Performance by Design: Engineering Functionality into Biopharmaceutical Products

Transcription

1 Performance by Design: Engineering Functionality into Biopharmaceutical Products Susan Dana Jones, Ph.D. Cambridge Healthtech Institute Peptalk Overcoming Expression Challenges Session San Diego, CA January 13-14, 2011

2 Enhanced Product Quality is the Goal of QbD The product is designed to meet patient needs and performance requirements The process is designed to consistently meet product critical quality attributes The impact of starting raw materials and process parameters on product quality is well understood The process is continually monitored, evaluated and updated to allow for consistent quality throughout product life cycle Critical sources of variability are identified and controlled through appropriate control strategies Ref: AS Rathore & H. Winkle, Nature Biotech 2009 H. Winkle, BPI Conference 2010

3 Traditional vs. QbD Approach Traditional Approach Quality assured by testing and inspection Data intensive submission disjointed information without big picture Specifications based on batch history Frozen process discouraging changes Focus on reproducibility often avoiding or ignoring variation QbD Approach Quality built into product & process by design, based on scientific understanding Knowledge rich submission showing product knowledge & process understanding Specifications based on product performance requirements Flexible process within design space, allowing continuous improvement Focus on robustness understanding and controlling variation

4 Design Space Determination Combine experimental data and general product knowledge to model and predict performance Use statistically designed experiments (DOEs) to determine impact of multiple parameters and their interactions Ref: C. Chen & C. Moore, FDA/Industry Statistics Workshop 2006

5 Pre-IND Phase 1: Preparing for QbD Target product profile developed in collaboration with clinical development provides essential input to process development Early process development focused on speed to clinic, not defining design space or critical process parameters Initial CQAs determined and process designed to deliver product meeting these criteria Data collection at this stage enables scale up and production for Phase 1 Design space definition is initiated at this stage Adapted from: D. Low, BPI Conference 2010

6 Can QbD be Applied to Cell Line Development? Goals of cell line development Speed: obtaining production lines quickly Titer: selecting cell lines with high titers Product Quality: limited focus on post translational modification or product heterogeneity A QbD approach could improve outcomes Product Quality: engineer cells to produce the desired posttranslational modifications and have desired properties Performance by Design QbD at CLD stage enables a rationale approach to achieving desired quality attributes

7 Availability of Clinical Material is Rate Limiting Process Development is on the critical path TASK COST MO Potency Assay Development $ 45,000 Cell Line Development $ 300,000 Cell Banking and Testing $ 164,000 Analytical Development $ 250,000 Process Development $ 600,000 Formulation Development $ 75, L Engineering Run $ 500,000 Viral Clearance Validation $ 330,000 cgmp Compliance Audits $ 40,000 Column and Resin Purchase $ 60,000 Analytical Qualification $ 150, L cgmp Manufacturing $ 900,000 Drug Product Manufacturing $ 150,000 Product Characterization $ 200,000 Stability Studies $ 300,000 Label, Package, Ship $ 150,000 Project Management $ 240,000 Non clinical studies $ 500,000 GLP Pharm/Tox $ 1,500,000 Clinical Trial Design $ 100,000 IND Preparation $ 120,000 Miscellaneous $ 1,326,000 Total Cost to IND $ 8,000,000 Submit IND to FDA Goal is to reach GLP Pharm/ Tox as quickly as possible *

8 Production Cell Line Generation is Rate Limiting Cell Line Development is on the critical path TASK COST MO Potency Assay Development $ 45,000 Cell Line Development $ 300,000 Cell Banking and Testing $ 164,000 Analytical Development $ 250,000 Process Development $ 600,000 Formulation Development $ 75, L Engineering Run $ 500,000 Viral Clearance Validation $ 330,000 cgmp Compliance Audits $ 40,000 Column and Resin Purchase $ 60,000 Analytical Qualification $ 150, L cgmp Manufacturing $ 900,000 Drug Product Manufacturing $ 150,000 Product Characterization $ 200,000 Stability Studies $ 300,000 Label, Package, Ship $ 150,000 Project Management $ 240,000 Non clinical studies $ 500,000 GLP Pharm/Tox $ 1,500,000 Clinical Trial Design $ 100,000 IND Preparation $ 120,000 Miscellaneous $ 1,326,000 Total Cost to IND $ 8,000,000 Submit IND to FDA *

9 Key features of an Initial Production Cell Line Genetic and phenotypic stability Sufficient yield to enable initiation of clinical development Yield can be improved by process development but good starting cellular productivity is essential Decision on acceptable yield should consider all requirements for material, not only clinic Desired product quality impacts selection of expression system Appropriate glycosylation or post translational processing Minimal aggregation or misfolded forms Potency

10 Defining Critical Quality Attributes (CQAs) those molecular and biological characteristics found to be useful in ensuring the safety and efficacy of the product (Q6B) Complexity of biologic products makes it difficult to define CQAs QbD focuses only on CQAs and the impact of critical process parameters on safety and efficacy Need to develop a design space to be documented in application which is based on CQAs Ref: AS Rathore & H. Winkle, Nature Biotech 2009

11 Glycosylation: When is it a CQA? Most proteins produced in mammalian cell culture are glycosylated CHO cells add human like glycoforms to N linked glycosylation sites Glycosylation may have minimal or total impact on potency, biodistribution, and safety Therapeutic antibody mechanism of action Target binding, blocking native ligand Minimal impact of glycosylation on function Activation of immune functions (ADCC, CDC) Glycosylation impacts functions and therefore efficacy

12 Overview of Antibody Structure and Function

13 Some Potential N-linked Glycoforms G0F Fucose negative G1F G1 G2F G2 Bisecting GlcNAc Acidic

14 Case Study: QbD in Cell Line Development Candidate monoclonal antibody mechanism of action primarily mediated by complement fixation More complete glycosylation (G1 and G2) correlate with increased complement fixation Product intended for chronic indication with anticipated repeat dosing Immunogenicity is a concern High mannose glycoforms are more immunogenic Target product profile based on mechanism of action and intended application Candidate cell lines generated and assessed for glycoforms during small scale bioreactor runs

15 Case Study: Experimental Approach Identify 12 lead candidate CHO cell lines with 1.5 g/l productivity Propagate cell lines in 1 L bioreactor in SFM Partial purification on Protein A DTT treatment to reduce disulfide bonds; release glycans enzymatically Analyze glycoforms using MALDI-TOF MS Select lead and back up production cell line based on glycoform analysis Candidate cell lines previously selected based on productivity and binding to target Final screen to meet TPP requirements applies QbD principles Production cell line selected based on product quality

16 Case Study: Results Clone G0F G1 G2 HighMannose 1B Low 1C High 1C Medium 4A Low 4D Very low 4D Medium 5B Medium 5B Low 7C Low-medium 8A Very low 8B Low 8B Very high

17 Designing Performance: Reduction of Fucosylation Antibodies with a high percentage of fucosylated N linked glycans can exhibit reduced ADCC activity Reduction in fucosylation during cell line development enables engineering greater ADCC performance Multiple approaches exist to reduce fucosylation POTELLIGENT Technology from Lonza uses a parental CHO cell line in which a synthetic enzyme is knocked out Advantages include documented parental line and full elimination of fucosylation GlyMaXX from ProBioGen is an alternative technology enabling fucosylation reduction in existing or new production cell lines

18 Antibody Therapeutic Activity Dechant and Valerius, Enhanced Biotherapy of Cancer, ( 2005) Volume 3, Issue 2

19 Fucose-reduced MAbs are a Natural Form Fucose AbsenceorPresenceofFucose Residue does not inflexibly alter the overall mab protein structure. No impact on PK/PD No impact on Stability No impact on Immunogenicity FUCOSE REDUCTION IS A SAFE WAY TO ENHANCE FCγRIIIA-BINDING Houde et al (Biogen Idec)

20 Fucose Synthetic Pathways and Reduction Strategy * * Deflecting Enzyme removes intermediate substrate * * Fut8

21 Pathway Deflection Depletes Fucose Efficently x CHO DG44 cells Intens. [a.u.] x Da loss Introduction of RMD gene into an established high producer clone for trastuzumab effectively removes fucose from G0, G1 and G2 structures

22 Reduced Fucosylation Increases FcγRIIIa Binding Binding assay samples G0908E EC50 [µg/ml] WT 4,33 Factor vs WT H1 0,27 16 fold H2 0,24 18 fold H3 0,22 20 fold

23 Pathway Deflection Benefits Innovative Applicable to existing or new cell lines Potent Effective minimalization with control of fucose level Universal Applicable to any mammalian cell type Healthy Cells produce same or higher titer and grow well Patent protected

24 Use of QbD in Cell Culture Development Prioritize variables for sequential DOE evaluation Data analysis simplified with limited variable set at initiation of program QbD approach to variable prioritization and definition of design space Screening: limited evaluation of 6 variables to identify critical factors Characterization: full or fractional factorial experimentation of 3 6 variables to determine inter relationship Optimization: full factorial design space determination of 2 3 critical, independent variables * Adapted from: Snee, R.D. Biopharm Intl 2010

25 Application of QbD to Cell Culture Potential Critical Process Parameters in cell culture production Temperature ph Agitation Dissolved oxygen Medium constituents Feed type and rate Multiple potential critical process parameters are evaluated in the screening stage to identify the most critical Critical process parameters are those that impact CQA

26 Challenges in Applying QbD in Early Development Time pressure provides disincentives for adding burden of process characterization early in development Availability of characterized reagents and scalable equipment may be limited in development lab Correlation of process parameters when raw material or equipment is not representative of manufacturing suite is difficult Analytical methods or operating procedures in development labs are often under developed and inappropriate to support critical process optimization decisions Defining design space with sub optimal assays is counterproductive

27 Implementing QbD in Upstream Development Instrumentation enables effective DoE to evaluate CQAs and relationship of complex variables Examples include Micro 24 and Simcell technologies Aseptic Fluidic Access Ports Compact Single-use Array Pall Micro 24 Bioreactor: Screen ml cultures Seahorse Simcell: Screen > ml cultures Six Independent MicroBioreactor Chambers Gas Permeable Transparent Windows

28 Automated SimCell Instrumentation Sampling Module Dispensing Module Sensing Module Robotic Arm Incubators

29 CQA Impact of Cell Culture Conditions Critical quality attributes of product that may change based on process parameters include: Cell viability and number Product titer Product Characteristics (e.g. glycosylation, aggregation) Impurity profile Characterization: Identify CPP through process development by evaluating impact of each parameter on the CQA Optimization: Create Design Space by optimization of critical parameters through a two factorial design of experiment

30 Optimization of Cell Culture Conditions Two factorial design monitoring product titer (yield) as a function of ph and temperature 50 conditions (10 T x 5 ph) n=9 (450 total chambers) Optimization performed using SimCell TM technology from Seahorse Corp.

31 Coordinate Upstream and Downstream Process When possible, use intended production cell line and USP conditions for initial DSP development If use non optimized bioreactor output for initial DSP work, confirm results with final USP Upstream process changes, may alter performance of downstream unit operations Change in ratio of product:contaminants Change in input ph, salt, or protein concentration Increased biomass may provide challenges in harvest or refolding

32 Phase 1 Phase 3: Risk Assessment Assess impact of process steps and conditions on final product identity, purity, potency, and safety (define CPP) Evaluate range of acceptable operational conditions for each CPP (define design space) Document product attributes from design space operations and production, including measurements of clinical performance Adapted from: D. Low, BPI Conference 2010

33 Cost and Benefit of QbD FDA Perspective From Clone to Commercial Ref: MM Nasr, ISPE National Meeting (2006)

34 Acknowledgements BioProcess Technology Consultants Sheila Magil, Ph.D. Howard Levine, Ph.D. Thomas Ransohoff ProBioGen Oliver Schub Volker Sandig, Ph.D. Hans Henning von Horsten, Ph.D.

35 Thank You! BioProcess Technology Consultants, Inc. 12 Gill Street Suite 5450 Woburn, MA USA (phone) (fax) sjones@bioprocessconsultants.com