Novel Therapeutics from Modern Biotechnology

1 Overview of Regulatory Expectations for Introducing Novel Therapies into Clinical Trials.- A. Introduction.- B. Roles of Regulatory Scientists.- C. Product Development and Availability.- D. Data Requirements.- E. Manufacturing.- F. Preclinical Safety Testing.- G. Case-By-Case Approach.- H. Testing Goals.- I. Study Design.- J. Defining Exposure.- K. Product-Specific Concerns.- L. Accessibility of Preclinical Safety Data.- M. Clinical Studies.- I. Early Development.- II. Late Development.- N. Summary.- References.- 2 Preparation of Clinical Trial Supplies of Biopharmaceuticals.- A. Introduction.- I. Research Support Systems.- B. Preclinical Studies.- C. Clinical Supplies.- I. Fermentation.- II. Harvest.- III. Purification.- D. Purification of rDNA-Derived Anti-RSV MAb.- E. Product Quality Issues.- I. Protein Purity.- II. Protein Integrity.- III. Microbial and Viral Safety.- IV. Other Contaminants.- F. Process Design and Validation.- I. Validation of Endotoxin Removal.- II. Process Validation of Model Virus Clearance.- G. Process Economics and the Future of Chromatography.- I. Process Automation and Control.- II. Generic Purification Methods.- H. Conclusions.- References.- 3 Proteins as Drugs: Analysis, Formulation and Delivery.- A. Introduction.- B. The Analysis of Protein Pharmaceuticals.- I. X-Ray Crystallography.- II. Nuclear Magnetic Resonance.- III. Mass Spectroscopy.- IV. Multiple Parametric Approaches.- V. Miscellaneous Comments.- C. Formulation.- D. Delivery.- I. Controlled-Release Dosage Forms.- II. A Practical Delivery Challenge: Insulin.- References.- 4 Strategies for Dealing With the Immunogenicity of Therapeutic Proteins.- A. Introduction.- B. Case Histories of Protein Therapeutic Development.- I. Insulin.- II. Growth Hormone.- III. Asparaginase.- IV. Glucocerebrosidase.- V. OKT3.- C. Strategies Under Development for Increasing the Therapeutic Value of Proteins and Peptides.- I. Encapsulation.- II. Non-Parenteral Routes of Administration.- III. Targeting.- IV. Conjugation.- V. Protein Engineering.- D. Choosing the Proper Strategy for a Protein Therapeutic.- E. The Future of Protein Therapeutics.- References.- 5 Targeted Toxin Hybrid Proteins.- A. Introduction.- I. Protein Toxins That Inhibit Protein Synthesis.- 1. Plant Toxins.- 2. Bacterial Toxins.- II. Structure and Function of Pseudomonas Exotoxin.- 1. Definition of Domains and Mechanism of Intoxication.- 2. Mutants Lacking Cell Binding.- III. Types of Toxins Made with PE.- 1. Chemical Conjugates vs Recombinant Fusions.- 2. Fusion Toxins Containing Transforming Growth Factor-?.- 3. Fusion Toxins Containing Interleukin 2.- 4. Recombinant Immunotoxins.- B. Preclinical Development of Anti-Tac(Fv) Toxins.- I. Background.- II. Efficacy Data on Relevant Human Cells.- 1. Human Activated T-Lymphocytes.- 2. Fresh Adult T-Cell Leukemia Cells.- 3. Fresh Chronic Lymphocytic Leukemia Cells.- III. Efficacy Data in an Animal Model of IL2R-Bearing Cancer.- 1. Production of the Human ATAC-4 Line.- 2. Toxicity of Anti-Tac(Fv) Toxins in Mice.- 3. Pharmacokinetics in Mice.- 4. Antitumor Activity in Tumor-Bearing Mice.- IV. Primate Testing.- 1. Pharmacokinetics in Cynomolgus Monkeys.- 2. Toxicity in Cynomolgus Monkeys.- V. Production Issues.- C. Preclinical Development of Inerleukin 6-PE4E.- I. Background.- II. Study of IL6-PE4E For Ex Vivo Marrow Purging in Multiple Myeloma.- 1. Rationale.- 2. Efficacy Against Fresh Marrow Cells from Myeloma Patients.- 3. Safety Toward Fresh Normal Marrow Cells.- 4. Safety of IL6-PE4EToward Normal Hematopoietic Progenitors.- 5. Lack of Prevention of Bone Marrow Engraftment.- 6. Carryover of IL6-PE4E In Vivo.- III. Production of IL6-PE4E.- D. Summary.- References.- 6 SB 209763: A Humanized Monoclonal Antibody for the Prophylaxis and Treatment of Respiratory Syncytial Virus Infection.- A. Introduction.- B. Early Challenges in the Development of SB 209763.- I. Selection of Target Antigen.- II. Molecular Engineering of SB 209763.- III. Production.- IV. Primary Structure Analysis.- C. Preclinical Evaluation Prior to Testing in Humans.- I. Fusion Inhibition: An In Vitro Correlate of Protection.- II. Antigenic Variation.- III. Animal Models of Respiratory Syncytial Virus Infection.- IV. Safety and Pharmacokinetics.- D. Challenges for the Early Clinical Development of SB 209763.- I. Selection of the Initial Study Population and Safety Considerations.- II. Pharmacodynamic Markers to Establish Pharmacologic Effect.- III. Formulation Considerations for Clinical Studies.- IV. Surveillance for Anti-SB 209763 Antibodies.- V. Transition to the Target Pediatric Population and Choice of Dose.- VI. Results of Early Clinical Studies.- E. Conclusion.- References.- 7 Preclinical Development of Antisense Therapeutics.- A. Introduction.- B. Pharmacology of Antisense Oligodeoxynucleotides.- I. Molecular Pharmacology of Antisense Oligodeoxynucleotides.- II. In Vivo Pharmacology of Antisense Oligodeoxynucleotides.- C. Pharmacokinetics and Toxicity of Oligodeoxynucleotide Therapeutics.- I. Pharmacokinetics and Metabolism.- II. Toxicity of Phosphorothioate Oligodeoxynucleotides.- D. Chemistry, Manufacture and Control of Phosphorothioate Oligodeoxynucleotide Drugs.- I. Synthesis of Phosphorothioate Oligodeoxynucleotides.- 1. Chemistry of Elongation.- 2. Chemistry of Sulfurization.- 3. O,O-Linked Phosphorothioate DNA Diastereoisomerism.- II. Purification of Phosphorothioate Oligodeoxynucleotides.- III. Quality Control of Phosphorothioate Oligodeoxynucleotides.- E. Formulation and Drug Delivery of Oligodeoxynucleotides.- I. Physical—Chemical Properties.- II. Formulation.- III. Drug Delivery: Targeting, Uptake and Release.- F. Summary.- References.- 8 Formulation and Delivery of Nucleic Acids.- A. Introduction.- B. Formulation of DNA.- I. Naked-DNA Injections.- II. Gene Guns.- III. Polymer-Based Formulations.- IV. Lipid-Based Formulations.- 1. Liposome Encapsulation.- 2. Cationic Lipid/Nucleotide Complex.- 3. DNA-Binding Moiety.- 4. Hydrophobic Moiety.- 5. Spacer.- 6. Linker.- 7. Helper Lipid.- C. Delivery to Target Cells.- D. Cell Entry.- I. Receptor-Mediated Uptake.- E. Endosomal Release.- F. Nuclear Localization.- G. Gene Expression.- References.- 9 Safe, Efficient Production of Retroviral Vectors.- A. Introduction.- B. Vectors.- I. Retroviral Vectors.- C. Production of Retroviral Vectors.- I. Production Methods.- 1. Batch Systems.- 2. Roller Bottles.- 3. Multilayered Propagator.- II. Bioreactors.- 1. CellCube Bioreactor.- 2. Hollow-Fiber Bioreactor.- 3. Microcarrier Beads in Bioreactor.- 4. Packed-Bed Air-Lift Bioreactor.- 5. Serum-Containing Production.- D. Downstream Processing.- E. GMP Production of Retroviral Vectors.- I. Cell Banking.- II. Serum-Free Upstream Processing.- III. Serum-Free Downstream Processing.- F. In-Process Assays.- G. Quality Control.- H. Safety.- I. Summary and Conclusions.- References.- 10 Clinical Systems for the Production of Cells and Tissues for Human Therapy.- A. Introduction.- B. Cell Therapy and Tissue Engineering.- I. Ex Vivo Gene Therapy.- II. Stem-Cell Therapy.- C. Critical Requirements for Ex Vivo Cell Production.- I. Process Reliability and Control: Automation.- II. Process Sterility: Closed Systems.- III. Cell Recovery.- IV. Optimization of Key Culture Parameters by Design.- V. Good Manufacturing Practices.- D. Cell-Culture Devices and Procedures.- I. Traditional Cell-Culture Processes: Research Laboratory Environment.- 1. Culture Flasks and Roller Bottles.- 2. Flexible Tissue Culture Containers.- 3. Bioreactors.- II. AASTROM Cell-Production System.- 1. System Description.- a. Disposable Cell Cassette.- b. Incubator.- c. Processor.- d. System Manager.- e. ID Key.- E. Applications for On-Site Delivery of Therapeutic Cell Production.- I. Bone-Marrow Cell Production.- II. Other Cell and Tissue Production.- F. Summary.- References.