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

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

I. Introduction

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, 1993. 58 FR 53248 Do not meet the criteria in 21 CFR 1271.10 to be regulated solely under PHS Act section 361 and regulations under 21 CFR 1271

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

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

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

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

II. IND Preparation

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

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

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

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

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

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 1271.90 (b)(2)) “FOR AUTOLOGOUS USE ONLY” (21 CFR Part 1271.90 (b)(1)) Processing shown not to support propagation of infectious agents

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 CFR1271.80(b)

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

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

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

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)

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

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 1271.290(b)) Appropriate patient identifiers and procedures to prevent product mix-ups Procedures in place to ensure product segregation

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

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

IV. Product Testing

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

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

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

IV. Product Testing Sterility Testing Methods Method Qualification Must be adequate to assess safety Must include aerobic, anaerobic, and fungal 610.12 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

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

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 610.9 applies) Draft Points to Consider in the Characterization of Cell Lines Used to Produce Biologicals 7/12/1993

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

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 610.13), LAL is an alternative method (21 CFR 610.9 applies)

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 600.3 (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

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?

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

V. Product Stability

V. Product Stability Stability Testing Required for IND (21 CFR 312.23(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

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

Thank you for your attention.