Reagents Used in Cell Therapy Manufacturing Keith Wonnacott, Ph.D. Division of Cellular & Gene Therapies Office of Cellular, Tissue, and Gene Therapies

FDA Center for Biologics Evaluation and Research Center for Tobacco Products Center For Devices and Radiological Health Center for Drug Evaluation and Research National Center for Toxicological Research Office of the Commissioner Center for Biologics Evaluation and Research Center for Veterinary Medicine Center for Food Safety and Applied Nutrition Office of Chief Counsel Office of Regulatory Affairs Office of Communication, Outreach, and Development Office of Compliance and Biologics Quality Office of Management Office of Biostatistics and Epidemiology Office of Cellular, Tissue, and Gene Therapies Office of Blood Research and Review Office of Vaccines Research and Review Office of Information Technology Division of Clinical Evaluation and Pharm/Tox Division of Cell and Gene Therapies Division of Human Tissues

Terminologies Components Reagents Materials Ingredients Excipients

Components 21 CFR 210.3(b)(3) “Component means any ingredient intended for use in the manufacture of a drug product, including those that may not appear in such drug product.”

Reagents From Guidance for Human Somatic Cell Therapy INDs “Materials that are used for cell growth, differentiation, selection, purification, or other critical manufacturing steps, but are not intended to be part of the final product.” Examples: Bovine serum, Antibodies Trypsin, Collagenase, DNAse Growth factors, Cytokines Fluorescent dye Antibiotics Media/media components

Excipients From Guidance for Human Somatic Cell Therapy INDs “Any component intended to be a part of the final product” Examples: Infusion media Human serum/albumin DMSO

Other definitions raw material: Any ingredient intended for use in the production of APIs. These may include starting materials, process aids, solvents, and reagents. reagent: A substance, other than a starting material or solvent, that is used in the manufacture of an API or intermediate. active ingredient: Any component that is intended to furnish pharmacological activity or other direct effect in the diagnosis, cure, mitigation, treatment, or prevention of disease, or to effect the structure or any function of the body of man or other animals. The term includes those components that may undergo chemical change in the manufacture of the drug product and be present in the drug product in a modified form intended to furnish the specified activity or effect. Active Pharmaceutical Ingredient (API): Any substance that is represented for use in a drug and that, when used in the manufacturing, processing, or packaging of a drug, becomes an active ingredient or a finished dosage form of the drug. Such substances are intended to furnish pharmacological activity or other direct effect in the diagnosis, cure, mitigation, treatment or prevention of disease, or to affect the structure and function of the body of humans or other animals. APIs include substances manufactured by processes such as (1) chemical synthesis; (2) fermentation; (3) recombinant DNA or other biotechnology methods; (4) isolation/recovery from natural sources; or (5) any combination of these processes.

Regulatory Framework GUIDANCE REGULATION LAW

The Law

FD&C Act Section 501(a)(2)(B) “A drug or device shall be deemed to be adulterated…if it is a drug and the methods used in, or the facilities or controls used for, its manufacture, processing, packing, or holding do not conform to or are not operated or administered in conformity with current good manufacturing practice….”

Regulations

IND Regulations 21 CFR 312.23(7) “Although in each phase of the investigation sufficient information is required to be submitted to assure the proper identification, quality, purity, and strength of the investigational drug, the amount of information needed to make that assurance will vary…”

IND Regulations 21 CFR 312.23(7) “…the emphasis in an initial Phase 1 submission should generally be placed on the identification and control of the raw materials and the new drug substance…” “...As drug development proceeds…the sponsor should submit information amendments to supplement the initial information submitted…appropriate to the expanded scope of the investigation”

IND Regulations 21 CFR 312.23(7) “The submission is required to contain…a list of all components, which may include reasonable alternatives for inactive compounds, used in the manufacture of the investigational drug product, including both those components intended to appear in the drug product and those which may not appear but which are used in the manufacturing process…”

Current Good Manufacturing Practices 21 CFR 211.80 “There shall be written procedures describing in sufficient detail the receipt, identification, storage, handling, sampling, testing, and approval or rejection of components and drug product containers and closures…”

Current Good Manufacturing Practices 21 CFR 211.84 “Each lot of components…shall be withheld from use until the lot has been sampled, tested, or examined, as appropriate, and released for use by the quality control unit” “Each component shall be tested for conformity with all appropriate written specifications for purity, strength, and quality. In lieu of such testing by the manufacturer, a report of analysis may be accepted from the supplier of a component, provided that at least one specific identity test is conducted…and the reliability of the supplier’s analyses [is verified].”

Guidance

Guidance for Reviewers and Sponsors Content and Review of Chemistry, Manufacturing, and Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs) http://www.fda.gov/BiologicsBloodVaccines/GuidanceComplianceRegulatoryInformation/Guidances/Xenotransplantation/ucm074131.htm

Reagent Information for IND Tabulation of Reagents Used Qualification Program Determination of Removal of Reagents from the Final Product Other Concerns

Tabulation of Reagents Concentration of the reagent and the manufacturing step at which it is used Supplier Source Reagent Quality COA or Cross-reference letters

Reagent Qualification Define Source, Safety, and Performance “We recommend that you use FDA-approved or cleared, or clinical grade reagents whenever they are available.” “If you are using a research grade (not FDA-approved or cleared) reagent as part of the manufacturing process, we recommend that you provide information verifying the source, safety, and performance of the reagent. If the vendor of the reagent has a regulatory file with the FDA, a cross-reference letter from the sponsor may be provided in the IND. If a COA from the reagent manufacturer is used, you may assess whether the testing performed is adequate and provide that information in the IND.”

Reagent Qualification Program “If the reagent is not FDA-approved or cleared, additional testing may be needed to ensure the safety and quality of the reagent. We recommend that you establish a qualification program that includes safety testing (sterility, endotoxin, mycoplasma, and adventitious agents), functional analysis, purity testing, and assays (e.g., residual solvent testing) to demonstrate absence of potentially harmful substances. The extent of testing will depend on how the specific reagent is used in the manufacturing process.”

Specific Reagent Issues From human Albumin: ensure a licensed albumin is used and “no recalled lots were used during manufacture or preparation of the product” AB Serum: “…ensure serum is obtained from approved blood bank and meets all blood donor criteria” From recombinant cell lines Appropriate cell line testing Viral clearance when appropriate “For monoclonal antibodies, refer to the guidance on "Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use" (Ref. 9) for further information.”

Specific Reagent Issues porcine: “If porcine products are used, a COA or other documentation that the products are free of porcine parvovirus.”

Specific Reagent Issues bovine: “If a reagent is derived from bovine material, you should identify the bovine material, the source of the material, information on the location where the herd was born, raised, and slaughtered, and any other information relevant to the likelihood that the animal may have ingested animal feed prohibited under 21 CFR 589.2000. It may be that bovine material is introduced at different points in production of a reagent; the information described above should be provided for all bovine materials used.... In addition, you should provide a COA to document that bovine materials are compliant with the requirements for the ingredients of animal origin used for production of biologics described in 9 CFR 113.53.” For more information see, "Proposed Rule: Use of Materials Derived from Cattle in Medical Products Intended for Use in Humans and Drugs Intended for Use in Ruminants," (72 FR 1581; January 12, 2007) found at http://www.fda.gov/cber/rules/catruminant.htm.

Reagent Removal “You should test the final product for residual manufacturing reagents with known or potential toxicities and describe the test procedures you use to detect residual levels of these reagents in the final product. We recommend that you determine whether a qualification study is sufficient to document their removal, or whether lot release testing is appropriate prior to initiation of clinical trials.”

Guidance For Industry CGMP for Phase 1 Investigational Drugs http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm070273.pdf

QC Function “Every manufacturer should establish a written plan that describes the role of and responsibilities for QC functions. For example, a written plan should provide, at a minimum, for the following functions. Responsibility for examining the various materials used in the manufacture of a phase 1 investigational drug to ensure that they are appropriate and meet defined, relevant quality standards…”

Control of Components, and Containers and Closures “You should establish written procedures describing the handling, review, acceptance, and control of material (i.e., components, containers, closures) used in the manufacture of a phase 1 investigational drug. Materials should be controlled (e.g., segregated, labeled) until you have examined or tested the materials, as appropriate, and released them for use in manufacturing. It is important to handle and store such materials in a manner that prevents degradation or contamination.”

Control of Components, and Containers and Closures “The manufacturer should be able to identify and trace all materials used in the manufacture of a phase 1 investigational drug from receipt to use in the manufacture of each batch. We recommend that you keep a record (e.g., log book) containing relevant information on all materials. At a minimum, recorded relevant information would include receipt date, quantity of the shipment, supplier's name, material lot number, storage conditions, and corresponding expiration date.”

Control of Components, and Containers and Closures “The manufacturer should establish acceptance criteria for specified attributes on each material. For some materials, all relevant attributes or acceptance criteria may not be known at the phase 1 stage of product development. However, attributes and acceptance criteria selected for assessment should be based on scientific knowledge and experience for use in the specific phase 1 investigational drug. The material attributes and acceptance criteria will be reviewed in the IND application.”

Control of Components, and Containers and Closures “We recommend that you examine the certificate of analysis (COA) and/or other documentation on each lot of material to ensure that it meets established acceptance criteria for specified attributes. For some (e.g., human and animal derived material), documentation should include information on sourcing and/or test results for adventitious agents, as appropriate. If documentation for a material is incomplete for a specified attribute, we recommend that you test for the incomplete specified attribute of the material. For each batch of the API (or drug substance), you should perform confirmatory identity testing.”

Advice and Conclusions

Advice Know the source material and manufacturing process used to produce all reagents (particularly biological reagents) Be aware that the materials used to prepare/generate the reagents could present a potential risk Purification of protein Bacterial fermentation Ask the right questions

Conclusions Laws, regulations, and guidance help to define the appropriate use of reagents in cell therapy manufacturing The choice of reagents used as well as the qualification of those reagents in an IND is the responsibility of the sponsor Appropriate documentation of the evaluation and control of all reagents is essential for your IND

CBER Updates Final Rules Final Rule - Amendments to Sterility Test Requirements for Biological Products; Correction 5/24/2012 Final Rule - Revision of the Requirements for Constituent Materials 4/13/2011 Final Rule - Administrative Practices and Procedures; Good Guidance Practices; Technical Amendment. 4/1/2010 Expanded Access to Investigational Drugs for Treatment Use, Final Rule (Federal Register) 8/13/2009 Charging for Investigational Drugs Under an Investigational New Drug Application, Final Rule (Federal Register) 8/13/2009 Final Rule - Current Good Manufacturing Practice and Investigational New Drugs Intended for Use in Clinical Trials 7/15/2008 http://www.fda.gov/BiologicsBloodVaccines/GuidanceComplianceRegulatoryInformation/ActsRulesRegulations/General/default.htm

CBER Updates Guidances Draft Guidance of Industry: Preclinical Assessment of Investigational Cellular and Gene Therapy Products  November 2012. Guidance for Industry: Preparation of IDEs and INDs for Products Intended to Repair or Replace Knee Cartilage December 2011. Guidance for Industry: Clinical Considerations for Therapeutic Cancer Vaccines October 2011. Guidance for Industry and FDA Staff: Investigational New Drug Applications for Minimally Manipulated, Unrelated Allogeneic Placental/Umbilical Cord Blood Intended for Hematopoietic Reconstitution for Specified Indications. June 2011 Final Guidance for Industry: Potency Tests for Cellular and Gene Therapy Products January 2011. Guidance for Industry: Cellular Therapy for Cardiac Disease October 2010. Guidance for Industry - Minimally Manipulated, Unrelated Allogeneic Placental/Umbilical Cord Blood Intended for Hematopoietic Reconstitution for Specified Indications 10/2009 Guidance for Industry: Considerations for Allogeneic Pancreatic Islet Cell Products 09/2009 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

Licensed Products

(sipuleucel-T)

Fibrocell Science, Inc

HPC, Cord Blood

Product Quality - Cell Bank Apligraf is manufactured using allogeneic keratinocytes and fibroblasts derived from neonatal foreskin of human donors. Because expansion of the cell bank is limited, new banks from new donor tissue must be generated on a periodic basis. Discussion Question: Please discuss the applicant's approach to qualify and demonstrate comparability for new cell banks used for Apligraf manufacture. Product Quality – Potency For Apligraf, product potency is determined by a set of histological parameters which collectively assess the quality of the epidermal and dermal layers present in the product after maturation. These parameters include epidermal coverage, epidermal development, basal cell layer, suprabasal cell layer, dermal matrix thickness, fibroblast density and matrix aspect. These parameters were shown to correlate with percutaneous water absorption, GF/cytokine profile, cell metabolic activity by MTT, VEGF secretion and in vivo mouse assay. Discussion Question: Please discuss the use of H&E staining as a product potency measure for Apligraf.

Approval Documents http://www.fda.gov/BiologicsBloodVaccines/CellularGeneTherapyProducts/ApprovedProducts/default.htm Posted Documents Include: Package Insert Approval Letter Summary Basis for Regulatory Action Approval History, Letters, Reviews, and Related Documents

Advisory Committee Discussions Posted documents include: Agendas Briefing Documents Questions Presentations Roster Summary Transcripts Webcasts http://www.fda.gov/AdvisoryCommittees/CommitteesMeetingMaterials/BloodVaccinesandOtherBiologics/CellularTissueandGeneTherapiesAdvisoryCommittee/default.htm

Investigational Products

Total Active Investigational Cell and Gene Therapy Products (IND, IDE, MF) Fiscal Year

Yearly New IND and IDE Submissions to OCTGT

New IND and IDEs Submitted to OCTGT: Commercial or Research Sponsors

OCTGT Contact Information Cellular product manufacturing questions Keith M. Wonnacott, Ph.D. (Cell Therapies Branch Chief) keith.wonnacott@fda.hhs.gov 301-827-5102 Regulatory Questions: Contact the Regulatory Management Staff in OCTGT at CBEROCTGTRMS@fda.hhs.gov or Lori.Tull@fda.hhs.gov or by calling (301) 827-6536 OCTGT Learn Webinar Series: http://www.fda.gov/BiologicsBloodVaccines/NewsEvents/ucm232821.htm

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