Ensuring Product Quality in Gene Transfer Clinical Trials

1. Ensuring Product Quality in Gene Transfer Clinical Trials Stephanie Simek Ph.D. Division of Cellular and Gene Therapies OTRR/CBER/FDA March 21, 2001

2. Review Research Surveillance Policy Compliance Regulation of Biological ProductsBased on Sound Science, Law and Public Health Impact

3. Regulations for Biological Products Title 21, Code of Federal Regulations • Part 312 - Investigational New Drugs (INDs) and Part 314 - New Drug Application (NDA) • Part 25 - Environmental Assessments • Part 201, 202 - Labeling & Advertising • Parts 210, 211 -Current Good Manufacturing Products (cGMPs) (FD&C Act) • Parts 610 - General Biological Product Standards (PHS Act)

4. Guidance Documents • Application of Current Statutory Authorities to Cell And Gene Therapy Products, Federal register/Vol. 58,No.1997/Oct. 14, 1993. • Guidance for Human Somatic Cell Therapy and Gene Therapy. CBER. March 1998. • PTC in the Characterization of Cell Lines Used to Produce Biologicals, 1993. 58 FR 42974. • ICH: Guidance on Viral Safety Evaluation of Biotechnology Products Derived from Cell Lines of Human or Animal Origin, FR. Sept. 24, 1998, Vol. 63, No. 185. • Guidance for Industry: Stability Testing of Drug Substances and Drug Products, (Draft Guidance), CBER, June 1998.

5. How to Ensure Product Quality • Components used in Product Manufacture • Product Testing and Characterization • Control of Manufacturing Process • cGMP Practices • In process controls

6. DEFINITION Gene Therapy: The administration of genetic material to modify or manipulate the expression of a gene product or to alter the biological properties of living cells for therapeutic use. Cells may be modified ex vivo for subsequent administration to the subject or altered in vivo by gene therapy products given directly to the subject.

7. Stages in Product Development IND Product License Phase I Phase II Phase III Phase IV Pre-IND

8. Phase I Phase II Step-wise Approach to Application of Regulatory Requirements Full characterization 21 CFR 610 ProductCharacterization Full GMP 21 CFR 210, 211 Good Manufacturing Practices Phase III Pre-clinical QA &QC, Clinical Monitoring Program Prior to Phase I : need product safety testing and basic characterization info

9. Components Used in Manufacture of Product • Vector • Cells • allogeneic & autologous cell components • Cell Bank System • master cell bank/working cell bank • master viral bank/working viral bank • Ancillary Product/Reagents • growth factors, cytokines, MoAb

10. Vector • Description, history, and detailed derivation of construct • Vector diagram • Sequence analysis

11. Cells • Autologous and allogeneic cells • Source (tissue and cell type) • Collection procedure • Donor screening • allogeneic- use blood banking criteria • autologous- don’t increase viral load or spread adventitious virus

12. Master Cell Bank • Safety Testing • Sterility • Mycoplasma • Adventitious Virus • in vitro and in vivo virus • bovine and porcine viruses • human cell lines: EBV, HBV, HCV, CMV, HIV 1&2, HTLV 1 & 2, B19, (others)

13. Master Cell Bank (cont.) • Characterization • Karyology/Morphology • Isoenzyme • Tumorgenicity • Other • Viability

14. Working Cell Bank • Safety • Sterility • Mycoplasma • In vitro Adventitious virus • Characterization • Isoenzyme • Morphology

15. Master Virus Bank • Safety Testing • Sterility • Mycoplasma • Adventitious Virus • in vitro and in vivo virus • bovine and porcine viruses • human viruses: EBV, HBV, HCV, CMV, HIV 1&2, HTLV 1 & 2, B19, (others) • murine -MAP • RCV

16. Master Virus Bank • Characterization • Identity • sequence of vector & restriction map • Activity /Expression • transgene specific protein expression • other • Titer

17. Other Reagents Used During Manufacture • Tabulation of reagents used • Final concentration • Vendor • Source (human, bovine, etc.) • Licensed product, clinical grade, reagent grade • Certificates of Analysis, cross reference letter • Qualification program

18. Product Manufacturing • Vector Production/Purification • Ex Vivo Modified Cells • method of collection/processing • ex vivo modification procedure • other modifications (irradiation) • final harvest

19. Product Manufacturing (cont.) • Formulation of Final Product • formulation buffer • excipients • vector concentration/cell density • storage

20. Final Product Testing Requirements • Demonstration of product safety • Assessment of product characterization • Maintenance of product lot consistency

21. Final Product: Safety • Sterility • Mycoplasma • Endotoxin/Pyrogenicity • Adventitious Virus • In vitro virus • RCV

22. Final Product: Characterization • Identity • restriction map, structural characterization • Activity • transgene specific • Titer • Purity • cell substrate DNA, RNA, & protein

23. Final Product Characterization (cont) • Potency • required by phase II • Stability • Development of Lot Release Specifications

24. Control of Manufacturing Process • Cell bank characterization • Master viral bank characterization • Final product characterization • Lot release tests and specifications • Ancillary products

25. Current Good Manufacturing Practices (cGMP) • Definition A set of current, scientifically sound methods, practices or principles that are implemented and documented during product development and production to ensure consistent manufacture of safe, pure and potent products • Applies to both the manufacturing process and the facilities

26. Ex Vivo Transduced CD34+ Cells Expressing HSV tk Anti-CD34+ MoAB Retroviral Viral vector Growth factors Flt-3, CSF, Fibronectin CD34+ expressing HSV tk PBSC CD34+ Selection CD34+ transduction

27. Summary • Step-wise Approach to Regulatory Requirements • Safety Testing Requirements • Control of Manufacturing Process • cGMP Practices Ensure a safe and Quality Product

28. CBER INFORMATION FAX: 301-827-3844 or 1-888-CBER-FAX PHONE: 301-827-1800 http://www.fda.gov/cber/ E-mail: CBER_INFO@A1CBER.FDA.GOVDOC-LIST@A1.CBER.FDA.GOV