Accelerating drug development to FTIH: Potential of new expression technologies

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1 Accelerating drug development to FTIH: Potential of new expression technologies Lekan Daramola Associate Director Biopharmaceutical Development, Cell Culture & Fermentation Sciences CMC Strategy Forum Europe 2014

2 Outline Introduction Potential strategies to accelerate FTIH Can we use Transient expression platform? Pros and Cons MedImmune s proprietary CHO transient system Conclusion 2

3 Introduction Conventional method for producing biologics for IND is based on stable cell line expression platforms Cell line development is typically time consuming (6-12 months) and resource intensive Impact on accelerating drug candidates to IND Clonal stable cell lines used to produce biologic for IND for ~20 years despite significant improvements in other protein expression technologies 3

4 Stable Cell Line Based Drug Development Typical Critical Path Gene optimisation, Vector construction Transfection Extensive Screening, Bulk up Parental Cell lines Extensive Screening, Bulk up Clonal Cell lines Cell Banking 8 6 Cell bank Characterisation Phenotypic Characterisation Genetic Characterisation Product Quality Clone 1 Clone 2 Clone Clone 3 4 Choose Manufacturing cell line Bioreactor process development, Scale up Cell Bank Safety testing GMP Production CLD timeline range from 6 12 months 4

5 Potential strategies to shorten drug development timelines Synthon Non-Clonal transfected Cell lines (Parental) Stable pools (transfected cells) Plant protein expression Microbial expression Cell-free protein expression Protein Sciences - Baculovirus expression Transient expression 5

6 Transient vs. Stable Expression Systems DNA plasmid STABLE Transfection TRANSIENT Nucleus Nucleus Genomic Plasmids Integrated Plasmid Genomic Transgene DNA integrated into host genome Transfect in many copies of plasmid Expression stably maintained over many months Expression over a few days/weeks Transgene DNA passed to progeny Transcription without integration into host genome Gram-amounts but time-consuming Recombinant cell lines can be stored frozen Repeat transfection for repeat protein batches Rapid for µg to gram amounts 6

7 Why consider Transient system now? Low expression level no longer an issue Higher titres up to 2g/L reported Successful use of robust, industry standard cell line for transient expression CHO cell line Same cell host as stable cell line can now be used for Transient expression Production process transferable from stable to transient Scalable process 7

8 Expression Strategy Target Evaluation Target validation Preproject Lead Isolation Lead Optimisation Lead Profiling Pre-clinical Development Clinical Development Transient Transient gene gene expression small potential amounts >gram amounts CHO pool > gram CHO Stable cell line multi gram amounts Can Transient expression be used for GMP production?

9 Transient Expression Production Process: What it could look like? DNA GMP/non- GMP grade Cell Bulk up Transfection (Chemical or Electroporation) Grams Kg of protein Day1 Cells qualified host cell bank Transfection Production Harvest Fully characterised cells and transfected with high quality DNA Scaled up according to drug requirements Up to Day20 9

10 Benefits: Transient Material and FTIH Speed Potentially the fastest route to FTIH No cell line development required DNA purification and production phase Increase in throughput Potential increase in capacity as CLD is no longer on critical path? Organisational Reduced Cost Projects with increase rate of attrition speed Postpone resource intensive CLD to later clinical stages Potentially more cost effective to FTIH 10

11 Other Benefits of Using Transient Material Only one cell bank is required transient host cell line Same host cell line transfected with different DNA Only the host cell will require safety and genetic characterisation Wealth of information and experience from cell, gene therapy and vaccine production e.g. GMP DNA production, DNA stability, transfections Ideal for personalised medicine, low dose indications, orphan drugs Personalise d Medicine 11

12 Concerns Low expression in transient systems Process requirements: different to stable lines? Process consistency, robustness, Comparability risks Comparability with stable cell lines - later clinical stages Process development upstream and downstream Scalability Process contaminants Transfection reagent 12

13 Precedence Vaccine Production using transient transfection of HEK293 cells for clinical studies NIH FDA approved VLP production (influenza) for Phase 1 studies Working on scale up for commercial production Flu vaccine commercial production using baculovirus vector and insect cells - transient expression approach FluBlok the first FDA approved recombinant trivalent hemagglutinin vaccine. Cell/Gene therapy viral transduction, electroporation 13

14 MedImmune Proprietary CHO Transient System MedImmune has developed an industry leading proprietary CHO transient expression system Highest titres reported/published in the industry (2g/L IgG) Scalable, easy to use transfection process Successfully used to express several IgGs and non-iggs for preclinical studies over many years Improved productivity over the years as a result of continuous process development effort 14

15 Transfection Process Simple, scalable transfection protocol No media exchange required pre or post transfection High yielding process, highest to-date 2g/L of IgG i.e. potentially up to 40g crude from a 20L transfection culture Examples of >1g/L IgG titre from CHO transient platform Successfully used to fast-track research grade material production for numerous projects over the past 7 years Scaled up to 250L 15

16 Product Quality Robust process will produce consistent product quality Literature and in-house data suggest IgG product quality is comparable to material from stable cell lines Glycan data suggest transient material is comparable with stable cell line material. Within the micro-heterogeneity expected in stables e.g clone to clone micro-heterogeneity. 16

17 Conclusion Conventional stable cell generation can be time consuming Alternative expression strategies exist to accelerate timelines to FTIH Pools Cell-free expression system Different expression platforms Propose that an high yielding scalable CHO transient system is a potential viable option to accelerate timelines to FTIH Already in use for viral vaccine production e.g. viral recombinant protein, Virus-Like Particles Pros and Cons Use of Transient expression for manufacturing is now technically feasible Increase awareness 17

18 Acknowledgements Early Expression and Supply Cell Culture and Fermentation Sciences (CCFS) Group, MedImmune Ray Field Paul Varley 18