BỘ Y TẾ TRƯỜNG ĐẠI HỌC DƯỢC HÀ NỘI NGUYỄN THỊ THÚY MÃ SINH VIÊN: 1501484 THE CURRENT STATUS OF mRNA AND OTHER VACCINES MANUFACTURING TECHNOLOGIES IN VIETNAM KHÓA LUẬN TỐT NGHIỆP DƯỢC SĨ Người hướng dẫn: 1. Đàm Thanh Xuân 2. Nguyễn Khắc Tiệp Nơi thực hiện: Bộ môn Công nghiệp dược HÀ NỘI – 2022 ACKNOWLEDGEMENT Foremost, I would like to express my sincere gratitude and give my warmest thanks to my supervisors and Associate Professor Dr. Dam Thanh Xuan and Dr.
Nguyen Khac Tiep who made this work possible. Their guidance, support and advice carried me through all the stages of writing my project. I could not have undertaken this journey without their help and patience. I would also like to thank school committee and professors for teaching, helping and especially inspiring me to see the values that pharmacists can create and contribute to the society.
Thanks to all professors and lecturers with their dedication and support, my 5-year program at Hanoi University of Pharmacy cannot be more amazing and precious. Finally, I would like to give special thanks to my family and friends as a whole for their continuous support and understanding. Due to the limited time and shortage of my knowledge, the thesis may have shortages. I sincerely hope to have your feedback on my thesis to make it better.
Best regards, Hanoi, June 2022 Student Nguyen Thi Thuy TABLE OF CONTENTS ACKNOWLEDGEMENT TABLE OF CONTENTS LIST OF ABBREVIATIONS LIST OF TABLES LIST OF FIGURES ABSTRACT. VACCINE DEFINITION AND CLASSIFICATION. VACCINE MANUFACTURING TECHNOLOGY OVERVIEW IN VIETNAM. Live attenuated vaccine (LAV) technology.
Measles vaccine vaccine trade name: MVVAC). Measles-rubella vaccine (vaccine trade name: MRVAC). Inactivated vaccine technology. Japanese encephalitis vaccine (vaccine trade name: Jevax®).
Toxoid vaccine technology. Subunit vaccine technology. mRNA TECHNOLOGY AND CURRENT STATUS OF mRNA TECHNOLOGY IN VACCINE MANUFACTURING IN VIETNAM. History of mRNA vaccine.
Applications of mRNA vaccine. Advantages and disadvantages of mRNA vaccine. mRNA vaccine classification. Mechanism of immune response induced by mRNA vaccines.
mRNA vaccine technology .The manufacturing process. Conventional and self-amplifying mRNA design. Lipid nanoparticles (LNP) technology. COVID-19 and mRNA COVID-19 vaccine.
mRNA technology in COVID-19 vaccine manufacturing in Vietnam. STARRTM mRNA technology. LUNAR® delivery system. Advantages and Disadvantages of ARCT-154.
mRNA VACCINE IN THE FUTURE .38 CONCLUSION AND RECOMMENDATION. 40 REFERENCES LIST OF ABBREVIATIONS Abbreviation Meaning APC Antigen-presenting cell ATX Arcturus BCG Bacille Calmette-Guérin CCID50 Cell culture infectious dose 50 CD4, 8 Cluster of differentiation 4, 8 CDC Centers for Disease Control and Prevention CDS Coding sequence CHO Chinese hamster ovary CIGB Center for Genetic Engineering and Biotechnology COVID-19 Coronavirus disease of 2019 CQA Critical quality attribute CTL Cytoxic T lymphocyte DAVAC Vaccine Company Limited of Dalat Pasteur DC Dendritic cell DMEM Dulbecco’s Modified Eagle’s DNA Deoxyribonucleic Acid DOPE Dioleoylphosphatidylethanolamine DOTAP 1,2-dioleoyloxy-3-trimethylammoniumpropane DPT Diptheria, tetanus toxoids and pertussis dsRNA Double-stranded RNA EPO Erythropoetin E.LPS Endotoxin Units lipopolysaccharide FDA Food and Drug Administration FFU Fluorecent Focus-Forming Unit FPLC Fast protein liquid chromatography HbsAg Hepatitis B surface antigen HIV Human immunodeficiency virus HPLC High-performance liquid chromatography HPV Human papillomavirus GAVI Global Alliance for Vaccines and Immunization GFP Green fluorescent protein GMP Good Manufacturing Practice IFN Interferon IFIT1 IFN-induced proteins with tetratricopeptide repeats IPV Inactivated polio vaccine IVAC Institute of Vaccines and Medical Biologicals IVT In vitro transcribed LAV Live attenuated vaccine LNP Lipid nanoparticle MDA5 Melanoma differentiation-associated gene 5 MERS-CoV Middle East Respiratory Syndrome MMR Measles, mumps and rubella mRNA Messenger RNA MSV Master seed virus MS Master seed NOD2 Nucleotide Binding Oligomerization Domain Containing 2 NRM Non-replicating Mrna ORF Open-reading frame pDNA Plasmid DNA PEG Polyethylene glycol PEI Polyethylenimine PFU Plaque-forming unit pKa The acid dissociation constant PKR Protein kinase R Pmkc Primary monkey kidney cells POLYVAC Center for Research and Production of Vaccines and Biologicals RIG-I Retinoic acid-inducible gene I RNA Ribonucleic acid RT-PCR Real time - polymerase chain reaction SAM Self-amplifying Mrna SARS-CoV-2 Severe Acute Respiratory Syndrome 2 SARS-CoV Severe Acute Respiratory Syndrome siRNA Small interfering RNA SPF Specific Pathogen Free ssRNA Single-stranded RNA TAA Tumor-associated antigen TFH T follicular helper TLR Toll-like receptor TNF Tumor Necrosis Facto TSA Tumor-specific antigen USA United States of America UTR Untranslated region UV Ultra-Violet VABIOTECH Vaccine and Biological Production Company No. 1 WSV Working seed virus WS Working seed LIST OF TABLES Table 2. Six Vietnam Vaccine Manufacturers…………………………………….
Six Vietnam Vaccine Manufacturers and their Products………………. Primary outcome measures…………………………………………….36 LIST OF FIGURES Figure 3. Development timeline of mRNA vaccine [38], [44], [45]………………. Two categories of mRNA constructs from designing stage to antigen expressing stage [54]…………………………………………………………….
How mRNA vaccines elicit immunity [59]……………………………. Five critical quality attributes (CQAs) dictating the performance of mRNA construct to express the gene of interest efficiently [54]……………………………. In vitro transcription reaction component [80]…………………………. Major LNP and polymer delivery methods for mRNA vaccines……….
COVID-19 mRNA vaccine manufacturing process [133]………………. Arcturus’s COVID-19 vaccine technology…………………………….31 ABSTRACT The impact of vaccines in reducing the health burden of infectious diseases has been one of the greatest achievements in public health [1]. Smallpox, a disease which caused more than 300 million deaths since 19th century [2], has been eradicated in the world [1]. Although smallpox is the only human disease which has been eradicated, with the advances in technology, there are more diseases which are believed and targeted for global eradication in the near future.
Advances in vaccine manufacturing technology plays an important role in allowing more countries to be able to produce vaccines as well as helping scientists to invent more vaccines, thereby leading to millions of lives could be saved from infectious diseases. Up to now, mRNA vaccine is the latest type of vaccine which is invented and approved. There are only three approved COVID-19 mRNA vaccines; however, there are 50 mRNA vaccines which are in clinical trials [3]. That a lot of mRNA vaccines are being researched, not only for COVID-19 but also for other diseases shows the huge potential of this new vaccine type.
As a Global Alliance for Vaccines and Immunization (GAVI) – eligible country, Vietnam has been able to produce the majority of vaccines used in the National Immunization Program [4]. Currently, Vietnam is also executing clinical trials in COVID-19 mRNA vaccines [5]. Therefore, Vietnam is a country that is proactively producing a lot of vaccine types requiring different manufacturing technologies. However, there is no any official document about the current status of vaccine manufacturing technologies being used and mRNA vaccine technology being researched in Vietnam.
Therefore, this thesis is written with these three main purposes: 1. Summarize the current status of vaccine manufacturing technologies used in all Vietnamese vaccine manufacturers. Provide the overview of the mRNA vaccine technology. Provide the basic information about the mRNA vaccine being researched in Vietnam.
VACCINE DEFINITION AND CLASSIFICATION 1. Definition According to Vietnam Pharmacopeia 5th edition, human vaccine is defined as a preparation containing antigens which can stimulate adaptive immune response to prevent diseases. Vaccines are produced from bacteria, Rickettsia or viruses. Human vaccine includes: live attenuated, inactivated microbes and antigenic components of the pathogens.
However, with the fast development of the vaccine manufacturing field, especially with the birth of mRNA vaccines and other modern vaccines, this definition appears to be not appropriate and updated. According to the USA CDC, vaccine is a preparation that is used to stimulate the body’s immune response against diseases. Vaccines are usually administered through needle injections, but some can be administered by mouth or sprayed into the nose [6]. Classification Vaccines can be classified based on different criteria as follows.
Based on effects, there are single vaccines and combination vaccines. A single vaccine is used for only one disease (eg: BCG vaccine, influenza vaccine,…), whereas, a combination vaccine is two or more different vaccines that have been combined into a single shot and the combined effect is at least the same or better than the single vaccine (eg: DPT vaccine, MMR vaccine,…) [7]. Based on pathogens, there are virus vaccines (eg: polio vaccine, COVID-19 vaccine,…) and bacterial vaccines (BCG vaccine, cholera vaccine,…). Based on administration routes, there are oral vaccines (eg: polio vaccine), injectable vaccines (MMR vaccine) and others such as inhalable,… Based on generation, there are first generation vaccine (live attenuated vaccine or LAV, inactivated vaccine and toxoid vaccine), second generation vaccine (protein subunit vaccine and recombinant vector viral vaccine) and third generation vaccine (combination of recombinant vaccine and new excipients in order to stimulate desired immune response).
Based on history, there are traditional/conventional vaccines (LAV, inactivated vaccine, toxoid vaccine, subunit vaccine) and modern vaccines (LAV produced by gene technique, recombinant antigen, DNA vaccine,…) 2 CHAPTER 2. VACCINE MANUFACTURING TECHNOLOGY OVERVIEW IN VIETNAM To date, in Vietnam, there are four vaccine manufacturers which are VABIOTECH, POLYVAC, IVAC and DAVAC. These four manufacturers have manufactured the majority of vaccines used in Expanded Program on Immunization (EPI) in Vietnam. Besides, there are two new companies which are doing research on COVID-19 vaccines as their very first vaccine which sets a milestone of their entry into vaccine manufacturing field.
Below are these manufacturers with their products. Six Vietnam Vaccine Manufacturers Name Full names in English Full names in Vietnamese abbreviations VABIOTECH Vaccine and Biological Công ty TNHH MTV Vacxin và Production Company No. 1 Sinh phẩm số 1 – Bộ Y Tế (Hà Nội) POLYVAC Center for Research and Trung tâm nghiên cứu và sản xuất Production of Vaccines and vắc xin và sinh phẩm y tế (Hà Nội) Biologicals IVAC Institute of Vaccines and Medical Viện Vắc Xin Và Sinh Phẩm Y Tế Biologicals (Nha Trang) DAVAC Vaccine Company Limited Công ty TNHH Một Thành Viên of Dalat Pasteur Vắc xin Pasteur Đà Lạt (Đà lạt) VinBioCare VinBioCare Biotechnology Joint Công ty cổ phần Công nghệ sinh học Stock Company VinBioCare (Hà Nội) 3 Nanogen Nanogen Pharmaceutical Công ty cổ phần Công nghệ sinh học Biotechnology Joint Stock dược Nanogen Company Table 2. Six Vietnam Vaccine Manufacturers and their Products Manufacturers Products Status VABIOTECH Oral cholera, japanese Approved encephalitis, hepatitis B, hepatitis A vaccines POLYVAC Rota virus, measles, polio, Approved measles-rubella vaccines IVAC BCG vaccine, pertussis antigen in Approved DPT vaccine; diphtheria-tetanus vaccine DAVAC Typhoid vaccine Approved VinBioCare Covid-19 vaccine Clinical trial (not approved yet) Nanogen Covid-19 vaccine Clinical trial (not approved yet) According to the table above, each vaccine manufacturer is producing different vaccines.
However, all of these vaccines fall into 4 categories: live-attenuated, inactivated, toxoid and subunit vaccines. This chapter will discuss the technologies used in each vaccine type and the detailed information of each product provided by the manufacturers. Live attenuated vaccine (LAV) technology 2. Overview Live, attenuated vaccines contain a version of the living pathogenic microbe that has been attenuated or weakened in the lab so that it has lost its significant pathogenicity [8].
Manufacturing principle: bacteria or viruses are cultured in special conditions in order to weaken or remove their pathogenicity, then live attenuated cells are collected. There are two kinds of live attenuated vaccines. The first kind of LAV is produced by isolating pathogenic microbes from animals whose diseases are similar to humans’ diseases. Therefore, this type of vaccine can stimulate protective immune responses without causing diseases in humans.
The second kind of LAV is produced by isolating pathogenic microbe from patient, then culturing them in vitro in cells and finally, weakening them by different methods such as heat method for heat-sensitive microbes (oral Sabin polio vaccines,…) and biological method of serial passages through a foreign host to mutate genes (measles vaccines,…) [8]. According to vaccine classification based on history, this is the traditional vaccine. Therefore, one disadvantage is the fact that this vaccine requires a culture process. Moreover, microbes in LAV are live which means that there is potential for the vaccine virus to revert to a form capable of causing disease to humans, especially immunocompromised people.
This vaccine also requires to be kept cool.