* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Critical Considerations for Analytical Similarity Assessment
Protein adsorption wikipedia , lookup
Biochemistry wikipedia , lookup
Multi-state modeling of biomolecules wikipedia , lookup
Protein–protein interaction wikipedia , lookup
Drug design wikipedia , lookup
Nuclear magnetic resonance spectroscopy of proteins wikipedia , lookup
List of types of proteins wikipedia , lookup
Metabolomics wikipedia , lookup
Bottromycin wikipedia , lookup
Western blot wikipedia , lookup
Intrinsically disordered proteins wikipedia , lookup
Critical Considerations for Analytical Similarity Assessment Jennifer Liu CASSS CMC Strategy Forum Japan 2015 Development of Biosimilars: Technical Aspects Biologics Are Larger and Structurally More Complex Than Chemically Synthesized Drugs Generic Same Structure Biosimilars 1.0 Similarity in Structure 2 Biosimilars 2.0 Monoclonal antibodies have multiple biological functions EU Potency Heavy chain ABP US Potency Reflective of Target Binding and Mechanism of Action Light chain Inter-chain disulfide Binding Region for FcRn (PK) N-glycan FcRn Binding Region for FcR, Effector Function (CDC, ADCC) Intra-chain disulfide CDC ADCC EU ABP US EU ABP US Biological and functional assays are used to investigate structure/functional differences which are clinically meaningful for biosimilarity Biosimilar product development begins with establishing target quality product profiles Biosimilar development is reverse engineering of the reference product Demonstrate biosimilarity in a step-wise manner Define critical quality attributes for the reference product Known mechanism of actions, biological functions, safety, and immunogenicity profiles Establish product quality profiles based on the target reference product Characterize reference product to establish targets and ranges for critical product quality attributes Develop biosimilar products to match the target reference product Match reference product profiles with greater emphasis on matching all biological functions Additional clinical data Human PK and PD Non-clinical PK/PD and toxicology as appropriate Match all predicted functions and confirm no enhancement of functions absent Demonstrate analytical similarity Comprehensive analytical similarity is critical foundation for biosmilarity Attributes related to the amino acid sequence and all post-translational modifications, including glycans Integrity of the secondary, tertiary, and quaternary structure Degradation profiles denoting stability Higher order structure Primary structure Biological and functional activities, including receptor binding and immunochemical properties Biological function Product-related substances and impurities Quantitative levels of product variants and their identities Stability General properties and excipients Properties of the finished drug product, including strength and formulation Particles and aggregates Processrelated impurities 5 Subvisible and submicron particles and aggregates of various sizes Impurities from host cells and downstream process Similarity assessment may use risk-based tier approach and statistical analysis Consider criticality ranking of quality attributes with regard to their potential impact on activity, PK/PD, and immunogenicity. Christl, L (FDA CDER) FDA Oncologic Drugs Advisory Committee meeting, 2015 Criticality Ranking Statistical Rigor High Low Tsong, Y (FDA CDER) DIA/FDA Statistics Forum 2015 6 • Pass equivalence testing if 90% confidence interval of the true mean difference is within equivalence margins • Equivalence margins = 1.50σ where σ is the true standard deviation of the RP • Statistical power (sensitivity to detect difference) dependent on number of lots Power An approach to statistical equivalence testing for tier 1 attributes Number of biosimilar batches McCamish, M. (Sandoz) Dong, X. (FDA) FDA This case study is provided for the sole purpose of illustrating FDA’s statistical approach for similarity assessment as presented at FDA ODAC meeting on Jan. 7 2015. No conclusion of the data is made by the presenter. Challenges in the tier 1 equivalence approach • Panel of Tier 1 quality attributes should be consistent for all sponsors seeking biosimilar product approval to the same original product • Relationship between quality attributes and safety/efficacy in patients may not be completely understood • Specific roles for effector function as part of the clinical mechanism of action are not entirely clear • Sufficient number of lots are required for meaningful statistical analysis • Small number of biosimilar batches reduce statistical power to detect difference • Small number of reference product lots can not estimate true variability • The true variability (standard deviation) in the reference product is difficult to estimate • Different reference product lots may come from the same drug substance lots → underestimating variability • Differences due to manufacturing process changes → overestimating variability Use appropriate characterization techniques in well-designed analytical studies is critical Reliability - Methods should be qualified and suitable for intended purposes Relevance - Methods provide meaningful information and may predict clinical performance Resolution - Methods should be sensitive and capable of resolving differences Material age - Attributes impacted by material age should be considered in the assessment Biosimilar Reference product 9 Slide from Kozlowski, S (CDER) presentation at 2014 Biomanufacturing Technology Summit, Rockville, MD, June 13, 2014 Understanding the biological relevance of measured quality attributes is important Measuring the quality attributes by retaining the important structural characteristics of the attributes Increase understanding for the biological relevance of different glycan attributes and ensure the critical attributes are controlled properly in order to match biological functions Confidence in similarity A2G0F Critical Glycans Oligosaccharide Analysis (Glycan Map) 30.0 (e.g. high-Man) A2G2F M7 A2G1F(a) A2G1F(b) M6 A1G1F 10.0 M5 A1G0F A2G0 20.0 min 0.0 Relevance 4.0 Glycan Map profile 6.0 8.0 10.0 12.0 14.0 16.0 18.0 20.0 Monosaccharide Analysis Monosaccharide analysis Reliability 10 22.0 24.0 26.0 Knowing the correlation of orthogonal methods helps develop a robust control strategy ADCC vs. Afucosylated glycan Orthogonal assays with increasing biological relevance to discern clinically meaningful differences and confirm functional similarity Confidence in manufacturing Chung, mAbs, 326-340, May 2012 NK92 ADCC High resolution and reliable methods suited for process control can guide the process and product development. Response (LU)) 20.0 10.0 0.0 4.0 Reliability 11 A2G2F M7 30.0 A2G1F(a) A2G1F(b) M6 A2G0F Glycan map M5 A1G1F Relevance FcgRIIIa binding A1G0F A2G0 PBMC ADCC min 6.0 8.0 10.0 12.0 14.0 16.0 18.0 20.0 22.0 24.0 26.0 Critical glycan attributes are modality and molecule dependent A2G0F Biosimilar epoetins: how similar are they? 30.0 Isoform 10 11 12 13 14 A2G2F M7 10.0 M5 A1G1F A2G1F(a) A2G1F(b) M6 A1G0F A2G0 20.0 min 0.0 [U/mg] RP 4.0 RP In Vivo Bioactivity 400000 350000 300000 250000 200000 150000 100000 50000 0 5 7 8 9 10 11 Isoform Number Schellekens H. Eur J Hosp Pharm Sci. 2004;3:43–47. 12 13 14 6.0 8.0 10.0 12.0 14.0 16.0 18.0 20.0 Sensitivity to difference Impact to epoetins Glycan Type Impact to mAb Sensitivity to difference High Increase clearance No glycan No ADCC Low to medium Low to medium Increase clearance Bisecting GN Increase ADCC Low to medium Medium Increase clearance High mannose and ADCC Medium High Increase clearance Terminal Gal Increase CDC Low High Decrease clearance Sialic acid Antiinflammatory and ADCC Low 12 N/A N/A Afucosylated Increase ADCC High Fingerprint-like similarity versus Fingerprint-like methods FDA Definition of Fingerprint-like: a term to describe integrated, multi-parameter approaches that are extremely sensitive in identifying analytical differences. FDA DRAFT GUIDANCE “Clinical Pharmacology Data to Support a Demonstration of Biosimilarity to a Reference Product” Koslowski, presentation at CASSS WCBP Conference, Washington DC, Jan, 31 2013 13 Analytical methods which may provide fingerprint-like profiles • Methods investigate the overall conformational integrity • • • • • 1-D NMR 2-D NMR Crystallography Antibody conformation array H/D exchange • Methods investigate the heterogeneous characteristics • Peptide map • Glycan map Can any one of these methods provide prove fingerprint-like similarity in its own right? 14 1H NMR provides fingerprint pattern • Applications for monoclonal antibodies and less complex proteins have been shown • Visual profile comparison could be subjective. An objective comparison for spectrum similarity would require complex mathematical algorithm V. Visser et. al, BioDrugs, 07 May 2013 L. Poppe, Anal. Chem. 2013, 85, 9623-9629 Similarity is assessed based on pattern similarity and little information can be discerned on the differences and their impact to purity, safety, and efficacy 15 2D-NMR has been applied for smaller nonglycosylated protein • Several publications on filgrastim (small non-glycosylated protein of ~ 19,000 Dalton) 1H-15N 2D-NMR spectral (pattern) similarity • Provides higher resolution in structural information compared to 1D-NMR down to the single amino acid level M. Levy, Anal. Bioanal. Chem. 22 Nov 2013 R. Brinson, AAPS, 2013 D. Hodgson, WCBP, 2012 Sandoz presentation to ODAC, 07 Jan. 2015 May not be suitable for glycoprotein and monoclonal antibodies which is structurally more complex than filgrastim 16 Crystallography has been applied to investigate sub-domains of large proteins • Typically require changing formulation to allow crystallization of protein of subdomains of mAb • Crystallization process generally serves as a purification step selecting the most “homogeneous” population, and could miss low abundant sub-populations S. K. Jung, mAbs 6:5, 1163--1177; September/October 2014 V. Visser et. al, BioDrugs, 07 May 2013 Does not investigate heterogeneous populations or discern sub-populations which may have different biological activities 17 H/D Exchange provides site-specific information on structural differences • H/D exchange require long experimental time with multiple testing time points. • Investigate differences in the exchange rate comparing two products under the same condition. Results could be informative in regards to detailed structural differences at specific locations Z. Zhang Anal. Chem. 2012, 84, 4942−4949 Interpretation of complex results requires experienced subject matter experts 18 Antibody conformation array could identify new epitopes due to changes in conformation • Evidences of regional conformational change may predict impact to biological activities mediated through specific antigen or receptor recognition • Reagents maybe high cost and experiment is relatively low throughput S. K. Jung, mAbs 6:5, 1163--1177; September/October 2014 X. Wang, CHI’s 6th Annual Biotherapeutics Analytical Summit Comparability for Biologics & Biosimilars March 13, 2015 Orthogonal to biological assays in investigating structure/functional differences 19 Capabilities and gaps of these fingerprint-like methods Technique Capabilities Gaps 1D-[1H]-NMR Sensitive to detect differences for both small and large proteins including monoclonal antibodies Difficult to interpret results and primarily rely on pattern similarity. May not determine specific site and levels of differences 2D-[1H-15N] HSQC NMR Sensitive to detecting differences down to amino acid levels for small proteins May not be suitable for complex biologics, such as glycoprotein and monoclonal antibodies X-ray crystallography Primary application for smaller protein or fragments of monoclonal antibody. Assess sub-populations which are homogenous in nature Limited resolution to structural differences for large protein. May omit minor components with unknown safety or efficacy impact. H/D exchange Provide detailed structural differences at specific locations Long experimental time. Interpretation of complex results requires experienced subject matter experts Ab conformational array Data analysis is less complex compared to the other fingerprinting methods. Provides regional structural information Require product-specific reagents which could be costly or may not be available These methods might provide a partial 'fingerprint' of higher order structure but are not sufficient to demonstrate 'fingerprint-like similarity’. 20 Summary • Comprehensive and well-designed similarity assessment is the foundation for biosimilarity • US FDA recommends risk-based approach for tier assignment and statistical analyses • Challenges associated with the current approach require further guidance • Biosimilar product need to demonstrate similar functions using biological assays relevant to potential clinical mechanism of action • There is no one method alone capable of indicating “fingerprint-like similarity”; it requires a collection of methods, including bioactivity assays. • “Fingerprint similarity” presumably is matching all structural/functional assessments while using comprehensive sensitive methods 21 Acknowledgements Simon Hotchin Leszek Poppe Margaret Karow Pavel Bondarenko Sundar Ramanan Zhongqi Zhang Richard Markus John Gabrielson Gino Grampp Brad Jordan Izydor Apostol Linda Narhi 22