ISBN: 3540659277
TITLE: Novel Therapeutics from Modern Biotechnology
AUTHOR: Oxender, Dale L.; Post, Leonard E. (Eds.)
TOC:

CHAPTER 1
Overview of Regulatory Expectations for Introducing Novel Therapies into Clinical Trials
J.A. CAVAGNARO and M.A. SERABIAN 1
A. Introduction 1
B. Roles of Regulatory Scientists 2
C. Product Development and Availability 2
D. Data Requirements 4
E. Manufacturing 4
F. Preclinical Safety Testing 5
G. Case-By-Case Approach 6
H. Testing Goals 6
I. Study Design 7
J. Defining Exposure 7
K. Product-Specific Concerns 8
L. Accessibility of Preclinical Safety Data 9
M. Clinical Studies 10
I. Early Development 10
II. Late Development 10
N. Summary 11
References 11
CHAPTER 2
Preparation of Clinical Trial Supplies of Biopharmaceuticals
A.S. LUBINIECKI, J.C. ERICKSON, C.L. NOLAN, R.G. SCOTT, P.J. SHADLE, T.M. SMITH, and D.W. ZABRISKIE. With 1 Figure 13
A. Introduction 13
I. Research Support Systems 13
B. Preclinical Studies 13
C. Clinical Supplies 15
I. Fermentation 15
II. Harvest 17
III. Purification 18
D. Purification of rDNA-Derived Anti-RSV MAb 19
E. Product Quality Issues 22
I. Protein Purity 22
II. Protein Integrity 22
III. Microbial and Viral Safety 23
IV. Other Contaminants 23
F. Process Design and Validation 24
I. Validation of Endotoxin Removal 24
II. Process Validation of Model Virus Clearance 26
G. Process Economics and the Future of Chromatography 27
I. Process Automation and Control 27
II. Generic Purification Methods 30
H. Conclusions 31
References 31
CHAPTER 3
Proteins as Drugs: Analysis, Formulation and Delivery
C.R. MIDDAUGH and R. PEARLMAN 33
A. Introduction 33
B. The Analysis of Protein Pharmaceuticals 34
I. X-Ray Crystallography 36
II. Nuclear Magnetic Resonance 38
III. Mass Spectroscopy 39
IV. Multiple Parametric Approaches 40
V. Miscellaneous Comments 42
C. Formulation 44
D. Delivery 49
I. Controlled-Release Dosage Forms 51
II. A Practical Delivery Challenge: Insulin 52
References 54
CHAPTER 4
Strategies for Dealing With the Immunogenicity of Therapeutic Proteins
M.L. NUCCI, R.G.L. SHORR, and A. ABUCHOWSKI 59
A. Introduction 59
B. Case Histories of Protein Therapeutic Development 61
I. Insulin 61
II. Growth Hormone 63
III. Asparaginase 64
IV. Glucocerebrosidase 65
V. OKT3 66
C. Strategies Under Development for Increasing the Therapeutic Value of Proteins and Peptides 67
I. Encapsulation 67
II. Non-Parenteral Routes of Administration 70
III. Targeting 72
IV. Conjugation 75
V. Protein Engineering 77
D. Choosing the Proper Strategy for a Protein Therapeutic 79
E. The Future of Protein Therapeutics 80
References 80
CHAPTER 5
Targeted Toxin Hybrid Proteins
R.J. KREITMAN and I. PASTAN. With 6 Figures 89
A. Introduction 89
I. Protein Toxins That Inhibit Protein Synthesis 89
1. Plant Toxins 89
2. Bacterial Toxins 89
II. Structure and Function of Pseudomonas Exotoxin 90
1. Definition of Domains and Mechanism of Intoxication 90
2. Mutants Lacking Cell Binding 91
III. Types of Toxins Made with PE 91
1. Chemical Conjugates vs Recombinant Fusions 91
2. Fusion Toxins Containing Transforming Growth Factor-alpha 91
3. Fusion Toxins Containing Interleukin 2 92
4. Recombinant Immunotoxins 93
B. Preclinical Development of Anti-Tac(Fv) Toxins 93
I. Background 93
II. Efficacy Data on Relevant Human Cells 94
1. Human Activated T-Lymphocytes 94
2. Fresh Adult T-Cell Leukemia Cells 95
3. Fresh Chronic Lymphocytic Leukemia Cells 96
III. Efficacy Data in an Animal Model of IL2R-Bearing Cancer 97
1. Production of the Human ATAC4 Line 97
2. Toxicity of Anti-Tac(Fv) Toxins in Mice 97
3. Pharmacokinetics in Mice 97
4. Antitumor Activity in Tumor-Bearing Mice 98
IV. Primate Testing 99
1. Pharmacokinetics in Cynomolgus Monkeys 99
2. Toxicity in Cynomolgus Monkeys 100
V. Production Issues 100
C. Preclinical Development of Inerleukin 6-PE^4E 102
I. Background 102
II. Study of IL6-PE^4E For Ex Vivo Marrow Purging in Multiple Myeloma 102
1. Rationale 102
2. Efficacy Against Fresh Marrow Cells from Myeloma Patients 103
3. Safety Toward Fresh Normal Marrow Cells 103
4. Safety of IL6-PE^4E Toward Normal Hematopoietic Progenitors 104
5. Lack of Prevention of Bone Marrow Engraftment 104
6. Carryover of IL6-PE^4E In Vivo 104
III. Production of IL6-PE^4E 105
D. Summary 105
References 106
CHAPTER 6
SB 209763: A Humanized Monoclonal Antibody for the Prophylaxis and Treatment of Respiratory Syncytial Virus Infection
T.G. PORTER, S.G. GRIEGO, T.K. HART, D.E. EVERITT, and S.B. DILLON. With 4 Figures 111
A. Introduction 111
B. Early Challenges in the Development of SB 209763 112
I. Selection of Target Antigen 112
II. Molecular Engineering of SB 209763 113
III. Production 114
IV. Primary Structure Analysis 115
C. Preclinical Evaluation Prior to Testing in Humans 115
I. Fusion Inhibition: An In Vitro Correlate of Protection 116
II. Antigenic Variation 119
III. Animal Models of Respiratory Syncytial Virus Infection 119
IV. Safety and Pharmacokinetics 120
D. Challenges for the Early Clinical Development of SB 209763 122
I. Selection of the Initial Study Population and Safety Considerations 122
II. Pharmacodynamic Markers to Establish Pharmacologic Effect 124
III. Formulation Considerations for Clinical Studies 124
IV. Surveillance for Anti-SB 209763 Antibodies 124
V. Transition to the Target Pediatric Population and Choice of Dose 125
VI. Results of Early Clinical Studies 125
E. Conclusion 127
References 127
CHAPTER 7
Preclinical Development of Antisense Therapeutics
A.A. LEVIN, S.P. HENRY, C.F. BENNETT, D.L. COLE, G.E. HARDEE, and G.S. SRIVATSA. With 2 Figures 131
A. Introduction 131
B. Pharmacology of Antisense Oligodeoxynucleotides 131
I. Molecular Pharmacology of Antisense Oligodeoxynucleotides 131
II. In Vivo Pharmacology of Antisense Oligodeoxynucleotides 134
C. Pharmacokinetics and Toxicity of Oligodeoxynucleotide Therapeutics 136
I. Pharmacokinetics and Metabolism 136
II. Toxicity of Phosphorothioate Oligodeoxynucleotides 142
D. Chemistry, Manufacture and Control of Phosphorothioate Oligodeoxynucleotide Drugs 147
I. Synthesis of Phosphorothioate Oligodeoxynucleotides 149
1. Chemistry of Elongation 149
2. Chemistry of Sulfurization 150
3. 0,0-Linked Phosphorothioate DNA Diastereoisomerism 151
II. Purification of Phosphorothioate Oligodeoxynucleotides 152
III. Quality Control of Phosphorothioate Oligodeoxynucleotides 153
E. Formulation and Drug Delivery of Oligodeoxynucleotides 154
I. Physical-Chemical Properties 154
II. Formulation 155
III. Drug Delivery: Targeting, Uptake and Release 156
F. Summary 157
References 157
CHAPTER 8
Formulation and Delivery of Nucleic Acids
H.E.J. HOFLAND and L. HUANG. With 6 Figures 165
A. Introduction 165
B. Formulation of DNA 166
I. Naked-DNA Injections 166
II. Gene Guns 167
III. Polymer-Based Formulations 167
IV. Lipid-Based Formulations 171
1. Liposome Encapsulation 171
2. Cationic Lipid/Nucleotide Complex 171
3. DNA-Binding Moiety 172
4. Hydrophobic Moiety 176
5. Spacer 177
6. Linker 177
7. Helper Lipid 178
C. Delivery to Target Cells 180
D. Cell Entry 182
I. Receptor-Mediated Uptake 182
E. Endosomal Release 183
F. Nuclear Localization 185
G. Gene Expression 186
References 187
CHAPTER 9
Safe, Efficient Production of Retroviral Vectors
H. KOTANI and G.J. McGARRITY. With 11 Figures 193
A. Introduction 193
B. Vectors 194
I. Retroviral Vectors 194
C. Production of Retroviral Vectors 196
I. Production Methods 198
1. Batch Systems 198
2. Roller Bottles 198
3. Multilayered Propagator 200
II. Bioreactors 200
1. CellCube Bioreactor 200
2. Hollow-Fiber Bioreactor 201
3. Microcarrier Beads in Bioreactor 202
4. Packed-Bed Air-Lift Bioreactor 202
5. Serum-Containing Production 203
D. Downstream Processing 207
E. GMP Production of Retroviral Vectors 208
I. Cell Banking 209
II. Serum-Free Upstream Processing 211
III. Serum-Free Downstream Processing 211
F. In-Process Assays 212
G. Quality Control 215
H. Safety 215
I. Summary and Conclusions 217
References 218
CHAPTER 10
Clinical Systems for the Production of Cells and Tissues for Human Therapy
R.D. ARMSTRONG, M.R. KOLLER, J. MALUTA, and W.C. OGIER.
With 4 Figures 221
A. Introduction 221
B. Cell Therapy and Tissue Engineering 222
I. Ex Vivo Gene Therapy 223
II. Stem-Cell Therapy 223
C. Critical Requirements for Ex Vivo Cell Production 225
I. Process Reliability and Control: Automation 225
II. Process Sterility: Closed Systems 226
III. Cell Recovery 228
IV. Optimization of Key Culture Parameters by Design 228
V. Good Manufacturing Practices 230
D. Cell-Culture Devices and Procedures 231
I. Traditional Cell-Culture Processes: Research Laboratory Environment 232
1. Culture Flasks and Roller Bottles 233
2. Flexible Tissue Culture Containers 233
3. Bioreactors 234
II. AASTROM Cell-Production System 235
1. System Description 235
a. Disposable Cell Cassette 236
b. Incubator 237
c. Processor 237
d. System Manager 238
e. ID Key 238
E. Applications for On-Site Delivery of Therapeutic Cell Production 238
I. Bone-Marrow Cell Production 239
II. Other Cell and Tissue Production 239
F. Summary 240
References 240
Subject Index 243
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