Preface xvii
Chapter 1 Pharmaceuticals, biologics and biopharmaceuticals 1
Introduction to pharmaceutical products 1
Biopharmaceuticals and pharmaceutical biotechnology 1
History of the pharmaceutical industry 3
The age of biopharmaceuticals 5
Biopharmaceuticals: current status and future prospects 8
Traditional pharmaceuticals of biological origin 12
Pharmaceuticals of animal origin 13
The sex hormones 14
The androgens 14
Oestrogens 15
Progesterone and progestogens 17
Corticosteroids 19
Catecholamines 21
Prostaglandins 23
Pharmaceutical substances of plant origin 27
Alkaloids 28
Atropine and scopalamine 28
Morphine and cocaine 29
Additional plant alkaloids 30
Ergot alkaloids 30
Flavonoids, xanthines and terpenoids 30
Cardiac glycosides and coumarins 33
Aspirin 33
Pharmaceutical substances of microbial origin 33
The macrolides and ansamycins 38
Peptide and other antibiotics 39
Conclusion 39
Further reading 40
Chapter 2 The drug development process 43
Drug discovery 44
The impact of genomics and related technologies upon drug discovery 45
Gene chips 47
Proteomics 49
Structural genomics 50
Pharmacogenetics 51
Plants as a source of drugs 52
Microbial drugs 53
Rational drug design 54
Combinatorial approaches to drug discovery 56
Initial product characterization 57
Patenting 57
What is a patent and what is patentable? 57
Patent types 62
The patent application 63
Patenting in biotechnology 64
Delivery of biopharmaceuticals 66
Oral delivery systems 66
Pulmonary delivery 67
Nasal, transmucosal and transdermal delivery systems 68
Pre-clinical trials 69
Pharmacokinetics and pharmacodynamics 69
Toxicity studies 71
Reproductive toxicity and teratogenicity 71
Mutagenicity, carcinogenicity and other tests 72
Clinical trials 73
Clinical trial design 75
Trial size and study population 75
Randomized control studies 76
Additional trial designs 76
The role and remit of regulatory authorities 78
The Food and Drug Administration 78
The investigational new drug application 80
The new drug application 82
European regulations 84
National regulatory authorities 84
The EMEA and the new EU drug approval systems 85
The centralized procedure 86
Mutual recognition 88
Drug registration in Japan 88
World harmonization of drug approvals 89
Conclusion 89
Further reading 89
Chapter 3 The drug manufacturing process 93
International pharmacopoeia 93
Martindale, the Extra Pharmacopoeia 94
Guides to good manufacturing practice 94
The manufacturing facility 97
Clean rooms 98
Cleaning, decontamination and sanitation (CDS) 101
CDS of the general manufacturing area 102
CDS of process equipment 102
Water for biopharmaceutical processing 104
Generation of purified water and water for injections (WFI) 105
Distribution system for WFI 107
Documentation 109
Specifications 110
Manufacturing formulae, processing and packaging instructions 110
Records 111
Generation of manufacturing records 111
Sources of biopharmaceuticals 112
E. coli as a source of recombinant, therapeutic proteins 112
Expression of recombinant proteins in animal cell culture systems 116
Additional production systems: yeasts 116
Fungal production systems 117
Transgenic animals 118
Transgenic plants 122
Insect cell-based systems 123
Production of final product 124
Cell banking systems 127
Upstream processing 128
Microbial cell fermentation 129
Mammalian cell culture systems 133
Downstream processing 134
Final product formulation 140
Some influences that can alter the biological activity of proteins 142
Proteolytic degradation 143
Protein deamidation 144
Oxidation and disulphide exchange 145
Alteration of glycoprotein glycosylation patterns 147
Stabilizing excipients used in final product formulations 150
Final product fill 153
Freeze-drying 155
Labelling and packing 158
Analysis of the final product 159
Protein-based contaminants 159
Removal of altered forms of the protein of interest from the product stream 160
Product potency 161
Determination of protein concentration 163
Detection of protein-based product impurities 164
Capillary electrophoresis 166
High-pressure liquid chromatography (HPLC) 167
Mass spectrometry 168
Immunological approaches to detection of contaminants 168
Amino acid analysis 169
Peptide mapping 170
N-terminal sequencing 171
Analysis of secondary and tertiary structure 173
Endotoxin and other pyrogenic contaminants 173
Endotoxin, the molecule 174
Pyrogen detection 176
Dna 179
Microbial and viral contaminants 180
Viral assays 181
Miscellaneous contaminants 182
Validation studies 183
Further reading 185
Chapter 4 The cytokines -the interferon family 189
Cytokines 189
Cytokine receptors 194
Cytokines as biopharmaceuticals 195
The interferons 196
The biochemistry of interferon-a 197
Interferon-b 198
Interferon-g 198
Interferon signal transduction 198
The interferon receptors 199
The JAK-STAT pathway 199
The interferon JAK-STAT pathway 202
The biological effects of interferons 203
The eIF-2a protein kinase system 207
Interferon biotechnology 207
Production and medical uses of IFN-a 210
Medical uses of IFN-b 213
Medical applications of IFN-g 214
Interferon toxicity 216
Additional interferons 218
Conclusion 219
Further reading 219
Chapter 5 Cytokines: interleukins and tumour necrosis factor 223
Interleukin-2 (IL-2) 225
IL-2 production 228
IL-2 and cancer treatment 228
IL-2 and infectious diseases 230
Safety issues 231
Inhibition of IL-2 activity 231
Interleukin-1 (IL-1) 232
The biological activities of IL- 1 233
IL-1 biotechnology 234
Interleukin-3: biochemistry and biotechnology 235
Interleukin- 4 236
Interleukin- 6 238
Interleukin- 11 240
Interleukin- 5 241
Interleukin- 12 244
Tumour necrosis factors (TNFs) 246
TNF biochemistry 246
Biological activities of TNF-a 247
Immunity and inflammation 248
TNF receptors 249
TNF: therapeutic aspects 250
Further reading 252
Chapter 6 Haemopoietic growth factors 255
The interleukins as haemopoietic growth factors 257
Granulocyte colony stimulating factor (G-CSF) 258
Macrophage colony-stimulating factor (M-CSF) 259
Granulocyte-macrophage colony stimulating factor (GM-CSF) 259
Clinical application of CSFs 261
Leukaemia inhibitory factor (LIF) 263
Erythropoietin (EPO) 264
The EPO receptor and signal transduction 267
Regulation of EPO production 267
Therapeutic applications of EPO 268
Chronic disease and cancer chemotherapy 271
Additional non-renal applications 272
Tolerability 273
Thrombopoietin 273
Further reading 275
Chapter 7 Growth factors 277
Growth factors and wound healing 277
Insulin-like growth factors (IGFs) 279
IGF biochemistry 280
IGF receptors 280
IGF-binding proteins 282
Biological effects 282
IGF and fetal development 283
IGFs and growth 283
Renal and reproductive effects 284
Neuronal and other effects 285
Epidermal growth factor (EGF) 285
The EGF receptor 286
Platelet-derived growth factor (PDGF) 287
The PDGF receptor and signal transduction 288
PDGF and wound healing 289
Fibroblast growth factors (FGFs) 289
Transforming growth factors (TGFs) 290
TGF-a 290
TGF-b 292
Neurotrophic factors 293
The neurotrophins 294
Neurotrophin receptors 296
The neurotrophin low-affinity receptor 297
Ciliary neurotrophic factor and glial cell line-derived neurotrophic factor 297
Neurotrophic factors and neurodegenerative disease 298
Amyotrophic lateral sclerosis (ALS) and peripheral neuropathy 298
Neurotrophic factors and neurodegenerative diseases of the brain 298
Further reading 300
Chapter 8 Hormones of therapeutic interest 303
Insulin 303
Diabetes mellitus 304
The insulin molecule 304
The insulin receptor and signal transduction 307
Insulin production 307
Enzymatic conversion of porcine insulin 311
Production of human insulin by recombinant DNA technology 312
Formulation of insulin products 314
Engineered insulins 317
Additional means of insulin administration 320
Treating diabetics with insulin-producing cells 321
Glucagon 321
Human growth hormone (hGH) 324
Growth hormone releasing factor (GHRF) and inhibitory factor (GHRIF) 325
The GH receptor 325
Biological effects of GH 327
Therapeutic uses of GH 328
Recombinant hGH (rhGH) and pituitary dwarfism 328
Idiopathic short stature and Turner's syndrome 330
Metabolic effects of hGH 330
GH, lactation and ovulation 331
The gonadotrophins 331
Follicle stimulating hormone (FSH), luteinizing hormone (LH) and human chorionic gonadotrophin (hCG) 331
Pregnant mare serum gonadotrophin (PMSG) 335
The inhibins and activins 337
LHRH and regulation of gonadotrophin production 338
Medical and veterinary applications of gonadotrophins 339
Sources and medical uses of FSH, LHand hCG 340
Recombinant gonadotrophins 342
Veterinary uses of gonadotrophins 344
Gonadotrophin releasing hormone (GnRH) 345
Additional recombinant hormones now approved 345
Conclusions 348
Further reading 348
Chapter 9 Blood products and therapeutic enzymes 351
Disease transmission 351
Whole blood 353
Platelets and red blood cells 353
Blood substitutes 353
Dextrans 354
Albumin 355
Gelatin 357
Oxygen-carrying blood substitutes 357
Haemostasis 358
The coagulation pathway 358
Terminal steps of coagulation pathway 361
Clotting disorders 365
Factor VIII and haemophilia 366
Production of Factor VIII 368
Factors IX, VII a and XIII 371
Anticoagulants 372
Heparin 372
Vitamin Kantimetabolites 375
Hirudin 375
Antithrombin 379
Thrombolytic agents 380
Tissue plasminogen activator (tPA) 381
First-generation tPA 383
Engineered tPA 383
Streptokinase 385
Urokinase 386
Staphylokinase 386
a1 -Antitrypsin 388
Enzymes of therapeutic value 389
Asparaginase 390
DNase 392
Glucocerebrosidase 393
a-Galactosidase and urate oxidase 395
Superoxide dismutase 397
Debriding agents 397
Digestive aids 398
Lactase 400
Further reading 400
Chapter 10 Antibodies, vaccines and adjuvants 403
Polyclonal antibody preparations 403
Anti-D immunoglobulin 406
Normal immunoglobulins 407
Hepatitis Band tetanus immunoglobulin 407
Snake and spider antivenins 408
Monoclonal antibodies 409
Production of monoclonals via hybridoma technology 411
Antibody screening: phage display technology 412
Therapeutic application of monoclonal antibodies 414
Tumour immunology 415
Antibody-based strategies for tumour detection/destruction 417
Drug-based tumour immunotherapy 424
First-generation anti-tumour antibodies: clinical disappointment 426
Tumour-associated antigens 426
Antigenicity of murine monoclonals 428
Chimaeric and humanized antibodies 429
Antibody fragments 432
Additional therapeutic applications of monoclonal antibodies 433
Cardiovascular and related disease 433
Infectious diseases 433
Autoimmune disease 434
Transplantation 434
Vaccine technology 435
Traditional vaccine preparations 436
Attenuated, dead or inactivated bacteria 438
Attenuated and inactivated viral vaccines 439
Toxoids, antigen-based and other vaccine preparations 440
The impact of genetic engineering on vaccine technology 441
Peptide vaccines 444
Vaccine vectors 445
Development of an AIDS vaccine 447
Difficulties associated with vaccine development 450
AIDS vaccines in clinical trials 450
Cancer vaccines 452
Recombinant veterinary vaccines 452
Adjuvant technology 453
Adjuvant mode of action 455
Mineral-based adjuvants 455
Oil-based emulsion adjuvants 455
Bacteria/bacterial products as adjuvants 457
Additional adjuvants 458
Further reading 460
Chapter 11 Nucleic acid therapeutics 463
Gene therapy 463
Basic approach to gene therapy 464
Some additional questions 467
Vectors used in gene therapy 468
Retroviral vectors 468
Additional viral-based vectors 472
Manufacture of viral vectors 474
Non-viral vectors 476
Manufacture of plasmid DNA 480
Gene therapy and genetic disease 482
Gene therapy and cancer 485
Gene therapy and AIDS 486
Gene-based vaccines 488
Gene therapy: some additional considerations 488
Anti-sense technology 488
Anti-sense oligonucleotides 490
Uses, advantages and disadvantages of 'oligos' 491
Delivery and cellular uptake of oligonucleotides 493
Manufacture of oligonucleotides 493
Vitravene, an approved antisense agent 494
Antigene sequences and ribozymes 494
Conclusion 495
Further reading 496
Appendix 1 Biopharmaceuticals thus far approved in the USA or European Union 499
Appendix 2 Some Internet addresses relevant to the biopharmaceutical sector 509
Appendix 3 Two selected monographs reproduced from the European Pharmacopoeia with permission from the European Commission: I. Products of recombinant DNA technology 515
II. Interferon a-2 concentrated solution 520
Appendix 4 Manufacture of biological medicinal products for human use. (Annex 2 from The Rules Governing Medicinal Products in the European Community, Vol. 4, Good Manufacturing Practice for Medicinal Products) 527
Index 533
