This is a final presentation made on a hypothetical vaccine namely Cholonex. The process of development and discovery covers all the aspects of drug making and might help students in understanding the complete proprietary drug or biologics making process.

1. BIOT 5120
12/10/2019
GROUP 8
Anjali Chandani
Shreya Mool
Namuun Erdenepurev
Marco Menara
Tom Pu
Xiaoheng Zhang

2. INTRODUCTION
Cholera
Acute intestinal infection
-- causes diarrhea and
dehydration, even death
• 74,000 and 595,000
(1990-2016)
• 1.3 and 4.0 million
cases
• 95,000 deaths yearly

3. INTRODUCTION
Vibrio Cholerae
• Toxin : Cholera Toxin (CTX)
• Toxin Coregulated pilus (TCP)
our target
• Other Proteins:
o Hemagglutinin
o Accessory colonization factor

4. MoA
Intestinal
walls
TCP
Toxin Coregulated
pilus
TARGET

5. DRUG LANDSCAPE

6. VAXCHORA:
THE ONLY APPROVED
VACCINE FOR CHOLERA IN
US

7. Vaxchora First FDA approved cholera vaccine
in the USA
Indications and usage •Active immunization against bacterium V.cholerae
sergroup O1
•18-64 years of age
Mechanism of action •Contains a live attenuated cholera bacterium that
replicates in the gastrointestinal tract of the receipt
Dosage and administration •A single dose must be administered a minimum of
10 days before potential exposure to cholera or
travel to cholera-endemic areas
Cost $270
DRUG LANDSCAPE

8. DRUG LANDSCAPE
Obstacles the current drugs are facing:
1.Perceived lack of profitable markets by vaccine
producers.
2.No introduction of available vaccines due to
uncertainty.
3.Available vaccines not made for use in developing
countries.
4.Most of current drugs target mainly travelers from
developed country.

9. COMMERCIAL NAME CHOLONEX
Components TcpA subunit from classical and El Tor
Cholera biotypes and recombinant cholera
toxoid
Exclusion criteria Children < 2 years
Presentation Oral Vaccine
Shelf‐life 3 years
Storage Cold chain (+2 – +8 °C)
Stability at ambient temperature 1 month at 37 °C
Estimated price per dose $1.20
OUR DRUG

10. Toxin Co-regulated
Pilus A (TCPA)
Toxin GM1 binding sites
(GM1 in blue)
Toxin bound to
intestinal cell surface
STRUCTURE

11. Mode of Action of Cholera

12. DISCOVERY
1.Target identification:
•improved cholera vaccine: contains both toxoid and TcpA subunit
•Cell line: New Zealand white rabbit B cell-myeloma cell hybridoma
2.Compound screening
•Infect V. Cholerae with recombinant CTX virus and isolated toxoid produced
•Isolate genetic sequence for the TcpA subunit and have another bacteria to mass produce it
•Transfect hybridoma rabbit B cell-myeloma cell with antigens for antibody production
•Purify antibodies, measure amount of titer produced
•Mix solutions of antibodies and antigens (one vial for TcpA and another for toxoid) to test for
specific binding
3.Lead identification
Mass spectrometry
•Rabbit antibody weight is expected to be ~50 kDa
•Antigen (TCP) weight is expected to be ~20.5 kDa
•Toxoid weight is expected to be 11-23 kDa
Other methods include immunohistochemical assay, immuno co-precipitation and ELISA

13. PRE-CLINICAL STUDIES
Experiment: test safety
and toxicology
Animal model: Five- to
six-week-old Sprague
Dawley rats
Type of studies:
randomized,
vaccine/placebo controlled
Experiment: determine
pharmacokinetic properties
of the drug
Animal model: Five- to six-
week-old Sprague Dawley
rats
Type of studies:
randomized,vaccine/placebo
Experiment: test
developmental and
reproductive toxicity
Animal model: New
Zealand white rabbits
Type of study:
randomized,
vaccine/placebo
Experiment: test efficacy
Animal model: New
Zealand white rabbits
Type of studies:
randomized,
vaccine/placebo
Experiment: determine
pharmacokinetic properties
of the drug
Animal model: 8 to 14
weeks old BALBc mice
Type of study:
randomized, multi-dose

14. CLINICAL
TRIALS
Name of the trials: ACTS studies (Anti-Cholera Toxoid and Anti-Cholera Sub-
unit Studies)
Name of the drug: Cholonex
Category: OCV- Oral Cholera Vaccine; Anti-Cholera Toxoid and Anti-Cholera
sub-unit
Patient Restrictions:
• Not to be given to Pregnant women
• Not to be tested in immunocompromised patients
• Not to be given to patients undergoing any immunosuppressant
therapies or any therapy involving anti-toxics or anti-biotics

15. 50
VOLUNTEE
RS
HEALTHY
VOLUNTEER
S
BLOOD IS
TESTED FOR
ANTI-BODY
TITER
ALL
SEXES
NO
MASKING
Up to
$1M
8 Months to
1 year
NON-
RANDOMISED
US
A
18 to
49
years
Official title: A phase I non-randomized study to evaluate
the Safety and Immunogenicity of the Live Oral Cholera
Vaccine Candidate CHOLONEX

16. 80
VOLUNTEE
RS
HEALTHY
VOLUNTEER
S
BLOOD IS
TESTED FOR
ANTI-BODY
TITER
ALL
SEXES
No-
maskin
g
MAX
TOLERATED
DOSE
18
to
49
year
s
NON-
RANDOMISED
Up to
2 years
Around
$5M
USA,
Africa,
South
Asia
Official title: A phase II study to test the efficacy and immunogenicity
of the Live Oral Cholera Vaccine Candidate Cholonex

17. 2000
VOLUNTEER
S
HEALTHY
VOLUNTEE
RS
SHANCHOL
ALL
SEXES
Around
$20M
Africa
18
to
49
year
s
South
Asia
USA
Randomised,
Double
blinded
Official title : Phase III clinical trial of patients to generate
statistically significant data about safety, efficacy and the
overall benefit-risk relationship of the drug.

18. Objective: to determine the incidence
of any adverse reactions.
• Strict pharmacovigilance
• Collection of post marketing data
• Creating a database for
pharmacogenomics of our drug.
• Submission of periodic reports to
FDA

19. INTELLECTUA
L PROPERTY
• We are aiming a patent related
to the area of immunotherapy.
• Targeting for 20 years of
exclusivity period.
• We are trying to the protect the
cell line and our product.
• Our plan is to file the patent
while we are in mid-way of
preclinical studies
• We are to hoping for it to be
granted by USPTO, 5 years after
we file it.

20. MANUFACTURING: CELL LINE
DEVELOPMENT
1. Create V. Cholerae (El Tor biotype) colony
2. Engenere plasmid coding for the toxin
3. Remove the nucleic acid sequence coding for the GM1 binding site
4. Transfect V. Cholerae with the genetically engineered plasmid
5. Select and culture the transfected bacteria
6. Screen for bacteria with highest production rates
7. Freeze recombinant bacterial cells for the research cell bank and prepare the
master and development cell banks

21. MANUFACTURING: VACCINE PRODUCTION
• Outsourcing to BHARAT BIOTECHS in India
• Generation of the antigen following GMP standards
• Product purification: Column chromatography
• Addition of other components: adjuvant
• Labeling and packing: industrial refrigerated containers
• Distribution

22. DRUG
PRICING
Cost of developing CHOLONEX
vaccine
•Estimated cost of manufacturing in
India $50 million
•Estimated cost of
commercialization in Countries
targeted: $60 million
•Overall cost we need is estimated
to be around $140 million

23. SUMMARY
We are the first US based company to create an
oral vaccine that is specially meant for the aid
of developing countries against Cholera.
We are the first ones to target the specific
sub-unit within the V.Cholarae and the toxin it
produces to effectively disrupt the entire
mechanism of action.

24. REFERENCES
• https://books.google.com/books?id=UVta7XZJHKYC&pg=PA32&lpg
=PA32&dq=t4p+conserve&source=bl&ots=E9QH8Ad63w&sig=ACfU
3U0Vy7kLQHtJCJRzmY_kb-
qrnzUFgw&hl=en&sa=X&ved=2ahUKEwjdxP6O55fmAhXKqZ4KHYs
XClwQ6AEwAHoECAsQAQ#v=onepage&q=conservation&f=false
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC517590/
• https://www.nature.com/articles/s41579-019-0195-4
• https://www.ncbi.nlm.nih.gov/pubmed/15271902
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5518734/
• https://www.nature.com/articles/s41579-019-0195-4
• https://www.niaid.nih.gov/research/vaccine-adjuvants
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4142487/
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5614415/

25. THANK YOU.
ANY QUESTIONS?
