Dennis M. Gross, MS, PhD
CEO & Treasurer
Pennsylvania Drug Discovery Institute
© Dr. Dennis M. Gross 2015
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Definitions: Large molecules
Diff
D
Bi l i
Differences: Drugs vs. Biologics
Vaccines
Case Studies from Production of Vaccines
Development
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Non‐clinical
Clinical
Future of Vaccines and Biologics
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1
Vaccines
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Attenuated: M‐M‐R, Rotavirus, Varicella, Zostavax
Killed or Inactivated: IPV, HepA
Toxoids: Diphtheria; Tetanus;
Sub‐unit: HepB; acellular Pertussis; VLP
Vector‐based vaccine: Canary pox, Adenovirus,
DNA: In research phase
Peptide vaccine: Epitope vaccine, peptide immunogens (HER‐2/neu); 
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Antibodies: MAbs
Cytokines:
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 Lymphokines (IFN, IL); monokines (IL,TNF); Interleukins (IL)
Growth factors: Erythropoietin, VEGF
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Hormones: Steroids (estrogens); Polypeptides (Insulin, endorphins); etc.
Gene Therapy
Stem Cells: Embryonic, Adult
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Drugs
Synthetic, organic compounds
Defined structure, physical & chemical characteristics
chemical characteristics
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Chemical synthesis
Small molecule (±1,000)
Stable
Homogeneous
Biotransformed (metabolized)
Linear dose‐response
Activity not species specific
Acetaminophen Mw = 151 4
2
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Drugs
Synthetic, organic compounds
Defined structure, physical & chemical characteristics
chemical characteristics
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Chemical synthesis
Small molecule (±1,000)
Stable
Homogeneous
Biotransformed (metabolized)
Linear dose‐response
Activity not species specific
Acetaminophen Mw = 151 
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Biologics
Protein‐, carbohydrate‐ or nucleic acid ‐based product
Extracted from living organism
Complex physicochemical structure
Less well‐characterized
Macromolecule (5,000 ‐>1,000,000)
Tertiary structure
Location, extent and type of glycosylation
Heat‐ & shear‐ sensitive
Non‐linear dose‐response
Monoclonal antibody, Mw = ~150,000 5
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Vaccine: A preparation of microorganisms (e.g., bacterium, virus), or their antigenic components administered for the prevention, amelioration or treatment of an infectious disease 
Biologic preparation that elicits immune response with memory
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Generates humoral (antibody) and/or cellular (cytotoxic or killer T‐cell) response to eliminate pathogen or pathogen infected cell
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The immune system is then primed to mount a secondary immune response with strong and immediate protection upon future exposure to ih
di
di
i
f
the pathogen
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1981
Hepatitis B
Ref. Hilleman, Nature Medicine, 4, 504, 1998
Maurice Hilleman
1919 - 2005
Albert Sabin
1906 - 1993
Jonas Salk
1914 - 1995
1923
Diphtheria
Louis Pasteur
1822 - 1895
Edward Jenner
1749 - 1823
1796
Smallpox
1780 1800
1926
Pertussis
1927
Tetanus
1840
1860
1880
1962
Oral Polio
1986
Hepatitis
B
p
(Recombinant)
1963, 1968
Measles
1989
Hib Conjugate
1965
Japanese
encephalitis
1995
Varicella
1967
Mumps
1996
Hepatitis A
1969
Rubella
1996
Hib-Hep B
1971, 1978
MMR
1996
DTacP
1974
Meningococcal
1998
Lyme
1977
1935
1956 Pneumococcal
Yellow Fever Adenovirus 14-valent
1885
Rabies
1820
1955
Polio
1900
1920
1940
1983
Pneumococcal
23-valent
1960
1999
Pneumococcal
conjugate
1980
2000
4
Smallpox
Pertussis
Diphtheria
Polio Vaccines
Hepatitis B vaccine
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Protection from Cholera
Picture courtesy of Wellcome Library, London
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17th century Chinese practice
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Vaccinating with scabs from smallpox patients
smallpox patients
1796 ‐ Edward Jenner
Vaccine made from pus of cowpox patients and milk maids
 Coined the term “vaccination”  First recognized use of a vaccine
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No purification!
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Whooping cough caused by B. pertussis, a small gram negative bacteria
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Vaccine ‐ suspension of killed whole cells of bacteria 
Whole cell pertussis vaccine first licensed in the US in 1914
 Later combined with diphtheria and tetanus toxoids (DTP) in 1942
tetan s to oids (DTP) in 1942
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Lack of purity caused some side effects
 Led to the development of acellular pertussis (subunit vaccine)
Ref. Aunins, Lee and Volkin, in The Biomedical Engineering Handbook, CRC Press, Boca
Raton, 1995
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Respiratory infection caused by C. diphtheriae, a gram‐positive bacteria
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Diphtheria toxin, a polypeptide of 62 kDa MW, is formaldehyde treated to form toxoid
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First protein purified for active immunization
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Purity < 60%
P it 60%
 protein impurities can crosslink with toxoid
Ref. Rappuoli and Pizza in Vaccines, Eds. Perlman, P.
and Wigzell, H., Springer, Heidelberg, 1999
6
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Enders successfully cultivated virus in tissue culture (1949)
Inactivated virus developed by Salk (1955)
Oral polio (Live virus) developed by Sabin (1963) (1963)
Oral polio (Live virus) developed by Sabin
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Less expensive and simple to use
IPV
Ref. Salk, JAMA, 151, 1081, 1953
OPV
Ref. Aunins, in Encyclopedia of Cell Tech., Ed. Spier, Wiley & Sons, NY, 2000
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Research & Development
Upstream Processes
Upstream Processes
Production
Downstream Processes
Test Article
Gene Discovery
Cloning & Transfection
Transformation
Cell Line Development
p
Media Development
Microbial Fermentation
Mammalian Cell Culture
Harvest Cells
Cell Disruption
Product Isolation
Product Purification
Analytical Tests
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7
Research & Development
Upstream Processes
Upstream Processes
Production
Downstream Processes
Product
Gene Discovery
Cloning & Transfection
Transformation
Cell Line Development
p
Media Development
Microbial Fermentation
Mammalian Cell Culture
Harvest Cells
Cell Disruption
Product Isolation
Product Purification
Analytical Tests
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 Small‐scale (<10 L)
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‐ Flasks (T‐flasks, Erlenmeyer, Spinners)
‐ Roller Bottles
 Mid‐scale (10 ‐ 250 L)
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‐ Roller Bottles (automated handling)
‐ Nunc Cell Factories and Costar Cell cubes
‐ “Small” continuous stirred reactors
‐ Wave Bioreactors
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Wave Bioreactors
Large scale (> 250 L) engineered systems.
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‐ suspended cells.
‐ suspended cells on ‘Microcarriers’.
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Large scale (> 250 L) engineered systems.
Images: Copyright Sanofi Pasteur , Amgen,
and New York Times
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Fever and jaundice of 2‐3
weeks duration
Fever and jaundice of 2
3 weeks duration
Transmitted by fecal oral route
In US, spread through day care centers, residential institutions, restaurant food handlers
Endemic in developing countries
Vaccine development undertaken to minimize spread of disease
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Ref. Aboud et al. (Merck & Co. Inc.), PCT WO 94/03589, 1994
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7
8 9
1. LOW MW STANDARDS
2. LYSATE
3. LYSATE + BENZONASE
4. CAPTURE PRODUCT
5. PEG PELLET
6. SOLVENT EXTRACT
7. ION EXCHANGE PRODUCT
8. SIZE EXCLUSION PRODUCT
9. SIZE EXCLUSION PRODUCT
12% GEL WITH 200NG HAV ANTIGEN PER LANE
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Resuspended PEG Pellet Sample
Ion-Exchange Chromatography Product
660kDA Protein
Hepatitis A
Hepatitis A
Aggregate
660kDA
Protein
Aggregate
A 214nm
0
A 214nm
10
20
30
40
0
Solvent Extracted Aqueous Phase Sample
10
20
30
40
Size-Exclusion Chromatography Product
Hepatitis A
Hepatitis A
660kDA
Protein
Aggregate
A 214nm
0
A 214nm
10
20
Minutes
30
40
0
10
20
Minutes
30
40
Hardware: Rainin HPLC system, TosoHaas TSK PW4000xl, 7.8 x 300 mm column
Mobile phase: PBS, 0.32 ml/min, 50
l injection. Detection UV 214 nm
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Pre‐clinical
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Drugs
Toxicology in rodent and something else
Hepatic microsome for CYP enzyme metabolism
Safety pharmacology (CNS, cardiovascular, respiratory studies ICH S7)
Carcinogenicity (Ames Test)
Clinical
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Drug‐drug interactions
Food‐drug interactions for oral administration
Effect on QT/QTc & arrhythmic potential
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Pre‐Clinical
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Biologics
T i l
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l
t i l
Toxicology in relevant animal species
Very difficult learning curve at the moment
Don’t always get NOAEL (No Observed Adverse Event Level)
Clinical
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PK and bioavailability but not metabolism
Concomitant use (vaccines) Off‐target incorporation of material (DNA Vaccines)
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To support entry into clinical trials, where human safety is ultimately evaluated
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Rare toxicities, or potential effects of sub‐populations often only addressable in humans
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Maximize the benefit‐to‐risk of vaccine development
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Determine a safe dose
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Identify any potential or unknown toxicities, target organs
 Broad measures ‐> unpredictable toxicity
 Specific assays ‐> key theoretical concerns
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Inherent toxicity of the vaccine
Inherent toxicity of the vaccine
Toxicity of impurities/contaminants
Toxicity due to interaction of components
Toxicity linked to the immune response T i it li k d t th i
induced
Unknown unknowns (Black Swans)
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Route of administration (ROA) and dose should correspond to clinically intended ROA and dose(s), e.g., oral, SC, IM, Nasal
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Total number of doses equal to or exceed number of clinically administered doses
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[“N plus 1”]
Episodic dosing, e.g., weeks between doses
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“Relevant” animal species
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An animal species susceptible to respond to the test article activity, e.g., development of an immune response after vaccination
 Ideally, species should be sensitive to the pathogenic organism or toxin
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One relevant animal species in general is sufficient
 Exceptions on a case‐by‐case
 Non‐human primates not generally necessary
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Group size dependent on the animal model
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IND/
CTA
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NDA/
WMA/
BLA 
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Phase I – First‐into‐Human
 20‐100 normal healthy volunteers
Determine safety and tolerability
 Determine safety and tolerability
Phase IIa/IIb – Therapeutic Exploration
 100‐300 patients
 Evaluate efficacy
 Determine dose range and adverse events (AEs)
Phase III – Therapeutic Confirmation
 1,000 to 15,000 patients
 Verify efficacy, monitor AEs and effect in usual therapy and special populations
Phase IV – Post‐Approval
 Postmarketing surveillance
 Monitor long term risks and benefits, outcomes and pharmacoeconomics
Phase V
 Postmarketing studies (Potentially thousands of patients)
 New indications: “Megatrials”
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Sources: Clemens et al. JAMA 1996;275:390‐7
Levine MM. British Medical Bulletin 2002;62:1‐13
Promising vaccine candidate
IND
Phase I: Well‐tolerated and immunogenic in healthy adults?
No
Yes
Phase II: Well‐tolerated and immunogenic in the target population?
Yes
No
Phase III: Well‐tolerated and efficacious in the target population?
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Phase III: Well‐tolerated and efficacious in the target population?
No
Subset assessed for immunogenicity
y
y
p
Antibody level may link with protection = CORRELATE OF PROTECTION
Yes
Phase IV: Assess effectiveness
Sources: VacciNews 2002;1(6):1‐4 Clemens et al. JAMA 1996;275:390‐7
Levine MM. British Medical Bulletin 2002;62:1‐13
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trial provides a more direct
An Efficacy trial provides a more direct
measurement of protection
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Efficacy‐ the protective effect of a vaccine against the target disease
Protective Efficacy =
incidence of disease in vaccinees
1- incidence
of disease in nonvaccinees X 100%
Sources: VacciNews 2002;1(6):1-4 Clemens et al. JAMA 1996;275:390-7
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IND/
CTA
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NDA/
WMA/
BLA 
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Phase I – First‐into‐Human
 20‐100 normal healthy volunteers
Determine safety and tolerability
 Determine safety and tolerability
Phase IIa/IIb – Therapeutic Exploration
 100‐300 patients
 Evaluate efficacy
 Determine dose range and adverse events (AEs)
Phase III – Therapeutic Confirmation
 1,000 to 15,000 patients
 Verify efficacy, monitor AEs and effect in usual therapy and special populations
Phase IV – Post‐Approval
 Postmarketing surveillance
 Monitor long term risks and benefits, outcomes and pharmacoeconomics
Phase V
 Postmarketing studies (Potentially thousands of patients)
 New indications: “Megatrials”
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IND/
CTA
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NDA/
WMA/
BLA 
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Phase I – First‐into‐Human
 20‐100 normal healthy volunteers
Determine safety and tolerability
 Determine safety and tolerability
Phase IIa/IIb – Therapeutic Exploration
 100‐300 patients
 Evaluate efficacy
 Determine dose range and adverse events (AEs)
Phase III – Therapeutic Confirmation
 Possibly 60‐70,000 for a vaccine
 Verify efficacy, monitor AEs and effect in usual therapy and special populations
Phase IV – Post‐Approval
 Postmarketing surveillance
 Monitor long term risks and benefits, outcomes and pharmacoeconomics
Phase V
 Postmarketing studies (Potentially thousands of patients)
 New indications: “Megatrials”
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: Rotavirus vaccine‐ 70,000 patients in 70 000 patients in
ROTATEQ® : Rotavirus vaccine
the pivotal trials worldwide
ZOSTAVAX® : Shingles vaccine‐ 68,000 patients in the pivotal trials worldwide.
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Biosimilars are to biologics what generics are to chemical
Biosimilars are to biologics what generics are to chemical
drugs
Reference
product
g
g
Biological
drug
PATENT EXPIRY
E
Chemical drug
Follow-on
version
Generic
Biosimilars ((EU))
Follow-on Biologics (FOB, US)
Subsequent-entry Biologic (SEB, Canada)
Biotechnology Follow-on Product (Japan)
Similar Biotherapeutic Product (SBP, WHO)
Follow‐on versions are generally expected to deliver price discounts.
52 | DRA NBx Training - Biosimilars | Ulrike Jägle | 28 Oct 2009 | Business Use Only
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Reduced cost of patient care – Political agenda
Business opportunity pp
y
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Expanding patient care
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Worldwide revenues for biotech drugs in 2014= $73.3
billion from sales on the top 10 approved products
Average gross margins > 75% New products –
New products 370 biotech drugs in clinical trials
370 biotech drugs in clinical trials
New markets – global expansion
Political Pressure !
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Humira (AbbVie)
$12.5
Remicade (J&J)
Remicade (J&J)
$9 2
$9.2
Rituxn (Biogen)
$8.7
Enbel (Amgen)
$8.5
Lantus (Sanofi)
$7.3
Avastin (Roche)
$7.0
Herceptin (Roche)
$6.8
Neulasta (Amgen)
l
(A
)
$5.9
$
Prevnar (Pfizer)
$4.5
Avonex (Biogen)
$3.0
$0
Source: Statista
$2
$4
$6
$8
$10
$12
$14
Revenue in Billions (US$)
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2012
2013
2014
2015
2016
2017
2018
Enbrel
Epogen/
Procrit
Remicade
Neulasta
Aranesp
Avastin
Caduet
Neupogen NovoLog
Rituxan
Humira
Pegasys
CabillyII
Avonex
Lantus
Humalog
Herceptin
Rebif
Synagis
Byetta
PegIntron
Xolair
Actemra
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To date, Europe has approved 21 biosimilars with no significant safety issues in three classes:
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Human growth hormones
Erythropoietins
Recombinant Insulin
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To date, the US FDA has received only one BLA for a biosimilar: 24 July 2014 for Neupogen (filgrastim) from Sandoz which was approved but is tied‐up in court.
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The biosimilar market in the US is not likely to see its first launch until late 2015 and will not take shape until the 2017+ timeframe. 57
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Biosimilarity – What is it?
I t h
bilit (G
i
b tit ti ?)
Interchangeability (Generic substitution?)
Patient Safety and Pharmacovigilance
Naming Convention
Use of Supportive Data and Information
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Whose data?
What is the reference product (RP) with multiple entries
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DRUG
Branded
Product
Generic #2
Generic #2
Generic #1
Branded
Product
?
BIOSIMILAR
Biosimilar #1
Biosimilar #2
?
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Biosimilarity – What is it?
Interchangeability (Generic substitution?)
Interchangeability (Generic substitution?)
Patient Safety and Pharmacovigilance
Naming Convention
Use of Supportive Data and Information
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Whose data?
What is the reference product (RP) with multiple entries
What is the reference product (RP) with multiple entries
Guidances
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EMA [2005]
FDA [2012 draft document]
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Year of Publication
Country with Specific Guidelines
2005
EU
2007
Australia
2008
Malaysia, Turkey, Taiwan
2009
Japan, Korea, Singapore, WHO
2010
Brazil, Canada, Saudi Arabia, South Africa
2011
Argentina, Cuba, India, Iran, Mexico, Peru
2012
Colombia (draft), Egypt (draft), Jordan (draft),
Thailand (draft) US (draft)
2014
EU (revised guidelines)
2015
China
Source: Boren, J., Contract Pharma 17(5): 58 (2015)
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
Requirements for extensive

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Analytical
Physico‐chemical
Biological characterization
The FDA will consider multiple factors in making study determinations:
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
Product complexity
Formulation
Product stability
Structure‐function relationships
Manufacturing process
Clinical experience with the reference product



Drugs
Synthetic, organic compounds
Defined structure, physical & chemical characteristics
chemical characteristics






Chemical synthesis
Small molecule
Stable
Homogeneous
Biotransformed (metabolized)
Activity not species specific










Acetaminophen Mw = 151
Biologics
Protein‐, carbohydrate‐ or nucleic acid ‐based product
Extracted from living organism
Complex physicochemical structure
Less well‐characterized
Macromolecule (5,000 ‐>1,000,000 )
Tertiary structure
Location, extent and type of glycosylation
Heat‐ & shear‐ sensitive
Species specific activity
Monoclonal antibody,
Mw = ~150,000
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Schellekens H. Nephrol Dial Transplant. 2005 May;20 Suppl 4:iv31-36
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•
•
•
•
The term refers to a rProtein that is in the same class as an existing biopharmaceutical but is not identical: It is i
d
th
i i l
improved over the original.
Biobetters are not entirely new drugs and they aren’t generic versions of drugs, either. Biobetters are being developed using protein or glyco‐
engineering which it is hoped will reduce the risk of immunogenicity making the drug safer, more effective and requiring a lower dose.
d
ii
l
d
A biobetter has the same target as the original but its effect on the target will hopefully last for an extended period of time.
69
•
•
•
•
Major advantage is that you are already pursuing a mechanism of action that is known and because they are considered a new drug, they will enjoy 12 years of market protection in the US unlike a biosimilar.
However, it is not clear how much payors are willing to pay for biobetters benefits.
Many believe that biosimilars will only be priced at Many believe that biosimilars will only be priced at
a 20‐25% discount off the innovator’s product.
If this is the case, one hopes the biobetters would economically rationalize a premium price.
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POTENTIAL IMPEDIMENTS TO BIOSIMILAR MARKET UPTAKE INCLUDE:

Regulatory Burden
 Name convention
 Clinical testing the FDA requires biosimilars to repeat
 Slow patient enrollment in studies

Statutory Burden
 State laws intended to restrict substitution could have the effect of hindering market uptake of biosimilars

Market Risks
Market Risks
 It is unclear how insurance companies and government health care programs will handle coverage and reimbursement
 The perception of biosimilars and the general public will play a substantial role in determining biosimilar utilization.
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“Predictions are difficult to make, especially about the future”
Yogi Berra
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dgross@padrugdiscovey.org
+1.267.893.6777
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