Outsourcing and Partnering for Commercial Grade Manufacturing Frank Tufaro, Ph.D.

Transcription

1 Outsourcing and Partnering for Commercial Grade Manufacturing Frank Tufaro, Ph.D.

2 Should You Partner Manufacturing? $$$ versus Control 2 Ph. 1 Registration Trials Commercial Launch CMO CMO or Partner CMO or Build-Out

3 Points to Consider 3 Virus family & gene insertions Should you perform process development engineering runs? Do you know likely OBD, MTD, MAD? How soon do you need this? Is virus stable in current formulation?

4 Points to Consider 4 Should you perform process development engineering runs? Do you know likely OBD, MTD, MAD? How soon do you need this? Is virus stable in current formulation?

5 Manufacturing Overview Virus Manufacturing Requires Generation of Cell and Virus Banks M A S T E R V I R US B A N K M A S T E R C E L L B A N K Extensive testing on Master Banks Confirmation of Identity Absence of adventitious agents 5 W O R K I N G V I R U S B A N K ( W V B ) W O R K I N G C E L L B A N K ( W C B ) Limited testing on Working Banks Focus on adventitious agents WVB WVB WCB WCB Purification Process Virus Lot X Virus Lot Y

6 Process Development Striking a Balance Purity Recovery Scalability Process development aims to increase purity Increased purification steps leads to decreased recovery Optimal Process Process must be fully scalable, ideally with few purification steps to facilitate process validation

7 Process Development Strategy Early phase - Non-optimized process to generate clinical supply - Quick to initiate clinical trials - Requires further development and scale-up Mid-phase - Process development and optimization - Increase scale to accommodate commercial production - Validation of Quality Control analytical methods Final phase - Validation of commercial scale process 7

8 Technical Considerations for Manufacturing Process 8 Upstream Adherent/ suspension Multi-chamber flat systems Microcarrier cultures Adapt to suspension Packed bed reactor Downstream High Capacity High Recovery Scalability Purification: adequate DNA & HCP removal Closed processing Single use Formulation Long term Stability Storage Temperature Chemistry for Compatibility & Delivery

9 Manufacturing Process Must Produce Highly Purified Virus 9 Expansion of WCB Infect with X Virus Bank Upstream Virus Production 3-6 weeks Harvest by Chemical Lysis Concentration by Ultrafiltration Digest Cellular DNA Downstream Purification 1-2 weeks Purification by Column Chromatography Sterilizing filtration; Fill in 2mL glass vials Formulation and Fill 1-2 weeks

10 Upstream Production Consistent Across Platforms 10 Similar production in cell factories, cell stacks, hyperstacks, etc. 1.00E+06 Total virus particles produced per cell Similar production at different CMOs 1.00E E+04 Upstream Production is Robust and Reproducible 1.00E+03

11 Extensive Quality Control Testing Completed on Every Lot 11 A total of 22 distinct analytical methods are employed for in-process and Quality Control (QC) release testing - Subset of QC tests completed for ongoing ICH-mandated stability testing All Release Testing is performed by audited contract laboratories All QC testing results reviewed by DNAtrix Quality Assurance (QA) Attribute Safety Purity Potency Identity QC Tests Sterility, Mycoplasma, General Safety, Bioburden, Particulate matter, in vivo adventitious virus assay, Endotoxin, adenoassociated virus assay, PCR for E1A 24bp deletion Residual Host cell DNA, Residual Host cell Protein, Residual BSA, Residual Benzonase, IEX-HPLC, SDS-PAGE Virus Particle Enumeration, Infectious titer, vp:iu ratio, Biological Activity Genomic sequence, Appearance, ph, Osmolality, SDS- PAGE, OD 260:280 ratio

12 Increased Purity Through Process Enhancements Removal of 3 major process contaminants ng/ml HCP pg/ml DNA ng/ml BSA Process 1 Process 2

13 DNX-2401 Manufacturing Efficiently Produces Purified Virus at High Yield 13 PURIFIED DNX NanoSight300 Data NS300 identifies singular particles and determines viral titer

14 TITER Stability and Formulation 14 Stability testing performed on every lot DNX-2401 stable through 36 months - testing ongoing When/If to switch formulation: lyophilize? 1.00E+12 DNX MONTH STABILITY 1.00E+11 IU/mL vp/ml 1.00E

15 Early Activities bp Replacing E3 region RGD-4C ITR E1A pcmv hox40l BGH pa Fiber ITR Clonal isolation and expansion Sequencing of virus genome OX40L expression on cell surface Bioactivity assay to examine stimulation of T cells MVB manufacturing

16 OX40L Expressed on the Surface of A549 Cells Infected with DNX Mock (PE-Isotype Control Ab) Mock (PE-hOX40L Ab) DNX-2401 (PE-hOX40L Ab) DNX-2440 (PE-hOX40L Ab) Red = Cell surface OX40L Green = Adenovirus Blue = Nucleus

17 OX40L Expressed on DNX-2440 Infected A549 Cells Triggers OX40 Stimulatory Receptor on T Cells 17 Stimulation of Jurkat/OX40 Cells by OX40L Stimulation of Jurkat/OX40 Cells by OX40L Relative Light Units Relative Light Units hr. post-infection MOI DNX-2401 DNX-2440

18 Contract Manufacturing Organizations TWO CMOs TO ENSURE SUPPLY CAN BE MET 18 Global Company; 40 facilities and 9800 Full time employees worldwide Lonza Viral Manufacturing in Houston, TX; expanding for Commercial Production SAFC (Sigma Aldrich Fine Chemicals) acquired by Merck in 2015 Global company with 50,000 Employees in 66 countries SAFC Viral Manufacturing in Carlsbad, CA; expanding for Commercial Production Potential Manufacturer in Finland

19 Biggest Challenge is Scale-Up of Virus Production to Meet Commercial Demands layer Cell Stack (CS-10) 40 CS-10

20 icellis Bioreactor Can Yield Sufficient Virus For Commercial Production Requirements icellis bioreactor designed for scale-up of adherent cell culture Surface area of icellis 500 equivalent to 3000 Roller Bottles or 800 CellStack-10s 20 = Adenovirus Lot 2 icellis 500 Surface area (m 2 ) Total virus particles produced 1.5 x x # Doses ,000

21 Should You Partner Manufacturing? Usually NO 21 CMO CMO or Partner CMO, Partner or Build-Out Ph. 1 Registration Trials Commercial Launch Better to build your own capability in late stage. Mitigating manufacturing risk is EXTREMELY important. Don t give up rights.