Forced Degradation Studies for Biosimilar Development AWS
M AY 0 2 , 2 0 1 7
Forced Degradation Studies for Biosimilar Development 2017 AAPS National Biotechnology Conference, San Diego May 02, 2017
PRESENTED BY:
Amisha Kizhakkedathu, Ph.D. Associate Research Fellow Pfizer Inc. 1 Pfizer Confidential
Outline Definition of Biosimilarity FDA Approach to Demonstration of Biosimilarity Definition of Forced Degradation (FD) Scope of Forced Degradation Study (FDS)
Forced Degradation vs Formal Stability Study Scope of FD in Biosimilar Development CQA Assessment Control Strategy Method development/Validation Comparability/Similarity
Conclusion 2 Pfizer Confidential
Definition of Biosimilarity FDA Guidance 2015: Scientific Considerations in Demonstrating Biosimilarity to a Reference Product
Biosimilarity means that the biological product is highly similar to the reference product not withstanding minor differences in clinically inactive components; and
there are no clinically meaningful differences between the biological product and the reference product in terms of the safety, purity, and potency of the product
3 Pfizer Confidential
FDA Approach to Demonstration of Biosimilarity FDA Guidance 2015: Scientific Considerations in Demonstrating Biosimilarity to a Reference Product
FDA intends to consider the totality of the evidence provided by a sponsor and recommends a stepwise approach to demonstrating biosimilarity, which can include a comparison of the proposed biosimilar product and the reference product with respect to structure, function, animal toxicity, human pharmacokinetics (PK) and pharmacodynamics (PD), clinical immunogenicity, and clinical safety and effectiveness
4 Pfizer Confidential
The Goal of Biosimilar Development Is to Demonstrate That There Are No Clinically Meaningful Differences Based on the Totality of Evidence, Not to Re-establish Benefit Development Pathways Standard Biologics (Reference Product) Clinical studies
Comparative Clinical studies
Clinical pharmacology PK/PD
351(a)
Biosimilars Confirm safety profile and efficacy in a disease population (dose ranging not necessary)
Comparative clinical pharmacology PK/PD
351(k) Nonclinical Nonclinical Analytical Analytical
1. Schneider CK, et al. Nat Biotechnol. 2012;30:1179-1185. 2. McCamish M. Presented at EMA Workshop on Biosimilars; London; October 2013. 3. Berghout A. Biologicals. 2011;39:293-296. 4. US Food and Drug Adminstration. Abbreviated New Drug Applications (ANDA): Generics. http://www.fda.gov/Drugs/DevelopmentApprovalProcess/HowDrugsareDevelopedandApproved/ApprovalApplications/AbbreviatedNewDrugAppli cationANDAGenerics/. Accessed January 3, 2016. 5 Pfizer Confidential
Definition of Forced Degradation Chemical or physical stress of biopharmaceutical products to promote protein degradation
Intentional degradation of the API to an appropriate extent by means of various stress conditions such as pH, temperature, light, oxidizing agents, mechanical stress.
6 Pfizer Confidential
Scope of Forced Degradation Study (FDS) FDS provides insight into degradation pathways and degradation products of the drug that can be studied to determine the stability and critical quality attributes of the molecule Stress Condition Heat Light Oxidizing Agents Low pH High pH Freeze/Thaw Agitation …
Degradation Mechanism Fragmentation Aggregation Deamidation Isomerization Oxidation Cross-linking Cyclization Disulfide bond breakage … 7 Pfizer Confidential
Analytical Method CE-SDS SEC IEX/cIEF/CZE RP/HIC MS Peptide Mapping Potency …
Scope of Forced Degradation Study Product Degradation Pathways Stability of the Molecule CQA Assessment Impurity Characterization Developability Assessment Method Development Selection of Drug Candidate (Candidate Screening) Selection of Formulation (Formulation Development)
Selection of Process Controls (Process Development) Aids in the Implementation of Appropriate Control Strategy Selection of Stability Indicating Method
Comparability/Similarity Specification 8 Pfizer Confidential
Forced Degradation Study ≠ Stability Study Condition
Formal Stability Study
Forced Degradation
Temp: 2-8C, 25C, 40C
Temp: Typically 50C
Light: 1X ICH Q1B
Light: Varying exposure levels
Visible light 1.2M luxh + UV light 200
Wh/m2
Many other stress conditions… Study type
GMP
Non-GMP (GMP for method validation)
Subject
DS DP (Protein + Final Container/Packaging)
Drug samples from all phases of development. Protein only
Scale down models acceptable with proper matrixing/bracketing
Appropriate container (quartz vial, polypropylene tube, etc.)
Scope
Shelf-life, impact of short excursions (possible during storage/handling)
Irrelevant to shelf-life (see earlier slide on scope)
Degradant characterization
All peaks to be identified
Identification of individual degradant peaks not required
9 Pfizer Confidential
Scope of FDS in Biosimilar Development CQA Assessment Critical Quality Attributes (CQAs) are defined as “physical, chemical, biological, or microbiological properties or characteristics that should be within an appropriate limit, range, or distribution to ensure the desired product quality” [ICH Q8(R2)]. CQAs represent the attributes which are “clinically meaningful [for the] biosimilar product and the reference product in-terms of the safety, purity, and potency” [FDA Guidance]. CQAs are designed to implement appropriate control strategies to ensure manufacturing consistency, quality, safety, and efficacy of the product
CQA
Control Strategy
Product Specification Biosimilarity
Raw Materials Process Methods 10 Pfizer Confidential
Scope of FDS in Biosimilar Development CQA Assessment FDS is performed to… Reveal product degradation pathways
Stressed samples are tested in in-vitro functional assays (potency, FcRn binding etc)
NG#1 DG NG#1 NG#2
Met
NG#1
Met
Theoretical
Potential
11 Pfizer Confidential
Critical
Scope of FDS in Biosimilar Development Control Strategy Published data + In-house testing of Ref Product
Control Strategy Elements
A-MAB Case Study in Bioprocess Development, CMC Biotech Working group, 30 th October 2009 12 Pfizer Confidential
Scope of FDS in Biosimilar Development Method Development/Validation Forced Degradation Study is performed to Examine if the existing panel of analytical methods is sufficient (for the phase of the program).
Demonstrate stability indicating capability of analytical methods. Generate degradant enriched samples for method development and validation.
13 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity Comparability/Similarity Establishment Comparative Release and Stability Comparative Characterization Comparative Forced Degradation
Primary Structure, HOS Product related substances and impurities Process related impurities Drug product attributes Functional assays Other characteristics
Comparative Forced Degradation is used to Demonstrate that the degradation pathways, rates, and profiles are comparable between manufacturing processes (Comparability) between Biosimilar and Reference Product (Similarity)
Provide further evidence that the primary and higher order structure (HOS) are comparable between manufacturing processes (Comparability) between Biosimilar and Reference Product (Similarity) 14 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity
Release, Stability, Characterization
Forced Degradation
Revealing “hidden” differences
15 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Common stress conditions, regulatory expectation Heat, Light Process Driven
e.g. Agitation, Peroxide, Metal, low pH (Protein A purification) Product Driven Critical stress conditions to the product • e.g. High pH for CDR deamidation, peroxide for binding site oxidation
16 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Larger Data Set = Higher Probability of Observing Differences Lot-to-lot variability or process-to-process variability? ‒ Multiple lots per process
Concentration/formulation differences? ‒ “Bridging” samples to decouple formulation from protein concentration.
New degradation pathway or different levels of the same degradants? ‒ Characterization of isolated degradants
Consequences to safety and efficacy? ‒ Set appropriate specification for the concerned attribute 17 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Too Mild? Too Harsh? Mild
Differences: Method variability
Harsh
Appropriate stress level: Reveal “hidden” differences
Fully denatured: Similar
Coverage of mild to harsh degradation conditions in the same study • Five time points per stress, include stress control • Structure/function correlation • Kinetic plots for Comparability/Similarity (a regulatory expectation!) 18 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Forced Degradation Conditions and Time Points
Example Scenario
Type of Stress
Stress Condition
Temperature
Stress Intervals
Peroxide
0.01% H2O2 (100 ppm)
RT
0, 4, 8, 24, 32 hours
Agitation
200 rpm
RT
0, 1, 2, 4, 7 days
Light
ICH Q1B Option 2
25°C
0X, 0.2X, 0.5X, 1X, 1.5X
High pH
pH 10
RT
0, 1, 2, 4, 7 days
Freeze/Thaw
Freeze at ≤ -65 °C, Thaw at RT
19 Pfizer Confidential
0, 5 cycles
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Analytical Testing Plan Attributes
Example Scenario
Method
Peroxide
Agitation
Light
pH 10
F/T
RP-HPLC
X
X
X
X
X
SEC
-
X
X
X
X
Charge Variants
CEX
-
-
X
X
X
Size Variants
Non-reduced CE-SDS
-
X
X
X
X
Free Thiol
Free Thiol
X
-
X
X
X
Molecular Weight
Intact MS
X
X
X
X
-
Residue Specific Product Degradation
LC/MS Peptide Mapping
X
-
X
-
-
Potency
Potency assay 1
X
X
X
X
X
Potency
Potency assay 2
-
-
-
-
X
Secondary Structure
CD
-
-
-
-
X
Tertiary Structure
DSC
-
-
-
-
X
Protein Related Substances and Impurities High Molecular Weight Species
X means “Analyzed”; – means “Not Analyzed” 20 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS
Example Scenario Similarity
Comparability 20.0
Biosimilar Process-B Batch 1,2,3
% Degradant
14.0
12.0 10.0 8.0 6.0 4.0
%RRT 0.98 (RP-HPLC)
16.0
PF-06881893 PD6-267 PF-06881893 PD6-269 PF-06881893 PD6-271 Neupogen 1051319 Neupogen 1056455 Neupogen 1058392
Biosimilar Batch 1,2,3
Biosimilar Process- A Batch 1,2,3
18.0 %RRT 0.98 (RP-HPLC)
20.0
PF-06881893 2572075 PF-06881893 2575075 PF-06881893 2576075 PF-06881893 PD6-267 PF-06881893 PD6-269 PF-06881893 PD6-271
15.0
Reference Product Batch 1,2,3
10.0
5.0
2.0 0.0 0
5
10 15 20 25 Time of Peroxide Exposure (Hours)
30
0.0 35 0
21 Pfizer Confidential
5
10 15 20 25 Time of Peroxide Exposure (Hours)
30
35
Conclusion Forced Degradation (FD) is an important analytical tool in biosimilar product development, providing inputs to many filing sections. There are different types of FD studies, which serve different purposes at each stage of development.
Design FD study based on Purpose, Process, and Product. Comparative FDS is becoming an indispensable tool in demonstration of comparability/similarity.
‒ A regulatory expectation/requirement ‒ With appropriate design, can reveal “hidden” differences between products of different manufacturers and products of different processes of same manufacturer
‒ Further evidence supporting “highly similar with fingerprint-like similarity” 22 Pfizer Confidential
Questions? Amisha Kizhakkedathu, Ph.D Associate Research Fellow
Bioanalytical Development Pfizer Inc. Email: [email protected]
23 Pfizer Confidential
Acknowledgement Dr. Xiaoyu Chen
Dr. Catherine Srebalus Barnes Dr. Thomas Vanden Boom
24 Pfizer Confidential
Forced Degradation Studies for Biosimilar Development 2017 AAPS National Biotechnology Conference, San Diego May 02, 2017
PRESENTED BY:
Amisha Kizhakkedathu, Ph.D. Associate Research Fellow Pfizer Inc. 1 Pfizer Confidential
Outline Definition of Biosimilarity FDA Approach to Demonstration of Biosimilarity Definition of Forced Degradation (FD) Scope of Forced Degradation Study (FDS)
Forced Degradation vs Formal Stability Study Scope of FD in Biosimilar Development CQA Assessment Control Strategy Method development/Validation Comparability/Similarity
Conclusion 2 Pfizer Confidential
Definition of Biosimilarity FDA Guidance 2015: Scientific Considerations in Demonstrating Biosimilarity to a Reference Product
Biosimilarity means that the biological product is highly similar to the reference product not withstanding minor differences in clinically inactive components; and
there are no clinically meaningful differences between the biological product and the reference product in terms of the safety, purity, and potency of the product
3 Pfizer Confidential
FDA Approach to Demonstration of Biosimilarity FDA Guidance 2015: Scientific Considerations in Demonstrating Biosimilarity to a Reference Product
FDA intends to consider the totality of the evidence provided by a sponsor and recommends a stepwise approach to demonstrating biosimilarity, which can include a comparison of the proposed biosimilar product and the reference product with respect to structure, function, animal toxicity, human pharmacokinetics (PK) and pharmacodynamics (PD), clinical immunogenicity, and clinical safety and effectiveness
4 Pfizer Confidential
The Goal of Biosimilar Development Is to Demonstrate That There Are No Clinically Meaningful Differences Based on the Totality of Evidence, Not to Re-establish Benefit Development Pathways Standard Biologics (Reference Product) Clinical studies
Comparative Clinical studies
Clinical pharmacology PK/PD
351(a)
Biosimilars Confirm safety profile and efficacy in a disease population (dose ranging not necessary)
Comparative clinical pharmacology PK/PD
351(k) Nonclinical Nonclinical Analytical Analytical
1. Schneider CK, et al. Nat Biotechnol. 2012;30:1179-1185. 2. McCamish M. Presented at EMA Workshop on Biosimilars; London; October 2013. 3. Berghout A. Biologicals. 2011;39:293-296. 4. US Food and Drug Adminstration. Abbreviated New Drug Applications (ANDA): Generics. http://www.fda.gov/Drugs/DevelopmentApprovalProcess/HowDrugsareDevelopedandApproved/ApprovalApplications/AbbreviatedNewDrugAppli cationANDAGenerics/. Accessed January 3, 2016. 5 Pfizer Confidential
Definition of Forced Degradation Chemical or physical stress of biopharmaceutical products to promote protein degradation
Intentional degradation of the API to an appropriate extent by means of various stress conditions such as pH, temperature, light, oxidizing agents, mechanical stress.
6 Pfizer Confidential
Scope of Forced Degradation Study (FDS) FDS provides insight into degradation pathways and degradation products of the drug that can be studied to determine the stability and critical quality attributes of the molecule Stress Condition Heat Light Oxidizing Agents Low pH High pH Freeze/Thaw Agitation …
Degradation Mechanism Fragmentation Aggregation Deamidation Isomerization Oxidation Cross-linking Cyclization Disulfide bond breakage … 7 Pfizer Confidential
Analytical Method CE-SDS SEC IEX/cIEF/CZE RP/HIC MS Peptide Mapping Potency …
Scope of Forced Degradation Study Product Degradation Pathways Stability of the Molecule CQA Assessment Impurity Characterization Developability Assessment Method Development Selection of Drug Candidate (Candidate Screening) Selection of Formulation (Formulation Development)
Selection of Process Controls (Process Development) Aids in the Implementation of Appropriate Control Strategy Selection of Stability Indicating Method
Comparability/Similarity Specification 8 Pfizer Confidential
Forced Degradation Study ≠ Stability Study Condition
Formal Stability Study
Forced Degradation
Temp: 2-8C, 25C, 40C
Temp: Typically 50C
Light: 1X ICH Q1B
Light: Varying exposure levels
Visible light 1.2M luxh + UV light 200
Wh/m2
Many other stress conditions… Study type
GMP
Non-GMP (GMP for method validation)
Subject
DS DP (Protein + Final Container/Packaging)
Drug samples from all phases of development. Protein only
Scale down models acceptable with proper matrixing/bracketing
Appropriate container (quartz vial, polypropylene tube, etc.)
Scope
Shelf-life, impact of short excursions (possible during storage/handling)
Irrelevant to shelf-life (see earlier slide on scope)
Degradant characterization
All peaks to be identified
Identification of individual degradant peaks not required
9 Pfizer Confidential
Scope of FDS in Biosimilar Development CQA Assessment Critical Quality Attributes (CQAs) are defined as “physical, chemical, biological, or microbiological properties or characteristics that should be within an appropriate limit, range, or distribution to ensure the desired product quality” [ICH Q8(R2)]. CQAs represent the attributes which are “clinically meaningful [for the] biosimilar product and the reference product in-terms of the safety, purity, and potency” [FDA Guidance]. CQAs are designed to implement appropriate control strategies to ensure manufacturing consistency, quality, safety, and efficacy of the product
CQA
Control Strategy
Product Specification Biosimilarity
Raw Materials Process Methods 10 Pfizer Confidential
Scope of FDS in Biosimilar Development CQA Assessment FDS is performed to… Reveal product degradation pathways
Stressed samples are tested in in-vitro functional assays (potency, FcRn binding etc)
NG#1 DG NG#1 NG#2
Met
NG#1
Met
Theoretical
Potential
11 Pfizer Confidential
Critical
Scope of FDS in Biosimilar Development Control Strategy Published data + In-house testing of Ref Product
Control Strategy Elements
A-MAB Case Study in Bioprocess Development, CMC Biotech Working group, 30 th October 2009 12 Pfizer Confidential
Scope of FDS in Biosimilar Development Method Development/Validation Forced Degradation Study is performed to Examine if the existing panel of analytical methods is sufficient (for the phase of the program).
Demonstrate stability indicating capability of analytical methods. Generate degradant enriched samples for method development and validation.
13 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity Comparability/Similarity Establishment Comparative Release and Stability Comparative Characterization Comparative Forced Degradation
Primary Structure, HOS Product related substances and impurities Process related impurities Drug product attributes Functional assays Other characteristics
Comparative Forced Degradation is used to Demonstrate that the degradation pathways, rates, and profiles are comparable between manufacturing processes (Comparability) between Biosimilar and Reference Product (Similarity)
Provide further evidence that the primary and higher order structure (HOS) are comparable between manufacturing processes (Comparability) between Biosimilar and Reference Product (Similarity) 14 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity
Release, Stability, Characterization
Forced Degradation
Revealing “hidden” differences
15 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Common stress conditions, regulatory expectation Heat, Light Process Driven
e.g. Agitation, Peroxide, Metal, low pH (Protein A purification) Product Driven Critical stress conditions to the product • e.g. High pH for CDR deamidation, peroxide for binding site oxidation
16 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Larger Data Set = Higher Probability of Observing Differences Lot-to-lot variability or process-to-process variability? ‒ Multiple lots per process
Concentration/formulation differences? ‒ “Bridging” samples to decouple formulation from protein concentration.
New degradation pathway or different levels of the same degradants? ‒ Characterization of isolated degradants
Consequences to safety and efficacy? ‒ Set appropriate specification for the concerned attribute 17 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Too Mild? Too Harsh? Mild
Differences: Method variability
Harsh
Appropriate stress level: Reveal “hidden” differences
Fully denatured: Similar
Coverage of mild to harsh degradation conditions in the same study • Five time points per stress, include stress control • Structure/function correlation • Kinetic plots for Comparability/Similarity (a regulatory expectation!) 18 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Forced Degradation Conditions and Time Points
Example Scenario
Type of Stress
Stress Condition
Temperature
Stress Intervals
Peroxide
0.01% H2O2 (100 ppm)
RT
0, 4, 8, 24, 32 hours
Agitation
200 rpm
RT
0, 1, 2, 4, 7 days
Light
ICH Q1B Option 2
25°C
0X, 0.2X, 0.5X, 1X, 1.5X
High pH
pH 10
RT
0, 1, 2, 4, 7 days
Freeze/Thaw
Freeze at ≤ -65 °C, Thaw at RT
19 Pfizer Confidential
0, 5 cycles
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS Analytical Testing Plan Attributes
Example Scenario
Method
Peroxide
Agitation
Light
pH 10
F/T
RP-HPLC
X
X
X
X
X
SEC
-
X
X
X
X
Charge Variants
CEX
-
-
X
X
X
Size Variants
Non-reduced CE-SDS
-
X
X
X
X
Free Thiol
Free Thiol
X
-
X
X
X
Molecular Weight
Intact MS
X
X
X
X
-
Residue Specific Product Degradation
LC/MS Peptide Mapping
X
-
X
-
-
Potency
Potency assay 1
X
X
X
X
X
Potency
Potency assay 2
-
-
-
-
X
Secondary Structure
CD
-
-
-
-
X
Tertiary Structure
DSC
-
-
-
-
X
Protein Related Substances and Impurities High Molecular Weight Species
X means “Analyzed”; – means “Not Analyzed” 20 Pfizer Confidential
Scope of FDS in Biosimilar Development Comparability/Similarity: Design of FDS
Example Scenario Similarity
Comparability 20.0
Biosimilar Process-B Batch 1,2,3
% Degradant
14.0
12.0 10.0 8.0 6.0 4.0
%RRT 0.98 (RP-HPLC)
16.0
PF-06881893 PD6-267 PF-06881893 PD6-269 PF-06881893 PD6-271 Neupogen 1051319 Neupogen 1056455 Neupogen 1058392
Biosimilar Batch 1,2,3
Biosimilar Process- A Batch 1,2,3
18.0 %RRT 0.98 (RP-HPLC)
20.0
PF-06881893 2572075 PF-06881893 2575075 PF-06881893 2576075 PF-06881893 PD6-267 PF-06881893 PD6-269 PF-06881893 PD6-271
15.0
Reference Product Batch 1,2,3
10.0
5.0
2.0 0.0 0
5
10 15 20 25 Time of Peroxide Exposure (Hours)
30
0.0 35 0
21 Pfizer Confidential
5
10 15 20 25 Time of Peroxide Exposure (Hours)
30
35
Conclusion Forced Degradation (FD) is an important analytical tool in biosimilar product development, providing inputs to many filing sections. There are different types of FD studies, which serve different purposes at each stage of development.
Design FD study based on Purpose, Process, and Product. Comparative FDS is becoming an indispensable tool in demonstration of comparability/similarity.
‒ A regulatory expectation/requirement ‒ With appropriate design, can reveal “hidden” differences between products of different manufacturers and products of different processes of same manufacturer
‒ Further evidence supporting “highly similar with fingerprint-like similarity” 22 Pfizer Confidential
Questions? Amisha Kizhakkedathu, Ph.D Associate Research Fellow
Bioanalytical Development Pfizer Inc. Email: [email protected]
23 Pfizer Confidential
Acknowledgement Dr. Xiaoyu Chen
Dr. Catherine Srebalus Barnes Dr. Thomas Vanden Boom
24 Pfizer Confidential
