1. FAST TRACKING THE DESIGN AND DEVELOPMENT FOR VACCINES FOR “NEGLECTED DISEASES”
DR GAURAV GUPTA ,
Head Viral and genetic engineered vaccines, Vaccine technology centre, Zydus Cadila

2. CONTENTS Introduction
Challenges in developing new generation of neglected disease vaccines
Novel approaches for the design of NTD vaccines- 2 case studies
Need for developing new technologies to address neglected disease vaccines

3. INTRODUCTION

4. Definition
Neglected tropical diseases (NTDs) are a diverse group of communicable diseases that prevail in tropical and subtropical conditions in 149 countries and affect more than one billion people, costing developing economies billions of dollars every year.
They mainly affect populations living in poverty, without adequate sanitation and in close contact with infectious vectors and domestic animals and livestock.

5. List of Neglected Tropical Diseases
HELMINTH INFECTIONS:
Ascariasis, trichuriasis, hookworm infection, strongyloidiasis, toxocariasis and larva migrans, lymphatic filariasis, onchocerciasis, loiasis, dracunculiasis, schistosomiasis, food-borne trematodiases, taeniasis, cysticercosis, echinococcosis
BACTERIAL INFECTIONS:
Bartonellosis, bovine tuberculosis, buruli ulcer, leprosy,leptospirosis, relapsing fever, rheumatic fever, trachoma, treponematoses

6. List of Neglected Tropical Diseases
FUNGAL INFECTIONS:
mycetoma, paracoccidiomycosis
PROTOZOAN INFECTIONS:
leishmaniasis, Chagas disease, human African trypanosomiasis, amoebiasis, giardiasis, balantidiasis
VIRAL INFECTIONS:
dengue fever, yellow fever, Japanese encephalitis, rabies, haemorrhagic fevers
ECTOPARASITIC INFECTIONS:
scabies, myiasis, tungiasis

7. WHO LIST OF NEGLECTED TROPICAL DISEASES (NTDS)- 17 DISEASES

8. Global Overlap of Six of the Common NTDs
6 NTDs: guinea worm disease, lymphatic filariasis, onchocerciasis, schistosomiasis, soil-transmitted helminths, trachoma

9. Neglected by whom Producers of drugs and diagnostic tests
National policy makers
Funders (R&D and control programs)
Academic centers
Media

10. Landmarks in overcoming NTDs

11. Growing R&D Pipelines by Industry and partners for NTDs
Source- IFPMA

12. MAJOR RESEARCH INSTITUTES INVOLVED in NTDs VACCINES
Sabin Vaccine Institute, US
Infectious disease research Institute, US (IDRI)
Industrial federation of pharmaceutical manufacturers and associations (IFPMA)

13. MAJOR CHALLENGES FOR VACCINE DEVELOPMENT
Limited funding
Market potential and payback scenario
Big multinational companies are not attracted.
Requires extensive clinical trial without surrogate of protection.
Lack of easy to use, low cost diagnostic

14. TECHNICAL CHALLENGES TO DEVELOP VACCINES
ANTIGEN DISCOVERY
Complicated genetic structures of NTD pathogens and high genetic diversity
Absence of genome databases or bioinformatic algorithms for selecting candidate antigens of promise.
Parasites have mutistage life cycle

15. TECHNICAL CHALLENGES TO DEVELOP VACCINES
PROCESS DEVELOPMENT
Necessity to scale up production of NTDs vaccine at adequate yields and at low cost.
Failure of many bacterial expression systems to produce properly folded recombinant proteins (helminth antigens)
Cysticercosis and Echinococcosis in E coli were very successful for antigen expression

16. TECHNICAL CHALLENGES TO DEVELOP VACCINES
PRECLINICAL DEVELOPMENT
Difficulty in maintaining cycle stages of NTD pathogens in vitro.
Paucity of laboratory animal models permissive to the NTD pathogens or that can accurately reproduce human disease or protective immunity.
CLINICAL DEVELOPMENT
Clinical trials in resource-poor settings.
Highly modulated immune response from infection with many NTDs, especially helminth NTDS, present concerns like allegry

17. NOVEL APPROACHES FOR VACCINE DESIGN FOR NTDs

18. Vaccine design for NTDs
Selection of Vaccine technologies
Live
Inactivated
Recombinant protein (Mostly)
Polysaccharide protein conjugate
Viral vectored ( Dengue)
DNA vaccines
Peptide based vaccines

19. VACCINE DEVELOPMENT PLAN
Selection of multistage antigens (parasitic vaccines) using reverse vaccinology approaches
Selection of appropriate adjuvants
Preclinical studies with challenge animal models for efficacy or in vitro neutralization studies
Identification of immune correlates ( antibodies or cellular immune response or both )
Conducting clinical trials for safety and efficacy (I,II and III)

20. APPROACHES FOR NTDs
Multiple antigens vaccine targetting different stages of life cycle
Parasites can be blocked at infection, growth and transmission
Combining NTD vaccine development with other priority vaccines in pipeline to de-risk investment.

21. APPROACHES FOR NTDs
Involving funding agencies for research and development.
Using low cost in house vaccine platform technologies
Collaborations- Public private or private- private

22. Leishmania Vaccine Development- A case study 1

23. Life cycle of Leishmania
confidential

24. PROTECTIVE IMMUNE RESPONSES IN THE HOST
Various Vaccine candidates have been studied in animal models till date.
In Mice, Protective immunity to Leishmania infection requires the development of interleukin-12-dependent, parasite-specific Th1 responses, characterized by interferon-ᵧ and tumor necrosis factor production by CD4+ T cells.
These cytokines are required for the generation of reactive oxygen and nitrogen species by infected macrophages that enables killing of intracellular parasites.
confidential

25. PROTECTIVE IMMUNE RESPONSES IN THE HOST
Recent advances have also been made in understanding immunoregulatory mechanisms and observed suppression of parasite-specific CD4+ T-cell responses in human VL patients.
Also correlated with interleukin-10 produced by CD4+ T cells is a potent, autocrine inhibitor of interferon-ᵧ production and promotes parasite persistence in spleen tissue from VL patients.
Thus, interleukin-10 has been identified as a potential therapeutic target.
confidential

26. THE IDEAL LEISHMANIA VACCINE CHARACTERSTICS
Safe
Induces effective T cell response against appropriate antigens
Induces long-term immunity
Prophylactic and therapeutic activity
Effective against more than one form of leishmaniasis
Cost-effective
Reproducible, transferable manufacturing process
confidential

27. LEISHMANIA VACCINE DEVELOPMENT APPROACH
Selection of multistage antigens with established proof of concept studies in animal models and /or Clinical trials
in preclinical challenge animal models (Mice and Hamsters)
Optimizing potent formulations (Antigens +adjuvants )
Using best vaccine candidates for human clinical trials for safety & efficacy
confidential

28. CURRENT STATUS
European Research Center of Zydus Cadila has won highly competitive Frame work program 7 projects to develop a new vaccine against Leishmaniasis
Collaboration centers include Universidad Autónoma de Madrid, Spain; Instituto de Salud Carlos III, Spain; Instituto de Biologia Molecular e Cellular, Portugal; Istituto Superiore di Sanità, Italy; Swiss Tropical Public Health Institute, Switzerland; Meditox, Czech Republic; AMVAC Ag, Switzerland; Sabin Institute, Texas, NIH in Bethesda & IDRI in Seattle, USA
Overall Budget sponsored by European Union 6.0 million Euros
This Project foresee the phase I/II clinical trial in humans
confidential

29. Evaluation of vaccine candidates funded by BIRAC, New Delhi and Zydus
ANIMAL STUDIES
To evaluate proposed vaccine formulations in rodent models and Rhesus monkey experiments
HUMAN STUDIES
To evaluate in vitro cellular immune response in Leishmaniasis with and without our proposed vaccine formulations from active VL patients, cured individuals and healthy endemic control subjects from their PBMCs (BUS, Patna and AIMS, NEW DELHI)
Final potential candidate Vaccine formulations will be further evaluated in clinical trials for safety and efficacy by Zydus.
confidential

30. Collaboration with IDRI, US
confidential

31. confidential

32. FUTURE PLANS ON VL VACCINES
To generate clinical data based on EU project and to plan Indian Clinical trials accordingly for safety & efficacy
To commercialize a affordable, safe and effective vaccine for India and Global use as soon as possible
confidential

33. Chikungunya Vaccine Development- Case Study 2

34. Chikungunya Vaccine development

35. Takeda’s Candidate Chikungunya Vaccine
Inactivate subgenomic promoter :
Insert the internal ribosome entry site (IRES) of encephalomylocarditis virus
13 synonymous mutations
Attenuating phenotypes:
Reduces expression of capsid and envelope proteins
Prevents viral expression in mosquito cells: no recognition of IRES
Reference: IBC_Agent_3_CHIK_2015_update.pdf
Plante et al., PLoS Pathogens

36. Takeda’s Chikungunya Vaccine Candidate: Attenuation In Vitro
Analyze levels of Chikungunya vaccine RNA replication and structural protein expression in vitro.
Determine sites and patterns of CHIK vaccine replication and spread in vaccinated mice
Reference: v3.0 Molecular Cloning of CHIK Vaccine w Signature.pdf
Plante et al., PLoS Pathogens

37. Chikungunya vaccine development approach
Process and analytical development
Preclinical studies
Clinical studies
Final commcerialization

38. MEASLES VECTORED VACCINE CANDIDATES- REVERSE GENETICS APPRAOCH

39. Measles Reverse Genetics technology

40. CLONING AND RESCUE OF ANTIGENS IN MEASLES VIRUS VECTOR
p(+)MV
293 cell line with
T7 RNA pol
N and P T7
pEMCLa
Cotransfection
Syncytia formation
Virus rescue
EMC
IRES L

41. NEED FOR NEW TECHNOLOGIES
Rapid and cost effective process development
Quick and cheap diagnostics
Genomics based reverse vaccinology tools for vaccine candidate selection
Transgenic animal models for proof of concept
Compact, multiproduct GMP facilities for reducing cost of production
Rapid immunoassays and potency assays

42. Summary
Collaboration is the key to achieve NTD vaccines in short time with affordable price.
Multistage and multiantigens for parasitic vaccines are key immune response targets.
Attracting funding agencies to reduce risk on product development should be way forward.
Developing preclinical animal models can accelerate the vaccine development.
Genomics and proteomics tools should be drivers for identification and selection of vaccine candidates.

43. THANK YOU gauravgupta@zyduscadila.com drgaurav123@gmail.com