Integrating HOS Techniques in CQA Assessment. Not all quality attributes are critical. Abbreviations

Transcription

1 Integrating HS Techniques in CQA Assessment Tapan Das Biologics Development, Bristol-Myers Squibb April 5, th International Symposium on Higher rder Structure of Protein Therapeutics. Gaithersburg Marriott Washingtonian Center, Gaithersburg, Maryland USA 1 Not all quality attributes are critical A critical quality attribute: a physical, chemical, biological or microbiological property or characteristic that should be within an appropriate limit, range, or distribution to ensure the desired product quality - ICH Q8 (R2) ANNEX 2 Abbreviations CQA Critical Quality Attribute HS Higher rder Structure (2, 3, 4 etc) PRS Product Related Substance PRI Product Related Impurity Control strategy Control by Analytical, Process, Formulation 3 1

2 Non-CQAs often require robust control e.g. Process consistency 4 Topics Discussed Today CQA Assessment Major Quality Attributes: Assessment of Criticality Experimental Plan Integrating HS Tools in CQA Assessment Case Study Need for advanced HS Conclusions 5 A good CQA Assessment delivers built-in Quality Forced Deg, HS, Structure-Function Establish CQA Informed Decision Making Comparability Process Control, Release Assurance of Quality 6 2

3 Criticality assessment needs a focused experimental plan 7 2 Key Points RELEVANT stress studies for criticality assessment Understanding impact of one event (e.g. oxidation) at a time - on function 8 Topics Discussed Today CQA Assessment Major Quality Attributes: Assessment of Criticality Experimental Plan Integrating HS Tools in CQA Assessment Case Study Need for advanced HS Conclusions 9 3

4 Know the Molecular Profile Molecule Physicochemical properties, Isotype for mabs Structure, Homology model Identified Hot Spots (or predicted hot-spots) Molecule class knowledge MoA (Mechanism of Action) Binding, biological function any Fc-mediated/effector function any known involvement of glycans Quality Attributes & Major Degradation Events Protein Property & Degradation Aggregation %HMW Fragmentation %LMW xidation (Met, Trp) %xd Deamidation %Deam Isomerization %Iso Glycan Profile, %critical species Inherent PRS (%) thers Solution Properties Process Impurity ph Protein Conc smolality others Host Cell Protein Residual DNA others 11 Topics Discussed Today CQA Assessment Major Quality Attributes: Assessment of Criticality Experimental Plan Integrating HS Tools in CQA Assessment Case Study Need for advanced HS Conclusions 12 4

5 Experimental Plan Quality Attribute Stress Analytical HS method Methods Deamidation High ph icief, LC, PMAP CD Met xidation H22, HIL LC, PMAP CD Trp xidation AAPH, HIL PMAP CD Aggregate Various (Temp, F/T, Agitation, Enrichment etc) SEC, CE DLS, AUC Protein Particle Asp isomerization Fragment Various (Temp, F/T, Agitation, Enrichment etc) Proteindependent Various (Temp, Low ph, Enrichm) Counting, Imaging LC, PMAP SEC, CE Particle characterization, as needed CD MS 13 Acknowledgements BMS Biologics Development Hui Wei Yuetian Yan Sambit Kar Greg Barnett Gurusamy Balakrishnan Sharmistha Ghosh Ming Zeng Wei Wu HT Song Li Tao Richard Ludwig Jacob Bongers Robert Martin Mi Jin Zhengjian Li Marcel Zocher Tapan Das Reb Russell BMS Research Jano Jusuf Stanley Krystak Richard Huang Guodong Chen Adrienne Tymiak Northeastern University Roxana Iacob John Engen 14 Topics Discussed Today CQA Assessment Major Quality Attributes: Assessment of Criticality Experimental Plan Integrating HS Tools in CQA Assessment Case Study Need for advanced HS Conclusions 15 5

6 Challenge is lack of apriori knowledge which HS methods to apply 16 Large number of samples to run Selection included: CD (2 str, 3 str) DLS (large aggs) AUC (as needed) DSC (as needed) thers (HDX-MS, Raman as needed) 17 And ultimately biological function assays (an indirect proof of HS integrity) 18 6

7 Impact on HS 19 Low impact on HS High impact on HS Deamidation Isomerization Succinimide NH 2 Asn... NH NH NH... R N Succinimide... NH NH... R H 2 Iso-Asp H Asp... NH NH NH NH NH NH... R H R 20 Low impact on HS High impact on HS Terminal variants De-Glycosylation 21 7

8 Low impact on HS High impact on HS Met xidation Trp xidation 22 Low impact on HS High impact on HS Fragmentation Aggregation 23 Topics Discussed Today CQA Assessment Major Quality Attributes: Assessment of Criticality Experimental Plan Integrating HS Tools in CQA Assessment Case Study Need for advanced HS Conclusions 24 8

9 A complex case of structure-function Complex Impact of Degradation on Potency Need to apply advanced HS techniques 25 mab2 Isomerization & xidation in CDR2 Complex impact on bioactivity (Metx & IsoAsp) Aged (3+ yrs) samples were used for enrichment Isomerized mab xidized mab Native mab 26 Impact on Antigen Binding Samples %Relative Binding (to native sample) Native mab 100 Iso-mAb 13 x-mab 118 Yan et al, Analytical Chemistry, 2016, Feb 16;88(4):

10 Conformational Connectivity of Asp & Met G55 Met56 Asp54 mab 28 Asp Isomerization and Met xidation 29 Local Structure by HDX-MS 30 10

11 1 4 7 Local Conformational Changes in Region HC Native vs. isomerized native Isomerized Increased flexibility Native vs. oxidized native oxidized Decreased flexibility Global Structure by HDX-MS 32 a Global Conformational Changes Induced by D54 Isomerization (240 min) Heavy Chain Native-mAb1, HC Difference (Da) Iso-mAb1, HC -3.0 b b Light Chain Peptide number p Native-mAb1, LC 1.0 Difference (Da) Iso-mAb1, LC Peptide number 33 11

12 1 4 7 Global Conformational Changes Induced by M56 xidation (240 min) a Difference (Da) Heavy Chain Native-mAb1, HC x-mab1, HC Peptide number b Light Chain Native-mAb1, LC 1.0 Difference (Da) x-mab1, LC Peptide number pposing conformational impacts by xidation (compared to Isomerization) 34 Impact on Antigen Binding attributed to Increased Disorder in CDR by isoasp Samples %Relative Binding (to native sample) Native mab 100 Iso-mAb 13 x-mab 118 Is Asp isomerization a CQA? Establish control 35 Topics Discussed Today CQA Assessment Major Quality Attributes: Assessment of Criticality Experimental Plan Integrating HS Tools in CQA Assessment Case Study Need for advanced HS Conclusions 36 12

13 Integrating HS in CQA Assessment Selection of HS not straightforward Impact on bioactivity provides direct hint Complex structure-function may require advanced HS tools such as HDX-MS 37 THANK YU 38 13