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Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
Usp  biotherapeutics - biological medicines
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Usp biotherapeutics - biological medicines

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12th USP Science & Standards Symposium - New Delhi

12th USP Science & Standards Symposium - New Delhi

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  • 1. Track II, Session I: Biological Medicines–Biotherapeutics Wednesday, April 17, 2013 (9:00 a.m. to 11:00 a.m.) IPC–USP Science & Standards Symposium Partnering Globally for 21st Century Medicines
  • 2. Moderator: Venkata Ramana, Ph.D. Reliance Life Sciences
  • 3. Monograph Tests for Monoclonal Antibody Therapeutics Venkat Mukku, Ph.D. Consultant, USP
  • 4. Recombinant Therapeutic Mabs approved or in review in the EU and US (as of 2-28-2013) International nonproprietary name Muromonab-CD3 Abciximab Rituximab Basiliximab Daclizumab Palivizumab Infliximab Trastuzumab Gemtuzumab ozogamicin Type Anti-CD3; Murine IgG2a Anti-GPIIb/IIIa; Chimeric IgG1 Fab Anti-CD20; Chimeric IgG1 Anti-IL2R; Chimeric IgG1 Anti-IL2R; Humanized IgG1 Anti-RSV; Humanized IgG1 Anti-TNF; Chimeric IgG1 Anti-HER2; Humanized IgG1 Anti-CD33; Humanized IgG4 Indication first approved Reversal of kidney transplant rejection Prevention of blood clots in angioplasty Non-Hodgkin's lymphoma Prevention of kidney transplant rejection Prevention of kidney transplant rejection Prevention of respiratory syncytial virus infection Crohn disease Breast cancer Acute myeloid leukemia First EU (US) approval year 1986* (1986#) 1995* (1994) 1998 (1997) 1998 (1998) 1999 (1997); # 1999 (1998) 1999 (1998) 2000 (1998) NA (2000#) Alemtuzumab Adalimumab Tositumomab-I131 Efalizumab Cetuximab Ibritumomab tiuxetan Omalizumab Bevacizumab Natalizumab Trade name Orthoclone Okt3 Reopro MabThera, Rituxan Simulect Zenapax Synagis Remicade Herceptin Mylotarg MabCampath, Campath1H Humira Bexxar Raptiva Erbitux Zevalin Xolair Avastin Tysabri Anti-CD52; Humanized IgG1 Anti-TNF; Human IgG1 Anti-CD20; Murine IgG2a Anti-CD11a; Humanized IgG1 Anti-EGFR; Chimeric IgG1 Anti-CD20; Murine IgG1 Anti-IgE; Humanized IgG1 Anti-VEGF; Humanized IgG1 Anti-a4 integrin; Humanized IgG4 Chronic myeloid leukemia Rheumatoid arthritis Non-Hodgkin lymphoma Psoriasis Colorectal cancer Non-Hodgkin's lymphoma Asthma Colorectal cancer Multiple sclerosis 2001 (2001) 2003 (2002) NA (2003) 2004 (2003); # 2004 (2004) 2004 (2002) 2005 (2003) 2005 (2004) 2006 (2004) Ranibizumab Panitumumab Eculizumab Lucentis Vectibix Soliris Anti-VEGF; Humanized IgG1 Fab Anti-EGFR; Human IgG2 Anti-C5; Humanized IgG2/4 Macular degeneration Colorectal cancer Paroxysmal nocturnal hemoglobinuria 2007 (2006) 2007 (2006) 2007 (2007) Certolizumab pegol Golimumab Canakinumab Cimzia Simponi Ilaris Crohn disease 2009 (2008) Rheumatoid and psoriatic arthritis, ankylosing spondylitis 2009 (2009) Muckle-Wells syndrome 2009 (2009) Catumaxomab Ustekinumab Tocilizumab Ofatumumab Denosumab Belimumab Ipilimumab Removab Stelara RoActemra, Actemra Arzerra Prolia Benlysta Yervoy Brentuximab vedotin Pertuzumab Adcetris Perjeta Anti-TNF; Humanized Fab, pegylated Anti-TNF; Human IgG1 Anti-IL1b; Human IgG1 Anti-EPCAM/CD3;Rat/mouse bispecific Mab Anti-IL12/23; Human IgG1 Anti-IL6R; Humanized IgG1 Anti-CD20; Human IgG1 Anti-RANK-L; Human IgG2 Anti-BLyS; Human IgG1 Anti-CTLA-4; Human IgG1 Anti-CD30; Chimeric IgG1; immunoconjugate Anti-HER2; humanized IgG1 Raxibacumab (Pending) Trastuzumab emtansine Kadcyla Vedolizumab (Pending) Anti-B. anthrasis PA; Human IgG1 Anti-HER2; humanized IgG1; immunoconjugate Anti-alpha4beta7 integrin; humanized IgG1 Source: Janice M. Reichert, Editor-in-Chief, Mabs Malignant ascites Psoriasis Rheumatoid arthritis Chronic lymphocytic leukemia Bone Loss Systemic lupus erythematosus Metastatic melanoma 2009 (NA) 2009 (2009) 2009 (2010) 2010 (2009) 2010 (2010) 2011 (2011) 2011 (2011) Hodgkin lymphoma Breast Cancer 2012 (2011) 2013 (2012) Anthrax infection NA (2012) Breast cancer 2013 Ulcerative colitis, Crohn disease In review (NA)
  • 5. Mabs ranked in the Top Ten Drugs in 2012 • • • • • • • • • • Humira (AbbVie) - Mab Advair (Glaxo Smith Kline) Rituxan (Roche) - Mab Lantus (Sanofi) Herceptin (Roche) - Mab Crestor (AstraZeneca) Remicade (Johnson & Johnson) - Mab Avastin (Roche) - Mab Cymbalta (Eli Lilly and Co) Plavix (Sanofi & Bristol-Myers Squibb)
  • 6. Developing Standards for Mabs Challenges in developing Mab standards • Structure is complex, often heterogeneous, with a mix of isoforms, glycoforms etc. • Need to understand the importance of each form • Many types of impurities, requiring different types of analytical methods. Impurities often are manufacturing process-dependent • Potency assay (Bioassay) is a key quality attribute. Multiple potency assays are needed to address different functional domains
  • 7. IgG Structure and Functional Domains Variable Region CH1 VL CL Constant Region CH2 Fc • Effector Functions ADCC (FcgRI, II, III) CDC (C1q) • FcRn binding CH3 Multiple domains lead to multiple variants requiring a battery of physicochemical and biological tests
  • 8. Mab Variants / Isoforms • • • • • • • • • N-terminal Pyro-Glu Iso-Asp formation Deamidation Methionine oxidation Heavy Chain Cterminal Lysine Heavy chain C-terminal Prolinamide variants Disulfide Variants Free Cys adducts Glycosylation variants – High mannose, G0, G1, G2 – Fucosylation – Sialic acid capping CH1 VL CL CH2 CH3 Not all variants impact safety and/or efficacy
  • 9. Characterization Tests for Mabs Most of these tests are not used in routine testing, but are important for detailed understanding of the product and Comparability / Biosimilarity • Primary Structure – LC/MS Peptide Maps, 2nd enzyme – N-terminal Sequence – C-terminal Prolinamide formation – Disulfide Bond Determination – Intact Mass Determination – Amino Acid Analysis (AAA)  Others – DSC – AUC – SEC-MALS – Extinction Coefficient determination – Excipients methods • Secondary and Tertiary Structure – FTIR – Far UV CD – Near UV CD – Intrinsic fluorescence  Functional characterization tests – Additional cell-based assays – Antigen binding – Effector functions – Fcg RI, RIII binding – FcRn binding
  • 10. Quality Tests for Mabs Quality Attribute Identity Test Bioassay, Peptide map, cIEF Function mediated by antigen binding site Potency Fc-mediated effector functions Antigen binding Purity: Glycosylation variants Glycan profiling: Oligosaccharide mapping Product -Related Impurities High molecular weight. impurities Size Exclusion Chromatography , CE-SDS (Reduced) Low molecular weight impurities CE-SDS under non-reducing conditions (CE-SDS-NR) Charge heterogeneity Cation exchange chromatography (CEX) Methionine oxidation Protein-A HPLC or Peptide map LC-MS Strength (protein concentration) A280 nm Process-Related Impurities Host Cell Proteins, DNA, Protein-A Compendial tests pH, endotoxin, osmolality, microbial tests, excipients, particulates, visual tests
  • 11. Potency Assays for Mabs Depend upon the mechanism of action (MoA) • Anti-Proliferation, cytostatic effects • Antibody-toxin Mediated Cytotoxicity • Complement dependent cytotoxicity (CDC) • Antibody Dependent Cell-mediated Cytotoxicity assay (ADCC) • Apoptosis • Inhibit inflammatory cytokine activity or secretion • Etc.
  • 12. Complement Dependent Cytotoxicity (CDC) Target B-Cell Activation of Complement Cascade Rituximab C1q Dead cell subsequent to CDC
  • 13. ADCC Activity Antibody Dependent Cell-Mediated Cytotoxicity Target B-Cell Cytotoxic mediators Dead B-Cell upon ADCC Rituximab Effector Cell Fc Receptor RI, RIII
  • 14. Potency Assays for Rituximab CDC assay Cell line: WIL-2S Cell viability measured by AlamarBlue Readout: fluorescence ADCC assay Target cells: WIL2-S Effector cells: Jurkat cell engineered to express FcgRIII and NFAT-luciferase reporter gene; Readout: luminescence
  • 15. CDC-Based Potency Assay for Rituximab Acceptable Procedure USP Reference Procedure Cell line and bioassay: CDC assay using CD20+ cells CDC assay using WIL-2S cells, human complement and Resazurin-based detection reagent Data Analysis: Relative potency (RP) calculated according to appropriate data analysis guidelines Relative potency calculated per USP <1034> and validated per USP <1033> Specificity: Unrelated Mab and Formulation buffer should not show activity. Unrelated Mab and Formulation buffer showed no activity. Precision (Repeatability): RSD ≤ 10% RSD 5% Precision (Intermediate precision): RSD ≤ 15% RSD 9% Linearity: r2 > 0.9 measured and expected potency r2 1.0 Accuracy: Spike recovery 85%–115% Range: Minimally cover 80%-125%; wider is better 95%–105% 64% to 156%
  • 16. Peptide Maps for Identity of Mabs
  • 17. Peptide Maps for Identity of Mabs Rituximab Bevacizumab Trastuzumab Tryptic peptide maps showing differences between 3 Mabs
  • 18. cIEF as an Identity Test for Mab
  • 19. Role of Fc Glycans in Mab Functions Terminal galactose is important for CDC N-Acetyl glucosamine essential for ADCC Bisecting N-Acetyl glucosamine enhances ADCC Core fucose inhibitory to ADCC; afucosylated variants show higher ADCC High-mannose glycans reduce half life of IgG 120 Degalactosylation abolishes 40% of CDC activity. % Potency by CDC assay • • • • • 100 80 60 40 20 0 Treatment with Beta-Galactosidase
  • 20. Glycan Profiling of Mabs Deglycosylation with PNGase followed by 2-Aminobenzamide labeling and Normal Phase UPLC. Peaks are characterized using 2AB-Labeled glycan standards 2.50 G1F’ G1F G0F 3.00 c c 2.00 G0 N-acetyl glucosamine G2F Core Fucose Galactose Sialic Acid G2FS1 G2 G1’ G1 Man 5 0.50 G0F-GN 1.00 G2FS2 1.50 G0-GN EU Mannose 0.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 Minutes 20.00 22.00 24.00 26.00 28.00 30.0
  • 21. N- & C-terminal Charge Variants of Mabs E E E pE pE pE pE pE E pE VH CH1 VL CL CH2 S L S L S P G K S L S L S P G K Un-Processed SS L L SS L L SS PP GG K Partially Processed S S L L S S L L S S PP GG K CH3 Partially Processed E = N-terminal glutamate; pE = N-terminal pyroglutamate; K= C-terminal lysine Other charge variants: Deamidation, glycation, terminal sialic acid S L S L S P G S L S L S P G Fully Processed
  • 22. Degradation-Related Charge Variants of Mabs
  • 23. CEX Chromatography of Rituximab Before Carboxy Peptidase treatment After Carboxy Peptidase treatment
  • 24. Size Exclusion Chromatography Retuximab standard Retuximab heat treated to generate HMW impurities
  • 25. Capillary Electrophoresis-SDS (CE-EDS) Non-reducing conditions (CE-SDS-NR) to quantitate low molecular weight impurities Reducing conditions (CESD-R) to quantitate NonGlycosylated Heavy Chain and
  • 26. Biosimilar Guideline in Korea and ASEAN Countries Mr. Bonjoong Kim, Celltrion, Inc.
  • 27. CONTENTS SECTION I. SECTION II. Overview On Global Biosimilar Guidelines KOREA & ASEAN Regulatory Environments SECTION III. Biosimilar Guidelines in KOREA SECTION IV. Biosimilar Guidelines in ASEAN SECTION V. Executive Summary
  • 28. SECTION I Overview On Global Biosimilar Guidelines
  • 29. Global Regulation or Guideline on Biosimilar [SUMMARY]  EU: In fact, world first well-organized biosimilar legislation and guideline were implemented in EU.  KOREA: Biosimilar guidelines have been issued since first biosimilar guideline was implemented in 2009.  ASEAN: Malaysia and Singapore are leading implementation of biosimilar guidelines in ASEAN. 2011 SNG 2012 2013 USA EGY India
  • 30. Global Regulation or Guideline on Biosimilar [SUMMARY]  Biosimilar Pathway: EU, Brazil, Argentina,Turkey, Malaysia, Canada, Mexico, Korea, Japan, India, Singapore, Egypt, Saudi Arabia, Australia, US, Venezuela  Biosimilar Pathway under Development: Peru, Chile, Ecuador, Colombia, Bolivia, Paraguay, Uruguay, Pakistan, South Africa, New Zealand, Philippines, Hong Kong, Vietnam, Indonesia, Thailand Biosimilar Pathways Biosimilar Pathways under Development No Biosimilar Pathway
  • 31. SECTION II KOREA & ASEAN Regulatory Environments
  • 32. Guideline on Biosimilar in KOREA & ASEAN [SUMMARY]  Korea, Singapore and Malaysia has their own biosimilar pathway.  Other ASEAN countries does not have official biosimilar pathway yet. CLASSIFICATION COUNTRY Biosimilar Pathways (1+2) Biosimilar Pathways under Development (4) Biosimilar Pathways under Development No Biosimilar Pathway Philippines, Vietnam, Indonesia, Thailand No Biosimilar Pathway (4) Biosimilar Pathways Korea, Malaysia, Singapore Laos, Myanmar, Cambodia, Brunei Darussalam
  • 33. Regulatory Environment in Korea & ASEAN [SUMMARY]  Regulatory Agency in Indonesia and Thailand has tendency to treat biosimilar filing as new biologics with less requirements  Regulatory Agency in Indonesia and Thailand has tendency to treat biosimilar filing as generic pathway with more requirements Regulatory Environment COUNTRY Biosimilar Pathway Korea, Malaysia, Singapore New biologics (with less requirement) Indonesia, Thailand Generic Pathway Philippines, Vietnam Unknown Laos, Myanmar, Cambodia, Brunei Darussalam
  • 34. Regulatory Environment in Korea & ASEAN [SUMMARY]  KOREA: Regulatory Agency is being changed with its name and function.  Philippines: Regulatory Agency is being changed with new PFDA commissioner.  Thailand and Malaysia: GMP Inspection or Accreditation is time-consuming. COUNTRY Regulatory Environment Korea Name and organization of Korea FDA was changed in March 2013. Name: KFDA  MDFS (Ministry of Drug and Food Safety) Organization and Function: Being changed to extend its function. Singapore EU or US CPP is required. Malaysia Long waiting list for GMP Inspection. Thailand TFDA is now preparing biosimilar guideline GMP Accreditation Process was implemented. Philippines Organization of PFDA is being changed with new commissioner. Indonesia Local Manufacturing is required. Vietnam Special Quota VISA & Valid VISA are available. Laos, Myanmar, Cambodia, Brunei Darussalam Not known.
  • 35. SECTION III Biosimilar Guidelines in KOREA
  • 36. Guideline on Biosimilar in KOREA <Reference: EMA Guidelines> mAb Nonclinical Clinical
  • 37. Guideline on Biosimilar in KOREA [SUMMARY]  KOREA has similar structure with EU biosimilar guideline  Legislation: Regulations on Review & Authorization of Biological Products (KFDA Notification No. 2009-59)  Guideline: 8 relevant guidelines and 1 Q&A are now available. Definition/Principle General Guidelines Quality, Safety, Efficacy Product Specific Requirement
  • 38. Guideline on Biosimilar in KOREA Overarching Guideline Guidelines on the Evaluation of Biosimilar Products (2009) SCOPE In principle: Biosimilar concept applicable to any biological medicinal product. In practice: Only for products that can be thoroughly characterised. [Choice of Reference Product] Korea The reference product should be authorized in Korea. However, if the registered reference product is not available in Korea commercially, the same biological product as the one authorized in Korea (including the manufacturing site and the manufacturing process) may be purchased from overseas markets and used as the reference product. EU (Reference Information) Must be authorised in the EU based on a complete dossier.
  • 39. Guideline on Biosimilar in KOREA Overarching Guideline Guidelines on the Evaluation of Biosimilar Products (2009) [Quality Requirement] Korea A complete description of the manufacturing process for the drug substance and drug product should be provided in detail including information on quality control/quality assurance, inprocess controls, and process validation. Comparability study along with specifications, analytical procedures and stability study should be conducted. EU (Reference Information) For recombinant proteins, an extensive comparability exercise is required. Quality aspects of comparability must be considered in relation to implications for safety and efficacy. Purity and impurity profiles of the active substance and medicinal product must be assessed qualitatively and quantitatively for the biosimilar and the reference product.
  • 40. Guideline on Biosimilar in KOREA Overarching Guideline Guidelines on the Evaluation of Biosimilar Products (2009) [Nonclinical Data] Korea EU (Reference Information) ▶ „ In vitro studies: Assays, such as receptor-binding studies or cellbased assays should normally be undertaken in order to establish the comparability of the biological/ pharmacodynamic activity of the biosimilar product and the reference product, should be conducted Non-clinical studies for biosimilar versions of recombinant proteins should be comparative in nature, designed to detect differences in response between the biosimilar and the reference product. In vitro studies, in vivo studies in relevant animal species, and at least one repeat-dose toxicity study in relevant animal species normally should be conducted. ▶ „ In vivo studies: - Animal studies should be performed in species know to be relevant - At least one repeat-dose toxicity study - Other toxicological studies, including safety pharmacology, reproductive toxicology, mutagenicity and carcinogenicity studies are not required for biosimilar unless warranted by the results from repeated toxicological studies.
  • 41. Guideline on Biosimilar in KOREA Overarching Guideline Guidelines on the Evaluation of Biosimilar Products (2009) [Clinical Data] Korea EU (Reference Information) The clinical comparability studies include pharmacokinetic, pharmacodynamic, and efficacy studies. If the comparability can be demonstrated by confirmatory pharmacokinetic/pharmacodyna mic data, an efficacy study may be omitted. For recombinant proteins, comparative efficacy clinical trials usually are necessary to demonstrate clinical comparability. The clinical requirements depend on the existing knowledge about the reference product and the claimed therapeutic indication(s). For recombinant proteins, a biosimilar’s immunogenicity must always be investigated. Immunogenicity risks in different indications should be considered separately.
  • 42. Guideline on Biosimilar in KOREA Overarching Guideline Guidelines on the Evaluation of Biosimilar Products (2009) [Extrapolation of Indication] Korea EU (Reference Information) Extrapolation of these data to other indications of the reference product for which post-marketing survey was completed may be possible if all of the following conditions are fulfilled: For recombinant proteins, in certain cases, it may be possible to extrapolate therapeutic similarity to other indications. Justification of extrapolation depends on clinical experience, available literature, whether the same mechanism of action or receptor is involved in both indications, and possible safety issues in different subpopulations.  A sensitive clinical test model has been used that is able to detect potential differences between the biosimilar product and the reference product;  The clinically relevant mechanism of action and/or involved receptor(s) are the same;  Safety and immunogenicity have been sufficiently characterized.
  • 43. Guideline on Biosimilar in KOREA Overarching Guideline Guidelines on the Evaluation of Biosimilar Products (2009) [PV and RMP] Korea Safety data obtained from clinical trials are usually sufficient for product authorization, but further close monitoring of clinical safety of the biosimilar product is usually necessary in the post-marketing phase. EU (Reference Information) By law, a risk management plan or pharmacovigilance plan must be submitted for biosimilars as for other medicines. The plan should take into account risks identified during product development and potential risks and how those risks will be addressed after authorisation For recombinant proteins, clinical safety must be monitored closely after authorisation. The application should include a risk specification and pharmacovigilance plan.
  • 44. SECTION IV Biosimilar Guidelines in ASEAN
  • 45. Guideline on Biosimilar in ASEAN – SINGAPORE & MALAYSIA SINGAPORE MALAYSIA GUIDANCE ON MEDICINAL PRODUCT REGISTRATION IN SINGAPORE GUIDANCE DOCUMENT FOR APPLICANTS : (APPENDIX 17 GUIDANCE ON REGISTRATION OF SIMILAR BIOLOGICAL PRODUCTS IN SINGAPORE) INFORMATION AND SUBMISSION REQUIREMENTS FOR REGISTRATION OF BIOSIMILARS [Choice of Reference Product] SINGAPORE The reference product must be registered in Singapore. MALAYSIA EU (Reference Information) The reference product must Must be authorised in the be registered in Malaysia. EU based on a complete dossier.
  • 46. Guideline on Biosimilar in ASEAN – SINGAPORE & MALAYSIA SINGAPORE MALAYSIA GUIDANCE ON MEDICINAL PRODUCT REGISTRATION IN SINGAPORE GUIDANCE DOCUMENT FOR APPLICANTS : (APPENDIX 17 GUIDANCE ON REGISTRATION OF SIMILAR BIOLOGICAL PRODUCTS IN SINGAPORE) INFORMATION AND SUBMISSION REQUIREMENTS FOR REGISTRATION OF BIOSIMILARS [Quality Requirement] SINGAPORE MALAYSIA EU (Reference Information) Comparability data between the biosimilar product and the reference product (in terms of quality) must be submitted in the quality dossier. The extent of the comparability studies and the assessment criteria depends on the complexity of the product and the capability of the methods used to demonstrate comparability. The comparability exercise should entail evaluation of both drug substance and drug product. A full quality dossier is always required. In addition, extensive data focused on the similarity, including comprehensive side-byside physicochemical and biological characterisation of the biosimilar and the reference product should be submitted For recombinant proteins, an extensive comparability exercise is required. Quality aspects of comparability must be considered in relation to implications for safety and efficacy. Purity and impurity profiles of the active substance and medicinal product must be assessed qualitatively and quantitatively for the biosimilar and the reference product.
  • 47. Guideline on Biosimilar in ASEAN – SINGAPORE & MALAYSIA SINGAPORE MALAYSIA GUIDANCE ON MEDICINAL PRODUCT REGISTRATION IN SINGAPORE GUIDANCE DOCUMENT FOR APPLICANTS : (APPENDIX 17 GUIDANCE ON REGISTRATION OF SIMILAR BIOLOGICAL PRODUCTS IN SINGAPORE) INFORMATION AND SUBMISSION REQUIREMENTS FOR REGISTRATION OF BIOSIMILARS [Nonclinical Data] SINGAPORE MALAYSIA EU (Reference Information) In vitro studies: Assays like receptor-binding studies or cell-based assays should normally be undertaken in order to establish comparability in reactivity and the likely causative factor(s) if comparability cannot be established. Animal studies should be performed to investigate pharmacodynamic effect/ activity relevant to the clinical application, non-clinical toxicity as determined in at least one repeat dose toxicity study, including toxicokinetic measurements, and specific safety concerns. Safety pharmacology, reproduction toxicology, mutagenicity and carcinogenicity studies are not required for biosimilar products, unless indicated by the results of repeat dose studies. ▶ „ In vitro studies: Receptor-binding studies or cellbased assay should be conducted ▶ „ In vivo studies: - Animal pharmacodynamic study where approriate, relevant to clinical use - At least one repeat-dose toxicity study Other toxicological studies, including safety pharmacology, reproductive toxicology, mutagenicity and carcinogenicity studies are not required for biosimilar unless warranted by the results from repeated toxicological studies. Non-clinical studies for biosimilar versions of recombinant proteins should be comparative in nature, designed to detect differences in response between the biosimilar and the reference product. In vitro studies, in vivo studies in relevant animal species, and at least one repeat-dose toxicity study in relevant animal species normally should be conducted.
  • 48. Guideline on Biosimilar in ASEAN – SINGAPORE & MALAYSIA SINGAPORE MALAYSIA GUIDANCE ON MEDICINAL PRODUCT REGISTRATION IN SINGAPORE GUIDANCE DOCUMENT FOR APPLICANTS : (APPENDIX 17 GUIDANCE ON REGISTRATION OF SIMILAR BIOLOGICAL PRODUCTS IN SINGAPORE) INFORMATION AND SUBMISSION REQUIREMENTS FOR REGISTRATION OF BIOSIMILARS [Clinical Data] SINGAPORE The clinical comparability exercise should begin with pharmacokinetic (PK) and pharmacodynamic (PD) studies followed by clinical efficacy and safety studies. Comparative PK studies designed to demonstrate clinical comparability between the biosimilar product and the reference product with regard to key PK parameters are required. Pharmacodynamic studies to demonstrate therapeutic efficacy of the product is required. The immunogenicity of a biosimilar product must always be investigated. The assessment of immunogenicity requires an optimal antibody testing strategy, characterisation of the observed immune response, as well as evaluation of the correlation between antibodies and pharmacokinetics or pharmacodynamics, relevant for clinical safety and efficacy in all aspects. MALAYSIA EU (Reference Information) Comparative pharmacokinetic studies should be conducted to demonstrate the similarities in pharmacokinetic (PK) characteristics between biosimilar and the reference product. Clinical efficacy trials should also be coundcuted. Other PK studies such as interaction studies or other special populations (e.g children, elderly, patients with renal or hepatic insufficiency) are usually not required. Studies should demonstrate clinical sfaety and immunogenicity of the drug product. For recombinant proteins, comparative efficacy clinical trials usually are necessary to demonstrate clinical comparability. The clinical requirements depend on the existing knowledge about the reference product and the claimed therapeutic indication(s). For recombinant proteins, a biosimilar’s immunogenicity must always be investigated. Immunogenicity risks in different indications should be considered separately.
  • 49. Guideline on Biosimilar in ASEAN – SINGAPORE & MALAYSIA SINGAPORE MALAYSIA GUIDANCE ON MEDICINAL PRODUCT REGISTRATION IN SINGAPORE GUIDANCE DOCUMENT FOR APPLICANTS : (APPENDIX 17 GUIDANCE ON REGISTRATION OF SIMILAR BIOLOGICAL PRODUCTS IN SINGAPORE) INFORMATION AND SUBMISSION REQUIREMENTS FOR REGISTRATION OF BIOSIMILARS [Extrapolation of Indication] SINGAPORE MALAYSIA EU (Reference Information) In case the reference medicinal product has more than one indication, the efficacy and safety of the biosimilar product has to be justified or, if necessary, demonstrated separately for each of the claimed indications. In certain cases it may be possible to extrapolate therapeutic similarity shown in one indication to other indications of the reference medicinal product. Justification will depend on e.g., clinical experience, available literature data, whether or not the same mechanisms of action or the same receptor(s) are involved in all indications. Possible safety issues in different subpopulations should also be addressed. Demonstration of similarity may also allow extrapolation of efficacy and safety data to other indications of the reference product. For recombinant proteins, in certain cases, it may be possible to extrapolate therapeutic similarity to other indications. Justification of extrapolation depends on clinical experience, available literature, whether the same mechanism of action or receptor is involved in both indications, and possible safety issues in different subpopulations.
  • 50. Guideline on Biosimilar in ASEAN – SINGAPORE & MALAYSIA SINGAPORE MALAYSIA GUIDANCE ON MEDICINAL PRODUCT REGISTRATION IN SINGAPORE GUIDANCE DOCUMENT FOR APPLICANTS : (APPENDIX 17 GUIDANCE ON REGISTRATION OF SIMILAR BIOLOGICAL PRODUCTS IN SINGAPORE) INFORMATION AND SUBMISSION REQUIREMENTS FOR REGISTRATION OF BIOSIMILARS [PV&PMP] SINGAPORE MALAYSIA EU (Reference Information) ADR reporting, PSURs, Risk management plan, Educational Materials and Product Sales Data for the biosimilar product is required The pharmacovigilance, as part of a comprehensive RMP, should include regular testing for consistent manufacturing of the biosimilar. The pharmacovigilance plan must be approved prior to approval of product and the system must be in place to conduct monitoring. By law, a risk management plan or pharmacovigilance plan must be submitted for biosimilars as for other medicines. The plan should take into account risks identified during product development and potential risks and how those risks will be addressed after authorisation. For recombinant proteins, clinical safety must be monitored closely after authorisation. The application should include a risk specification and pharmacovigilance plan.
  • 51. Guideline on Biosimilar in ASEAN – SINGAPORE & MALAYSIA SINGAPORE MALAYSIA GUIDANCE ON MEDICINAL PRODUCT REGISTRATION IN SINGAPORE GUIDANCE DOCUMENT FOR APPLICANTS : (APPENDIX 17 GUIDANCE ON REGISTRATION OF SIMILAR BIOLOGICAL PRODUCTS IN SINGAPORE) INFORMATION AND SUBMISSION REQUIREMENTS FOR REGISTRATION OF BIOSIMILARS [PV&PMP] SINGAPORE MALAYSIA EU (Reference Information) A product is interchangeable with another if both products are approved for the same indication, and can be used for the said indication. Two products are substitutable with each other if they can both be used in lieu of the other during the same treatment period. For interchangeable products, one or the other can be used (prescribed) but these products cannot be substituted with one another during a treatment period. Interchangeability does not imply substitutability. Unlike generic chemical drugs, whereby the chemical structure is identical to that of the reference chemical product, a biosimilar product does not usually have an identical structure to the reference biological product. Therefore, even though a biosimilar product may be approved to be similar in terms of quality, safety and efficacy to the reference product, immunogenicity may preclude switching between products. Given current science, biosimilars cannot be considered interchangeable with the reference product or products of the same class. Substitution is determined at the member state level, and therefore this topic is not directly addressed in EMA guidance. EMA guidance states that biosimilars are not generic medicinal products and that the decision to treat a patient with a reference product or a biosimilar should be made following the opinion of a qualified healthcare professional.
  • 52. SECTION IV Executive Summary
  • 53. Executive Summary – Regulation & Guideline <Regulation & Guideline> Topic Reference EU US JAPAN Directive 2001/83/EC The Biologics Price Competition and Innovation Act of 2009 (sections 7001 to 7003 of the Patient Protection and Affordable Care Act, Pub. L. 111-148 (2010)). Ministry of Health Labor and Welfare, Pharmaceutical and Food Safety Bureau, Guideline for Ensuring Quality, Safety, and Efficacy for Biosimilar Products, PFSB/ELD Notification No. 0304007. ▶ „ Guideline on Similar Biological Medicinal Products EMEA/CHMP/437/04. ▶ „ Guideline on Similar Biological Medicinal Products Containing BiotechnologyDerived Proteins as Active Substance: Nonclinical and Clinical Issues EMEA/CHMP/BMWP/42832/20 05. ▶ „ Guideline on Similar Biological Medicinal Products Containing BiotechnologyDerived Proteins as Active Substance: Quality Issues EMEA/CHMP/BWP/49348/2005 . ▶ „ Guideline on Immunogenicity Assessment of Biotechnology-Derived Therapeutic Proteins EMEA/CHMP/BMWP/14327/06. ▶ Formal Meetings Between the FDA and Biosimilar Biological Product Sponsors or Applicants ▶ Quality Considerations in demonstrating biosimilarity to a reference protein product ▶ Questions and Answers ▶ Scientific Considerations in demonstrating biosimilarity to a reference product Law (Regulation) Guideline WHO KOREA ASEAN MALAYSIA Regulations on Review & Authorization of Biological Products (KFDA Notification No. 2009-59) WHO, Guidelines on Evaluation of Similar Biotherapeutic Products (SBPs). SINGAPORE ▶ Medicines Act (Chapter 176) ▶ Poisons Act (Chapter 234) ▶ Misuse of Drugs Regulations – subsidiary legislation under the Misuse of Drugs Act (Chapter 185) ▶ Sale of Drugs Act (Chapter 282) ▶ Medicines (Advertisement and Sale) Act (Chapter 177) Control of Drugs and Cosmetic Regulations 1984 (CDCR 1984) Guidelines on the Evaluation of Biosimilar Products GUIDANCE ON MEDICINAL PRODUCT REGISTRATION IN SINGAPORE - APPENDIX 17 GUIDANCE DOCUMENT FOR APPLICANTS : INFORMATION AND SUBMISSION REQUIREMENTS FOR REGISTRATION OF BIOSIMILARS
  • 54. Executive Summary – Reference Product <Reference Product> Topic Reference EU Must be authorised in the EU based on a complete dossier. Reference Product US Must be licensed under a full biologics licence application (under section 351(a) of the Public Health Service Act). JAPAN Must be authorized in Japan. WHO Should be authorised in the country or region in question (or, where the licensing country lacks an authorised reference product, should be authorised and widely marketed in another jurisdiction with a wellestablished regulatory framework for and experience in evaluation and post-market surveillance of biotherapeutics). KOREA The reference product should be authorized in Korea. However, if the registered reference product is not available in Korea commercially, the same biological product as the one authorized in Korea (including the manufacturing site and the manufacturing process) may be purchased from overseas markets and used as the reference product. ASEAN SINGAPORE The reference product must be registered in Singapore MALAYSIA The reference product must be registered in Malaysia
  • 55. Executive Summary – Quality <Quality> Topic Quality Requirement Reference EU US For recombinant proteins, an extensive comparability exercise is required. Quality aspects of comparability must be considered in relation to implications for safety and efficacy. Purity and impurity profiles of the active substance and medicinal product must be assessed qualitatively and quantitatively for the biosimilar and the reference product. Unless the Secretary makes a determination that it is unnecessary, an application must contain data from analytical studies demonstrating that the biosimilar is highly similar to the reference product (notwithstanding minor differences in clinically inactive components). JAPAN A biosimilar must be fully characterised, including by conducting studies comparing the structure and composition, physicochemical properties, bioactivity, and immunologic properties of the biosimilar against its reference product. WHO Comprehensive physicochemical and biological characterisation of the biosimilar in head-to-head comparisons with the reference product is required, and all aspects of quality and heterogeneity should be assessed. KOREA A complete description of the manufacturing process for the drug substance and drug product should be provided in detail including information on quality control/quality assurance, in-process controls, and process validation. Comparability study along with specifications, analytical procedures and stability study should be conducted. ASEAN SINGAPORE MALAYSIA Comparability data between the biosimilar product and the reference product (in terms of quality) must be submitted in the quality dossier. The extent of the comparability studies and the assessment criteria depends on the complexity of the product and the capability of the methods used to demonstrate comparability. The comparability exercise should entail evaluation of both drug substance and drug product. A full quality dossier is always required. In adiition, extensive data focused on the similarity, including comprehensive side-by-side physicochemical and biological characterisation of the biosimilar and the reference product should be submitted.
  • 56. Executive Summary – Nonclinical Data <Nonclinical Data> Topic Nonclinical Data Reference EU US JAPAN WHO Non-clinical studies for biosimilar versions of recombinant proteins should be comparative in nature, designed to detect differences in response between the biosimilar and the reference product. In vitro studies, in vivo studies in relevant animal species, and at least one repeat-dose toxicity study in relevant animal species normally should be conducted. Unless the Secretary makes a determination that it is unnecessary, an application must include data derived from animal studies (including the assessment of toxicity) to help demonstrate that the product is biosimilar to a reference product. It is important to understand the limitations of such animal studies (e.g., small sample size, intraspecies variations) when interpreting results comparing the proposed product and the reference product. A sponsor may be able to provide a scientific justification for a stand-alone toxicology study using only the proposed product instead of a comparative toxicology study. For a more detailed discussion on the design of animal toxicology studies, see ICH S6/S6(R1). Before performing clinical studies, the biosimilar applicant must conduct nonclinical studies to verify that the product can be safely administered to humans. Before conducting nonclinical studies, a biosimilar must be subjected to a full quality characterisation. The pharmacological action of the biosimilar and its reference product should be compared through nonclinical pharmacological studies, and repeat-dose toxicity and toxicokinetic studies may be useful. The non-clinical evaluation should include pharmacodynamic, pharmacokinetic, and comparative repeat-dose toxicity studies in a relevant species. The amount of additional non-clinical data required is dependent on product-specific factors. In vitro assays like receptor binding studies or cell-based assays should normally be conducted to establish comparability of pharmacodynamic activity. Animal studies in a relevant species should generally be conducted. KOREA ▶ „ In vitro studies: Assays, such as receptorbinding studies or cellbased assays should normally be undertaken in order to establish the comparability of the biological/ pharmacodynamic activity of the biosimilar product and the reference product. should be conducted ▶ „ In vivo studies: - Animal studies should be performed in species know to be relevant - At least one repeatdose toxicity study - Other toxicological studies, including safety pharmacology, reproductive toxicology, mutagenicity and carcinogenicity studies are not required for biosimilar unless warranted by the results from repeated toxicological studies. ASEAN SINGAPORE MALAYSIA In vitro studies: Assays like receptor-binding studies or cell-based assays should normally be undertaken in order to establish comparability in reactivity and the likely causative factor(s) if comparability cannot be established. Animal studies should be performed to investigate pharmacodynamic effect/ activity relevant to the clinical application, nonclinical toxicity as determined in at least one repeat dose toxicity study, including toxicokinetic measurements, and specific safety concerns. Safety pharmacology, reproduction toxicology, mutagenicity and carcinogenicity studies are not required for biosimilar products, unless indicated by the results of repeat dose studies. ▶ „ In vitro studies: Receptor-binding studies or cell-based assay should be conducted ▶ „ In vivo studies: - Animal pharmacodynamic study where approriate, relevant to clinical use - At least one repeat-dose toxicity study Other toxicological studies, including safety pharmacology, reproductive toxicology, mutagenicity and carcinogenicity studies are not required for biosimilar unless warranted by the results from repeated toxicological studies.
  • 57. Executive Summary – Clinical Data <Clinical Data> Topic Clinical Data Reference EU US JAPAN WHO For recombinant proteins, comparative efficacy clinical trials usually are necessary to demonstrate clinical comparability. The clinical requirements depend on the existing knowledge about the reference product and the claimed therapeutic indication(s). For recombinant proteins, a biosimilar’s immunogenicity must always be investigated. Immunogenicity risks in different indications should be considered separately. Unless the Secretary makes a determination that it is unnecessary, an application must include data derived from a clinical study or studies (including the assessment of immunogenicity and pharmacokinetics or pharmacodynamics) that are sufficient to demonstrate safety, purity, and potency in one or more appropriate conditions of use for which the reference product is licensed and intended to be used. The purpose of these data is to help demonstrate that the product is biosimilar to a reference product. Clinical studies should generally be required, but they may not be required where nonclinical data are sufficient to assure bioequivalence and quality equivalence (if bioequivalence and quality equivalence have been demonstrated). Pharmacokinetic or pharmacodynamic study results are, however, inconclusive concerning clinical efficacy. Clinical studies should be conducted to verify efficacy for the specific indications for which the biosimilar applicant seeks authorisation. Clinical studies should be designed to demonstrate comparable safety and efficacy of the biosimilar and reference product. Clinical trials are required to demonstrate similar efficacy. Immunogenicity should always be investigated in humans before authorisation. KOREA The clinical comparability studies include pharmacokinetic, pharmacodynamic, and efficacy studies. If the comparability can be demonstrated by confirmatory pharmacokinetic/phar macodynamic data, an efficacy study may be omitted. ASEAN SINGAPORE MALAYSIA The clinical comparability exercise should begin with pharmacokinetic (PK) and pharmacodynamic (PD) studies followed by clinical efficacy and safety studies. Comparative PK studies designed to demonstrate clinical comparability between the biosimilar product and the reference product with regard to key PK parameters are required. Pharmacodynamic studies to demonstrate therapeutic efficacy of the product is required. The immunogenicity of a biosimilar product must always be investigated. The assessment of immunogenicity requires an optimal antibody testing strategy, characterisation of the observed immune response, as well as evaluation of the correlation between antibodies and pharmacokinetics or pharmacodynamics, relevant for clinical safety and efficacy in all aspects. Comparative pharmacokinetic studies should be conducted to demonstrate the similarities in pharmacokinetic (PK) characteristics between biosimilar and the reference product. Clinical efficacy trials should also be coundcuted. Other PK studies such as interaction studies or other special populations (e.g children, elderly, patients with renal or hepatic insufficiency) are usually not required. Studies should demonstrate clinical sfaety and immunogenicity of the drug product.
  • 58. Executive Summary – Extrapolation of Indication <Extrapolation of Indication> Topic Extrapolation of Indication Reference EU US JAPAN WHO For recombinant proteins, in certain cases, it may be possible to extrapolate therapeutic similarity to other indications. Justification of extrapolation depends on clinical experience, available literature, whether the same mechanism of action or receptor is involved in both indications, and possible safety issues in different subpopulations. The sponsor will need to provide sufficient scientific justification for extrapolating clinical data to support a determination of biosimilarity for each condition of use for which licensure is sought. Such scientific justification should address, for example, the following issues for the tested and extrapolated conditions of use. The MOA(s) in each condition of use for which licensure is sought. The PK and biodistribution of the product in different patient populations; PD measures may provide important information on the MOA. Extrapolation may be permitted if the mechanism of action is not unclear, the applicant can show that a similar pharmacological result can be expected for the relevant indications, and the mechanism of action does not differ among indications. Extrapolation may be possible if a sensitive clinical test model has been used that is able to detect potential differences between the products, that the mechanism of action and/or receptors are the same, and that the safety and immunogenicity of the biosimilar have been characterised and there are no special safety issues expected with the extrapolated indication. KOREA Extrapolation of these data to other indications of the reference product for which postmarketing survey was completed may be possible if all of the following conditions are fulfilled: - A sensitive clinical test model has been used that is able to detect potential differences between the biosimilar product and the reference product; - The clinically relevant mechanism of action and/or involved receptor(s) are the same; - Safety and immunogenicity have been sufficiently characterized. ASEAN SINGAPORE MALAYSIA In case the reference medicinal product has more than one indication, the efficacy and safety of the biosimilar product has to be justified or, if necessary, demonstrated separately for each of the claimed indications. In certain cases it may be possible to extrapolate therapeutic similarity shown in one indication to other indications of the reference medicinal product. Justification will depend on e.g., clinical experience, available literature data, whether or not the same mechanisms of action or the same receptor(s) are involved in all indications. Possible safety issues in different subpopulations should also be addressed. Demonstration of similarity may also allow extrapolation of efficacy and safety data to other indications of the reference product.
  • 59. Executive Summary – Naming <Naming> Topic Naming Reference EU To support pharmacovigilance monitoring, the specific medicinal product given to the patient should be clearly identified. The name, appearance, and packaging of a biosimilar medicine should differ from those of the reference product. US Not addressed. JAPAN WHO Notification PFSB/EKD No. 0304011 regulates the naming of biosimilars and states that the nonproprietary names and proprietary names of biosimilars should be readily distinguishable from the names of reference products and other biosimilars. For nonproprietary names, the following should be added to the end of the non-proprietary name: Follow-on 1 [2, 3, and so on]. For proprietary names, the letters BS should be added to the end of the name, along with the dosage form, dosage, and name of the manufacturer. Biosimilars should be clearly identifiable by a unique proprietary name. Where an international nonproprietary name (INN) is defined, that name should be stated and the WHO’s policy on INNs should be followed. KOREA Not Adressed ASEAN SINGAPORE MALAYSIA Not Adressed All biosimilars should be distinguishable by name i.e assign a brand name explicitly, using names that are not suggestive towards the originator nor towards other biosimilars.
  • 60. Executive Summary – Labeling <Labeling> Topic Reference EU Not addressed. Labeling US Not addressed. JAPAN Not addressed. WHO The prescribing information for the biosimilar should be as similar as possible to that of the reference product, except for product-specific information, such as different excipient(s). If the biosimilar has fewer indications than the reference product, the information related to those indications may be omitted unless it is considered important to inform doctors and patients about certain risks. In these cases, the prescribing information should clearly state that the biosimilar is not indicated for use in the specific indication(s) and the reasons for it. A national regulatory authority may choose to require prescribing information to mention the product is a biosimilar, discuss the studies performed with the biosimilar, and/or include instructions to the physician on how to use the biosimilar. KOREA Not Adressed ASEAN SINGAPORE MALAYSIA Not Adressed It is expected that the labeling of biosimilar meet the following criteria: - A clear indication that the medicine is a biosimilar of a specific reference product - The invented name, common or scientific name and the manufacturer’ name s - Clinical data for the biosimilar describing the clinical similarity (i.e safety and efficacy) to the reference product and in which indication(s) - Interchangeability and substitution advice should clearly and prominently state that the biosimilar is not interchangeable or substitutable with the reference product.
  • 61. Executive Summary – Pharmacovigilance and risk <Pharmacovigilance and risk> Topic Pharmaco -vigilance and risk Reference EU US JAPAN WHO By law, a risk management plan or pharmacovigilance plan must be submitted for biosimilars as for other medicines. The plan should take into account risks identified during product development and potential risks and how those risks will be addressed after authorisation For recombinant proteins, clinical safety must be monitored closely after authorisation. The application should include a risk specification and pharmacovigilance plan. The Secretary’s existing authority to require a risk evaluation and mitigation strategy (REMS) for drugs applies to biosimilars (a REMS documents requirements designed to minimise risk associated with a drug). In addition, the Secretary’s existing authority to mandate post-market studies and clinical trials as well as post-market labelling changes applies to biosimilars. A post-marketing safety surveillance plan and a risk management plan for biosimilars is required and should be submitted with the application. A pharmacovigilance plan is required when an application is submitted and a risk management plan may be necessary in some cases. KOREA Safety data obtained from clinical trials are usually sufficient for product authorization, but further close monitoring of clinical safety of the biosimilar product is usually necessary in the post-marketing phase. ASEAN SINGAPORE MALAYSIA ADR reporting, PSURs, Risk management plan, Educational Materials and Product Sales Data for the biosimilar product is required. The pharmacovigilance, as part of a comprehensive RMP, should include regular testing for consistent manufacturing of the biosimilar. The pharmacovigilance plan must be approved prior to approval of product and the system must be in place to conduct monitoring.
  • 62. Executive Summary – Interchangeability and substitution <Interchangeability and substitution> Topic Interchangeability and substitution Reference EU US Substitution is determined at the member state level, and therefore this topic is not directly addressed in EMA guidance. EMA guidance states that biosimilars are not generic medicinal products and that the decision to treat a patient with a reference product or a biosimilar should be made following the opinion of a qualified healthcare professional. Substitution is determined at state level in accordance with state pharmacy laws. FDA must find a biosimilar to be interchangeable with its reference product if the information submitted by the biosimilar applicant demonstrates that: ▶ „ The applicant’s product is biosimilar to the reference product (under the law’s standard for biosimilarity). ▶ „ The applicant’s product can be expected to produce the same clinical result as the reference product in any given patient. For products administered more than once to an individual, the applicant must also demonstrate that the risk in terms of safety or diminished efficacy of alternating or switching between use of the biosimilar and use of the reference product is not greater than the risk of using the reference product without such alternation or switch. JAPAN WHO Substitution of a Not biosimilar for, or addressed. the combined use of a biosimilar with, a reference product should be avoided during the postmarketing surveillance period. KOREA ASEAN SINGAPORE Not A product is interchangeable Addressed with another if both products are approved for the same indication, and can be used for the said indication. Two products are substitutable with each other if they can both be used in lieu of the other during the same treatment period. For interchangeable products, one or the other can be used (prescribed) but these products cannot be substituted with one another during a treatment period. Interchangeability does not imply substitutability. Unlike generic chemical drugs, whereby the chemical structure is identical to that of the reference chemical product, a biosimilar product does not usually have an identical structure to the reference biological product. Therefore, even though a biosimilar product may be approved to be similar in terms of quality, safety and efficacy to the reference product, immunogenicity may preclude switching between products. A warning statement on the risks associated with switching of products during treatment, and against product substitution, is to be included in the package insert of the biosimilar product. MALAYSIA Given current science, biosimilars cannot be considered interchangeable with the reference product or products of the same class.
  • 63. Executive Summary – Data protection for originators <Data protection for originators> Topic Data protection for originators Reference EU US JAPAN WHO For a reference product for which the application for approval was submitted in October or November 2005 or later (different rules apply to a reference product for which the application for approval was submitted earlier), a biosimilar may not be marketed until ten years after authorisation of its reference product that is a new active substance. This period may be extended for an additional year, if within eight years after reference product authorisation, the reference product is authorised for a new therapeutic indication that constitutes a significant clinical benefit in comparison with existing therapies. A biosimilar may not be approved until 12 years after the date on which the reference product was first licensed under section 351(a) of the Public Health Service Act. The date of first licensure does not include the date of approval of a supplement to the reference product application or the date of approval of a subsequent application for either: „A change to the reference product, other than a structural change, that results in a new indication, route of administration, dosing schedule, dosage form, delivery system, delivery device, or strength. „A structural modification to the reference product that does not result in a change in safety, purity, or potency. A biosimilar application cannot be approved until the innovative product on which the application relies has completed an eight-year reexamination or post-marketing surveillance period. Not Addressed KOREA Not Addressed ASEAN SINGAPORE MALAYSIA Not Addressed Not Addressed
  • 64. Biosimilar Guideline in Latin American Countries Mr. Daniel Guedes Bio-Manguinhos / FIOCRUZ / Brazil
  • 65. Oswaldo Cruz Foundation Fiocruz / Brazil Bio-Manguinhos Institute of Technology in Immunobiologicals
  • 66. Biosimilar Guideline in Latin American Countries FIOCRUZ / Bio-Manguinhos Institute of Technology in Immunobiologicals Vaccines Reactives Biopharmaceuticals
  • 67. Biosimilar Guideline in Latin American Countries FIOCRUZ / Bio-Manguinhos Vaccines  Diphtheria, Tetanus, Pertussis and Hib;  Yelow Fever;  Haemophilus influenzae b;  Meningitis A e C;  Poliomyelitis;  Measles, Mumps and Rubella.  Tetravalent viral (SmallPox).
  • 68. Biosimilar Guideline in Latin American Countries FIOCRUZ / Bio-Manguinhos Reactives Molecular Tests NAT HIV/HCV; Rapid Tests DPP HIV, Syphilis, Canine Leishmaniasis, Leptospirosis;
  • 69. Biosimilar Guideline in Latin American Countries Reactives Parasitological Tests Sorological Tests Kato-Katz Imunoenzimatic C. Leishmaniasis Ind. Immunofluorescence C. Leishmaniasis H. Leishmaniasis Chagas Disease
  • 70. Biosimilar Guideline in Latin American Countries Biopharmaceuticals Alphapoetin Alphainterferon
  • 71. Biosimilar Guideline in Latin American Countries Daniel Guedes, Ph.D. Quality Control Department FIOCRUZ / Bio-Manguinhos - Brazil
  • 72. Latin American Countries Pharmacopoeia Argentina Farmacopoea Argentina Brazil Farmacopéia Brasileira Chile Chilean Pharmacopoeia Foundation México Farmacopea de los Estados Unidos Mexicanos
  • 73. NATIONAL HEALTH SURVEILLANCE AGENCY COLLEGIATE BOARD – ANVISA (BRAZIL)  Integration of Latin American countries Pharmacopeias;  Optimize the use of resources and knowledge; Mercosur`s Pharmacopoeia
  • 74. Biosimilar Guideline in Latin American Countries Paraguai Argentina Uruguai Brazil
  • 75. Biosimilar Guideline in Latin American Countries Venezuela Suriname Peru Equador
  • 76. Biosimilar Guideline in Latin American Countries Guidelines from Argentina Law 16.643 ANMAT 7075/2011 Biologicals 3397/2012 Biotechnologicals 7729/2011 Biosimilars 1490/1992
  • 77. Biosimilar Guideline in Latin American Countries Guidelines from Argentina Law 16.643 “Are subject to this law and regulations, import, export, production, preparation, processing, commercialization or deposit in national jurisdiction or interprovincial trade of drugs, chemicals, reagents, pharmaceutical forms, diagnostic elements, and all other product use and application in human medicine and people whether natural or involving in such activities. All this products shall meet the conditions set in Argentina`s Pharmacopoeia.”
  • 78. Biosimilar Guideline in Latin American Countries Guidelines from Argentina Rule 7075/2011 Establish the conditions and requirements for registration of medical specialties with biological origin.
  • 79. Biosimilar Guideline in Latin American Countries Guidelines from Argentina Rule 3397/2012 Authorization Biotechnology products  Monoclonal antibodies;  Recombinant products;
  • 80. Biosimilar Guideline in Latin American Countries Guidelines from Argentina Rule 7729/2011 – Authorization Biosimilars  Medical specialty of biological origin with previous marketing.  Comparability referential product;  Characterization with modern methods;  Similar behavior to the referential product;
  • 81. Biosimilar Guideline in Latin American Countries NATIONAL HEALTH SURVEILLANCE AGENCY COLLEGIATE BOARD – ANVISA (BRAZIL) RDC 46 / 2000 RDC 55 / 2010 RDC 49 / 2011 RDC 61 / 2008 RDC 50 / 2011 RDC 17 / 2010 RDC 233 / 2011
  • 82. Biosimilar Guideline in Latin American Countries RDC 46/ 2000 Regulates production processes and quality control, acquisition and distribution of medicines for human use.
  • 83. Biosimilar Guideline in Latin American Countries RDC 61/ 2008 Provides on Criteria for Harmonization of Nomenclature of Vaccines and Serums.
  • 84. Biosimilar Guideline in Latin American Countries RDC 17/ 2010 Requirements for the Good Manufacturing Practice in Medicinal Products for human
  • 85. Biosimilar Guideline in Latin American Countries RDC 55 / 2010 Provides on the registration of new biological products (Biosimilar) and biological products.
  • 86. Biosimilar Guideline in Latin American Countries RDC 55, Scope I. Vaccines; II. Hyper-immune serums; III. Hemoderivatives; IV. Biodrugs (Biologicals or by Biotechnology); V. Monoclonal antibodies; VI. Drugs (live, attenuated or dead microorganisms).
  • 87. Biosimilar Guideline in Latin American Countries RDC 49 / 2011 Provides on post-registration changes and inclusions, suspension and reactivation of manufacturing and cancellations of biological product registration.
  • 88. Biosimilar Guideline in Latin American Countries RDC 50 / 2011 Stipulates the procedures and conditions for conducting stability studies for registration or changes post-registration of biological products.
  • 89. Biosimilar Guideline in Latin American Countries RDC 233 / 2011 Technical Regulation of Production and Quality Control for Registration, Changing PostRegistration and Revalidation of Allergen Extracts and Products.
  • 90. Biosimilar Guideline in Latin American Countries Conclusions  The WHO Biosimilar Guidelines influenced Latin America;  The structure is similar to those of USP;  There are new but consistent Guidelines for Biosimilars;  More countries can join the group.
  • 91. Biosimilar Guideline in Latin American Countries
  • 92. Impact of Product Variants on Product Functional Activity and Stability in the Content of Biosimilars Anita Krishnan, Ph.D., Lupin Limited
  • 93. Overview of Product and Process inter-relationship Acceptable ranges for a subset of these QAs are normally established based on a combination of clinical experience, pre-clinical studies, lab studies and prior knowledge. The acceptable ranges are used to establish the boundaries for the design spaces in the Upstream, Downstream and Drug Products sections of the case study.
  • 94. Critical Quality Attributes “A CQA is a physical, chemical, biological, or microbiological property or characteristic that should be within an appropriate limit, range, or distribution to ensure the desired product quality. Desired product quality includes clinically safe and efficacious product.” ICH Q8 (R2) Product-Related Substances • Glycosylation variants (Galactose; Man5; Sialic Acid; Afucose) • Acidic Variants (Deamidated/Sialylated Species) • Oxidation • Unpaired cysteine residues • C-terminal Lysine Variants 10 Aggregation & Clips Glycosylation Product-Related Impurities • Clips / Truncations • Aggregates 5 Oxidation Process-Related Impurities • Residual Protein A (Affinity Resin) • Host Cell Proteins • DNA • Endotoxin/Bioburden/Particulates/Viruses 1 Deamidation Q cyclization Lys variants In the case of Biosimilars most of the CQAs are available as prior art.
  • 95. CQAs related to Fab variations Deamidation Glycation CQAs related to Fc effector functions Gal content and CDC Afucose and ADCC FcRn and PK
  • 96. IgG Sequence analysis for Fab CQAs Asn 30 to Asp CDR 1 CDR 2 Fab glycosylation : N-X-S/T CDR 3 K glycation Fab related variations Asn 55 to Asp CDR 1 CDR 2 CDR 3 Asp 102 to isoAsp
  • 97. CQAs Related to Fab Variations Asp30 isoAsp102 R. J. Harris et al. / J. Chromatogr. B 752 (2001) 233 –245 Reaction of primary amines of lysine residues with reducing sugars such as glucose/lactose to form aldimine (Schiff’s base ) and later a ketoamine adduct
  • 98. Molecular Signatures of Fc effector Functions Fcɣ R I, II, III FcRn - Met256 Fcɣ R I, II, III HINGE C1q C1q FcRn – His protonation DP Deamidation
  • 99. Mechanistic pathways of IgG Rituximab Trastuzumab Cetuximab Infliximab Golimumab Adalimumab Omalizumab Bevacizumab
  • 100. Role of neonatal Fc receptor (FcRn) in PK Regulates systemic half-life by salvaging FcRn bound IgG and recycling IgG:FcRn interaction depends on conserved His in CH2-CH3 interface Salt bridges between positive residues of IgG and negative amino acids on FcRn surface
  • 101. Case Study of In-house Batch Glycan profile : NP - HPLC 0.275 0.275 0.250 0.250 G0F 0.225 0.225 G1F/G1F’ 0.200 0.200 RMP 0.175 In-house batch 0.150 G2F G1 +Man5 G0 0.125 0.150 300 G2 LU LU 0.175 0.125 0.100 0.100 250 0.075 0.075 Charge variants : IE - HPLC Potency : CDC activity 0.050 0.050 200 0.025 0.025 7000 0.000 Ref std B6197 B06 In house Sample 0.000 38 40 42 44 46 48 50 52 54 56 58 60 62 6000 64 66 68 70 72 74 Minutes 76 78 80 150 5000 In-house batch mAU 36 4000 100 3000 RMP 2000 50 1000 0 0 0.01 Potency: 67.5% CI: 91.3% - 109.0% 0.1 1 10
  • 102. Impact of Fc glycosylation heterogeneity on effector functions
  • 103. Does ADCC and CDC co-relate with the glycan profile?
  • 104. Structure function relationship
  • 105. Optimized Process: 10L Bioreactor Mannosylation Galactosylation CDC: RMP INIT OPT RMP Afucosylation ADCC: 131% RMP OPT 96 % 92 % INIT OPT Fucosylation 270% INIT G0F G1F G1’F G2F R O I R O I R O I R O I
  • 106. Oxidation of rHu mAb – reduced binding to Protein A 30000 uV 25000 20000 15000 Oxidized impurities 10000 mAb-RMP In-house 1 In-house 2 5000 0 5.0 10.0 15.0 20.0 25.0 30.0 5% TBHP 1% TBHP 3% TBHP 0.5% TBHP Oxidized Native 35.0 min
  • 107. Site Specific Oxidation of rHu mAbs on chemical stress MWD: Signal A, 214 nm/Bw:4 nm Rituximab RMP Ctrl Rituximab RMP Ctrl.dat 1200 MWD: Signal A, 214 nm/Bw:4 nm MWD: Signal A, 214 nm/Bw:4 nm MWD: Signal A, 214 nm/Bw:4 nm MWD: Signal A, 214 nm/Bw:4 nm rituximab rmp 0.5 a rituximab rmp 0.25_a rituximab rmp 0.125_a rituximab rmp 1_a 1200 Resolution (DAB) 1000 1000 37°C CTRL 1.0% t-BHP 0.5% t-BHP 0.25% t-BHP 0.125% t-BHP LC 800 Fc/2 Fd 600 600 400 400 Ox Fc/2 200 200 0 0 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 mAU 800
  • 108. Site Specific Oxidation of in-house samples MWD: Signal A, 214 nm/Bw:4 nm dp5 500 mgper50ml_3m at_a 1200 MWD: Signal A, 214 nm/Bw:4 nm dp6 100 mgper10ml_3m rt_a RMP Inhouse 1 Inhouse 2 1000 LC 800 Fc/2 Fd mAU 600 400 Ox Fc/2 200 Ox Fd 0 Oxidation of methionines in Fc and Fd -200 27 28 29 30 31 32 33 Minutes 34 35 36 37 38 39 40 41 42 43
  • 109. Correlation of Oxidation levels and CDC activity of RMP 120.0 % Potency 100.0 y = -0.7676x + 102.53 R² = 0.9291 80.0 60.0 40.0 20.0 0.0 0.0 20.0 40.0 60.0 80.0 100.0 120.0 % Oxidation Sample RMP 5.0% t-BHP RMP 1.0% t-BHP RMP 0.5% t-BHP RMP 0.25% t-BHP RMP 0.125% t-BHP 10% spike of 100% oxidised RMP RMP 37°C CTRL Inhouse 1 Inhouse 2 % Oxidation % Potency 100.0 60.1 44.2 30.2 17.3 10.0 1.6 18.2 59.9 82.3 81.5 83.7 92.8 97.1 3.7 6.1 69.1 62.3
  • 110. Risk based approach for prevention of Oxidation • Enhanced oxidation due to presence of dissolved oxygen • Metal-catalyzed oxidation is site-specific - Copper induced catalysis observed (Baker et al 1978) • Photo-oxidation of Met can form persulfoxide/ Met sulfoxide • Oxidation due to compromised thermal stability
  • 111. Dissolved Oxygen (%) Dissolved oxygen induces oxidative damage Nitrogen purging reduces dissolved oxygen Before After Storage for 12h DO after 12h 96% 0% Open to air 25% 97% 0% Headspace with Nitrogen 2%
  • 112. Tertiary structure – A role in potency (CDC and ADCC)
  • 113. Structure based sequence alignment of the C1q complex components D270 C1q binding epicentre Stretch of charged residues P331 K322 P329 Proline sandwich W87 W110 Van der Waals contacts Hydrogen bonds NATURE |VOL 406 | 20 JULY 2000
  • 114. Site directed mutagenesis for enhanced activity Enhanced ADCC (Increased FcɣRIII a binding) IgG1 Fc CH2 C1q binding Site Enhanced CDC (Increased C1q binding) Sugar Chain CH3 FcɣRIII binding Site FcRn binding Site Improved PK (Increased FcRn binding)
  • 115. Critical Attributes to be assessed for Biosimilars Post-translational Modifications POTENCY EFFICACY IMMUNOGENICITY SAFETY 3 D structure Protein aggregates Three properties of therapeutic proteins – in the opinion of FDA cannot be sufficiently measured but are deemed to be important for understanding the behaviour of protein drugs: post-translational modifications, three-dimensional structures and protein aggregation.
  • 116. Conclusions • Glycosylation has a direct overbearing effect on potency (ADCC and CDC) • Oxidation of Met 256 and 432 have been proved to have a role in FcRn binding and therefore PK • Oxidation of Met has a definite role in CDC activity too • Reasons for reduced potency could be due to multiple factors namely Oxidation, Tertiary structure differences and Reduced CD20 binding • Risk based approach to control oxidation levels and introduction of stability and release methods for oxidation in mAbs
  • 117. New USP Bioassay Chapters Tina S. Morris, Ph.D., Vice President, Biologics & Biotechnology, USP
  • 118. USP Bioassay Chapters Official since 1950:  <111>: Design and Analysis of Biological Assays – A major revision will be proposed in Pharmacopeial Forum PF39(4) The new suite:  <1030>: Biological Assay Chapters – Overview and Glossary – Will be official with the 2nd Supplement of USP36  <1032>: Design and Development of Biological Assays  <1033>: Validation of Biological Assays  <1034>: Analysis of Biological Assays Official All of these chapters are focused on relative potency bioassays. Chapter numbering is important…
  • 119. From General to Specific – Biological Potency <1030> Overview of Bioassay <1032> Development of Biological Assays <1034> Analysis of Biological Assays <111> Design and Analysis of Biological Assays <121> Insulin Assays Insulin Monograph <1033> Validation of Biological Assays Guidance & Information General Requirement Product-Specific Requirement Product Quality Attributes
  • 120. <1032> Design and Development of Biological Assays  Focuses on design and development, rather than analysis  Covers animal assays, cellular assays, enzyme assays and immunoassays  Focuses on design elements, theory and practice – e.g., describes good and bad plate designs  Addresses activities throughout the product life cycle  Some key ideas – – – – fitness for use, different applications of assay biological and statistical assay fundamentals assay from design and development through validation and performance monitoring system and sample suitability
  • 121. <1032> Design and Development of Biological Assays Specific points:  Identify and minimize sources of variability in a method  Record cell line information, before banking, including... – – – – – – – –  identity morphology purity cryopreservation thaw and culture conditions thaw viability growth characteristics functional stability Prepare two-tiered bank: Master and Working – the size of the banks depends on the growth characteristics of the cells, the number of cells required for each assay, and how often the assay will be performed
  • 122. Chapter <1032>’s Recommendations for Standards  Where possible, a Standard should be prepared using the same manufacturing process as the drug substance  Storage conditions may vary from drug substance or product: – Temperature (e.g., –70 °C or –20 °C instead of 2–8 °C) – Container (e.g., plastic vials instead of syringes) – Formulation (e.g., lyophilized formulation or addition of carrier proteins)  Test Standard for stability at appropriate intervals  An initial Standard = Primary Standard Subsequent Standards = Working Standards – Separate SOPs usually required – Trend charts may be useful in identifying the cause of assay drift
  • 123. <1033> Validation of Biological Assays  Describes approaches for validation design (sample selection and replication strategy), validation acceptance criteria, data analysis and interpretation, and bioassay performance monitoring through quality control  Validation parameters discussed include relative accuracy, specificity, intermediate precision, and range – Laboratories may use dilutional linearity to verify the relative accuracy and range of the method – Although robustness is not a requirement for validation, <1033> recommends that a bioassay’s robustness be assessed either prevalidation or with additional experiments soon after validation  Some differences from common practice: – Equivalence testing – Confidence intervals to demonstrate conformance to acceptance criteria
  • 124. <1034> Analysis of Biological Assays  Intended to be useful for biologists and statisticians alike but heavier on the statistics side  Covers models for different types of bioassays – linear and nonlinear – parallel line and slope ratio – Quantal (e.g., yes/no assays)  Develop and elaborate on appropriate uses of equivalence assessment; e.g. for parallelism/similarity  Have also developed a 4PL Bioassay Simulator that lives on the www.usp.org/usp-nf/compendial-tools website and is freely available to the public. Other simulators are in development – Please provide feedback on your needs!
  • 125. Plan for Revisions to <111>  Include only components that USP intends to be required or that are commonly needed in monographs: – Outliers – Combination of independent assays – Confidence Intervals  Revision is on track for publication in PF39(4) issue, appearing in July-August 2013, with public comment period closing September 30, 2013
  • 126. Product-Specific Potency Assays  Can be called out in a Monograph or General Chapter  Monograph requirements supersede Chapter Requirements  A potency test or at least bioidentity test based on a functional assay is required for most biologics and biotechnology-derived medicines licensed for the US market. A few smaller proteins/peptides have only HPLC-based Assays to determine potency (e.g., vasopressin, oxytocin, leuprolide, etc.)
  • 127. Europe and the US Differ in Bioassay Requirements  In Europe, for peptides and small proteins that are considered “well-characterized” neither a unit-based quantitative bioassay nor a bioidentity test are required. This is not the case in the US.  Some examples: – Insulin(s) - US requires bioidentity based on USP <121> Insulin Assay: rabbit blood glucose test – Somatropin - US requires rat weight gain bioidentity test – Glucagon - US requires primary rat liver cell bioidentity assay, see USP <123>Glucagon Bioidentity Tests
  • 128. Compendial Implications Many bioassays and bioidentity tests for licensed biologics are still animal-based Associated concerns:  Assays with generally high variability and poor precision  Challenges in unit assignments and maintenance, especially in relationship to the International Unit where it exists  Ethical issues regarding animal use
  • 129. USP Initiative: Replacement of Animal Assays  Current: <126> Somatropin Bioidentity Tests – USP is evaluating a cell-based assay using a nonproprietary cell line available from commercial sources. The chapter will contain the animal test and the cell assay.  Future: – <123> Glucagon Bioidentity Tests – USP is working with a sponsor to add a cell-based assay to this chapter and distribute the necessary cell line as a Reference Standard – <121> Insulin Assays – USP has reached out to sponsors for suitable cell-based assay for insulins and hopes to revise the chapter with one or more in the future
  • 130. Case Study: Somatropin Bioidentity Test  Official in the Somatropin monographs since 2005 for bioidentity (to assure potency of the material now assigned by an HPLC Assay method; previously the rat method was used to assign potency)  Currently official method uses hypophysectomized rats injected with somatropin for 10 days. Body weight change is measured and potency calculated relative to the USP Somatropin RS  Two sponsors came forward with new cell-based methods. – Both validated and accepted by FDA for use with US-approved products – Bridging to the animal methods was submitted by the sponsors – Verification of method performance and suitability performed in USP laboratory – Most robust method was chosen for development of a new chapter <126>Somatropin Bioidentity Tests, using commercially available, nonproprietary cell line
  • 131. Cell-based Bioidentity Test Format  Uses the cell line Nb2-11 derived from a rat lymphoma  Proliferation of the cells is dependent on factors such as prolactin, IL-2, or hGH  The method is based on measurement of ATP production in response to Somatropin (rhGH) treatment relative to the USP Somatropin RS. The amount of intracellular ATP is directly proportional to the proliferation of the cells due to the hGH treatment.
  • 132. Cell-based Bioidentity Test Format  On the day of the assay, cells are harvested, washed, and seeded in 96-well assay plates in a minimal medium containing 1% horse serum for one hour.  A dilution series of either Somatropin test sample or USP Somatropin RS (or positive and negative controls) is delivered and the cells are incubated for ~30 hours  A luminescent cell substrate solution is added for ~30 min then the luminescence in the wells is measured  Relative potency is calculated using parallel line analysis relative to the USP Somatropin RS  To pass bioidentity, a somatropin sample must possess not less than 2 USP Somatropin Units/mg
  • 133. Representative Data from Cell-based Method* USP Somatropin Dose Response Triplicate Plates Passage #10 Concentration (ng/mL) Col 2 Col 3 Col 4 Col 5 Col 6 Col 7 Col 8 Col 9 Col 10 MEAN STD RSD 10 4185 4513 4577 4731 4564 4834 4667 4619 4586.042 191.4806 4.18% 1 4074 4127 4064 4279 4061 4132 4266 4180 4176 4151.037 81.8658 1.97% 0.5 4506 4494 4566 4536 4434 4492 4450 4541 4551 4507.593 45.3449 1.01% 0.25 3934 3994 4299 4180 4126 4113 4109 4143 3979 4097.593 113.0542 2.76% 0.125 3238 3284 3398 3258 3301 3343 3346 3441 3558 3352.111 101.0542 3.01% 0.0625 2892 2625 2709 2622 2671 2710 2666 2585 2711 2688 88.4498 3.29% 0.03125 2088 2135 2169 2145 2057 2151 2119 2243 2277 2153.815 69.5885 3.23% 0.015625 1540 1586 1611 1595 1505 1586 1523 1555 1627 1569.852 41.1275 2.62% 0.0078125 1162 1230 1283 1222 1210 1284 1249 1296 1257 1243.741 42.5375 3.42% 0 804 861 884 886 918 968 906 897 890.375 47.0213 5.28% S-N Ratio 5.15 3.08% Luminescence Values Somatropin Dose-Response Curve y = 826.73ln(x) + 5094 R² = 0.9919 0.001 0.01 0.1 Somatropin Concentration in ng/mL 5000 4500 4000 3500 3000 2500 2000 1500 1000 500 0 1 *Data courtesy of Michele Virjee, USP Laboratories.
  • 134. Data Summary and Next Steps  The cell-based method has good signal-to-noise ratios, averaging between 5 and 6  Percent RSD’s were generally below 5%.  Little plate-to-plate variability  Plate uniformity analyses performed in the USP laboratory suggested a slight edge effect in the left-most column and upper-most rows of the microtiter plates, but based on the analyses there was no significant impact on the results. The sponsor did not see any uniformity issues.  The sponsor has successfully used the Nb2-11 cells for the procedure through 50 passages. The USP confirmed the method consistency through passage 16 (which was 11 passages post-thaw).
  • 135. Data Summary and Next Steps, cont’d.  Overall, the cell-based method provides a more precise, consistent, and reliable method when compared to the in vivo method  The new chapter <126>Somatropin Bioidentity Tests will be proposed in the PF39(5) issue, appearing September 2013. Public comments are open through November 30, 2013  Collaborative laboratory testing of the USP Somatropin RS for this new purpose will occur in parallel and be complete prior to the chapter moving to official
  • 136. Epoetin – The Commutability Challenge  For epoetin special challenges exist in the transition from in vivo to in vitro assays  Different types of in vitro assays have varying degrees of sensitivity to and correlation with glycosylation (esp. sialylation) differences  In vivo to in vitro transition, equivalence determination and unit transition has to occur on an assay-by-assay basis
  • 137. <124> Epoetin Bioassays – What are the Options?  Option 1: include both in vivo and in vitro assays – In vivo assay: include only normocythemic assay – Potential misuse of in vitro assay to inappropriately assign units of activity to EPO preparations. – Which in vitro assay to include? The proposed cell assay is not the actual assay used for product release  Option 2: include only the in vivo assay with guidance on – How the standard can be used in an in vitro assay (acceptance criteria for in vitro to in vivo activity ratios) – How to assign an in vitro unitage to the USP standard, so that the in vitro assay can be used to assign units to EPO preparations  Both options assume that the monograph refers to chapter <124> for assay requirements
  • 138. Path Forward and Future Opportunities  Recommendation for a Path Forward – Move forward with option 2, and publish <124> in PF, with in the in vivo assay only. This will provide strict guidance on the use of both in vivo and in vitro bioassays to measure the biological activity of erythropoietin (EPO) and will preserve the traceability to the WHO International Standard for EPO – In a second phase, develop a cell-based assay to be included in the chapter. Other cell assays can be used provided that these assays are validated against compendial assays – Physico-chemical assays that can predict outcome of bioassays can also be added in the future.
  • 139. Expert Panel Discussions – Chapter <124>  USP EPO for bioassay RS calibrated against 3rd WHO International Standard for EPO by in vivo mouse bioassay – assigned unitage in International Units (IU) of EPO  Calibration strategy means that USP EPO for bioassay RS can be used directly in in vivo mouse bioassay for calibration of any process-specific EPO preparation.  An in vitro bioassay cannot be used to transfer the unitage from the USP EPO for bioassay RS to a process-specific EPO preparation – Unless you demonstrate that the USP EPO for bioassay RS and the process-specific EPO preparation exhibit an equivalent ratio of in vitro to in vivo potency.
  • 140. Expert Panel Recommendations – Chapter <124>  The ratio of in vitro to in vivo activity is generally inversely correlated with the degree of terminal sialylation, thus highly sialyated products have a higher level of in vivo activity and as such, have a relatively lower ratio of in vitro to in vivo activity.  Different manufacturing processes mean that the degree of terminal sialylation can be highly variable from one EPO preparation to another. – To use an in vitro assay to measure the biological activity of a preparation of EPO, you must have a full understanding of the relationship between its in vivo and in vitro activity. • Include with the USP EPO for bioassay RS, in both the in vivo assay and in a specific in vitro assay.
  • 141. Expert Panel Recommendations – Chapter <124>  The USP EPO for bioassay RS is assigned a unitage which represents its activity in both in vivo assays and in vitro assays. – If the ratios of in vitro to in vivo potency for the material being tested and the USP EPO for bioassay RS are equivalent, then the USP EPO for bioassay RS can be used directly in the in vitro assay to calibrate the material being tested. – However, if these ratios are not equivalent, then the standard cannot be used with its assigned in vitro potency in the in vitro assay. – Instead, the ratio determined for the material being tested should then be used to assign a process-specific, in vitro assay unitage to the USP EPO for bioassay RS. The USP EPO for bioassay RS, with its adjusted in vitro assay unitage, can then be used in the in vitro assay to transfer the unitage from the USP EPO for bioassay RS to the material being tested.
  • 142. Modernizing Compendial Bioassay/Bioidentity Tests: Challenges  For small proteins or peptides: when can a bioassay be replaced by a physicochemical method? And, can the method stand alone or is bioidentity assurance needed?  Assays for biological products with more than one activity  Commutability of assays and reference materials  Availability of public methods and data: – Intellectual property barriers  Equivalence determination between assays: – Challenges in establishing equivalent results between different biological systems: animal vs. cell-based assays vs. physico-chemical, etc.

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