1. Cellular Therapy Products
Keith Wonnacott, PhD
Chief, Cellular Therapies Branch
Division of Cellular and Gene Therapies
Office of Cellular, Tissue, and Gene Therapies

2. Presentation Outline Introduction
Tips and references for IND preparation
Manufacturing information to put in an IND
Starting Material and Reagents
Product Manufacturing
Product Testing and Characterization
Product Stability
Summary and concluding remarks

3. I. Introduction

4. Introduction Somatic Cell Therapy Defined
“autologous, allogeneic, or xenogeneic cells that have been propagated, expanded, selected, pharmacologically treated, or otherwise altered in biological characteristics ex vivo to be administered to humans and applicable to the prevention, treatment, cure, diagnosis or mitigation of disease or injuries”
October 14, FR 53248
Do not meet the criteria in 21 CFR to be regulated solely under PHS Act section 361 and regulations under 21 CFR 1271

5. I. Introduction Examples of cellular therapies
Stem cells and stem cell-derived products
hematopoietic, mesenchymal, embryonic, umbilical cord blood, etc
Cancer vaccines and immunotherapies
E.g., dendritic cells, activated T lymphocytes (TIL, LAK), B lymphocytes, monocytes, cancer cells chemically modified or unmodified

6. I. Introduction More examples of cellular therapies Pancreatic islets
Chondrocytes
Keratinocytes
Hepatocytes
Xenotransplantation products
Combination Products
Gene therapy modified cells

7. I. Introduction Cell therapy opportunities OPPORTUNITIES
Cells are dynamic. They migrate, proliferate, differentiate, and respond to their environment in vitro and in vivo
Multiple cell types and mechanisms of action can be involved
Therapeutic outcome can be curative and permanent
Repair, replace, regenerate

8. I. Introduction Cell therapy challenges CHALLENGES
No terminal sterilization
Limited shelf life (due to cell viability)
Small lot size/limited sample volume
Limited availability of starting material for process, product, and test method development
Patient to patient variability and cellular heterogeneity

9. II. IND Preparation

10. II. IND Preparation Helpful hints Learn and follow the regulations
Know what’s in your product
Be data driven and include the relevant data in your submission
Present your information clearly and concisely. Tables, figures, and flow-charts can be very helpful.
Provide complete, well-organized, and internally consistent information

11. II. IND Preparation Selected regulations for cell therapy Regulation
Description
HCT/Ps
Tissue rules intended to prevent the spread of infectious disease
21 CFR 1271
Biologics
Biologics product testing standards
21 CFR 600
Drugs
IND requirements
Current good manufacturing practices
21 CFR 312
21 CFR 211

12. II. IND Preparation Examples of topics covered in guidance
CMC content and format for cell therapy
Donor Eligibility
CGMP for Phase 1
Potency
Aseptic Processing
Cell Banking
Stability
Process and test method validation

13. III. Product Manufacturing
Guidance for FDA Reviewers and Sponsors: Content and Review of Chemistry, Manufacturing, and Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs)  4/9/2008

14. III. Product Manufacturing
Cell source
Choose an appropriate cell source
Know the history, if applicable
Determine appropriate screening and testing for autologous and allogeneic donors
Minimize variability where possible

15. III. Product Manufacturing
Cell Source - Autologous
AUTOLOGOUS
Donor eligibility determination not required
However, labeling requirements may apply
“NOT EVALUATED FOR INFECTIOUS SUBSTANCES” (21 CFR Part (b)(2))
“FOR AUTOLOGOUS USE ONLY” (21 CFR Part (b)(1))
Processing shown not to support propagation of infectious agents

16. III. Product Manufacturing Cell Source - Allogeneic
Donor screening and testing required
Screened and tested per 21 CFR Part 1271
Sample timing for DE testing
Within 7 days of cell harvest/collection
Except peripheral blood stem/progenitor cells or bone marrow up to 30 days before recovery 21 CFR (b)

17. III. Product Manufacturing
Cell Source - Required Donor Testing
All
Leukocyte rich
HIV-1 (antigen and NAT)
HIV-2
HCV (antigen and NAT)
HBV (surface & core Ag)
Syphilis
HTLV-1
HTLV-2
CMV
Guidance for Industry: Eligibility Determination for Donors of Human Cells, Tissues, and Cellular and Tissue-Based Products (HCT/Ps) - 8/8/2007

18. III. Product Manufacturing Cell Source – Cell Banks
Master Cell Bank
In vivo, in vitro, and human virus testing (and potentially animal virus testing)
CMV, HIV-1 & 2, HTLV-1 & 2, EBV, HBV, HCV, B19
Bacterial, fungal, mycoplasma, endotoxin
Identity, purity, and activity testing

19. III. Product Manufacturing
Cell Source – Working Cell Bank
Working Cell Bank
In vitro virus testing
Bacterial, fungal, mycoplasma, endotoxin
Limited identity testing
ICH Guidance on Viral Safety Evaluation of Biotechnology Products Derived From Cell Lines of Human or Animal Origin - 9/24/1998

20. III. Product Manufacturing Components/Reagents/Excipients
Reagent table
Source, supplier, grade, concentration
Qualification program
Qualify reagent suitability (performs as expected)
Ensure reagent quality
Establish specifications for safety, purity, potency or reference a master file
Approve each new lot of reagents through in-house testing or review and verification manufacturer testing (COA)

21. III. Product Manufacturing
Procedures
Choose a robust manufacturing process
Establish standard operating procedures (SOPs)
Do performance runs to ensure process consistency
Establish procedures to prevent:
Product mix-ups
Product cross-contamination
Qualify procedures for safety
killing of tumorigenic cells, viral clearance, absence of replication competent virus, removal of unwanted residuals, etc.
Guidance for Industry: CGMP for Phase 1 Investigational Drugs - 7/15/2008

22. III. Product Manufacturing
Procedures - Tracking
Tracking of all products from the donor to the consignee or final disposition, and from consignee or final disposition to donor (21 CFR (b))
Appropriate patient identifiers and procedures to prevent product mix-ups
Procedures in place to ensure product segregation

23. III. Product Manufacturing
Facilities
Establish adequate facility and equipment performance standards and monitoring plans
Ensure aseptic environment for cell processing through design and monitoring
Use closed systems where possible
Use disposable equipment and process aids
Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing -- Current Good Manufacturing Practice - 9/29/2004

24. III. Product Manufacturing
Quality systems
Written procedures for ensuring manufacturing oversight
Review and approval of procedures, testing, acceptance criteria
Release/rejection of lots
Investigating manufacturing deviations
Prevent, detect, and correct deficiencies
QC recommended to be independent of production
Audit procedures and schedule

25. IV. Product Testing

26. IV. Product Testing In process testing
Provide meaningful insight into process and product quality
Contribute to the safety and quality of the final product

27. IV. Product Testing Final product testing
Needs to be performed on the final product, not intermediate
All relevant tests should be performed
Establish meaningful measures of sterility, identity, purity, and potency

28. IV. Product Testing Product characterization
Define critical product attributes
Define and monitor/control all cell types present in the product
Establish proper specifications
Ensure the safety and consistency of product lots
Base acceptance criteria on experience
Agree with current FDA standards

29. IV. Product Testing Sterility Testing Methods Method Qualification
Must be adequate to assess safety
Must include aerobic, anaerobic, and fungal
or USP <71> generally accepted
Alternative methods possible under 21 CFR 610.9
Method Qualification
Bacteriostasis and fungistasis data required if antibiotics are used in culture

30. IV. Product Testing Microbiological Testing – Rapid Release
Product may be released before 14 day culture on final product if:
In-process test (48-72 hr) is negative at time of release
Rapid method, such as Gram stain, is negative
Final product testing is initiated
Sterility failure plan is in place to ensure appropriate action and reporting

31. IV. Product Testing Mycoplasma
Testing should be done after pooling of cultures but before washing
Recommend rapid method (PCR or other) If unable to have results of culture based assay prior to administration (21 CFR applies)
Draft Points to Consider in the Characterization of Cell Lines Used to Produce Biologicals 7/12/1993

32. IV. Product Testing Identity Testing
Distinguish from other products processed in same facility
Ensure labeling is accurate
May also help to characterize the product
Distinguish between the multiple cell lines used
Define product composition through phenotypic analysis

33. IV. Product Testing Purity Testing
Freedom from extraneous material in the finished product, whether or not harmful (21 CFR 600.3(r))
Residuals contaminants
Reagents not intended to be in the product
Unwanted cellular subsets
Pyrogenicity/Endotoxin
Rabbit pyrogen method required (21 CFR ), LAL is an alternative method (21 CFR applies)

34. IV. Product Testing Potency Testing
The word potency is interpreted to mean the specific ability or capacity of the product…to effect a given result. 21 CFR (s)
A quantitative biological assay or correlated to a quantitative biological assay
Draft Guidance for Industry: Potency Tests for Cellular and Gene Therapy Products - 10/9/2008

35. IV. Product Testing Other Tests
General Safety
Cellular therapy products are exempt
Viability
Generally >70%
If not, data showing dead cells do not affect safety
Cell number/dose
Minimum, maximum?

36. IV. Product Testing Common Problems
Inappropriate timing of sample collection
Unacceptable or unqualified test method
Inadequate description of test method
Inadequate specification
Test not performed

37. V. Product Stability

38. V. Product Stability Stability Testing
Required for IND (21 CFR (a)(7)(ii))
Data generated at appropriate time and conditions
Minimally will cover time period proposed for clinical trial
Method, sampling times, temperature, assays
Should cover shipping, storage, and holding of cells at all phases of manufacturing
Stability at later phases designed to collect data to support final formulation and dating period

39. Summary Cellular therapies pose unique opportunities and challenges
CMC information should ensure quality of:
Starting Material and Reagents
Product Manufacturing
Product Testing and Characterization
Product Stability
Many additional resources are available

40. Thank you for your attention.