Accelerated Path to Probe Biology through Deferred Cloning and Applying Platform Manufacturing Processes

Transcription

1 Accelerated Path to Probe Biology through Deferred Cloning and Applying Platform Manufacturing Processes Rohini Deshpande, Ph.D., Executive Director Drug Substance Development, Amgen Thousand Oaks, CA CMC Strategy Forum Europe May 7 th, 2014

2 We need a product development strategy that meets the needs of a Tough Industry Environment Pressures on health care budgets Heightened payer focus on value Declining R&D productivity Global regulatory expectations Lengthy timelines for product development High cost to develop drugs due to low success rate

3 Objectives of Accelerated Process Development Reach go/no go decisions early by probing biology in humans Enhance first-to-market probability Increase pipeline capacity Generate representative material efficiently Allow phase appropriate PD and Reg CMC investment

4 Legacy Process: Opportunity for Phase Appropriate Approach Project Strategy Team Molecule Assessment FIH process development Clinical Production Phase I/II studies Commercial Process Development Cell Bank 1 and Process 1 Cell Bank 1 and Process 2 IND Clinical Production INDa Pivotal studies Dual cycle investment for drug substance process development (P1 for FIH to P2 for commercial) Single cycle approach for cell line development (commercial ready cell lines at FIH opportunity?) Can we shorten cycle time by moving to a dual cycle approach for cell line development?

5 Generating phase appropriate clinical material earlier: Dual Cycle Cell Line Development Transfections, selection and amplification Cell Pools Cell bank 1 Toxicology and early phase clinical studies Single cell cloning Deferred Cloning Cell bank 2 Final clone selection Pivotal studies and commercialization

6 E B B A B B Process Dev. Platform and High Throughput Tools Compose and test A C A C A C A C D Database In silico designs Platform verification/set point definition & supply Candidate selection to FIH: platform process for representative material Cell line and process selection Purification process definition

7 Phase appropriate approach saves time for Tox and FIH material supply Accelerated approach Molecule, methods, process, and formulation Lead Cell bank 1 and DS manufacturing Traditional approach Lead Molecule selection Methods, process, and formulation Cell bank1 and DS manufacturing Deferred Cloning and platform application reduces time to IND by ~8 months For Internal Use Only. Amgen Confidential. 7

8 Deferred Cloning & Comparability Accelerated approach PST IND INDa Molecule, process, formulation, and methods Tox and Clinical Prod Process 1 and Cell Bank 1 Material generated from cell pools Phase I/II studies CPD Process 2 and Cell Bank 2 Material generated from clonal cell lines Clinical Prod Pivotal studies Is the product quality data for material generated from cell pools and clones comparable? Does it pose a risk to comparability? For Internal Use Only. Amgen Confidential. 8

9 Cell bank 1 from cell pools: Measures for fit-for-use Plasmid quality is controlled and sequencing performed on single isolates Multiple transfections are performed to ensure many parental cell pool options Product quality attributes are tested using sensitive LC/MS/MS methods to detect microheterogeneity and confirm primary sequence Product consistency is achieved by using defined population doublings for each run (one vial thaw/run) ICH guidelines are followed for cell bank genetic characterization (DNA sequencing), release testing, and adventitious agent testing For Internal Use Only. Amgen Confidential. 9

10 Cell Bank 1 created from a cell pool will follow the same testing paradigm (no change is proposed) Purpose Tests CB1 Identity Isoenzyme analysis X Safety Sterility X Genetic stability Mycoplasma In vitro viral assay In vivo viral assay Transmission electron microscopy of cells Mouse antibody production Hamster antibody production Cocultivation with reverse transcriptase and focus endpoints for retrovirus detection In vitro assay for bovine viruses* In vitro assay for porcine viruses* cdna sequence confirmation X X X X X X X X X X**

11 Expression system is designed to generate stable cell pools & clones Antibody and selection marker are produced as a single mrna Only cells with functional selection marker survive Provides a tight linkage between antibody light and heavy chain genes and the survival gene i.e. they exist as part of the same mrna Leads to the creation of highly productive and homogeneous stable pools which can be used for largescale production

12 Count Frequency Count Frequency Cell Pool Productivity & Homogeneity Amgen Expression System Alternate Expression System LC HC+LC HC Non-expressing population Mock pool Cell pool 5 4 Expressing Cells APC-A Antibody Secretion* *Flow cytometry Assay APC-A Antibody Secretion* GMD_30min.fcs

13 Similar FACS profile between cell pool and clones Grey- Pool Red- Clone 1 Blue- Clone 2 Antibody Secretion Homogeneity of Cell Pools is an Enabler for the Accelerated Approach

14 (g/l) Efficient generation of tox and GMP supply Titer GITR BAFF IL2-Mutein Cell pools 800 VCD Culture Duration (Days) High titers from cell pools allows use of a single lot for tox and FIH supply

15 Modality independent platform analytical technology Proteins Total Ion Chromatogram Disulfide integrity Charge variant Digest Clipping Extracted Ion Chromatograms Identity Glycans Chemical modification

16 Analytical methodologies allow detection of sequence variants from cell pools Multi-attribute method Change from profile to attributes Utilize Orbitrap MS Detect low level protein microheterogeneity Quantify desired attributes Detect Unknowns Fully automated Suitable for product disposition Replaces conventional methods Fewer DS/DP release/stability testing No sequence variants were detected in 3 pilot programs from material produced using cell pools

17 % HMW % Galactosylation (A2G1F) % a-fucosylated % High Mannose Cell pools & clones: similar PQA ranges Cell Pools Clones Cell Pools Clones Cell Pools Clones Cell Pools Clones Over 20 cell pools are generated for diversity and over hundred clones are screened to ensure comparability

18 Molecule 1: Comparable PQAs for material from cell pools & clones Purity Assay Attribute Cell pool Clone Size Exclusion Ionexchange rce-sds FIH Spec (DS/DP) HMW < 5% Acidic Main ± 10% Basic LC+HC % LMW N/A NGHC N/A Glycan Afucosylation High Mannose N/A Impurity CHOP ng/ml < 200 ppm DNA pg/mg N/A < ng/dose Activity Bioassay Potency % *Difference in CEX acid and main is from change in harvest operation

19 AU Molecule 1: Comparable Product Quality for material from cell pools & clones Cell pool clone Minutes Peaks aligned for visualization No significant new peaks or peak changes. Slight differences in missed cleavage, minor PTMs, and glycosylation

20 Molecule 2: Comparable PQAs for material from cell pools & clones Assay Attribute Amplified pool (3-150) Clone*1 Clone*2 FIH Spec (DS/DP) Size Exclusion HMW < 5% Acidic Ion-exchange Main ± 10% Basic LC+HC % rce-sds LMW N/A Purity NGHC N/A Pre-Peaks < 5% nrce-sds %HH N/A %HHL N/A % Afucosylation Glycan % High Mannose N/A % Galactosylation Impurity CHOP ng/mg 50 < 200 ppm DNA pg/mg 0.7 < 10 ng/dose Potency Binding % %

21 Molecule 3: Comparable PQAs for material from cell pools & clones Assay Attribute Cell pool Clone 1 Clone 2 FIH Spec (DS/DP) Size Exclusion HMW < 5% Ionexchange Acidic Main Basic ± 10% Purity rce-sds LC+HC % Pre-Peaks < 5% nrce-sds %Main N/A Glycan % Afucosylation 1.6 % High Mannose 6.4 % Galactosylation N/A

22 Molecule 3: similar PQA profile from vial thaw to end of production bioreactors for cell pool and clones Pool 6 Clone HMW A-Fucosylation Galactosylation High Mannose VCD Titer 0 PreMCB MCB Start of Production Bioreactor End of Production Bioreactor

23 In Summary, Product Quality from Cell Pools is comparable to clones Both cell sources performed similarly: Cell Culture Process Purification Process Product Quality Attributes Purity /Impurity Stability Manufacturing and Quality controls are used to ensure no/low risk to patient safety and product consistency Next steps Engagement and feedback from the regulatory agencies Application to molecules amenable to platform Molecules with less complex product quality attributes Risk based approached to comparability

24 Biosimilar Development Transcends the Highest Risk Category of Process Change; experience can be applied to advancing NMEs Innovator Process Change Governed by comparability guidance (ICH Q5E), Para 2.2 of CHMP 1 Nature of Process Change Change filter Raw material supplier Replace equipment Site Change Change cell culture media Cell line or formulation change Biosimilar development requires most if not all changes Risk & Data Requirements Abbreviated comparability Low Risk Commonly implemented - Analytical data - Process studies Comprehensive comparability Moderate Risk High Risk Less commonly implemented - Analytical data - Analytical data - Process studies - Process studies - Stability data - Stability data - Clinical data Biosimilars Out of scope of comparability guidance (ICH Q5E) Covered by EMA, FDA, and WHO Biosimilar Guidelines [Adapted from presentation by G. Grampp (Amgen); Manufacturing Changes in the Era of Biosimilars, BioProcess International Conference, November 2, 2011, Long Beach, CA] Biosimilars Different: Facility and equipment Cell-line, vector and culture media Fermentation/culture conditions Downstream processing/purification Formulation and container/closure

25 Acknowledgements Discovery Research Translational Sciences Clinical Operations & Operations Technology Regulatory Affairs (CMC) Product Quality

26 Questions