mRNA Therapeutics: Fast-to-Market Strategies

327 Pages·2022·40.678 MB·English

by  Sarfaraz K. Niazi

mRNA Therapeutics
This book is the most comprehensive and complete treatise on nucleic acid thera-
peutic products, including mRNA vaccines, their manufacturing, formulations, and
testing for safety and efficacy. Details include cGMP-compliant manufacturing and
regulatory filing steps. A new concept of “biosimilar” mRNA vaccine is presented
to secure fast approval of copies of mRNA vaccines. Projections of financial plans
to establish RNA manufacturing facilities are provided, along with details of supply
chain management. Finally, the future of nucleic acid products in gene therapy and
other newer applications is presented, along with a perspective that all new vaccines
will be the nucleic acid type that will further provide first-time prevention of autoim-
mune disorders. It is projected that both big pharma and start-ups will enter this field,
and we can expect significant additions to our drug armamentarium soon.

mRNA Therapeutics
Fast-to-Market Strategies
Sarfaraz K. Niazi
First edition published 2022
by CRC Press
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and by CRC Press
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CRC Press is an imprint of Taylor & Francis Group, LLC
© 2023 Sarfaraz K. Niazi
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ISBN: 9781032163444 (hbk)
ISBN: 9781032163482 (pbk)
ISBN: 9781003248156 (ebk)
DOI: 10.1201/9781003248156
Typeset in Times
by Deanta Global Publishing Services, Chennai, India
To: Johannes Friedrich Miescher (13 August 1844–26 August
1895), a Swiss physician and biologist who first isolated nucleic
acid in 1869, paving the way for the identification of DNA as
the carrier of inheritance as he suggested a possible role of
the nucleic acids in heredity and evolution. The Nobel Prize
was not established then, and it is not given posthumously.

Contents
Preface and Perspective ...........................................................................................xv
Acknowledgments ...................................................................................................xix
Author .....................................................................................................................xxi
Background ..............................................................................................................1
B.1 The RNA Lead ..........................................................................4
B.2 Development perspective .........................................................14
B.3 Intellectual Property Perspective ............................................15
B.4 Publications .............................................................................17
Chapter 1 The Genome Machine ........................................................................19
1.1 Background ..............................................................................19
1.2 Molecular Clock ......................................................................21
1.3 Evolution ..................................................................................22
1.4 Life ..........................................................................................24
1.4.1 Aging ..........................................................................28
1.4.2 Humans ......................................................................29
1.4.3 Biological Cell ............................................................29
1.5 Proteins ....................................................................................34
1.6 Conclusion ...............................................................................39
Chapter 2 Understanding Nucleic Acids .............................................................41
2.1 Background ..............................................................................41
2.2 The DNA .................................................................................42
2.3 The RNA .................................................................................47
2.3.1 Replication ..................................................................52
2.3.2 Transcription ..............................................................54
2.3.3 Codons ........................................................................58
2.3.4 Translation Process ....................................................59
2.4 Leventhal Paradox ...................................................................65
2.5 Conclusion ...............................................................................66
Chapter 3 RNA Therapeutics ..............................................................................67
3.1 Background ..............................................................................67
3.2 DNA Therapeutics ...................................................................68
3.2.1 DNA Plasmids ............................................................68
3.2.2 Viral Vectors ..............................................................68
vii
viii Contents
3.3 Types of RNAs ........................................................................70
3.4 Approved Therapies ................................................................70
3.5 RNA Therapies ........................................................................72
3.6 Noncoding RNA ......................................................................76
3.6.1 Small interfering RNA ...............................................79
3.6.2 Micro RNA .................................................................81
3.6.3 Long Non-coding RNA ..............................................82
3.6.4 Antisense RNA ...........................................................83
3.6.5 Interfering RNA .........................................................83
3.6.6 Aptamers ....................................................................83
3.7 Coding RNA ............................................................................86
3.7.1 mRNA as Therapeutics ..............................................86
3.7.2 mRNA as Replacement Therapy ................................87
3.7.3 mRNA Vaccines .........................................................89
3.7.4 mRNA-Enhanced Cell Therapies ..............................91
3.8 Delivery of RNA Therapeutics ................................................94
3.8.1 Lipid Nanoparticles ....................................................94
3.8.2 Liposomes ..................................................................96
3.8.2.1 Leukosomes ................................................97
3.8.3 Nanotechnology..........................................................98
3.8.3.1 Therapeutic Modules ..................................98
3.8.3.2 Construction of Multifunctional RNA
Nanoparticles ............................................101
3.8.4 Polymer Nanomaterials ............................................102
3.8.5 Silica Nanoparticles..................................................103
3.8.6 Carbon and Gold Nanomaterials ..............................103
3.8.7 N-Acetylgalactosamine (GalNAc) ............................104
3.8.7.1 Testing .......................................................105
3.9 Conclusion .............................................................................106
Chapter 4 Nucleoside Vaccines .........................................................................107
4.1 Background ............................................................................107
4.2 History ...................................................................................107
4.3 Immunogenicity .....................................................................110
4.3.1 Antigens....................................................................110
4.3.2 Dendritic Cells .........................................................112
4.3.3 RNA Virus ...............................................................112
4.4 Messenger RNA ....................................................................113
4.4.1 Types ........................................................................114
4.4.2 Chemical Modifications ...........................................116
4.4.2.1 Sugar Modifications ..................................116
4.4.2.2 Backbone Modifications ...........................117
4.4.2.3 Base Modifications: ..................................117
Contents ix
4.4.3 MHC .........................................................................119
4.4.4 Stability ....................................................................119
4.4.5 Production ................................................................123
4.4.6 Delivery ....................................................................123
4.4.6.1 Lipid Nanoparticles ..................................127
4.5 Development Strategies .........................................................131
4.5.1 Vaccine Design .........................................................131
4.5.2 Process Development ...............................................133
4.5.3 Construction of Plasmids for RNA Transcription ....136
4.5.4 Plasmid Purification and Linearization ....................137
4.5.5 In Vitro Transcription of RNA .................................137
4.5.6 RNA Capping ...........................................................137
4.5.7 RNA Quality Control ...............................................138
4.5.8 Immunization and Allergic Sensitization of Mice ...138
4.5.9 Measurement of Antibody Subclasses .....................138
4.5.10 Basophil Release Assay ............................................139
4.5.11 Culture of RBL Cells ...............................................139
4.6 Nucleoside Vaccines Perspective ...........................................141
4.6.1 SARS-CoV-2 Vaccine ..............................................141
4.6.2 Antiallergy mRNA Vaccine .....................................149
4.6.3 Anti-infective mRNA Vaccines ...............................149
4.6.4 Rabies .......................................................................150
4.6.5 Influenza ...................................................................150
4.6.6 Respiratory Syncytial Virus .....................................151
4.6.7 Human Metapneumovirus and Type 3
Parainfluenza Virus ..................................................151
4.6.8 Human Cytomegalovirus .........................................151
4.6.9 Zika Virus ................................................................152
4.6.10 Epstein–Barr Virus ..................................................152
4.6.11 HIV ...........................................................................152
4.6.12 Streptococcus ...........................................................153
4.6.13 Ebola Virus ...............................................................153
4.7 Conclusion .............................................................................153
Chapter 5 cGMP mRNA Vaccine Manufacturing ............................................155
5.1 Background ............................................................................155
5.2 cGMP Process .......................................................................157
5.3 Manufacturing Process ..........................................................158
5.3.1 Protocol 1 Steps ........................................................158
5.3.1.1 IVT Steps ..................................................159
5.3.1.2 Final Quality Testing Steps: .....................159
5.3.2 Protocol 2 Steps ........................................................159
5.3.3 Facility ......................................................................161