1. Dr. Khaled Al-Qaoud
R&D manager
Jordan Company for Monoclonal Antibody Production
(MonoJo)
www.monojo.com.jo
Jordan Pharmaceutical Association (JPA)
in Collaboration with
Jordan Company for Antibody Production
(MONOJO)
27. Feb, 2013
Biopharmaceuticals: A
New Era in Human Health
3. MILESTONES IN
PHARMACEUTICAL INDUSTRY
www.monojo.com.jo
1895 Aspirin (Bayer)
1930 Sulphanilamide (Sulpha drugs)
1930 Large Scale Production of Insulin
1922 Discovery of Insulin
1940 Penicillin Production
1953 Structure of DNA
1973 Genetic Engineering
1975 Monoclonal Antibodies
AGE OF BIOPHARMACEUTICALS
>10,000 COMPANIES
100 INTERNATIONAL PLAYERS
5000 PHARMACEUTICAL PRODUCTS
$1.3T 2014
TOP 5 DRUGS ARE BIOPHARMACEUTICALS
50% OF TOP 100 ARE BIOPHARMACEUTICA
1980 Recombinant Insulin
5. PROBABILITY OF SUCCESS TO MARKET FOR CHEMICAL
ENTITIES AND BIOTECHNOLOGY PRODUCTS
www.monojo.com.jo
0%
10%
20%
30%
40%
50%
60%
70%
80%
First human dose to
market
First patient dose to
market
First pivotal dose to
market
Submission to market
Probabilityofsuccesstomarket
BiotechChemical entities
90%
100%
Based on 485 new chemical entities and 61 biotechnology products in development within 32 companies.
NAS probability of success to market. Based on NASs entering a phase between 1997 - 1999 where a
decision was made by 31st December 2002
6. Fall in Chemical
Drugs launched
www.monojo.com.jo
2003 LOWEST NUMBER OF NCE LAUNCHES IN 10 YEARS*
NumberofNCEslaunched
42 42 43
51
45 45
41
37 36
30 31
0
10
20
30
40
50
60
1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004
30
33
27
26
28
2005 2006 2007 2008 2009
7. Top 10 Drug Products by Sales
in 2008
www.monojo.com.jo
8. Future Drug Sales Predictions Highlight Importance of
Follow-on Biologics Legislation
Top 10 Drug Products by Sales in 2014
www.monojo.com.jo
9. DRUG DISCOVERY PROCESS AND
TIMEFRAMES
www.monojo.com.jo
DRUG
DISCOVERY
PRE-CLINICAL CLINICAL TRIALS
FDA
REVIEW
LARGE SCALE
MANUFACTURING
PHASE IV
PHASE I
20-100
VOLUNTEERS
PHASE III
1000-5000
VOLUNTEERS
PHASE II
100-500
VOLUNTEERS
FDA
APPROVED
DRUG
5 1.5 6 2 2
10,000 250 5
1
TIME
(YEARS)
COMPOUNDS
IND
NDA
SYNTHESIS/
PURIFICATION
ANIMAL
TESTING
0-1000mg 1-1000g 1-1000kgAMOUNTS
$ $800-1,200M
13. BIOPHARMACEUTICALS OUTGREW TOTAL PHARMA
SALES OVER LAST 10 YEARS, DRIVEN BY US MARKET
www.monojo.com.joYear
0
10
20
30
40
50
60
1994
1995
1996
1997
1998
1999
2000
2001
2002
2003
2004
2005
2007
2006
2008
80
70
4.5%
9%
US 54%
Total
Biopharm
sales [$B]
14. What is a biological medicine ?
www.monojo.com.jo
• Is a medicine whose active
substance is made by or derived
from a living organism (through
Biotechnology as genetic
engineering).
• Example such as : insulin can be
produced by an E. coli bacteria.
15. MILESTONES IN
PHARMACEUTICAL INDUSTRY
www.monojo.com.jo
1895 Aspirin (Bayer)
1930 Sulphanilamide (Sulpha drugs)
1930 Large Scale Production of Insulin
1922 Discovery of Insulin
1940 Penicillin Production
1953 Structure of DNA
1973 Genetic Engineering
1975 Monoclonal Antibodies
AGE OF BIOPHARMACEUTICALS
>10,000 COMPANIES
100 INTERNATIONAL PLAYERS
5000 PHARMACEUTICAL PRODUCTS
$1.3T 2014
TOP 5 DRUGS ARE BIOPHARMACEUTICALS
50% OF TOP 100 ARE BIOPHARMACEUTICA
1980 Recombinant Insulin
16. The Pharmacy of the Future as envisaged by a customer of Behring
22. THE AGE OF
BIOPHARMACEUTICALS
www.monojo.com.jo
RECOMBINANT DNA TECHNOLOGY HAS
4 MAJOR IMPACTS:
SOURCE AVAILABILITY
PRODUCT SAFETY
ALTERNATIVE TO DIRECT EXTRACTION FROM
INAPPROPRIATE/DANGEROUS SOURCE
MATERIAL
FACILITATES GENERATION OF IMPROVED
THERAPEUTIC DISPLAYING SOME CLINICAL
ADVANTAGE OVER NATIVE PROTEIN PRODUCT
23. www.monojo.com.jo
General Scheme for Biopharmaceutical Bulk Drug
Substance Processes
Intracellular
(microbial
fermentation)
Bulk Formulation
PurificationPurification
Isolation/Recovery Isolation/Recovery
Cell Disruption/Refold
Cell Harvesting Cell Removal
Bioreactor Conversion
Bulk Formulation
Working Cell Bank
Extracellular
(microbial
fermentation
and mammalian
cell culture)
“Downstream”
Process
“Upstream”
Process
24. www.monojo.com.jo
....ATG Human Gene
Sequence STOP...
Cloning into
DNA Vector
Transfer into Host Cell
Expression
e.g., bacterial or mammalian cell
DNA Vector
ATG
Fermentation
Stop
Downstream
Purification
Biologic manufacturing is complex
Biosimilars will always be different from the original
Courtesy of Georg-B. Kresse
Roche Pharma Research
26. ADVANTAGES AND DISADVANTAGES OF
EXPRESSION IN E. coli
www.monojo.com.jo
WELL CHARACTERIZED MODEL
FOR PROKARYOTIC GENETICS
HIGH LEVELS OF EXPRESSION
(>45% TOTAL CELLULAR PROTEIN)
E. coli CELLS GROW ON SIMPLE/
INEXPENSIVE MEDIA
FERMENTATION TECHNOLOGY
WELL ESTABLISHED
HETEROLOGOUS PROTEINS
ACCUMULATE INTRACELLULARLY
INABILITY TO UNDERTAKE
POST-TRANSLATIONAL MODIFICATIONS
LIPOPOLYSACCHARIDE (ENDOTOXIN)
28. PROTEINS PRODUCED IN MILK OF
TRANSGENIC ANIMALS
www.monojo.com.jo
Protein
tPA
Interleukin-2
Factor VIII
Factor IX
a1-Antitrypsin
Fibrinogen
Erythropoietin
Antithrombin III
Human a-Lactalbumin
Insulin-like Growth Factor
Protein C
Growth Hormone
Animal Species
Goat
Rabbit
Pig
Sheep
Goat
Sheep
Rabbit
Goat
Cow
Rabbit
Pig
Rabbit
Expression levels in milk (g/l)
6
0.0005
0.003
1
20
5
0.05
14
2.5
1
1
0.05
29. PROTEINS PRODUCED IN TRANSGENIC
PLANTS
www.monojo.com.jo
Protein
Erythropoietin
HSA
Glucocerebrosidase
Interferon-a
Interferon-b
GM-CSF
Hirudin
Hepatitis B Antigen
Antibodes/Fragments
Expressed in
Tobacco
Potato
Tobacco
Rice
Tobacco
Tobacco
Canola
Tobacco
Tobacco
Expression levels
0.003% total soluble plant protein
0.02% soluble leaf protein
0.1% leaf weight
-
0.00002% fresh weight
250 ng/ml extract
1% seed weight
0.007% soluble leaf protein
Various
COST OF PLANT CULTIVATION IS LOW
HARVEST EQUIPMENT IS INEXPENSIVE/WELL ESTABLISHED
EASE OF SCALE-UP
PROTEINS EXPRESSED IN SEEDS ARE STABLE FOR YEARS
PLANT-BASED SYSTEMS ARE FREE OF HUMAN PATHOGENS
VARIABLE/LOW EXPRESSION LEVELS
POTENTIAL OCCURRENCE OF GENE SILENCING
(SEQUENCE SPECIFIC mRNA DEGRADATION)
NON-NATIVE HUMAN GLYCOSYLATION PATTERN
SEASONAL/GEOGRAPHICAL NATURE OF PLANT GROWTH
32. www.monojo.com.jo
The problem of formulating
proteins
• Very large and unstable molecules
• Structure is held together
by weak, non-covalent forces
• Easily destroyed even
under relatively mild
storage/handling conditions
+H3N
Amino end
Amino acid
subunits
helix
CH2
CH
O
H
O
CHO
CH2
CH2 NH3
+ C-O CH2
O
CH2SSCH2
CH
CH3
CH3
H3C
H3C
Hydrophobic
interactions and
van der Waals
interactions
Polypeptide
backbone
Hyrdogen
bond
Ionic bond
CH2
Disulfide bridge
33. www.monojo.com.jo
Why are biopharmaceuticals different?
• High molecular weight
• Complex three-dimensional structure
• Complex manufacturing process
• Produced by living organisms; therefore often
heterogeneous
• Difficult to characterize completely by physico-
chemical analytical methods or bioassays
• Dependence of biological activity on reproducibility
of the production process, in-house standards
• Inherent risk of immunogenicity
37. Evolution of Therapeutic
Antibodies; 3 Main Eras
www.monojo.com.jo
Behring Era
Horse Sera
Kohler & Melstein Era
Mouse Monoclonals
Genetic Engineering Era
Humanized & Human
39. www.monojo.com.jo
Source of antibodies on the market
(according to a “market analyst”)
Source Antigenicity % Human
Mouse +++ 0 %
Chimeric + 60-70%
Humanised - > 90%
Human - 100%
47. What is a Biosimilar?
Similar product if compared to an
original but is made according to
Different process:
different construct, host, cell line,
protocol and/or purification steps
additional pre-clinical tests or
clinical trials required to show
similarity
www.monojo.com.jo
48. Driving forces for biosimilars
(Follow-on Biologics)
• The patents of several biopharmaceuticals
have expired or they are about to expire
• Biologics responsible for 20 Billions USD
annual sales will go off patent by 2015
• Pressure to reduce healthcare expenditure
and increase patient access to treatment
will drive the development of cheaper
biosimilars
www.monojo.com.jo
49. www.monojo.com.jo
Biosimilars: Constraints & Drivers
• Regulatory
– No clear cut regulatory
• Technological
– Manufacturing complexity
– High setup & manufacturing
costs
– Higher cost to prove
comparability
• Market Factors
– Next generation biologics
– Limited discounting ability
(discounting in the range 25%
to 30%)
– Gaining acceptance by
clinicians (brand based
competition)
• Biologics segment size & growth
– Biologics performance exceeds
that of overall market
– Biologics worth $18 bio. Will go
off patent by 2011
• Increasing pressure to reduce
healthcare expenditure
– Evolving regulatory mechanisms
to recognise biosimilars (EU,
Australia)
• Semi/unregulated markets as launch
pad
CONSTRAINTS DRIVERS
51. Data needed to prove similarity
of follow-on biologics
• Analytic studies showing product is highly
similar
• Animal studies (including toxicity)
• Clinical Studies showing safety, purity, and
efficacy including immunogenicity
www.monojo.com.jo
52. The process is complex:
Upstream and downstream
www.monojo.com.jo
53. www.monojo.com.jo
....ATG Human Gene
Sequence STOP...
Cloning into
DNA Vector
Transfer into Host Cell
Expression
e.g., bacterial or mammalian cell
DNA Vector
ATG
Fermentation
Stop
Downstream
Purification
(Probably) a different
DNA vector
A different
fermentation
process
A different
downstreaming
protocol
Different
in-process controls
Maybe the same
gene sequence
A different
recombinant
production cell
Biologic manufacturing is complex
Biosimilars will always be different from the original
Courtesy of Georg-B. Kresse
Roche Pharma Research
55. Similarity and comparability are two distinct
concepts
Only quality data combined with preclinical and clinical experience
provide the full picture
www.monojo.com.joAdapted from Koslowski S, Swann P. Advanced Drug Delivery Reviews 2006; 58: 707–722
In-process
controls
End-product
controls
Release tests
Extended
characterization
Process
56. www.monojo.com.jo 56
Sickle-Cell Disease: A Simple Change
in Primary Structure
• Sickle-cell disease
– Results from a single amino acid
substitution in the protein hemoglobin
57. www.monojo.com.jo
Fibers of abnormal
hemoglobin
deform cell into
sickle shape.
Primary
structure
Secondary
and tertiary
structures
Quaternary
structure
Hemoglobin A
10 m 10 m
Primary
structure
Secondary
and tertiary
structures
Quaternary
structure
Red blood
cell shape
Hemoglobin S
subunit subunit
1 2 3 4 5 6 7 3 4 5 6 721
Normal
hemoglobin
Sickle-cell
hemoglobin
. . .. . . Exposed
hydrophobic
region
Val ThrHis Leu Pro Glul Glu Val His Leu Thr Pro Val Glu
58. ICH Q5E Document on
Comparability
• Provides a harmonized approach for determining comparability
however:
– Requirements for data are not consistent or transparent
• Glycosylation, N-terminal heterogeneity
• Process data
• Impurity data
• Non-clinical safety data
• pK/pD data and additional clinical data
– Application to investigational and approved products is not clear:
59. The Degree of Any Manufacturing Change
Determines How Much Data Will Be Required
59
Change filter
supplier
Move
equipment to
different part
of facility
Lower risk changes
Commonly implemented
Minimal Data Required
(Analytical Testing)
High risk changes
less common
Maximal Data Required
(incl. Clinical Testing)
Move
manufacturing
to new facility
Scale up
manufacturing
DEGREE OF MANUFACTURING CHANGE
INCREASED RISK
New cell line
Change in process
technology
Formulation change
BiosimilarsManufacturing Changes
Commonly Implemented
60. EMEA Approach for Biosimilar Medicines:
Guideline on Similar Biological Medicinal Products
(CHMP/437/04)
• Overall Approach
– Similar biological medicinal products are not generic medicinal
products
– Comparability studies need to demonstrate the similar nature in
terms of quality, safety, and efficacy
• Biosimilars will be different from the reference
– It is not expected that the quality attributes in the biosimilar
and reference product will be identical
– The biosimilar product may exhibit a different safety profile (in
terms of nature, seriousness, or incidence of adverse reactions)
61. Overview Of EMEA Guidelines
Guideline on Similar Biological Medicinal Products
Guideline on Similar Biological Medicinal Products
Containing Biotechnology-Derived Proteins as Active
Substance: Quality Issues
Overarching
Quality
Annexes
Nonclinical
& Clinical
Recombinant
Human
Erythropoietin
Recombinant
Human
G-CSF
Recombinant
Human
Insulin
Recombinant
Human Growth
Hormone
General:
Applies to all
Biosimilars
Specific:
Product data
requirements
Guideline on Similar Biological Medicinal Products
Containing Biotechnology-Derived Proteins as Active
Substance: Nonclinical & Clinical Issues
Nonclinical
& Clinical
TOPIC TITLE APPLICATION
64. high degree of isoform variability in
rEPO products
www.monojo.com.jo
each of the two batches of Huan Er Bo, indicating
differences in the manufacturing process of the
product within the same company.
65. www.monojo.com.jo
Even what looks the same may be different
Absence of Evidence is not Evidence of Absence
IEF pattern and sialic
acid content of the two
EPO isoform preps are
very similar – but
bioactivity is different
huEPO-(1) huEPO-(2)
Isoform 2 Isoform 2
Sialic acid14.0 14.2
in vivo
activity
(U/mg)
226,000 400,000
1
8
7
6
5
4
3
2
1
8
7
6
5
4
3
2
Courtesy of Georg-B. Kresse
Roche Pharma Research
The carbohydrate
structures of the
two EPO isoforms
differ
Carbohydrate
structures
huEPO-(1)
(isoform 2)
huEPO-(2)
(isoform 2)
67. www.monojo.com.jo
Characteristic Method IPC DS DP ST
Methods
Identity Western Blot X X X X
Titer ELISA X
Standard Analytical Methods
In-Process Cell
Culture
Cell count, viability, pH, C02,
glucose X
Appearance Visual examination X X X
pH Potentiometric X X X X
Identity
Peptide Map X X X
Western Blot X X X X
Quantity
Analytical Protein A X X X X
ELISA X
Purity
SDS-PAGE X X X X
RP-HPLC X X X X
Impurities
Host Cell DNA - qPCR X X
Host Cell Protein - ELISA X
Residual Protein A -ELISA X
Protein Content
A280 X X X
BCA X
Endotoxin Kinetic Ph. Eur. X X X X
Bioburden Culture Ph. Eur. X X X X
Mycopiasma Unprocessed bulk harvest X
Viral testing Unprocessed bulk harvest X
Excipients to be determined X
IPC — In Process Control testing DS — Drug Substance release testing
DP — Drug Product release testing ST — Stability testing
70. Monoclonal antibodies
Structurally much more complex than other proteins
• Complexity: considerably more complex than
currently developed biosimilars
• Biological activity: glycosylation patterns are
critical - small differences correlate to changes in
biological activity
• Predictability: multi-functionality (both binding and
immune effector functions) coupled with an
often limited understanding of structure-
function relationship will limit predictability of
in vitro data
• Extrapolation: complexity and diversity of the mechanisms
of action will be of particular challenge for
indications and line extensions
www.monojo.com.jo
72. Reditux (anti CD20): an intended
copy, not a proven biosimilar
• Same amino acid sequence
• Host cell protein content much
higher
• Content of aggregates not
comparable
• Glycosylation not comparable
• Effector function not comparable
• Charge distribution not comparable
• Clinical (PK/PD) published data - 17
patients
www.monojo.com.jo
Different manufacturing means:
• Different drug
• Different safety/efficacy profile?
73. Changes in glycosylation of
antibody will affect Potency
@Potency can be affected if the molecule:
Has Fc function
Has glycosylation in the Fab region
@Effect on PK highly dependent on
sugar moieties
High mannose can clear quicker in vivo
@Bioavailability depends on any charge
differences
@Not easy to control from lot to lot
www.monojo.com.jo
75. Bio-better
• intended to be better or superior to the
innovator product with marked
differences in:
– clinical efficacy
– safety
– and/or convenience
• should go through the full development
and approval process
www.monojo.com.jo
76. More details
• Modified by protein or glyco-
engineering
• efficacious, require a lower dosing
frequency
• most critically, reduce the risk of
immunogenicity
• Even better, they have lower early-stage
R&D costs
www.monojo.com.jo
77. Why going to bio-better
• modifying an existing therapeutic
protein is significantly easier and less
risky than developing a new one
www.monojo.com.jo
78. Biobetter in monoclonal therapeutics
Adding benefits to key medicines; developing better ones
www.monojo.com.jo
79. T-DM1 as an example for next generation
antibodies
www.monojo.com.jo
80. Follow-on Biologics will play a role in
shaping the future of Pharma industry
Main Future Big Players in Follow-on Biologics:
• Teva (large generics manufacturer) partnered
Lunza Group
• Sandoz, generics arm of Novartis: increased
capacity in Bio-manufacturing to ramp up its
effort
• Bioventures of Merck: established in 2008 for
the development of follow-on Biologics
• Pfizer: testing follow-on version of Enbrel (in
phase 2 clinical trials)
www.monojo.com.jo
83. MidPharma-ISU ABXIS
Clotinab®
the first biosimilar of REOPRO® (Centocor)
• is a Fab fragment of a therapeutic
antibody for GP IIb/IIIa receptor (human
platelets)
• inhibits platelet aggregation.
• used for prevention of cardiac ischemic
complications
www.monojo.com.jo
