Opportunities for Accelerating Cell Line Development and Beyond

Transcription

1 Opportunities for Accelerating Cell Line Development and Beyond European CM&C Strategy Forum May 24, 2017 Christopher Frye, Ph.D. Research Advisor & Group Leader Bioprocess R&D

2 Presentation Outline CM&C activities overview, challenges and strategies Historical CLD process and acceleration opportunities Next Generation Cell Line Development Platform (NGCLD) What is it? How can it be achieved? Potential impact of NGCLD Conclusions Acknowledgements Objective: We can significantly speed up delivery of high-quality, innovative medicines to patients! CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 2

3 CM&C Path to the Clinic is Complex Transfection Time CLD Cell Culture & Purification Banking/ Testing API Campaign DP Campaign Regulatory Production Cell Line Available MCB Thaw DP to Clinic Dev Lot CT Supply Package Q release Methods Dev & Transfer Toxicology Methods Qualification Formulation A set of interdependent activities CLD is always on critical path! CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 3

4 CM&C Strategies: Pay Me Now or Pay Me Later CM&C Strategy Advantages Disadvantages Considerations Fast-to-Data Fast-to-Launch Limit early investment speed Less late phase investment Lower comparability risk (DS & DP) Need for late phase investment Comparability risk (DS & DP) More significant early investment slower pts (clinic) Indication Timing to discharge risk Complexity of therapeutic protein CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 4

5 Commercial Cell Line at Phase 1 The production cell line is the foundation of bioprocess development directly impacting yield, product quality attributes and process robustness. Due to significant cost and risk associated with cell line switches, there is a strong business driver to develop the commercial production cell line to support Phase 1 clinical development. CLD is a complex, time-consuming, labor-intensive endeavor especially commercial production cell line development Cell line engineering capabilities can be enabling to commercial production cell line development can impact product and process-related impurity profiles impact product efficacy negatively impact development timeline CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 5

6 Historical CLD Process DNA QC/QA pclonal-derivation is >98% Electroporation Bulk Culture Selection FACS Single Cell Sorting, Growout & Consolidation ACES Screening ( 500 CDCLs) Top CDCLs in SFTS (preliminary PQA) Top 4 CDCLs (PQA) Commercial Production Cell Line 3 weeks 4 weeks 4 weeks 4 weeks weeks >25 weeks CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 6

7 Cell Line Engineering Product-Specific Challenges: Host antigen Process-Related Challenges: HCP clearance Polysorbate degradation Viral-like particles Value of Application General PQ Challenges: Sulfation Proteolysis Non-human glycosylation Available Technologies: ZFN (Zinc finger nuclease) Meganucleases TALEns RNA-based (CRISPR/Cas9, Cpf1) CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 7

8 Cell Line Engineering: Enabling Better Product Quality mab designed to bind (structural epitope) multiple human ligands Multiple hamster homologs bound Production cell line engineered (ZFN reagents) to eliminate these contaminating HCPs Phenotypically confirmed by Western blot, ELISA and LC/MS characterization Demonstrate that the genetic alterations are stable No wild-type sequence! P S P = parental cell line 1 = mono-allelic KO CDCL 2 = bi-allelic KO CDCL 3 = non-ko CDCL S = antigen standard curve (CHO) CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 8

9 Historical Cell Line Engineering Transfect (transgene) Singlecell sort Productivity screen PQ/final decision Isolate productive bulk pool Transfect editing reagent Deep-well outgrowth target screen Timeline & Resource Impact: ~3-6 months depends on starting point pts = low (<1%) high productivity bi-allelic disruption Homozygous Bi-allelic or Confirm PCR (target region) Mutation Detection Assay (MDA) NGS (NGS or Sanger) CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 9

10 Current CLD Paradigm: Acceleration Opportunities Subset of Variants chosen Candidate Selection Multiple variant start (CLD) Has minimal impact to FHD/FIH Lead Variants ID Candidate Profiling Studies MCB/ WCB GMP production FHD/FIH DNA Cell Line Development Top 4 Final Activities to FHD Bulk Pools Toxicology API production & Studies Development Lot Development Activities pooled CDCL for Toxicology API Faster to Toxicology ( fail fast ) No benefit to FHD/FIH! CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 10

11 Changing the Current Paradigm How can we move beyond these incremental tactical improvements? New technologies, capabilities and understanding have emerged over the past 3-5 years which provide substrate for NGCLD: more effective and efficient expression platform: higher productivity increased capacity reduced timeline more representative API from bulk pools You can achieve the best of both development strategies Fast-to-Data vs. Fast-to-Launch reduce timelines with no/limited resource impact or increase in technical risk What is NGCLD? CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 11

12 CLD Process: A Set of Interacting Factors High-quality Production Cell Line Parental Cell Line (PCB): Phenotypic traits Genotypic traits Time Transfection Conditions CDCL Screening: Productivity Product quality Characterization Capacity Quality Selection/Counter-Selection System(s) Clonal-Derivation Methodologies Bulk pool characterization & enrichment CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 12

13 Impact of Parental Genotypic Traits Parental cell line genotype (non-transgene) can have significant impact on cell line performance: Bulk pool format CDCLs (average of top-performing & distribution) 10-Fold More Relative Performance Relative Performance Parental 1 Parental 2 Parental 1 CDCLs Parental 2 CDCLs CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 13

14 Parental Phenotypic Traits & CLD Factor Interactions Two parental cell lines with stable, distinguishing phenotypes Evaluated in the context of two transfection conditions Consistent transfection condition-dependent performance Transfection Condition A Transfection Condition B Parent 1 > Parent 2 Parent 2 Parent 1 CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 14

15 More Effective Transfection Alternative transfection conditions: Generation of more productive (homogeneous) bulk pools Shift in CDCL distributions Transfection Condition % Bulk Pool Screen Marker % Non-producer CDCLs Transfection Condition CDCL Ave Performance Top CDCLs Ave Performance SOP 1 1 NGCLD ~3 ~3 SOP NGCLD CDCL distribution Relative Performance Relative Performance CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company NGCLD SOP 15

16 More Effective Selection Alternative selection approaches: different outcomes from different selection conditions CDCL distribution Top-producing CDCLs Selection A NGCLD SOP Selection B Selection A Selection B NGCLD SOP CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 16

17 More Efficient Screening Alternative screening approaches: elimination of traditional screen(s) significant reduction in timeline (including analytical turnaround) markers can be predictive of relative performance of cell lines enable efficient & effective identification of top cell lines very small screen sizes w/o added timeline Relative Performance Relative Performance CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 17

18 API Representativeness Improved effectiveness of NGCLD has led to highly representative API derived bulk pools relative to CDCLs Product Quality (physical & chemical attributes) % monomer, aggregate, fragments % purity, basic & acidic variants glycan profiles peptide mapping CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 18

19 Next Generation Cell Line Engineering Transfect (transgene) Deep-well outgrowth PQ/final decision Transfect editing reagent/ prime Single-cell sort/ enrich target screen Homozygous or Heterozygous Bi-allelic Timeline & Resource Impact: ~2-3 weeks vs 3-6 months pts = >10% high productivity bi-allelic disruption PCR (target region) DNA Analyzer Sanger or NGS (confirmation) CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 19

20 NGCLD Platform ACF (PCB to API)! DNA QC/QA Visual demonstration of clonal-derivation Parental Transfection & Bulk Culture Selection A B FACS enrichment, Single-cell sorting, grow out & consolidation Expansion (Screen) <24 CDCLs Productivity & PQ assessment Commercial Production Cell Line 2 weeks 3 weeks 3 weeks 7 weeks Generate engineered parental cell line CMC Strategy Forum Engineer during bulk pool selection (increased screen size) ~15 weeks Company Copyright 2017 Eli Lilly and Company 20

21 New Tactical Options Under NGCLD? NGCLD advantages: significant reduction in CLD timeline significant improvement in CLD capacity (~20-fold) Tactical options now available? commercial production cell line available for all development activities de-risk transition from Discovery>>Development limit rework # of variants Candidate Discovery Transition>>Development Tox>>>FHD Current CLD Non-CLD Development Activities Dev. Lot>Tox>>>FHD 21 NGCLD Non-CLD Development Activities CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 21

22 Challenges Will Remain NGCLD platform will impact timelines and process predictability for platform proteins (e.g. platform mabs) However challenges will remain: More complex protein scaffolds (e.g. heteromabs) potentially each having unique assembly requirements (HC: LC) will require some investment during developability stage alternatively, given increased capacity, can optimize during CLD Pressure created on host engineering earlier identification of molecule-specific issues dependent on the discovery>>>development process Creates new bottlenecks which may or may not be straightforward to eliminate CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 22

23 Conclusions New technology and process capability are enabling Next Generation CLD a more effective & efficient platform leveraging independent and synergystic improvements seamless cell line engineering capability is enabling better PQ profiles These improvements in CLD in turn provide significant value: productivity, timeline, capacity, API representativeness and manufacturing efficiencies provides options for accelerating overall clinical development timelines vision: Take CLD off the critical path to product launch Speeding delivery of high-quality, innovative medicines to patients!!!! CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 23

24 Acknowledgements Molecular & Cellular Biology Group BR&D (cross-functional) and Discovery colleagues BR&D Management Team Lonza Biologics for their licensing and permission to modify the GS Expression System Bioproduct R&D Facility, Indianapolis, Indiana CMC Strategy Forum Company Copyright 2017 Eli Lilly and Company 24