Biopharmaceuticals

Gary Walsh is a senior lecturer in industrial biochemistry at the University of Limerick, Ireland. He combines industrial experience within the pharmaceutical industry with an extensive teaching and academic research background in pharmaceutical biotechnology. He has published half a dozen highly acclaimed books on various aspects of protein biotechnology. He is on the editorial board of several biopharmaceutical journals and is closely affiliated with the European Association of Pharma Biotechnology, serving as its scientific secretary.

"…contains just about everything that anyone would want to know about the subject…It's all here in this easy-to-read textbook.” (Biochemistry and Molecular Education, March/April 2004) "...well written… (and) copiously illustrated..." (Chemistry & Industry, 17th January 2005)“This book should be recommended reading for all under-graduate course in pharmacy and the pharmaceutical sciences …”. (Cell Biochemistry & Function, March-April 2005)

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