Comparability and Setting Product Specification Limits – A Case Study Bo Kara, Process Development Advanced Therapy Delivery UKRMP- Comparability: Manufacturing, Characterisation and Controls Workshop, Cambridge, UK. 14-15 September, 2015

Outline

1. GSK and Cell Gene Therapy 2. Control Strategy 3. Case Study: • ADA-SCID Gene Therapy Manufacturing Process and Changes • Comparability Study Design • Comparability Study Results and Conclusions

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GSK - Cell and Gene Therapy Platform Alliance (2010) GSK- TIGET • Retroviral ex-vivo gene therapy for ADA SCID (lead - optioned 2010) – Adenosine deaminase (ADA) deficiency, Severe Combined Immunodeficiency (SCID)

• Lentiviral platform for lysosomal storage disorders, primary immune deficiencies and blood disorders: 2 optioned 4Q2013 (MLD, WAS) – Metachromatic leukodystrophy (MLD), Wiskott-Aldrich Syndrome (WAS)

• Telethon responsible for advancing all programs to PoC • GSK responsible for global Regulatory, Manufacture and Commercial activity • Driven by GSK’s commitment to R+D and diversification

Collaboration with Adaptimmune (2014) • TCR engineered T cell immunotherapies for cancer

(The San Raffaele Telethon Institute for Gene Therapy (TIGET), joint-venture between TIGET and the Telethon Foundation)

“Control Strategy” - Underpins the Product Specification “The commercial manufacturing process(es) are controlled by ensuring process parameters (including critical process parameters), in-process controls and in process specifications are within their expected ranges” •

Process Parameter – an independent variable that may be controlled during the execution of an experiment or manufacturing process

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Critical Process Parameters (CPP’s) – parameters whose variability may have an impact on a Critical Quality Attribute (CQA) and therefore should be monitored or controlled to ensure the process produces a product of the desired quality

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In-Process Controls (IPC’s) – tests/activities performed during production to monitor, and if necessary and appropriate, adjust, the manufacturing process

Deviations form pre-defined parameters or control ranges will be assessed to determine their impact on the relevant CQA(s) prior to batch deposition •

In-Process Specifications – tests performed during production to ensure the product will be of the desired quality

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Product Specifications – (chosen) to confirm the quality of the DS and DP rather than to establish full characterisation: • molecular and biological characteristics demonstrated to support ensuring the safety and efficacy of the product

Deviations from the in-process/product specifications will trigger and Out of Specification (OOS) investigation

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Demonstrating Comparability of Clinical and Commercial Process – Case Study ADA-SCID Autologous Hematopoietic Stem Progenitor Cell (HSPC) Gene Therapy

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Manufacturing Logistics Vector Manufacturing Cell Processing/Manufacturing site (MolMed s.p.a.) colocated with Hospital/Treatment site (San Raffaele Hospital, Milan, Italy)

BM Harvest Joint Accreditation Committee - International Society for Cellular Therapy and European Society for Blood and Marrow Transplantation (JACIE). Patients tested : presence of infectious agents as EU Directives 2004/23/EC and 2006/17/EC

Sterile filtered vector aliquots stored at 80% • Clinical and commercial process batches comparable • Results within historical range

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Comparability Results Cell Process - ADA Activity • Healthy donor BM with background ADA activity • Results subtracted from background observed in non-transduced samples • Clinical batches lower than commercial batches (lower VCN)

✓

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Comparability Results Cell Process – Additional Characterisation • Provirus sequence confirmation • Integration site analysis by LAM-PCR (linear amplification mediated-PCR): polyclonal appearance for both clinical and commercial batches • Extensive phenotype characterisation using multiple cell surface differentiation markers - CD45, CD34, CD90, CD133, CD19, CD16/CD56, CD15, CD3

✓

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Comparability Results Cell Process - Residuals and Impurities • Acceptance criteria set as Commercial ≤ Clinical for all residuals • Residual BSA, Retronectin, cytokines - comparable levels for all batches - Greater clearance from DS to DP for commercial batches due to the wash step • HCP, Host Cell-DNA, free vector particles (total and infectious) - Higher in commercial DS batches (due to higher viral titres used for transduction), but reduced to