FDA

1. FDA�s Office of Critical Path Programs (OCPP) The Clinical Trials Transformation Initiative (CTTI) FDA�s Clinical Investigator Course

3. Drug Substance & Product [312.23(a)(7)] A description of the drug substance, including its� characteristics A list of all components�used in the manufacture� Drug substance and drug product manufacturer information The general method of preparation� The acceptable limits and analytical methods used to assure the identity, strength, quality, and purity� [specifications] Information to support the stability of the drug� during the � proposed study(ies)

4. 1? structure higher order structure post-translational modifications heterogeneity Structure of complex molecules Assessment of the biological properties constitutes an equally essential step in establishing a complete characterization profile. An important property is the biological activity that describes the specific ability or capacity of a product to achieve a defined biological effect. A valid biological assay to measure the biological activity should be provided by the manufacturer. Examples of procedures used to measure biological activity include: � Animal-based biological assays, which measure an organism's biological response to the product; � Cell culture-based biological assays, which measure biochemical or physiological response at the cellular level; � Biochemical assays, which measure biological activities such as enzymatic reaction rates or biological responses induced by immunological interactions. Other procedures such as ligand and receptor binding assays, may be acceptable. Potency (expressed in units) is the quantitative measure of biological activity based on the attribute of the product which is linked to the relevant biological properties, whereas, quantity (expressed in mass) is a physicochemical measure of protein content. Mimicking the biological activity in the clinical situation is not always necessary. A correlation between the expected clinical response and the activity in the biological assay should be established in pharmacodynamic or clinical studies. The results of biological assays should be expressed in units of activity calibrated against an international or national reference standard, when available and appropriate for the assay utilized. Where no such reference standard exists, a characterized in-house reference material should be established and assay results of production lots reported as in-house units. Often, for complex molecules, the physicochemical information may be extensive but unable to confirm the higher-order structure which, however, can be inferred from the biological activity. In such cases, a biological assay, with wider confidence limits, may be acceptable when combined with a specific quantitative measure. Importantly, a biological assay to measure the biological activity of the product may be replaced by physicochemical tests only in those instances where: � sufficient physicochemical information about the drug, including higher-order structure, can be thoroughly established by such physicochemical methods, and relevant correlation to biologic activity demonstrated; and � there exists a well-established manufacturing history. Where physicochemical tests alone are used to quantitate the biological activity (based on appropriate correlation), results should be expressed in mass. Test Procedures and Acceptance Criteria for Biotechnological/Biological Products For the purpose of lot release (section 4), the choice of relevant quantitative assay (biological and/or physicochemical) should be justified by the manufacturer. 2.1.3 Immunochemical properties When an antibody is the desired product, its immunological properties should be fully characterized. Binding assays of the antibody to purified antigens and defined regions of antigens should be performed, as feasible, to determine affinity, avidity and immunoreactivity (including cross-reactivity). In addition, the target molecule bearing the relevant epitope should be biochemically defined and the epitope itself defined, when feasible. For some drug substances or drug products, the protein molecule may need to be examined using immunochemical procedures (e.g., ELISA, Western-blot) utilizing antibodies which recognize different epitopes of the protein molecule. Immunochemical properties of a protein may serve to establish its identity, homogeneity or purity, or serve to quantify it. If immunochemical properties constitute lot release criteria, all relevant information pertaining to the antibody should be made available. Assessment of the biological properties constitutes an equally essential step in establishing a complete characterization profile. An important property is the biological activity that describes the specific ability or capacity of a product to achieve a defined biological effect. A valid biological assay to measure the biological activity should be provided by the manufacturer. Examples of procedures used to measure biological activity include: � Animal-based biological assays, which measure an organism's biological response to the product; � Cell culture-based biological assays, which measure biochemical or physiological response at the cellular level; � Biochemical assays, which measure biological activities such as enzymatic reaction rates or biological responses induced by immunological interactions. Other procedures such as ligand and receptor binding assays, may be acceptable. Potency (expressed in units) is the quantitative measure of biological activity based on the attribute of the product which is linked to the relevant biological properties, whereas, quantity (expressed in mass) is a physicochemical measure of protein content. Mimicking the biological activity in the clinical situation is not always necessary. A correlation between the expected clinical response and the activity in the biological assay should be established in pharmacodynamic or clinical studies. The results of biological assays should be expressed in units of activity calibrated against an international or national reference standard, when available and appropriate for the assay utilized. Where no such reference standard exists, a characterized in-house reference material should be established and assay results of production lots reported as in-house units. Often, for complex molecules, the physicochemical information may be extensive but unable to confirm the higher-order structure which, however, can be inferred from the biological activity. In such cases, a biological assay, with wider confidence limits, may be acceptable when combined with a specific quantitative measure. Importantly, a biological assay to measure the biological activity of the product may be replaced by physicochemical tests only in those instances where: � sufficient physicochemical information about the drug, including higher-order structure, can be thoroughly established by such physicochemical methods, and relevant correlation to biologic activity demonstrated; and � there exists a well-established manufacturing history. Where physicochemical tests alone are used to quantitate the biological activity (based on appropriate correlation), results should be expressed in mass. Test Procedures and Acceptance Criteria for Biotechnological/Biological Products For the purpose of lot release (section 4), the choice of relevant quantitative assay (biological and/or physicochemical) should be justified by the manufacturer. 2.1.3 Immunochemical properties When an antibody is the desired product, its immunological properties should be fully characterized. Binding assays of the antibody to purified antigens and defined regions of antigens should be performed, as feasible, to determine affinity, avidity and immunoreactivity (including cross-reactivity). In addition, the target molecule bearing the relevant epitope should be biochemically defined and the epitope itself defined, when feasible. For some drug substances or drug products, the protein molecule may need to be examined using immunochemical procedures (e.g., ELISA, Western-blot) utilizing antibodies which recognize different epitopes of the protein molecule. Immunochemical properties of a protein may serve to establish its identity, homogeneity or purity, or serve to quantify it. If immunochemical properties constitute lot release criteria, all relevant information pertaining to the antibody should be made available.

5. Interferon-gamma

6. Attributes & Combinatorics These changes are not necessarily independent The vast majority of these occur at miniscule levels or not at all They are generally not characterized in many products However some of these may matter. There have been difference in products pot change that have not been correlated with a particular structural attribute. Maybe some of these depend on combinations of changes. Characterization of structure and heterogeneity is not enough-?These changes are not necessarily independent The vast majority of these occur at miniscule levels or not at all They are generally not characterized in many products However some of these may matter. There have been difference in products pot change that have not been correlated with a particular structural attribute. Maybe some of these depend on combinations of changes. Characterization of structure and heterogeneity is not enough-?

7. Removal of glycosylation by mutation of Asn297 eliminates binding to Fc-receptors and C1q , and increases the sensitivity to proteases, though the half-life of IgG1 is not effected (Wright and Morrison, TIBTECH review 1997 vol.15).

8. Types of Specifications Identity & purity should include: native and denatured size (sensitive to aggregates) chargeIdentity & purity should include: native and denatured size (sensitive to aggregates) charge

9. Safety Specifications

10. Potency Assay Types-Bioassay In vivo assays Organ or tissue assays Cell culture assays Cell line or population response Late response (proliferation, cytokines) Early response (signaling pathway) Multiple cell types (MLR, cell-cell adhesion) Biochemical assays catalytic Ab enzyme blockade

11. Potency Assay Continuum

12. Presentation Outline IND Content, Characterization & Specifications for Biologics Manufacturing Changes & Comparability Contaminants in Manufacturing Common Pitfalls

13. How Much of the Iceberg (desired product) Can We See?

14. Comparability The extent of the quality information required is dependent on the clinical/non clinical data set transferring to product produced from the new process Less quality data are used to support comparability assessments in early development Thus communication regarding clinical status is critical to assess comparability data needed

15. Differences are Often Observed Impact of observed differences may be known or judged to be of low risk If impact is unknown, additional testing may be necessary pre-clinical and/or clinical studies Lack of observed differences without appropriate characterization may still require additional testing

16. Changes in Manufacturing Switch from vial to syringe for Cytokine Stability decreased, new impurities were detected following approval Metal ions from stopper activated trace proteases in product Large Scale change in Drug Substance manufacture Small shift in post-translational modifications A significant shift in bioavailability

18. Immunogenicity Erythropoetin Changed to a HSA-free formulation in syringes No changes in quality parameters or Pk but antibody associated cases of pure red cell aplasia Interferon beta 1a New MCB and facility Post changed product had no �significant� differences in physicochemical attributes, bioactivity or PK - product was marketed. Post marketing study; incidence of neutralizing Ab changed from 24 to 5%

19. Comparability Additional Studies

20. Presentation Outline IND Content, Characterization & Specifications for Biologics Manufacturing Changes & Comparability Contaminants in Manufacturing Common Pitfalls

21. Source Materials Currently the Division has over 460 Pre-license files for products in clinical development for a wide range of uses. In addition, we regulate 24 products already on the market, including 19 Biologics License Applications (and an additional 2 that are currently under review), 4 New Drug Applications, and 1 antibody for a cell selection device. The next slide <next slide> shows a list Currently the Division has over 460 Pre-license files for products in clinical development for a wide range of uses. In addition, we regulate 24 products already on the market, including 19 Biologics License Applications (and an additional 2 that are currently under review), 4 New Drug Applications, and 1 antibody for a cell selection device. The next slide <next slide> shows a list

22. Infectious Agents Testing Cell Bank & End of Production Sterility Mycoplasma Virus (Adventitious, Species-specific, Retrovirus) Source material screening Human (HIV, HBV, HCV, CJD, etc.) Animal (TSE sources, viruses) Process Filtrations Environmental controls Viral clearance studies

23. Viral Clearance Endogenous for early dev Adventitious for late dev Evaluate unit operations

24. Alternative Clearance studies Clearance can be used for contaminants as well as virus Generic Clearance Study � virus clearance is demonstrated for steps in the purification of a mAb and extrapolated to another mAb purified by the identical process. Modular Clearance Study � virus clearance is demonstrated on individual purification steps (modules) that � may use different model mAb. Identical modules may be extrapolated to other mAb. Abbreviated Testing for certain products/studies Many caveats in applying some of these approaches For Generic and Modular mAb only applicable to biochemically similar mAb purified by identical methods. Acceptable for homologous products, however only mAb have meet the necessary criteria. Basic fundamental and evaluation of the processes being evaluated Approaches may be acceptable - meet basic tenants of process validation, Consistency, robustness How close do processing conditions have to be? Used in combination with small scale studies Assess all sources of variability, worst case scenarioMany caveats in applying some of these approaches For Generic and Modular mAb only applicable to biochemically similar mAb purified by identical methods. Acceptable for homologous products, however only mAb have meet the necessary criteria. Basic fundamental and evaluation of the processes being evaluated Approaches may be acceptable - meet basic tenants of process validation, Consistency, robustness How close do processing conditions have to be? Used in combination with small scale studies Assess all sources of variability, worst case scenario

25. Types of Impurities Product related impurities Process related impurities Media components (insulin, transferrin) Chemical additives (antibiotics, inducing agents) Leachables (Protein A, resins, heavy metals) Cell components (HCP & DNA)

26. Mab Original Submission Holds

27. Credits Patrick Swann Barry Cherney Kurt Brorson Keith Webber Wendy Shores

28. The End No product development was harmed in the making of these slides

29. 21CFR Information for Investigators 312.23(a)(5) Investigator's brochure. If required .. containing the following information: (i) A brief description of the drug substance and the formulation, including the structural formula, if known. (ii-v) Information related to safety & efficacy 312.55 � The sponsor shall, as the overall investigation proceeds, keep each participating investigator informed of new observations� 312.32(c) IND safety reports �The sponsor shall notify FDA and all participating investigators in a written IND safety report�

30. Guidances 1. Changes to an Approved Application for Specified Biotechnology and Specified Synthetic Biological Products - Final 7/1997 2. Changes to an Approved NDA or ANDA - Final 4/2004 3. Changes to an Approved NDA or ANA; Specifications - Use of Enforcement Discretion for Compendial Changes - Final 11/2004 4. Comparability Protocols -- Chemistry, Manufacturing, and Controls Information- Draft 2/2003 5. Container Closure Systems for Packaging Human Drugs and Biologics � Final 5/1999 6. Demonstration of Comparability of Human Biological Products, Including Therapeutic Biotechnology-derived Products - Final 4/1996 7. Drug Master Files Current DMF Information - Final 9/1989 8. Environmental Assessment of Human Drug and Biologics Applications � Final 7/1998

31. Guidances 9. Format and Content for the CMC Section of an Annual Report - Final 9/1994 10. INDs for Phase 2 and Phase 3 Studies Chemistry, Manufacturing, and Controls Information - Final 5/2003 11. IND Meetings for Human Drugs and Biologics Chemistry, Manufacturing, and Controls Information - Final 5/2001 12. Interpreting Sameness of Monoclonal Antibody Products Under the Orphan Drug Regulations - Draft 7/1999 13. Monoclonal Antibodies Used as Reagents in Drug Manufacturing - Final 3/2001 14. NDAs: Impurities in Drug Substances - Final 2/2000 15. Submission Documentation for Sterilization Process Validation in Applications for Human and Veterinary Drug Products - Final 11/1994

32. ICH Documents Q1A(R2) Stability Testing of New Drug Substances and Products - Final 11/2003 Q1B Photostability Testing of New Drug Substances and Products - Final 11/1996 Q1C Stability Testing for New Dosage Forms - Final 5/9/1997 Q1D Bracketing and Matrixing Designs for Stability Testing �fNew Drug Substances and Products - Final 1/2003 Q1E Evaluation of Stability Data - Final 6/2004 Q2A Text on Validation of Analytical Procedures - Final 3/1995 Q2B Validation of Analytical Procedures: Methodology - Final 5/19/1997 Q3A(R) Impurities in New Drug Substances - Final 2/1 0/2003 . Q3B(R) Impurities in New Drug Products - Final 11/2003 Q3C Impurities: Residual Solvents - Final 12/24/1997

33. ICH Documents Q5B Quality of Biotechnological Products: Analysis of the Expression Construct in Cells Used for Production of-DNA Derived Protein Products - Final 2/1996 Q5C Quality of Biotechnological Products: Stability Testing of Biotechnological/Biological Products - Final 7/1996 Q5D Quality of Biotechnological/Biological Products: Derivation and Characterization of Cell Substrates Used for Production of Biotechnological/Biological Products; Availability - Final 9/21/1998 Q5E Comparability of Biotechnological/iological Products Subject to Changes in their Manufacturing Process - Final 6/2005 Q6A International Conference on Harmonisation; Guidance on Q6A Specifications:Test Procedures and Acceptance Criteria for New Drug Substances and New Drug Products: Chemical Substances - Final 12/29/2000

34. Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products - Final 8/1999 Q7 A Good Manufacturing Practice Guidance for Active Pharmaceutical Ingredients - Final 8/2001 Q8(R2) Pharmaceutical Development - Final 8/2009 Q9 Quality Risk Management - Final 96/1/2006 Q10 Quality Systems - 4/8/2009 ICH Documents

35. Relevant Attributes � those molecular and biological characteristics found to be useful in ensuring the safety and efficacy of the product (ICH Q6B) Defining those attributes is relevant to: Risk management CGMPs for the 21st century Can these attribute be defined? Often difficult Default is to look at many attributes Importance of biological activity by appropriate techniques (which includes the determination of physicochemical properties, biological activity, immunochemical properties, purity and impurities)by appropriate techniques (which includes the determination of physicochemical properties, biological activity, immunochemical properties, purity and impurities)