www.ultragenyx.com
CaSSS Gene Therapy Forum
November 8, 2018
Unique Considerations for AAV Gene Therapy Manufacturing
K. Baradaran
Legal Warning
Cautionary note regarding forward-looking statements: This presentation contains forward-looking statements, including, but
not limited to, statements regarding our plans with respect to commercializing our product and product candidates, our translational
research program, the expected timing of release of additional data for our product candidates, plans to initiate additional studies for
product candidates and timing and design of these studies, plans regarding ongoing studies for existing programs, expectations
regarding the adequacy of clinical data to support marketing applications and approvals of product candidates, our intent to file
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candidates, and expectations regarding prevalence of patients. Such forward-looking statements involve substantial risks and
uncertainties that could cause our clinical development programs, future results, performance or achievements to differ significantly
from those expressed or implied by the forward-looking statements. Such risks and uncertainties include, among others, the
uncertainties inherent in the clinical drug development process, such as the regulatory approval process, the timing of our regulatory
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estimates, and projections. Given these uncertainties, you should not place undue reliance on these forward-looking statements.
Any forward-looking statements made by us in this presentation speak only as of the date of this presentation and represent our
estimates and assumptions only as of the date of this presentation. Except as required by law, we assume no obligation, and we
disclaim any intent, to update these statements to reflect actual results.
This presentation concerns drugs that are under preclinical and/or clinical investigation and which have not yet been approved for
marketing by the U.S. Food and Drug Administration (FDA). They are currently limited by Federal law to investigational use, and no
representations are made as to their safety or effectiveness for the purposes for which they are being investigated.
Ultragenyx, Ultragenyx Pharmaceutical, Ultragenyx Gene Therapy, and our logo are our trademarks. Any other trademarks
appearing in these slides are the property of their respective holders.
1
Ultragenyx rAAV Platform for Gene Therapy
• Multiple constructs
• Oversized, Self Comp
• Codon optimization
• Enhancers
• Promoters
Transgene
Cassette
Capsids Vector Peripheral Liver
Directed
• Multiple Serotypes
• Clad E capsids Library
• Neutralizing antibodies
• Mammalian Platform
• Multiple Analytics
• Characterization
• Non-clinical (primary KO) models
• Formulations
• Internal Development & External CMO/CRO Relationships
• High Throughput Development
• Scale-Up & Tech Transfer
• Primary Liver Focus
• Global Clinical Trials & Submissions
• Expanding Indications
• Personalized Patient Delivery
Manufacturing
2
Ultragenyx rAAV Manufacturing Platforms
3
HEK293 Adherent HEK293 Suspension HeLa Producer Cell
• Well-known
• Regulatory experience
• Transient tripletransfection
• Serum-containing production
• Limited scalability (Scale-Out)
• Well-known
• Established platform
• Transient triple transfection
• Serum free production
• Bioreactors
• Scalable up to 200 to 500L, potentially scalable to 2000L
• Well-known
• Clonal, high productivity
• Similar to CHO platform
• Serum-free production
• Large scale bioreactors
• Scalable up to 2000L
▪ Three active Gene Therapy CMC/IND programs
▪ Three active global hPOC clinical trials in US and Ex-US for rare diseases
▪ Three AAV manufacturing platforms in parallel production
▪ Three INDs in less than 18 months
▪ One major collaboration
▪ Multiple CDMO
▪ Multiple CRO & Academic Labs
▪ Multiple MFG Suppliers
▪ Multiple Biorepositories
▪ Strategic CRO/CMO for clinical distribution
Topics for Discussion
▪ Typical profiles of gene therapy products vs traditional biologics
▪ Unique considerations for gene therapies vs traditional biologics
– Manufacturing
– Quality
– Supply Chain and Delivery of Product
– Comparability strategies
▪ Questions and discussion
4
Typical Profiles of Gene Therapy Products
Attribute Traditional
Biologics
Gene Therapy Cell Therapy
Disease Profile Large Rare/Small Rare/Small
Delivery Repeated Single in-vivo Single ex-vivo
Administration Multiple Primarily IV or I/T Potentially
Leukapheresis &
Infusion
Presentation Multiple Frozen Cryopreservation
Supply Chain Inventory/Multiple Personalized
Shipment/
Cold Chain
Personalized
Medicine/Cold
Chain
Production Standardized Complex A batch a patient
Clinical Trial Profile Large
Healthy
Volunteers/
Placebo/
Comparator
Small
No Healthy
Volunteers/
Open Label
Small
No Healthy
Volunteers/
Open Label
5
Commonly Used Viral Vectors*P
art
icle
Ch
ara
cte
ristics
Attribute Adenovirus AAV Herpes Retrovirus Pox/Vaccinia
Genome dsDNA ssDNA dsDNA ssRNA dsDNA
Coat Naked Naked Enveloped Enveloped Enveloped
Genome size 30-38 kb 5 kb 120-200 kb 3-9 kb 130-280 kb
Ge
ne
Th
era
py P
rop
ert
ies
Infection/
tropism
Dividing and
non-dividing
cells
Dividing
and non-
dividing
cells
Dividing
and non-
dividing
cells
Dividing
cells**
Dividing and
non-dividing
cells
Host genome
interaction
Non-
integrating
Non-
integrating
Non-
integrating
Integrating Non-integrating
Transgene expression Transient Potential
long-
lasting
Potential
long-
lasting
Long-
lasting
Transient
Packaging capacity 7.5 kb 4.5 kb >30 kb 8 kb 25 kb
Immunogenicity High High High Low High
6
*Source: White Paper on Gene Therapies 2015 Smart Analyst
** Lentivirus vectors can also infect non-dividing cells
Manufacturing Strategies Vary
▪ Modified Cell Therapy manufacturing
– Lentivirus vector
– Autologous vs Allogeneic
• CAR-T: Individual patient T-cells modified and tested before being
administered to the patient
▪ AAV manufacturing strategies
– Baculovirus
– Helper-Free Transfection
• Adherent or Suspension Triple Plasmid transient transfection
– Helper virus (HSV, AdV)
• Producer Cell Lines vs. Packaging Cell Lines
– Single vs. multiple AAV vectors (eg, zinc finger gene editing strategy)
• Multiple serotypes
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Finding the right CMO is challenging – no ‘one stop shop’
Critical Raw Materials: Traditional Biologics vs
Gene Therapy
Attribute Traditional
Biologics
Gene Therapy
MCB X X
WCB X X
Serum or Serum
Free Cell Culture
media
X X
Resins X X
Plasmids X
Viral vector(s)
stocks(X)* X
Viral Clearance X X
8
*Some viral vaccines also require virus stocks
MCB: Master Cell Bank; WCB: Working Cell Bank
MVB: Master Virus Bank; WVB: Working Virus Bank
Critical Raw Materials –
Plasmid and Helper Virus Banks
Manufacturing
Strategy
E.Coli
MCB/
WCB
E.Coli
MCB/
WCB
E.Coli
MCB/
WCB
Parent
MCB/
WCB
Product
MCB/
WCB
MVB/
WVB
Transient
transfectionx x x x
Helper virus x x x
Traditional
Biologicsx
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Need to identify ‘enabling’ CMOs for plasmid or viral vector supply
MCB: Master Cell Bank; WCB: Working Cell Bank
MVB: Master Virus Bank; WVB: Working Virus Bank
CMC Challenges & Considerations
Attribute Gene Therapy Cell Therapy
Cell & Virus Banking High High
Process VariabilityqPCR assay
dependentHigh
Batch Success (or Failure) ModerateHigh
Process is the product
Sterility Moderate Very High
Vector Transduction and
replicationHigh High
Co-packaged host DNA
High for systemic
dosing, above
WHO level (10ng
DNA/dose)
Low/Moderate
Comparability ModerateHigh
One lot per patient
Critical Raw Materials High High
10
Ultragenyx AAV Platform Supply Chain
Plasmid
•Rep
•Cap
Plasmid
•Ad Helper
Plasmid
•Trans-gene
DS
• UPS
DS
• DSP
Media
•Liquid
•Pre-made
Buffers
•Liquid
•Ready-To-Use
SUS
•Bags & Compon.
•Manifolds
•ATF
Critical RM
•Nuclease
•Resins
•Membranes
Fill Comp
•Vials
•Stoppers
•Crimpers
•Labels
Critical RM
•Nuclease
•Resins
•Membranes
DP
• Filling
• Labeling
Testing
• CDMO
• CROs
E.Coli
•MCB
•WCB
E.Coli
•MCB
•WCB
E.Coli
•MCB
•WCB
Hela S3
• MCB
• WCB
Ad5
• MVB
• WVB
HelaPCL
• MCB
• WCB
HEK293
• MCB
• WCB
Quality Testing Considerations
▪ Use of multiple QC test labs to cover wide variety of technical methods –no ‘one stop shop’
– Multiple different PCR-based methods
– Cell-based infectivity methods +/- PCR or antibody-based detection
– ELISA methods
– Gel electrophoresis +/- blotting techniques
– ICH, USP/Ph. Eur. methods
▪ Minimal sampling and sharing of vials where possible to conserve material, minimize number of QC retains
▪ Media fill results must be summarized in IND and reports provided upon request
▪ PCR method must be validated and report submitted in IND
▪ Sensitivity of release methods should be summarized in IND
▪ Characterization assays are required for each lot – and there are a lot of them
A lot of hands-on, real-time sample management and data review
Unique QC Testing Strategies:
Example Testing for 1 Lot
▪ In-process testing
– Mycoplasma
– 28-day in vitro viral assay
– Bioburden
▪ Release and characterization testing
– General (pH, appearance, osmolality)
– Identity – could be multiple methods
– Purity
– Potency – could be multiple methods
– Concentration (eg, genome copies by PCR)
– Safety testing (bioburden (DS), sterility(DP), endotoxin)
– Impurities (eg, host cell protein, host cell DNA, process residuals such as
BSA, Benzonase, affinity resin leachates)
– In addition – residual plasmid DNA, ‘packaged’ host cell DNA – multiple PCR
assays to cover different DNA sequence targets
– Sequence of final gene therapy product
13
Comparability Strategies
▪ Comparability strategies are needed for major changes, for example:
– DNA sequence modifications – coding vs non-coding regions
– Cell substrate changes (eg, HEK293 vs PCL vs Baculovirus)
– Downstream purification changes
– CMO
– Presentation or formulation changes
– Other major changes
▪ Analytical comparability typically consists of in-process, release,
characterization and stability data vs historical trends
– Side by side testing vs individual read-outs
▪ If differences are seen, non-clinical or human studies may be needed
14
Conclusions
▪ There are a number of unique CMC considerations for cell and gene therapy
manufacturing
▪ There is no ‘one stop shop’ for manufacturing and testing
▪ Different CMOs and test labs have different capabilities; capacity may be
limited
▪ Additional ‘enabling CMOs’ may be needed for critical raw materials
▪ Supply chain is complex and highly patient-centric
▪ Comparability strategies are dynamic and need to be designed to be fit for
purpose
15
Questions for Discussion
16