Linker p. 177 Helper Lipid p. 178 Delivery to Target Cells p. 180 Cell Entry p. 182 Receptor-Mediated Uptake p. 182 Endosomai Release p.

Transcription

1 Overview of Regulatory Expectations for Introducing Novel Therapies into Clinical Trials Introduction p. 1 Roles of Regulatory Scientists p. 2 Product Development and Availability p. 2 Data Requirements p. 4 Manufacturing p. 4 Preclinical Safety Testing p. 5 Case-By-Case Approach p. 6 Testing Goals p. 6 Study Design p. 7 Denning Exposure Product-Specific Concerns p. 8 Accessibility of Preclinical Safety Data p. 9 Clinical Studies p. 10 Early Development p. 10 Late Development p. 10 Summary p. 11 References p. 11 Preparation of Clinical Trial Supplies of Biopharmaceutkals p. 13 Introduction p. 13 Research Support Systems p. 13 Preclinical Studies p. 13 Clinical Supplies p. 15 Fermentation p. 15 Harvest p. 17 Purification p. 18 Purification of rdna-derivcd Anti-RSV MAb p. 19 Product Quality Issues p. 22 Protein Purity p. 22 Protein Integrity p. 22 Microbial and Viral Safety p. 23 Other Contaminants p. 23 Process Design and Validation p. 24 Validation of Endotoxin Removal p. 24 Process Validation of Model Virus Clearance p. 26 Process Economics and the Future of Chromatography p. 27 Process Automation and Control p. 27 Generic Purification Methods p. 30 Conclusions p. 31 References p. 31 Proteins as Drugs: Analysis, Formulation and Delivery p. 33 p. 1

2 Introduction p. 33 The Analysis of Protein Pharmaceuticals p. 34 X-Ray Crystallography p. 36 Nuclear Magnetic Resonance p. 38 Mass Spectroscopy p. 39 Multiple Parametric Approaches p. 40 Miscellaneous Comments p. 42 Formulation p. 44 Delivery p. 49 Controlled-Release Dosage Forms p. 51 A Practical Delivery Challenge: Insulin p. 52 References p. 54 Strategies for Dealing With the Immunogenicity of Therapeutic Proteins p. 59 Introduction p. 59 Case Histories of Protein Therapeutic Development p. 61 Insulin p. 61 Growth Hormone p. 63 Asparaginase p. 64 Glucocerebrosidase p. 65 OKT3 p. 66 Strategies Under Development for Increasing the Therapeutic Value of Proteins and Peptides p. 67 Encapsulation p. 67 Non-Parenteral Routes of Administration p. 70 Targeting p. 72 Conjugation p. 75 Protein Engineering & p. 77 Choosing the Proper Strategy for a Protein Therapeutic p. 79 The Future of Protein Therapeutics p. 80 References p. 80 Targeted Toxin Hybrid Proteins p. 89 Introduction p. 89 Protein Toxins That Inhibit Protein Synthesis p. 89 Plant Toxins p. 89 Bacterial Toxins p. 89 Structure and Function of Pseudomonas Exotoxin p. 90 Definition of Domains and Mechanism of Intoxication p. 90 Mutants Lacking Cell Binding p. 91 Types of Toxins Made with PE p. 91 Chemical Conjugates vs Recombinant Fusions p. 91 Fusion Toxins Containing Transforming Growth Factor- p. 91 Fusion Toxins Containing Interleukin 2 p. 92

3 Recombinant Immunoloxins p. 93 Preclinical Development of Anti-Tac(Fv) Toxins p. 93 Background p. 93 Efficacy Data on Relevant Human Cells p. 94 Human Activated T-Lymphocytes p. 94 Fresh Adult T-Cell Leukemia Cells p. 95 Fresh Chronic Lymphocytic Leukemia Cells p. 96 Efficacy Data in an Animal Model of IL2R-Bearing Cancer p. 97 Production of the Human ATAC 4 Line p. 97 Toxicity of Anti-Tac(Ev) Toxins in Mice p. 97 Pharmacokinetics in Mice p. 97 Antitumor Activity in Tumor-Bearing Mice p. 98 Primate Testing p. 99 Pharmacokinetics in Cynomolgus Monkeys p. 99 Toxicity in Cynomolgus Monkeys p. 100 Production Issues p. 100 Preclinical Development of Inerleukin 6-PE p. 102 Background p. 102 Study of IL6-PE p. 102 Rationale p. 102 Efficacy Against Fresh Marrow Cells from Myeloma Patients p. 103 Safety Toward Fresh Normal Marrow Cells p. 103 Safety of IL6-PE p. 104 Lack of Prevention of Bone Marrow Engraftment p. 104 Carryover of IL6-PE p. 104 Production of IL6-PE p. 105 Summary p. 105 References p. 106 SB : A Humanized Monoclonal Antibody for the Prophylaxis and Treatment of Respiratory Syncytial Virus Infection p. 111 Introduction p. 111 Early Challenges in the Development of SB p. 112 Selection of Target Antigen p. 112 Molecular Engineering of SB p. 113 Production p. 114 Primary Structure Analysis p. 115 Preclinical Evaluation Prior to Testing in Humans p. 115 Fusion Inhibition: An In Vitro Correlate of Protection p. 116 Antigenic Variation p. 119 Animal Models of Respiratory Syncytial Virus Infection p. 119 Safely and Pharmacokinetics p. 120 Challenges for the Early Clinical Development of SB p. 122

4 Selection of the Initial Study Population and Safety Considerations p. 122 Pharmacodynamic Markers to Establish Pharmacologic Effect p. 124 Formulation Considerations for Clinical Studies p. 124 Surveillance for Anti-SB Antibodies p. 124 Transition to the Target Pediatric Population and Choice of Dose p. 125 Results of Early Clinical Studies p. 125 Conclusion p. 127 References p. 127 Preclinical Development of Antisense Therapeutics p. 131 Introduction p. 131 Pharmacology of Antisense Oligodeoxynuclcotides p. 131 Molecular Pharmacology of Antisense Oligodeoxynucleotides p. 131 In Vivo Pharmacology of Antisense Oligodeoxynucleotides p. 134 Pharmacokinetics and Toxicity of Oligodeoxynuclcotidc Therapeutics p. 136 Pharmacokinetics and Metabolism p. 136 Toxicity of Phosphorothioate Oligodeoxynucleotides p. 142 Chemistry, Manufacture and Control of Phosphorothioate Oligodcoxynucleotide Drugs p. 147 Synthesis of Phosphorothioate Oligodeoxynucleotides p. 149 Chemistry of Elongation p. 149 Chemistry of Sulfurization p. 150 O,O-Linked Phosphorothioate DNA Diastereoisomerism p. 151 Purification of Phosphorothioate Oligodeoxynucleotides p. 152 Quality Control of Phosphorothioate Oligodeoxynucleotides p. 153 Formulation and Drug Delivery of Oligodeoxynucleotides p. 154 Physical-Chemical Properties p. 154 Formulation p. 155 Drug Delivery: Targeting, Uptake and Release p. 156 Summary p. 157 References p. 157 Formulation and Delivery of Nucleic Acids p. 165 Introduction p. 165 Formulation of DNA p. 166 Nakcd-DNA Injections p. 166 Gene Guns p. 167 Polymer-Based Formulations p. 167 Lipid-Based Formulations p. 171 Liposome Encapsulation p. 171 Cationic Lipid/Nucleotide Complex p. 171 DNA-Binding Moiety p. 172 Hydrophobic Moiety p. 176 Spacer p. 177

5 Linker p. 177 Helper Lipid p. 178 Delivery to Target Cells p. 180 Cell Entry p. 182 Receptor-Mediated Uptake p. 182 Endosomai Release p. 183 Nuclear Localization p. 185 Gene Expression p. 186 References p. 187 Safe, Efficient Produetiun of Retroviral Vectors p. 193 Introduction p. 193 Vectors p. 194 Retroviral Vectors p. 194 Production of Retroviral Vectors p. 196 Production Methods p. 198 Batch Systems p. 198 Roller Bottles p. 198 Mullilayered Propagator p. 200 Bioreactors p. 200 CellCube Bioreactor p. 200 Hollow-Fiber Bioreactor p. 201 Microcarrier Beads in Bioreactor p. 202 Packed-Bed Air-Lift Bioreactor p. 202 Serum-Containing Production p. 203 Downstream Processing p. 207 GMP Production of Retroviral Vectors p. 208 Cell Banking p. 209 Serum-Free Upstream Processing p. 211 Serum-Free Downstream Processing p. 211 In-Process Assays p. 212 Ouality Control p. 215 Safety p. 215 Summary and Conclusions p. 217 References p. 218 Clinical Systems for the Production of Cells and Tissues for Human Therapy p. 221 Introduction p. 221 Cell Therapy and Tissue Engineering p. 222 Ex Vivo Gene Therapy p. 223 Stem-Cell Therapy p. 223 Critical Requirements for Ex Vivo Cell Production p. 225 Process Reliability and Control: Automation p. 225

6 Process Sterility: Closed Systems p. 226 Cell Recovery p. 228 Optimization of Key Culture Parameters by Design p. 228 Good Manufacturing Practices p. 230 Cell-Culture Devices and Procedures p. 231 Traditional Cell-Culture Processes: Research Laboratory Environment p. 232 Culture Flasks and Roller Bottles p. 233 Flexible Tissue Culture Containers p. 233 Bioreaetors p. 234 AASTROM Cell-Production System p. 235 System Description p. 235 Disposable Cell Cassette p. 236 Incubator p. 237 Processor p. 237 System Manager p. 238 ID Key p. 238 Applications for On-Sile Delivery of Therapeutic Cell Production p. 238 Bone-Marrow Cell Production p. 239 Other Cell and Tissue Production p. 239 Summary p. 240 References p. 240 Subject Index p. 243 Table of Contents provided by Blackwell's Book Services and R.R. Bowker. Used with permission.