a quality driven biosimilar development -...
TRANSCRIPT
SEPTEMBER 17TH, 2015
A QUALITY DRIVEN BIOSIMILAR DEVELOPMENT
AN AVERSE RISK APPROACH
Amman, Jordan
September 2015 /
AFTER LUNCH TEST: FIND THE DIFFERENCE
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?
September 2015 /
CHALLENGES IN BIOSIMILAR DEVELOPMENT
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Structure
Biological Activity
Stability
Safety
Efficacy
Originator Similarity of Biosimilar
September 2015 /
OUTLINE
Understand the Complexity of Biologics
Define the Originator Quality Target Product Profile
Achieve Biosimilarity
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September 2015 /
N-GLYCOSYLATION BIOSYNTHESIS
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September 2015 /
COMPLEXITY OF N-GLYCOSYLATION IN HUMANS
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September 2015 /
TYPICAL BIOSIMILAR DEVELOPMENTS
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Reference Product
Produced in Glycans per Chain
Active Ingredient
Avastin® CHO 1xN (IgG1) Bevacizumab
Avonex® E.coli Interferon beta-1a
Benlysta® NS0 1xN (IgG1) Belimumab
Betaseron® E.coli Interferon beta-1b Copaxone chemical Glatiramer acetate
Enbrel® CHO;
3xN, 10xO (IgG1-Fus.)
Etanercept
Epogen®, Eprex® CHO; 3xN, 1xO Erythropoietin
Erbitux® SP2/0; 2xN (IgG1) Cetuximab
Gonal F® CHO; 4xN
Follicle-stimulating hormone
Herceptin® CHO; 1xN (IgG1) Trastuzumab
Humira® CHO, 1xN (IgG1) Adalimumab
Lantus® E.coli Insulin
Lucentis® E.coli Ranibizumab
Neupogen® E.coli Filgrastim
Reference Product Produced in Glycans per Chain
Active Ingredient
Nutropin® E.coli Somatropin
Octocog alpha® CHO 25xN+12xO Factor VIII
Pegasys® E.coli PEG-Interferon alpha-2a
Proleukin® E.coli Interleukin-2
Prolia® CHO 1xN (IgG2) Denosumab
Remicade® SP2/0 1xN (IgG1) Infliximab
Rituxan® CHO 1xN (IgG1) Rituximab
RoActemra® CHO 1xN (IgG1) Tocilizumab
Roferon® E.coli Interferon alpha-2a
Simponi® SP2/0 1xN (IgG1) Golimumab
Soliris® SP2/0 1xN (IgG2/4) Eculizumab
Stelara® SP2/0 1xN (IgG1) Ustekinumab
Tysabri® NS0 1xN (IgG4) Natalizumab
Xolair® CHO 1xN (IgG1) Omalizumab
Yervoy® CHO 1xN (IgG1) Ipilimumab
September 2015 /
INLFUENCE DURING MANUFACTURING
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Glycosylation
Aggregation
Deamidation
HCPs
K-Clipping
Oxidation
Fragmentation Disulfide shuffling
Glycation
Degradation
Clone development
September 2015 /
QUALITY ATTRIBUTES OF MABS
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September 2015 /
ANALYTICAL ASSAYS FOR BIOSIMILARITY ANALYSIS
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Structure/ Sequence • N- and C-terminus • Amino acid analysis • Peptide Mapping and Sequencing • Disulfide Linkage Analysis • Intact mass (reduced and middle down)
Aggregation • SEC • AUC • SDS-PAGE • AF4
Glyosylation • Monosaccharide Analysis • Oligosaccharide Analysis • Intact Mass • MALDI-TOF • HILIC-HPLC/MS
Higher Order • CD • FT-IR • NMR • H/D Exchange • X-Ray
Surface Charge • IEX • CZE • IEF/c-IEF
Activity • Bioassays • Binding assays
September 2015 /
OVERVIEW GLYCOSYLATION ANALYSIS
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Chromatographic Profiles
Saccharide Content
Glycan Pool Analysis
GlycosylationSite/ Site Occupancy/
Site Specific Pool
Glycan Structure
w/wo Deglycosylation w/wo Desialisation Degylcosylation w/wo Deglycosylation
Site specific protein fragment Exoglycosidase Defucosylation
IEF gel/cIEF Monosaccharide
Content by HPAEC-PAD
HILIC-HPLC HPAEC-PAD
Glycosylation Site and Site Occupancy by Peptide Mapping LC-ESI-MS/MS,
LC-UV
Linkage Analysis GC-MS
IEX-HPLC/RP-HPLC Sialic Acid Content by HPAEC-PAD
MALDI-MS of Released Glycans
Glycan distribution per site by Peptide Mapping
LC-ESI-MS/MS
LC-ESI/MALDI-MS/MS
2D PAGE 1D PAGE/cGE
Intact molecular weight by MALDI-MS
or LC-ESI-MS
Intact molecular weight by MALDI-MS or LC-ESI-MS NMR
September 2015 /
CHOICE TO SEE BIOSIMILARITY
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Initially fast
High risk
Some Quality Driven Development
Upfront loaded, quality driven
Lower risk
Full Quality Driven Development
Establishing Biosimilarity by scientific principles
Comparability to make a
September 2015 /
BIOSIMILAR DEVELOPEMENT - DEFINING QTPP AND CQA
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Formulation
Upstream Proc Dev
Cell Line Dev
Downstrem Proc Dev
Structural characterization e.g Amino acid sequence and compos. N/C terminus Post translational modification Glycosylation Disulfide linkage Higher order structure Aggregates
Bioassays
Physicochemical properties e.g. Molecular weight or size Isoform pattern Extinction coefficient Electrophoretic pattern Liquid chromatographic pattern Spectroscopic profiles
Ana
lytic
s
Detailed Characterization of Originator Batches
Quality Target Poduct Profile QTPP
May change during Development (e.g. Process Changes)
Critical Quality Attributes Linking Quality to Safety and Effiicacy
Enabling QbD Development Com
para
bilit
y
September 2015 /
MODULES ON REFERENCE MEDICINAL PRODUCT
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QTPP by Originator Monitoring Program
Originator AA verification
Originator Monitoring Program
Media Optimization
Upstream/ Downstream Develop
Eng. Runs and GMP Production
Pre+Clinical Phase I+III Approval Cell Line
Develop
Originator Sourcing and Logistics
Method Qualification
September 2015 /
QUALITY TARGET PRODUCT PROFILE OF 6 RMP
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Originator Batch ABC
Quality Target by Originator Batch EDF
Biosimilar Candidate
Considered not highly similar
Originator Batch ABC
Originator Batch EDF
Biosimilar Candidate
High number/high variability of characterized originator batches increases quality target range for the Biosimilar Candidate
Assumed to be highly similar
Qua
lity
Attr
ibut
e in
%
Qua
lity
Attr
ibut
e in
%
September 2015 /
CHANGE OF GLYCOSYLATION FOR MARKETED PRODUCTS
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Schiestl et al., Nature Biotechnology, Vol 29, No 4 April 2011
Rituxan/Mabthera Enbrel
September 2015 /
ONGOING ORIGINATOR MONITORING
Detailed Monitoring program of e.g. 30 RMP Lots: 10 US, 10 EU, 10 ROW
Sourced internationally with highest variability in Lots and Exp. Dates; to include different Production sites and Process Changes of RMP
Sourced and analyzed in campaigns
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September 2015 /
METHOD QUALIFICATION FOR BIOSIMILARITY
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The assays needs to be suitable to detect small differences
Generation of Stress samples
Testing of Stress samples
Final method qualification with
tested stress samples
As an example to be able to qualify selected assays a variety of stress samples will be generated and tested with two different orthogonal method.
Subsequently the samples will be used as a reference Std. for Method Qualification
September 2015 /
QTPP driven
MODULES ON BIOSIMILAR CANDIDATE
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Pool selection
Clone selection
Media Selection
Process development
Media Optimization
Upstream/ Downstream Develop
Eng. Runs and GMP Production
Pre+Clinical Phase I+III Approval Cell Line
Develop
Standardized and validated release testing
Stability testing
Comparability studies
September 2015 /
QUALITY DRIVEN DEVELOPMENT
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QTPP driven Pool selection
Clone selection
Media Selection
Process development
Initially fast
High risk
Upfront loaded, quality driven
Lower risk
Not an overall cost argument !
September 2015 /
EXERPT OF QTPP
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September 2015 /
DEVELOPMENT BY QUALITY ATTRIBUTES
Quality directed development planned in several iterations:
Clone Pool Analysis Clone Selection Media Optimization
Starting with >100 Samples and selected Quality Attributes
Screening Assays optimized for parallelization, speed and sensitivity
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Targeted attribute
Limits for this attribute
Maybe highly productive pool but not suitable
Clone Pool Single Clones Media Optimization
September 2015 /
EXAMPLE: ANALYSIS FOR CLONE SELECTION
Application: Development of an IgG1 biosimilar • MALDI-MS and subsequent comparison of glycosylation pattern • 96 samples in less than 6h (including ProteinA, Glycan Release, MALDI,
Data Evaluation)
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1605
.6
0.0
0.5
1.0
1.5
2.0
5x10
Inte
ns. [
a.u.
]
1300 1400 1500 1600 1700 1800 1900 2000 2100m/z
G0
High Mannose
G0 F
G1
G1 F
G2
G2 F
Calculated Ratios:G1/G0 and G2/G0 fucosylated/non-fucosylated
September 2015 /
NOT MEETING GLYCOSYLATION FOR BIOSIMILARITY
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-
For biosimilarity analysis, single assays can question biosimilarity.
However, demonstrating biosimilarity needs extensive characterization.
Sample E IA IB IIA IIB IIIA IIIB IV V VII VIII E
Schellekens H et al. Eur J Hosp Pharm Pract 2004;3:43–7
Eprex and Copy Biologics
September 2015 /
WHY QUALITY?
AIM: Mimimize risk starting at early development to
meet biosimilarity in the end • Saves time! • Reduces clinical trials • Mitigates later adaption of process design • Meet the authorities´ expectations
Build close collaboration between partners for optimal results
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September 2015 /
THE BIOSIMILAR GROUP CONCEPT
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June 15-18 2015 /
www.TheBiosimilarsGroup.com
The Biosimilars Group GmbH, Harrlachweg 6, 68163 Mannheim, Germany
Phone: +49 (0) 621 76449064 Email: [email protected]
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June 15-18 2015 /
Patent Situation
Development Plan
Market Access
DECISION MAKING
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