u003e16.2.5 Clinical Trial Experience 425\u003c\/p\u003e \u003cp\u003e16.2.6 Safety Issues from Human Clinical Trials 427\u003c\/p\u003e \u003cp\u003e16.2.7 Novel Delivery Systems for CpG ODN as Adjuvants 427\u003c\/p\u003e \u003cp\u003e16.3 Conclusion 429\u003c\/p\u003e \u003cp\u003eAcknowledgments 429\u003c\/p\u003e \u003cp\u003eReferences 430\u003c\/p\u003e \u003cp\u003e\u003cb\u003e17 Splice‐Switching Oligonucleotides 445\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eIsabella Gazzoli and Annemieke Aartsma‐Rus\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e17.1 Introduction of Splice Switching 445\u003c\/p\u003e \u003cp\u003e17.1.1 Correct Cryptic Splicing 446\u003c\/p\u003e \u003cp\u003e17.1.1.1 β‐Thalassemia 446\u003c\/p\u003e \u003cp\u003e17.1.1.2 Cystic Fibrosis 450\u003c\/p\u003e \u003cp\u003e17.1.2 Isoform Switching 451\u003c\/p\u003e \u003cp\u003e17.1.2.1 Anticancer 451\u003c\/p\u003e \u003cp\u003e17.1.2.2 Tauopathies 452\u003c\/p\u003e \u003cp\u003e17.1.3 Induce Exon Inclusion 452\u003c\/p\u003e \u003cp\u003e17.1.3.1 Tumorigenesis 452\u003c\/p\u003e \u003cp\u003e17.1.3.2 Spinal Muscular Atrophy (SMA) 453\u003c\/p\u003e \u003cp\u003e17.1.4 Reading Frame Correction 454\u003c\/p\u003e \u003cp\u003e17.1.4.1 Duchenne Muscular Dystrophy 454\u003c\/p\u003e \u003cp\u003e17.1.4.2 Dysferlinopathies 455\u003c\/p\u003e \u003cp\u003e17.1.5 Knockdown 456\u003c\/p\u003e \u003cp\u003e17.1.5.1 Atherosclerosis 456\u003c\/p\u003e \u003cp\u003e17.1.5.2 Myostatin‐Related Muscle Hypertrophy 457\u003c\/p\u003e \u003cp\u003e17.2 Preclinical and Clinical Development of Splice‐switching Oligos 457\u003c\/p\u003e \u003cp\u003e17.2.1 Introduction to Different Chemistries to be Used for Splice Switching 457\u003c\/p\u003e \u003cp\u003e17.2.2 AON Targets 459\u003c\/p\u003e \u003cp\u003e17.2.3 AON Development for DMD 460\u003c\/p\u003e \u003cp\u003e17.2.4 2′‐O‐Methyl Phosphorothioate AONs 461\u003c\/p\u003e \u003cp\u003e17.2.4.1 Animal Studies 461\u003c\/p\u003e \u003cp\u003e17.2.4.2 Human Studies 463\u003c\/p\u003e \u003cp\u003e17.2.5 Phosphorodiamidate Morpholino Oligos 466\u003c\/p\u003e \u003cp\u003e17.2.5.1 Animal Studies 466\u003c\/p\u003e \u003cp\u003e17.2.5.2 Human Studies 467\u003c\/p\u003e \u003cp\u003e17.2.6 Other Chemistries 468\u003c\/p\u003e \u003cp\u003e17.2.6.1 Peptide‐Conjugated PMOs 468\u003c\/p\u003e \u003cp\u003e17.2.7 Preclinical and Clinical Studies for Other Diseases 470\u003c\/p\u003e \u003cp\u003e17.2.7.1 Spinal Muscular Atrophy (SMA) 470\u003c\/p\u003e \u003cp\u003e17.2.8 Biomarkers 472\u003c\/p\u003e \u003cp\u003e17.3 Future Directions 474\u003c\/p\u003e \u003cp\u003eConflictof Interest 475\u003c\/p\u003e \u003cp\u003eAcknowledgments 475\u003c\/p\u003e \u003cp\u003eReferences 475\u003c\/p\u003e \u003cp\u003e\u003cb\u003e18 CMC Aspects for the Clinical Development of Spiegelmers 491\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003ci\u003eStefan Vonhoff\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e18.1 Introduction 491\u003c\/p\u003e \u003cp\u003e18.2 Technology (Mirror‐imaged SELEX Process) Selected Pharmaceutical Properties 492\u003c\/p\u003e \u003cp\u003e18.3 Preclinical Efficacy Data for Spiegelmers 494\u003c\/p\u003e \u003cp\u003e18.4 Clinical Development 504\u003c\/p\u003e \u003cp\u003e18.4.1 Emapticap Pegol: NOX‐E36 504\u003c\/p\u003e \u003cp\u003e18.4.2 Olaptesed Pegol: NOX‐A12 506\u003c\/p\u003e \u003cp\u003e18.4.3 Lexaptepid Pegol: NOX‐H94 507\u003c\/p\u003e \u003cp\u003e18.5 CMC Aspects for the Development of Spiegelmers 508\u003c\/p\u003e \u003cp\u003e18.5.1 Discovery and Early Preclinical Stage 508\u003c\/p\u003e \u003cp\u003e18.5.2 Generic Manufacturing Process 509\u003c\/p\u003e \u003cp\u003e18.5.2.1 Solid‐phase Synthesis 510\u003c\/p\u003e \u003cp\u003e18.5.2.2 Deprotection 510\u003c\/p\u003e \u003cp\u003e18.5.2.3 Purification of the Intermediate Spiegelmer Prior to Pegylation 510\u003c\/p\u003e \u003cp\u003e18.5.2.4 Pegylation 510\u003c\/p\u003e \u003cp\u003e18.5.2.5 Purification of the Pegylated Spiegelmer 510\u003c\/p\u003e \u003cp\u003e18.5.3 CMC Aspects for the Selection of Development Candidates 511\u003c\/p\u003e \u003cp\u003e18.5.4 GMP Production of Spiegelmers 514\u003c\/p\u003e \u003cp\u003e18.5.4.1 Starting Materials 514\u003c\/p\u003e \u003cp\u003e18.5.4.2 Drug Substance 516\u003c\/p\u003e \u003cp\u003e18.5.4.3 Drug Product 516\u003c\/p\u003e \u003cp\u003e18.5.5 Analytical Methods for the Quality Control of Spiegelmers 517\u003c\/p\u003e \u003cp\u003e18.6 Future Prospects for Spiegelmer Therapeutics 521\u003c\/p\u003e \u003cp\u003eReferences 521\u003c\/p\u003e \u003cp\u003eIndex 527\u003c\/p\u003e   \u003cp\u003e\u003cb\u003eNicolay Ferrari, PhD,\u003c\/b\u003e is the Executive Director of the Canadian Critical Care Trials Group, a Canadian investigator-lead research network, Quebec, Canada. A former Director of Research in Pharmacology at Topigen Pharmaceuticals, Inc, over twenty years of research experience, Dr. Ferrari is the co-inventor of six patents. \u003c\/p\u003e\u003cp\u003e\u003cb\u003eRosanne Seguin,\u003c\/b\u003e \u003cb\u003ePhD,\u003c\/b\u003e is an Academic Associate at the Montreal Neurological Institute of McGill University, Montreal, Quebec, Canada. A former Director of Immunology and Development Support at Topigen Pharmaceuticals, Inc. Dr. Seguin has 20 years of research experience.    \u003c\/p\u003e\u003cp\u003e\u003cb\u003eA COMPREHENSIVE REVIEW OF CONTEMPORARY ANTISENSE OLIGONUCLEOTIDES DRUGS AND THERAPEUTIC PRINCIPLES, METHODS, APPLICATIONS, AND RESEARCH\u003c\/b\u003e \u003c\/p\u003e\u003cp\u003eOligonucleotide-based drugs, in particular antisense oligonucleotides, are part of a growing number of pharmaceutical and biotech programs progressing to treat a wide range of indications including cancer, cardiovascular, neurodegenerative, neuromuscular, and respiratory diseases, as well as other severe and rare diseases. Reviewing fundamentals and offering guidelines for drug discovery and development, this book is a practical guide covering all key aspects of this increasingly popular area of pharmacology and biotech and pharma research, from the basic science behind antisense oligonucleotides chemistry, toxicology, manufacturing, to safety assessments, the design of therapeutic protocols, to clinical experience. \u003c\/p\u003e\u003cp\u003eAntisense oligonucleotides are single strands of DNA or RNA that are complementary to a chosen sequence. While the idea of antisense oligonucleotides to target single genes dates back to the 1970's, most advances have taken place in recent years. The increasing number of antisense oligonucleotide programs in clinical development is a testament to the progress and understanding of pharmacologic, pharmacokinetic, and toxicologic properties as well as improvement in the delivery of oligonucleotides. This valuable book reviews the fundamentals of oligonucleotides, with a focus on antisense oligonucleotide drugs, and reports on the latest research underway worldwide. \u003c\/p\u003e\u003cul\u003e \u003cli\u003eHelps readers understand antisense molecules and their targets, biochemistry, and toxicity mechanisms, roles in disease, and applications for safety and therapeutics\u003c\/li\u003e \u003cli\u003eExamines the principles, practices, and tools for scientists in both pre-clinical and clinical settings and how to apply them to antisense oligonucleotides\u003c\/li\u003e \u003cli\u003eProvides guidelines for scientists in drug design and discovery to help improve efficiency, assessment, and the success of drug candidates\u003c\/li\u003e \u003cli\u003eIncludes interdisciplinary perspectives, from academia, industry, regulatory and from the fields of pharmacology, toxicology, biology, and medicinal chemistry\u003c\/li\u003e \u003c\/ul\u003e \u003cp\u003e\u003ci\u003eOligonucleotide-Based Drugs and Therapeutics\u003c\/i\u003e belongs on the reference shelves of chemists, pharmaceutical scientists, chemical biologists, toxicologists and other scientists working in the pharmaceutical and biotechnology industries. It will also be a valuable resource for regulatory specialists and safety assessment professionals and an important reference for academic researchers and post-graduates interested in therapeutics, antisense therapy, and oligonucleotides.\u003c\/p\u003e","brand":"Wiley","offers":[{"title":"Default Title","offer_id":47989711864037,"sku":"NP9781118537336","price":231.95,"currency_code":"USD","in_stock":false}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1842\/7735\/files\/9781118537336.jpg?v=1761785205","url":"https:\/\/k12savings.com\/products\/oligonucleotide-based-drugs-and-therapeutics-isbn-9781118537336","provider":"K12savings","version":"1.0","type":"link"}