Late Phase Process Development by Applying Quality by Design

Optimising Biomanufacturing Processes, Brussels, Dec 2008 Christine Lattenmayer Group-Head Cell Biology & Media Development Biopharmaceutical Operations, Sandoz Austria

a Novartis company

Agenda
Sandoz Organization & General Novartis Concept QbD concept The 3 steps Target Definition & Redefinition Process Development Case Studies Characterization

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

A Global Network for Biopharmaceutical OperaHuningue (F) Basel (CH) Schaftenau (A) tions Clin & commercial mfg Commercial mfg Tech dev
(dedicated to Simulect DS)

Clin mfg

Vacaville (US) Clin & commercial mfg Kundl (A) Tech dev Clin & commercial mfg

Oberhaching and Holzkirchen (DE) Dev., marketing

Menges (SLO) Tech dev Commercial mfg

Mammalian cell culture Microbial fermentation

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Cell Culture CoE Schaftenau/Austria

Development of cell culture processes for new biopharmaceuticals & biosimilars Production of drug substance for clinical studies using fed-batch technology
In operation since 2004 App. 130 people
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3 USP/DSP production lines (3-13 m3) Development & analytical labs Management, quality assurance & engineering

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

The Novartis Way of Thinking: Novel biologics and biosimilars are complementary
Novel biopharmaceuticals

Novel biopharmaceuticals offer: improved treatment new therapeutic opportunities and thereby might replace older/less effective medications

Patent expiration
High quality Followon Biologics

Biosimilars offer: high quality established treatments affordable costs and thereby free up healthcare funds for new innovative drugs

Obsolescence

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

QbD concept: The 3 Steps

Define target

Development of process meeting the target, "Quality by Design"

Same concept for new biopharmaceuticals & biosimilars

Characterize result: proof that it works Product, process, clinics

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QbD concept: The 3 Steps - Same Concept for New Biopharmaceuticals & Biosimilars
New Biopharmaceutical
Product quality profile Performance Facility-fit Comparability to previous clinical DS Comparability to originator

Biosimilar

Define target

Start from standard/ platform process, rational DoE Extensive product quality analysis 2-step development (early/late phase) more time to gain experience with cell line/ product 1-step development most efficient capacity use increased challenge: rarely platform process useable, tight targets (e.g. glycosylation) to combine with high yield

Development of process

Characterize result
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Extensive set of state-of-the-art analytical methods: phys.-chem., biological characteristics Need to know structure-function relations better than originator

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

QbD concept: How to Define the Target?

Define target Target Definition & Redefinition Process Development Case Studies

Development of process meeting the target, "Quality by Design"

Characterize result: proof that it works Product, process, clinics

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Characterization

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Target definition & Redefinition: Systematic, Scientific and Risk-Based Approach

Target describes desired state of: Product quality: phys.-chem., biological, safety parameters Process performance: yield, cost-of-goods (COGS), robustness Process fitting to commercial facility Quality-target is based on risk-assessment: Define critical product quality attributes based on their expected impact on safety and efficacy Redefinition of critical quality attributes
Initial target Continuous improvement over time
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Redefined target

QbD Concept: Development of Process Meeting the Target

Define target Target Definition & Redefinition Process Development Case Studies

Development of process meeting the target, "Quality by Design"

Characterize result: proof that it works Product, process, clinics

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Characterization

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Developing a Process Meeting the Targets

Development Plan
Target, responsibilities, project plan, history, status, milestones

Focus on product quality & performance Rational DoE Interaction of parameters Product quality control

Manufacturing oriented

Facility-fit

Define target Startrun

USP media -screening DSP resin -screening

USP media-/ feed-development USP fine-tune USP parameter screening DSP parameter screening Time DSP fine-tune/ stabilities Pilotruns

Vialbreak

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Developing a Process Meeting the Targets: Rational DoE for Efficient USP Development
Approaches: Uni-variate Designs: Screening selected single parameters at very broad ranges Worst-case runs Multi-variate Designs: Full or partial factorial designs Response surface Demand-based: Replace what cells have metabolised

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Developing a Process Meeting the Targets: Rational DoE for Efficient USP Development
Investigation of main process parameters in screening and/or multivariate designs Testing of harvest stabilities at different storage conditions in screening or full-factorial designs Investigation of media components or components groups for improving titer and triggering product quality into design specifications in screening designs based on consumption rates and if appropriate, in multivariate designs Investigation of feed components or components groups for improving titer and triggering product quality into design specifications
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Case Studies
Single media-components may lead to tremendous increase of performance Increasing titer by combined medium and process optimization Adjusting charge variants by USP development Changing glycosylation pattern by innovative cell culture conditions Optimization of output of capture step Process-upscale for a sensitive mAB

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Case study 1: Single media-components may lead to tremendous increase of performance

Early Process:
commercial medium undefined composition containing peptones low product activity high process variation long main-stage (3 weeks) broad media screening single component screening (uni-variate DoE)
Productactivity [arbitrary units]
14 12 10 8

Higher Higher concentration concentration of 1 vitamin of supplement 2

Higher Higher concentration concentration of 1 trace metal of supplement 1

Final Process:
in-house medium chemically defined peptone-free high product activity short main-stage (2 weeks)
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6 4 2 0 Baseline 1st Improvement

2nd Improvement

> 10-fold increase in product activity

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Case study 2: Increasing titer by combined medium and process optimization

Phase II Process: FB with Peptone Feed
Investigation of Buffers / pH-regulation Medium supplements Feeding Osmolalities Seeding densities Temperature shift Aeration Shaking speed / Power input

Phase III Process: FB with chemically defined Feed

Titer

Pilot PhII

Manuf PhII

Osm Buffer/pH Feed

Product quality on capture eluate level SEC: PhII PhIII CEX: PhII PhIII

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Development of a biosimilar product

Define target
Refinement of target, Identification of CQA's Physicochemical and biological characterization

Development of biosimilar product, "Quality by Design"

Process development

Confirmation: Comprehensive comparability exercise

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Case Study 3: Adjusting charge variants by USP development

Charge-variants of mAb, analyzed by CEX, are typical product-related substances or impurities : acidic variants (e.g. de-amidation of Asn, cysteinylation), basic variants (e.g. amidation of Pro, Lys-variants at C-terminus) N-Terminal Cyclization (e.g. pyroglutamate at N-terminus) Adjustment via DSP is partially possible, but reduces yield Charge-variants can be adjusted via USP: media components process parameters

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Case Study 3: Adjusting charge variants by USP

Analysis of charge variants using cation exchange chromatography
pE pE pE pE pE pE pE pE Q pE pE pE pE pE pE pE Q pE pE Q

KK

CPB 2Q acidic 0K 1K 1Q 2K 2Q

0K

1K

1Q

2K

Targeting charge-variants via process parameters

Reference product range
Variant (%) Variant (%) Variant (%)

Reference product range

0 Process parameter 1
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Reference product range

0 Process parameter 2

0 Process Parameter 3

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Case Study 4: Changing glycosylation pattern by innovative cell culture conditions

Media components result in desired galactosylation state in dose-dependent manner
NP-LC of 2ABlabeled glycans

G0

G1

G2

Other media-components allow targeting of fucosylation or mannosylation

2,0

% Mannose X structure

6 5 4 3 2 1 0

Media Component B

1,5

1,0

Ref er

0,5

enc e Pr odu ct R a
0,6 0,8 1,0 1,2

0,0 0,0 0,2 0,4

nge

d7

d8

d9

d10

d7

d8

d9

d10

Concentration 1

Concentration 2

1,4

Media Component A

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Case Study 5: Optimisation of output of capture step

Improvement of washing and elution conditions of platform process conditions
Input parameter: pH of wash buffer pH of elution buffer buffer concentration of elution buffer
Estimated Response Surface buffer capacity _ elution buffer=50,0 Desirability 0,0-0,1 0,1-0,2 0,2-0,3 0,3-0,4 0,4-0,5 0,5-0,6 0,6-0,7 0,7-0,8 5,5 0,8-0,9 5,3 5,1 0,9-1,0 4,9 4,7 4,5 pH _ wash buffer

Output parameter: HCP level in eluate purity step yield elution volume

0,9 0,8 Desirability 0,7 0,6 0,5 0,4 0,3 3,2 3,3 3,4 3,5 3,6 3,7 3,8 pH _ elution buffer

choice of conditions according to multiple response optimization: pH of wash buffer pH of elution buffer buffer concentration of elution buffer

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Case study 6: Process-upscale for a sensitive mAB

The problem: CEX analysis of DS resulted in OOS
Process changes new cell line new medium different USP/DSP process 7-fold higher titer
pink: reference black: freshly thawed DS (directly frozen after production) blue: same as shown in black, 24h open (with air contact) at RT

The reason: Non-red. SDS-PAGE: reduction of Cys-bridges during storage of capture-load

Antibody reduction starts in PS.F, no full mAB is present in Capture E luate No reduced antibody was observed in DS generated during process dev. 1 2 3 4

3 - mainpeak - 19,650

1 cell-free harvest, fresh

10,0

12,5

15,0

17,5

20,0

22,5

25,0

2 cell-free harvest, stored 3 Capture eluate

4 Questions: What is root-cause for mAb-reduction? How can mAb reduction be prevented? Why was problem not apparent during process development? Slide 22 C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Drug substance

Case study 6: Process-upscale for a sensitive mAB

Root-cause for mAB reduction:
storage of cell-free harvest under N-atmosphere cellular enzymes, redox potential of spent medium, pH, sensitivity of mAb

How can mAb reduction be prevented?

storage of cell-free harvest at 4C or under airatmosphere completely prevents mAb-reduction shorten intermediate holdtime by use of larger column for capture-step

1 PS.F 0d 2 PS.F N2, 5C (2-3C), 3d 3 PS.F N2, RT, 3d

Why was problem not apparent during process development?

development was performed in air-atmosphere lesson learnt: intermediate hold-times/conditions (temperature, air/nitrogen) are simulated at small-scale for all products

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Developing a process meeting the target: Adjusting product quality of mABs

Main critical product quality attributes of mABs:
Glycoforms Charge variants Biological activity Aggregation/ Degradation

USP
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DSP

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

QbD concept: Development of process meeting the target

Define target Target Definition & Redefinition Process Development Tools & Case Studies

Development of process meeting the target, "Quality by Design"

Characterize result: proof that it works Product, process, clinics

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Characterization

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Characterize process: Proof that it works

Process characterization = Definition of design space of final process
Design-Space (ICH Q8): = multidimensional combination and interaction of input variables and process parameters that have been shown to provide a reasonable assurance of product quality; Process changes within the design-space are not considered as Regulatory Changes .

Science in combination with quality risk analyses ensure better understanding of manufacturing process and its product Sum of process parameter ranges and their interactions product quality within target specification Mainly done at small-scale with qualified small-scale model ensure

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Validate process: Proof that it works

Process validation:
Follows process characterization Large-scale process consistency runs at set-point Possibly large-scale runs at extremes of design space (e.g. holdtimes, selected parameters?) Virus clearance validation (new and used resin) Resin re-use studies

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C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

QbD concept: The 3 steps

Define target

Development of process meeting the target, "Quality by Design"

Challenging, but efficiently delivering the best process and product quality

Characterize result: proof that it works Product, process, clinics

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THANK YOU FOR YOUR ATTENTION !!!

Many thanks to
BPO Sandoz/ Schaftenau Cell Culture Center of Excellence Florian Unterluggauer Heiko Meents Jrg Windisch

Special thanks to:

Julia Schmutzhard, Corinna Sonderegger and our CMD team Josef Stettner and our BPD team Thomas Neumeier, Susanne Richter and our DSP team Wolfgang Gutleben and our analytical group
Slide 29 C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008