Vaccination and developed of new vaccines

World Health Organisation

"Nearly nine million children under 14 years of age

die every year from infectious disease. And at least a
third of them could be saved if existing vaccines were
more widely used, but the rest only if suitable
new vaccines were developed..."

- the World Health Organization

http://www.who.int/gpv-news/
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Global Infectious diseases
Global Mortality
Figure 1. Infectious diseases and global mortality, 1995 (all ages)

Mortality preventable with Hepatitis B 1.1 million

available vaccines
Measles >1 million
Neonatal tetanus Tuberculosis 3.1
500,000 million
Pertussis 355,000 Malaria and other
vector borne diseases
2.2 million

Acute respiratory
infections 4.4 million HIV/AIDS >1 million

Others 1.2 million Diarrheal diseases 3.1

million
Roundworm and
hookworm 165,000

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Vaccine development

1950- Most vaccines produced used traditional technology for past 40 yrs
1982 Plasma derived Hepatitis B Vaccine
1985 Hib was licensed by Praxis, Rochester, NY
1986 Recombinant Hepatitis B Vaccine - Merck
Mid 80- New vaccines from advances in Mol. Biol., Immunology, production tech
90s and drug delivery system. 14 new or improved vaccines developed
1997 International Vaccine Institute, Seoul, S. Korea
1999 AIDS vaccine finished phase III clinical trial in USA, started trial in Asia
98-2000 8 new vaccines expected, 4 potentials to prevent rotavirus, enterotoxic
Escherichia coli (ETEC) or cholera, pneumococcal pneumonia & meningococcal diseases

Vaccine Market
1985 US$ 300m
1996 US$ 4,000m
2000 US$ 7,000m

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Vaccine development
New Vaccine Strategies

Purified (Subunits) Antigens vaccine e.g. Hepatitis B, Haemophillus

influenza type b, RSV, Rotavirus, foot-and-mouth disease

Conjugate vaccines e.g. meningitis, pneumonia

Recombinant antigen vaccines e.g. Hepatitis B, malaria

Synthetic peptide vaccines e.g . parasites (malaria),

bacteria (diphtheria and cholera) toxins, viruses (HIV)

Recombinant vector vaccines e.g. investigated for hepatitis B virus,

herpes simplex, influenza virus, and HIV

DNA vaccines e.g. purified preparation of plasmid DNA vaccines,

especially for immune deficient recipients

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The Vaccine Market
The Vaccine Market
The Market Leader
SmithKline Beecham 33% Market
Vaccines Sales 1996 US$ 996m
HBV, Enegerix-B US$ 568m
HAV, Harvix(1996 in China) US$ 256m
Infanrix (DPT)

Other Large vaccine companies:

Pasteur Merieux Connaught - vaccines for 18 diseases
Merck - Vaccines for chickenpox, HAV,
Haemophilus influenzae type b & HBV
- 50%-owned joint venture with Pasteur Mérieux
Bristol Myers - two experimental cancer vaccines
CEL-SCI Corporation - improved HGP-30AIDS vaccine
RIVM Holland - National Vaccination Programme, transferring
knowledge to developing countries

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The Vaccine Market
The Vaccine Market

The Market Leader

SmithKline Beecham 33% Market
Vaccines Sales 1996 US$ 996m
HBV, Enegerix-B US$ 568m
HAV, Harvix(1996 in China) US$ 256m
Infanrix (DPT)

Other Large vaccine companies:

Pasteur Merieux Connaught - vaccines for 18 diseases
Merck - Vaccines for chickenpox, HAV,
Haemophilus influenzae type b & HBV
- 50%-owned joint venture with Pasteur Mérieux
Bristol Myers - two experimental cancer vaccines
CEL-SCI Corporation - improved HGP-30AIDS vaccine
RIVM Holland - National Vaccination Programme, transferring
knowledge to developing countries

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The Vaccine Market
The Vaccine Market

The Market Leader

SmithKline Beecham 33% Market
Vaccines Sales 1996 US$ 996m
HBV, Enegerix-B US$ 568m
HAV, Harvix(1996 in China) US$ 256m
Infanrix (DPT)

Other Large vaccine companies:

Pasteur Merieux Connaught - vaccines for 18 diseases
Merck - Vaccines for chickenpox, HAV,
Haemophilus influenzae type b & HBV
- 50%-owned joint venture with Pasteur Mérieux
Bristol Myers - two experimental cancer vaccines
CEL-SCI Corporation - improved HGP-30AIDS vaccine
RIVM Holland - National Vaccination Programme, transferring
knowledge to developing countries

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Vaccine Report
Vaccine Development
ccines Product in Development
mpany Plan/R&D Preclinical Phase I Phase II Phase IIIa Phase IIIb Marketed Annual Market
Sales share
ithkline EBV Campylobacter MMR-Varicella DTPaHepBHiB DTPa, Harvix US$996m 33%
cham Otitis media / Helicobacter Improved DTPaIPVHepBHib DTPa Hib (Hepatitis A) in 1996
MeningitisA/C pylori Hepatitis A/A&B Enegerix
ccines Acellular Pertussis Hepatitis A&B (Hepatitis B)
RSV comb. adults (Twinrix) Infanrix
Lyme Disease MMR , Varicella (Varilix) (diphtheria,
HSV, Cholera DTPwHepB tetanus and
Enterotoxigenic E. Coli pertussis
Salmonella Thypi vaccine)
DTPaHepB
DTPaIPVHiB
ithkline Melanoma Human Papilloma
cham Herpes Simplex Virus
munothera Hepatitis B
tic
cines
teur Pox virus Combination Lyme around 20%
rieux vector Pediatric 600 mUSD
nnaught construct Meningitis
(conjugate)
Pneumococcal
(conjugate)
Influenza
(improved)
rck Sharpe 663 MUSD 22%
Dohme

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Vaccine development
Targets for new vaccines

Major targets for new or improved vaccines

Amebiasis Filariasis Leprosy Schistosomiasis

Ascariasis Giardiasis Onchocerciasis Shigella
Cancer Gonorrhea - skin disease Streptococus mutants
Chlamydia Hantaan Virus - river blindness Syphilis
Coccidiomycosis Herpes Virus Respiratory infections Trichuriasis
Cytomeglovirus Hepatitis C - N.Meningococcus B Trypanosomiasis
Dengue fever HIV / AIDS - Parainfluenza - African type
Malaria Human Papilloma - Resp. Syncytial Virus - S. American type
Epstein-Barr Virus Japanese Encephalitis - S. pneumoniae Tuberculosis
Fertility Control Leismaiasis Rotavirus

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Vaccine development
Vs diseases in developing countries
Table 2. Summary of the status of vaccine development and use against the major
diseases in developing countries.

Disease Groups and Vaccines Status of Vaccine Development Widely-used?

Acute Haemohilus Licensed vaccine widely used in No
Respiratory Influenza b many developed countries
Infections Streptococcus Conjugate vaccines effective in No
Pneumoniae infants are in advanced stage of
development
Neisseria Conjugate vaccines effective in No
meningitidis infants are in advanced stage of
development
Respiratory Vaccine in clinical trials No
syncitial virus
Enteric Cholera New vaccines with increased No
diseases effectiveness are in clinical trials
Enterotoxigenic Vaccine candidates in clinical trials No
E. Coli
Rotavirus New vaccine effective against No
severe form of disease is in final
stages of licensure
Shigella Vaccines are in early stages of No
development
Typhoid Two licensed vaccines, one with No
variable efficacy and the other not
fully effective in infants
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Vaccine technology
Vs diseases in developing countries

Disease Groups and Vaccines Status of Vaccine Widely-

Development used?
Tuberculosis Current vaccine has Yes
variable effectiveness
Malaria and Dengue Candidate vaccine in early No
other vector- clinical trials
borne Japanese Several vaccines available Yes
encephalitis but with various drawbacks
Malaria Candidate vaccines in early No
clinical trials
Hepatitis B Hepatitis B Vaccines licensed Coverag
vaccine e of new
born still
limited
HBV combined Vaccine licensed No
with other
vaccines
HIV/AIDS Candidate vaccines in early No
stages of development
Measles Vaccines licensed Yes
Neonatal Tetanus Vaccines licensed Yes
Pertussis Vaccines licensed Yes
Roundworm and Hookworm Vaccine candidates in No
animal studies

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Vaccine Market

‘We help people to make better vaccines’

Attenuated/Inactivated whole organism Microcarriers Cytodex, Cytopore (for viral

vaccines). Gel Filtration and Ion Exchange purification
Purified (Subunits) Antigens Microcarriers Cytodex, Cytopore and reactors
Conjugate vaccines Molecular biology and cell biology reagents
Recombinant Antigen Large & lab-scale purification systems and media.
Recombinant Vector Down-stream processing technology & know-how
Electrophoresis - SDS-PAGE
Synthetic peptide vaccines Purification systems and media
DNA vaccines Molecular Biology and Cell Biology System & Reagents
MegaBase, OliogoPilot, OligoProcess, BioProcess

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 Vaccine Production with Microcarriers

Separates cells from secreted

products
Large volume to surface ratio
(less waste problem)
Cheap surface supply/m2
Proven scalable technology
(1000’s of l)
Increased productivity of
functional product Polio vaccine production in 1000L fermentors,
Institute Pasteur Merieux, France

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 Microporous carriers: Cytodex

Lower cell density in the

supernatant Porous
structure
of Cytodex 1
Necessary for adherent cell lines
Gives cell support to build
cytoskeleton, intracellular
organisation
Allow cells to create micro-
environment inside carrier

Vero cells grown on Cytodex 1

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Cytodex production examples

Pasteur Merieux: JEV, Vero Cells up to 2000 l cultures.

Large scale production of vaccine against Japanese encephalitis in cultured cell lines, Patent.

Connaught-Lab: WI-38 and MRC-5 in 50l to 1000 l culture.

In-Vitro (1993), Comparison of large scale serial subcultivation of WI-38 vs MRC-5 on
Microcarriers.

Celltech: human diploid fibroblast up to 4000l on Microcarriers.

Protein-Prod. Biotechnol. (1990), Large scale culture of mammalian cells for production of
therapeutic proteins.

Smith Kline: Vero cells, polio virus, Cytodex 3, cell density up to 14,5 million
/ml, virus titer up to 10 power 10,13 TCID50/ml.
Adv. Anim. Cell-Biol. Technol. Bioprocesses (1989), High density microcarrier culture for
viral vaccine production.

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 Macroporous carriers: Cytopore
Substitution macroporous particle
DEAE (1.0, 1.85 meq/g) throughout
matrix
Much higher surface area-1.1M2
/gram
Give protection from stirrer, spin
filter and air/O2 sparging Porous
structure of
Give higher cell densities Cytopore

Shorter residence time of product at

37°C
Allow cells to grow in 3D rCHO cells grown on Cytopore 1

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 Cytopore production examples

3800
4000

3500 3200

3000

2500
2000
1800
2000
IL-4 ug/12 days
1500

1000

500

0
Cytopore (10% Suspension Cytopore (S.F.) Suspension
CS) (10% CS) (S.F.)

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 Microcarriers in roller bottles

Much higher surface area

Better protection
Give higher cell densities
Economic and simply way to
increased productivity
Three methods
•Adhere to surface of roller ~1.5mg/cm2
•Suspened in culture by siliconization
•Grow to onfluence in roller bottle
Mouse L-cells in roller bottle culture using
cutodex to increase yield, at 8h, 24h, 72h, 96h

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 Vaccine Downstream Purification
Gel filtration method on
Sepharose 4,6FF in BPG
or INdEX Albumin, phenol red, impurities

Manual Process system:

100-400L/hr
Rabies vaccine
Loading upto 10% CV

High degree of
automation, sanitary,
reproducible, simple

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Vaccine Downstream Purification at large scale

BioProcess system at Smith Kleen Beechem

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 1st rDrug Using STL
Approved by FDA
• SKB,a lipoprotein, human vaccine
• Direct capture on STL400 after
high pressure homogenization of
E.coli. Combining 3 steps into 1.
• Further purified on two Sepharose
IEX and Sephacryl in Chromaflow

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